阅读说明：本技术 新黄酮类化合物Hip B和Hip C及其用途 (Novel flavonoid compounds Hip B and Hip C and application thereof ) 是由 王洪伦 铁芳芳 丁金 于 2020-06-29 设计创作，主要内容包括：本发明公开了新黄酮类化合物Hip B和Hip C及其用途,对沙棘籽粕浸膏通过MCI凝胶柱洗脱,再依次经C18色谱柱、二醇基色谱柱、C18色谱柱半制备分离后,得到本发明新黄酮类化合物Hip B和Hip C。本发明还提供了新化合物Hip B和Hip C在制备预防和/或治疗脂代谢紊乱的药物中的应用,通过对化合物Hip B和Hip C进行进行活性研究,验证了化合物Hip B和Hip C能够抑制细胞脂质生成,可用于制备预防或治疗肥胖症、高血脂等脂代谢紊乱疾病的药物。(The invention discloses novel flavonoid compounds Hip B and Hip C and application thereof, wherein sea buckthorn seed meal extractum is eluted by an MCI gel column, and then is subjected to semi-preparative separation by a C18 chromatographic column, a diol basic color chromatographic column and a C18 chromatographic column in sequence to obtain the novel flavonoid compounds Hip B and Hip C. The invention also provides application of the new compounds Hip B and Hip C in preparing medicaments for preventing and/or treating lipid metabolism disorder, and the activity research on the compounds Hip B and Hip C proves that the compounds Hip B and Hip C can inhibit cell lipid generation and can be used for preparing medicaments for preventing or treating lipid metabolism disorder diseases such as obesity, hyperlipidemia and the like.)

1. A flavonoid compound is characterized by having a structure shown in a formula I':

wherein R is₁、R₂、R₃、R₄Each independently selected from the group consisting of H, C1-3 alkyl.

2. Compound I' according to claim 1, characterized in that R₁、R₂、R₃、R₄Each independently selected from H, methyl; further, the compound I' is selected from the following compounds:

3. the use of a compound according to claim 1 or 2, and pharmaceutically acceptable salts, hydrates or solvates thereof, for the preparation of a product for the prevention and/or treatment of a disease associated with a disorder of lipid metabolism.

4. The use according to claim 3, wherein the product is a product with lipid-lowering effect;

further, the product is a product for reducing the content of cellular lipid droplets and/or triglycerides.

5. The use according to claim 4, wherein the product is a product inhibiting the production of cellular lipids.

6. The use according to claim 5, wherein said product is at least one of a FAS inhibitor, SREBP1 inhibitor, SCD-1 inhibitor.

7. The use according to any one of claims 1 to 6, wherein the product is at least one of a product for preventing and/or treating obesity, hypertension, hyperlipidemia, cardiovascular disease, and metabolic syndrome-related disease.

8. Use of a compound according to claim 1 or 2 for the preparation of at least one of a FAS inhibitor, a SREBP1 inhibitor, an SCD-1 inhibitor.

9. A product comprising a compound I' according to claim 1 or 2.

10. The product of claim 9, wherein the product comprises compound XVIII and/or compound XIX; further, the product also comprises a compound IV and/or a compound XVII, and the structural formulas of the compound IV and the compound XVII are as follows:

Technical Field

The invention relates to the field of flavonoid compounds and medical application thereof, in particular to novel flavonoid compounds Hip B and Hip C and application thereof.

Background

Seabuckthorn (the name of Latin, Hippophae rhamnoides Linn.) is a deciduous shrub of the genus Hippophae of the family Elaeagnaceae, and is drought-resistant and sand-resistant, and can survive in saline-alkali lands, thus being widely used for water and soil conservation. Sea buckthorn is planted in great amount in northwest China for desert greening.

The sea buckthorn is rich in nutrition, and contains active substances such as various vitamins, flavonoids, triterpenoids, oil and fatty acid, phenols, volatile oil, trace elements, phospholipids, 5-hydroxytryptamine and various amino acids and proteins required by human bodies.

Seabuckthorn seeds are commonly used for pressing oil to prepare seabuckthorn oil, which contains 206 active substances beneficial to human bodies, wherein 46 bioactive substances comprise a large amount of vitamin E, vitamin A, flavone and the like. A large amount of seabuckthorn seed meal is produced after oil extraction and is usually discarded as waste materials, or produced into dry powder to be used as feed, and the price is very low. If the utilization value of the seabuckthorn seed meal can be developed more, the method has great economic significance.

In addition, many active substances contained in the sea buckthorn are not separated and identified, and if the new chemical components and the pharmacological action of the new chemical components in the sea buckthorn can be studied more deeply and finely and the active mechanism can be studied, more safe and effective natural plant-derived medicines capable of treating diseases are expected to be developed.

Disclosure of Invention

The invention provides novel flavonoid compounds Hip B and Hip C extracted from seabuckthorn seed meal and application thereof in preparing medicaments for preventing and/or treating diseases related to lipid metabolism disorder, wherein the compounds have the effects of inhibiting generation and accumulation of cell lipid.

The invention provides a novel flavonoid compound in sea buckthorn, which has a structure shown in a formula I':

wherein R is₁、R₂、R₃、R₄Each independently selected from the group consisting of H, C1-3 alkyl. Further, R₁、R₂、R₃、R₄Each independently selected from H, methyl; further, the compound I' is selected from the following compounds:

the invention also provides the application of the compound I' and pharmaceutically acceptable salts, hydrates or solvates thereof in preparing products for preventing and/or treating diseases related to lipid metabolism disorder.

Further, the product is a product with lipid-lowering effect, such as lipid-lowering drugs and the like.

The product or the lipid-lowering medicine with the lipid-lowering effect is a product or a medicine for inhibiting the generation of lipid drops in fat cell cytoplasm and reducing the content of triglyceride.

Further, the product is a product for reducing the content of cellular lipid droplets and/or triglycerides.

Further, the product is a product for inhibiting the production of cellular lipids.

Further, the product is at least one of FAS inhibitor, SREBP1 inhibitor, and SCD-1 inhibitor.

Further, the product is at least one of products for preventing and/or treating obesity, hypertension, hyperlipidemia, cardiovascular diseases, and metabolic syndrome related diseases.

Obesity, hypertension, hyperlipidemia, cardiovascular diseases, metabolic syndrome, etc. are closely related to the content of plasma lipids such as Triglyceride (TG), Free Cholesterol (FC), Cholesterol Ester (CE) and phospholipids, and when plasma lipids are reduced to a certain concentration range in vivo, these diseases can be effectively controlled or treated. Experiments prove that the compound IV can effectively inhibit lipid droplet accumulation, reduce TG content, regulate the expression level of fatty acid synthesis related enzymes (such as SCD-1 and FAS) and important transcription factors (such as SREBP1) in adipogenesis gene regulation through related signal pathways, thereby inhibiting the lipogenesis of cells, and can be used for preparing products for preventing and/or treating obesity, hypertension, hyperlipidemia, cardiovascular diseases and metabolic syndrome related diseases.

The invention also provides application of the compound I' in preparing at least one of FAS inhibitor, SREBP1 inhibitor and SCD-1 inhibitor.

The FAS inhibitor, SREBP1 inhibitor and SCD-1 inhibitor are drugs for reducing protein expression levels of FAS, SREBP1 and SCD-1 in cells.

The invention also provides a product comprising compound I'.

Further, the product is at least one of a medicine, a health product and a food.

Experiments show that the compounds IV, XVII, XVIII and XIX in the embodiment of the invention can effectively inhibit lipid accumulation in cells, have the effects of repairing myocardial damage, protecting myocardial cells and protecting nerves, and can be used for developing products for preventing and/or treating related diseases.

The product comprises at least compound XVIII and/or compound XIX.

On this basis, the present invention also provides a composition comprising not only compound XVIII and/or compound XIX, but also compound IV and/or compound XVII.

Products prepared based on the combination of the compounds belong to the protection scope of the invention.

In the present invention, the percentages before the methanol aqueous solution and the ethanol aqueous solution are both expressed by volume fraction, for example, "40% methanol aqueous solution" is a methanol aqueous solution with a methanol volume fraction of 40%, and the same applies to the rest.

The invention has the beneficial effects that:

(1) the invention extracts new flavonoid compounds Hip B and Hip C from seabuckthorn seed meal and carries out activity research on the flavonoid compounds Hip B and Hip C, and the results show that the flavonoid compounds Hip B and Hip C can inhibit lipid drop accumulation, reduce TG content, and regulate the expression level of important transcription factors in regulation of fatty acid synthesis related enzyme and lipogenesis genes through related signal pathways, thereby inhibiting cell lipogenesis, and providing a powerful direction for researching and using the flavonoid compounds Hip B and Hip C for lipid metabolism disorder diseases such as obesity, cardiovascular diseases and the like.

(2) The invention more comprehensively exploits the medicinal value of the seabuckthorn, expands the clinical application of the seabuckthorn, provides more reference basis for developing potential plant-derived medicaments for treating diseases related to lipid metabolism disorder, realizes the waste utilization of seabuckthorn seed meal and changes waste into valuable.

Drawings

FIG. 1 is an analysis chromatogram of each component sample of seabuckthorn seed meal;

FIG. 2 is a one-dimensional preparation of a sample of the SH5 component of seabuckthorn seed meal;

FIG. 3 is a two-dimensional analysis of samples of the components in SH 5;

FIG. 4 is a purity analysis of peak 1-19 compounds in seabuckthorn seed meal;

FIG. 5 is of Compound Hip B¹H NMR spectrum;

FIG. 6 is of compound Hip B¹³C NMR spectrum;

FIG. 7 is an HSQC map of compound HipB;

FIG. 8 is a HMBC diagram of compound Hip B;

FIG. 9 is a HHCOSY plot of compound Hip B;

FIG. 10 is a NOESY plot of compound Hip B;

FIG. 11 is of compound Hip C¹H NMR spectrum;

FIG. 12 is of Compound Hip C¹³C NMR spectrum;

FIG. 13 is an HSQC map of compound Hip C;

FIG. 14 is a HMBC diagram of compound Hip C;

FIG. 15 is a HHCOSY plot of compound Hip C;

FIG. 16 is a NOESY plot of compound Hip C;

FIG. 17 is the effect of Oleic Acid (OA) on cell viability of HepG2 cells;

FIG. 18 is a graph of the effect of Oleic Acid (OA) on lipid accumulation in HepG2 cells;

FIG. 19 is the effect of Oleic Acid (OA) on FAS protein expression in HepG2 cells;

FIG. 20 is the effect of compounds Hip B, Hip C on HepG2 cell viability;

FIG. 21 is a graph of the effect of compounds Hip B, Hip C on lipid accumulation in HepG2 cells;

FIG. 22 is a graph of the effect of compounds Hip B, Hip C on adipogenic proteins of HepG2 cells;

fig. 23 is a morphology micrograph of DoX-induced H9c2 cardiomyocytes (100 ×, n ═ 3);

FIG. 24 is a graph of the effect of DoX on H9c2 cardiomyocyte viability (II)n＝3)；

FIG. 25 is a graph of DoX versus H9c2 cardiomyocyte culture supernatant NO content levels ((R))

n＝3)；

FIG. 26 is a graph of DoX vs H9c2 cardiomyocyte culture supernatant LDH content level (

n＝3)；

FIG. 27 shows the effect of Hip B and C compounds on the viability of H9C2 cardiomyocytes (

n＝3)；

FIG. 28 shows LDH levels induced by H9C2 myocardial cell injury by DoX pair of compounds Hip B and C (II)

n＝3)；

FIG. 29 shows NO levels of DoX-induced H9C2 myocardial cell injury by Hip B and C compounds

n＝3)；

FIG. 30 is a graph showing the effect of Hip B and C compounds on DoX-induced H9C2 apoptotic proteins in cardiomyocytesn＝3)；

FIG. 31 is a graph of the effect of 5. mu.M ATRA and 40nM OA on synaptic length of SH-SY5Y cells;

FIG. 32 is the effect of compounds Hip B, Hip C on SH-SY5Y cell viability;

FIG. 33 is a graph of the effect of compounds Hip B, Hip C on the morphology of AD cell models;

FIG. 34 shows the effect of hippophae rhamnoides neoflavonoids on tau phosphorylation in SH-SY5Y cells.

Detailed Description

The preparation and application of the novel flavonoid compound are further described by specific examples and specific experiments, and the effects of the novel flavonoid compound on inhibiting the accumulation of cell lipid and protecting myocardial cell injury and neuroprotection are verified and explained.

In the embodiment of the invention, the sea-buckthorn (Hippophae rhamnoides Linn.) seed meal material is the seed of sea-buckthorn produced in Datong county of Xining City of Qinghai province and utilizes supercritical CO₂And extracting seed meal after the seed oil is extracted.