阅读说明：本技术 一种atp钾离子通道促进剂组合物及其制备方法和应用 (ATP potassium ion channel promoter composition, and preparation method and application thereof ) 是由 不公告发明人 于 2020-11-13 设计创作，主要内容包括：本发明提出了一种ATP钾离子通道促进剂组合物,由以下原料制备而成：式Ⅰ的化合物盐及其生理学上可接受的其他盐、衍生物、溶剂化物、前药和立体异构体、钠钾ATP酶、瑞格列奈；所述式Ⅰ的化合物盐具有如式Ⅰ所示结构；其中,n＝1-5。该组合物具有在用抗精神病药治疗的受治疗者中预防或治疗体重增加、糖尿病或葡萄糖耐量减低的药物中的用途。(The invention provides an ATP potassium ion channel promoter composition, which is prepared from the following raw materials: salts of the compounds of formula (I) and their physiologically acceptable further salts, derivatives, solvates, prodrugs and stereoisomers, sodium potassium ATPase, repaglinide; the compound salt of the formula I has a structure shown in the formula I; wherein n is 1-5. The composition has use in a medicament for preventing or treating weight gain, diabetes, or impaired glucose tolerance in a subject treated with an antipsychotic agent.)

1. Salts of compounds of formula (I) and physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios:

wherein n is 1-5.

2. A compound salt of formula i as claimed in claim 1, which has one of the following structures:

3. the compound salts of formula i according to claim 1 and their physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios, characterized in that the chloride ion can be replaced by other anions, including acetate, formate, sulfate, phosphate, nitrate, bromide.

4. The compound salts of formula i according to claim 1 and their physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios, characterized in that the sodium ion can be replaced by other cations, including potassium, calcium, barium.

5. The compound salts of formula i according to claim 1 and their physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios, characterized in that they are prepared by the following process:

s1, mixing and reacting dimethyl amino alcohol, thionyl chloride and NaOH to generate an intermediate I with a structuren＝1-5；

S2, mixing the intermediate I, diazoxide and triethylamine for reaction to generate an intermediate II with the structure

And S3, adding dilute hydrochloric acid into the intermediate II, mixing and reacting, and then adding a dilute NaOH solution for neutralization to generate the compound salt of the formula I.

6. The compound salts of formula i according to claim 5 and their physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios, characterized in that the synthesis of the compound salts of formula i comprises in particular:

s1, dissolving dimethyl amino alcohol in dichloromethane, adding NaOH solid, placing in an ice bath, dropwise adding a dichloromethane solution of thionyl chloride while stirring, reacting for 1-2h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I;

s2, dissolving the intermediate I and triethylamine in acetonitrile, heating to 50-65 ℃, dropwise adding an acetonitrile solution of diazoxide, reacting for 3-5h, and filtering to generate an intermediate II;

s3, adding the intermediate II into a hot dilute hydrochloric acid solution, stirring and reacting for 10-30min, adding an excessive hot dilute NaOH solution, reacting for 10-20min, cooling, filtering, and washing the solid with distilled water to obtain a product, namely a compound salt of the formula I;

the mass ratio of the dimethyl amino alcohol, the thionyl chloride and the NaOH is 1 (2-3): (4-7);

the mass ratio of the intermediate I, diazoxide and triethylamine is 1: (1.1-1.3): (3-5);

the temperature of the hot dilute hydrochloric acid and the hot dilute NaOH solution is 80-90 ℃, the mass concentration of the dilute hydrochloric acid is 0.1-0.5mol/L, and the mass concentration of the dilute NaOH solution is 0.2-1 mol/L.

7. The ATP potassium ion channel promoter composition is characterized by being prepared from the following raw materials in parts by weight: 1 to 5 parts of compound salt of formula I and other physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, 0.2 to 0.5 part of sodium potassium ATPase and 1 to 3 parts of repaglinide.

8. The ATP potassium ion channel promoter composition as claimed in claim 3, which is prepared from the following raw materials in parts by weight: salts of the compounds of the formula I and their physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers 3 parts, sodium potassium ATPase 0.3 parts, repaglinide 2 parts.

9. A process for preparing a composition of a potassium ATP channel promoter according to any one of claims 1 to 8, comprising the steps of: dissolving compound salt of formula I and other physiologically acceptable salts, derivatives, solvates, prodrugs, stereoisomers and sodium potassium ATP enzyme thereof in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

10. Use of the ATP potassium channel promoter composition according to any one of claims 1-8 for the preparation of a medicament for the prevention or treatment of weight gain, diabetes or impaired glucose tolerance in a subject treated with an antipsychotic agent.

Technical Field

The invention relates to the technical field of biological pharmacy, in particular to an ATP potassium ion channel promoter composition, a preparation method and application thereof.

Background

ATP sensitive potassium (K)_ATP) Channels play an important role in a variety of tissues by coupling cellular metabolism to electrical activity. Has identified K_ATPThe channel is an octameric complex assembled at 4: 4 stoichiometry for two unrelated proteins: the first protein is the pore-forming subunit kir6.x, which forms the inward rectifying K + channel; the second is the ABC (ATP-binding cassette) transporter, also known as sulfonylurea receptor (SURX) (Babenko et al, An)nu.rev.physiol., 60: 667-687(1998)). The pore-forming subunit kir6.x is commonly used in many types of KATP channels and has two putative transmembrane regions (identified as TM1 and TM2) connected by a pore ring (H5). The subunit comprising the SUR receptor includes multiple transmembrane regions and two nucleotide binding folds.

According to their tissue location, K_ATPChannels exist in different subtypes or subclasses assembled from various combinations of SUR and Kir subunits. The combination of SUR1 and Kir6.x subunit (SUR1/Kir6.x) generally forms adipocytes and pancreatic B-cell type K_ATPChannels, whereas the SUR2A/Kir6.x and SUR2B/Kir6.x or Kir6.1 combinations typically form cardiac and smooth muscle type K, respectively_ATPChannels (Babenko et al, Annu. Rev. Physiol, 60: 667-687 (1998)). This also verifies that the channel can include a kir2.x subunit. Such potassium channels are inhibited by intracellular ATP and activated by intracellular nucleoside diphosphates. Such a K_ATPThe channels link the metabolic state of the cell to the plasma membrane potential and in this way play a major role in regulating cell activity. In most excitatory nerve cells, K_ATPThe channel is closed under normal physiological conditions and is open when tissue metabolism is reduced, for example when the (ATP: ADP) ratio is reduced. This promotes K + efflux and cellular hyperpolarization, thus preventing voltage-operated Ca²⁺Channel (VOC) open (prog. Res Research, (2001) 31: 77-80).

Potassium channel openers (PCO or KCO; also known as channel activators or channel antagonists) are a structurally distinct class of compounds that do not have a distinct common pharmacophore associated with their ability to antagonize the inhibition of KATP channels by intracellular nucleotides. Diazoxide is a K which stimulates in pancreatic beta cells_ATPPCO of channels (see Trube et al, Pflegers arc Eur J Physiol, 407, 493-99 (1986)). Pinadil and clolorkalim (chromakalim) are PCOs that activate cell membranes (see Escan et al, Biochem Biophys Res Commun, 154, 620 & 625 (1988); Babenko et al, J Biol Chem, 275(2), 717 & 720 (2000)). Reactivity to diazoxide has been shown to be located in the 6 th to 11 th predicted transmembrane region (TMD6-11) and first nucleotide binding fold of the SUR1 subunit.

Diazoxide is a compound having the formula 7-chloro-3-methyl-2H-1, 2, 4-benzothiadiazine 1, 1-dioxide (empirical formula C)₈H₇C_lN₂O₂S) are commercialized as three different formulations for the treatment of two different disease indications: (1) hypertensive emergency (2) hyperinsulinemic hypoglycemic conditions. Treatment of hypertensive emergencies Hyperstat IV, an aqueous formulation of diazoxide for intravenous use adjusted to pH 11.6 with sodium hydroxide. Hyperstat IV is administered as a bolus dose into the peripheral vein to treat malignant hypertension or sulfonylurea overdose. In this use, diazoxide acts to open potassium channels in vascular smooth muscle and to stabilize the membrane potential at resting levels, resulting in vascular smooth muscle relaxation.

The diazoxide synthesis methods reported worldwide are: in the united states patent 2986573, 5-chloro-2-nitrobenzenesulfonamide is used as a starting material and is reduced by iron powder to generate 5-chloro-2-aminobenzenesulfonamide; then carrying out cyclization reaction with triethyl orthoacetate to prepare the diazoxide. In the us patent 3345365, 2-aminobenzenesulfonamide is used as a starting material, and reacts with acetic anhydride to obtain 2-acetamido-N-acetylbenzenesulfonamide, then glacial acetic acid is used as a solvent, chlorine is introduced to perform chlorination to generate 5-chloro-2-acetamido-N-acetylbenzenesulfonamide, and diazoxide is obtained by solvent-free high-temperature cyclization and recrystallization refining. The above synthetic routes all have major disadvantages: the U.S. patent 2986573 uses iron powder for reduction, has high labor intensity and low efficiency, and also produces a large amount of iron mud which is difficult to treat, thereby causing environmental pollution; and triethyl orthoacetate is used in the cyclization step, so that the reaction yield is low, byproducts are more, the purification is difficult, and the product quality is not high. U.S. patent 3345365 uses acetic anhydride/pyridine as the acetylation agent, which produces large amounts of acetic acid-pyridine waste liquid that is difficult to recycle; the chlorination step adopts a direct chlorine chlorination mode, so that the environment is easily polluted, and the labor protection is not facilitated; in the cyclization step, solvent-free high-temperature cyclization is directly adopted, so that a large amount of impurities are generated, and the product purity is not high.

Therefore, the novel diazoxide compound is developed, and has the characteristics of convenient synthesis, wide raw material source, high yield, effective promotion of development of potassium channels, high biocompatibility and the like, thereby having remarkable practical significance.

Disclosure of Invention

The invention aims to provide an ATP potassium channel promoter composition, a preparation method and application thereof, and the ATP potassium channel promoter composition is convenient to synthesize, wide in raw material source, high in yield, capable of effectively promoting development of potassium channels and high in biocompatibility.

The technical scheme of the invention is realized as follows:

the present invention provides salts of the compounds of formula i and physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios:

formula I;

wherein n is 1-5.

As a further refinement of the present invention, the compound salt has one of the following structures:

as a further development of the invention, the chloride ion can be replaced by other anions, including acetate, formate, sulfate, phosphate, nitrate, bromide.

As a further improvement of the invention, the sodium ions may be replaced by other cations including potassium ions, calcium ions, barium ions.

As a further improvement of the invention, the compound salt of formula I is prepared by the following method:

s1, mixing and reacting dimethyl amino alcohol, thionyl chloride and NaOH to generate an intermediate I with a structuren＝1-5；

S2, mixing the intermediate I, diazoxide and triethylamine for reaction to generate an intermediate II with the structure

And S3, adding dilute hydrochloric acid into the intermediate II, mixing and reacting, and then adding a dilute NaOH solution for neutralization to generate the compound salt of the formula I.

As a further improvement of the invention, the synthesis method of the compound salt of the formula I specifically comprises the following steps:

s1, dissolving dimethyl amino alcohol in dichloromethane, adding NaOH solid, placing in an ice bath, dropwise adding a dichloromethane solution of thionyl chloride while stirring, reacting for 1-2h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I;

s2, dissolving the intermediate I and triethylamine in acetonitrile, heating to 50-65 ℃, dropwise adding an acetonitrile solution of diazoxide, reacting for 3-5h, and filtering to generate an intermediate II;

s3, adding the intermediate II into a hot dilute hydrochloric acid solution, stirring and reacting for 10-30min, adding an excessive hot dilute NaOH solution, reacting for 10-20min, cooling, filtering, and washing the solid with distilled water to obtain a product, namely a compound salt of the formula I;

the mass ratio of the dimethyl amino alcohol, the thionyl chloride and the NaOH is 1 (2-3): (4-7);

the mass ratio of the intermediate I, diazoxide and triethylamine is 1: (1.1-1.3): (3-5);

the temperature of the hot dilute hydrochloric acid and the hot dilute NaOH solution is 80-90 ℃, the mass concentration of the dilute hydrochloric acid is 0.1-0.5mol/L, and the mass concentration of the dilute NaOH solution is 0.2-1 mol/L.

The invention further provides an ATP potassium ion channel promoter composition, which is prepared from the following raw materials in parts by weight: 1 to 5 parts of compound salt of formula I and other physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, 0.2 to 0.5 part of sodium potassium ATPase and 1 to 3 parts of repaglinide.

As a further improvement of the invention, the health-care food is prepared from the following raw materials in parts by weight: salts of the compounds of the formula I and their physiologically acceptable other salts, derivatives, solvates, prodrugs and stereoisomers 3 parts, sodium potassium ATPase 0.3 parts, repaglinide 2 parts.

The invention further provides a method for preparing the ATP potassium ion channel promoter composition, which comprises the following steps: dissolving compound salt of formula I and other physiologically acceptable salts, derivatives, solvates, prodrugs, stereoisomers and sodium potassium ATP enzyme thereof in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

The present invention further protects the use of a composition of an ATP potassium channel promoter as described above for the preparation of a medicament for the prevention or treatment of weight gain, diabetes or impaired glucose tolerance in a subject treated with an antipsychotic agent.

The invention has the following beneficial effects: the novel diazoxide compound (compound salt of formula I) synthesized by the invention is a novel crystal salt component of diazoxide, has good water solubility and biocompatibility, simple synthesis method and high yield, and simultaneously, the sustained and controlled release pharmaceutical composition is prepared by adding gelatin, thereby increasing the utilization rate and the absorption rate of the drug;

the novel diazoxide compound and the repaglinide which is a hypoglycemic agent are compounded to form the effective ATP potassium ion channel promoter, the effect of the ATP potassium ion channel promoter has a synergistic effect, and the repaglinide which is a hypoglycemic agent promotes insulin secretion so as to reduce blood sugar content and regulate body weight. The sodium potassium ATPase can effectively promote ATP to be decomposed into ADP, thereby releasing energy, reducing blood sugar and controlling body weight.

The ATP potassium channel promoter composition of the present invention can be administered orally or parenterally, and the dosage varies depending on the drug, and is preferably 1 to 100mg per day for adults. For oral administration, the compound is first mixed with conventional pharmaceutical adjuvants such as excipient, disintegrant, binder, lubricant, antioxidant, coating agent, colorant, aromatic agent, surfactant, etc., and made into granules, capsules, tablets, etc.: for parenteral administration, the administration may be in the form of injection, infusion solution, suppository, or the like. In preparing the above formulation, conventional formulation techniques may be used.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.

FIG. 1 is a graph showing a comparison of glucose contents among groups of mice in test example 1 of the present invention;

FIG. 2 is a graph of mean drug-time curves of diazoxide in blood of various groups of rats in test example 2 of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

EXAMPLE 1 Synthesis of salts of Compounds of formula I

The synthetic route is as follows:

s1, dissolving 1mol of 2- (dimethylamino) ethanol (compound A) in 100mL of dichloromethane, adding 4mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2mol of thionyl chloride while stirring, reacting for 1h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 3mol of triethylamine in 100mL of acetonitrile, heating to 50 ℃, dropwise adding 20mL of acetonitrile solution of 1.1mol of diazoxide (compound C), reacting for 3h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 80 ℃ of 0.1mol/L, stirring for reaction for 10min, adding an excessive 0.2mol/L NaOH solution at 80 ℃, reacting for 10min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 95%.

Nuclear magnetic results: 1H NMR (600MHz, MeOD). delta.7.86 (s,1H), 7.52(d, 1H), 7.31(d, 1H), 3.26(t, 2H), 2.62(t, 2H), 2.27(s, 6H), 0.92(s, 3H). The nuclear magnetic detection results show that the structure is consistent with the predicted structure.

EXAMPLE 2 Synthesis of salts of Compounds of formula I

The synthetic route is as follows:

s1, dissolving 1mol of 6- (dimethylamino) -1-hexanol (compound A) in 100mL of dichloromethane, adding 7mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 3mol of thionyl chloride while stirring, reacting for 2h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 5mol of triethylamine in 100mL of acetonitrile, heating to 65 ℃, dropwise adding 20mL of acetonitrile solution of 1.3mol of diazoxide (compound C), reacting for 5h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 90 ℃ of 0.5mol/L, stirring for reaction for 30min, adding an excessive solution at 90 ℃ of 1mol/L NaOH, reacting for 20min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 85%.

Nuclear magnetic results: 1H NMR (600MHz, MeOD). delta.7.86 (s,1H), 7.52(d, 1H), 7.31(d, 1H), 3.26(t, 2H), 2.62(t, 2H), 2.55(t, 2H), 2.43(t, 2H), 2.35(t, 2H), 2.32(t, 2H), 2.27(s, 6H), 0.92(s, 3H). The nuclear magnetic detection results show that the structure is consistent with the predicted structure.

EXAMPLE 3 Synthesis of salts of Compounds of formula I

S1, dissolving 1mol of 2- (dimethylamino) ethanol (compound A) in 100mL of dichloromethane, adding 5mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.2mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 3.5mol of triethylamine in 100mL of acetonitrile, heating to 52 ℃, dropwise adding 20mL of acetonitrile solution of 1.15mol of diazoxide (compound C), reacting for 3.5h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 82 ℃ of 0.2mol/L, stirring for reaction for 15min, adding an excessive 0.4mol/L NaOH solution at 82 ℃, reacting for 12min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 90%.

EXAMPLE 4 Synthesis of salts of Compounds of formula I

S1, dissolving 1mol of 2- (dimethylamino) ethanol (compound A) in 100mL of dichloromethane, adding 6mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.8mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4.5mol of triethylamine in 100mL of acetonitrile, heating to 62 ℃, dropwise adding 20mL of acetonitrile solution of 1.25mol of diazoxide (compound C), reacting for 4.5h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a 0.4mol/L diluted bromic acid solution at 88 ℃, stirring for reaction for 25min, adding an excessive 0.8mol/L NaOH solution at 88 ℃, reacting for 18min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 94%.

Compound E has the following structure:

EXAMPLE 5 Synthesis of salt of Compound of formula I

The synthetic route is as follows:

s1, dissolving 1mol of 4- (dimethylamino) -1-butanol (compound A) in 100mL of dichloromethane, adding 5.5mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.5mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and reducing pressure to remove excessive thionyl chloride and dichloromethane to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4mol of triethylamine in 100mL of acetonitrile, heating to 58 ℃, dropwise adding 20mL of acetonitrile solution of 1.2mol of diazoxide (compound C), reacting for 4h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 85 ℃ of 0.3mol/L, stirring to react for 20min, adding an excessive 0.6mol/L NaOH solution at 85 ℃, reacting for 15min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 93%.

Nuclear magnetic results: 1H NMR (600MHz, MeOD). delta.7.86 (s,1H), 7.52(d, 1H), 7.32(d, 1H), 3.26(t, 2H), 2.62(t, 2H), 2.52(t, 2H), 2.44(t, 2H), 2.30(s, 6H), 0.91(s, 3H). The nuclear magnetic detection results show that the structure is consistent with the predicted structure.

EXAMPLE 6 Synthesis of salt of Compound of formula I

S1, dissolving 1mol of 2- (dimethylamino) ethanol (compound A) in 100mL of dichloromethane, adding 6mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.8mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4.5mol of triethylamine in 100mL of acetonitrile, heating to 62 ℃, dropwise adding 20mL of acetonitrile solution of 1.25mol of diazoxide (compound C), reacting for 4.5h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute nitric acid solution at 85 ℃ and 0.2mol/L, stirring for reaction for 25min, adding an excessive NaOH solution at 88 ℃ and 0.8mol/L, reacting for 18min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 94%.

Compound E has the following structure:

EXAMPLE 7 Synthesis of salt of Compound of formula I

S1, dissolving 1mol of 2- (dimethylamino) ethanol (compound A) in 100mL of dichloromethane, adding 6mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.8mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4.5mol of triethylamine in 100mL of acetonitrile, heating to 62 ℃, dropwise adding 20mL of acetonitrile solution of 1.25mol of diazoxide (compound C), reacting for 4.5h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute nitric acid solution at 85 ℃ of 0.2mol/L, stirring for reaction for 25min, adding an excessive KOH solution at 85 ℃ of 0.5mol/L, reacting for 18min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 94%.

Compound E has the following structure:

EXAMPLE 8 Synthesis of salt of Compound of formula I

S1, dissolving 1mol of 2- (dimethylamino) ethanol (compound A) in 100mL of dichloromethane, adding 6mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.8mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and removing excessive thionyl chloride and dichloromethane under reduced pressure to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4.5mol of triethylamine in 100mL of acetonitrile, heating to 62 ℃, dropwise adding 20mL of acetonitrile solution of 1.25mol of diazoxide (compound C), reacting for 4.5h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 85 ℃ and 0.5mol/L, stirring for reaction for 25min, and adding excessive Ba (OH) at 85 ℃ and 0.5mol/L₂The solution was reacted for 18min, cooled, filtered and the solid washed with distilled water to give the product, compound salt of formula i (compound E), in 94% overall yield.

Compound E has the following structure:

EXAMPLE 9 Synthesis of salt of Compound of formula I

The synthetic route is as follows:

s1, dissolving 1mol of 5- (dimethylamino) -1-pentanol (compound A) in 100mL of dichloromethane, adding 5.5mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of dichloromethane solution of 2.5mol of thionyl chloride while stirring, reacting for 1.5h, filtering, and reducing pressure to remove excessive thionyl chloride and dichloromethane to generate an intermediate I (compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4mol of triethylamine in 100mL of acetonitrile, heating to 58 ℃, dropwise adding 20mL of acetonitrile solution of 1.2mol of diazoxide (compound C), reacting for 4h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 85 ℃ of 0.3mol/L, stirring to react for 20min, adding an excessive 0.6mol/L NaOH solution at 85 ℃, reacting for 15min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 93%.

Nuclear magnetic results: 1H NMR (600MHz, MeOD). delta.7.87 (s,1H), 7.52(d, 1H), 7.32(d, 1H), 3.26(t, 2H), 2.60(t, 2H), 2.51(t, 2H),2.47(t, 2H), 2.39(t, 2H), 2.32(s, 6H), 0.92(s, 3H). The nuclear magnetic detection results show that the structure is consistent with the predicted structure.

EXAMPLE 10 Synthesis of salt of Compound of formula I

The synthetic route is as follows:

s1, dissolving 1mol of 3- (dimethylamino) -1-propanol (a compound A) in 100mL of dichloromethane, adding 5.5mol of NaOH solid, placing in an ice bath, dropwise adding 20mL of 2.5mol of dichloromethane solution of thionyl chloride while stirring, reacting for 1.5h, filtering, and reducing pressure to remove excessive thionyl chloride and dichloromethane to generate an intermediate I (a compound B);

s2, dissolving 1mol of the intermediate I (compound B) and 4mol of triethylamine in 100mL of acetonitrile, heating to 58 ℃, dropwise adding 20mL of acetonitrile solution of 1.2mol of diazoxide (compound C), reacting for 4h, and filtering to generate an intermediate II (compound D);

s3, adding the intermediate II (compound D) into a dilute hydrochloric acid solution at 85 ℃ of 0.3mol/L, stirring to react for 20min, adding an excessive 0.6mol/L NaOH solution at 85 ℃, reacting for 15min, cooling, filtering, and washing the solid with distilled water to obtain a compound salt (compound E) of the product formula I, wherein the total yield is 93%.

Nuclear magnetic results: 1H NMR (600MHz, MeOD). delta.7.86 (s,1H), 7.52(d, 1H), 7.31(d, 1H), 3.26(t, 2H), 2.60(t, 2H), 2.49(t, 2H), 0.95(s, 3H). The nuclear magnetic detection results show that the structure is consistent with the predicted structure.

EXAMPLE 11 ATP Potassium channel enhancer composition

The raw materials comprise the following components in parts by weight: 1 part of the novel diazoxide compound prepared in example 2, 0.2 part of sodium potassium ATPase, and 1 part of repaglinide.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

EXAMPLE 12 ATP Potassium channel promoter composition

The raw materials comprise the following components in parts by weight: 5 parts of the novel diazoxide compound prepared in example 4, 0.5 part of sodium potassium ATPase and 3 parts of repaglinide.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

EXAMPLE 13ATP Potassium channel enhancer composition

The raw materials comprise the following components in parts by weight: 2 parts of the novel diazoxide compound prepared in example 5, 0.2 part of sodium potassium ATPase and 1.2 parts of repaglinide.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

EXAMPLE 14ATP Potassium channel enhancer composition

The raw materials comprise the following components in parts by weight: 4 parts of the novel diazoxide compound prepared in example 6, 0.4 part of sodium potassium ATPase and 2.5 parts of repaglinide.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

Example 15ATP Potassium ion channel enhancer composition

The raw materials comprise the following components in parts by weight: 3 parts of the novel diazoxide compound prepared in example 7, 0.3 part of sodium potassium ATPase, and 2 parts of repaglinide.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

EXAMPLE 16ATP Potassium channel enhancer composition

The raw materials comprise the following components in parts by weight: 3 parts of the novel diazoxide compound prepared in example 8, 0.3 part of sodium potassium ATPase, and 2 parts of repaglinide.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

Comparative example 1

The novel diazoxide compound prepared in example 8 was not added, and other conditions were not changed, as compared with example 16.

The raw materials comprise the following components in parts by weight: 0.3 part of sodium potassium ATP enzyme and 5 parts of repaglinide.

The preparation method comprises the following steps:

dissolving sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

Comparative example 2

Compared with example 16, repaglinide was not added, and other conditions were not changed.

The raw materials comprise the following components in parts by weight: 5 parts of the novel diazoxide compound prepared in example 8 and 0.3 part of sodium potassium ATPase.

The preparation method comprises the following steps:

dissolving the novel diazoxide compound and the sodium-potassium ATP enzyme in deionized water, stirring and mixing uniformly, freezing and drying, and crushing the obtained solid to prepare the ATP potassium ion channel promoter composition.

Comparative example 3

In contrast to example 16, the novel diazoxide compound prepared in example 8 was replaced by commercially available diazoxide (supplied by Proglycem)

The raw materials comprise the following components in parts by weight: 3 parts of diazoxide, 0.3 part of sodium potassium ATP enzyme and 2 parts of repaglinide.

The preparation method comprises the following steps:

dissolving diazoxide and sodium potassium ATP enzyme in deionized water, stirring and mixing uniformly, adding repaglinide, stirring and mixing uniformly, freeze-drying, and crushing the obtained solid to obtain the ATP potassium ion channel promoter composition.

Test example 1 in vivo obesity test

Male 4-week-old B6 mice were housed in temperature-controlled (22 ℃) rooms with 12-hour light and 12-hour dark alternating periods, 5 mice per cage. The High Fat (HF) and Low Fat (LF) test diets contained 58% and 11% fat calories, respectively. One group of mice was fed HF diet at the first 4 weeks of the study; the remaining 45 mice were fed LF diet. Mice assigned to diet with LF maintained this diet throughout the study as a reference group for lean control mice. At the fourth week, all HF fed mice were subdivided into 9 groups of mice. The first group maintained the HF diet throughout the study as an obese control group. The remaining 8 groups of mice were fed an HF diet and concurrently administered the ATP potassium channel promoter composition prepared in examples 11-16, comparative examples 1-2, as a single dose administered by oral gavage at about 150mg of active ingredient per kg per day. Those animals were weighed weekly and food consumption in each cage was measured twice weekly until week 4 dietary changes, thus body weight and food intake were measured daily. Feeding efficiency (grams of body weight gained per calorie consumed) was calculated as the amount per cage. Samples for analysis of insulin, glucose and leptin were collected on day 24 (day 4 before diet change), day 32 (day 4 after diet change) and every two weeks thereafter. In all cases, the food was removed for 8 hours before the samples were collected. Glucose was analyzed by the glucose oxidase method and insulin and leptin concentrations were measured by the diabody RIA. The insulin assay is based on rat standards, whereas the leptin assay uses mouse standards. At the end of the study, a postprandial plasma sample was collected and analyzed for triglyceride and unesterified fatty acid concentrations. A subset of 10 animals in each group was sacrificed 4 weeks after drug treatment. White adipose tissue of the Epididymis (EWAT), Retroperitoneal (RP) fat, Interscapular Brown Adipose Tissue (IBAT) and gastrocnemius muscle were removed, trimmed and weighed. Percent body weight fat was estimated from the weight of epididymal fat pads. A subgroup of 5 animals per group was injected i.p. with 0.5g/kg glucose. 30 minutes after injection, plasma samples were collected and analyzed for glucose content by the glucose oxidase method. The results are shown in Table 1 and FIG. 1.

TABLE 1

Group of	Percent body weight fat (%)
		Example 11	24.5±2.6**##
Example 12	27.9±3.0**##
		Example 13	26.2±2.9**##
Example 14	25.5±3.1**##
		Example 15	24.5±3.4**##
Example 16	25.6±4.5**##
		Comparative example 1	44.5±3.4
Comparative example 2	40.6±4.5*#
		Model set	45.2±4.5**
Blank group	22.4±2.12

Note that: p <0.05 compared to blank group, # P <0.01, # P <0.05 compared to model group, # P < 0.01.

The ATP potassium channel promoter composition prepared by the invention has the effect of remarkably reducing the percentage body weight fat of obese mice, is beneficial to restoring blood sugar, and can be used in medicaments for preventing or treating weight gain, diabetes or impaired glucose tolerance.

The effects of the comparative example 1 and the comparative example 2 on the aspects of mouse weight regulation, blood sugar recovery and the like are obviously reduced without adding the novel diazoxide compound or repaglinide, wherein the comparative example 1 has no significance compared with a blank group and a model group. Therefore, the addition of the novel diazoxide compound or repaglinide has a synergistic effect.

Test example 2 pharmacokinetics

1. Reagent

Diazoxide control, available from the national institute of pharmacy, western medicine, university of Sichuan, lot number 20180205; the ATP potassium channel promoter compositions prepared in example 16 and comparative example 3; diazoxide, supplied by Proglycem, 50 mg/mL.

2. Material

Healthy rats 18 (provided by the animal center of the basic medical college of Sichuan university), weighing 190-.

3. Animal test model and selection of administration mode

An ideal animal model would be a pharmacokinetic study after gastric gavage in rats. The experiment adopts rat intragastric administration, aims to investigate the blood parameters of the drug in the rat body, and further studies whether the metabolic processes of the liquid composition and the Heptadine oral solution in the rat body have significant difference. The test adopts the mode of intragastric administration. Generally, the intragastric volume of the rat is about 2mL, and according to the literature report and the pre-test result, better detection sensitivity and analysis result can be obtained when the dosage is about 1.06 mg/kg. In this calculation, a rat of about 200g is administered in an amount of about 0.212 mg.

4. Test method

The rats are adaptively fed for 4 days before the test, and drinking water is not forbidden in the whole process; weighing body weight, calculating according to 1.06mg/kg, respectively taking blood from the wound surface of the self-broken tail after 30min, 45min, 1h, 2h, 3h, 4h, 6h, 8h, 12h and 24h after gastric lavage administration, and taking plasma after blood sample is anticoagulated by heparin sodium. Storing in a refrigerator at-24 deg.C.

5. Measurement of blood concentration

Plasma sample determination the collected plasma sample was taken and subjected to liquid chromatography detection with a control diazoxide solution (10. mu.g/mL), the position of the diazoxide absorption peak was determined, the diazoxide content in the plasma sample was obtained from the area of the absorption peak compared to the control, the diazoxide chronometric blood concentration data of four groups of rats are shown in Table 2, and the mean drug-time curves are shown in Table 2, respectively.

TABLE 2

From the mean plasma concentration-time curve (fig. 2), it is understood that, after the ATP potassium channel enhancer composition of example 16 was administered via gavage, the peak time of diazoxide was about 45min, a higher plasma concentration was achieved at about 30 minutes, and diazoxide in plasma was almost completely eliminated after 6 hours, and thus it is understood that the ATP potassium channel enhancer composition prepared according to example 16 of the present invention was rapidly absorbed, had a short peak time, and had a high effective dose after the administration via gavage. Is clearly superior to the conventional diazoxide tablet and comparative example 3.

Test example 3 treatment of obesity in human

The efficacy of ATP potassium channel promoter compositions prepared as described herein, example 16, comparative examples 1-3 and conventional diazoxide (supplied by Proglycom) can be tested in obese humans, as described by Alemzadeh (Alemzadeh et al, J Clin Endocr Metab, 83: 1911-. The subject is selected from a Body Mass Index (BMI) of greater than or equal to 30kg/m²And a moderately to morbidly obese adult. Each subject was subjected to a complete physical examination at the beginning of the evaluation, and body weight was measured on a standard electronic scale and body composition was measured by DEXA.

All subjects received a low calorie diet during the 1 week induction period prior to study initiation. This protocol was designed to exclude patients who could not be accommodated and to ensure stable body weight before treatment. Up to 75 subjects were tested at each dose of drug. Daily doses were set at 100, 200 and 300 mg/day. The daily dose was divided into 2 doses for administration. Doses of 50mg capsules or tablets were administered at one, two and three times per administration. The subjects took the drug daily for up to 12 months. Subjects were checked, weighed weekly, and asked for any side effects or concomitant illness.

A 24 hour dietary review was obtained from each subject. The dietary review was analyzed using standard computer software programs. All subjects received a low calorie diet, encouraging them to participate in regular exercise. Before the study began and after completion, the following laboratory tests were obtained: fasting glucose, fasting insulin, NEFA, and serum sodium (Na), potassium (K), and creatinine levels.

Insulin concentration was detected by RIA using a double antibody kit. Cholesterol and triglyceride concentrations were measured enzymatically. Plasma NEFA was detected by the enzymatic colorimetric method. Urine was collected over the corresponding 24 hours, which was used to measure total nitrogen measurement and detection substrate usage before and after the study.

The results are shown in Table 3.

Table 3 subjects with fasting glucose, fasting insulin, non-esterified fatty acids (NEFA), serum sodium (Na), potassium (K) and creatinine (n ═ 15)

All other clinical laboratory tests (including hematology, clinical serum chemistry, and urine chemistry) are within the normal range for obese subjects.

Compared with the prior art, the novel diazoxide compound (the compound salt of the formula I) synthesized by the invention is a novel crystal salt component of diazoxide, has good water solubility and biocompatibility, is simple in synthesis method and high in yield, and is prepared into a sustained and controlled release pharmaceutical composition by adding gelatin, so that the utilization rate and the absorption rate of the drug are increased;

the novel diazoxide compound and the repaglinide which is a hypoglycemic agent are compounded to form the effective ATP potassium ion channel promoter, the effect of the ATP potassium ion channel promoter has a synergistic effect, and the repaglinide which is a hypoglycemic agent promotes insulin secretion so as to reduce blood sugar content and regulate body weight. The sodium potassium ATPase can effectively promote ATP to be decomposed into ADP, thereby releasing energy, reducing blood sugar and controlling body weight.

The ATP potassium channel promoter composition of the present invention can be administered orally or parenterally, and the dosage varies depending on the drug, and is preferably 1 to 100mg per day for adults. For oral administration, the compound is first mixed with conventional pharmaceutical adjuvants such as excipient, disintegrant, binder, lubricant, antioxidant, coating agent, colorant, aromatic agent, surfactant, etc., and made into granules, capsules, tablets, etc.: for parenteral administration, the administration may be in the form of injection, infusion solution, suppository, or the like. In preparing the above formulation, conventional formulation techniques may be used.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.