CN104610424B - Lys (Pro-Ala-Lys) curcumin derivate, its synthesis and the application in medicine - Google Patents
Lys (Pro-Ala-Lys) curcumin derivate, its synthesis and the application in medicine Download PDFInfo
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Abstract
The invention discloses following formula puppet peptides, and the present invention further discloses their preparation method and purposes.The pseudo- peptide of the present invention has outstanding thrombus dissolving activity, antithrombotic acitivity, anti-inflammatory activity, OH free radical scavenging activities and nanostructure.
Description
The application be filed in 14 days December in 2011 application No. is：201110415171.7 it is entitled " Lys and
The divisional application of the patent of the curcumin derivate of Lys (Pro-Ala-Lys) modifications, its synthesis and the application in medicine ".
Technical field
The present invention relates to the pseudo- peptides of following formula, and the invention further relates to their preparation method and purposes.The present invention
Compound remove have outstanding thrombus dissolving activity, antithrombotic acitivity, anti-inflammatory activity, OH free radical scavenging activities and nano junction
Structure.The invention belongs to biomedicine fields.
Background technology
Curcumin is one of active ingredient of turmeric, has antithrombotic, anti-inflammatory and free-radical scavenging activity, is safety and low toxicity
Natural products.Inflammatory reaction promotes release and the platelet activation of coagulation factor, while lowering natural anticoagulant and inhibiting fine
Molten activity, thrombosis attend by the process that acute inflammation is converted to unrestrained property inflammation.The inflammation of appropriateness is conducive to by various
The body of damage maintains the stabilization of environment in itself, is conducive to the recovery that body safeguards the complete and function of self structure.However,
Too strong inflammatory reaction may aggravate the damage of body, generate the chain reaction of a sequence, cause vicious circle.In thromboembolism treatment
In give anti-inflammatory drug, help to improve thrombolysis rate, mitigate tissue damage.Ischemical reperfusion injury is that damage is organized after causing thrombolysis
One reason of wound, free radical is a key factor for causing reperfusion injury.Radicals scavenging is given in thromboembolism treatment
Agent has protective effect for ischemical reperfusion injury.Although curcumin platelet aggregation-against model, external free radical in vitro
It removes and shows the work expected on model, rat arteriovenous bypass intubation antithrombotic model and mice ear anti-inflammation models
Property, and these activity have protection to make cerebral ischemia re-pouring injured, myocardial ischemia-reperfusion injury, lung reperfusion injury
With, but curcumin does not have thrombus dissolving activity, that is, the premise for generating ischemia-reperfusion to be not present.It was recognized by the inventor that only past
Thrombus dissolving oligopeptides is introduced in curcumin could realize its protective effect.
P6A (Ala-Arg-Pro-Ala-Lys) is one of fibrin β chain degradation products, has thrombus dissolving activity.Invention
People is found that metabolite PAK in the metabolism research of P6A.It is bypassed on intubation thrombus dissolving model in rat arteriovenous, PAK's is molten
Thrombus activity is stronger than parent P6A.According to general understanding, polypeptide can all degrade rapidly in vivo.Prolonged by the structural modification of PAK
Delay internal degradation rate and improve thrombus dissolving activity, is the important channel of oligopeptides thrombolytic agent research.
According to general understanding, the amphipathic molecule containing polypeptide, such as aliphatic alcohol chain modification polypeptide, in condition appropriate
Down nanostructure can be formed by noncovalent intermolecular interactions.It can improve polypeptide in vivo defeated by nanostructure
Send, delay the activity in vivo of the degradation rate and raising polypeptide of polypeptide in vivo.According to these understanding, this hair is inventors herein proposed
It is bright.
Invention content
First content of the present invention is the pseudo- peptide of formula
Second content of the present invention is the method for preparing above formula puppet peptide, and this method is made of following steps：
(1) in the presence of dicyclohexylcarbodiimide (DCC) and n-hydroxysuccinimide (HOSu) Boc-Pro anhydrous
It is Boc-Pro-OSu to be condensed with HOSu in THF, in NaHCO3In the presence of Boc-Pro-OSu reacted with L-Ala generate Boc-Pro-
Ala；
(2) Boc-Pro-Ala is Boc- with Lys (Boc)-OBzl condensations in anhydrous THF in the presence of DCC and HOBt
Pro-Ala-Lys(Boc)-OBzl；
(3) Boc-Pro-Ala-Lys (Boc)-OBzl is hydrolyzed to Boc-Pro-Ala- in methyl alcohol in the presence of NaOH
Lys(Boc)；
(4) in anhydrous K2CO3In the presence of in anhydrous THF vanillic aldehyde reacted with bromoacetate generate 3- methoxyl group -4- oxygen
Acetyl triethyl benzaldehyde；
(5) in B2O3、(nBuo)3In the presence of B and nBu-NH Veratraldehyde in anhydrous ethyl acetate with second
Acyl condensation of acetone is 6- (3,4- bis--anisyl) -5,6- hexene -2,4- diketone；
(6) in B2O3、(nBuo)3In the presence of B and nBu-NH, the 6- (4- methyl -3- methoxybenzenes in anhydrous ethyl acetate
Base) condensation of -5,6- hexenes -2,4- diketone and 3- methoxyl group -4- oxygen acetyl triethyl benzaldehydes is 1- (4- methyl -3- methoxyl groups
Phenyl) -7- (4- oxygen acetyl triethyl base -3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(7) in the presence of NaOH, in acetone by 1- (4- methyl -3- methoxyphenyls) -7- (4- oxygen acetyl triethyls bases -
3- methoxyphenyls) -1,6- heptadiene -3,5- diketone is hydrolyzed to 1- (4- methyl -3- methoxyphenyls) -7- (4- fluoroacetic acid bases -
3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(8) in the presence of DCC and HOBt in anhydrous THF 1- (4- methyl -3- methoxyphenyls) -7- (4- fluoroacetic acid bases -
3- methoxyphenyls) -1,6- heptadiene -3,5- diketone and Lys (Boc)-OBzl condensation is 1- (4- methyl -3- methoxybenzenes
Base) -7- (4- oxygen acetyl-Lys (Boc)-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(9) 1- (4- methyl -3- methoxyphenyls) -7- (the 4- oxygen acetyl-Lys in hydrogen chloride-ethyl acetate solution
(Boc)-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone slough Boc generate 1- (4- methyl -3- methoxybenzenes
Base) -7- (4- oxygen acetyl-Lys-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(10) in the presence of DCC and HOBt in anhydrous THF Boc-Pro-Ala-Lys (Boc) and 1- (4- methyl -3- first
Phenyl) -7- (4- oxygen acetyl-Lys-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone condensation be 1- (4- first
Base -3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,
6- heptadiene -3,5- diketone；
(11) in methyl alcohol by 1- (4- methyl -3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys in the presence of NaOH
[Boc-Pro-Ala-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone is hydrolyzed to 1- (4- methyl -
3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)] -3- methoxyphenyls } -1,6- heptadiene -
3,5- diketone；
(12) by 1- (4- methyl -3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys in hydrogen chloride-ethyl acetate solution
[Boc-Pro-Ala-Lys (Boc)] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone slough Boc generate 1- (4- methyl -
3- methoxyphenyls) -7- [4- oxygen acetyl-Lys (Pro-Ala-Lys) -3- methoxyphenyls] -1,6- heptadiene -3,5- diketone.
The third content of the present invention is the thrombus dissolving activity of the pseudo- peptide of evaluation.
The 4th content of the present invention is the antithrombotic acitivity of the pseudo- peptide of evaluation.
The 5th content of the present invention is the anti-inflammatory activity of the pseudo- peptide of evaluation.
The 6th content of the present invention is the OH free radical scavenging activities of the pseudo- peptide of evaluation.
The 7th content of the present invention is the nanostructure of the pseudo- peptide of characterization.
Description of the drawings
Fig. 1 synthetic route .3a-7a R=H, 3b-7b R=CH3。
The transmission electron microscope photo of Fig. 2 compound 5a nano-rings.
The transmission electron microscope photo of Fig. 3 compound 5b nanotubes.
The nanosphere transmission electron microscope photo of Fig. 4 compounds 7a.
The nanometer rods transmission electron microscope photo of Fig. 5 compounds 7b.
Specific implementation mode
In order to which the present invention is further explained, a series of embodiments are given below.These embodiments be entirely it is illustrative, it
Only be used for the present invention is specifically described, be not construed as limitation of the present invention.
Embodiment 1 prepares Boc-Pro-Ala-Lys (Boc)
1) preparation of Boc-Pro-Ala
1.075g (5.0mmol) Boc-Pro is dissolved in the anhydrous THF of 20mL, 0.637g is added into solution under ice bath
(5mmol) n-hydroxysuccinimide (HOSu), and make to be completely dissolved.Under ice bath, two hexamethylenes for having been dissolved in a small amount of anhydrous THF
Base carbonyl diimine (DCC) 1.236g (6.0mmol) is added in reaction solution.It is stirred at room temperature 7h, TLC (petrol ether/ethyl acetate, 3:
1) monitoring Boc-Pro disappears.Dicyclohexylurea (DCU) (DCU) is filtered out, filtrate decompression concentration removes THF.Residue with Ethyl acetate is molten
Solution, solution is successively with saturation NaHCO3Aqueous solution, which washes, is saturated NaCl aqueous solutions washes, and is then concentrated under reduced pressure into ethyl acetate layer
The aqueous solution that appropriate THF dissolvings add 0.489g (5.5mmol) Ala later is added in dry, residue.Mixture NaHCO3Gu
Body tune pH 9 reacts at room temperature 12h, and reaction mixture, which is concentrated under reduced pressure, removes THF, and residue adds 5mL water dissolutions, aqueous solution saturation
KHSO4Aqueous solution tune pH 2 is extracted 5 times with 30mL ethyl acetate, and combined ethyl acetate layer is washed till with saturation NaCl aqueous solutions
Neutrality, anhydrous sodium sulfate drying.Filtering, filtrate decompression are concentrated to dryness, and obtain 1.35g (94%) title compound, are colorless solid.
ESI-MS(m/e)：285[M-H]-。
2) preparation of Boc-Pro-Ala-Lys (Boc)-OBzl
1.202g (4.2mmol) Boc-Pro-Ala is dissolved in the anhydrous THF of 20mL, 0.544g is added into solution under ice bath
(4mmol) HOBt is added 1.030g (5mmol) DCC after being completely dissolved, obtains reaction solution (I), stirs 30 minutes.
2.168g (4.0mmol) Lys (Boc)-OBzl is suspended in the anhydrous THF of 20mL, and 1mLN- methyl morpholines (NMM) are then added, and is adjusted
PH 9 obtains reaction solution (II).Reaction solution (II) is added in reaction solution (I) after stirring 1h under ice bath, 4h is stirred at room temperature,
TLC (chloroform/methanol, 15: 1) showing that Lys (Boc)-OBzl disappears.DCU is filtered out, filtrate decompression concentration removes THF.Residue is used
50mL ethyl acetate dissolves.Obtained solution is successively with saturation NaHCO3Aqueous solution, which washes, is saturated NaCl aqueous solutions washes, 5%KHSO4
Aqueous solution is washed and is saturated NaCl aqueous solutions and washes.Ethyl acetate layer anhydrous Na2SO4Dry, filtering, filtrate decompression is concentrated to dryness, obtains
It is colourless powder to 1.915g (75%) title compound.ESI-MS(m/e)：604[M+H]+。
3) preparation of Boc-Pro-Ala-Lys (Boc)
1.585g (3.0mmol) Boc-Pro-Ala-Lys (Boc)-OBzl is dissolved in 15mL methanol.It will be obtained under ice bath
Solution is with NaOH (2N) aqueous solution tune pH12 and stirs 2h, and TLC (dichloro/methanol, 15: 1) showing Boc-Pro-Ala-Lys
(Boc)-OBzl disappears.Reaction mixture 2N dilute hydrochloric acid tune pH 2 is concentrated under reduced pressure and removes methanol.Residue with Ethyl acetate extracts
3 times.Combined ethyl acetate layer is washed till neutrality, anhydrous Na with saturation NaCl aqueous solutions2SO4It is dry.Filtering, filtrate decompression concentration
To doing, 1.397g (85%) title compound is obtained, is colorless solid.ESI-MS(m/e)：514[M-H]-。
Embodiment 2 prepares 1- (3- methoxyl group -4- hydroxyphenyls) -7- [(4- fluoroacetic acid base) -3- methoxyphenyls] -1,6- heptan
Diene -3,5- diketone
1) preparation of 6- (3- methoxyl group -4- hydroxyphenyls) -5,6- hexene -2,4- diketone
31mL (0.3mol) acetylacetone,2,4-pentanedione is placed in 250mL three-necked bottles, be added 14.5g (0.21mol) diboron trioxides and
70mL ethyl acetate, 70 DEG C of reactions make to become white suspension for one hour, and three positive fourth of 29.5mL (0.11mol) boric acid is then added
Ester and 15.2g (0.1mol) 3- methoxy -4- hydroxy benzaldehydes, 85 DEG C of reactions make within 0.5 hour solution shoal yellow suspension, pass through
The ethyl acetate solution of 10.88mL (0.11mol) n-butylamine is added dropwise in constant pressure funnel, and reaction makes solution become red for 1 hour, cooling
To 50 DEG C, 200mL 1N hydrochloric acid be added, reaction 0.5 hour, stop reaction.Separate water layer, with 40mL ethyl acetate extraction three times,
Combined ethyl acetate, with anhydrous sodium sulfate it is dry, filter out sodium sulphate, filtrate decompression removes solvent, residue silica gel column chromatography
It purifies, obtain 8.2g (34.6%) title compound, be faint yellow solid.ESI-MS(m/e)：235[M+H]+。
2) preparation of 3- methoxyl groups -4- oxygen acetyl triethyl benzaldehyde
7.6g (50mmol) vanillic aldehyde is placed in 250mL eggplant bottles and is dissolved with anhydrous THF, 9.66g is then added
(70mmol) Anhydrous potassium carbonate, reaction 30min make mixture become white opacity liquid.5.93mL (55mmol) bromoacetic acid second is added dropwise
Ester, reaction 48h, TLC show that vanillic aldehyde disappears.Filter out that insoluble matter, that solvent, residue 200mL ethyl acetate is removed under reduced pressure is molten
Solution, with saturated sodium bicarbonate aqueous solution wash three times, be added anhydrous sodium sulfate drying.11.316g is obtained after removing solvent
(95.1%) title compound is colorless solid.ESI-MS(m/e)：239[M+H]+。
3) 1- (3- methoxyl group -4- hydroxyphenyls) -7- (4- oxygen acetyl triethyl -3- methoxyphenyls) -1,6- heptadiene -3,5-
The preparation of diketone
By in embodiment 2 1) preparation method of item by 1.097g (4.42mmol) 6- (3- methoxyl group -4- hydroxyphenyls) -5,6-
Hexene -2,4- diketone and the reaction of 1.088g (4.87mmol) 3- methoxyl group -4- oxygen acetyl triethyl benzaldehydes, through silica gel column chromatography
0.435g (21.0%) title compound is obtained after purification, is faint yellow solid.ESI-MS(m/e)：455[M+H]+。
4) 1- (3- methoxyl group -4- hydroxyphenyls) -7- (4- fluoroacetic acid base -3- methoxyphenyls) -1,6- heptadiene -3,5- bis-
The preparation of ketone is by 454mg (1mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- (4- oxygen acetyl triethyl base -3- methoxyphenyls) -
1,6- heptadiene -3,5- diketone is placed in 100mL eggplant bottles, addition acetone makes dissolving, solution yellow transparent.Then 1mL 4N are added dropwise
Sodium hydrate aqueous solution, solution gradually become reddish black from red.After 5min, there is yellow solid precipitation.After 10min, TLC is shown
1- (3- methoxyl group -4- hydroxyphenyls) -7- (4- oxygen acetyl triethyl base -3- methoxyphenyls) -1,6- heptadiene -3,5- diketone disappears
It loses.2N HCl/water solution is added dropwise, adjusts pH value to neutrality, is removed under reduced pressure acetone, residue filtering, precipitated with originally water washing,
424mg (96.4%) title compound is obtained, is orange powder.ESI-MS(m/e)：439[M-H]-。
Embodiment 3 prepares 1- (3,4- bis--anisyl) -7- [(4- fluoroacetic acid base) -3- methoxyphenyls] -1,6- heptan two
Alkene -3,5- diketone
1) preparation of 6- (3,4- bis--anisyl) -5,6- hexene -2,4- diketone
By in embodiment 2 1) preparation method of item by 16.6g bis--methoxybenzaldehydes of (0.1mol) 3,4- and 31mL
(0.3mol) acetylacetone,2,4-pentanedione reacts, and purifies to obtain 8.6g (34.6%) title compound through column chromatography, is faint yellow solid.ESI-
MS(m/e)：235[M+H]+。
2) 1- (3,4- bis--anisyl) -7- (4- oxygen acetyl triethyl -3- methoxyphenyls) -1,6- heptadiene -3,5- bis-
The preparation of ketone
By in embodiment 2 1) preparation method of item by 1.097g (4.42mmol) 6- (3,4- bis--anisyl) -5,6- oneself
Alkene -2,4- diketone and the reaction of 1.088g (4.87mmol) 3- methoxyl group -4- oxygen acetyl triethyl benzaldehydes, it is pure through silica gel column chromatography
0.435g (21.0%) title compound is obtained after change, is faint yellow solid.ESI-MS(m/e)：469.6[M+H]+。
3) 1- (3,4- bis--anisyl) -7- (4- fluoroacetic acid base -3- methoxyphenyls) -1,6- heptadiene -3,5- diketone
System it is each
By in embodiment 2 4) preparation method of item by 454mg (1mmol) 1- (3,4- bis--anisyl) -7- (4- oxygen second
Acyl ethyl ester -3- methoxyphenyls) -1,6- heptadiene -3,5- diketone hydrolyzes to obtain 447mg (98%) title compound, and it is orange powder
End.ESI-MS(m/e)：454[M-H]-。
Embodiment 4 prepares 1- (3- methoxyl group -4- hydroxyphenyls) -7- [(4- oxygen acetyl-Lys) -3- methoxyphenyls] -1,6-
Heptadiene -3,5- diketone (5a)
1) 1- (3- methoxyl group -4- hydroxyphenyls) -7- { [4- oxygen acetyl-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,
The preparation of 6- heptadiene -3,5- diketone
By in embodiment 1 2) preparation method of item by 4.4g (10mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- (4- oxygen
Acetate -3- methoxyphenyls) -1,6- heptadiene -3,5- diketone and 5.08g (10mmol) Lys (Boc)-OBzl reaction, obtain
7.02g (94%) title compound is yellow powder.ESI-MS(m/e)：745[M+H]+.1H NMR (300MHz, CDCl3-
d1)：δ/ppm=7.65-7.58 (m, 2H), 7.45 (d, J=7.2Hz, 1H), 7.38-7.33 (m, 5H), 7.16-7.07 (m,
4H), 6.97-6.90 (m, 2H), 6.57-6.48 (m, 2H), 5.84 (s, 1H), 5.24-5.15 (m, 2H), 4.77-4.73 (m,
1H), 4.71-4.56 (m, 2H), 4.50 (bs, 0.91H), 3.97 (s, 3H), 3.94 (s, 3H), 3.06-3.04 (m, 2H),
1.96-1.66 (m, 2H), 1.45 (s, 11H), 1.33 (m, 2H).
2) 1- (3- methoxyl group -4- hydroxyphenyls) -7- [(4- oxygen acetyl-Lys-OBzl) -3- methoxyphenyls] -1,6- heptan two
The preparation of alkene -3,5- diketone
By 3.72g (5mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- { [4- oxygen acetyl-Lys (Boc)-OBzl] -3- first
Phenyl } -1,6- heptadiene -3,5- diketone is dissolved in hydrogen chloride-ethyl acetate solution of 40mL 4N, and ice bath stirring 2 is small
When, TLC (dichloro/methanol, 15/1) shows 1- (3- methoxyl group -4- hydroxyphenyls) -7- (4- oxygen acetyl-Lys (Boc)-OBzl-3-
Methoxyphenyl) disappearance of -1,6- heptadiene -3,5- diketone.It is concentrated under reduced pressure and removes ethyl acetate, residue adds a small amount of ether repeatedly
It is concentrated under reduced pressure to remove hydrogen chloride gas, finally obtains 3.32g (98%) title compound, be yellow powder.ESI-MS(m/
e)：645[M+H]+。
3) 1- (3- methoxyl group -4- hydroxyphenyls) -7- [(4- oxygen acetyl-Lys) -3- methoxyphenyls]-heptadiene -3 1,6-,
The preparation of 5- diketone
By in embodiment 1 3) preparation method of item by 2.57g (4mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- [(4- oxygen
Acetyl-Lys-OBzl) -3- methoxyphenyls] -1,6- heptadiene -3,5- diketone hydrolyzes to obtain 2.15g (93%) title compound,
For yellow powder.ESI-MS(m/e)：555[M+H]+.1H NMR (300MHz, DMSO-d6)：δ/ppm=8.42-8.40 (m,
1H), 8.19 (bs, 0.7H), 7.59 (s, 1H), 7.54 (s, 1H), 7.39-7.14 (m, 4H), 6.97-6.76 (m, 4H), 6.12
(s, 1H), 4.64 (s, 2H), 4.25 (m, 1H), 3.86 (s, 3H), 3.83 (s, 3H), 2.73-2.71 (m, 2H), 1.74 (m,
2H), 1.54 (m, 2H), 1.33 (m, 2H).
Embodiment 5 prepares 1- (3,4- bis--anisyl) -7- [(4- oxygen acetyl-Lys) -3- methoxyphenyls] -1,6- heptan
Diene -3,5- diketone (5b)
1) 1- (3,4- bis--anisyl) -7- { [4- oxygen acetyl-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,6-
The preparation of heptadiene -3,5- diketone
By in embodiment 1 2) each method processed of item by 4.6g (10mmol) 1- (3,4- bis--anisyl) -7- [(4- oxygen second
Acidic group) -3- methoxyphenyls] -1,6- heptadiene -3,5- diketone and 5.08g (10mmol) Lys (Boc)-OBzl reaction, obtain
7.24g (94%) title compound is yellow powder.ESI-MS(m/e)：759[M+H]+.1H NMR (300MHz, CDCl3-
d1)：δ/ppm=7.66-7.580 (m, 2H), 7.45 (d, J=7.2Hz, 1H), 7.38-7.33 (m, 5H), 7.17-7.10 (m,
4H), 6.92-6.88 (m, 2H), 6.56-6.50 (m, 2H), 5.85 (s, 1H), 5.23-5.14 (m, 2H), 4.74-4.72 (m,
1H), 4.66-4.60 (m, 2H), 4.55 (bs, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 3.93 (s, 3H), 3.03 (m, 2H),
1.91-1.73 (m, 2H), 1.46 (s, 11H), 1.30 (m, 2H).
2) 1- (3,4- bis--anisyl) -7- [(4- oxygen acetyl-Lys-OBzl) -3- methoxyphenyls] -1,6- heptan two
The preparation of alkene -3,5- diketone
By in embodiment 4 2) preparation method of item by 3.81g (5mmol) 1- (3,4- bis--anisyl) -7- { [(4- oxygen
Acetyl-Lys (Boc)-OBzl) -3- methoxyphenyls -1,6- heptadiene -3,5- diketone removing Boc obtain 3.45g (98%) title
Compound is yellow powder.ESI-MS(m/e)：660[M+H]+。
3) 1- (3,4- bis--anisyl) -7- [(4- oxygen acetyl-Lys) -3- methoxyphenyls] -1,6- heptadiene -3,5-
The preparation of diketone
By in embodiment 1 3) preparation method of item by 261mg (0.4mmol) 1- (3,4- bis--anisyl) -7- [(4- oxygen
Acetyl-Lys-OBzl) -3- methoxyphenyls] -1,6- heptadiene -3,5- diketone hydrolyzes to obtain 223mg (93%) title compound,
For yellow powder.ESI-MS(m/e)：569[M+H]+.1H NMR (300MHz, DMSO-d6)：δ/ppm=8.34-8.31 (m,
0.4H), 7.97 (bs, 0.7H), 7.61 (s, 1H), 7.56 (s, 1H), 7.39-7.28 (m, 4H), 7.03-6.82 (m, 4H),
6.13 (s, 1H), 4.64 (s, 2H), 4.25 (m, 1H), 3.87 (s, 3H), 3.83 (s, 3H), 3.81 (s, 3H), 2.76-2.71
(m, 2H), 1.74 (m, 2H), 1.54 (m, 2H), 1.33 (m, 2H).
Embodiment 6 prepares 1- (3- methoxyl group -4- hydroxyphenyls) -7- [4- oxygen acetyl-Lys (Pro-Ala-Lys) -3- methoxies
Base phenyl] -1,6- heptadiene -3,5- diketone (7a)
1) 1- (3- methoxyl group -4- hydroxyphenyls) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)]-OBzl-3-
Methoxyphenyl } -1,6- heptadiene -3,5- diketone preparation
By in embodiment 1 2) preparation method of item by 5.08g (10mmol) Boc-Pro-Ala-Lys (Boc) and 4.4g
(10mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- [(4- oxygen acetyl-Lys-Obzl) -3- methoxyphenyls] -1,6- heptan two
Bis- reactive ketone of alkene -3,5- obtains 7.02g (94%) title compound, is yellow powder.ESI-MS(m/e)：1141[M+H]+.1H
NMR (300MHz, DMSO-d6)：δ/ppm=7.63-7.53 (m, 3H), 7.35 (s, 5H), 7.25 (bs, 0.5H), 7.14-7.07
(m, 4H), 6.96-6.90 (m, 2H), 6.79 (m, 1H), 6.60-6.48 (m, 3H), 5.84 (s, 1H), 5.18 (s, 2H), 4.68-
4.57 (m, 4H), 4.36-4.18 (m, 3H), 3.95 (s, 3H), 3.92 (s, 3H), 3.50-3.48 (m, 2H), 3.17 (m, 2H),
3.08 (m, 2H), 2.19-1.25 (m, 42H).
2) 1- (3- methoxyl group -4- hydroxyphenyls) -7- { [4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)] -3- methoxies
Base phenyl } -1,6- heptadiene -3,5- diketone preparation
By in embodiment 1 3) preparation method of item by 456mg (0.4mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- { 4-
Oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone hydrolyzes
371mg (93%) title compound is yellow powder.ESI-MS(m/e)：1051[M+H]+。
3) 1- (3- methoxyl group -4- hydroxyphenyls) -7- [4- oxygen acetyl-Lys (Pro-Ala-Lys) -3- methoxyphenyls] -1,
The preparation of 6- heptadiene -3,5- diketone
By the preparation method in embodiment 4 2) by 371mg (0.36mmol) 1- (3- methoxyl group -4- hydroxyphenyls) -7- { 4- oxygen
Acetyl-Lys [Boc-Pro-Ala-Lys (Boc)] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone removing Boc obtain
315mg (98%) title compound is yellow powder.ESI-MS(m/e)：851[M+H]+.1H NMR (300MHz, DMSO-d6)：
δ/ppm=8.74-8.72 (m, 2H), 8.02-7.96 (m, 5H), 8.00-6.66 (m, 12H), 6.10 (s, 1H), 4.60 (s,
2H), 4.35 (m, 1H), 4.33-4.08 (m, 3H), 3.93 (s, 3H), 3.81 (s, 3H), 3.20 (m, 2H), 3.10-2.95 (m,
2H), 2.70 (m, 2H), 2.33-2.22 (m, 1H), 1.86 (m, 6H), 1.76-1.54 (m, 7H), 1.52-1.19 (m, 9H).
Embodiment 7 prepares 1- (3,4- bis--anisyl) -7- [4- oxygen acetyl-Lys (Pro-Ala-Lys) -3- methoxyl groups
Phenyl] -1,6- heptadiene -3,5- diketone (7b)
1) 1- (3,4- bis--anisyl) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)-OBzl] -3- first
Phenyl } -1,6- heptadiene -3,5- diketone preparation
By in embodiment 1 2) preparation method of item by 4.6g (10mmol) Boc-Pro-Ala-Lys (Boc) and 5.08g
(10mmol) 1- (3,4- bis--anisyl) -7- [(4- oxygen acetyl-Lys-OBzl) -3- methoxyphenyls] -1,6- heptadiene -
3,5- bis- reactive ketones obtain 7.24g (94%) title compound, are yellow powder.ESI-MS(m/e)：1155[M+H]+.1H NMR
(300MHz, CDCl3-d1)：δ/ppm=7.66-7.52 (m, 3H), 7.36 (m, 5H), 7.17-7.10 (m, 4H), 6.95-6.89
(m, 2H), 6.74 (m, 1H), 6.57-6.51 (m, 3H), 5.87 (s, 1H), 5.19 (m, 2H), 4.68-4.56 (m, 3H), 4.37-
4.16 (m, 3H), 3.96 (s, 3H), 3.95 (s, 3H), 3.93 (s, 3H), 3.49 (m, 2H), 3.17-3.07 (m, 4H), 2.23
(m, 2H), 2.08 (m, 2H), 1.92-1.77 (m, 5H), 1.48-1.35 (m, 30H).
2) 1- (3,4- bis--anisyl) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)] -3- methoxyl groups
Phenyl } -1,6- heptadiene -3,5- diketone preparation
By in embodiment 1 3) preparation method of item by 465mg (0.4mmol) 1- (3,4- bis--anisyl) -7- { 4- oxygen
Acetyl-Lys [Boc-Pro-Ala-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone hydrolyzes
401mg (93%) title compound is yellow powder.ESI-MS(m/e)：1065[M+H]+。
3) 1- (3,4- bis--anisyl) -7- [4- oxygen acetyl-Lys (Pro-Ala-Lys) -3- methoxyphenyls] -1,6-
The preparation of heptadiene -3,5- diketone
By in embodiment 4 2) preparation method of item by 213mg (0.2mmol) 1- (3,4- bis--anisyl) -7- { 4- oxygen
Acetyl-Lys [Boc-Pro-Ala-Lys (Boc)] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone removing Boc obtain
152mg (88%) title compound is yellow powder.ESI-MS(m/e)：865[M+H]+.1H NMR (300MHz, DMSO-d6)：
δ/ppm=8.74-8.72 (m, 2H), 8.02-7.96 (m, 5H), 8.00-6.66 (m, 12H), 6.10 (s, 1H), 4.60 (s,
2H), 4.35 (m, 1H), 4.33-4.08 (m, 3H), 3.93 (s, 3H), 3.81 (s, 3H), 3.20 (m, 2H), 3.10-2.95 (m,
2H), 2.70 (m, 2H), 2.33-2.22 (m, 1H), 1.86 (m, 6H), 1.76-1.54 (m, 7H), 1.52-1.19 (m, 9H).
The thrombus dissolving activity of the compound intravenous injection of 8 general formula 1 of embodiment
1) evaluation method
200-220g male SD rats 20% urethane solution (6mL/kg, i.p.) is anaesthetized.Anesthetized rat is faced upward
Clinostatism is fixed, and right common carotid artery is detached, and presss from both sides artery clamp in proximal part, proximal part and distal end respectively penetrate surgical thread, by telecentricity
The surgical thread at end is clamped in fur with haemostatic clamp, is intubated in distal end, is unclamped artery clamp, is released about 1mL arterial bloods and is mounted in 1mL
EP pipes in.Toward injection in vertical fixed glass tube (long 15mm, internal diameter 2.5mm, outer diameter 5.0mm, tube bottom are sealed with rubber plug)
0.1mL rat artery blood is rapidly inserted into the thrombus fixing bolt of a stainless steel material in pipe.The thrombus is fixed spiral and is used
The stainless steel wire of a diameter of 0.2mm is coiled into, the long 12mm of spiral part, contain 15 bung flanges, a diameter of 1.0mm of bung flange, support handle with
Spiral is connected, long 7.0mm, is in question mark type.After blood clotting 15min, the rubber plug of glass bottom of the tube is opened, thrombus is fixed with tweezers
The support handle of fixed spiral takes out the thrombus wrapped up by thrombus from glass tube and fixes spiral, is precisely weighed.
Bypass intubation is constituted by 3 sections, and stage casing is polyethylene rubber tube, long 60mm, internal diameter 3.5mm, and both ends are identical poly- second
One end of alkene pipe, long 100mm, internal diameter 1mm, outer diameter 2mm, the pipe pulls into spike tube (for being inserted into rat carotid artery or vein), outside
Diameter 1mm, polyethylene pipe (overstriking, the polyethylene glue for being inserted into stage casing of outer cover one section of long 7mm, outer diameter 3.5mm of the other end
In pipe).The equal silanization of inner wall of 3 sections of pipes.The thrombus of thrombus package is fixed spiral to be put into the polyethylene rubber tube of stage casing, sebific duct
Overstriking end of the both ends respectively with two polyethylene is nested.With syringe heparin-saline solution will be filled by spike tube end in pipe
(50IU/kg) is spare.
The left vena jugularis externa of rat is detached, proximal part and distal end respectively penetrate surgical thread, distal end are ligatured, exposed
An angle is carefully cut on left vena jugularis externa, and the spike tube of good bypass duct prepared above is inserted into left vena jugularis externa by angle and is opened
The proximal part of mouth, while far from the support handle of shunt valve stage casing (fixing spiral containing the thrombus being precisely weighed) interior thrombus fixation spiral.
The heparin-saline (50IU/kg) of correct amount is pushed by the spike tube of the other end with syringe, syringe is not withdrawn poly- at this time
Ethylene tube clamps the hose between syringe and polyethylene pipe with haemostatic clamp.In the proximal part artery pinching of right common carotid artery
Right common carotid artery is nearby carefully being cut an angle by blood from artery clamp.Syringe is extracted from the tip of polyethylene pipe, will be gathered
The proximal part of artery angle is inserted into the tip of ethylene tube.The both ends of bypass duct are all fixed with No. 4 sutures with arteriovenous.
With scalp acupuncture by physiological saline, the normal saline solution of urokinase or the normal saline solution of various concentration compound
By the stage casing (fixing spiral containing the thrombus being precisely weighed) of shunt valve, it is pierced into and is fixed at the nearly vein of spiral far from thrombus, beaten
Artery clamp is opened, so that blood flow is passed through bypass duct and flows to vein from artery, i.e. rat arteriovenous bypasses Thrombolysis Model for this, slowly will note
Liquid in emitter is injected into blood (about 6min), and physiological saline, urokinase or the compound of the present invention is made to be followed by blood
Ring, in the sequential action to thrombus by vein-heart-artery.From timing when starting injection, blood is taken out from bypass duct after 1h
Bolt fixes spiral, is precisely weighed.It calculates thrombus in every rat bypass duct and fixes front and back of poor quality of spiral administration, statistics is simultaneously
Evaluate the internal thrombolysis activity of compound.Thrombus loss of weight mean value and standard deviationIt indicates.
2) dosage
Physiological saline (blank control) dosage is 3mL/kg, and urokinase (positive control) dosage is equivalent to for 20000U/kg
The dosage of 1.68mg/kg, compounds of formula I 5a and 5b are 10nmol/kg, and the dosage of compounds of formula I 7a and 7b are
1.0nmol/kg。
3) evaluation results are shown in Table 1.The result shows that compounds of formula I 5a, 5b, 7a and 7b have outstanding thrombus dissolving to live
Property.
The thrombus dissolving activity a of 1 compound intravenous administration of table administration
A) n=10；B) with physiological saline group ratio, P ＜ 0.01.
Embodiment 9 is injected intravenously the thrombus dissolving activity of the compound 5a and 7a of various dose general formula 1
1) evaluation method is the same as 1) item in experimental example 1.
2) dosage
Choose the dosage effect dependence that tri- dosage of 100nmol/kg, 10nmol/kg, 1nmol/kg investigate 5a.Choosing
Tri- dosage of 10nmol/kg, 1nmol/kg, 0.1nmol/kg are taken to investigate the dosage effect dependence of 7a.
3) the dosage effect dependence evaluation result of 5a is shown in Table 2.The result shows that the thrombus dissolving activity of 5a is presented with dosage
Correlativity.
Table 2 is injected intravenously the dose-effect relationship of 5aa
A) n=10；B) with 10nmol/kg group ratios, p ＜ 0.05；C) with 1nmol/kg group ratios, p ＜ 0.01；And physiology salt d)
Water group ratio, p ＞ 0.05.
4) the dosage effect dependence evaluation result of 7a is shown in Table 3.The result shows that the thrombus dissolving activity of 7a is presented with dosage
Correlativity.
Table 3 is injected intravenously the dose-effect relationship of 7aa
A) n=10；B) with 1nmol/kg group ratios, p ＜ 0.05；C) with 0.1nmol/kg group ratios, p ＜ 0.01；And physiology d)
Brine group ratio, p ＞ 0.05.
The hemolysis in vitro thrombus activity of the compound of 10 general formula 1 of embodiment
1) evaluation method
Male SD rat, 250 ± 10g weigh, and intraperitoneal injection 20% urethane anesthesia, dorsal position is fixed.Isolate rat
Right common carotid artery, press from both sides artery clamp in proximal part, proximal part and distal end respectively penetrate surgical thread, ligature distal end, from dynamic
Right common carotid artery is nearby carefully cut an angle by arteries and veins folder, and the one end for being inserted into hexasilane pulls into the polyethylene pipe of spike tube, pine
Artery clamp is opened, release about 5mL arterial bloods every time and in the EP pipes of the hexasilane of 5mL, can about be put 3 times.Use hexasilane
The past each vertical fixed glass tube of the 5mL syringes of change (long 18mm, internal diameter 4mm, outer diameter 5.5mm, tube bottom are sealed with rubber plug)
Middle injection 0.1mL rat artery blood, is rapidly inserted into the thrombus fixing bolt of a stainless steel material in pipe.The thrombus is fixed
Spiral is coiled into the stainless steel wire of a diameter of 0.2mm, the long 20mm of spiral part, a diameter of 1.0mm of bung flange, holds in the palm handle and spiral phase
Even, long 7.0mm is in question mark type.After blood clotting 40min, the rubber plug of glass bottom of the tube is opened, thrombus is fixed with tweezers and fixes spiral shell
The support handle of rotation takes out the thrombus wrapped up by thrombus from glass tube and fixes spiral, and the cillin bottle for filling 8mL high purity waters is immersed in suspension
In, 1h is impregnated, removes and floats blood, taking-up is dipped in dry, is precisely weighed, and is recorded.It is certain that thrombus fixation coil suspension immersion is filled into 8mL again
In the cillin bottle of the compound solution of concentration, 37 DEG C of shaking tables are incubated 2h, and taking-up is dipped in dry, then is precisely weighed, is recorded.Thrombus fixes spiral shell
The front and back as thrombus loss of weight of poor quality of rotation, counts and evaluates the internal thrombolysis activity of compound.Thrombus loss of weight mean value and mark
It is accurate poorIt indicates.
2) medication and concentration
According to the corresponding blood concentration of internal thrombus dissolving dosage conversion.Blank control is physiological saline, and positive control is
UK (a concentration of 254 μ g/1ml), passes through a concentration of 10nM of a concentration of 100nM, 7a and 7b of the compound 5a and 5b of Formulas I.
3) evaluation result is shown in Table 4.The result shows that compounds of formula I shows outstanding thrombus dissolving activity in vitro.
The hemolysis in vitro thrombus activity of table 4 5a, 5b, 7a and 7ba
A) n=6；B) with physiological saline group ratio, P ＜ 0.01.
The antithrombotic acitivity of the compound intravenous injection of 11 general formula 1 of embodiment
1) evaluation method
Intubation is constituted by 3 sections, middle segment length 80mm, internal diameter 3.5mm, and both ends are identical polyethylene pipe, long 100mm, internal diameter
One end of 1mm, outer diameter 2mm, the pipe pull into spike tube (for being inserted into rat carotid artery or vein), the equal silanization of inner wall of 3 sections of pipes.
The silk thread of the long 60mm to weigh in advance is put into the polyethylene extra heavy pipe of stage casing, the both ends of extra heavy pipe respectively with two polyethylene tubules
End is not drawn and attenuated to be nested (silk thread is pushed down 0.5mm and fixed by wherein one section).With syringe heparin life will be filled by spike tube end in pipe
It is spare to manage saline solution (50IU/kg).
200-220g male SD rats 20% urethane solution (6mL/kg, i.p.) is anaesthetized.Anesthetized rat is faced upward
Clinostatism is fixed, and the left vena jugularis externa of rat is isolated, and proximal part and distal end respectively penetrate surgical thread, distal end are ligatured, sudden and violent
An angle is carefully cut on the left vena jugularis externa of dew, the non-crimping end spike tube of good bypass duct prepared above is inserted by angle
The proximal part of left vena jugularis externa opening, the heparin-saline (50IU/ of correct amount is pushed into syringe by the spike tube of the other end
Kg), syringe is pulled up, then suction is had into physiological saline, the normal saline solution of urokinase or the physiology salt of various concentration compound
The syringe of aqueous solution is inserted into the tip of polyethylene pipe, is pushed into liquid by the tip of polyethylene pipe, syringe is not withdrawn at this time
Polyethylene pipe detaches right common carotid artery, presss from both sides artery clamp in proximal part, proximal part and distal end respectively penetrate surgical thread, and ligation is remote
Right common carotid artery is nearby carefully being cut an angle by heart end from artery clamp.Syringe is extracted from the tip of polyethylene pipe, it will
The proximal part of artery angle is inserted into the tip of polyethylene pipe.The both ends of bypass duct are all fixed with No. 4 sutures with arteriovenous.
Artery clamp is opened, so that blood flow is passed through bypass duct and flows to vein from artery, i.e. rat arteriovenous bypasses anti-bolt model for this.From the beginning of
Timing when cycle is taken out the silk thread for hanging with thrombus after 15min from bypass duct, is precisely weighed, before and after silk thread it is of poor quality i.e.
For wet weight of thrombus, the internal anti-thrombus activity of compound is counted and evaluated.Wet weight of thrombus mean value and standard deviationTable
Show.
2) dosage
Physiological saline (blank control) dosage is 1mLkg-1, aspirin (positive control) dosage is 9mg/kg, general formula I
The dosage of compound 5a and 5b be 10nmol/kg, the dosage of 7aization 7b is 1.0nmol/kg.
3) evaluation result is shown in Table 5.The result shows that the administration of compounds of formula I intravenous administration can show outstanding antithrombotic
Activity.
The antithrombotic acitivity of 5 compound intravenous administration of table administration
A) n=10；B) with physiological saline group ratio, P ＜ 0.01.
The anti-inflammatory activity of the compound oral medication of 12 general formula 1 of embodiment
1) evaluation method
18-22g ICR male mices are randomly divided into blank control group, positive medication group and administration group, and mouse uses preceding quiet
Breath 1 day, operation room keep 22 DEG C of indoor temperature, every group of mouse 10.Experiment distinguishes gastric infusion when starting, and single administration 30 divides
Dimethylbenzene (0.03mL) is applied toward the left ear gabarit of small white mouse after clock, puts to death small white mouse cervical dislocation after 2 hours.By mouse
A left side, auris dextra are cut, and with the card punch of diameter 7mm in the same position of two ears, are taken round auricle, are weighed respectively, find out two circle ears
The weight difference of piece is as swelling.Swelling=left ear original sheet weight-auris dextra original sheet weight.
2) dosage
Physiological saline (blank control) dosage is 10mL/kg, and aspirin (positive control) dosage is 200mg/kg, general formula
The dosage of the compound 5a and 5b of I are that the dosage of 10nmol/kg, 7a and 7b are 1.0nmol/kg.
3) evaluation result is shown in Table 6.The result shows that compounds of formula I oral administration shows outstanding anti-inflammatory activity.
The oral anti-inflammatory activity of 6 compound of tablea
A) n=10, aspirin dose 200mg/kg；B) with physiological saline group ratio, P ＜ 0.01.
The activity of the compound scavenging hydroxyl of 13 general formula 1 of experimental example
1) evaluation method
According to the corresponding blood concentration of internal thrombus dissolving dosage conversion.Compounds of formula I 5a and 5b add distilled water
It is made into the solution of a concentration of 100nM, compound 7a and 7b add distilled water to be made into the solution of a concentration of 10nM, on EPR spectrometer
It measures.
According to standard method by FeSO4·7H2O adds distilled water to be made into the solution A of a concentration of 10nM, by 30%H2O2Add steaming
Distilled water is made into a concentration of 1%H2O2Solution B, DMPO added distilled water be made into the solution C of a concentration of 1.1316mg/100mL.
Blank control group adds 2.5 μ L solution As, 5 μ L solution Cs, 5 μ L distilled water and 2.5 μ L solution Bs successively, after being vigorously mixed
Quartz capillary is sucked, hydroxy radical signal is measured after 1min and records spectrogram.
The compound group of general formula 1 adds the change of the general formula 1 of 2.5 μ L solution As, 5 μ L solution Cs, the compound of 5 μ L general formulas 1 successively
Object and 2.5 μ L solution Bs are closed, quartz capillary is sucked after being vigorously mixed, hydroxy radical signal is measured after 1min and records spectrogram.
1 sample is surveyed 5 times under 1 concentration, is recorded each spectrogram peak height, calculates Scavenging action to hydroxyl free radical.It calculates public
Formula is：Clearance rate=(the compound group peak height of blank control group peak height-general formula 1)/blank control group peak height.
2) evaluation result is shown in Table 7.The result shows that compounds of formula I is outstanding hydroxy radical scavenger.
The percentage of 7 compound scavenging hydroxyl of table
The nanostructure of the compound of 14 general formula 1 of embodiment
1) when observing 25 DEG C and 37 DEG C on laser diffraction particle size instrument, the aqueous solution of compounds of formula I and pH are 7.4
Phosphate buffer solution grain size.The concentration of compound converts according to internal thrombus dissolving dosage, and 5a and 5b's is a concentration of
100nM the results are shown in Table 8.A concentration of 10nM. of 7a and 7b, the results are shown in Table 9.Statistics indicate that compounds of formula I is in water and phosphorus
Hydrochlorate buffer solution has suitable nanometer spherical structure.
The sphere diameter for the nanosphere that 8 compound of table is formed in aqueous solution
| Compound | Nanometer sphere diameter (nm, 25 DEG C) | Nanometer sphere diameter (nm, 37 DEG C) |
| 5a | 236.5 | 137.8 |
| 5b | 217.6 | 148.0 |
| 7a | 206.4 | 121.7 |
| 7b | 317.2 | 122.6 |
The sphere diameter for the nanosphere that 9 compound of table is formed in the phosphate buffer solution that pH is 7.4
| Compound | Nanometer sphere diameter (nm, 25 DEG C) | Nanometer sphere diameter (nm, 37 DEG C) |
| 5a | 208.8 | 176.4 |
| 5b | 228.8 | 178.7 |
| 7a | 241.8 | 190.0 |
| 7b | 311.3 | 261.4 |
2) it is 10 to 4 compound concentrations for determining general formula I under transmission electron microscope-7The transmission electricity of the aqueous solution of mg/mL
Mirror photo.7b also measured were 10 as representing-6、10-7、10-8The transmission electron microscope photo of the aqueous solution of tri- concentration of mg/mL.Knot
Fruit shows that nanosphere, nanometer rods, nano-rings, nanotube etc. can be observed no under transmission electron microscope in 4 compounds of general formula I
Same nanostructure.Different nanostructures can be observed in 7b under the conditions of various concentration and pH.The transmission of 5a, 5b, 7a and 7b
Electromicroscopic photograph is shown in Fig. 2, Fig. 3, Fig. 4 and Fig. 5 respectively.
Claims (8)
1. the pseudo- peptide of following formula
2. the method for preparing the pseudo- peptide of claim 1, this method are made of following steps：
(1) in the presence of dicyclohexylcarbodiimide (DCC) and n-hydroxysuccinimide (HOSu) Boc-Pro in anhydrous THF
In with HOSu condensation be Boc-Pro-OSu, in NaHCO3In the presence of Boc-Pro-OSu reacted with L-Ala generate Boc-Pro-
Ala；
(2) Boc-Pro-Ala is Boc-Pro- with Lys (Boc)-OBzl condensations in anhydrous THF in the presence of DCC and HOBt
Ala-Lys(Boc)-OBzl；
(3) Boc-Pro-Ala-Lys (Boc)-OBzl is hydrolyzed to Boc-Pro-Ala-Lys in methyl alcohol in the presence of NaOH
(Boc)；
(4) in anhydrous K2CO3In the presence of in anhydrous THF vanillic aldehyde reacted with bromoacetate generate 3- methoxyl group -4- oxygen acetyl
Ethyl ester benzaldehyde；
(5) in B2O3、(nBuo)3In the presence of B and nBu-NH Veratraldehyde in anhydrous ethyl acetate with levulinic
Ketone condensation is 6- (3,4- bis--anisyl) -5,6- hexene -2,4- diketone；
(6) in B2O3、(nBuo)3In the presence of B and nBu-NH, the 6- (4- methyl -3- methoxyphenyls)-in anhydrous ethyl acetate
5,6- hexene -2,4- diketone are 1- (4- methyl -3- methoxybenzenes with the condensation of 3- methoxyl group -4- oxygen acetyl triethyl benzaldehydes
Base) -7- (4- oxygen acetyl triethyl base -3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(7) in the presence of NaOH, in acetone by 1- (4- methyl -3- methoxyphenyls) -7- (4- oxygen acetyl triethyl base -3- first
Phenyl) -1,6- heptadiene -3,5- diketone is hydrolyzed to 1- (4- methyl -3- methoxyphenyls) -7- (4- fluoroacetic acid base -3- first
Phenyl) -1,6- heptadiene -3,5- diketone；
(8) in the presence of DCC and HOBt in anhydrous THF 1- (4- methyl -3- methoxyphenyls) -7- (4- fluoroacetic acid base -3- first
Phenyl) -1,6- heptadiene -3,5- diketone and Lys (Boc)-OBzl condensation is 1- (4- methyl -3- methoxyphenyls) -7-
(4- oxygen acetyl-Lys (Boc)-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(9) 1- (4- methyl -3- methoxyphenyls) -7- (the 4- oxygen acetyl-Lys (Boc)-in hydrogen chloride-ethyl acetate solution
OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone slough Boc generate 1- (4- methyl -3- methoxyphenyls) -7-
(4- oxygen acetyl-Lys-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone；
(10) in the presence of DCC and HOBt in anhydrous THF Boc-Pro-Ala-Lys (Boc) and 1- (4- methyl -3- methoxyl groups
Phenyl) -7- (4- oxygen acetyl-Lys-OBzl-3- methoxyphenyls) -1,6- heptadiene -3,5- diketone condensation be 1- (4- methyl -
3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,6- heptan
Diene -3,5- diketone；
(11) in methyl alcohol by 1- (4- methyl -3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys [Boc- in the presence of NaOH
Pro-Ala-Lys (Boc)-OBzl] -3- methoxyphenyls } -1,6- heptadiene -3,5- diketone is hydrolyzed to 1- (4- methyl -3- first
Phenyl) -7- { 4- oxygen acetyl-Lys [Boc-Pro-Ala-Lys (Boc)] -3- methoxyphenyls } -1,6- heptadiene -3,5-
Diketone；
(12) by 1- (4- methyl -3- methoxyphenyls) -7- { 4- oxygen acetyl-Lys [Boc- in hydrogen chloride-ethyl acetate solution
Pro-Ala-Lys (Boc)] -3- methoxyphenyls -1,6- heptadiene -3,5- diketone slough Boc generate 1- (4- methyl -3- first
Phenyl) -7- [4- oxygen acetyl-Lys (Pro-Ala-Lys) -3- methoxyphenyls] -1,6- heptadiene -3,5- diketone.
3. the pseudo- peptide of claim 1 prepares the application of anti-inflammatory drug.
4. the pseudo- peptide of claim 1 prepares the application of OH radicals scavenging drugs.
5. the pseudo- peptide of claim 1 prepares the application of thrombolytic agent.
6. the pseudo- peptide of claim 1 prepares the application of thrombus dissolving and antithrombotic reagent.
7. the pseudo- peptide of claim 1 prepares the application of thrombus dissolving, antithrombotic and anti-inflammatory drug.
8. the pseudo- peptide of claim 1 prepares thrombus dissolving, antithrombotic, anti-inflammatory and OH radicals scavenging drugs application.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410562048.1A CN104610424B (en) | 2011-12-13 | 2011-12-13 | Lys (Pro-Ala-Lys) curcumin derivate, its synthesis and the application in medicine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410562048.1A CN104610424B (en) | 2011-12-13 | 2011-12-13 | Lys (Pro-Ala-Lys) curcumin derivate, its synthesis and the application in medicine |
| CN201110415171.7A CN103159826B (en) | 2011-12-13 | 2011-12-13 | Lys and Lys (Pro-Ala-Lys)-modified curcumin derivatives, and synthesis and medical application thereof |
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| 姜黄素衍生物与类似物的构效关系研究进展;韦星船等;《化学研究与应用》;20100531;第22卷(第5期);527-538 * |
| 寡肽药物先导结构的发现与优化;赵明等;《北京大学学报（医学版）》;20021031;第34卷(第5期);506-512 * |
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