CN108129465A - A kind of glyoxaline compound of 2- pyrazines derivatives substitution and its application in medicine for treating osteoporosis is prevented - Google Patents

Abstract

The invention discloses a kind of glyoxaline compound of 2 pyrazines derivatives substitution, structural formula is

Description

The glyoxaline compound and its preventing osteoporosis that a kind of 2- pyrazines derivatives replace Application in drug

Technical field

The invention belongs to drug design and synthesis field, be related to a kind of substitution of 2- pyrazines derivatives glyoxaline compound and Its application in medicine for treating osteoporosis is prevented.

Technical background

Osteoporosis is a health problem that is worldwide, increasingly attracting people's attention.The whole world about 2 at present Hundred million people suffer from osteoporosis, incidence leapt to common disease, the 7th of frequently-occurring disease.Osteoporosis is that the elderly is most common Disease.It is more than in 55 years old postclimacteric women at the age, the people for having half suffers from the different bone of degree in the U.S. and western countries Matter osteoporosis, and suffer from osteoporosis person in the elderly of over-65s and be up to more than 95%.With the reach of science and the mankind Progress, the average life span constantly extend, and elderly population increase sharply, and the incidence of osteoporosis rapidly increases also with appearance. It is expected that the elderly to the year two thousand fifty whole world over-65s will increase to 15.55 hundred million by current 3.23 hundred million, osteoporosis is led at that time Cause hip fracture that number occurs also will increase to 6,260,000, wherein Asia, Latin America, the Middle East and African state by current 1,660,000 The patient of family will account for more than 70%.Osteoporosis causes huge society and financial burden, constitute one it is serious global Problem has caused the very big concern of countries in the world.The arrival of aging society makes the incidence of osteoporosis increase increasingly, sternly People’s lives quality is threatened again.

The osteoporosis epidemiological survey in 3 city of China Beijing, Shanghai and Chengdu is the results show that 60-69 Sui age bracket bone The loose incidence women of matter and male are respectively 60% and 30% or so.Population base of China is big, and sufferers of osteoporosis face has reached 60000000~80,000,000, this brings serious burden to family and society.According to another the people of statistics national over-65s in 2000 Have 1.3 hundred million, account for the 10.7% of national population, it has also become typical veteran form country.And the patient with osteoporosis is at these Account for about 90% in crowd, and the patient of hip fracture will have 12%~40% to die of various complication in 1 year.In survivor The human action for having 50% is inconvenient.This not only causes serious body and mind to patient and wrecks, while also increases to family and society huge Big manpower and financial resources burden.Therefore, prevention osteoporosis is anti-aging, extends the service life, ensures the one of the quality of life of the people A very urgent research topic.

Osteoclast is a kind of terminal differentiation multinuclear macrophage originating from candidate stem cell, is uniquely joined in bone metabolism With the cell of bone information, it is in bone tissue this microenvironment by the portion perimeter blood of derived from bone marrow（Spleen）Monocyte and monokaryon The osteoclastic precursor of macrophage merges.Key link in bone remodeling is the bone information of osteoclast, normal stablizing It is very important in bone homeostasis process.Some drugs such as bisphosphonates, calcitonin surround in anti-bone information drug Osteoclast is unfolded, and prevents to reduce bone information by inhibiting the activity of osteoclast or treats osteoporosis.

Invention content

One of the objects of the present invention is to provide a kind of glyoxaline compounds of 2- pyrazines derivatives substitution, and structural formula is such as Under：

。

Another object of the present invention is to provide a kind of application of compound in osteoclast increment drug is inhibited.

Another object of the present invention is to provide a kind of compound to prepare prevention and/or treatment medicine for treating osteoporosis In application.

Another object of the present invention is to provide a kind of synthesis of the glyoxaline compound of the 2- pyrazines derivatives substitution Route is：

。

Further, specific synthesis step is：

1）3- phenyl-1-propanols（Compound 1）With sulfonylation generation methanesulfonic acid 3- benzene occurs under paratoluensulfonyl chloride low temperature Base propyl ester（Compound 2）；

2）Compound 2 heats in the presence of an inorganic base with 1- (3- hy droxy furan -2- bases) second -1- ketone again reacts generation 1- (3- (3- Phenyl-propoxy) furans -2- bases) second -1- ketone（Compound 3）；

3）Bromo-reaction generation 2- bromo- 1- (3- (the 3- phenyl third of carbonyl α-carbon occurs with pyrrolidones hydrobromate for compound 3 Oxygroup) furans -2- bases) second -1- ketone（Compound 4）；

4）Substitution reaction generation 1- (3- (3- phenyl-propoxies) furans -2- bases) -2- (pyridine -2- base ammonia of amine occurs for compound 4 Base) ethane -1- ketone（Compound 5）；

5）The acylation reaction generation 5- methyl-N- (2- oxos -2- of amine occur with 5- methylpyrazine -2- carbonyls chlorine for compound 5 (3- (3- phenyl-propoxies) furans -2- bases) ethyl)-N- (pyridine -2- bases) pyrazine -2- formamides（Compound 6）；

6）Cyclization generation 2- methyl -5- ((4- (3- (3- phenyl-propoxies) furans -2- bases) -1- (pyrroles occur again for compound 6 Pyridine -2- bases) -1H- imidazoles -2- bases) pyrazine（Compound 7）.

Further, synthesis step 1）Middle low temperature range is 0 ~ 10 DEG C, preferably 0 DEG C.

Further, synthesis step 2）In the inorganic base used can be sodium carbonate, potassium carbonate, cesium carbonate, saleratus, It is preferred that potassium carbonate.

Further, synthesis step 2）In heating temperature be 50 DEG C -100 DEG C, preferably 85 DEG C.

Obviously, the above according to the present invention according to the ordinary technical knowledge and means of this field, is not departing from this hair Under the premise of bright above-mentioned basic fundamental thought, the modification, replacement or change of other diversified forms can also be made.

Specific embodiment

Embodiment：

The synthesis of 1-1 methanesulfonic acid 3- phenylpropyl acrylates：

By 3- phenyl-1-propanols（Compound 1）(2.72g, 20mmol) is dissolved in dichloromethane (DCM) (25mL), adds in three second Amine (TEA) (4.05g, 40mmol) obtains mixture, and mixture then is cooled to 0 DEG C.Under stiring, delay into the mixture It is slow to add in paratoluensulfonyl chloride (MsCl) (5.72g, 30mmol), and reaction mixture is stirred 1 hour at 0 DEG C, Ran Hou It is stirred at room temperature 1 hour (being completed until detecting reaction by HPLC).After removing solvent, it is molten to add in saturated sodium bicarbonate water Liquid.Product is extracted with EtOAc (3 × 30mL) and is washed with sodium bicarbonate solution and water, then removes solvent in a vacuum After obtain methanesulfonic acid 3- phenylpropyl acrylates（Compound 2）, 3.86g, yield 90.0%.¹H-NMR (400 MHz, CDCl₃) δ: 1.82(m, 2H), 2.77(t, 2H), 3.16(s, 3H), 4.02(t, 2H), 7.15-7.26(m, 5H). ¹³C-NMR (125 MHz, CDCl₃) δ: 29.65, 34.34, 37.81, 67.65, 126.17, 128.33, 129.22, 142.31.LC-MS(ESI, pos, ion) m/z: 215[M+1]。

The synthesis of 1-2 1- (3- (3- phenyl-propoxies) furans -2- bases) second -1- ketone：

The compound 2 (3.86g, 18.01mmol) that above-mentioned synthesis step 1-1 is obtained is dissolved in anhydrous n,N-Dimethylformamide (DMF) in (25mL), 1- (3- hy droxy furan -2- bases) second -1- ketone (2.27g, 18.01mmol) and potassium carbonate are then added in (7.47g, 54.03mmol).Mixture at 85 DEG C is heated into 18 hours (being completed until detecting reaction by HPLC), then will It is cooled to room temperature.Saturated sodium bicarbonate aqueous solution is added in mixture, is extracted with ethyl acetate and uses sodium bicarbonate solution It is washed with water, after removing solvent in a vacuum, residue is obtained into off-white powder 1- (3- by purification by flash chromatography (3- phenyl-propoxies) furans -2- bases) second -1- ketone（Compound 3）3.91g, yield 88.9%.¹H-NMR (400 MHz, CDCl₃) δ: 2.05(m, 2H), 2.66(s, 3H), 2.77(t, 2H), 4.16(t, 2H), 6.28(d, 1H), 6.88(d, 1H), 7.15-7.26(m, 5H). ¹³C-NMR (125 MHz, CDCl₃) δ: 24.66, 29.95, 34.34, 70.26, 111.03, 126.17, 128.33, 129.22, 137.15, 142.31, 146.41, 152.18, 193.54. LC-MS(ESI, pos, ion) m/z: 245[M+1]。

The synthesis of the bromo- 1- of 1-3 2- (3- (3- phenyl-propoxies) furans -2- bases) second -1- ketone：

At 0 DEG C, the compound 3 (3.91g, 16.00mmol) that synthesis step 1-2 is obtained is dissolved in anhydrous MeOH (30mL), Then pyrrolidones hydrobromate (6.26g, 19.2mmol) is added in into this solution.Then by reaction mixture in nitrogen atmosphere Under stirred 4 hours at 0 DEG C, and be allowed to warm to room temperature and continue stirring until reaction is completed (to detect by TLC or HPLC aobvious Show).Then solvent is removed in a vacuum, the flash column chromatography isolated bromo- 1- of 4.48g yellow 2- (3- (the third oxygen of 3- phenyl Base) furans -2- bases) second -1- ketone（Compound 4）Powder, yield 86.7%.¹H-NMR (400 MHz, CDCl₃) δ: 2.05(m, 2H), 2.77(t, 2H), 4.16(t, 2H), 4.58(s, 2H), 6.25(d, 1H), 6.88(d, 1H), 7.15- 7.26(m, 5H). ¹³C-NMR (125 MHz, CDCl₃) δ: 29.95, 32.52, 34.34, 70.26, 111.03, 126.17, 128.33, 129.22, 134.87, 142.31, 146.41, 150.86, 191.83. LC-MS(ESI, pos, ion) m/z: 323[M+1]。

The synthesis of 1-4 1- (3- (3- phenyl-propoxies) furans -2- bases) -2- (pyridine -2- bases amino) ethane -1- ketone：

2-aminopyridine (0.94g, 10mmol) is dissolved in anhydrous DMF (20mL), then into solution add in DIEA (3.88g, 30mmol), stirring one adds in compound 4 (3.23g, 10mmol) after the meeting, by reaction mixture in a nitrogen atmosphere at room temperature Stirring is until reaction is completed（It is completed by TLC or HPLC detection reactions）.Then reaction mixture with cold water is diluted, uses acetic acid Ethyl ester extracts dilution, and the organic layer of merging is washed with brine and dried over sodium sulfate.It is obtained after solvent is evaporated in vacuo Required product 1- (3- (3- phenyl-propoxies) furans -2- bases) -2- (pyridine -2- bases amino) ethane -1- ketone（Compound 5）, then crude product by silica gel chromatography (2-4%MeOH/DCM) is purified, obtains purer compound 5,2.64g, production Rate 78.5%.¹H-NMR (400 MHz, CDCl₃) δ: 2.05(m, 2H), 2.77(t, 2H), 3.00(s, 1H),4.16 (t, 2H), 5.00(s, 2H), 6.22(d, 1H), 6.81(t, 1H),6.88(d, 1H), 7.13(d, 1H),7.15- 7.26(m, 5H). ¹³C-NMR (125 MHz, CDCl₃) δ: 29.95, 34.34, 46.62, 70.26, 108.84, 110.44, 111.03, 126.17, 128.33, 129.22, 136.64, 138.85, 142.31, 146.41, 149.77, 156.49, 158.35, 182.94.LC-MS(ESI, pos, ion) m/z: 337[M+1]。

1-5 5- methyl-N- (2- oxos -2- (3- (3- phenyl-propoxies) furans -2- bases) ethyl)-N- (pyridine -2- bases) The synthesis of pyrazine -2- formamides：

Compound 5 (1.68g, 5mmol) is dissolved in anhydrous DCM (20mL) at 0 DEG C, TEA is then added in into solution (1.52g, 15mmol) is slow added into 5- methylpyrazine -2- carbonyls chlorine (2.35g, 15mmol).By reaction mixture in nitrogen It is stirred 4 hours at 0 DEG C under atmosphere, is then allowed to warm to room temperature, continue stirring until reaction is completed (by TLC or HPLC Detection reaction is completed).Then it removes solvent under reduced pressure, obtains crude product 5- methyl-N- (2- oxos -2- (3- (3- phenyl-propoxies) furans Mutter -2- bases) ethyl)-N- (pyridine -2- bases) pyrazine -2- formamides（Compound 6）, with acetone and ethyl alcohol recrystallization, obtain 2.03g refine after compound 6, yield 89.1%.¹H-NMR (400 MHz, CDCl₃) δ: 2.05(m, 2H), 2.59(s, 3H), 2.77(t, 2H), 4.15(t, 2H), 6.26(d, 1H), 6.43(s, 2H), 6.87(d,1H), 7.14- 7.25(m, 5H), 7.48(t, 1H), 7.85(t, 1H), 8.48(d, 1H), 8.59(s, 1H), 8.70(d, 1H), 9.77(s, 1H). ¹³C-NMR (125 MHz, CDCl₃) δ: 21.51, 29.95, 34.34, 46.89, 70.26, 111.03, 117.32, 118.09, 126.17, 128.33, 129.22, 137.21, 138.68, 139.48, 142.31, 143.54, 146.41, 150.27, 150.34, 150.62, 153.89, 157.15, 158.20, 179.97. LC-MS(ESI, pos, ion) m/z: 457[M+1]。

1-6 2- methyl -5- ((4- (3- (3- phenyl-propoxies) furans -2- bases) -1- (pyridine -2- bases) -1H- imidazoles -2- Base) pyrazine synthesis：

Compound 6 (2.03g, 4.45mmol) obtained above is dissolved in acetic acid (20mL), then under stiring to above molten Ammonium acetate (6.86g, 89.00mmol) is added in liquid, reaction mixture is stirred overnight at 90 DEG C.Then by reaction mixture It is cooled to room temperature, and is neutralized with saturated sodium bicarbonate solution.Then after being extracted with EtOAc, then it is passed through into silica gel column chromatography (4- 6%MeOH/DCM is eluted) purifying, obtain product 2- methyl -5- ((4- (3- (3- phenyl-propoxies) furans -2- bases) -1- (pyrroles Pyridine -2- bases) -1H- imidazoles -2- bases) pyrazine（Compound 7）Off-white powder 1.75, yield 90%.¹H-NMR (400 MHz, CDCl₃) δ: 2.05(m, 2H), 2.59(s, 3H), 2.77(t, 2H), 4.16(t, 2H), 6.02(d, 1H), 7.06(d, 1H), 7.14-7.25(m, 5H), 7.39(t, 1H), 7.89(d, 1H), 8.00(t, 1H), 8.38(s, 1H), 8.48(s, 1H), 8.50(d, 1H), 8.86(s, 1H). ¹³C-NMR (125 MHz, CDCl₃) δ:21.51, 29.95, 34.34, 36.36, 70.26, 110.25, 123.60, 125.34, 126.17, 127.39, 128.33, 129.22, 133.51, 139.48, 142.12, 142.30,142.31, 142.87, 142.92, 148.40, 149.00, 149.03, 151.97, 152.25, 152.43. LC-MS(ESI, pos, ion) m/z: 438[M+1]。

Test example 1：To the increment inhibiting effect of osteoclast

First, experimental method

1st, osteoclast is separately cultured

Neck is drawn to put to death the Wistar rats in 24 h of birth, after distilled water cleans 1 min, 75% ethyl alcohol impregnates 2min, detaches stock Bone, shin bone and humerus carefully remove the soft tissue of bone surface, wipe out epiphysis, diaphyseal portion is washed 2 times with α-MEM culture solutions, is put into In cold α-MEM culture solutions, backbone stringer is cut off with microsurgical scissors, marrow inner cavity surface sclerotin is gently scraped with scalpel Enter in culture solution, then sclerotin fragment 2min is gently blown and beaten with round end suction pipe, the osteoclast being attached on bone cips is poured into training In nutrient solution, 15s is stood, draws upper cell suspension, it is about 10 that cell density, which is made,⁶The cell suspension of a/ml is inoculated in preset In 24 porocyte culture plates of coverslip, CO is put₂Incubator（5% CO₂, 37 DEG C）After cultivating 2h, the culture solution in culture hole is sucked out Not adherent heteroproteose cell is removed, is gently rinsed 2 times with the α-MEM of pre-temperature, be grouped and continues to cultivate after being administered according to grouping, Liquid 50% is changed per 48h.Above-mentioned α-MEM culture solutions are purchased from Hyclone companies.

2nd, grouping and administration

Experiment is divided into medicine group to be measured and control group, is used in medicine group to be measured and contains 1 μm of ol/L untested compound（Corresponding embodiment Middle difference compound）α-MEM culture solutions, with the α-MEM culture solutions for not containing 1 μm of ol/L untested compound in control group.

3rd, osteoclast morphologic observation

Carry out somatoscopy after 48h to cell under inverted microscope, while each glass creep plate takes 5 different 200 at random Times visual field counts apocyte（3 or 3 or more）Sum it is osteoclastic as the time point on each slide（Sample）Cell is total Number.

4th, statistical disposition

As a result it is represented with mean ± S.D., all data are handled using SPSS16.0 softwares, multiple-group analysis one way ANOVA variance analyses, comparison among groups are examined with q；Two groups are compared and are examined with t.P <0.05 is statistically significant for difference.

5th, experimental result

1 osteoclast of table counts（A/piece,）

Grouping	Number
		Control group	28.1±3.6
Compound 6	15.6±3.8*
		Compound 7	11.2±3.3*

Note：Compared with the control group, * P ＜ 0.05

Compared with the control group, amount of osteoclast has prepared each compound in embodiment it can be seen from upper table result Different degrees of reduction, and somatoscopy is carried out to cell under inverted microscope and finds the visible cell space of control group osteoclast It is very big, it is clear-cut, in pan-fried oily egg shape, dumbbell shape, funnel type, chopper type etc., there are a large amount of pseudopodium to stretch around, nucleus is very It is more, generally 8~15.And there is cell space diminution in the osteoclast of compound 6 and compound 7 in experimental group, pseudopodium disappears It loses, the apoptosis phenomenon such as karyopycnosis.Illustrate that each compound of experimental group is inhibited to the increment of osteoclast.

Embodiment 2：To the preventive and therapeutic effect of vitamin A acid modeling osteoporosis rat

First, drug is prepared

1st, vitamin A acid solution accurately weighs vitamin A acid 3.0g, and 0.5% CMC-Na solution is added to obtain 15mg/ml solution to 200ml, standby With；

2nd, Ipriflavone solution takes Ipriflavone 6.0g, adds DMSO 12ml, adds in 0.5% CMC-Na solution to 300ml, obtains 20mg/ml solution, it is spare；

3rd, testing compound solution is prepared by the compound in corresponding embodiment, accurately weighs 0.3g, adds DMSO 4ml, then add Enter 0.5% CMC-Na solution to 100ml, obtain 3mg/ml solution, it is spare.

2nd, experimental method

Healthy female SD rats are randomly divided into 4 groups, respectively control group by weight（N=5）, model group（N=10）, it is positive right According to group（Ipriflavone 100mg/kg, n=10）, several medicine groups to be measured（15mg/kg, n=10）.After experiment starts, per in the sky Noon other each group gavages in addition to control group give vitamin A acid 75mg/kg（5ml/kg）, control group gives isometric 0.5% CMC- Na, afternoon positive controls and medicine group to be measured give relative medicine solution respectively, control group and model group are given in equal volume 0.5% CMC-Na, continuous 2 weeks.

The left femur of animal is taken to use toy bone strength analyzer（YLS-16A toy bone strength analyzers, Huaibei Zhenghua Biological Instrument Co., Ltd.）Measure the bone breaking force of femur, each sample be placed on position on instrument and Direction is consistent；Residual bone is collected after measure, is wrapped up with the wet gauze that physiological saline impregnates, is sealed after -20 DEG C of preservations For measuring bone calcium and bone phosphorus.

The grey method assay of bone calcium, bone phosphorus：Room temperature naturally to thaw, 120 DEG C of baking 12h, claim backbone weight in baking oven（W1）, Bone is placed in 600 DEG C of muffle furnace ashing 3h again, makes the white bulk of bone, and claim bone ash weight（W2）.The bone ash that will be fully ground again （0.1g）It is dissolved in excess（4ml）3% HNO₃In, fully extract（It places 2 days and shakes up）, grey method measure calcium, phosphorus content.

3rd, experimental result

As a result it is represented with mean ± S.D., all data are handled using SPSS16.0 softwares, multiple-group analysis one way ANOVA variance analyses, comparison among groups are examined with q；Two groups are compared and are examined with t.P <0.05 is statistically significant for difference.

2 the compounds of this invention of table is to bone breaking force and bone calcium, the influence of bone phosphorus

Note：Compared with model group, * P ＜ 0.05

By upper table result it is found that rat to vitamin A acid modeling, the compounds of this invention 6 and compound 7 can increase bone breaking force Size, and increase the content of bone calcium and bone phosphorus, illustrate that the compounds of this invention can have osteoporosis prevention and/or control Treatment acts on, and osteoporosis new construction drug can be used as to carry out more in-depth study and exploitation.

Claims (4)

1. a kind of glyoxaline compound of 2- pyrazines derivatives substitution, structural formula are as follows：

。

2. application of the compound as described in claim 1 in osteoclast increment drug is inhibited.

3. application of the compound as described in claim 1 in prevention and/or treatment medicine for treating osteoporosis is prepared.

4. the synthetic route of compound as described in claim 1 is：

。