Use of fatty acid compounds for the preparation of hypoglycemic agents

Abstract

This invention discloses the application of fatty acid compounds in the preparation of hypoglycemic drugs, belonging to the field of pharmaceutical technology. The fatty acid compounds are selected from any one or at least a combination of two of (5Z,9Z)-tetracosano-5,9-dienoic acid, gingerol A, and (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol. This invention discovers that the compounds can inhibit... α - The activity of glucosidase reduces postprandial hyperglycemia and can be used to prepare hypoglycemic drugs. This invention also provides the application of the aforementioned fatty acid compounds in the preparation of... α Applications of glucosidase inhibitors.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of fatty acid compounds in the preparation of hypoglycemic drugs. Background Technology

[0002] Diabetes mellitus is a complex metabolic disease characterized by hyperglycemia, mainly divided into type 1 and type 2 diabetes. With improved living standards, an aging population, and increasing obesity rates, diabetes continues to be prevalent globally. Persistent hyperglycemia and long-term metabolic disorders in diabetes can lead to damage and dysfunction of various organs and tissues throughout the body, particularly the cardiovascular, ocular, renal, and nervous systems, and are a high-risk factor for major diseases such as cardiovascular disease, cancer, and COVID-19. Currently, the main antidiabetic hypoglycemic drugs used clinically include insulin and metformin. α Drugs include glucosidase inhibitors (such as acarbose and voglibose), GLP-1 receptor agonists (such as liraglutide), DPP-4 inhibitors (such as sitagliptin), insulin secretagogues such as sulfonylureas (such as glimepiride), and insulin sensitizers (such as biguanides and thiazolidinediones). However, these drugs still have problems such as inconvenient administration, inability to stably control blood glucose levels, or significant side effects (such as liver and kidney toxicity, edema, and severe gastrointestinal reactions). Therefore, the development of novel hypoglycemic drugs or functional foods remains a research hotspot in the field of antidiabetic treatment.

[0003] α - Glucosidase belongs to the class of oligosaccharide hydrolases. Its inhibitors reduce the degradation of sugars and delay the digestion and absorption of sugars by competitively inhibiting the action of glycosidases on the villi of the small intestinal epithelium. This effectively reduces the peak postprandial blood glucose concentration in diabetic patients and achieves the goal of blood glucose control. α Glucosidase inhibitors (such as acarbose) are a class of oral hypoglycemic agents widely used in clinical practice, and can be widely used for blood glucose control in patients with type 1 and type 2 diabetes. However, acarbose and other similar drugs are currently the most widely used in clinical practice. α - Glucosidase inhibitors can easily cause gastrointestinal disturbances such as bloating, diarrhea, and abdominal pain, as well as liver damage. Therefore, the development of safer and more effective new... α - Glucosidase inhibitors are of great significance. Summary of the Invention

[0004] To address the problems of existing technologies, this invention provides a new use for fatty acid compounds in the preparation of hypoglycemic drugs. Specifically, this invention discovers that three fatty acid compounds can inhibit... α - Glucosidase activity reduces postprandial hyperglycemia and can be used to prepare hypoglycemic drugs or α - Glucosidase inhibitor.

[0005] To achieve this objective, the present invention adopts the following technical solution: In a first aspect, the present invention provides the use of fatty acid compounds or their pharmaceutically acceptable salts or esters in the preparation of hypoglycemic drugs, wherein the fatty acid compounds are selected from any one or at least a combination of two of formulas I to III:

[0006]

[0007] .

[0008] Formula I is called (5) in English. Z 9 Z Formula II is tetracosa-5,9-dienoic acid; Formula III is gingerglycolipid A; Formula IV is (2... S )-3- O -[ α -D-galactopyranosyl-(1→6)- β -D-galactopyranosyl]-1,2-di- O -[(9 Z 12 Z 15 Z )-octadeca-9,12,15-trienoyl]- sn -glycerol, also known as (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol. All of the above compounds can be isolated from the medicinal plant *Eclipta prostrata* and exhibit high safety.

[0009] Preferably, the drug is able to inhibit the intestines α - It can reduce postprandial hyperglycemia in patients with type 1 or type 2 diabetes by inhibiting glucosidase activity. Through experimental research, the inventors discovered that the fatty acid compounds have the effect of... α -IC50 glucosidase inhibitory activity 50 The values ​​were 2.42 ± 0.21, 12.42 ± 0.45, and 14.36 ± 0.72 μmol / L, respectively, indicating that they had an effect on... α -Glucosidase has good inhibitory activity. It can be used to prepare blood glucose-regulating drugs, especially those that lower postprandial hyperglycemia.

[0010] Preferably, the drug further comprises pharmaceutically acceptable excipients, including any one or a combination of at least two of the following: carrier, diluent, filler, binder, wetting agent, disintegrant, emulsifier, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH adjuster, antioxidant, antibacterial agent, or buffer. Preferably, the dosage form of the drug is tablets, capsules, injections, granules, or oral liquids; Preferably, the daily dosage of the fatty acid compound is 0.01 to 100 mg / kg based on body weight.

[0011] Furthermore, the fatty acid compounds of the present invention, or their pharmaceutically acceptable salts, esters, solvates, or prodrugs, can also be used in combination with other drugs to achieve better hypoglycemic effects.

[0012] In this invention, the route of administration of the drug can be selected from any one of oral administration, sublingual administration, intravenous injection, intramuscular injection or subcutaneous injection, depending on actual needs.

[0013] Secondly, the present invention provides the use of fatty acid compounds or their pharmaceutically acceptable salts or esters in the preparation of α-glucosidase inhibitors, wherein the fatty acid compounds are selected from any one or at least a combination of two of formulas I to III:

[0014]

[0015] .

[0016] Thirdly, the present invention provides the use of fatty acid compounds or pharmaceutically acceptable salts or esters thereof in the preparation of reagents for inhibiting α-glucosidase activity in vitro, wherein the fatty acid compounds are selected from any one or at least a combination of two of formulas I to III:

[0017]

[0018] .

[0019] This invention unexpectedly discovered that fatty acid compounds represented by formulas I to III ((5Z,9Z)-tetracosano-5,9-dienoic acid, gingerol A, (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol) can inhibit α-Glucosidase activity, which is related to α -IC50 glucosidase inhibitory activity 50 The concentrations were 2.42 ± 0.21, 12.42 ± 0.45, and 14.36 ± 0.72 μmol / L, respectively. These compounds are widely found in traditional Chinese medicinal herbs such as Eclipta prostrata, exhibit low toxicity and high safety. They can be used to prepare hypoglycemic drugs, especially for reducing postprandial hyperglycemia, and hold promise for development into a new generation of [hypoglycemic agents]. α - Glucosidase inhibitors are a class of hypoglycemic drugs. Attached Figure Description

[0020] Figure 1 For fatty acid compounds α - Results of the inhibition experiment of glucosidase, where a is (5Z,9Z)-tetracosano-5,9-dienoic acid, b is gingerol A, and c is (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol. Detailed Implementation The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0021] The processes, conditions, reagents, and experimental methods used in implementing this invention, except as mentioned below, are all common knowledge and general knowledge in the field, and this invention does not have any particular limitations. Experimental methods in the embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer.

[0022] Unless otherwise stated, all technical terms and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. However, in the event of any conflict, the specification containing the definitions shall prevail.

[0023] Eclipta prostrata was purchased from Yunnan Ke'en Pharmaceutical Slices Co., Ltd. α -Glucosidase, p-nitrophenyl- β -D-galactopyranoside and acarbose were purchased from Sigma.

[0024] Example 1: Preparation of fatty acid compounds 20 kg of dried Eclipta prostrata (a type of medicinal herb) was extracted three times with 90% industrial ethanol at room temperature, each extraction lasting three days. The extracts were combined and concentrated under reduced pressure to obtain 2.39 kg of Eclipta prostrata ethanol extract. The Eclipta prostrata ethanol extract was then dissolved in 15 times its volume of warm water and stirred until fully dissolved. It was then extracted six times each with petroleum ether and n-butanol. The petroleum ether, n-butanol, and water fractions were concentrated under reduced pressure to obtain the petroleum ether extract (660.7 g), n-butanol extract (289.6 g), and water fraction (1222.8 g), respectively. The fractions were then dried and stored at low temperature.

[0025] The petroleum ether extract obtained above was separated by silica gel column chromatography and eluted with a petroleum ether-ethyl acetate gradient (100:0~0:1) to obtain four fractions B1-B4; B.2 was subjected to repeated column chromatography on silica gel, Sephadex LH-20, and preparative HPLC to separate and purify (5Z,9Z)-tetracos-5,9-dienoic acid (38.5 mg, purity ≥98%).

[0026] The n-butanol extract obtained above was separated by silica gel column chromatography and eluted with a gradient of petroleum ether-ethyl acetate (1:0~0:1) and ethyl acetate-methanol (100:1~0:1) to obtain three components B5-B7. B5 was eluted with a gradient of petroleum ether / ethyl acetate (5:1~0:1) and purified by Sephadex LH-20, RP-C18 reversed-phase column chromatography, and semi-preparative HPLC to obtain gingerol A (13.7 mg, purity ≥98%) and (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol (11.2 mg, purity ≥98%).

[0027] The compound 1 H NMR, 13 Comparison of relevant data such as C NMR and MS with literature revealed that the structures of the three compounds were (5Z,9Z)-tetracos-5,9-dienoic acid, gingerol A, and (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol.

[0028] Its structural identification data are as follows: (5Z,9Z)-Ticosico-5,9-dienoic acid: colorless oil, molecular formula C2 24 H 44 O2, ESI-MS m / z 365 [M+H]+ . 1 H NMR (500 MHz, CDCl3) δ H 5.36 (4H, m, H-5, 9), 2.35 (6H, t, J = 7.5 Hz, H-2,6, 10), 2.02 (6H, m, H-4, 7, 8), 1.63 (2H, m, H-3), 1.33 (2H×13, m, H-11–23), 0.98 (3H, t, J = 7.5 Hz, H-24); 13 C NMR (125 MHz, CDCl3) δ C 180.4 (C-1), 130.4(C-5), 130.2 (C-5), 128.5 (C-9), 128.0 (C-10), 34.3 (C-2), 32.1 (C-22), 29.2–23.9 (C-12–21), 27.4 (C-4, 7, 8), 25.8 (C-11), 24.9 (C-3), 22.9 (C-23), 14.3(C-24). Ginger glycolipid A: Yellow amorphous powder, molecular formula C 33 H 56 O 14 ESI-MS m / z : 711 [M+Cl] - . 1 H NMR (500 MHz, CD3OD) δ H 5.85–5.25 (6H, m, H-9, 10, 12, 13, 15, 16), 4.91 (1H, s, H-1'''), 4.47 (1H, d, J = 11.6 Hz, H-1''), 4.28 (2H, m, H-3'), 2.85 (1H, dd, J =12.8, 7.1 Hz, H-2), 2.37 (4H, dd, J = 13.0, 6.7 Hz, H-11, 14), 2.13 (2H, m, H-8), 1.65 (4H, d, J= 4.9 Hz, H-3, 17), 1.37 (8H, m, H-4, 5, 6, 7), 1.03 (3H, m,H-18); 13 C NMR (125 MHz, CD3OD) δ C 174.8 (C-1), 132.8 (C-16), 131.1 (C-9), 129.2(C-13), 129.1 (C-12), 128.9 (C-10), 128.3 (C-15), 105.3 (C-1''), 100.6 (C-1'''), 74.7 (C-5''), 74.6 (C-3''), 72.5 (C-5'''), 72.4 (C-2''), 71.8 (C-1'),71.5 (C-3'''), 71.1 (C-4'''), 70.2 (C-2'''), 70.1 (C-4''), 68.8 (C-2'), 67.8(C-6''), 64.0 (C-3'), 62.8 (C-6'''), 34.9 (C-2), 30.7 (C-4), 30.3 (C-5, 7), 30.2 (C-6), 28.2 (C-8), 26.6 (C-14), 26.4 (C-11), 26.0 (C-3), 21.5 (C-17),14.7 (C-18). (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol: Yellow amorphous powder, molecular formula C 51 H 84 O 15 ESI-MS m / z 935 [MH] - . 1 H NMR (500 MHz, CD3OD) δ H 5.34 (12H, m, H-9', 10', 12', 13',15', 16', 9'', 10'', 12'', 13'', 15'', 16''), 5.24 (1H, m, H-2), 4.43 (1H,dd, J = 12.1, 2.8 Hz, H-1b), 4.23 (1H, dd, J= 6.9, 12.1 Hz, H-1a, ), 4.21 (1H,d, J = 6.7 Hz, H-1’’’’), 3.48 (2H, m, H-3), 2.80 (8H, t, J = 6.0 Hz, H-11’, 11’’,14’, 14’’), 2.31 (4H, d, J = 6.8 Hz, H-2’, 2’’), 2.07 (8H, dd, J = 14.6, 7.4 Hz,H-8’, 8’’, 17’, 17’’), 1.32 (16H, m, H-4’–7’, 4’’–7’’), 0.97 (6H, t, J = 7.5Hz, H-18’, 18’’); 13 C NMR (125 MHz, CD3OD) δ C 175.2 (C-1’), 174.9 (C-1’’), 132.9(C-16’, 16’’), 131.2 (C-15’, 15’’), 129.4 (C-12’, 12’’, 13’, 13’’), 129.1 (C-10’, 10’’), 128.4 (C-9’, 9’’), 105.5 (C-1’’’’), 100.8 (C-1’’’), 74.8 (C-3’’’’), 74.7 (C-5’’’’), 72.7 (C-2’’’’), 72.5 (C-5’’’), 71.9 (C-3’’’), 71.6(C-4’’’), 71.3 (C-2), 70.4 (C-2’’’), 70.2 (C-4’’’’), 68.9 (C-3), 67.9 (C-6’’’’), 64.2 (C-1), 63.0 (C-6’’’), 35.3 (C-2’), 35.1 (C-2’’), 30.9 (C-4’,4’’), 30.5 (C-5’, 5’’), 30.4 (6’, 6’’), 30.3 (C-7’, 7’’), 28.3 (C-8’, 8’’),26.7 (C-3’, 3’’), 26.5 (C-11’, 11’’), 26.2 (C-14’, 14’’), 21.7 (C-17’, 17’’),14.9 (C-18’, 18’’). Example 2 αEvaluation of glucosidase inhibitory activity Experimental principle: α - Glucosidase belongs to the class of oligosaccharide hydrolases. Its inhibitors reduce the degradation of sugars and delay the digestion and absorption of sugars by competitively inhibiting the action of glycosidases on the villi of the small intestinal epithelium. This effectively reduces the peak postprandial blood glucose concentration in diabetic patients and achieves the goal of blood glucose control. α - Glucosidase inhibitors (such as acarbose) have become a widely used class of oral hypoglycemic drugs in clinical practice. α - Screening for glucosidase inhibitor activity can be performed by comparing the enzyme with its substrate and nitrophenyl- β The in vitro enzymatic reaction of β-D-galactopyranoside (PNPG, a maltose analogue) was used for detection. α - After glucosidase is added to the substrate of the enzyme reaction, the substrate is catalyzed and decomposed into p-nitrophenol (PNP) and glucose. PNP is a colored substance with maximum absorption at around 400 nm, which can be measured by an enzyme-linked immunosorbent assay (ELISA) reader. The inhibitory activity of the sample can be calculated based on the OD value.

[0029] Experimental method: 10 μL of samples of different concentrations (three fatty acid compound samples prepared in Example 1) and 50 μL of... α - Glucosidase solution (0.1 U / mL) was added to each well of a 96-well plate containing PBS (pH 7.2) and incubated at 37°C for 15 minutes. Subsequently, 40 μL of PNPG (5 mmol / L) was added to each well and incubated at 37°C for 30 minutes. The reaction was terminated by adding 20 μL of Na₂CO₃ solution (0.5 mmol / L), and the OD value at 405 nm was measured using a Thermo Multiskan FC microplate reader. α The inhibition rate of glucosidase activity was calculated according to formula (1). α - Glucosidase inhibition rate.

[0030]

[0031] Among them, F a F represents the absorbance of the sample group. b This indicates the sample control group (using an equal volume of buffer solution instead). α -glucosidase solution) absorbance, F c F represents the absorbance of the control group (an equal volume of buffer solution instead of the sample solution). d This indicates the absorbance of the control / background group (an equal volume of buffer solution instead of the sample solution and enzyme solution).

[0032] See results Figure 1The results showed that (5Z,9Z)-tetracos-5,9-dienoic acid, gingerol A, and (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol had an effect on... α -IC50 glucosidase inhibitory activity 50 The values ​​were 2.42 ± 0.21, 12.42 ± 0.45, and 14.36 ± 0.72 μmol / L, respectively, showing significant differences. α - Glucosidase inhibitory activity.

[0033] Example 4 Tablet Preparation 1 g of compound (5Z,9Z)-tetracos-5,9-dienoic acid or gingerol A or (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol was mixed with 0.6 g of lactose and 0.5 g of starch. The mixture was moistened with water, sieved and dried, then sieved again. 0.1 g of magnesium stearate was added, mixed well, and compressed into tablets. 20 tablets were prepared, with a tablet weight of 110 mg and a content of 50 mg / tablet.

[0034] Example 5: Capsule Preparation 1 g of compound (5Z,9Z)-tetracosano-5,9-dienoic acid or gingerol A or (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol was mixed with 0.5 g of lactose and 0.1 g of magnesium stearate, sieved, and mixed evenly. The resulting mixture was then filled into 20 hard gelatin capsules, with a capsule weight of 80 mg and a content of 50 mg / capsule.

[0035] Example 6 Preparation of oral liquid preparation Add 1 g of compound (5Z,9Z)-tetracos-5,9-dienoic acid or gingerol A or (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol to the conventional additives used in the preparation of the oral liquid and purified water, and make up to 0.1 L. Under aseptic conditions, fill 20 ampoules, 5 ml per ampoule, content: 50 mg / ampoule.

[0036] Example 7 Granule Preparation 1 g of compound (5Z,9Z)-tetracos-5,9-dienoic acid or gingerol A or (2S)-3-O-[α-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl]-1,2-di-O-[(9Z,12Z,15Z)-octadecano-9,12,15-trienoyl]-sn-glycerol was mixed with 1 g of lactose and 0.6 g of starch. The mixture was moistened with water, sieved and dried, then sieved again. The mixture was then granulated and packaged into 20 bags, each weighing 130 mg, with a content of 50 mg / bag.

[0037] The above descriptions are merely several embodiments of this application and are not intended to limit this application in any way, i.e., they do not imply that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of raw materials for the product of the present invention, additions of auxiliary components, and selection of specific methods, all fall within the protection and disclosure scope of the present invention. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. The use of fatty acid compounds or their pharmaceutically acceptable salts or esters in the preparation of hypoglycemic drugs, wherein the fatty acid compounds are selected from any one or at least a combination of two of formulas I to III: 。 2. The application according to claim 1, characterized in that, The drug reduces postprandial hyperglycemia.

3. The application according to claim 2, characterized in that, The drug inhibits α - Decreased glucosidase activity leads to postprandial hyperglycemia.

4. The application according to any one of claims 1 to 3, characterized in that, The drug also includes one or more pharmaceutically acceptable excipients.

5. The application according to claim 4, characterized in that, The pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, diluents, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

6. The application according to any one of claims 1 to 3, characterized in that, The dosage form of the drug is capsules, granules, tablets, or oral liquid preparations.

7. The application according to any one of claims 1 to 3, characterized in that, The daily dosage of the fatty acid compounds is calculated based on body weight and ranges from 0.01 to 100 mg / kg.

8. The use of fatty acid compounds or their pharmaceutically acceptable salts or esters in the preparation of α-glucosidase inhibitors, wherein the fatty acid compounds are selected from any one or at least a combination of two of formulas I to III: 。 9. The use of fatty acid compounds or their pharmaceutically acceptable salts or esters in the preparation of reagents for inhibiting α-glucosidase activity in vitro, wherein the fatty acid compounds are selected from any one or at least a combination of two of formulas I to III: 。

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