CN107118212B - 1- pyridine -6- methoxyl group -9- (3- iodine benzyl)-B-carboline, synthesis and application - Google Patents
1- pyridine -6- methoxyl group -9- (3- iodine benzyl)-B-carboline, synthesis and application Download PDFInfo
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- CN107118212B CN107118212B CN201710385798.XA CN201710385798A CN107118212B CN 107118212 B CN107118212 B CN 107118212B CN 201710385798 A CN201710385798 A CN 201710385798A CN 107118212 B CN107118212 B CN 107118212B
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
Abstract
The present invention relates to a kind of 'Beta '-carboline compound and its synthetic method and applications, belong to pharmaceutical technology field.The compound structure is shown in formula I, shows that the anti-kidney fibrosis activity of the Compound ira vitro is significant through preliminary experiment, has preferable potential medical value, can be used for the preparation of various anti-kidney fibrosis, anti-chronic kidney disease drug.
Description
[technical field]
The present invention relates to pharmaceutical technology fields, and in particular to a kind of 'Beta '-carboline compound and its synthetic method and application.
[background technique]
Kidney fibrosis caused by chronic kidney disease is lost with renal function progressive, extracellular matrix (ECM) is largely built up
It is characterized, mainly includes glomerulosclerosis and Tubulointerstitial fibrosis.Kidney region fibrosis is almost all chronic renal diseases
The final result of development is in progress closely related with renal failure, is the Etiological for leading to end stage renal failure.If early stage is right
Renal fibrosis carries out therapeutic intervention, it will help delaying chronic kidney trouble progress greatly reduces the incidence of end-stage renal disease
And its complication, mitigate the social economical burden caused by it.
The study found that transforming growth factor-β (TGF-β) and its Smad signal path mediated are in kidney region fibrosis
It plays an important role, it is considered to be most critical, strongest fibrosis inducible factor participate in the links of kidney fibrosis.It
Overexpression being capable of mesangial cells stimulated, a large amount of collagens (Collagen) of Stromal fibroblasts synthesis, fibronectin
(FN) it simultaneously activates the secretion for promoting ECM at myofibroblast with laminin (LN), stimulation fibroblast proliferation and sinks
Product.TGF-β can also mediate transdifferentiation (EMT) of the renal cells to interstitial cell.It clinically there is no at present effective anti-
The treatment method of renal fibrosis.Antagonism TGF-β and its signal path of mediation inhibit ECM synthesis or promote its degradation, may
An effective approach is provided to find treatment renal fibrosis.
Beta-carboline alkaloid is the one kind separated from Xinjiang medicinal plant harmel (Peganumharmala)
Active constituent.Pharmacological activity with wide spectrum, such as antianxiety, antidepression, anti-spasm and antitumor, anti-malarial, anti-parasitism
Worm, anti-AIDS etc..It is long-standing come treating cancer with seed of peganum harmala in Chinese herbal medicine formula.Beta-carboline alkaloid
Structural modification is always by the concern of researcher.It is generally believed that the planar conjugate structure of most of beta-carboline alkaloids
And different substituent group (such as benzyl (- C6H5), methoxyl group (- OCH3)) presence and its anti-tumor activity there is significant close
System.Mainly include at present the following aspects to the research of beta-carboline alkaloid: first is that based on natural product chemistry research from
It is found in the plant of not equal category and carries out separating-purifying；Second is that being carried out by methodology of organic synthesis to beta-carboline alkaloid complete
Synthesis is semi-synthetic, modifies its structure.It in addition is then that molecule mechanism is carried out to its pharmacological activity (such as anti-tumor activity)
Research.It is such as inserted into DNA, inhibit topoisomerase, inhibit CDK etc..But from the point of view of current progress, beta-carboline alkaloid
Content in natural plants is generally all relatively low, and extraction is relatively complicated, is unfavorable for furtheing investigate it, and
Though organic semisynthesis has certain advantage on yield, limited by cost is produced, therefore at present to B-carboline
The expansion research of Alkaloid is still inadequate.Such compound has good development prospect, but yet there are no 1- pyrrole
Pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline and its relevant report of synthesis and application.
[summary of the invention]
Goal of the invention of the invention is: in view of the above problems, providing a kind of 'Beta '-carboline compound and its synthesis
Methods and applications.Production process environmental protection of the present invention, production cost is low, and operational safety is high, and reaction condition is mild, easy to operate；
Obtained 'Beta '-carboline compound can be used for preparing anti-kidney fibrosis drug and/or anti-chronic kidney disease drug.
To achieve the goals above, The technical solution adopted by the invention is as follows:
A kind of compound or its pharmaceutically acceptable salt of Formulas I:
The chemical name of compound shown in above-mentioned Formulas I are as follows: 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline.Point
Son amount are as follows: 491.05.
The present invention also provides the synthetic method of the compound of the Formulas I, include the following steps: for sodium hydride to be dissolved in N, N-
In dimethylformamide, compound 1- pyridine -6- methoxy-p-carboline and 3- iodobenzene first bromine shown in formula II, Yu Chang is then added
Temperature or heating condition under one pot reaction to get the Formulas I compound.
The synthetic route of above-mentioned synthetic method is as follows:
In the step of compound synthesis method shown in above-mentioned Formulas I, raw material 1- pyridine -6- methoxy-p-carboline and another original
Expect that the ratio between amount of substance of 3- iodobenzene first bromine is usually 1:1.5 or 1:1, the preferably mole of 3- iodobenzene first bromine is slightly larger than 1- pyrrole
The amount of the substance of pyridine -6- methoxy-p-carboline is carried out with reacting fully.
Further, the reaction carries out under 40 DEG C to organic solvent of reflux temperature.Whether reaction can adopt completely
With thin-layer chromatography (TLC) tracing detection, usually controlling the reaction time is that 20min~2h is appropriate.The mode of reaction preferably uses
Back flow reaction, when using back flow reaction, reaction is more preferably carried out under the reflux temperature of organic solvent.
Synthetic method of the present invention, further includes separation and purification step, and the separation and purifying are by one pot reaction
Products therefrom is added dropwise alkaline solution and is adjusted to neutral or alkalinity, is extracted with ethyl acetate or chloroform, collects organic phase, removes solvent,
Then it is purified by the way of silica gel column chromatography or recrystallization；When being purified by the way of silica gel column chromatography,
The eluant, eluent that petroleum ether and ethyl acetate with volume ratio for 1000:50~100 form is eluted, and eluent removes solvent,
Obtain object after purification.
The present invention also provides the compound or its pharmaceutically acceptable salt prepare anti-kidney fibrosis drug and/
Or the application in anti-chronic kidney disease drug.
The present invention also provides a kind of pharmaceutical compositions comprising the compound described in claim 1 of therapeutically effective amount
Or its pharmaceutically acceptable salt and pharmaceutically acceptable auxiliary material.
In conclusion by adopting the above-described technical solution, the beneficial effects of the present invention are:
Compared with prior art, the present invention provides a kind of 'Beta '-carboline compound and its synthetic method and applications.Inventor
The anti-kidney fibrosis of the Compound ira vitro is shown to anti-kidney fibrosis, the anti-chronic kidney disease activity research of compound of the present invention
It is active significant, there is preferable potential medical value, can be used for the preparation of various anti-kidney fibrosis, anti-chronic kidney disease drug.
[Detailed description of the invention]
Fig. 1 is the hydrogen nuclear magnetic resonance spectrogram of final product made from the embodiment of the present invention 1；
Fig. 2 is the carbon-13 nmr spectra figure of final product made from the embodiment of the present invention 1；
Fig. 3 is the electrospray ionization mass spectrum spectrogram of final product made from the embodiment of the present invention 1；
Fig. 4 is the cell protein immunoblot experiment figure of final product made from the embodiment of the present invention 1.
[specific embodiment]
The preparation of 1 structure of embodiment compound shown in formula I
1) by the NaH of 200mg (8.3mmol), be dissolved in 1mLN, in dinethylformamide, stir 10min, and gradually plus
1- pyridine -6- the methoxy-p-carboline for entering 275mg (1mmol), is stirred at room temperature 10min, 445mg is then added dropwise again
The 3- iodobenzene first bromine of (1.5mmol), room temperature the reaction was continued about 20min；Products therefrom is adjusted with the ammonium hydroxide of 300ml after reaction
It to neutrality, then is extracted with ethyl acetate, collects organic phase, solvent is removed under reduced pressure, obtains the crude product of yellow powder；
2) crude product is using petroleum ether: the solvent that ethyl acetate=1000:100 (volume ratio) forms is as eluant, eluent, silica gel
Column chromatography, obtains faint yellow fluffy solid (400mg, yield 81.6%, purity 99.92%).
Nuclear-magnetism characterization is carried out to the faint yellow fluffy solid of gained, nuclear magnetic resonance spectroscopy and carbon spectrum are respectively such as Fig. 1 and 2 institute
Show.
1HNMR (400MHz, CDCl3) δ 8.55 (d, J=5.2Hz, 1H), 8.53 (s, 1H), 8.05 (d, J=5.2Hz,
1H), 7.74 (td, J=7.6,1.8Hz, 1H), 7.68 (d, J=7.8Hz, 1H), 7.65 (d, J=2.3Hz, 1H), 7.32
(ddd, J=7.4,4.9,1.3Hz, 1H), 7.28 (d, J=8.9Hz, 1H), 7.23 (dd, J=9.0,2.4Hz, 1H), 6.60
(tdd, J=9.2,7.0,2.1Hz, 1H), 6.08 (td, J=8.0,2.4Hz, 1H), 5.52 (s, 2H), 3.96 (s, 3H)
13CNMR (101MHz, CDCl3) δ 157.61,154.69,148.16,142.76,138.15,137.53,
136.69,134.71,131.41,124.75,123.13,122.20,121.79,120.97,118.93,114.65,
111.96,111.78,110.90,103.58,56.04,29.71.
Mass spectrum and efficient liquid phase chromatographic analysis are carried out to gained pale yellow powder product, it is as a result as shown in Figure 3 respectively.
Accordingly, it can be determined that above-mentioned orange powder product are as follows: 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline,
Chemical structural formula is as shown in following formula I:
The preparation of 2 structure of embodiment compound shown in formula I
1) by the NaH of 400mg (16.6mmol), be dissolved in 1mLN, in dinethylformamide, stir 10min, and gradually plus
1- pyridine -6- the methoxy-p-carboline for entering 550mg (2mmol), is stirred at room temperature 10min, 445mg is then added dropwise again
The 3- iodobenzene first bromine of (1.5mmol), room temperature the reaction was continued about 20min；Products therefrom is adjusted to 400ml ammonium hydroxide after reaction
Neutrality is extracted with ethyl acetate, and collects organic phase, solvent is removed under reduced pressure, obtains the crude product of yellow oily liquid；
2) crude product is using petroleum ether: the solvent that ethyl acetate=1000:80 (volume ratio) forms is as eluant, eluent, silicagel column
Chromatography, obtains faint yellow fluffy solid (381mg, yield 51.6%, purity 99.90%).
The faint yellow fluffy solid of gained is characterized by nuclear-magnetism, is determined as 1- pyridine -6- methoxyl group -9- of the present invention
(3- iodobenzyl) B-carboline.
The preparation of 3 structure of embodiment compound shown in formula I
1) by the NaH of 200mg (8.33mmol), be dissolved in 5mLN, in dinethylformamide, stir 10min, and gradually plus
1- pyridine -6- the methoxy-p-carboline for entering 275mg (1mmol), is stirred at room temperature 10min, 593mg is then added dropwise again
The 3- iodobenzene first bromine of (2mmol), room temperature the reaction was continued about 20min；Gained reactant is adjusted to 400ml ammonium hydroxide after reaction
Neutrality is extracted with ethyl acetate, and collects organic phase, solvent is removed under reduced pressure, obtains the crude product of yellow oily liquid；
2) crude product is using petroleum ether: the solvent that ethyl acetate=1000:50 (volume ratio) forms is as eluant, eluent, silicagel column
Chromatography, obtains faint yellow fluffy solid (440mg, yield 89.8%, purity 99.95%).
The faint yellow fluffy solid of gained is characterized by nuclear-magnetism, is determined as 1- pyridine -6- methoxyl group -9- of the present invention
(3- iodobenzyl) B-carboline.
The preparation of 4 structure of embodiment compound shown in formula I
1) by the NaH of 200mg (8.33mmol), be dissolved in 8mLN, in dinethylformamide, stir 10min, and gradually plus
1- pyridine -6- the methoxy-p-carboline for entering 275mg (1mmol), is stirred at room temperature 10min, 297mg is then added dropwise again
The 3- iodobenzene first bromine of (1mmol), room temperature the reaction was continued about 20min；Gained reactant is adjusted to 400ml ammonium hydroxide after reaction
Neutrality is extracted with ethyl acetate, and collects organic phase, solvent is removed under reduced pressure, obtains the crude product of yellow oily liquid；
2) crude product is using petroleum ether: the solvent that ethyl acetate=1000:50 (volume ratio) forms is as eluant, eluent, silicagel column
Chromatography, obtains faint yellow fluffy solid (375mg, yield 76.5%, purity 99.94%).
The faint yellow fluffy solid of gained is characterized by nuclear-magnetism, is determined as 1- pyridine -6- methoxyl group -9- of the present invention
(3- iodobenzyl) B-carboline.
The preparation of 5 structure of embodiment compound shown in formula I
1) it by the NaH of 200mg (8.33mmol), is dissolved in 10mLN, in dinethylformamide, stirs 10min, and gradually
1- pyridine -6- the methoxy-p-carboline of 275mg (1mmol) is added, 20min is stirred at room temperature, 593.mg is then added dropwise again
The 3- iodobenzene first bromine of (2mmol), room temperature the reaction was continued about 40min；Gained reactant is adjusted to 500ml ammonium hydroxide after reaction
Neutrality is extracted with ethyl acetate, and collects organic phase, solvent is removed under reduced pressure, obtains the crude product of yellow oily liquid；
2) crude product is using petroleum ether: the solvent that ethyl acetate=1000:100 (volume ratio) forms is as eluant, eluent, silica gel
Column chromatography, obtains faint yellow fluffy solid (327mg, yield 66.7%, purity 99.92%).
The faint yellow fluffy solid of gained is characterized by nuclear-magnetism, is determined as 1- pyridine -6- methoxyl group -9- of the present invention
(3- iodobenzyl) B-carboline.
The preparation of 6 structure of embodiment compound shown in formula I
1) by the NaH of 200mg (8.3mmol), be dissolved in 1mLN, in dinethylformamide, stir 10min, and gradually plus
1- pyridine -6- the methoxy-p-carboline for entering 275mg (1mmol), is stirred at room temperature 10min, 445mg is then added dropwise again
The 3- iodobenzene first bromine of (1.5mmol) is heated to 40 DEG C the reaction was continued about 20min；Products therefrom is with 350ml's after reaction
Ammonium hydroxide is adjusted to alkalinity, then is extracted with chloroform, collects organic phase, solvent is removed under reduced pressure, obtains the crude product of yellow powder；
2) solvent for anhydrous methanol and the methylene chloride composition that crude product is 1000:100 with volume ratio is recrystallized, and is obtained
To faint yellow fluffy solid (412mg, yield 82.5%, purity 99.96%).
The faint yellow fluffy solid of gained is characterized by nuclear-magnetism, is determined as 1- pyridine -6- methoxyl group -9- of the present invention
(3- iodobenzyl) B-carboline 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline.
The anti-kidney of 7 structure of embodiment compound 1- pyridine -6- methoxyl group -9- shown in formula I (3- iodobenzyl) B-carboline
Fibrosis experiment
One, 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline kidney fibroblast NRK-49F normal to rat
Proliferation inhibition activity experiment:
1, cell strain and cell culture
The normal kidney fibroblast NRK-49F cell strain of rat is selected in this experiment.
Cell strain culture used calf serum containing 10wt%, 100U/mL penicillin, 100U/mL streptomysin DMEM/F-
In 12 culture solutions, sets in 37 DEG C of incubators of 5%CO2 containing volumetric concentration and cultivate.
2, the preparation of untested compound
1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline used is that product is made in the embodiment of the present invention 1, by
Obtained by column Chromatographic purification, purity >=95%.After test-compound to be measured is dissolved with DMSO, then with d=0.22 μm of diameter
Filtering with microporous membrane degerming is placed at 4 DEG C and saves.Its DMSO liquid storage (concentration 0.002mol/L) is passed through into serum-free DMEM/
F-12 culture medium is successively diluted to five concentration gradients, respectively 20,10,5,2.5,1.25 μm of ol/L, wherein cosolvent DMSO
Final concentration≤1%.The target product under various concentration is tested first for the inhibiting rate of NRK-49F cell Proliferation, is considered as primary dcreening operation
As a result；Target product under different gradient concentrations is tested respectively again to the Proliferation Ability degree of NRK-49F cell, to the Fitting Calculation
Half-inhibitory concentration, i.e. IC50 value.
3, cell growth inhibition test (mtt assay)
(1) cell of logarithmic growth phase is configured to after trypsin digestion with the culture solution containing 10% calf serum
Concentration is the cell suspension of 5000/mL, is inoculated in 96 well culture plates with every 180 μ L of hole, makes cell density to be measured to 1000
~10000/hole (edge hole is filled with sterile PBS)；
(2) 5%CO2,37 DEG C of incubation 12h change serum-free DMEM/F-12 culture solution and continue to cultivate after cell is adherent
The 20 μ L of drug of a certain concentration gradient is added in 12h, every hole, and each concentration gradient sets 5 multiple holes；
(3) 5%CO2,37 DEG C are incubated for 48 hours, observe under inverted microscope；
(4) the MTT solution (5mg/mLPBS, i.e. 0.5%MTT) of 10 μ L is added in every hole, continues 4~8h of culture；
(5) culture is terminated, culture solution in hole is carefully sucked, every hole is added 200 μ LDMSO and sufficiently dissolves first a ceremonial jade-ladle, used in libation precipitating, oscillation
It with wavelength is 570nm in microplate reader, reference wavelength is the OD value that 450nm measures each hole after device mixes；
(6) it is arranged zeroing hole (culture medium, MTT, DMSO) simultaneously, (the drug dissolution of cell, same concentrations is situated between control wells
Matter, culture solution, MTT, DMSO).
(7) according to the OD value (OD value) measured, to judge living cells quantity, OD value is bigger, and cell activity is stronger.
Utilize formula:
Calculate the inhibiting rate of compounds on cell growth.Its test result is as shown in the following Table 1.
Table 1: compound is under various concentration to the growth inhibition ratio (%) of NRK-49F cell strain
It is further fitted by inhibiting rate data of the SPSS software to three concentration gradients, finds out product to NRK-
The half-inhibitory concentration (IC50 value, unit μm ol/L) of 49F cell strain, compound for NRK-49F cell strain IC50 value such as
Shown in table 2.
Table 2: IC50 value (μM) of the compound to NRK-49F cell strain
From the point of view of testing in vitro result, 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline to the normal kidney of rat at
Fibrocyte NRK-49F cell strain all shows significant proliferation inhibition activity, is measured with mtt assay 20,10,5,2.5,1.25
Inhibiting rate is respectively 69.27%, 54.46%, 22.90%, 14.29%, 13.06% when μm ol/L concentration, and IC50 is calculated
Value is 10.5 μM.
Two, 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline is to Mouse Kidney at brotic cells NRK-49F's
The protein immunoblotting of kidney fibrosis GAP-associated protein GAP is tested:
The present invention stimulates NRK-49F cell with TGF-β 1 (2ng/ml), establishes external fibrosis model, while difference is added
Compound shown in above-mentioned (I) of concentration (5 μM, 10 μM, 15 μM), after collective effect 48 hours, by extract proteins after cell cracking,
Carry out protein immunoblotting experiment, analysis fibronectin (Fibronectin, FN), α-smooth muscle actin (α-SMA),
The expression of the albumen such as collagen I (Collagen I), concrete operations are as follows:
1. protein extraction
(1) inoculating cell: the good NRK-49F cell of selection growth conditions is planted in the culture dish that diameter is 10cm,
37 DEG C, the constant incubator culture 12h that CO2 concentration is 5% are placed in, serum-free DMEM/F-12 culture solution is changed and continues to cultivate 12h.
(2) it is administered: when the adherent proliferation of cell is up to logarithmic growth phase, being separately added into the TGF-β 1 of 2ng/ml and not androgynous
Product, the chemical compounds I (being made by embodiment 1) that concentration is 2 × 10-3M, are incubated in constant incubator after being sufficiently mixed uniformly
48h。
(3) it collects cell: after cell administration handles 48h, being 0.25% trypsin digestion attached cell with concentration, turn
It moves in 15mL centrifuge tube, goes to centrifuge, 2000rpm is centrifuged 10min.
(4) cell cracking: distinguished with cell pyrolysis liquid (RIPA): PMSF (phenylmethylsulfonyl fluoride)=100:1 volume ratio
It is added, mixes suspension cell, be placed in 30~50min of cracking under low temperature environment on ice.
(5) collect albumen: by cracked 4 DEG C of cell suspension low temperature, 12000r is centrifuged 10min；Supernatant BCA albumen
(enhanced) the progress protein quantification of concentration measuring kit, -80 DEG C of storages.
2.SDS-PAGE electrophoresis
(1) preparation of glue and separation gel is concentrated: according to experiment needs, preparing the concentration of 10%, 12% separation gel and 5%
Glue.Separation gel is poured into vertical glass slot, carries out fluid-tight with distilled water.After separation gel solidification, distilled water is discarded, then will concentration
Glue pours into vertical glass slot insertion comb, solid wait be gelled.After gelling to be separated is solid, comb is extracted, vertical electrophoresis device is put into
Subsequent plus sample to be tested is waited to carry out electrophoresis in electrophoresis tank.
(2) loading and electrophoresis: take 20 μ g samples is 4:1 dilution proportion with 5 × sample-loading buffer by volume, in boiling water bath
5min is heated, 5 μ L albumen pre-dyed are added on the left of offset plate in loading after cooling.Electrophoresis liquid is added, electrophoresis tank is filled, by concentration glue
60V, 60min, separation gel 80V, 160min carry out electrophoresis.
(3) transferring film: glass plate is taken out, gel is removed and is dipped in transferring film buffer balances 10min；According to colored pre-dyed
Protein band carries out cutting glue.Take 8 filter paper be cut into transferring film sponge same size, soaked in transferring film buffer spare；It takes
Suitable pore size (0.45 μm or 0.22 μm) cellulose acetate film (PVDF), is cut into the PAGE glue same size with cutting.
Take 8 filter paper be cut into transferring film sponge same size, soaked in transferring film buffer spare；Take suitable pore size (0.45 μ
M or 0.22 μm) cellulose acetate film (PVDF), it is cut into the PAGE glue same size with cutting.Transferring film is folded up into transferring film slot
In, transferring film buffer is added, constant current 200mA carries out transferring film electrophoresis.To the end of transferring film, glue can be contaminated with Coomassie brilliant blue, with the beautiful spring
Red dye liquor contaminates film, detects transferring film effect；After pvdf membrane is washed with clear water, next step operation is carried out.
(4) be immunoreacted: to the end of transferring film, pvdf membrane closes 30min with quick closure fluid-tight again after being rinsed with PBS.PBST
5min is rinsed, the primary antibody fibronectin (Fibronectin, FN) by proper proportion configuration, α-smooth muscle actin are gone to
(α-SMA), collagen I (Collagen I) and collagen III (COL3A1)) in solution, 4 DEG C of overnight incubations.It is dilute with secondary antibody
Liquid dilution secondary antibody is released, about 1h is incubated on decolorization swinging table at room temperature, carries out chemiluminescence reaction after being rinsed with PBST and PBS.
(5) chemical illuminating reagent (the super quick luminescent solution of superECL, the limited public affairs of Puli's lema gene technology chemiluminescence: are taken
Department), it comes into full contact with memebrane protein face, develops in chemiluminescence imaging system.
Experimental result is as shown in Figure 4.
From the point of view of anti-kidney fibrosis test result, 1- pyridine -6- methoxyl group -9- (3- iodobenzyl) B-carboline can significantly press down
The expression of FN, α-SMA, COL3A1 of the NRK-49F cell that TGF-β 1 processed induces, and be in dose-dependence, show the chemical combination
Anti- kidney fibrosis activity is significant outside object, has preferable potential medical value, can be used for various anti-kidney fibrosis, anti-chronic renal
The preparation of popular name for drug.
10 pharmaceutical composition of embodiment
| Compound shown in Formulas I provided by the invention | 15% |
| Starch | 40% |
| Microcrystalline cellulose | 25% |
| Carboxymethyl starch is received | 19% |
| Magnesium stearate | 1% |
The above drug and auxiliary material grinding and sieving, mix to obtain the final product.
11 pharmaceutical composition of embodiment
| Compound shown in Formulas I provided by the invention | 8% |
| Starch | 47% |
| Microcrystalline cellulose | 22% |
| Carboxymethyl starch is received | 20% |
| Magnesium stearate | 3% |
The above drug and auxiliary material grinding and sieving, mix to obtain the final product.
11 pharmaceutical composition of embodiment
| Compound shown in Formulas I provided by the invention | 15% |
| Starch | 40% |
| Microcrystalline cellulose | 25% |
| Carboxymethyl starch is received | 19% |
| Magnesium stearate | 1% |
The above drug and auxiliary material grinding and sieving, after mixing in tabletting on tablet press machine to get.
11 pharmaceutical composition of embodiment
| Compound shown in Formulas I provided by the invention | 15% |
| Dried starch | 65% |
| Magnesium stearate | 20% |
The above drug and auxiliary material grinding and sieving, be packed into after mixing in hard gelatin capsule to get.
12 pharmaceutical composition of embodiment
| Compound shown in Formulas I provided by the invention | 1% |
| Sodium chloride | 1% |
| Water for injection | 98% |
Compound provided by the invention is added into sodium chloride and appropriate water for injection, is stirred evenly, 0.1% needle activity is added
Decarburization is filtered in carbon, absorption, and benefit injects water to specified amount, micropore filtering film filtering, and by 1mL/ branch encapsulating, 100 DEG C damp and hot to go out
Bacterium 20min, through lamp inspection qualification to get.
13 pharmaceutical composition of embodiment
| Compound shown in Formulas I provided by the invention | 0.1% |
| Ethyl alcohol | 0.4% |
| Sodium chloride | 0.1% |
| Mannitol | 0.6% |
| Water for injection | 99% |
Compound provided by the invention is added into ethyl alcohol stirring and dissolving, adds sodium chloride, mannitol and appropriate water for injection, is stirred
Uniformly, 0.1% needle activated carbon is added, decarburization is filtered in absorption, and benefit injects water to specified amount, and micropore filtering film filtering is pressed
The packing of 4mL/ branch, is freeze-dried, encapsulation, examined it is qualified to get.
Above description is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, it is all the present invention suggested by technical spirit under completed same changes or modifications change, should all belong to
In the covered the scope of the patents of the present invention.
Claims (4)
1. a kind of compound of Formulas I or its pharmaceutically acceptable salt:
2. the synthetic method of compound according to claim 1, which comprises the steps of: sodium hydride to be dissolved in
In n,N-Dimethylformamide, 1- pyridine -6- methoxy-p-carboline and 3- iodobenzene first bromine, Yu Changwen or fire-bar is then added
Under part one pot reaction to get the Formulas I compound；
The reaction carries out under 40 DEG C to organic solvent of reflux temperature；
The synthetic method further includes separation and purification step, and the separation and purifying are that alkali is added dropwise in one pot reaction products therefrom
Property solution be adjusted to neutral or alkalinity, extracted with ethyl acetate or chloroform, collect organic phase, remove solvent, then use silicagel column
The mode of chromatography is purified；
The silica gel column chromatography carries out elution chromatography, the petroleum ether and second using the eluant, eluent that petroleum ether and ethyl acetate form
The volume ratio of acetoacetic ester is 1000:50~100.
3. compound described in claim 1 or its pharmaceutically acceptable salt are preparing anti-kidney fibrosis drug and/or are resisting slow
Application in property nephrosis drug.
4. a kind of pharmaceutical composition, which is characterized in that compound described in claim 1 or its pharmacy including therapeutically effective amount
Upper acceptable salt and pharmaceutically acceptable auxiliary material.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710385798.XA CN107118212B (en) | 2017-05-26 | 2017-05-26 | 1- pyridine -6- methoxyl group -9- (3- iodine benzyl)-B-carboline, synthesis and application |
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