CN101492494A - Integrin ligand cyclic peptide analogues and cyclization method - Google Patents
Integrin ligand cyclic peptide analogues and cyclization method Download PDFInfo
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Abstract
The invention relates to a cyclopeptide analog and a cyclization method thereof, which aims at providing an integrin ligand cyclopeptide analog and a cyclization method thereof. The cyclization method comprises the following steps: Azido-glycine is firstly generated by glycin with the effects of trifluoromethanesulfonic anhydride and Sodium azide; Fmoc-Asp-Propargyl is then generated by condensation with propargylamine in solution; after side-chain carboxyl of propargyl acyl aspartic acid is then hung on 2-CTC resin, linear tetrapeptide analogs are condensed in turn, and the linear tetrapeptide analogs are then cut down from the resin; and the cyclopeptide analog is synthesized in the condition of liquid phase (DCM is used as a solvent and CuBr/DBU is used as catalyst) after 4 to 6 hours of reaction at room temperature. The cyclopeptide analog and cyclization method has the following advantages: (1) the integrin ligand cyclopeptide analog in the invention is introduced with heterocyclic rings (triazole rings), thereby being capable of increasing biological activity; and (2) the cyclization method in the invention is more simple, convenient and high-efficient, and the condition is moderate.
Description
Technical field
The present invention relates to cyclic peptide analogue and cyclization method thereof, in particular, relate to integrin ligand cyclic peptide analogues and cyclization method thereof.
Background technology
An evident characteristic of tumor growth is by stimulating vascular endothelial cell to secrete various somatomedins, further promote tumor vessel to grow fast, making cell enough blood supplies be arranged and malignancy.Growth of tumor and transfer are processes that depends on blood vessel, can obtain needed nutrition by the disperse of tumour surrounding blood vessel when diameter of tumor is less than 2～3mm usually.Tumour increases, and then needs the formation of new vessel to supply with its nutrition.As far back as 1971, U.S. scientist Folkman etc. proposed and can suppress growth of tumor by the tumor vascular generation of blocking-up, prevent the transfer of tumour.According to this hypothesis, the researchist wishes by the blood supply of cutting off blood vessel, breaking off tumour growth of tumor to be obstructed, and reaches the purpose that stops the tumor vessel overgrowing, and finally makes the hungry and death of cancer cells " wean ".
To integrate plain (integrin) be a class by α subunit and β subunit through the heterodimer transmembrane glycoprotein adhesion molecule that non covalent bond is formed by connecting, and comprises C-terminal in the short tenuigenin, strides film section and long extracellular N-terminal.It has high expression level on kinds of tumor cells surface and new vessel endotheliocyte, but in mature blood endothelial cell and most normal organ system, the integration element is not expressed or very a spot of expression.Present known have at least 14 kinds of α subunits and 9 kinds of β subunits, kind of hypotype surplus the integration element of composition has reached 20.At intracellular region, interactions plain and cytoskeletal protein and other tenuigenin compositions are integrated in the mediation of β chain.Integrate element and have two kinds of major functions: a kind of is by combining the adhesion of mediated cell and basilar membrane, cell and cell with respective ligand, and this is that it mainly acts on; Another kind is the bridge that transmits as external information in the cell, and on the one hand intracellular Signal Regulation is integrated avidity and active condition plain and its part, integrates the plain back that combines with its part on the other hand and transmits signal in cell, the performance biological effect.Integrin receptor by with acceptor recognition sequence RGD (arginine-glycine-aspartic acid) specific combination of extracellular matrix protein (as Vitronectins etc.); mediation tumor cell adhesion and migration; in the angiogenesis of tumor growth, local infiltration, transfer, particularly tumor inducing, play a significant role.
Above-mentioned result of study for the design contain the RGD sequence integrin receptor small molecules antagonism peptide, as selectivity cancer target developer or the treatment preparation established sufficient theoretical basis.Therefore, in the more than ten years in the past, many seminars (Haubner etal.J.Am.Chem.Soc.118:7461-7472,1996; Hammes et al.Nature Med.2:529-533,1996; Wermuth et al.J.Am.Chem.Soc.199:1328-1335,1997; Dechantsreiter et al.Med.Chem.42:3033-3040,1999; ) successively design a series of RGD of containing polypeptide of sequence, wherein more representational is the design synthetic cRGDfK of Kessler seminar, cRGDfV and methylated analogue cRGDfV (Me) thereof are (Figure1.).With cRGDfK is example, and the method that they adopt is to synthesize earlier linearity on the Fmoc resin, and the RGDfK of side chain protected scales off cyclisation with linear peptides from resin then, removes the side chain protected group at last.Because during cyclisation is common amido linkage condensation reaction, the cyclisation productive rate greatly reduces than common amido linkage condensation.In addition; sometimes also need some deleterious malodorous reagent; as diphenyl phosphoryl azide (diphenylphosphoryl; DPPA); thioanisole (thioanisole), dithioglycol (ethanedithiol), therefore; need to seek a kind of reaction conditions gentleness, the cyclization method that productive rate is higher.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a kind of novel integrin ligand cyclic peptide analogues is provided, further object of the present invention provides the cyclization method of this integrin ligand cyclic peptide analogues.
In order to overcome the above problems, the present invention is achieved through the following technical solutions.
The invention provides a kind of integrin ligand cyclic peptide analogues, its structure is as follows:
The present invention also provides the cyclization method of aforementioned integrin ligand cyclic peptide analogues, may further comprise the steps: (1) hangs over fluorenylmethyloxycarbonyl-alkynes propionyl-aspartic acid (Fmoc-Asp-propargyl) on the 2-chlorine trityl chloride resin (2-CTC resin), the tetrapeptide analogs of synthesizing linear:
Take by weighing 1g 2-CTC resin in solid phase reactor, (DCM) 15ml swelling that adds methylene chloride, washing resin; In the good resin of swelling, add Fmoc-Asp-propargyl and N, N '-diisopropylethylamine (DIPEA), reaction is 2.5 hours under the room temperature, with the ethyl acetate (EA) of 3 * 20ml and the water washing of 10ml; Piperidines/dimethylformamide with 20% (DMF) solution removes fluorenylmethyloxycarbonyl (Fmoc), reacts 20 minutes; Water washing with EA and the 10ml of 3 * 20ml, add fluorenylmethyloxycarbonyl-glycine (Fmoc-Gly-OH), benzotriazole-N, N, N ', N '-tetramethyl-urea phosphofluoric acid ester (HBTU) and N-methylmorpholine (NMM) are to reactor, reacted 2 hours, with triketohydrindene hydrate indicator detection reaction, with the water washing of EA and the 10ml of 3 * 20ml; Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes; Water washing with EA and the 10ml of 3 * 20ml;
Adopt the identical operations step; condensation fluorenylmethyloxycarbonyl-2,2 successively, 4; 6,7-pentamethyl-benzo furans-5-alkylsulfonyl-arginine [Fmoc-Arg (Pbf)-OH], Fmoc-Xaa-OH and nitrine glycine (Azido-glycine) generate linear tetrapeptide analogs.
(2) cyclisation:
The tetrapeptide analogs of the full guard of linearity is scaled off from resin, and with 2: 8 trifluoroacetic acid/dichloromethane of lysate (TFA/DCM) and several acetate (AcOH), cracking 2 hours is spin-dried for lysate; Make solvent at DCM; cuprous bromide/1; 8-diazacyclo ring (5,4,0) 7-hendecene (CuBr/DBU) is done to react 4～6 hours under the liquid-phase condition of catalyzer and the room temperature; revolve DCM after the reaction; add lysate and remove side chain protected cracking 2 hours, revolve lysate, ether sedimentation on the rocks; the centrifugal supernatant liquor of removing obtains solid, is the final product integrin ligand cyclic peptide.
As a kind of improvement, in order to use Click Chemistry method, described Fmoc-Asp-propargyl makes in the following manner:
Fluorenylmethyloxycarbonyl-aspartic acid-beta-tert-butyl ester [Fmoc-Asp (OtBu)-OH] of 0.93g is dissolved among tetrahydrofuran (THF) (THF) 25ml, add 2.49g N then successively, the 1-hydroxy benzo triazole (HOBt) of N '-dicyclohexylcarbodiimide (DCC) and 1.62g; Propargylamine (0.165g) adds in the reactor after with 5ml THF mixing, reaction 2h, and with triketohydrindene hydrate indicator detection reaction, reaction product is with the water washing of EA and the 10ml of 3 * 20ml; With volume ratio is that 2: 3 TFA/DCM removes side chain carboxyl group blocking group OtBu, with the water washing of EA and the 10ml of 3 * 20ml; Rotary evaporation steams solvent, and vacuum-drying obtains white solid, i.e. Fmoc-Asp-propargyl.
As a kind of improvement, in order to use Click Chemistry method, described nitrine glycine (Azido-glycine) makes in the following manner:
(1) with 6g sodium azide (NaN
3) be dissolved in the round-bottomed flask of 150ml, adding the DCM of 24ml again, ice bath is cooled to 0 ℃, stirs 20 minutes; Reaction mixture extracts with the DCM of 2 * 12ml; Merge organic phase, with saturated sodium carbonate (NaHCO
3) wash, promptly obtain being dissolved in the fluoroform sulfonyl azide of DCM, need not to be further purified next step reaction of direct adding and use;
(2) with the salt of wormwood (K of 0.675g glycine, 1.783g
2CO
3), the copper sulfate (CuSO of 0.0226g
4) join in the round-bottomed flask, add the DCM that 30ml water, 60ml methyl alcohol and 48ml contain the fluoroform sulfonyl azide again, at room temperature stir, reacted 24 hours; The rotary evaporation that subsequently reaction mixture reduced pressure is removed organic solvent, and remaining part is used hydrochloric acid adjust pH to 6 then with the dilution of 150ml water, adds the phosphoric acid buffer dilution, and the EA extraction of using 4 * 60ml again is to remove the sulfanilamide (SN) by product in the organic phase; Water is used hydrochloric acid adjust pH to 2 again, extracts with 3 * 60ml EA; Merge organic phase, with sal epsom (MgSO
4) drying, drying under reduced pressure obtains flavous oily liquids, is Azido-glycine.
Can be in the following ways to the purifying of final product integrin ligand cyclic peptide:
High performance liquid preparative chromatography: the C18 post, A is the 1 ‰ TFA/ aqueous solution mutually, the B phase: acetonitrile, thick peptide linearity and cyclisation are 5%-35%B, the 20min gradient, gradient is determined according to analyzing the chromatogram appearance time.
Synthetic route of the present invention is:
At first glycine is at trifluoromethanesulfanhydride anhydride, the effect of sodium azide generates Azido-glycine down, Fmoc-Asp (tBu)-OH generates Fmoc-Asp-Propargyl with the propargylamine condensation in solution, condensation Fmoc-Gly-OH successively after hanging over the side chain carboxyl group of alkynes propionyl aspartic acid on the 2-CTC resin then, Fmoc-Arg (Pbf)-OH, Fmoc-Xaa-OH and Azido-glycine generate linear tetrapeptide analogs, tetrapeptide analogs with linearity scales off from resin again, (DCM makes solvent under the condition of liquid phase, CuBr/DBU makes catalyzer), 4-6 hour synthetic cyclic peptide analogue of reaction under the room temperature: ring [arginine-glycine-aspartic acid-triazole ring generation-glycine-variable amino acid-] is Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Xaa-].
Cyclic peptide analogue cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Xaa-] synthetic route (Scheme 1.) be
Scheme?1.Strategy?for?synthesis?of?cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Xaa-].Reagents?and?conditions：(i)(1)DCC/HOBt，THF，pH＝7，r.t.，2h；(2)TFA/DCM＝2∶3(V∶V)；(ii)TBTU，NMM，20℃～40℃，2h～4h；(iii)(1)20％piperidine/DMF，30～60min；(2)EDC/HOBt，DIPEA，r.t.，4h～6h；(iv)CuBr/DBU＝1∶3，DCM，r.t.，6h；(2)TFA/H2O/TIPS＝95∶2.5∶2.5.
1. the synthetic route of nitrine glycine (Azido-glycine) (Scheme 2.) is
Scheme?2.Synthesis?of?Azido-glycine.
2. the synthetic route of alkynes propionyl aspartic acid (Fmoc-Asp-Propargyl) (Scheme 3.) is
Scheme?3.Synthesis?of?Fmoc-Asp-Propargyl
3. cyclisation route (Scheme 4.) is
Scheme?4.Synthesis?for?a?series?of?cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Xaa-]
Compared with prior art, the invention has the beneficial effects as follows:
(1) integrin ligand cyclic peptide analogues among the present invention has been introduced nitrogen heterocyclic (triazole ring), may increase biological activity;
(2) cyclization method of integrin ligand cyclic peptide analogues of the present invention is easier, efficient, mild condition.
Embodiment
The present invention is described in detail below by specific embodiment.
1. synthetic (principle that shifts according to diazonium) of nitrine glycine (Azido-glycine):
1.1 the preparation of fluoroform sulfonyl azide:
Sodium azide (6g) is dissolved in the round-bottomed flask of 150ml, adds the DCM of 24ml again, ice bath is cooled to 0 ℃, stirs 20 minutes.Reaction mixture with DCM extraction (2 * 12ml). merge organic phase,, promptly obtain being dissolved in the fluoroform sulfonyl azide of DCM, need not to be further purified next step reaction of direct adding and use with the saturated sodium carbonate washing.
1.2 the preparation of nitrine glycine:
With glycine (0.675g), K
2CO
3(1.783g), CuSO
4(0.0226g) join in the round-bottomed flask, add entry (30ml) again, methyl alcohol (60ml) and contain the DCM (48ml) of fluoroform sulfonyl azide at room temperature stirs, and reaction is spent the night.Subsequently, its decompression rotary evaporation is removed organic solvent, remaining part water (150ml) dilution. use hydrochloric acid adjust pH to 6 then, add the phosphoric acid buffer dilution, (4 * 60ml) extractions are to remove the sulfanilamide (SN) by product in the organic phase to use EA again.Water is used hydrochloric acid adjust pH to 2 again, with EA extraction (3 * 60ml). merge organic phase, use MgSO
4Drying, drying under reduced pressure obtains flavous oily liquids.
2. the preparation of alkynes propionyl aspartic acid:
Fmoc-Asp (OtBu)-OH (0.93g) is dissolved in the round-bottomed flask of THF (25ml) adding 100ml, add DCC then successively, (2.49g) and HOBt (1.62g). propargylamine adds in the reactor after with THF (5ml) mixing, reaction 2h, with triketohydrindene hydrate indicator detection reaction, with EA (3 * 20ml) and water (10ml) wash.With TFA/DCM (V: V=2: 3) remove side chain carboxyl group blocking group OtBu, with EA (3 * 20ml) and water (10ml) wash.Rotary evaporation steams solvent, and vacuum-drying obtains white solid, i.e. Fmoc-Asp-propargyl.
3. Fmoc-Asp-propargyl is hung on the 2-CTC resin tetrapeptide analogs of synthesizing linear:
Take by weighing 1g 2-CTC resin in solid phase reactor, add DCM 15ml swelling, washing resin.Add Fmoc-Asp-propargyl and DIPEA in the good resin of swelling, reaction is 2.5 hours under the room temperature, with EA (3 * 20ml) and water (10ml) wash.Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes.With EA (3 * 20ml) and water (10ml) washing, add Fmoc-Gly-OH, HBTU and NMM to reactor, react 2 hours, with triketohydrindene hydrate indicator detection reaction, with EA (3 * 20ml) and water (10ml) wash.Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes.With EA (3 * 20ml) and water (10ml) washing.Adopt the identical operations step, condensation Fmoc-Arg (Pbf)-OH, Fmoc-Xaa-OH and Azido-glycine generate linear tetrapeptide analogs successively.
4. cyclisation: the tetrapeptide analogs of the full guard of linearity is scaled off from resin, with lysate TFA/DCM (2: 8), can add several AcOH, cracking 2 hours is spin-dried for lysate.(DCM makes solvent under the condition of liquid phase; CuBr/DBU makes catalyzer); reaction is 4-6 hour under the room temperature; revolve DCM after the reaction, add lysate and remove side chain protected cracking 2h, revolve lysate; ether sedimentation on the rocks; the centrifugal supernatant liquor of removing obtains solid, i.e. cyclic peptide analogue Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Xaa-], send the analysis purifying.
5. purification condition
Analyze: high-efficient liquid phase analysis chromatogram (RP-HPLC), the C18 post, moving phase: A is the 1 ‰ TFA/ aqueous solution mutually.The B phase: acetonitrile, thick peptide linearity and cyclisation are 5%-35%B, the 20min gradient.The ultraviolet detection wavelength is 210nm.
Purifying: high performance liquid preparative chromatography: the C18 post, A is all the same with analysis condition mutually with B mutually.
Embodiment 1:
1.Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Gly-] (7a) synthetic:
1. synthetic (principle that shifts according to diazonium) of nitrine glycine (Azido-glycine):
1.1 the preparation of fluoroform sulfonyl azide:
Sodium azide (6g) is dissolved in the round-bottomed flask of 150ml, adds the DCM of 24ml again, ice bath is cooled to 0 ℃, stirs 20 minutes.Reaction mixture with DCM extraction (2 * 12ml). merge organic phase,, promptly obtain being dissolved in the fluoroform sulfonyl azide of methylene dichloride, need not to be further purified next step reaction of direct adding and use with the saturated sodium carbonate washing.
1.2 the preparation of nitrine glycine:
With glycine (0.675g), K
2CO
3(1.783g), CuSO
4(0.0226g) join in the round-bottomed flask, add entry (30ml) again, methyl alcohol (60ml) and contain the methylene dichloride (48ml) of fluoroform sulfonyl azide at room temperature stirs, and reaction is spent the night.Subsequently, its decompression rotary evaporation is removed organic solvent, remaining part water (150ml) dilution. use hydrochloric acid adjust pH to 6 then, add the phosphoric acid buffer dilution, (4 * 60ml) extractions are to remove the sulfanilamide (SN) by product in the organic phase to use EA again.Water is used hydrochloric acid adjust pH to 2 again, with EA extraction (3 * 60ml). merge organic phase, use MgSO
4Drying, drying under reduced pressure obtains flavous oily liquids.
2. the preparation of alkynes propionyl aspartic acid:
Fmoc-Asp (OtBu)-OH (0.93g) is dissolved in the round-bottomed flask of THF (25ml) adding 100ml, adding DCC (2.49g) and HOBt (1.62g) then successively. propargylamine adds in the reactor after using THF (5ml) mixing, reaction 2h, with triketohydrindene hydrate indicator detection reaction, with EA (3 * 20ml) and water (10ml) wash.With TFA/DCM (V: V=2: 3) remove side chain carboxyl group blocking group OtBu., with EA (3 * 20ml) and water (10ml) wash.Rotary evaporation steams solvent, and vacuum-drying obtains white solid, i.e. Fmoc-Asp-propargyl.
3. Fmoc-Asp-propargyl is hung on the 2-CTC resin tetrapeptide analogs of synthesizing linear:
Take by weighing 1g 2-CTC resin in solid phase reactor, add DCM 15ml swelling, washing resin.Added under Fmoc-Asp-propargyl and the DIPEA room temperature reaction in the good resin of swelling 2.5 hours, with EA (3 * 20ml) and water (10ml) wash.Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes.With EA (3 * 20ml) and water (10ml) washing, add Fmoc-Gly-OH, HBTU and NMM to reactor, react 2 hours, with triketohydrindene hydrate indicator detection reaction, with EA (3 * 20ml) and water (10ml) wash.Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes.With EA (3 * 20ml) and water (10ml) washing.Adopt the identical operations step, condensation Fmoc-Arg (Pbf)-OH, Fmoc-Gly-OH and Azido-glycine generate linear tetrapeptide analogs successively.
4. cyclisation: the tetrapeptide analogs of the full guard of linearity is scaled off from resin, with lysate TFE/DCM (2: 8), can add several AcOH, cracking 2 hours is spin-dried for lysate.(DCM makes solvent under the condition of liquid phase; CuBr/DBU makes catalyzer); reaction is 4-6 hour under the room temperature; revolve DCM after the reaction, add lysate and remove side chain protected cracking 2 hours, revolve lysate; ether sedimentation on the rocks; the centrifugal supernatant liquor of removing obtains solid, i.e. cyclic peptide analogue Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Gly-], send the analysis purifying.
5. purification condition
Analyze: high-efficient liquid phase analysis chromatogram (RP-HPLC), the C18 post, moving phase: A is the 1 ‰ TFA/ aqueous solution mutually.The B phase: acetonitrile, thick peptide linearity and cyclisation are 5%-35%B, the 20min gradient.The ultraviolet detection wavelength is 210nm.
Purifying: high performance liquid preparative chromatography: the C18 post, A is all the same with analysis condition mutually with B mutually.
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Gly-](7a).White?solids，mp：162-164℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.40(2H，d)，1.50-1.56(1H，m)，1.71-1.79(1H，m)，2.71-2.77(2H，m)，3.07-3.11(2H，m)，3.48(1H，J＝5.0Hz，d)，3.69(1H，J＝5.8Hz，d)，3.79-3.89(2H，m)，4.26(2H，J＝14.2，6.9Hz，dd)，4.45(2H，s)，4.54(2H，J＝6.3Hz，d)，5.05(1H，s)，5.29(1H，s)，7.50(1H，s)，7.68(2H，s)，7.83(1H，J＝5.9Hz，d)，8.03(1H，J＝8.7Hz，t)，8.28(1H，J＝5.9Hz，d)，8.42(1H，J＝7.1Hz，t)，8.84(1H，J＝5.8Hz，t)；
13C?NMR(125MHz，DMSO-d
6)：δ26.2，26.6，29.6，35.3，35.9，36.5，42.9，43.3，44.3，51.5，52.8，124.4，157.8，167.8，169.7，170.0，171.7，172.5，172.8；IR(KBr，cm
-1)：3393，3073，2932，1667，1538，1417，1331，1201，1136，1058，1024；HRMS：Calcd?for?C
19H
30N
11O
7(M+H)
+，524.2341，Found，524.2369.
Embodiment 2:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Ala-] (7b) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-L-Ala (Fmoc-Ala-OH) and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Ala-](7b).White?solids，mp：164-165℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.23(1H，J＝6.3Hz，d)，1.28(2H，J＝7.0Hz，d)，1.38-1.45(2H，m)，1.87-1.93(2H，m)，2.67-2.75(2H，m)，2.89(2H，s)，4.12-4.19(3H，m)，4.21(1H，J＝5.0Hz，d)，4.28(1H，J＝6.3Hz，d)，4.39(1H，J＝14.3，7.2Hz，dd)，4.40(1H，J＝7.1Hz，d)，5.07(1H，J＝6.3Hz，d)，5.27(1H，J＝16.6Hz，d)，6.92(2H，s)，7.36(2H，s)，7.63(1H，s)，7.88(1H，J＝7.8Hz，d)，8.26(1H，J＝8.5Hz，d)，8.35(1H，J＝6.9Hz，t)，8.48(1H，J＝7.5Hz，d)，8.78(1H，J＝5.8Hz，d)，12.38(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ18.6，24.5，26.2，27.1，29.4，30.0，32.9，36.1，43.0，49.1，51.2，54.6，124.5，157.8，169.9，170.2，171.7，172.8，173.0，173.6；IR(KBr，cm
-1)：3361，3063，2980，2936，1673，1538，1426，1384，1326，1202，1133，1058；HRMS：Calcd?for?C
20H
32N
11O
7(M+H)
+，538.2481，Found，538.2468
Embodiment 3:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Leu-] (7c) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-leucine (Fmoc-Leu-OH) and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Leu-](7c).White?solids，mp：167-169℃；
(c＝1.0?in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ0.86(3H，J＝6.5Hz，d)，0.91(3H，J＝6.5Hz，d)，1.47-1.55(5H，m)，1.69-1.76(2H，m)，2.74-2.82(2H，m)，3.09(2H，J＝6.8Hz，t)，3.94(1H，J＝6.8Hz，d)，4.05(2H，s)，4.21(1H，s)，4.44(1H，J＝14.1，7.1Hz，dd)，4.65(1H，J＝16.3Hz，d)，5.03(1H，J＝6.3Hz，d)，5.28(1H，J＝6.3Hz，d)，6.93(2H，s)，7.37(2H，s)，7.61(1H，s)，7.86(1H，J＝7.5Hz，d)，8.09(1H，J＝6.0Hz，t)，8.35(1H，J＝6.6Hz，d)，8.47(1H，J＝7.7Hz，d)，8.63(1H，J＝6.6Hz，d)，12.22(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ22.1，23.7，24.9，25.7，30.1，35.7，36.3，42.6，50.7，52.6，52.7，53.8，123.7，147.0，157.4，167.0，169.4，171.2，172.3，172.5，172.8；IR(KBr，cm
-1)：3404，3084，2959，2871，1667，1538，1470，1417，1384，1202，1138，1057，1019；HRMS：Calcd?for?C
23H
38N
11O
7(M+H)
+，580.2939.Found，580.2975.
Embodiment 4:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Ile-] (7d) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-Isoleucine (Fmoc-Ile-OH) and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Ile-](7d).Yellow?solids，mp：167-168℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ0.82-0.91(6H，m)，1.40-1.47(4H，m)，1.62-1.74(2H，m)，1.80-1.87(1H，m)，2.74-2.81(2H，m)，3.09(2H，J＝6.3Hz，d)，3.95(1H，J＝6.4Hz，d)，4.02(2H，J＝7.0Hz，d)，4.10(1H，J＝5.3Hz，d)，4.44(1H，J＝14.2，7.5Hz，dd)，4.88(1H，s)，5.11(1H，s)，5.28(1H，J＝16.1Hz，d)，6.95(2H，s)，7.37(2H，s)，7.63(1H，s)，8.11(1H，J＝6.3Hz，d)，8.27(1H，J＝8.4Hz，t)，8.36(1H，J＝6.9Hz，d)，8.44(1H，J＝7.9Hz，d)，8.52(1H，J＝6.6Hz，d)，12.35(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ12.4，12.5，25.9，29.7，30.3，36.4，42.9，50.8，51.0，53.1，53.4，53.7，59.3，60.2，124.2，147.4，157.8，167.1，169.8，171.6，172.3，172.9，173.0；IR(KBr，cm
-1)：3404，3084，2959，2871，1667，1538，1470，1417，1384，1202，1138，1057，1019；HRMS：Calcd?for?C
23H
38N
11O
7(M+H)
+，580.2950.Found，580.2946.
Embodiment 5:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Cys-] (7e) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-trityl-halfcystine [Fmoc-Cys (Trt)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Cys-](7e).White?solids，mp：189-190℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.36(2H，J＝6.5Hz，d)，1.44-1.52(1H，m)，1.62-1.71(1H，m)，2.53-2.64(2H，m)，2.89(2H，J＝6.2Hz，d)，3.69(1H，J＝6.1Hz，d)，3.72(1H，J＝6.4Hz，d)，4.31(H，J＝6.6Hz，d)，4.39(2H，J＝14.3，7.1Hz，dd)，4.56(2H，J＝6.3Hz，d)，4.61(2H，J＝6.3Hz，d)，6.16(2H，J＝7.5Hz，d)，6.79(2H，s)，7.23(2H，s)，7.80(1H，s)，8.12(1H，J＝6.7Hz，d)，8.29(1H，J＝7.1Hz，t)，8.47(1H，J＝6.6Hz，d)，8.54(1H，J＝6.7Hz，t)，8.75(1H，J＝6.8Hz，t)，12.36(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ24.2，26.3，27.4，27.7，29.2，30.5，32.3，36.1，43.3，49.3，51.7，56.3，124.8，156.8，169.3，170.5，171.7，172.3，173.2，173.9；IR(KBr，cm
-1)：3373，3079，2926，2849，1667，1538，1417，1384，1331，1201，1136，1057，1019；HRMS：Calcd?for?C
23H
38N
11O
7S(M+H)
+，570.2183.Found，570.2142.
Embodiment 6:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Met-] (7f) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-methionine(Met) (Fmoc-Met-OH) and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Met-](7f).Yellow?solids，mp：193-195℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.42-1.49(2H，m)，1.60-1.67(1H，m)，1.76-1.84(1H，m)，1.97(1H，s)，2.02(1H，s)，1.92-2.05(3H，m)，2.34-2.42(2H，m)，2.67(1H，s)，3.09(3H，J＝6.0Hz，d)，3.72-3.83(2H，m)，4.28(2H，J＝15.3，6.7Hz，dd)，4.41(1H，s)，4.57(1H，J＝5.4Hz，d)，5.17(2H，J＝14.8Hz，d)，7.49(1H，s)，7.79(1H，J＝8.9Hz，d)，8.21-8.27(1H，m)，8.33(1H，s)，8.42(1H，s)，8.60(1H，s)，12.38(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ15.8，26.229.4，30.6，33.5，35.7，37.5，41.2，41.6，43.1，50.7，53.3，53.5，125.5，129.6，145.7，157.9，166.6，169.8，171.4，171.9，172.8，172.9；IR(KBr，cm
-1)：3383，3073，2932，2849，1661，1538，1434，1384，1201，1137，1047；HRMS：Calcd?for?C
22H
36N
11O
7S(M+H)
+，598.2435.Found，598.2471.
Embodiment 7:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Asp-] (7g) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, the fluorenylmethyloxycarbonyl-aspartic acid-4-tert-butyl ester [Fmoc-Asp (OtBu)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Asp-](7g).White?solids，mp：193-195℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.41-1.44(2H，m)，1.56(2H，J＝9.4Hz，d)，2.57-2.63(1H，m)，2.76-2.83(1H，m)，3.07-3.11(2H，m)，3.90(2H，J＝6.3Hz，d)，4.13(1H，s)，4.25(1H，J＝4.7Hz，d)，4.40(1H，J＝14.7，7.7Hz，dd)，4.56-4.51(1H，m)，5.11(1H，J＝16.7Hz，d)，5.26(1H，J＝16.6Hz，d)，7.01(2H，s)，7.26(1H，s)，7.51(1H，s)，7.65(1H，s)，7.89(1H，J＝8.0Hz，d)，7.99(1H，J＝5.6Hz，t)，8.35(1H，J＝6.7Hz，d)，8.47(1H，J＝7.4Hz，d)，8.86(1H，J＝6.6Hz，d)，12.38(2H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ26.0，30.3，36.1，36.6，36.9，40.2，43.0，51.3，52.4，53.0，53.1，124.4，147.3，157.8，167.6，169.7，171.6，172.5，172.6，172.9；IR(KBr，cm
-1)：3375，3216，3073，2936，1667，1538，1416，1384，1331，1200，1137，1058，1024；HRMS：Calcd?for?C
21H
32N
11O
9(M+H)
+，582.5467.Found，582.5423.
Embodiment 8:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Asn-] (7h) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-trityl-l-asparagine [Fmoc-Asn (Trt)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Asn-](7h).White?solids，mp：187-190℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.43(2H，J＝6.8Hz，d)，1.52-1.59(1H，m)，1.78-1.84(1H，m)，2.76-2.87(2H，m)，3.01-3.09(4H，m)，3.53-3.58(2H，m)，3.89(1H，s)，4.13(1H，s)，4.38(1H，J＝14..4，7.1Hz，dd)，4.56(1H，s)，5.13(1H，J＝16.6Hz，d)，5.25(1H，J＝16.7Hz，d)，7.01(2H，s)，7.45(2H，s)，7.64(1H，s)，7.82(1H，s)，7.97(1H，s)，8.33(1H，J＝5.6Hz，t)，8.46(1H，J＝7.4Hz，d)，8.71(1H，J＝6.8Hz，d)，12.31(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ25.9，30.2，36.0，36.5，37.4，41.5，42.9，51.3，52.4，52.9，53.1，124.4，147.2，157.8，167.5，169.7，171.6，172.0，172.2，172.6，172.9；IR(KBr，cm
-1)：3350，3210，3073，2931，1669，1538，1417，1386，1336，1201，1136，1059，1030；HRMS：Calcd?forC
21H
33N
12O
8(M+H)
+，581.2568.Found，581.2562.
Embodiment 9:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Glu-] (7i) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, the fluorenylmethyloxycarbonyl-L-glutamic acid-4-tert-butyl ester [Fmoc-Glu (OtBu)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Glu-](7i).Yellow?solids，mp：197-199℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.42(1H，s)，1.54(1H，s)，1.80(1H，s)，2.01(1H，s)，2.34-2.39(1H，m)，2.79(1H，J＝6.4Hz，t)，3.07-3.13(2H，m)，3.45-3.53(1H，m)，3.95(1H，J＝6.7Hz，d)，4.02-4.09(1H，m)，4.13(1H，J＝14.3，6.6Hz，dd)，4.24(1H，J＝5.0Hz，d)，4.41(1H，J＝7.1Hz，d)，4.58(1H，J＝6.3Hz，d)，5.07(1H，s)，5.33(1H，J＝16.5Hz，d)，7.57(2H，s)，7.87(1H，s)，8.01(1H，J＝5.5Hz，d)，8.34(1H，J＝5.7Hz，t)，8.46(1H，J＝7.4Hz，d)，8.71(1H，J＝6.2Hz，d)，12.36(2H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ26.1，27.6，30.5，31.4，36.1，36.6，42.9，51.2，52.9，53.1，55.3，124.3，147.3，157.9，159.3，159.5，167.7，169.8，171.6，172.7，172.9，174.8；IR(KBr，cm
-1)：3419，3073，2948，2558，1668，1652，1557，1538，1413，1380，1178，1052，1024；HRMS：Calcd?for?C
22H
34N
11O
9(M+H)
+，596.2546.Found，596.2544.
Embodiment 10:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Lys-] (7j) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-N ε-tertbutyloxycarbonyl-Methionin [Fmoc-Lys (Boc)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Lys-](7j).Whitc?solids，mp：178-180℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.42-1.49(4H，m)，1.59(4H，J＝4.7Hz，d)，2.50-2.53(2H，m)，2.78-2.85(4H，m)，3.13(2H，J＝6.2Hz，d)，4.08(1H，J＝6.2Hz，d)，4.12(1H，s)，4.26(1H，s)，4.47(2H，J＝14.2，7.0Hz，dd)，5.12(2H，J＝16.3Hz，d)，5.21(2H，s)，6.96(2H，s)，7.35(2H，s)，7.79(1H，s)，8.08(1H，J＝5.4Hz，t)，8.22(1H，J＝8.4Hz，d)，8.32(1H，s)，8.44(1II，J＝7.6Hz，t)，8.78(1H，J＝6.7Hz，t)，12.37(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ22.3，24.6，26.4，27.3，28.9，29.4，30.6，32.5，32.9，36.4，43.0，43.8，49.4，51.3，54.5，124.3，157.3，169.8，170.4，171.5，172.3，173.2，173.5；IR(KBr，cm
-1)：3393，3073，2932，1667，1538，1417，1338，1201，1136，1058，1024；HRMS：Calcd?forC
23H
39N
12O
7(M+H)
+，595.3056，Found，595.3085
Embodiment 11:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Arg-] (7k) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, Fmoc-Arg (Pbf)-OH and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Arg-](7k).White?solids，mp：162-164℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.41-1.50(2H，m)，1.55-1.63(4H，m)，1.71-1.80(2H，m)，2.74-2.83(1H，m)，3.12(4H，J＝5.8Hz，d)，3.45-3.52(1H，m)，3.94-4.00(2H，m)，4.14(1H，J＝4.7Hz，d)，4.23(1H，J＝5.5Hz，d)，4.43(1H，J＝14.4，7.0Hz，dd)，5.06(1H，J＝16.5Hz，d)，5.31(1H，s)，7.63(1H，s)，7.70(2H，s)，7.91(1H，J＝7.5Hz，d)，8.05(1H，J＝5.6Hz，t)，8.36(1H，J＝6.0Hz，t)，8.49(1H，J＝7.5Hz，d)，8.76(1H，J＝6.4Hz，d)，12.36(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ24.6，27.7，27.9，29.5，31.0，32.3，37.8，43.8，52.7，52.3，54.8，124.8，157.9，169.9，170.2，171.2，172.5，173.3，173.9；IR(KBr，cm
-1)：3393，3212，3073，2932，1667，1538，1417，1331，1201，1136，1058，1024；HRMS：Calcd?forC
23H
39N
14O
7(M+H)
+，623.3121，Found，623.3120.
Embodiment 12:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-His-] (7l) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-trityl-Histidine [Fmoc-Hi s (Trt)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-His-](7l).White?solids，mp：176-178℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.41(2H，J＝5.9Hz，d)，1.51-1.58(1H，m)，1.79(1H，s)，2.76(1H，J＝5.8Hz，d)，3.09-3.16(2H，m)，3.56(1H，J＝4.9Hz，d)，3.90-3.96(1H，m)，4.16(1H，J＝5.3Hz，d)，4.25(1H，J＝5.3Hz，d)，4.41(2H，J＝7.4Hz，d)，4.53(1H，J＝14.2，7.0Hz，dd)，5.06(1H，J＝16.6Hz，d)，5.29(2H，J＝16.4Hz，d)，7.45(1H，s)，7.61(2H，J＝6.9Hz，t)，7.94(1H，J＝7.5Hz，d)，8.08(1H，J＝5.8Hz，t)，8.36(1H，J＝6.0Hz，d)，8.47(1H，J＝7.2Hz，d)，8.85(1H，J＝7.2Hz，d)，8.99(1H，J＝6.3Hz，d)，12.40(1H，s)；
13CNMR(125MHz，DMSO-d
6)：δ26.0，27.4，30.4，36.1，36.7，41.4，43.0，51.3，53.0，53.2，54.8，118.3，124.3，135.2，147.3，157.8，167.6，169.7，171.0，171.7，172.5，172.8；IR(KBr，cm
-1)：3393，3073，2932，1667，1538，1417，1331，1201，1136，1058，1024；HRMS：Calcd?for?C
23H
34N
13O
7(M+H)
+，604.2653，Found，604.2672.
Embodiment 13:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Phe-] (7m) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-phenylalanine [Fmoc-Phe-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Phe-](7m).Yellow?solids，mp：187-189℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.52-1.61(2H，m)，1.88-1.97(2H，m)，2.63-2.75(2H，m)，2.89(1H，s)，3.11(3H，J＝5.9Hz，d)，3.82(1H，J＝14.3，6.3Hz，dd)，4.19(1H，J＝14.2，6.5Hz，dd)，4.23(1H，J＝5.8Hz，d)，4.30(1H，J＝7.1Hz，d)，4.58(1H，s)，5.01(1H，s)，5.04(1H，s)，5.16(1H，s)，7.18-7.27(5H，m)，7.61(1H，J＝7.0Hz，d)，8.15(1H，s)，8.34(1H，J＝6.0Hz，t)，8.46(1H，J＝7.6Hz，d)，8.96(1H，s)，12.35(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ26.1，28.5，29.4，30.4，36.1，36.7，37.8，50.5，50.6，51.1，52.6，53.0，53.2，55.2，124.2，127.6，129.4，129.7，130.3，130.4，138.7，167.4，169.8，171.6，172.7，173.5；IR(KBr，cm
-1)：3352，3068，2931，2849，1667，1538，1457，1434，1384，1331，1201，1136，1084，1033；HRMS：Calcd?for?C
26H
36N
11O
7(M+H)
+，614.2794，Found，614.2758.
Embodiment 14:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Trp-] (7n) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-tertbutyloxycarbonyl-tryptophane [Fmoc-Trp (Boc)-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Trp-](7n).Yellow?solids，mp：173-175℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO?d
6)：δ1.38(1H，s)，1.51-1.58(2H，m)，1.77(1H，s)，3.04-3.10(2H，m)，3.16(1H，s)，3.44(1H，s)，3.50-3.57(1H，m)，3.65(1H，J＝5.2Hz，d)，4.23(1H，J＝5.7Hz，d)，4.35(1H，s)，4.43(2H，J＝14.2，6.6Hz，dd)，4.72(1H，s)，5.05(1H，s)，5.17(1H，s)，6.99-7.10(2H，m)，7.33-7.37(2H，m)，7.507.56(2H，m)，7.60(1H，s)，7.93(1H，s)，8.10(1H，s)，8.33(1H，J＝9.4Hz，d)，8.48(1H，J＝7.5Hz，t)，8.67(1H，s)，10.91(1H，J＝12.6Hz，d)，12.37(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ26.0，28.2，30.0，36.7，42.9，51.1，53.0，53.2，56.7，110.8，112.6，119.2，119.6，122.2，124.2，124.8，28.2，137.3，147.3，157.8，167.5，169.8，171.7，172.1，172.6，172.7；IR(KBr，cm
-1)：3393，3073，2932，1667，1538，1417，1331，1201，1136，1058，1024；HRMS：Calcd?for?C
28H
37N
12O
7(M+H)
+，653.2846，Found，653.2812.
Embodiment 15:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Pro-] (7o) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, fluorenylmethyloxycarbonyl-proline(Pro) [Fmoc-Pro-OH] and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Pro-](7o).White?solids，mp：162-164℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.34-1.41(4H，m)，1.65-1.74(6H，m)，2.53-2.64(4H，m)，2.89(2H，s)，3.68(1H，J＝7.2Hz，d)，4.10(1H，J＝6.6Hz，d)，4.27(1H，s)，4.77(2H，J＝14.2，6.7Hz，dd)，5.31(2H，J＝17.3Hz，d)，5.58(2H，J＝17.3Hz，d)，7.30(1H，s)，7.35(1H，J＝7.7Hz，t)，7.74(1H，s)，8.15(1H，J＝8.7Hz，d)，8.38(1H，J＝4.7Hz，t)，8.69(1H，J＝7.2Hz，d)，12.39(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ24.2，25.7，27.3，27.9，29.4，30.6，31.4，32.7，37.9，43.9，47.4，49.3，52.6，64.8，124.4，157.3，169.7，170.5，171.8，172.9，173.5，173.8；IR(KBr，cm
-1)：3385，2946，2746，1629，1585，1451，1376，1317，1267，1170，1037；HRMS：Calcd?for?C
22H
34N
11O
7(M+H)
+，564.2637，Found，564.2622.
Embodiment 16:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-] (7p) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-](7p).White?solids，mp：153-155℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.51-1.63(2H，m)，1.73-1.85(2H，m)，2.69-2.79(2H，m)，3.11(2H，J＝4.5Hz，d)，3.72(2H，s)，3.87(1H，s)，4.35(2H，J＝14.2，6.9Hz，dd)，4.50(1H，s)，4.60(1H，s)，5.15(1H，s)，7.21(1H，s)，7.54(2H，J＝5.1Hz，s)，7.81(1H，s)，8.24(1H，J＝7.8Hz，d)，8.37(1H，s)，8.65(1H，J＝4.9Hz，d)，12.39(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ24.8，27.6，28.6，29.6，31.1，32.4，37.9，49.8，52.4，54.7，124.2，157.6，169.1，171.3，172.4，173.5，173.8；IR(KBr，cm
-1)：3393，3073，2932，1667，1538，1417，1331，1201，1136，1058，1024；HRMS：Calcd?for?C
17H
27N
10O
6(M+H)
+，567.2039，Found，567.2056.
Embodiment 17:
Cyclo[-Arg-Gly-Asp-ψ (triazole)-Gly-Gly-Gly-] (7q) synthetic:
Step in the present embodiment is identical with embodiment 1, wherein:
In the 3rd step, condensation Fmoc-Arg (Pbf)-OH, Fmoc-Gly-OH, Fmoc-Gly-OH and Azido-glycine generate linear tetrapeptide analogs successively.
The physico-chemical analysis data of gained final product are as follows:
Cyclo[-Arg-Gly-Asp-ψ(triazole)-Gly-Gly-Gly-](7q).White?solids，mp：201-203℃；
(c＝1.0in?CHCl
3)；
1H?NMR(500MHz，DMSO-d
6)：δ1.10(2H，J＝9.5Hz，d)，1.31-1.42(1H，m)，1.60-1.71(1H，m)，2.56-2.62(2H，m)，2.76(2H，J＝4.8Hz，d)，3.69(1H，J＝5.1Hz，d)，3.75(1H，s)，3.86(2H，J＝6.8Hz，d)，4.35(3H，J＝14.2，6.7Hz，dd)，4.46(2H，J＝4.9Hz，d)，5.22(3H，J＝10.9Hz，d)，6.72(2H，s)，7.34(2H，J＝8.4Hz，d)，7.84(1H，s)，8.07(1H，s)，8.35(1H，J＝7.6Hz，t)，8.41(1H，J＝5.7Hz，t)，8.55(1H，J＝6.0Hz，t)，9.09(1H，J＝5.3Hz，t)，12.41(1H，s)；
13C?NMR(125MHz，DMSO-d
6)：δ25.9，29.1，36.1，36.7，41.5，43.6，44.2，44.9，51.3，52.4，52.7，125.1，146.3，157.7，168.7，170.2，170.5，171.6，173.2；IR(KBr，cm
-1)：3393，3073，2932，1667，1538，1417，1331，1201，1136，1058，1024；HRMS：Calcd?for?C
21H
33N
12O
8(M+H)
+，581.2597，Found，581.2569.
Claims (3)
1, a kind of integrin ligand cyclic peptide analogues, its structure is as follows:
2, the cyclization method of the described integrin ligand cyclic peptide analogues of a kind of claim 1 may further comprise the steps:
(1) Fmoc-Asp-propargyl is hung on the 2-CTC resin tetrapeptide analogs of synthesizing linear:
Take by weighing 1g 2-CTC resin in solid phase reactor, add the DCM15ml swelling, washing resin; In the good resin of swelling, add Fmoc-Asp-propargyl and DIPEA, react 2.5h under the room temperature, with the water washing of EA and the 10ml of 3 * 20ml; Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes; With the water washing of EA and the 10ml of 3 * 20ml, add Fmoc-Gly-OH, benzotriazole-N, N, N ', N '-tetramethyl-urea phosphofluoric acid ester and N-methylmorpholine react 2h to reactor, with triketohydrindene hydrate indicator detection reaction, with the water washing of EA and the 10ml of 3 * 20ml; Piperidines with 20%/DMF solution removes Fmoc, reacts 20 minutes; Water washing with EA and the 10ml of 3 * 20ml;
Adopt the identical operations step, condensation Fmoc-Arg (Pbf)-OH, Fmoc-Xaa-OH and Azido-glycine generate linear tetrapeptide analogs successively.
(2) cyclisation:
The tetrapeptide analogs of the full guard of linearity is scaled off from resin, and with 2: 8 TFE/DCM of lysate and several AcOH, cracking 2h is spin-dried for lysate; Make solvent at DCM, CuBr/DBU does to react 4～6 hours under the liquid-phase condition of catalyzer and the room temperature, revolves DCM after the reaction; add lysate and remove side chain protected cracking 2h, revolve lysate, ether sedimentation on the rocks; the centrifugal supernatant liquor of removing obtains solid, is the final product integrin ligand cyclic peptide.
3, the cyclization method of integrin ligand cyclic peptide analogues according to claim 2 is characterized in that, described Fmoc-Asp (OtBu)-OH makes in the following manner:
(1) the 6g sodium azide is dissolved in the round-bottomed flask of 150ml, adds the DCM of 24ml again, ice bath is cooled to 0 ℃, stirs 20 minutes; Reaction mixture extracts with 2 * 12mlDCM; Merge organic phase,, promptly obtain being dissolved in the fluoroform sulfonyl azide of DCM, need not to be further purified next step reaction of direct adding and use with the saturated sodium carbonate washing;
(2) with the K of 0.675g glycine, 1.783g
2CO
3, 0.0226g CuSO
4Join in the round-bottomed flask, add the DCM that 30ml water, 60ml methyl alcohol and 48ml contain the fluoroform sulfonyl azide again, at room temperature stir, reacted 24 hours; The rotary evaporation that subsequently reaction mixture reduced pressure is removed organic solvent, and remaining part is used hydrochloric acid adjust pH to 6 then with the dilution of 150ml water, adds the phosphoric acid buffer dilution, and the EA extraction of using 4 * 60ml again is to remove the sulfanilamide (SN) by product in the organic phase; Water is used hydrochloric acid adjust pH to 2 again, extracts with 3 * 60mlEA; Merge organic phase, use MgSO
4Drying, drying under reduced pressure obtains flavous oily liquids, is Fmoc-Asp (OtBu)-OH.
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| CN103588863B (en) * | 2013-11-15 | 2016-08-17 | 苏州强耀生物科技有限公司 | RGD cyclic peptide synthesis and preparation process |
| CN108137694A (en) * | 2015-09-18 | 2018-06-08 | 慕尼黑工业大学 | The ligand of αvβ6 Integrin and its synthesis and application |
| CN110590485A (en) * | 2018-06-13 | 2019-12-20 | 中国科学院上海有机化学研究所 | FSO2N3Use in the production of azide compounds |
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