CN104672144A - GPR119 agonist containing cyclopropyl hydrazide and nitrobenzene structures as well as preparation method and application thereof - Google Patents

Abstract

The invention relates to the field of medicines related to type II diabetes mellitus and in particular relates to a GPR119 agonist containing cyclopropyl hydrazide and nitrobenzene structures, a preparation method of the GPR119 agonist and an application of the GPR119 agonist to preparation of medicines related to type II diabetes mellitus. A structure of the GPR119 agonist is shown in a formula I in the specification.

Description

A kind of cyclopropyl hydrazides and nitrobenzene GPR119 agonist, preparation method and its usage

Technical field

The present invention relates to the pharmaceutical field of the treatment of type ii diabetes.More particularly, the present invention relates to the medicative a kind of GPR119 agonist containing cyclopropyl hydrazides and nitrobenzene structure of type ii diabetes, its preparation method and the purposes in pharmacy.

Background technology

Diabetes day by day seriously threaten the health of the mankind.At nowadays American, nearly 1,600 ten thousand people suffer from the misery that diabetes are brought.One patients with type Ⅰ DM is also referred to as insulin-dependent diabetes mellitus, belongs to autoimmune disorder.It causes because the pancreatic islet p-cells that can produce Regular Insulin is destroyed by self immune system, and at present in the world to this sick not cure method, therefore patient must accept injection of insulin treatment.If not insulin injection, cell cannot absorb glucose and obtain energy.The Childhood that the related symptoms of one patients with type Ⅰ DM coming across usually and adolescence.Because the course of disease is usually comparatively anxious, illness is obvious, and patient can be impelled initiatively to seek the help of medical procedure.Type-II diabetes is also referred to as non insulin dependent diabetes, shows as patient and lacks enough capability control own blood glucose levels.Type-II diabetes its caused by hypoinsulinism or insulin resistant (referring to that bodily tissue can not respond to the endocrine Regular Insulin of body rightly), that is type-II diabetes patient or be that the Regular Insulin of self secretion is insufficient, or is the Regular Insulin that can not effectively use self to secrete.Several factors can cause appearance and the development of insulin resistant, comprises heredity, obesity, advanced age and long term hyperglycemia etc.Although type-II diabetes may appear at all age group, generally occur in it grownup, so it is also called maturity-onset diabetes sometimes.But it should be noted that the sickness rate of type-II diabetes is in recent years soaring in children population.

With it diabetic subject, in blood and urine, the content of glucose raises, and causes diuresis, thirsty, hungry and the series of problems such as fat and protein metabolism.If not diagnosis and treatment in addition, diabetes can cause blind, gangrenous, and even the various life-threatening complication such as renal failure and heart trouble.

Type-II diabetes patient accounts for greatly the 90%-95% of diabetic subject's sum.In current Western society, about there is the grownup of 6% to suffer from type-II diabetes, cause 193 every year in the U.S., the death of 000 people, in all causes of death, occupy the 7th.Worldwide, be subject to the puzzlement of type ii diabetes more than 1.5 hundred million people, and this numeral is estimated to be doubled in 2025.Although some people easily suffers from diabetes, riseing mainly by mode of life, the full diet of sitting of current case because of the factor of heredity, and in developed country, general obesity caused.The type-II diabetes patient of general 80% is significantly overweight.The youngster suffering from now this disease is just increasing.Current type-II diabetes has been acknowledged as in 21 century in the world to one of significant threat of human health.

At present, people's treatment plan of having degree different to type-II diabetes.The most basic scheme is the combination of diet and exercise, also can coordinate pharmacological agent on this basis.The medicine used of current treatment diabetes, except Regular Insulin, also have following several specifically: Insulin secretagogues, such as sulfonylureas, it can improve the amount of pancreas beta cell excreting insulin; Hypoglycemic agents, such as metformin, it can reduce the amount of liver output glucose; Peroxisome proliferator-activated receptors-γ (PPAR-γ) agonist, such as glitazone drugs, it can strengthen the effect of Regular Insulin; Also have alpha-glucosidase inhibitor, it can hinder the output of glucose in intestines.But existing medicine also has the aspect of some shortcomings, comprise hypoglycemic side effect, body weight increases, the appearance of resistance, gastrointestinal problems, and oedema.The type-II diabetes needs of patients oral drug therapy of general 49%, the needs of patients insulin injection of general 40% also may use oral pharmaceutical simultaneously, and then the patient of general 10% can only use diet and exercise to carry out symptom management.

In order to new more effective therapy being introduced to the market, the research in several field is had to carry out at present.Its direction mainly comprises: the excessive production reducing glycogen, strengthen Regular Insulin absorbs path from glucose signals to cell transmission, increase the insulin secretion of the liver beta cell promoted by glucose, and try hard to solve fat and adjoint metabolism of fat and accumulation aspect problem.

GPR119 is a special target spot, and it is a member in g protein coupled receptor in rhodopsin family.Except being referred to as " GPR119 ", it also has other to identify, and includes but not limited to RUP3, Snorf25,19AJ, AXOR 20 and PS1.GPR119 is mainly expressed in beta Cell of islet in pancreatic tissue and PP cell, and enteron aisle L cell (secretion GLP-1) and K cell [secreting glucose-dependent-insulinotropic polypeptide (GIP)].Scientific experiment is verified, and exciting GPR119 can improve intracellular loops adenosine phosphate (cAMP) concentration, the stimulus-secretion coupling in activated cell, thus increases the GLP-1 of glucose dependency and the secretion of Regular Insulin.See T.Soga et al., Biochemical and Biophysical Research Communications, 2005,326,744-751, the inside some have document about GPR119.Also have Science Report recently, GPR119 agonist can reduce the apoptosis of people's enteron aisle L cell.

Type-II diabetes patient with it in, although the secretory volume of GLP-1 decreases, GLP-1 still keeps for the activity of beta cell, therefore has recently much for the research of GLP-1.These researchs indicate GLP-1 except stimulating body glucose dependency ground excreting insulin, also has other hypoglycemic mechanism, this includes but not limited to: the secretion suppressing glucagon after the meal, reduce and absorb nutrition to the speed in blood, and help control body weight by reducing appetite.Result of study shows, the therapy improving GLP-1 secretory volume can be applicable to various symptom and imbalance, include but not limited to metabolism disorder, gastrointestinal disturbance, inflammation, mental illness, depression, and neuropsychiatric disease includes but not limited to diabetes (type and two types), metabolic syndrome, obesity, poor appetite/excessively prosperous, becomes thin, anxiety, irritability, myocardial ischemia/reperfusion injury, senile dementia, and the disease of other central nervous systems.

But because GLP-1 can promptly be degraded by proteolytic enzyme DPP-IV, the application of exogenous GLP-1 on clinical treatment is very limited.According to reported in literature, have the analogue of several GLP-1 being used for treating type-II diabetes to be in the development phase, it is all through the polypeptide of modification, has the longer transformation period and activity is similar than the GLP-1 of people self secretion.With BYETTA be wherein trade(brand)name sell medicine be in this kind new medicine first by FDA approval listing.But these analogues need to be used by injection, this certain medicine that can increase GLP-1 secretion oral not as good as is more satisfactory.Truly have oral DPP-IV inhibitor on the market, it can reduce the degraded of GLP-1 thus improve GLP-1 level, such as is the sitagliptin that trade(brand)name renders to market with JANUVIA.If but there is a medicine can act on L-cell and beta cell simultaneously, promotes the endogenous excretion of GLP-1 and Regular Insulin, more benefits are had to the treatment of type-II diabetes, more promising.

Present invention finds the agonist of a kind of GPR119, it is by improving the level of GIP, GLP-1 and Regular Insulin, thus improves body to a certain extent to the processing power of glucose.Moreover, research shows GPR119 agonist, such as, molecule in the present invention, can promote to non-glucose dependency the secretion of incretin.Polypeptide GIP and GLP-1 is incretin, in the past 20 years, has a large amount of paper report GIP and GLP-1 to have diversified physiological action.Such as see Perry, T.et al., Curr.Alzheimer Res., 2005,377-385.After human body takes in nutritive substance, enteroendocrine cell K and L cell can secrete GIP and GLP-1 respectively.Although the mechanism that control GLP-1 secretes it be unclear that, the nerve conduction of the hormonal stimulation that perhaps the rapid rising of GLP-1 level can participate in owing to GIP in the dining rear short period of time, and after for some time, perhaps the lasting rising of GLP-1 level is caused by the direct activation of the nutritive substance in distal small intestine and colon to L-cell.GIP and GLP-1 is potent promotor, can strengthen the reaction that health is made the blood sugar raised, and improves the secretion of Regular Insulin.But patient demonstrates with it at type-II diabetes, although the effect of GLP-1 insulin secretion accelerating still keeps, the promoting insulin secretion of GIP declines.Puzzling is, type-II diabetes patient still has good reaction to the GIP that volume is injected, just lose susceptibility (Meier et al. to continuing a small amount of mode injected, Diabetes, 2004,53, S220-S224), so the exact cause of GIP activity decrease it be not immediately clear.Nearest research also shows, continues to use the derivative of fatty acid 14 days of a kind of long-acting GIP, contribute to the homeostasis (Irwin N.et al.J.Med.Chem., 49,1047-1054) that it maintains glucose to ob/ob mouse.

As can be seen here, GPR119 agonist has the value being applied in treatment diabetes and being associated in symptom, especially for type-II diabetes, fat, glucose intolerance, insulin resistant, Metabolic syndrome X, hyperlipidemia, hypercholesterolemia, and arteriosclerosis.

The invention discloses a kind of GPR119 agonist containing cyclopropyl hydrazides and nitrobenzene structure, these compounds can be used for the medicine preparing treatment type ii diabetes.

Summary of the invention

An object of the present invention is to provide a kind of GPR119 agonist with the excellent activity of formula I.

Another object of the present invention is to provide the method that preparation has the compound of formula I.

The compound that another object of the present invention is to provide containing formula I is treating the application in type ii diabetes as effective constituent.

Now in conjunction with object of the present invention, content of the present invention is specifically described.

The compound that the present invention has formula (I) has following structural formula:

Formula of the present invention (I) compound is synthesized by following route:

Compound II per and compound III, in generation substitution reaction, generate compound IV; Compound IV and hydrazine hydrate react, and generate compound V; Compound V and compound VI are reacted, and generate I.

Formula I of the present invention has the agonism of GPR119, can be used as the medicine of effective constituent for the preparation of type ii diabetes.The activity of formula I of the present invention is verified by receptor binding assays.

Formula I of the present invention is effective in quite wide dosage range.The dosage that such as every day takes, within the scope of 1mg-1000mg/ people, is divided into once or administration for several times.The actual dosage taking formula I can be decided according to relevant situation by doctor.

Embodiment

Below in conjunction with embodiment, the present invention is further illustrated.It should be noted that, following embodiment be only for illustration of, and not for limiting the present invention.The various changes that those skilled in the art's training centre according to the present invention is made all should within the protection domain required by the application's claim.

the synthesis of embodiment 1 Compound I-1

A. the synthesis of compound IV-1

Compound II per-1 (1.08g, 10mmol) with compound III (1.95g, 10mmol) be dissolved in the DMF of 20mL drying, stirred at ambient temperature, add solid carbonic acid potassium (4.15g, 30mmol) with 0.5g KI, then stir at 80 DEG C, until reacted (about 5 hours).Reaction mixture pours in 150mL frozen water, uses the CH of 50mL × 3 ₂cl ₂extraction, merges extraction phase, uses salt water washing, anhydrous sodium sulfate drying.Suction filtration removing siccative, filtrate is evaporate to dryness on a rotary evaporator, and the resistates obtained uses column chromatography purification, obtains compound IV-1, white solid, ESI-MS, m/z=223 ([M+H] ⁺).

B. the synthesis of compound V-1

Compound IV-1 (1.55g, 7mmol) is dissolved in 15mL dehydrated alcohol, and stirred at ambient temperature adds the hydrazine hydrate of 1mL 80%, and gained reaction mixture then at room temperature stirs, until reacted (TLC, within 5 hours).The direct evaporate to dryness on a rotary evaporator of reaction mixture, the direct column chromatography purification of the resistates obtained, obtains compound V-1, white solid, ESI-MS, m/z=209 ([M+H] ⁺).

C. the synthesis of Compound I-1

Compound V-1 (0.83g, 4mmol) with triethylamine (1.21g, 12mmol) be dissolved in the methylene dichloride of 10mL drying, ice-water bath cooling is lower stirs, the solution that the methylene dichloride slowly dripping VI-1 (0.88g, 4mmol) and 1mL drying is made, after dropwising, reaction mixture at room temperature stirs 1 hour, and TLC shows reaction to be completed.Reaction mixture pours in 100mL frozen water, uses the CH of 50mL × 3 ₂cl ₂extraction, merges extraction phase, uses salt water washing, anhydrous sodium sulfate drying.Suction filtration removing siccative, filtrate is evaporate to dryness on a rotary evaporator, and the resistates obtained uses column chromatography purification, obtains Compound I-1, white solid, ESI-MS, m/z=394 ([M+H] ⁺).

the synthesis of embodiment 2 reference compound D-1

For further illustrating the beneficial effect of the compounds of this invention, applicant describes applicant's research but not yet disclosed Compound D-1 and pharmacological datum.

A. the synthesis of compound IV-1

Compound II per-1 (0.68g, 10mmol) with compound III (1.95g, 10mmol) be dissolved in the DMF of 20mL drying, stirred at ambient temperature, add solid carbonic acid potassium (4.15g, 30mmol) with 0.5g KI, then stir at 80 DEG C, until reacted (about 5 hours).Reaction mixture pours in 150mL frozen water, uses the CH2Cl2 extraction of 50mL × 3, merges extraction phase, use salt water washing, anhydrous sodium sulfate drying.Suction filtration removing siccative, filtrate is evaporate to dryness on a rotary evaporator, and the resistates obtained uses column chromatography purification, obtains compound IV-1, white solid, ESI-MS, m/z=183 ([M+H]+).

B. the synthesis of compound V-1

Compound IV-1 (1.28g, 7mmol) is dissolved in 15mL dehydrated alcohol, and stirred at ambient temperature adds the hydrazine hydrate of 1mL 80%, and gained reaction mixture then at room temperature stirs, until reacted (TLC, within 5 hours).The direct evaporate to dryness on a rotary evaporator of reaction mixture, the direct column chromatography purification of the resistates obtained, obtains compound V-1, white solid, ESI-MS, m/z=169 ([M+H]+).

C. the synthesis of Compound D-1

Compound V-1 (0.67g, 4mmol) with triethylamine (1.21g, 12mmol) be dissolved in the methylene dichloride of 10mL drying, ice-water bath cooling is lower stirs, the solution that the methylene dichloride slowly dripping VI-2 (0.71g, 4mmol) and 1mL drying is made, after dropwising, reaction mixture at room temperature stirs 1 hour, and TLC shows reaction to be completed.Reaction mixture pours in 100mL frozen water, uses the CH2Cl2 extraction of 50mL × 3, merges extraction phase, use salt water washing, anhydrous sodium sulfate drying.Suction filtration removing siccative, filtrate is evaporate to dryness on a rotary evaporator, and the resistates obtained uses column chromatography purification, obtains Compound D-1, white solid, ESI-MS, m/z=309 ([M+H]+).

embodiment 3 Compound ira vitro is tested the excitement of GPR119

People-mouse chimera GPR119 the expression construct of front 198 amino acid of coding FLAG epitope tag, people GPR119 and amino acid whose 3 copies of the C-terminal 137 of mouse receptor being cloned into tsiklomitsin can in inducible vectors pcDNA5/FRT/TO (Invitrogen#V6520-20), and it comprises the marker of hygromycin resistance.By extremely being expressed by this construct stable integration in the genome of specific host cell system Flp-In-T-Rex-HEK293 (Invitrogen) of tetracycline repressor, realize the expression of receptor of precise hard_drawn tuhes.Once produce the clone of stable hygromycin resistance, cell is maintained at the 5%CO of humidification at 37 DEG C ₂in atmosphere and substratum.This substratum is made up of DMEM (Dulbecco ' s modified Eagle ' s medium) (DMEM, the Invitrogen) being supplemented with 2mM L-glutaminate, 10% foetal calf serum, 200 μ g/ml hygromycin B and 15 μ g/ml blasticidins (blasticidin).

Before carrying out cAMP accumulation mensuration 48 hours, stably express is fitted together to the cell of people/mouse GPR119 construct with 4 × 10 ³the density of cells/well is seeded in the solid blank (No. 35-6661st, BD) of the 384 poly-D-Lys coatings in hole, and makes it in 37 DEG C of 5%CO at humidification ₂atmosphere and be supplemented with 1 μ g/ml tsiklomitsin substratum in grow to bring out expression of receptor.On mensuration same day, removed substratum and in 37 DEG C at humidification 5%CO ₂atmosphere and 20 μ l/ holes measure damping fluid and (have Ca ²⁺and Mg ²⁺phosphate buffered saline (PBS), 12mM glucose, 0.1mM isobutyl-methyl-xanthine, 0.1% not fatty acids bovine serum albumin) in by cell incubation 50min, this mensuration damping fluid has expects that the compound added from the Concentrated stocks be dissolved in methyl-sulphoxide (DMSO) of concentration to obtain the ultimate density of 1%DMSO in mensuration.Use CisBio homogeneous phase time discrimination fluorescence (HTRF) to measure test kit, the code following manufacturers measures cAMP accumulation.In brief, in each hole, add 10 μ lcAMP-HTRF luciferase assay reagents separately, and by sample in incubated at room 40min.At 320nm fluorescence excitation and in 665nm and 620nm use Envision instrument (Perkin Elmer) measurement.Calculate the fluorescence ratio of 665/620 and the nanomolar concentration by being translated into the cAMP in each hole from cAMP typical curve interpolation.Excel/XLfit software (Microsoft and IDBS) is adopted to utilize four parameter logistic curve fit Equation for Calculating concentration-response curve and EC ₅₀.

Test result sees the following form.

Compound	EC 50(nM)
		Reference compound D-1	22.7
The compounds of this invention I-1	5.9

As can be seen from upper table result, compound of the present invention is good GPR119 agonist, can be used for preparation treatment type ii diabetes medicine.

Claims (3)

1. there is the compound of formula I structure,

2. synthesize the method for formula I described in claim 1:

Compound II per and compound III, in generation substitution reaction, generate compound IV; Compound IV and hydrazine hydrate react, and generate compound V; Compound V and compound VI are reacted, and generate I.

3. the application of formula I described in claim in preparation treatment diabetes B medicine.