CN105218631A - Imidazopyridine-6-formyl-amino-acid benzyl ester, its synthesis, active and application - Google Patents
Imidazopyridine-6-formyl-amino-acid benzyl ester, its synthesis, active and application Download PDFInfo
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- CN105218631A CN105218631A CN201410271660.3A CN201410271660A CN105218631A CN 105218631 A CN105218631 A CN 105218631A CN 201410271660 A CN201410271660 A CN 201410271660A CN 105218631 A CN105218631 A CN 105218631A
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Abstract
The invention discloses 5 kinds of 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg-AA-OBzl, discloses their preparation method, discloses their restraining effect to tumor growth, disclose their anti-inflammatory activity, also discloses antithrombotic acitivity in their body.
Description
Invention field
The present invention relates to 5 kinds of 3H-imidazo [4,5-c] pyridine-6-formyl-nitro arginyl-amino-acid benzyl esters, relate to their preparation method, relate to their restraining effect to tumor growth, relate to their anti-inflammatory activity, also relate to their antithrombotic acitivity.The invention belongs to biomedicine field.
Background technology
Cancer is one group of common name that can affect the various diseases at any position of health.Other term used is malignant tumour and vegetation.According to World Health Organization's statistics, cancer is one of No.1 cause of the death in the world, especially in developing country.And whole world number of cancer deaths estimates continuation to rise, will more than 1,310 ten thousand to the year two thousand thirty.Therefore, develop new efficient, low toxicity, the antitumor drug that toxic side effect is little is one of important topic of new drug research always.
Along with the understanding to tumor characteristic and morbidity essence, contriver once disclosed 3H-imidazo [4, the 5-c] pyridine of represented by formula
-6
-formyl-AA-OBzl (wherein AA represents sweet amino acid or other L-amino-acid residue) shows good anti-tumor activity under 1 μm of ol/kg dosage.Recognizing by studying contriver further, between the 6-formyl radical and AA-OBzl base of 3H-imidazo [4,5-c] pyridine-6-formyl-AA-OBzl, inserting N
g-NO
2-Arg residue, the metabolism that not only can delay it reaches the object improving anti-tumor activity, and can increase anti-inflammatory and antithrombotic acitivity.Inflammation and thrombus are all the complication of hazard tumor patient prognosis.By this modification, acquisition has antitumor, and the compound of anti-inflammatory and antithrombotic three heavy activity, has good potential applicability in clinical practice.According to these understanding, inventors herein propose the present invention.
Summary of the invention
First content of the present invention is to provide 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg-AA-OBzl (in formula, AA is selected from L-Phe, L-Glu (OBzl), L-Val, L-Pro and L-Met residue).
Second content of the present invention is to provide preparation 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2the synthetic method of-Arg-AA-OBzl (in formula, AA is selected from L-Phe, L-Glu (OBzl), L-Val, L-Pro and L-Met residue), the method comprises:
(1) L-Histidine carries out Pictet-Spengler condensation with formaldehyde and generates 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-carboxylic acid under dilute sulphuric acid catalysis;
(2) 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-Carboxylic Acid is 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-methyl-formiate;
(3) 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-methyl-formiate potassium permanganate oxidation is 3H-imidazo [4,5-c] pyridine-6-methyl-formiate;
(4) 3H-imidazo [4,5-c] pyridine-6-methyl-formiate is saponified into 3H-imidazo [4,5-c] pyridine-6-formic acid in NaOH solution (2N);
(5) 3H-imidazo [4,5-c] pyridine-6-formic acid and N
g-NO
2-Arg-OBzl coupling obtains 3H-imidazo [4,5-c] pyridine-6-formyl N
g-NO
2-Arg-OBzl;
(6) 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg-OBzl is saponified into 3H-imidazo [4,5-c] pyridine-6-formyl-N in NaOH solution (2N)
g-NO
2-Arg;
(7) 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg and L-Phe-OBzl, L-Glu (OBzl)-OBzl, L-Val-OBzl, L-Pro-OBzl and L-Met-OBzl coupling obtain 3H-imidazo [4,5-c] pyridine-6-formyl N
g-NO
2-Arg-AA-OBzl.
3rd content of the present invention evaluates 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg-AA-OBzl is to the restraining effect of tumor cell proliferation.
4th content of the present invention evaluates 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg-AA-OBzl is to the restraining effect of tumor growth.
5th content of the present invention evaluates 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2the anti-inflammatory activity of-Arg-AA-OBzl.
6th content of the present invention evaluates 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2the antithrombotic acitivity of-Arg-AA-OBzl.
Accompanying drawing explanation
Fig. 1 .3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2the synthetic route .i of-Arg-AA-OBzl) HCHO, H
2o, H
2sO
4, 65 DEG C; Ii) MeOH, SOCl
2, 0 DEG C; Iii) DMF, NMM, KMnO
4; Iv) NaOH, H
2o, 0 DEG C; V) dicyclohexylcarbodiimide (DCC), I-hydroxybenzotriazole (HOBt), N-methylmorpholine (NMM), DMF; Vi) NaOH, H
2o, 0 DEG C; Vii) dicyclohexylcarbodiimide (DCC), I-hydroxybenzotriazole (HOBt), N-methylmorpholine (NMM), DMF; AA=L-Phe residue in 8a, AA=L-Met residue in 8b, AA=L-Glu (OBzl) residue in 8c, AA=L-Val residue in 8d, AA=L-Pro residue in 8e.
Embodiment
In order to set forth the present invention further, provide a series of embodiment below.These embodiments are illustrative completely, and they are only used for being specifically described the present invention, not should be understood to limitation of the present invention.
Embodiment 1 prepares 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-carboxylic acid (2)
Under ice bath, 15.0g (96.8mmol) L-Histidine is placed in 250mL round-bottomed flask, add 50mL distilled water, more dropwise add the 3mL vitriol oil, stir, add 15mL formaldehyde solution (40%) after dissolving completely, 60 DEG C are reacted 8 hours.Reactant is cooled to room temperature, lowers pH to 6, have a large amount of colourless precipitate to separate out with strong aqua at ice bath, filters.Filter cake washes with water and drying, and obtaining 15g (93%) title compound, is colorless solid.ESI-MS(m/z)167[M+H]
+。
Embodiment 2 prepares 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-methyl-formiate (3)
In 500mL eggplant bottle, 100mL methyl alcohol is added under ice bath, 10mL thionyl chloride is slowly dripped by constant pressure funnel, 5.0g (30mmol) 6S-4 is added, 5,6 after 1 hour, 7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-carboxylic acid (2), room temperature reaction is after 3 days, and TLC display reacts completely, reaction mixture concentrating under reduced pressure, residue adds dissolve with methanol and concentrating under reduced pressure.This operation repeats 3 times to obtain colourless blister solid, then add diethyl ether drain repetition 3 times colourless powder, finally obtaining 4.2g (55%) title compound with methanol/ether recrystallization, is colorless solid.ESI-MS(m/z)181[M+H]
+。
Embodiment 3 prepares 3H-imidazo [4,5-c] pyridine-6-methyl-formiate (4)
In 100mL eggplant bottle, add 2g (7.9mmol) 6S-4 under ice bath, 5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-methyl-formiate, adds DMF and makes dissolving.In this solution, drip 1mL triethylamine adjusts pH to 8, divides and adds 1.5g (9.4mmol) potassium permanganate for three times.React after 6 hours, TLC monitoring reaction is complete.Reactant is evaporated to dry, the black solid 1NHCl solubilize obtained, and drips 2NNaOH solution and adjusts pH to 7, separate out a large amount of colorless solid under ice bath.This solid is eluent silica column purification with methylene chloride/methanol, obtains 0.93g (66.4%) title compound, is colorless solid.ESI-MS(m/z)177[M+H]
+。
Embodiment 4 prepares 3H-imidazo [4,5-c] pyridine-6-carboxylic acid (5)
In 100mL eggplant bottle, 3mLNaOH solution (1.5N) is added under ice bath, 0.93g (5.3mmol) 3H-imidazo [4 is added after 10min, 5-c] pyridine-6-methyl-formiate (4), react TLC display reaction after 1 hour complete, dripping 2NHCl solution in ice bath downhill reaction liquid adjusts pH to 7, separates out a large amount of colorless solid.This solid is eluent silica column purification with methylene chloride/methanol, obtains 0.56g (65%) title compound, is colorless solid.ESI-MS(m/z)163[M+H]
+。
Embodiment 5 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-OBzl (6)
Take 194mg (1.2mmol) 3H-imidazo [4,5-c] pyridine-6-carboxylic acid in 100mL eggplant-shape bottle, add 20mLDMF.211mg (1.6mmol) HOBt and 333mg (1.6mmol) DCC is added successively, activation 30min at ice bath with under stirring.Take 676mg (1.4mmol) TosN
gnO
2arg-OBzl, in the little triangular flask of 25mL, after dissolving, adjusts pH to 7 with NMM, then drops in the reaction solution of eggplant-shape bottle by this solution with DMF, finally adjusts reacting liquid pH value to 8 with NMM.Room temperature reaction spends the night, and after completion of the reaction, reaction mixture is evaporated to dry in TLC display, and residue adds 50mL methylene dichloride and dissolves, and cross and filter dicyclohexylurea (DCU) (DCU), filtrate layers uses saturated NaHCO successively
3the aqueous solution (20mL × 3) and the saturated NaCl aqueous solution (20mL × 3) respectively wash 3 times, the anhydrous CaCl of ethyl acetate layer
2drying, filters, and filtrate reduced in volume is to dry, the yellow oil obtained is through purification by silica gel column chromatography (methylene chloride/methanol is eluent), the faint yellow solid obtained, obtains 200mg (37%) title compound through methylene dichloride/sherwood oil recrystallization, is colorless solid.ESI-MS (m/e) 454 [M-H]
-dEG C .Mp184-186. [α]
d 25=12.2 (c=0.12, methyl alcohol).
1h-NMR (500MHz, DMSO-d
6) δ/ppm=13.19 (s, 1H), 9.04 (s, 2H), 8.57 (s, 1H), 8.52 (s, 1H), 8.26 (s, 1H), 7.44 (m, 5H), 5.18 (d, J=15.0Hz, 2H), 4.63 (dd, J=5Hz, J=15Hz, 1H), 3.16 (m, 2H), 1.95 (m, 2H), 1.59 (s, 2H).
Embodiment 6 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg (7)
Take 200mg (0.44mmol) 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-OBzl, in 100mL eggplant bottle, adds methanol solution and makes material dissolution, and ice bath stirs the lower 2NNaOH of dropping solution and adjusts pH to 11, and react 3 hours under ice bath, TLC display reaction is complete., drip potassium hydrogen sulfate saturated solution in ice bath downhill reaction liquid and adjust pH to 7, compound of reaction concentrating under reduced pressure, residue methyl alcohol redissolves, elimination insolubles, filtrate reduced in volume, obtains 74mg (46%) title compound, is colorless solid.ESI-MS (m/z): 363 [M-H]
-; Mp166-169 DEG C. [α]
d 25=9.8 (c=0.19, methyl alcohol).
1h-NMR (500MHz, DMSO-d
6) δ/ppm=13.14 (s, 1H), 8.99 (s, 2H), 8.65 (s, 2H), 8.54 (s, 1H), 8.53 (s, 1H), 8.29 (m, 2H), 4.60 (dd, J=5Hz, J=15Hz, 1H), 3.10 (m, 2H), 1.79 (m, 2H), 1.55 (m, 2H).
Embodiment 7 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-Phe-OBzl (8a)
Take 61mg (0.17mmol) 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg, in 100mL eggplant-shape bottle, adds 20mLDMF.27mg (0.2mmol) HOBt and 41.3mg (0.2mmol) DCC is added successively, activation 30min at ice bath with under stirring.Take 73mg (0.17mmol) TosPhe-OBzl in the little triangular flask of 25mL, after dissolving with DMF, adjust pH to 7 with NMM, then this solution is dropped in the reaction solution of eggplant-shape bottle, finally adjust reacting liquid pH value to 8 with NMM.Room temperature reaction spends the night, and TLC display reaction is complete, and reaction mixture is evaporated to dry, and residue adds 20mL methylene dichloride and dissolves, and cross and filter dicyclohexylurea (DCU) (DCU), filtrate layers uses saturated NaHCO successively
3solution (20mL × 3) and saturated NaCl solution (20mL × 3) respectively wash three times, organic over anhydrous CaCl
2dry, filter, filtrate reduced in volume to dry, the yellow oil obtained through purification by silica gel column chromatography (methylene chloride/methanol is eluent), the faint yellow solid obtained, obtaining 25mg (24.7%) title compound through methylene dichloride/sherwood oil recrystallization, is colorless solid.ESI-MS (m/z): 600 [M-H]
-; Mp:147.2-151.8 DEG C; [α]
d 25=-5.3 (c=0.21, methyl alcohol); IR (KBr): 3255,2345,1739,1647,1523,1454,1265,937,698;
1h-NMR (300MHz, DMSO-d6): δ/ppm=13.17 (s, 1H), (8.99 s, 1H), 8.71 (d, J=6Hz, 1H), 8.69 (d, J=6Hz, 1H), 8.56 (s, 2H), 8.25 (s, 1H), (7.87 m, 2H), 7.22 (m, 10H), 5.08 (s, 2H) 4.59 (m, 2H), 3.03 (m, 4H), 1.69 (m, 2H), 1.47 (m, 2H).
Embodiment 8 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-Met-OBzl (8b)
According to the method for embodiment 7, from 182mg (0.5mmol) 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg and 206mg (0.5mmol) TosMet-OBzl obtains 320mg (49%) title compound, is colorless solid.ESI-MS (m/z): 584 [M-H]
-; Mp:126.8-131.7 DEG C; [α]
d 25=-5.3 (c=0.20, methyl alcohol); IR (KBr): 3342,2947,2345,1748,1647,1523,1458,1269,732,698;
1h-NMR (300MHz, DMSO-d6): δ/ppm=13.18 (s, 1H), 9.00 (s, 1H), 8.73 (d, J=9Hz, 1H), 8.65 (d, J=6Hz, 1H), (8.55 s, 2H), 8.25 (s, 1H), (7.92 m, 2H), 7.38 (m, 5H), (5.13 m, 2H), 4.47 (m, 1H), (4.37 m, 1H), 3.15 (m, 2H), 2.45 (m, 2H), (1.96 m, 5H), 1.77 (m, 2H), 1.54 (m, 2H).
Embodiment 9 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-Glu (OBzl)-OBzl (8c)
According to the method for embodiment 7, from 61mg (0.17mmol) 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg and 85mg (0.17mmol) TosGlu (OBzl)-OBzl obtains 34mg (29.8%) title compound, is colorless solid.ESI-MS (m/z): 672 [M-H]
-; Mp:127.2-131.7 DEG C; [α]
d 25=6.2 (c=0.34 methyl alcohol); IR (KBr): 3228,2357,1735,1643,1523,1458,1396,1265,732,628;
1h-NMR (500MHz, DMSO-d6): δ/ppm=13.19 (s, 1H), 8.97 (s, 1H), 8.70 (d, J=8Hz, 1H), 8.61 (d, 7.5=10Hz, 1H), 8.52 (s, 2H), 8.23 (s, 1H), (7.90 m, 2H), 7.31 (m, 10H), 5.06 (m, 4H), (4.60 m, 1H), 4.38 (m, 1H), 3.08 (m, 4H), (2.42 m, 2H), 1.77 (m, 2H), 1.52 (m, 2H).
Embodiment 10 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-Val-OBzl (8d)
According to the method for embodiment 7, from 61mg (0.17mmol) 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg and 60mg (0.17mmol) TosVal-OBzl obtains 21mg (32.7%) title compound, is colorless solid.ESI-MS (m/z): 552 [M-H]
-, Mp:106.2-108.5 DEG C, [α]
d 25=-4.1 (c=0.18, methyl alcohol), IR (KBr): 3387,2962,2376,1732,1643,1523,1454,1269,759,582,
1h-NMR (300MHz, DMSO-d6): δ/ppm=13.18 (s, 1H), 9.00 (s, 1H), 8.75 (d, J=9Hz, 1H), 8.55 (s, 1H), 8.48 (d, J=9Hz, 2H), 8.25 (s, 1H), 7.92 (m, 2H), 7.34 (m, 5H), 5.13 (m, 2H) 4.74 (m, 2H), 4.27 (m, 2H), 3.16 (m, 2H), 2.10 (m, J=6Hz, H), 1.74 (m, 2H), 1.55 (s, 2H), 0.89 (d, J=6Hz, 3H), 0.87 (d, J=6Hz, 3H).
Embodiment 11 prepares 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg-Pro-OBzl (8e)
According to the method for embodiment 7, from 182mg (0.5mmol) 3H-imidazo [4,5-c] pyridine-6-formyl-L-N
g-NO
2-Arg and 189mg (0.5mmol) TosPro-OBzl obtains 114mg (41.5%) colorless solid.ESI-MS (m/z): 550 [M-H]
-dEG C .Mp:133.5-136.2; [α]
d 25=-11.2 (c=0.31, methyl alcohol); IR (KBr): 3224,2951,2360,1739,1627,1523,1450,1334,1257,941,736,698;
1h-NMR (500MHz, DMSO-d6): δ/ppm=13.14 (s, 1H), 8.96 (s, 1H), 8.74 (d, J=7Hz, 1H), 8.53 (s, 2H), 8.23 (s, 2H), 7.81 (m, 2H), 7.33 (m, 6H), 5.11 (m, 2H), 4.80 (m, 1H), 4.42 (m, 1H), 3.70 (m, 2H), (3.12 m, 2H), 2.20 (m, 1H), 1.95 (m, 2H), (1.93 m, 2H), 1.72 (m, 2H), 1.57 (m, 2H).
Experimental example 1 measures the cytotoxicity of compound 8a-e
1) substratum of compound 8a-e of the present invention containing 0.1%DMSO is mixed with desired concn.
2) tumour cell of experiment is MCF-7 (human breast cancer cell), HL60 (people in loop), K562 (human leukemia chronic granulocyte), S180 (mouse ascites oncocyte).
3) experimental technique
MCF-7, HL-60, K562 and S180 cell selects RPMI-1640 substratum; In substratum containing 10% through the foetal calf serum and 1 × 10 of deactivation
5u/L penicillin and 100mg/L Streptomycin sulphate.
The cultivation of attached cell MCF-7 and half attached cell S180: respectively that growth conditions is good, is in the cell of logarithmic phase with 3 × 10
4the density of individual/mL is inoculated in 96 orifice plates, and every hole 100 μ L, is placed in 37 DEG C and 5%CO
2cell incubation case in cultivate 4 hours, then add compound 8a-e through sterilising treatment and the solution be mixed with containing the substratum of 0.1%DMSO by the concentration gradient preset, every hole 25 μ L, control group adds the solvent of isopyknic sample dissolution.Continue cultivation after 48 hours, every hole adds the MTT solution that 25 μ L concentration are 5mg/mL, is placed in 37 DEG C and 5%CO
2cell incubation case in cultivate 4 hours.After careful removing supernatant liquor, every hole adds the DMSO of 100 μ L, and about 10min dissolve purple of vibrating residue (first a ceremonial jade-ladle, used in libation), detects O.D. (absorbancy) value immediately in microplate reader, and wavelength is 570nm.
The cultivation of suspension cell HL60 and K562: respectively that growth conditions is good, is in the cell of logarithmic phase with 5 × 10
4the density of individual/mL is inoculated in 96 orifice plates, every hole 100 μ L, then adds the solution be mixed with the substratum containing 0.1%DMSO through the compound 8a-e of sterilising treatment, every hole 25 μ L by the concentration gradient preset, control group adds the solvent of isopyknic sample dissolution, is placed in 37 DEG C and 5%CO
2cell incubation case in cultivate after 48 hours, every hole adds the MTT solution that 25 μ L concentration are 5mg/mL, and continuing the condition that is placed in is 37 DEG C and 5%CO
2cell incubation case in cultivate 4 hours.The centrifugal 10min of 2500rpm, careful sucking-off supernatant liquor, every hole adds 100 μ LDMSO, and about 10min dissolve purple of vibrating residue (first a ceremonial jade-ladle, used in libation), detects O.D. (absorbancy) value immediately in microplate reader, and wavelength is 570nm.
The activity of compound 8a-e inhibition tumor cell propagation under each concentration is obtained by following formula:
Cell proliferation (%)=(the average O.D. value of compound 8a-e group average O.D. value/control group) × 100%, experiment repetition 3 times, maps to drug level with cell proliferation, obtains IC by graphing method
50(half effective inhibition concentration) value.
4) the results are shown in Table 1.Result shows, compound 8a-e suppresses the IC of four kinds of tumor cell proliferations
50all be greater than 100 μMs, there is no cytotoxicity.
Extracorporeal anti-tumor cell-proliferation activity (the IC of table 1 compound 8a-e
50, μM)
The activity of experimental example 2 assessing compound 8a-e Tumor suppression growth
1) physiological saline solution of compound 8a-e tween 80 of the present invention, Zorubicin physiological saline solution is as positive control, and the physiological saline of tween 80 is as negative control;
2) compound 8a-e, physiological saline and the Zorubicin of tween 80 are intraperitoneal administration, the dosage of compound 8a-e is 100nmol/kg and 20nmol/kg, the dosage of the physiological saline of tween 80 is 0.2mL/20g, Zorubicin dosage is 2 μm of ol/kg, successive administration 7 days, altogether administration 7 times.
3) laboratory animal is ICR male mice (cleaning grade), body weight 20 ± 2g, often organizes 12 mouse.
4) knurl source is mouse S 180 sarcoma, purchased from Department Of Medicine, Peking University's animal experimental center, and maintenance of going down to posterity voluntarily.
5) animal model and treatment
The eugonic S180 ascitic tumor knurl liquid of inoculation is extracted under aseptic condition, the liquid of (1: 2) is become fully to mix with normal saline dilution, by freshly prepared 0.2% Trypan Blue of tumor cell suspension, by white blood cell count(WBC) method counting after mixing, contaminate blue person for dead cell, tinter is not viable cell, and is calculated as follows cell concn and cell survival rate.
Viable count/4 × 10 in the block plaid of cell concn=4
4× extension rate=cell count/mL
Cell survival rate=viable count/(viable count+dead cell number) × 100%
Knurl liquid homogenate method survival rate being greater than 90% is prepared into 2.0 × 10
7the cell suspension of individual/mL, in the subcutaneous vaccination of mouse armpit, 0.2mL/ only, manufactures S180 tumor-bearing mice.After tumor inoculation 24h, treatment group mouse abdominal injection every day compound 8a-e, dosage is 100nmol/kg and 20nmol/kg.The physiological saline of naive mice abdominal injection every day 0.2mL tween 80.Positive controls mouse abdominal injection every day Zorubicin, dosage is 2 μm of ol/kg.Experiment proceeds to the 8th day, claim Mouse Weight, etherization, de-cervical vertebra puts to death mouse, then fixes the right armpit tumor location of mouse with tweezers, cuts off skin, expose tumour, blunt separation, weighs, and is calculated as follows tumour inhibiting rate: the average knurl of tumour inhibiting rate %=(negative control group average knurl weight-compound group average knurl weight)/negative control group heavy × 100%.Experimental data adopts t inspection and variance analysis, knurl heavy with
represent.The results are shown in Table 2.As can be seen from Table 2, under 100nmol/kg dosage, compound 8a, the knurl of b, d, e treatment mouse is heavily significantly less than the knurl weight of the saline-treated mice of tween 80, and compound 8a is described, the effective dose of b, d, e is low reaches 100nmol/kg.This effective dose is lower 10 times than effective dose 1 μm of ol/kg of contriver once disclosed 3H-imidazo [4,5-c] pyridine-6-formyl-AA-OBzl (wherein AA represents sweet amino acid or other L-amino-acid residue).
Be it can also be seen that by table 2, be heavily significantly less than the knurl weight of the saline-treated mice of tween 80 in the knurl of 20nmol/kg dosages for Compound 8e treatment mouse, illustrate that the effective dose of compound 8e is low and reach 20nmol/kg.This effective dose is lower 50 times than effective dose 1 μm of ol/kg of contriver once disclosed 3H-imidazo [4,5-c] pyridine-6-formyl-AA-OBzl (wherein AA represents sweet amino acid or other L-amino-acid residue).
Table 2 compound 8a-e is on the impact of S180 tumor-bearing mice tumor growth
N=15; Physiological saline is the physiological saline of tween 80; A) with physiological saline group than p < 0.01; B) with physiological saline group than p < 0.01, with 20nmol/kg8 group than p < 0.05.
The anti-inflammatory activity of experimental example 3 assessing compound 8a-e
1) experimental technique
18-22gICR male mice is divided into blank group, positive medication group and administration group at random, and tranquillization 1 day before mouse uses, operation room keeps room temp 22 DEG C, often organizes mouse 10.The left ear gabarit of single administration toward small white mouse after 30 minutes is coated with dimethylbenzene (0.03mL), is put to death by small white mouse cervical dislocation after 2 hours.By a left side for mouse, auris dextra is cut, and with the punch tool of diameter 7mm in the same position of two ears, gets circular auricle, weighs respectively, obtains the weight difference of two circle auricles as swelling.Swelling=left ear disk weight-auris dextra disk weight.
2) medication and dosage
Administering mode is gastric infusion.Blank: physiological saline, dosage is 0.2mL/20g; Positive control: acetylsalicylic acid, dosage is 1.11mmol/kg; The dosage of the compounds of this invention 8a-e is 20nmol/kg.
3) statistical method
This experimental data statistics all adopt t inspection and variance analysis, swelling with
represent.
4) experimental result lists table 3 in.
Result shows, is starkly lower than the ear swelling degree of the saline-treated mice of tween 80 at the ear swelling degree of 20nmol/kg dosages for Compound 8a, e treatment mouse.Compound 8a is described, e, under the dosage playing Anticancer effect in vivo, also has anti-inflammatory action.
The interior anti-inflammatory activity evaluation of table 3 compound 8a-e
N=10; Physiological saline is the physiological saline of tween 80; A) with the physiological saline group of tween 80 than p < 0.01.
Antithrombotic acitivity in the body of experimental example 4 assessing compound 8a-e
1) experiment material
Laboratory animal: SD male rat (cleaning grade), body weight 200 ± 20g, purchased from Beijing Vital River Experimental Animals Technology Co., Ltd., every 12 rats are one group, blank and each one group of positive control;
Experiment intubate is formed by 3 sections, middle segment length 8.0cm, internal diameter 0.3cm, two ends are identical polyethylene tube, long 10.0em, internal diameter 0.1cm, external diameter 0.2cm, one end of this pipe pulls into point pipe (for inserting rat carotid artery or jugular vein), all uses the silicon ether silanization of 1% before the inwall use of 3 sections of pipes.The silk thread of the long 6.0cm weighed in advance is put into stage casing polyethylene extra heavy pipe, and the two ends of extra heavy pipe are nested with the non-drawing-down end of two polyethylene tubules respectively (wherein one section silk thread is pushed down 0.5mm fix).For subsequent use by filling heparin-saline solution (50IU/kg) in pipe by sharp pipe end with syringe.
2) foundation of animal model
After gastric infusion 30min or abdominal injection, by rat with 20% urethane solution (6mL/kg, i.p.) anaesthetize.Anesthetized rat dorsal position is fixed, isolate the left side external jugular vein of rat, proximal part and distal end penetrate surgical thread respectively, ligation distal end, the left external jugular vein exposed cuts an angle carefully, the non-line ball end point pipe of the intubate prepared is inserted the proximal part of left external jugular vein opening by angle, pushed the heparin-saline (50IU/kg) of correct amount by the sharp pipe of the other end with syringe, now syringe does not withdraw polyethylene tube, be separated right carotid, in proximal part folder bulldog clamp, proximal part and distal end penetrate surgical thread respectively, ligation distal end, right common carotid artery is being cut an angle carefully nearby from bulldog clamp.Extract syringe from the tip of polyethylene tube, the tip of polyethylene tube is inserted the proximal part of artery angle.The two ends of bypass duct all use 4 trumpeter's art suture ligatures to fix.Open bulldog clamp, make blood flow flow to jugular vein by bypass duct from carotid artery.From circulation time timing, take out from bypass duct after 15min and hang with the silk thread of thrombus, accurately weigh, of poor quality before and after silk thread is wet weight of thrombus.
3) statistical method
This experimental data statistics all adopts t inspection and variance analysis, bolt heavy with
represent.
4) medication and dosage
Administering mode is gastric infusion.Blank: physiological saline, dosage is 0.6mL/200g; Positive control: acetylsalicylic acid, dosage is 167 μm of ol/kg; The dosage of the compounds of this invention 8a-e is 20nmol/kg.
5) experimental result is in table 4.
As can be seen from Table 4, the thrombus weight of compound 8b, c treatment rat is significantly less than the thrombus weight of the saline therapy rat of tween 80.Show compound 8b, c, under the dosage playing Anticancer effect in vivo, also has the activity that anti-arterial thrombus generates.
Anti-bolt evaluation in the body of table 4 compound 8a-e
N=12; A) p < 0.01 is compared with physiological saline group.
Claims (5)
1. 5 kinds of 3H-imidazo [4,5-c] pyridine-6-formyl-N of following formula
g-NO
2-Arg-AA-OBzl (in formula, AA is selected from L-Phe, L-Glu (OBzl), L-Val, L-Pro and L-Met residue).
2. 3H-imidazo [4, the 5-c] pyridine-6-formyl-N of claim 1
g-NO
2the preparation method of-Arg-AA-OBzl (in formula, AA is selected from L-Phe, L-Glu (OBzl), L-Val, L-Pro and L-Met residue), the method comprises:
(1) L-Histidine carries out Pictet-Spengler condensation with formaldehyde and generates 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-carboxylic acid under dilute sulphuric acid catalysis;
(2) 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-Carboxylic Acid is 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-methyl-formiate;
(3) 6S-4,5,6,7-tetrahydrochysene-3H-imidazo [4,5-c] pyridine-6-methyl-formiate potassium permanganate oxidation is 3H-imidazo [4,5-c] pyridine-6-methyl-formiate;
(4) 3H-imidazo [4,5-c] pyridine-6-methyl-formiate is saponified into 3H-imidazo [4,5-c] pyridine-6-formic acid in NaOH solution (2N);
(5) 3H-imidazo [4,5-c] pyridine-6-formic acid and N
g-NO
2-Arg-OBzl coupling obtains 3H-imidazo [4,5-c] pyridine-6-formyl N
g-NO
2-Arg-OBzl;
(6) 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg-OBzl is saponified into 3H-imidazo [4,5-c] pyridine-6-formyl-N in NaOH solution (2N)
g-NO
2-Arg;
(7) 3H-imidazo [4,5-c] pyridine-6-formyl-N
g-NO
2-Arg and L-Phe-OBzl, L-Glu (OBzl)-OBzl, L-Val-OBzl, L-Pro-OBzl and L-Met-OBzl coupling obtain 3H-imidazo [4,5-c] pyridine-6-formyl N
g-NO
2-Arg-AA-OBzl.
3. 3H-imidazo [4, the 5-c] pyridine-6-formyl-N of claim 1
g-NO
2-Arg-AA-OBzl is preparing the application in antitumor drug.
4. 3H-imidazo [4, the 5-c] pyridine-6-formyl-N of claim 1
g-NO
2-Arg-AA-OBzl is preparing the application in anti-inflammatory drug.
5. 3H-imidazo [4, the 5-c] pyridine-6-formyl-N of claim 1
g-NO
2-Arg-AA-OBzl is preparing the application in antithrombotic reagent.
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|---|---|---|---|---|
| US4812462A (en) * | 1986-04-01 | 1989-03-14 | Warner-Lambert Company | 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid analogs having antihypertensive activity |
| CN102796165A (en) * | 2011-05-24 | 2012-11-28 | 首都医科大学 | Arg-Tyr modified aliphatic amine/ alcohol derivatives and preparation method and application thereof |
| CN103450329A (en) * | 2012-05-29 | 2013-12-18 | 首都医科大学 | 3H-imidazo[4,5-c]pyridine-6-formyl-amido acid benzyl esters and their synthesis, anti-tumor activity and use |
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2014
- 2014-06-13 CN CN201410271660.3A patent/CN105218631A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4812462A (en) * | 1986-04-01 | 1989-03-14 | Warner-Lambert Company | 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid analogs having antihypertensive activity |
| CN102796165A (en) * | 2011-05-24 | 2012-11-28 | 首都医科大学 | Arg-Tyr modified aliphatic amine/ alcohol derivatives and preparation method and application thereof |
| CN103450329A (en) * | 2012-05-29 | 2013-12-18 | 首都医科大学 | 3H-imidazo[4,5-c]pyridine-6-formyl-amido acid benzyl esters and their synthesis, anti-tumor activity and use |
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