CN101190940B - Polypeptide with target antithrombotic activity and its preparation method and application - Google Patents

Polypeptide with target antithrombotic activity and its preparation method and application Download PDF

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CN101190940B
CN101190940B CN2006101442349A CN200610144234A CN101190940B CN 101190940 B CN101190940 B CN 101190940B CN 2006101442349 A CN2006101442349 A CN 2006101442349A CN 200610144234 A CN200610144234 A CN 200610144234A CN 101190940 B CN101190940 B CN 101190940B
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gly
obzl
boc
arg
ser
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CN101190940A (en
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彭师奇
赵明
王超
赵婧华
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Capital Medical University
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Capital Medical University
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Abstract

The invention discloses polypeptides of the same class, which have targeted antithrombotic activity and the preparation method of the polypeptides thereof and the application thereof as antithrombotic agents, pertaining to the field of biological medicines. By conjugated embellishing on RGD peptide and YIGS peptide, the invention obtains the polypeptides which have targeted antithrombotic activity. The animal model test verifies that the polypeptides of the invention have comparatively strong antithrombotic activity.

Description

Polypeptide, its preparation method and application with target antithrombotic activity
Technical field
The present invention relates to polypeptide, relate in particular to a series of polypeptide that have target antithrombotic activity through manually modified and synthetic.The preparation method and they that the invention still further relates to these polypeptide belong to biomedicine field as the application of pharmaceutical preparations having antithrombotic activity.
Background technology
Cell adhesion plays a crucial role in the evolution of cell adhesion property disease (metastasis of cancer, thrombosis, chemistry cause inflammation and osteoporosis).Modulability gp for example laminine, Fibrinogen, RGD peptide, YIGSR and YIGSK etc. can be participated in the cell adhesion process.They and integrin receptor have very strong binding ability.For example in the platelet aggregation process, RGDS and RGDF have combined crucial effects with activatory platelet surface acceptor GP IIb/IIIa.The RGD peptide combines with the specificity of activatory platelet surface acceptor GP IIb/IIIa, has given the compound that contains the RGD sequence a kind of critical nature, and the compound that promptly contains the RGD sequence can be to the enrichment of the position of platelet activation.Under such prerequisite, put together the RGD peptide with anti-cell adherent YIGSR and YIGSK, just can obtain the target anti-cell and stick polypeptide.
Summary of the invention
One of the object of the invention is that RGD peptide and YIGS peptide are puted together the formula modification, prepares the polypeptide with target antithrombotic activity.
One of the object of the invention is realized through following technical scheme:
Polypeptide with target antithrombotic activity is selected from the polypeptide of following any one aminoacid sequence:
YIGSRRGDS、YIGSRRGDV、YIGSRRGDF、YIGSRYIGSK、YIGSRYIGSR、YIGSKRGDS、YIGSKRGDF、YIGSKRGDV、YIGSKYIGSK、YIGSKYIGSR、RGDSRGDS、RGDVRGDV、RGDFRGDF、RGDSYIGSR、RGDSYIGSK、RGDVYIGSR、RGDVYIGSK、RGDFYIGSR、RGDFYIGSK。
The present invention is directed to the advantage and the clinical demand of target antithrombotic medicine to the target antithrombotic medicine; According to RGD peptide, YIGSR and YIGSK etc. can participate in the cell adhesion process, according to they and integrin receptor have very strong binding ability, according in the platelet aggregation process RGD peptide combined with activatory platelet surface acceptor GP IIb/IIIa crucial effects, according to the RGD peptide combine with the specificity of activatory platelet surface acceptor GP IIb/IIIa the acquisition target property; RGD peptide and YIGS peptide are puted together the formula modification, prepare above-mentioned polypeptide with target antithrombotic activity.
Two of the object of the invention provides a kind of method for preparing the polypeptide of above-mentioned antithrombotic acitivity.
Two of the object of the invention is to realize through following technical scheme:
A kind of method for preparing the polypeptide of above-mentioned antithrombotic acitivity comprises:
1. employing liquid-phase synthesis process is through progressively connecing the protection midbody of the synthetic respectively RGDS of peptide, RGDF, RGDV, YIGSR and YIGSK;
2. slough the N end protection base of RGDS, RGDF, RGDV, YIGSR and YIGSK protection midbody respectively;
3. slough the C end protection base of RGDS, RGDF, RGDV, YIGSR and YIGSK protection midbody respectively;
4. (a) puts together formula to the RGDS of the RGDS of N end dissociative protection midbody and C end dissociative, YIGSR and YIGSK protection midbody respectively and modifies, slough respectively again to put together the formula modification and obtain RGDSRGDS, and RGDSYIGSR,
RGDSYIGSK three peptide species; (b) put together formula to the RGDF of the RGDF of N end dissociative protection midbody and C end dissociative, YIGSR and YIGSK protection midbody respectively and modify, slough respectively again and put together the formula modification and obtain RGDFRGDF, RGDFYIGSR, RGDFYIGSK three peptide species; (c) put together formula to the RGDV of the RGDV of N end dissociative protection midbody and C end dissociative, YIGSR and YIGSK protection midbody respectively and modify, slough respectively again and put together the formula modification and obtain RGDVRGDV, RGDVYIGSR, RGDVYIGSK three peptide species; (d) put together formula to the RGDS of the YIGSR of N end dissociative protection midbody and C end dissociative, RGDF, RGDV, YIGSR and YIGSK protection midbody respectively and modify, slough respectively again and put together the formula modification and obtain YIGSRRGDS, YIGSRRGDV, YIGSRRGDF, YIGSRYIGSK, YIGSRYIGSR five peptide species; (e) put together formula to the RGDS of the YIGSK of N end dissociative protection midbody and C end dissociative, RGDF, RGDV, YIGSR and YIGSK protection midbody respectively and modify, slough again and put together the formula modification and obtain YIGSKRGDS, YIGSKRGDF, YIGSKRGDV, YIGSKYIGSK, YIGSKYIGSR five peptide species.
Wherein, described N end protection base is the blocking group that the N end of polypeptide is used always when protecting, and for example can be tertbutyloxycarbonyl (Boc); Described C end protection base is the blocking group that the C end of polypeptide is used always when protecting, and for example can be benzyloxy (OBzl), methoxyl group (OMe) etc.
Above-described liquid phase synthetic intermediate and the method for protecting are the conventional and technique known of this area.
Animal model test proves, puts together the formula modified polypeptide and has stronger antithrombotic acitivity for 19 kinds of the present invention.
In order further to set forth the present invention, provide a series of embodiment below.These embodiment are illustrative fully, and they only are used for the present invention is specifically described, and are not to be understood that to be limitation of the present invention.
The explanation of the shortenings that is occurred among the present invention:
The THF THF
DCC dicyclohexyl imide
The DCU NSC 30023
The OBzl benzyloxy
The Boc tertbutyloxycarbonyl
The OMe methoxyl group
HOBt N-hydroxybenzotriazole
The NMM N-methylmorpholine
Embodiment
The preparation of embodiment 1YIGSR
1) Boc-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.590g (2.0mmol) Boc-Ser (Bzl)-OH is dissolved among the anhydrous THF of 6ml, and 0 ℃ adds 0.270g (2.0mmol) N-hydroxybenzotriazole (HOBt) and 0.45g (2.2mmol) dicyclohexyl carbonyl diimine (DCC) down.Stir and add 0.539g (2.0mmol) HCl-NH after 10 minutes 2-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with N-methylmorpholine (NMM), and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the NSC 30023 (DCU) that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1), obtain 0.969g (95%) target compound.ESI-MS(m/z)511[M+H] +。
2) HClNH 2-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
With 0.969g (1.9mmol) Boc-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in 15ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)411[M+H] +。
3) preparation of Boc-Ile-Gly-OMe
With 0.498g (2.0mmol) Boc-Ile-OHH 2O is dissolved among the anhydrous THF of 5mL, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir and add 0.251g (2.0mmol) HClNH after 10 minutes 2The anhydrous THF solution of-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 60: 1).Obtain 0.567g (94%) target compound.ESI-MS(m/z)303[M+H] +。
4) preparation of HClIle-GlyOMe
0.567g (1.87mmol) Boc-Ile-Gly-OMe is dissolved in 15ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)203[M+H] +。
5) preparation of Boc-Tyr-Ile-Gly-OMe
0.525g (1.87mmol) Boc-Tyr-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.377g (1.87mmol) HClH-Ile-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 60: 1).Obtain 0.808g (93%) target compound.ESI-MS(m/z)466[M+H] +。
6) preparation of Boc-Tyr-Ile-Gly-OH
0.808g (1.87mmol) Boc-Tyr-Ile-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.834g (99%) target compound.ESI-MS(m/z)452[M+H] +。
7) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.857g (1.9mmol) Boc-Tyr-Ile-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.257g (1.9mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add 0.848g (1.9mmol) HClH-Ser-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.44g (90%) target compound.ESI-MS(m/z)844[M+H] +。
8) HClNH 2-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
With 1.44g (1.70mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in 20ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)743[M+H] +。
9) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OH
With 1.688g (2.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO 4Regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation, and residuary water solution continues to use KHSO 4Regulate pH value to 2, with ethyl acetate extraction three times, ethyl acetate layer merges, give a baby a bath on the third day after its birth time with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4, filtration, filtrate decompression is concentrated removes ETHYLE ACETATE, obtains 1.59g (96%) target compound.ESI-MS(m/z)829[M-H] -。
10) preparation of YIGSR
With 250mgBoc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 160mg (91%) target compound.ESI-MS(m/z)594[M+H] +。
The preparation of embodiment 2YIGSK
1) preparation of Boc-Ser (Bzl)-Lys (Z)-OBzl
0.590g (2.0mmol) Boc-Ser (Bzl)-OH is dissolved among the anhydrous THF of 6ml, and 0 ℃ adds 0.270g (2.0mmol) N-hydroxybenzotriazole (HOBt) and 0.45g (2.2mmol) dicyclohexyl carbonyl diimine (DCC) down.Stir the anhydrous THF solution that adds 1.084g (2.0mmol) TosOHH-Lys (Z)-OBzl after 10 minutes.Reaction mixture is regulated pH 8-9 with N-methylmorpholine (NMM), and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the NSC 30023 (DCU) that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1), obtain 1.216g (94%) target compound.ESI-MS(m/z)648[M+H] +。
2) HClNH 2The preparation of-Ser (Bzl)-Lys (Z)-OBzl
0.648g (1.0mmol) Boc-Ser (Bzl)-Lys (Z)-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)548[M+H] +。
3) preparation of Boc-Ile-Gly-OMe
With 0.498g (2.0mmol) Boc-Ile-OHH 2O is dissolved among the anhydrous THF of 5mL, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir the anhydrous THF solution that adds 0.251g (2.0mmol) HClH-Gly-OMe after 10 minutes.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 60: 1).Obtain 0.567g (94%) target compound.ESI-MS(m/z)303[M+H] +。
4) preparation of HClH-Ile-Gly-OMe
0.567g (1.87mmol) Boc-Ile-Gly-OMe is dissolved in 15ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)203[M+H] +。
5) preparation of Boc-Tyr-Ile-Gly-OMe
0.525g (1.87mmol) Boc-Tyr-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.377g (1.87mmol) HCl-H-Ile-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 60: 1).Obtain 0.808g (93%) target compound.ESI-MS(m/z)466[M+H] +。
6) preparation of Boc-Tyr-Ile-Gly-OH
0.808g (1.87mmol) Boc-Tyr-Ile-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.834g (99%) target compound.ESI-MS(m/z)450[M-H] -。
7) preparation of Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
0.857g (1.9mmol) Boc-Tyr-Ile-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.257g (1.9mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 1.039g (1.9mmol) HClH-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.71g (92%) target compound.ESI-MS(m/z)981[M+H] +。
8) HClNH 2The preparation of-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
0.98g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in 20ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears; ETHYLE ACETATE is taken out in decompression, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)881[M+H] +。
9) preparation of Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OH
0.784g (0.8mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 1.59g (96%) target compound.ESI-MS(m/z)889[M-H] -。
10) preparation of YIGSK
200mg (0.204mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains product 105mg (90%) target compound.ESI-MS(m/z)567[M+H] +。
The preparation of embodiment 3RGDF
1) preparation of Boc-Asp (OBzl)-Phe-OBzl
0.646g (2.0mmol) Boc-Asp (OBzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.854g (2.0mmol) Tos-OHH-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.07g (95%) target compound.ESI-MS(m/z)561[M+H] +。
2) preparation of HClH-Asp (OBzl)-Phe-OBzl
0.560g (1.0mmol) Boc-Asp (OBzl)-Phe-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)461[M+H] +。
3) Boc-Arg (NO 2The preparation of)-Gly-OMe
With 0.638g (2.0mmol) Boc-Arg (NO 2)-OH is dissolved in the 5ml dry DMF, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous DMF solution of 0.251g (2.0mmol) HClH-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.760g (97%) target compound.ESI-MS(m/z)391[M+H] +。
4) Boc-Arg (NO 2The preparation of)-Gly-OH
With 0.760g (1.9mmol) Boc-Arg (NO 2)-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.714g (99%) target compound.ESI-MS(m/z)375[M-H] -。
5) Boc-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Phe-OBzl
With 0.714g (1.89mmol) Boc-Arg (NO 2)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.255g (1.89mmol) HOBt and 0.428g (2.08mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.871g (1.89mmol) HClH-Asp (OBzl)-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.47g (95%) target compound.ESI-MS(m/z)820[M+H] +。
5) HClH-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Phe-OBzl
With 0.720g (1.0mmol) Boc-Arg (NO 2)-Gly-Asp (OBzl)-Phe-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)720[M+H] +。
6) preparation of Boc-Asp (OMe)-Phe-OBzl
0.494g (2.0mmol) Boc-Asp (OMe)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.854g (2.0mmol) Tos-OHH-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.960g (99%) target compound.ESI-MS(m/z)485[M+H] +。
7) preparation of HClH-Asp (OMe)-Phe-OBzl
0.960g (1.98mmol) Boc-Asp (OMe)-Phe-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)385[M+H] +。
8) preparation of Boc-Arg (Tos)-Gly-OMe
0.856g (2.0mmol) Boc-Arg (Tos)-OH is dissolved in the 5ml dry DMF, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous DMF solution of 0.251g (2.0mmol) HClH-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.960g (96%) target compound.ESI-MS(m/z)500[M+H] +。
9) preparation of Boc-Arg (Tos)-Gly-OH
0.960g (1.9mmol) Boc-Arg (Tos)-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation, and residuary water solution continues to use KHSO 4Regulate pH value to 2, with ethyl acetate extraction three times, ethyl acetate layer merges, with saturated NaCl come together give a baby a bath on the third day after its birth inferior, anhydrous Na SO 4Dry 3 hours, to filter, filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.903g (98%) target compound.ESI-MS(m/z)484[M-H] -。
11) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-OBzl
0.903g (1.86mmol) Boc-Arg (Tos)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.252g (1.86mmol) HOBt and 0.428g (2.08mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.784g (1.86mmol) HClH-Asp (OMe)-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.47g (93%) target compound.ESI-MS(m/z)852[M+H] +。
12) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-OH
0.852g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-OBzl is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO 4Regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.731g (96%) target compound.ESI-MS(m/z)760[M-H] -。
13) preparation of RGDF
200mg (0.235mmol) Boc-Arg (TOS)-Gly-Asp (OMe)-Phe-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 106mg (92%) target compound.ESI-MS(m/z)494[M+H] +。
The preparation of embodiment 4RGDV
1) preparation of Boc-Asp (OBzl)-Val-OBzl
0.646g (2.0mmol) Boc-Asp (OBzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.758g (2.0mmol) Tos-OHH-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.942g (95%) target compound.ESI-MS(m/z)512[M+H] +。
2) HClNH 2The preparation of-Asp (OBzl)-Phe-OBzl
0.512g (1.0mmol) Boc-Asp (OBzl)-Phe-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)412[M+H] +。
3) Boc-Arg (NO 2The preparation of)-Gly-OMe
With 0.638g (2.0mmol) Boc-Arg (NO 2)-OH is dissolved in the 5ml dry DMF, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous DMF solution of 0.251g (2.0mmol) HClH-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.760g (97%) target compound.ESI-MS(m/z)391[M+H] +。
4) Boc-Arg (NO 2The preparation of)-Gly-OH
With 0.760g (1.9mmol) Boc-Arg (NO 2)-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.714g (99%) target compound.ESI-MS(m/z)375[M-H] -。
5) Boc-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Val-OBzl
With 0.714g (1.89mmol) Boc-Arg (NO 2)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.255g (1.89mmol) HOBt and 0.428g (2.08mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.848g (1.89mmol) HClH-Asp (OBzl)-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.35g (93%) target compound.ESI-MS(m/z)771[M+H] +。
6) HClH-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Val-OBzl
With 0.770g (1.0mmol) Boc-Arg (NO 2)-Gly-Asp (OBzl)-Val-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)671[M+H] +。
7) preparation of Boc-Asp (OMe)-Val-OBzl
0.494g (2.0mmol) Boc-Asp (OMe)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.758g (2.0mmol) Tos-OHH-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.819g (94%) target compound.ESI-MS(m/z)437[M+H] +。
8) preparation of HClH-Asp (OMe)-Val-OBzl
0.819g (1.98mmol) Boc-Asp (OMe)-Val-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)337[M+H] +。
9) preparation of Boc-Arg (Tos)-Gly-OMe
0.856g (2.0mmol) Boc-Arg (Tos)-OH is dissolved in the 5ml dry DMF, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous DMF solution of 0.251g (2.0mmol) HClH-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.960g (96%) target compound.ESI-MS(m/z)500[M+H] +。
10) preparation of Boc-Arg (Tos)-Gly-OH
0.960g (1.9mmol) Boc-Arg (Tos)-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.903g (98%) target compound.ESI-MS(m/z)484[M-H] -。
11) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OBzl
0.903g (1.86mmol) Boc-Arg (Tos)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.252g (1.86mmol) HOBt and 0.428g (2.08mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.693g (1.86mmol) HClH-Asp (OMe)-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.39g (93%) target compound.ESI-MS(m/z)804[M+H] +。
12) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OH
0.804g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OBzl is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation, and residuary water solution continues to use KHSO 4Regulate pH value to 2, with ethyl acetate extraction three times, ethyl acetate layer merges, give a baby a bath on the third day after its birth time with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4, filtration, filtrate decompression is concentrated removes ETHYLE ACETATE, obtains 0.684g (96%) target compound.ESI-MS(m/z)712[M-H] -。
13) preparation of RGDV
200mg (0.249mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 99mg (91%) target compound.ESI-MS(m/z)446[M+H] +。
The preparation of embodiment 5RGDS
1) preparation of Boc-Asp (OBzl)-Ser (Bzl)-OBzl
0.646g (2.0mmol) Boc-Asp (OBzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.914g (2.0mmol) Tos-OHH-Ser (Bzl)-OBzl.Reaction mixture is regulated pH8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.08g (92%) target compound.ESI-MS(m/z)591[M+H] +。
2) preparation of HClH-Asp (OBzl)-Ser (Bzl)-OBzl
0.590g (1.0mmol) Boc-Asp (OBzl)-Ser (Bzl)-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)491[M+H] +。
3) Boc-Arg (NO 2The preparation of)-Gly-OMe
With 0.638g (2.0mmol) Boc-Arg (NO 2)-OH is dissolved in the 5ml dry DMF, and 0C adds 0.270g (2.0mmol) HOBt and 0.45g (2.18mmol) DCC down.Stir after 10 minutes, add 0.251g (2.0mmol) HClNH 2The anhydrous DMF solution of-Gly-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.760g (97%) target compound.ESI-MS(m/z)391[M+H] +。
4) Boc-Arg (NO 2The preparation of)-Gly-OH
With 0.760g (1.9mmol) Boc-Arg (NO 2)-Gly-OMe is dissolved in the 10ml methyl alcohol, and 0 ℃ of NaOH aqueous solution that adds 1.5mol/l is down regulated pH value to 11, stirring reaction 0.5hr, and the TLC detection reaction finishes.Add saturated KHSO4 and regulate pH value to 6～7, the salt that filtering is separated out, whole methyl alcohol are removed in the filtrate decompression distillation; Residuary water solution continues to regulate pH value to 2 with KHSO4, with ethyl acetate extraction three times, and the ethyl acetate layer merging; It is inferior to give a baby a bath on the third day after its birth with saturated NaCl collection, and dry 3 hours of anhydrous Na SO4 filters; Filtrate decompression concentrates and removes ETHYLE ACETATE, obtains 0.714g (99%) target compound.ESI-MS(m/z)375[M-H] -。
5) Boc-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl
With 0.714g (1.89mmol) Boc-Arg (NO 2)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.255g (1.89mmol) HOBt and 0.428g (2.08mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 1.18g (1.89mmol) HClH-Asp (OBzl)-Ser (Bzl)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.48g (91%) target compound.ESI-MS(m/z)865[M+H] +。
6) HClH-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl
With 0.865g (1.0mmol) Boc-Arg (NO 2)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)765[M+H] +。
7) preparation of RGDS
With 200mg (0.231mmol) Boc-Arg (NO 2)-Gly-Asp (Bzl)-Ser (Bzl)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 94mg (90%) target compound.ESI-MS(m/z)450[M+H] +。
The preparation of embodiment 6RGDFYIGSR
1) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.762g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add 0.780g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.32g (89%) target compound.ESI-MS(m/z)1487[M+H] +。 1H?NMR(BHSC-300，CDCl 3-d 3)δ＝7.75(d，J＝5.0Hz，9H)，7.87(d，J＝7.0Hz，2H)，7.34(d，J＝7.0Hz，2H)，7.19(m，10H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，2H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，3H)，4.63(s，2H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.67(s，6H)，3.05(d，J＝6.0Hz，4H)，2.7(d，J＝7.0Hz，6H)，2.5(m，1H)，2.35(s，3H)，2.0(s，4H)，1.79(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，8H)。Ultimate analysis C 68H 94N 16O 20S theoretical value C 54.90, H, 6.37, N 15.06. measured value C 54.75, H 6.51, and N 15.22.
2) preparation of RGDFYIGSR
200mg (0.134mmol) Boc-Arg (TOS)-Gly-Asp (OMe)-Phe-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 4mL trifluoracetic acid, adds 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide, and stirring reaction is 1 hour under 0 ℃ of condition of ice bath; Decompression was bled 5 minutes; Disposable adding 100mL ether, the product pressed powder of separating out use Sephadex G 10 to purify, and obtain product 129mg (90%). 1H NMR (BHSC-300, CDCl 3-d 3) δ=11.0 (s, 2H), 7.80 (d, J=5.0Hz, 8H), 7.21 (d, J=7.0Hz, 2H), 7.12 (m, 2H), 7.08 (t; J=7.0Hz, 1H), 6.95 (d, J=7.0Hz, 2H), 6.68 (d, J=7.0Hz, 2H), 5.7 (s, 2H), 5.0 (s; 1H), 4.92 (q, J=6.0Hz, 2H), 4.86 (q, J=6.0Hz, 1H), 4.62 (s, 1H), 4.50 (t, J=6.0Hz; 2H), 4.09 (d, J=6.0Hz, 4H), 4.03 (d, J=7.0Hz, 2H), 3.56 (m, 1H), 3.05 (d; J=6.0Hz, 4H), 2.73 (t, J=6.0Hz, 2H), 2.65 (q, J=6.0Hz, 4H), 2.5 (m, 1H); 2.0 (s, 9H), 1.79 (q, J=7.0Hz, 4H), 1.55 (m, 4H), 0.8-1.3 (m, J=8.0Hz, 8H).ESI-MS(m/z)1071[M+H] +。Ultimate analysis C 68H 94N 16O 20S theoretical value C 52.75, H, 6.69, N 19.63. measured value C 52.62, H 6.60, and N 19.50.
The preparation of embodiment 7RGDVYIGSR
1) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.713g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add 0.780g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.29g (89%) target compound. 1H?NMR(BHSC-300，CDCl 3-d 3)δ＝7.75(d，J＝5.0Hz，9H)，7.87(d，J＝7.0Hz，2H)，7.34(d，J＝7.0Hz，2H)，7.19(m，5H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，2H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，2H)，4.63(s，2H)，4.52(t，J＝6.0Hz，4H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.67(s，6H)，3.05(d，J＝6Hz，2H)，2.7(d，J＝7.0Hz，6H)，2.5(m，1H)，2.35(s，3H)，2.0(s，4H)，1.79(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，14H)。ESI-MS(m/z)1439[M+H] +。Ultimate analysis C 64H 94N 16O 20S theoretical value C 53.40, H, 6.58, N 15.57. measured value C 53.53, H 6.62, and N 15.41.
2) preparation of RGDVYIGSR
200mg (0.139mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 127mg (90%) target compound. 1H?NMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.80(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，2H)，5.0(s，lH)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，1H)，4.51(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，2H)，3.56(m，1H)，3.05(d，J＝6.0Hz，2H)，2.73(t，J＝6.0Hz，2H)，2.68(m，1H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，14H)。ESI-MS(m/z)1022[M+H] +。Ultimate analysis C 43H 71N 15O 14Theoretical value C 50.53, H, and 7.00, N20.56. measured value C 50.67, H 7.12, and N 20.42.
The preparation of embodiment 8RGDSYIGSR
1) Boc-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.295g (1.0mmol) Boc-Ser (Bzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add 0.780g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.93g (91%) target compound.ESI-MS(m/z)1021[M+H] +。
2) HClH-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
With 0.93g (0.9mmol) Boc-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)921[M+H] +。
3) Boc-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.291g (0.9mmol) Boc-Asp (Bzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.122g (0.9mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add 0.861g (0.9mmol) HClH-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.993g (91%) target compound.ESI-MS(m/z)1226[M+H] +。
4) HClH-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
With 0.993g (0.81mmol) Boc-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in l0ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)1126[M+H] +。
5) Boc-Arg (Tos)-Gly-Asp (OMe)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
With 0.305g (0.81mmol) Boc-Arg (NO 2)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.109g (0.81mmol) HOBt and 0.183g (0.89mmol) DCC down.Stir after 10 minutes, add 0.941g (0.81mmol) HClH-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.08g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.85(d，J＝5.0Hz，9H)，7.19(m，15H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，2H)，5.34(s，2H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，3H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，4H)，3.67(s，3H)，3.05(d，J＝6.0Hz，2H)，2.7(d，J＝7.0Hz，6H)，2.5(m，1H)，2.0(s，4H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，8H).ESI-MS(m/z)1484[M+H] +。Ultimate analysis C 68H 93N 17O 21Theoretical value C 55.01, H, and 6.31, N 16.04. measured value C 55.17, H 6.40, and N 16.21.
6) preparation of RGDSYIGSR
200mg (0.135mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 122mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝E11.0(s，2H)，7.80(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，2H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，2H)，4.50(t，J＝6.0Hz，2H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.56(m，1H)，3.05(d，J＝6.0Hz，2H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(Hs，10)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，8H)。SI-MS(m/z)1010[M+H] +。Ultimate analysis C 41H 67N 15O 15Theoretical value C48.75, H, 6.69, N 20.80. measured value C 48.88, H 6.81, and N 20.96.
The preparation of embodiment 9RGDFYIGSK
1) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-Tyr-Ile-Gly-Ser (Bzl)-Lyz (Z)-OBzl
0.762g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.916g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.46g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.85(d，J＝5.0Hz，10H)，7.87(d，J＝7.0Hz，2H)，7.34(d，J＝7.0Hz，2H)，7.19(m，20H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，1H)，5.34(s，4H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，3H)，4.63(s，2H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.67(s，3H)，3.05(d，J＝6.0Hz，4H)，2.96(q，J＝6.0Hz，2H)，2.7(d，J＝7.0Hz，4H)，2.5(m，1H)，2.35(s，3H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)1624[M+H] +。Ultimate analysis C 82H 105N 13O 20S theoretical value C 60.61, H, 6.51, N 11.21. measured value C 60.80, H 6.67, and N 11.38.
2) preparation of RGDFYIGSK
200mg (0.123mmol) Boc-Arg (TOS)-Gly-Asp (OMe)-Phe-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains product 114mg (89%). 1HNMR (BHSC-300, CDCl 3-d 3) δ=11.0 (s, 2H), 7.80 (d, J=5.0Hz, 8H), 7.21 (d, J=7.0Hz, 2H), 7.12 (m, 2H), 7.08 (t; J=7.0Hz, 1H), 6.95 (d, J=7.0Hz, 2H), 6.68 (d, J=7.0Hz, 2H), 5.7 (s, 1H), 5.0 (s; 1H), 4.92 (q, J=6.0Hz, 2H), 4.86 (q, J=6.0Hz, 1H), 4.62 (s, 1H), 4.50 (t, J=6.0Hz; 2H), 4.09 (d, J=6.0Hz, 4H), 4.03 (d, J=7.0Hz, 2H), 3.56 (m, 1H), 3.05 (d; J=6.0Hz, 4H), 2.73 (t, J=6.0Hz, 2H), 2.65 (q, J=6.0Hz, 4H), 2.5 (m, 1H); 2.0 (s, 8H), 1.79 (q, J=7.0Hz, 4H), 1.55 (m, 4H), 0.8-1.3 (m, J=8.0Hz, 10H).ESI-MS(m/z)1042[M+H] +。Ultimate analysis C 47H 71N 13O 14Theoretical value C 54.17, and H 6.87, N 17.47. measured value C 60.80, and H6.67, N 11.38.
The preparation of embodiment 10RGDVYIGSK
1) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Val-Tyr-Ile-Gly-Ser (Bzl)-Lyz (Z)-OBzl
0.713g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.916g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.40g (89%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.76(d，J＝5.0Hz，10H)，7.87(d，J＝7.0Hz，2H)，7.34(d，J＝7.0Hz，2H)，7.19(m，15H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，1H)，5.34(s，4H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，2H)，4.63(s，2H)，4.52(t，J＝6.0Hz，4H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.67(s，3H)，3.05(d，J＝6.0Hz，2H)，2.96(q，J＝6.0Hz，2H)，2.7(d，J＝7.0Hz，5H)，2.5(m，1H)，2.35(s，3H)，2.0(s，1H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，16H)。ESI-MS(m/z)1576[M+H] +。Ultimate analysis C 78H 105N 13O 20S theoretical value C 59.41, H, 6.71, N 11.55. measured value C 60.80, H 6.67, and N 11.38.
2) preparation of RGDSYIGSK
200mg (0.127mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 112mg (89%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.80(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，1H)，4.51(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，2H)，3.56(m，1H)，3.05(d，J＝6.0Hz，2H)，2.73(t，J＝6.0Hz，2H)，2.68(m，1H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，8H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，16H)。ESI-MS(m/z)995[M+H] +。Ultimate analysis C 43H 71N 13O 14Theoretical value C 51.95, H, and 7.20, N18.32. measured value C 52.11, H 7.29, and N 18.51.
The preparation of embodiment 11RGDSYIGSK
1) preparation of Boc-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
0.295g (1.0mmol) Boc-Ser (Bzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.916g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.054g (91%) target compound.ESI-MS(m/z)1159[M+H] +。
2) preparation of HClH-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
1.054g (0.9mmol) Boc-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in 10ml6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears; ETHYLE ACETATE is taken out in decompression, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)1059[M+H] +。
3) preparation of Boc-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
0.291g (0.9mmol) Boc-Asp (Bzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.122g (0.9mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add 0.985g (0.9mmol) HClNH 2The anhydrous THF solution of-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.113g (91%) target compound.ESI-MS(m/z)1363[M+H] +。
4) preparation of HClH-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
1.116g (0.81mmol) Boc-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution; Stirring at room 2 hours; TLC detects raw material point and disappears; ETHYLE ACETATE is taken out in decompression, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)1263[M+H] +。
5) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lyz (Z)-OBzl
With 0.305g (0.81mmol) Boc-Arg (NO 2)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.109g (0.81mmol) HOBt and 0.183g (0.89mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.941g (0.81mmol) HClH-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.18g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.75(d，J＝5.0Hz，10H)，7.19(m，25H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，1H)，5.34(s，6H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，3H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，4H)，3.05(d，J＝6.0Hz，2H)，2.96(q，J＝6.0Hz，2H)，2.7(d，J＝7.0Hz，4H)，2.5(m，1H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)1621[M+H] +。Ultimate analysis C 82H 104N 14O 21Theoretical value C 60.73, H, and 6.46, N 12.09. measured value C60.86, H 6.29, and N 12.25.
5) preparation of RGDSYIGSK
200mg (0.124mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 109mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.80(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，2H)，4.50(t，J＝6.0Hz，2H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.56(m，1H)，3.05(d，J＝6.0Hz，2H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)982[M+H] +。Ultimate analysis C 41H 67N 13O 15Theoretical value C 50.14, H, and 6.88, N 18.54. measured value C 50.27, H 6.97, and N 18.70.
The preparation of embodiment 12YIGSRRGDF
1) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-Arg (NO2)-Gly-Asp (OBzl)-PheOBzl
With 0.753g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.754g (1.0mmol) HClH-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.39g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.86(d，J＝5.0Hz，9H)，7.21(d，J＝7.0Hz，2H)，7.19(m，15H)，7.12(d，J＝7.0Hz，2H)，7.08(t，J＝7.0Hz，1H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，2H)，5.34(s，4H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.8(q，J＝5.0Hz，1H)，4.63(s，2H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.16(d，J＝6.0Hz，2H)，3.05(d，J＝6.0Hz，2H)，2.65(t，J＝7.0Hz，4H)，2.75(d，J＝7.0Hz，2H)，2.5(m，1H)，2.0(s，4H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，8H)。ESI-MS(m/z)1530[M+H] +。Ultimate analysis C 73H 95N 17O 20Theoretical value C 57.28, H, and 6.26, N 15.56. measured value C 57.13, H 6.17, and N 15.40.
2) preparation of YIGSRRGDF
With 200mg (0.131mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-Arg (NO2)-Gly-Asp-(OBzl)-Phe-OBzl is dissolved in the 4ml trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1ml methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100ml ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 125mg (90%) target compound.ESI-MS(m/z)1070[M+H] +。 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.89(d，J＝5.0Hz，8H)，7.21(d，J＝7.0Hz，2H)，7.12(m，2H)，7.08(t，J＝7.0Hz，1H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，2H)，5.0(s，1H)，4.86(q，J＝6.0Hz，2H)，4.62(s，1H)，4.51(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，2H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，8H)。Ultimate analysis C 47H 71N 15O 14Theoretical value C 52.75, and H 6.69, N 19.63. measured value C 52.89, and H 6.81, and N 19.47.
The preparation of embodiment 13YIGSKRRGDV
1) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-Arg (NO2)-Gly-Asp (OBzl)-Val-OBzl
With 0.753g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.706g (1.0mmol) HClH-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.35g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.86(d，J＝5.0Hz，9H)，7.19(m，15H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，2H)，5.34(s，4H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.63(s，2H)，4.52(t，J＝6.0Hz，3H)，4.41(t，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.09(m，1H)，3.05(d，J＝6.0Hz，2H)，2.65(t，J＝7.0Hz，4H)，2.75(d，J＝7.0Hz，2H)，2.5(m，1H)，2.0(s，4H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，14H)。ESI-MS(m/z)1482[M+H] +。Ultimate analysis C 69H 95N 17O 20Theoretical value C 55.90, H, and 6.46, N 16.06. measured value C 57.13, H 6.17, and N 15.40.
2) preparation of YIGSKRRGDV
200mg (0.135mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-Arg (NO 2)-Gly-Asp (OBzl)-Val-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 124mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.89(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，2H)，5.0(s，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，1H)，4.51(t，J＝6.0Hz，3H)，4.45(t，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，2H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，2H)，2.78(m，1H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，14H)。ESI-MS(m/z)1022[M+H] +。Ultimate analysis C 43H 71N 15O 14Theoretical value C 50.53, H, and 7.00, N 20.56. measured value C 50.36, H 7.10, and N 20.41.
The preparation of embodiment 14YIGSRRGDS
1) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-Arg (NO2)-Gly-Asp (OBzl)-Ser (Bzl) OBzl
With 0.753g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.784g (1.0mmol) HClH-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.42g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.86(d，J＝5.0Hz，9H)，7.19(m，20H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H，)，5.7(s，2H)，5.34(s，4H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.75(q，J＝5.0Hz，1H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.98(d，J＝6.0Hz，2H)，3.87(d，J＝5.0Hz，2H)，3.05(d，J＝6.0Hz，2H)，2.65(t，J＝7.0Hz，4H)，2.75(d，J＝7.0Hz，2H)，2.5(m，1H)，2.0(s，4H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，8H)。ESI-MS(m/z)1596[M+H] +。Ultimate analysis C 77H 97N 17O 21Theoretical value C 57.92, H, and 6.12, N 14.91. measured value C 57.80, H 6.03, N14.78.
2) preparation of YIGSRRGDS
With 200mg (0.128mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-Arg (NO 2)-Gly-Asp-(OBzl)-Ser (Bzl)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 116mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.89(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，2H)，5.0(s，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，1H)，4.53(t，J＝6.0Hz，4H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，2H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，10H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，8H)。ESI-MS(m/z)1011[M+H] +。Ultimate analysis C 41H 67N 15O 15Theoretical value C 48.75, H, and 6.69, N 20.80. measured value C48.61, H 6.57, and N 20.96.
The preparation of embodiment 15YIGSKRGDF
1) preparation of Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Arg (NO2)-Gly-Asp (OBzl)-PheOBzl
0.890g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.754g (1.0mmol) HClH-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.46g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.86(d，J＝5.0Hz，10H)，7.21(d，J＝7.0Hz，2H)，7.19(m，20H)，7.12(d，J＝7.0Hz，2H)，7.08(t，J＝7.0Hz，1H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，1H)，5.34(s，6H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.8(q，J＝5.0Hz，1H)，4.63(s，2H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.16(d，J＝6.0Hz，2H)，3.05(d，J＝6.0Hz，2H)，2.96(t，J＝7.0Hz，2H)，2.65(t，J＝7.0Hz，2H)，2.75(d，J＝7.0Hz，2H)，2.5(m，1H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)1591[M+H] +。Ultimate analysis C 77H 97N 17O 21Theoretical value C 61.12, H, and 6.46, N 12.32. measured value C 61.00, H 6.38, and N 12.50.
2) preparation of YIGSKRGDF
200mg (0.124mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-Arg (NO 2)-Gly-Asp (OBzl)-Phe-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 116mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.89(d，J＝5.0Hz，8H)，7.21(d，J＝7.0Hz，2H)，7.12(m，2H)，7.08(t，J＝7.0Hz，1H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，1H)，5.0(s，1H)，4.86(q，J＝6.0Hz，2H)，4.62(s，1H)，4.51(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，2H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.73(t，＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，8H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)1042[M+H] +。Ultimate analysis C 47H 71N 13O 14Theoretical value C 54.17, H, and 6.87, N 17.47. measured value C 54.31, H 6.96, and N 17.61.
The preparation of embodiment 16YIGSKRGDV
1) preparation of Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Arg (NO2)-Gly-Asp (OBzl)-Val-OBzl
0.890g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.706g (1.0mmol) HClH-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.42g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.86(d，J＝5.0Hz，10H)，7.19(m，20H)，6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，1H)，5.34(s，6H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.63(s，2H)，4.52(t，J＝6.0Hz，3H)，4.41(t，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，2H)，3.09(m，1H)，3.05(d，J＝6.0Hz，2H)，2.96(t，J＝7.0Hz，2H)，2.65(t，J＝7.0Hz，2H)，2.75(d，J＝7.0Hz，2H)，2.5(m，1H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，16H)。ESI-MS(m/z)1543[M+H] +。Ultimate analysis C 77H 102N 14O 20Theoretical value C 59.91, H, and 6.66, N 12.70. measured value C 60.07, H 6.79, and N 12.53.
2) preparation of YIGSKRGDV
With 200mg (0.128mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Arg (NO 2)-Gly-Asp (OBzl)-Val-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 115mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.89(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，1H)，5.0(s，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，1H)，4.51(t，J＝6.0Hz，3H)，4.45(t，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，2H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，2H)，2.78(m，1H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，8H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，16H)。ESI-MS(m/z)994[M+H] +。Ultimate analysis C 43H 71N 13O 14Theoretical value C 51.95, H, and 7.20, N 18.32. measured value C 52.18, H 7.33, and N 18.20.
The preparation of embodiment 17YIGSKRGDS
1) preparation of Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Arg (NO2)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl
0.890g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.784g (1.0mmol) HClH-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.49g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.86(d，J＝5.0Hz，10H)，7.19(，m，25H)6.95(d，J＝7.0Hz，2H)，6.65(d，J＝7.0Hz，2H)，5.7(s，1H)，5.34(s，6H)，5.17(q，J＝6.0Hz，1H)，5.0(s，1H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，1H)，4.75(q，J＝5.0Hz，1H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.09(d，J＝6.0Hz，4H)，3.98(d，J＝6.0Hz，2H)，3.87(d，J＝5.0Hz，2H)，3.05(d，J＝6.0Hz，2H)，2.96(t，J＝7.0Hz，2H)，2.65(t，J＝7.0Hz，2H)，2.75(d，J＝7.0Hz，2H)，2.5(m，1H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.4(s，9H)，1.45-1.6(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)1595[M+H] +。Ultimate analysis C 77H 97N 17O 21Theoretical value C 57.92, H, and 6.12, N 14.91. measured value C 58.08, H 6.03, and N 14.77.
2) preparation of YIGSKRGDS
With 200mg (0.122mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Arg (NO 2)-Gly-Asp-(OBzl)-Val-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 107mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，2H)，7.89(d，J＝5.0Hz，8H)，6.95(d，J＝7.0Hz，2H)，6.68(d，J＝7.0Hz，2H)，5.7(s，1H)，5.0(s，1H)，4.86(q，J＝6.0Hz，1H)，4.62(s，1H)，4.53(t，J＝6.0Hz，4H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，2H)，2.73(t，J＝6.0Hz，2H)，2.65(q，J＝6.0Hz，4H)，2.5(m，1H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，10H)。ESI-MS(m/z)982[M+H] +。Ultimate analysis C 41H 67N 13O 15Theoretical value C 50.14, H, and 6.88, N 18.54. measured value C 50.29, H 6.77, and N 18.69.
The preparation of embodiment 18RGDFRGDF
1) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-Arg (Tos)-Gly-Asp (OMe)-Phe-OBzl
0.761g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.787g (1.0mmol) HClH-Arg (Tos)-Gly-Asp (OMe)-Phe-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.36g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.89(d，J＝5.0Hz，8H)，7.81(d，J＝7.0Hz，4H)，7.19(m，5H)，7.34(d，J＝7.0Hz，4H)，7.21(d，J＝7.0Hz，4H)，7.12(d，J＝7.0Hz，4H)，7.08(t，J＝7.0Hz，2H)，5.7(s，2H)，5.34(s，2H)，5.17(q，J＝6.0Hz，2H)，4.92(q，J＝6.0Hz，1H)，4.81(d，J＝6.0Hz，1H)，4.53(q，J＝6.0Hz，2H)，4.09(d，J＝6.0Hz，4H)，3.67(s，6H)，3.56(q，J＝6.0Hz，1H)，3.16(d，J＝6.0Hz，2H)，3.05(d，J＝6.0Hz，2H)，2.75(d，J＝7.0Hz，4H)，2.65(t，J＝7.0Hz，4H)，2.35(s，6H)，2.0(s，4H)，1.79(q，J＝7.0Hz，4H)，1.4(s，9H)，1.56(m，4H)。ESI-MS(m/z)1495[M+H] +。Ultimate analysis C 70H 90N 14O 19S 2Theoretical value C 56.21, H, and 6.07, N 13011. measured value C 56.09, H 6.14, and N 13.00.
2) preparation of RGDFRGDF
200mg (0.132mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Phe-Arg (Tos)-Gly-Asp-(OMe)-Phe-OBzl is dissolved in the 4ml trifluoracetic acid; Add 1ml trifluoromethanesulfonic acid and 1ml methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and inferior property adding 100ml ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 115mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，3H)，7.89(d，J＝5.0Hz，7H)，7.21(d，J＝7.0Hz，4H)，7.12(m，4H)，7.08(t，J＝7.0Hz，2H)，5.7(s，2H)，4.92(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，3H)，4.53(t，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.56(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.73(t，J＝6.0Hz，4H)，2.65(q，J＝6.0Hz，4H)，2.0(s，8H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)。ESI-MS(m/z)969[M+H] +。Ultimate analysis C 42H 60N 14O 13Theoretical value C 52.06, H, and 6.24, N20.24. measured value C 52.22, H 6.33, and N 20.41.
The preparation of embodiment 19RGDVRGDV
1) preparation of Boc-Arg (Tos)-Gly-Asp (OMe)-Val-Arg (Tos)-Gly-Asp (OMe)-Val-OBzl
0.713g (1.0mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.740g (1.0mmol) HClH-Boc-Arg (Tos)-Gly-Asp (OMe)-Val-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.29g (91%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.89(d，J＝5.0Hz，8H)，7.81(d，J＝7.0Hz，4H)，7.19(m，5H)，7.34(d，J＝7.0Hz，4H)，5.7(s，2H)，5.34(s，2H)，5.17(q，J＝6.0Hz，2H)，4.53(q，J＝6.0Hz，1H)，4.52(t，J＝6.0Hz，2H)，4.09(d，J＝6.0Hz，4H)，3.67(s，6H)，3.56(q，J＝6.0Hz，1H)，2.75(d，J＝7.0Hz，4H)，2.68(m，J＝8.0Hz，2H)2.65(t，J＝7.0Hz，4H)，2.35(s，6H)，2.0(s，4H)，1.79(q，J＝7.0Hz，4H)，1.4(s，9H)，1.56(m，4H)，0.98(d，J＝8.0Hz，12H)。ESI-MS(m/z)1399[M+H] +。Ultimate analysis C 62H 90N 14O 19S 2Theoretical value C 53.21, H, and 6.48, N 14.01. measured value C 53.37, H 6.60, N14.18.
2) preparation of RGDVRGDV
200mg (0.141mmol) Boc-Arg (Tos)-Gly-Asp (OMe)-Val-Arg (Tos)-Gly-Asp-(OMe)-Val-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 111mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，3H)，7.89(d，J＝5.0Hz，7H)，5.7(s，2H)，4.86(q，J＝6.0Hz，2H)，4.53(t，J＝6.0Hz，2H)，4.45(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.56(q，J＝6.0Hz，1H)，2.78(m，1H)，2.73(t，J＝6.0Hz，4H)，2.68(m，1H)，2.65(q，J＝6.0Hz，4H)，2.0(s，8H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，1.01(d，J＝8.0Hz，12H)。ESI-MS(m/z)873[M+H] +。Ultimate analysis C 34H 60N 14O 13Theoretical value C 46.78, and H 6.93, N 22.46. measured value C 46.62, and H 6.87, and N 22.62.
The preparation of embodiment 20RGDSRGDS
1) Boc-Ser (Bzl)-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Ser-OBzl
0.295g (1.0mmol) Boc-Ser (Bzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.784g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 0.933g (91%) target compound.ESI-MS(m/z)1026[M+H] +。
2) HClH-Ser (Bzl)-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Ser-OBzl
With 0.933g (0.9mmol) Boc-Ser (Bzl)-Arg (NO 2)-Gly-Asp (OBzl)-Ser-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)926[M+H] +。
3) preparation of Boc-Asp (OBzl)-Ser (Bzl)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
0.291g (0.9mmol) Boc-Asp (Bzl)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.122g (0.9mmol) HOBt and 0.23g (1.09mmol) DCC down.Stir after 10 minutes, add 0.865g (0.9mmol) HClH-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Ser-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.02g (91%) target compound.ESI-MS(m/z)1248[M+H] +。
4) HClH-Asp (OBzl)-Ser (Bzl)-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Ser-OBzl
With 1.02g (0.81mmol) Boc-Asp (OBzl)-Ser (Bzl)-Arg (NO 2)-Gly-Asp (OBzl)-Ser-OBzl is dissolved in 10ml 6mol/l hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, and TLC detects raw material point and disappears, and reduces pressure and takes out ETHYLE ACETATE, adds a small amount of ether repeatedly and reduces pressure and bleed to remove the acid gas in the product.Add a small amount of ether at last product is ground to form pressed powder, directly be used for next step reaction.ESI-MS(m/z)1148[M+H] +。
5) Boc-Arg (NO 2)-Gly-Asp (OBzl) Ser (Bzl)-Arg (NO 2The preparation of)-Gly-Asp (OBzl)-Ser (Bzl)-OBzl
With 0.305g (0.81mmol) Boc-Arg (NO 2)-Gly-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.109g (0.81mmol) HOBt and 0.183g (0.89mmol) DCC down.Stir after 10 minutes, add 0.885g (0.81mmol) HClH-Asp (OBzl) Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-Gly-Asp (OBzl)-Ser-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.10g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.89(d，J＝5.0Hz，8H)，7.19(m，25H)，5.7(s，2H)，5.34(s，6H)，5.17(q，J＝6.0Hz，2H)，4.86(q，J＝6.0Hz，1H)，4.63(s，4H)，4.75(q，J＝6.0Hz，1H)，4.53(q，J＝6.0Hz，2H)，4.09(d，J＝6.0Hz，4H)，3.98(d，J＝7.0Hz，2H)，3.87(d，J＝7.0Hz，2H)，2.75(d，J＝7.0Hz，4H)，2.65(q，J＝7.0Hz，4H)，2.0(s，4H)，1.79(q，J＝7.0Hz，4H)，1.4(s，9H)，1.55(m，4H)。ESI-MS(m/z)1348[M+H] +。Ultimate analysis C 70H 88N 6O 21Theoretical value C 62.30, H, and 6.57, N 6.23. measured value C 62.42, H 6.68, and N 6.10.
6) preparation of RGDSRGDS
With 200mg (0.133mmol) Boc-Arg (NO 2)-Gly-Asp (OBzl)-Ser (Bzl)-Arg (NO 2)-Gly-Asp-(OBzl)-Ser (Bzl)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 103mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，3H)，7.89(d，J＝5.0Hz，7H)，5.7(s，2H)，4.62(q，J＝6.0Hz，1H)，4.86(q，J＝6.0Hz，2H)，4.55(q，J＝6.0Hz，1H)，4.53(t，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.56(q，J＝6.0Hz，1H)，2.73(t，J＝6.0Hz，4H)，2.65(q，J＝6.0Hz，4H)，2.0(s，10H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)。ESI-MS(m/z)849[M+H] +。Ultimate analysis C 30H 52N 14O 15Theoretical value C 42.45, and H 6.17, N 23.10. measured value C 42.29, and H 6.09, and N 23.27.
The preparation of embodiment 21YIGSKYIGSK
1) preparation of Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
0.890g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.916g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.57g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.88(d，J＝5.0Hz，12H)，7.19(m，25H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.34(s，6H)，5.0(s，2H)，4.92(q，J＝6.0Hz，2H)，4.86(q，J＝6.0Hz，2H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.42(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，4H)，3.05(d，J＝6.0Hz，4H)，2.96(q，J＝6.0Hz，4H)，2.5(m，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.55(m，4H)，1.4(s，9H)，0.8-1.3(m，J＝8.0Hz，20H)。ESI-MS(m/z)1753[M+H] +。Ultimate analysis C 70H 88N 6O 21Theoretical value C 64.37, H, and 6.90, N 9.58. measured value C 64.49, H 6.81, and N 9.74.
2) preparation of YIGSKYIGSK
200mg (0.114mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 114mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，1H)，7.89(d，J＝5.0Hz，9H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.0(s，2H)，4.92(q，J＝6.0Hz，1H)，4.62(q，J＝6.0Hz，2H)，4.53(t，J＝6.0Hz，3H)，4.46(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.65(q，J＝6.0Hz，4H)，2.5(m，2H)，2.0(s，8H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，20H)。ESI-MS(m/z)1115[M+H] +。Ultimate analysis C 52H 82N 12O 15Theoretical value C 56.00, H, and 7.41, N 15.07. measured value C 56.14.49, H 7.52, and N 15.20.
The preparation of embodiment 22YIGSKYIGSR
1) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
0.890g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.800g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.45g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.88(d，J＝5.0Hz，11H)，7.19(m，15H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.7(s，1H)，5.34(s，2H)，5.0(s，2H)，4.92(q，J＝6.0Hz，2H)，4.86(q，J＝6.0Hz，2H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.42(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，4H)，3.67(s，3H)，3.05(d，J＝6.0Hz，4H)，2.96(q，J＝6.0Hz，2H)，2.65(t，J＝7.0Hz，2H)，2.5(m，2H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.55(m，4H)，1.4(s，9H)，0.8-1.3(m，J＝8.0Hz，18H)。ESI-MS(m/z)1616[M+H] +。Ultimate analysis C 80H 109N 15O 21Theoretical value C 59.43, H, and 6.80, N 12.99. measured value C59.61, H 6.92, and N 12.83.
2) preparation of YIGSKYIGSR
With 200mg (0.124mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 127mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，1H)，7.89(d，J＝5.0Hz，9H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.7(s，1H)，5.0(s，2H)，4.92(q，J＝6.0Hz，1H)，4.62(q，J＝6.0Hz，2H)，4.53(t，J＝6.0Hz，3H)，4.46(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.65(q，J＝6.0Hz，4H)，2.5(m，2H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，18H)。ESI-MS(m/z)1144[M+H] +。Ultimate analysis C 52H 82N 14O 15Theoretical value C 54.63, H, and 7.23, N 17.15. measured value C 54.50, H 7.09, and N 17.34.
The preparation of embodiment 23YIGSRYIGSK
1) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl
With 0.829g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add the anhydrous THF solution of 0.916g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-OBzl.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.52g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.88(d，J＝5.0Hz，11H)，7.19(m，20H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.7(s，1H)，5.34(s，4H)，5.0(s，2H)，4.92(q，J＝6.0Hz，2H)，4.86(q，J＝6.0Hz，2H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.42(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，4H)，3.05(d，J＝6.0Hz，4H)，2.96(q，J＝6.0Hz，2H)，2.65(t，J＝7.0Hz，2H)，2.5(m，2H)，2.0(s，2H)，1.7-1.9(q，J＝7.0Hz，4H)，1.55(m，4H)，1.4(s，9H)，0.8-1.3(m，J＝8.0Hz，18H)。ESI-MS(m/z)1693[M+H] +。Ultimate analysis C 86H 113N 15O 21Theoretical value C 61.01, H, and 6.73, N 12.41. measured value C60.86, H 6.80, and N 12.57.
2) preparation of YIGSRYIGSK
200mg (0.118mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Tyr-Ile-Gly-Ser (Bzl)-Arg-(NO2)-OMe is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 10 () mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 121mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，1H)，7.89(d，J＝5.0Hz，9H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.7(s，1H)，5.0(s，2H)，4.92(q，J＝6.0Hz，1H)，4.62(q，J＝6.0Hz，2H)，4.53(t，J＝6.0Hz，3H)，4.46(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.65(q，J＝6.0Hz，4H)，2.5(m，2H)，2.0(s，9H)，1.79(q，J＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，18H)。ESI-MS(m/z)1172[M+H] +。Ultimate analysis C 52H 82N 16O 15Theoretical value C 53.32, and H 7.06, N 19.13. measured value C 53.44, and H 7.17, and N 19.00.
The preparation of embodiment 24YIGSRYIGSR
1) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The preparation of)-OMe
With 0.829g (1.0mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OH is dissolved among the anhydrous THF of 5ml, and 0 ℃ adds 0.135g (1.0mmol) HOBt and 0.23g (1.1mmol) DCC down.Stir after 10 minutes, add 0.780g (1.0mmol) HClH-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2The anhydrous THF solution of)-OMe.Reaction mixture is regulated pH 8-9 with NMM, and 0 ℃ was stirred 24 hours.Stopped reaction removes by filter the DCU that is settled out.Filtrate decompression concentrates, and residue is dissolved in the ETHYLE ACETATE, and the solution that obtains is used saturated NaHCO successively 3The aqueous solution, 5%KHSO 4The aqueous solution and the saturated NaCl aqueous solution are washed.Ethyl acetate layer is used anhydrous Na SO 4Dry 3 hours.Filtering NaSO 4, filtrate decompression concentrates removes ETHYLE ACETATE.Residue uses column chromatography (CHCl 3: CH 3OH, 30: 1).Obtain 1.40g (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝7.88(d，J＝5.0Hz，10H)，7.19(m，10H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.7(s，2H)，5.0(s，2H)，4.92(q，J＝6.0Hz，2H)，4.86(q，J＝6.0Hz，2H)，4.63(s，4H)，4.52(t，J＝6.0Hz，3H)，4.42(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，3.87(d，J＝5.0Hz，4H)，3.67(s，3H)，3.05(d，J＝6.0Hz，4H)，2.65(t，J＝7.0Hz，4H)，2.5(m，2H)，2.0(s，4H)，1.7-1.9(q，J＝7.0Hz，4H)，1.55(m，4H)，1.4(s，9H)，0.8-1.3(m，J＝8.0Hz，16H)。ESI-MS(m/z)1556[M+H] +。Ultimate analysis C 72H 102N 18O 21Theoretical value C 55.59, H, and 6.61, N 16.21. measured value C55.72, H 6.71, and N 16.09.
2) preparation of YIGSRYIGSR
With 200mg (0.129mmol) Boc-Tyr-Ile-Gly-Ser (Bzl)-Lys (Z)-Tyr-Ile-Gly-Ser (Bzl)-Arg (NO 2)-OMe is dissolved in the 4mL trifluoracetic acid; Add 1mL trifluoromethanesulfonic acid and 1mL methyl-phenoxide; Stirring reaction is 1 hour under 0 ℃ of condition of ice bath, and disposable adding 100mL ether was bled in decompression 5 minutes; The product pressed powder of separating out uses Sephadex G 10 to purify, and obtains 135mg (90%) target compound. 1HNMR(BHSC-300，CDCl 3-d 3)δ＝11.0(s，1H)，7.89(d，J＝5.0Hz，9H)，6.95(d，J＝7.0Hz，4H)，6.68(d，J＝7.0Hz，4H)，5.7(s，2H)，5.0(s，2H)，4.92(q，J＝6.0Hz，1H)，4.62(q，J＝6.0Hz，2H)，4.53(t，J＝6.0Hz，3H)，4.46(q，J＝6.0Hz，1H)，4.09(d，J＝6.0Hz，4H)，4.03(d，J＝7.0Hz，4H)，3.95(q，J＝6.0Hz，1H)，3.05(d，J＝6.0Hz，4H)，2.65(q，J＝6.0Hz，4H)，2.5(m，2H)，2.0(s，10H)，1.79(q，＝7.0Hz，4H)，1.55(m，4H)，0.8-1.3(m，J＝8.0Hz，16H)。ESI-MS(m/z)1172[M+H] +。Ultimate analysis C 52H 82N 16O 15Theoretical value C 53.32, and H 7.06, N 19.13. measured value C 53.19, and H 7.15, and N 19.00.
Test Example 1 polypeptide antithrombotic acitivity test of the present invention
1) rat operation and apparatus
(male, 220～230g) press 1200mgkg to the SD rat -1Dosage abdominal injection urethane solution is anaesthetized.The anesthetized rat dorsal position is fixed, and separates RCCA, and in proximal part folder bulldog clamp, proximal part and distal end penetrate surgical thread respectively, and the surgical thread of distal end is clamped with mosquito forceps in fur, and preparation is in the distal end intubate.
2) intubate.
Intubate is the polyethylene rubber tube that silylanization is crossed, and divides three sections, and the stage casing is a polyethylene rubber tube, long 60.0mm, internal diameter 3.5mm; Two ends are identical polyethylene tube, and pipe range 100.0mm, the end that internal diameter 1.0mm, external diameter 2.0mm should manage pull into point pipe (being used to insert rat carotid artery or vein), and external diameter is 1.0mm.Be respectively charged into the long black surgical thread of 6cm in the clean penicillium mould bottle with the number of finishing, weigh; Take out silk thread then, put into the thicker intubate in stage casing of ready intubate according to numbering.
Open rat right side bulldog clamp, fill with heparin-saline solution (50IUkg in will managing through sharp pipe end with syringe -1), then the arterial end of intubate is inserted the rat right carotid, the heparin of calculated amount is slowly injected in the rat body.
3) administration.
Medicine: saline water (3mlkg -1), the physiological salt soln of the physiological salt soln of Asprin (dosage is 30mg/kg), compound of the present invention (dosage is 10 μ mol/kg).
Folder closes rat right carotid artery folder, pulls up the syringe of intubate vein end, has the water-soluble medical fluid injector of medicine of calculated amount to insert the vein end of intubate suction, opens rat right carotid artery folder, and medicine is slowly pushed in the rat body.Folder closes right carotid artery folder, and the vein end in teahouse is inserted the rats with left jugular vein of separator well, opens bulldog clamp, makes blood begin circulation.And pick up counting simultaneously.Can produce thrombus because of blood circulation on the silk thread in this process in the extra heavy pipe of intubate central authorities.
4) thrombus is weighed.
Timing is cut off venous incubation after beginning 15 minutes, stops circulation, carefully takes out silk thread with the ophthalmology tweezer, on filter paper, dips in drop of blood gently, puts into the penicillium mould bottle of weighing in advance, accurately weighs and record.Calculate the weight in wet base of thrombus.Each medicine repeats 11 administrations.The wet weight of thrombus of each group of statistics (X ± SD), and do the t check.
5) result
Compound of the present invention all has good anti-thrombus activity.The result lists table 1 in.
The antithrombotic acitivity of table 1 compound of the present invention on rat model
Figure G200610144234920061218D000401
N=11; NS=saline water; A. compare P＜0.001 with saline water.

Claims (3)

1. have the polypeptide of target antithrombotic activity, it is characterized in that, this polypeptide is following aminoacid sequence: YIGSRY IGSR.
2. method for preparing claim 1 polypeptide comprises:
(1), adopts liquid-phase synthesis process, through progressively connecing the protection midbody of the synthetic YIGSR of peptide;
(2), slough the N end protection base of the protection midbody of YIGSR;
(3), slough the C end protection base of the protection midbody of YIGSR;
(4), put together formula to the YIGSR of the YIGSR of N end dissociative protection midbody and C end dissociative protection midbody and modify, slough again and put together formula and modify and obtain the YIGSRYIGSR polypeptide.
3. the polypeptide of claim 1 is in the purposes of preparation in the antithrombotic reagent.
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CN2012101050827A Division CN102702312A (en) 2006-11-30 2006-11-30 RGDVYIGSK with targeting antithrombotic activity, preparation and applications thereof
CN2012101051389A Division CN102702317A (en) 2006-11-30 2006-11-30 RGDSYIGSR with target antithrombotic activity, preparation and application thereof
CN2012101050850A Division CN102702313A (en) 2006-11-30 2006-11-30 RGDV-YIGSR with targeted anti-thrombotic activity, preparation and application thereof
CN2012101051228A Division CN102702315A (en) 2006-11-30 2006-11-30 RGDSYIGSK with targeting antithrombotic activity, preparation and applications thereof
CN2012101051478A Division CN102702318A (en) 2006-11-30 2006-11-30 YIGSKYIGSK with target antithrombotic activity, and preparation and application thereof
CN2012101051251A Division CN102702316A (en) 2006-11-30 2006-11-30 RGDFYIGSK with targeting antithrombotic activity, preparation and applications thereof
CN2012101051073A Division CN102702314A (en) 2006-11-30 2006-11-30 RGDF-YIGSR with targeted anti-thrombotic activity, preparation and application thereof

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