CN117659119A - RGD fragment, method for preparing polypeptide by utilizing RGD fragment and application of RGD fragment - Google Patents
RGD fragment, method for preparing polypeptide by utilizing RGD fragment and application of RGD fragment Download PDFInfo
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- 229920001184 polypeptide Polymers 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 16
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical group NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 claims abstract description 71
- 238000005859 coupling reaction Methods 0.000 claims abstract description 58
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 claims abstract description 27
- 125000006239 protecting group Chemical group 0.000 claims abstract description 19
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 11
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 10
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- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 claims description 36
- 230000008878 coupling Effects 0.000 claims description 28
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- 150000001413 amino acids Chemical group 0.000 claims description 26
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 26
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- 238000003776 cleavage reaction Methods 0.000 claims description 16
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- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 15
- 238000009833 condensation Methods 0.000 claims description 14
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention discloses an RGD fragment, a method for preparing polypeptide by using the RGD fragment and application of the RGD fragment. The RGD fragment comprises Arg-Gly-Asp, pbf is modified on the side chain of Arg, otBu is modified on the side chain of Asp, and an amino protecting group is also arranged on Arg. The method provided by the invention has the advantages that the steps of coupling reaction required are reduced, the synthesis efficiency is improved, the side reaction is reduced, and the generation of corresponding missing impurities and inserted impurities is avoided.
Description
Technical Field
The invention belongs to the field of chemical synthesis, and particularly relates to an RGD fragment, a method for preparing polypeptide by using the RGD fragment and application of the RGD fragment.
Background
RGD peptides are a class of short peptides containing the sequence arginine-glycine-aspartic acid (Arg-Gly-Asp), which are widely found in organisms, and are recognition sites for integrins (integrins) to interact with their ligand proteins (Pierschbacher, M.D., ruoslahti, E.,1984.Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule.Nature309,30-33). Since the first report in 1984 that the RGD sequence contained in fibrinogen is a cell recognition site, RGD peptides and derivatives thereof have become a focus of attention and study by many scholars. The most common RGD sequence-containing matrix proteins in humans are mainly fibronectin, laminin, vitronectin, collagen, thrombospondin, and the like (Temming K, schiffemers R.M, molema G, et al Kok RJ.RGD-based strategies for selectivedelivery of therapeutics and imaging agents to the tumor vasculature [ J ]. Drug Resist Updat,2005,8 (6): 381-402).
RGD sequences are widely distributed in extracellular matrix proteins. Extracellular matrix proteins are identified and combined with integrin receptors on cell membranes through RGD sequences, mediate interactions between cells and extracellular matrixes and between cells, and participate in pathological processes such as angiogenesis, tumor generation, development, infiltration, metastasis and the like.
At present, when RGD peptide sequence exists in a peptide chain, amino acid coupling is mainly adopted one by one, so that the method has the advantages of complex steps, low efficiency and long time consumption, and is easy to introduce deletion impurities and insertion impurities.
Disclosure of Invention
In order to overcome the defects of complicated steps, low efficiency, long time consumption and more impurities, the invention provides an RGD fragment, a method for preparing polypeptide by using the RGD fragment and application of the RGD fragment; specifically, the invention adopts Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH amino acid solid phase to synthesize polypeptide containing RGD fragment.
To address the deficiencies of the prior art, a first aspect of the present invention provides a modified RGD fragment comprising Arg-Gly-Asp with Pbf modified on the side chain of Arg, otBu modified on the side chain of Asp, and an amine protecting group such as Fmoc (2-chlorobenzyloxycarbonyl) on Arg.
In certain embodiments, the RGD fragment is Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH.
The second aspect of the invention provides a synthesis method of RGD peptide, which comprises the steps of taking a solid phase material as a carrier, sequentially coupling amino acid residues modified by side chain protecting groups in a condensation system, finally coupling RGD fragments according to the first aspect of the invention, and removing the side chain protecting groups through cleavage to obtain the RGD peptide. Preferably, the RGD peptide is 30 peptide, and the amino acid sequence of the RGD peptide is shown as SEQ ID NO. 1. More preferably, the RGD fragments are coupled 2 times consecutively.
The third aspect of the invention provides a synthesis method of RGD peptide, which comprises the steps of taking a solid phase material as a carrier, coupling Fmoc-Gly-Gly-OH to the solid phase material in a condensation system, sequentially coupling amino acid residues modified by side chain protecting groups, and removing the side chain protecting groups through cleavage to obtain the RGD peptide. Preferably, the RGD peptide is 30 peptide, and the amino acid sequence of the RGD peptide is shown as SEQ ID NO. 1.
In certain embodiments, fmoc-Gly-Gly-OH is first attached to the solid phase material, amino acid residues modified by side chain protecting groups are coupled in sequence, then RGD fragments according to the first aspect of the invention are coupled 2 times, and the side chain protecting groups are removed by cleavage to obtain the RGD peptide.
In certain embodiments, the coupling reaction of the coupling proceeds from the C-terminus to the N-terminus, and/or the solid phase material is a resin; for example, the solid phase material is Rink amide AM Resin resin or Rink amide MHBA Resin resin. The resin may be a solid support resin ending in an amide at the C-terminus of the polypeptide obtained after cleavage.
In certain embodiments, the amino acid modified with a side chain protecting group is Fmoc-X-OH, wherein X is an amino acid residue, preferably the side chain of the amino acid residue is further modified with one or more of the following protecting groups: tBu, trt, otBu and Pbf.
In certain embodiments, the Fmoc-X-OH is Fmoc-Met-OH, fmoc-Ser (tBu) -OH, fmoc-Leu-OH, fmoc-Pro-OH, fmoc-Asn (Trt) -OH, fmoc-Ala-OH, fmoc-Val-OH, fmoc-His (Trt) -OH, fmoc-Gln (Trt) -OH, fmoc-Phe-OH, fmoc-Asp (OtBu) -OH, or Fmoc-Arg (Pbf) -OH.
In certain embodiments, the condensation system is HOBT/DIC, which is subjected to Fmoc deprotection prior to the coupling, preferably,
the Fmoc removal protection uses PIP/DMF solution;
the cleavage reagent used for the cleavage is TFA/Mpr/H 2 A mixed solution of O/DMS;
more preferably, the Fmoc removal protection uses a 20% PIP/DMF solution;
among the cleavage reagents, TFA/Mpr/H 2 The volume ratio of O/DMS is 92:2:2:2;
the RGD peptide is obtained by filtration, concentration and precipitation after cleavage, preferably by adding isopropyl ether for precipitation.
In certain embodiments, the coupling reaction of the coupling uses a condensing reagent of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDCI)/N-hydroxysuccinimide (HOSu), N-Carbodiimidazole (CDI)/base buffer, 1, 3-Dicyclohexylcarbodiimide (DCC)/N-Hydroxysuccinimide (HO)Su), trimethylacetyl chloride (PivCl) and thionyl chloride (SOCl) 2 ) One or more of Dichloromethane (DCM) is used for the coupling reaction at 10-40 deg.C and/or the time of 1-3.5 hr.
In certain embodiments, the coupling reaction temperature of the coupling is one of 15±2 ℃, 25±2 ℃ and 35±2 ℃; the coupling reaction time is one of 1h, 2h and 3 h.
In certain embodiments, the coupling reaction of the coupling comprises coupling the following protected amino acids sequentially from the C-terminus to the N-terminus: fmoc-Met-OH; fmoc-Gly-Gly-OH or Fmoc-Gly-OH, fmoc-Gly-OH; fmoc-Ser (tBu) -OH, fmoc-Leu-OH, fmoc-Pro-OH, fmoc-Ser (tBu) -OH, fmoc-Asn (Trt) -OH, fmoc-Leu-OH, fmoc-Ala-OH, fmoc-Val-OH, fmoc-Leu-OH, fmoc-His (Trt) -OH, fmoc-Leu-OH, fmoc-Val-OH, fmoc-Pro-OH, fmoc-Gln (Trt) -OH, fmoc-Phe-OH, fmoc-Asp (OtBu) -OH, fmoc-Arg (Pbf) -OH, fmoc-His (Trt) -OH, fmoc-Ser (tBu) -OH, fmoc-His (Trt) -OH, fmoc-Met-OH, fmoc-Gly (OtBu) -OH.
According to a fourth aspect of the present invention, there is provided a method for preparing an RGD fragment according to the first aspect of the present invention, the method comprising:
(a) Mixing and stirring amino acid Fmoc-Arg (Pbf) -OH and H-Gly-OH dissolution and condensation reagents to obtain Fmoc-Arg (Pbf) -Gly;
(b) Mixing Fmoc-Arg (Pbf) -Gly, H-Asp (OtBu) -OH and a condensation reagent, and stirring to obtain the RGD fragment;
the condensing agent in the (a) and the (b) is EDCI/HOSu, CDI/base, DCC/HOSu, pivCl and SOCl respectively 2 One or more of DCM;
preferably, the condensing agent is EDCI/HOSu.
In a fifth aspect, the present invention provides the use of an RGD fragment according to the first aspect of the present invention for the preparation of a polypeptide drug.
On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that:
1. the method for synthesizing Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH shortens the coupling reaction step in the process of polypeptide solid-phase synthesis of RGD sequence, and improves the synthesis efficiency.
2. The method for synthesizing the 30 peptide by using the Fmoc-Gly-Gly-OH amino acid fragment only needs 29 steps of coupling reaction, improves the synthesis efficiency, reduces side reaction and avoids the generation of corresponding missing impurities and inserted impurities.
3. The method for synthesizing the 30 peptide containing the RGD fragment by using the Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH amino acid fragment provided by the invention only needs 26 steps of coupling reaction, improves the synthesis efficiency, reduces side reactions and avoids the generation of corresponding missing impurities and inserted impurities.
4. The method for synthesizing the 30 peptide containing the RGD fragment by using the Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH and the Fmoc-Gly-Gly-OH amino acid fragment provided by the invention only needs 25 steps of coupling reaction, improves the synthesis efficiency, reduces side reactions and avoids the generation of corresponding missing impurities and inserted impurities.
Drawings
FIG. 1 shows the results of solid phase synthesis using Fmoc-Gly-Gly-OH.
FIG. 2 shows the results of solid phase synthesis using Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH.
FIG. 3 shows the results after amino acid-by-amino acid coupling.
FIG. 4 shows the results of solid phase synthesis using Fmoc-Gly-Gly-OH and Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
1. The specific route for synthesizing Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH provided by the invention is as follows:
2. the specific route for synthesizing the 30 peptide by adopting Fmoc-Gly-Gly-OH amino acid fragments is as follows:
3. a specific route for synthesizing a 30 peptide containing RGD fragment using Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH amino acid fragment is as follows:
4. a specific route for synthesizing a 30 peptide containing RGD fragment using Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH and Fmoc-Gly-Gly-OH amino acid fragments is as follows:
the sequence of the RGD peptide (30 peptide) after removing all protecting groups is as follows: RGDRGDMHSHRDFQPVLHLVALNSPLSGGM (SEQ ID NO: 1).
The invention is further described below in connection with specific embodiments.
Description of abbreviations appearing in the present invention:
fomc: 9-fluorenylmethoxycarbonyl
HOBt: 1-hydroxybenzotriazoles
DIC: n, N-diisopropylcarbodiimide
DMF: dimethylformamide
PIP: piperidine compounds
CDI: n, N-carbonyl diimidazoles
base: base buffer
DCC:1, 3-dicyclohexylcarbodiimide
HOSu: n-hydroxysuccinimide
PivCl: trimethyl acetyl chloride
SOCl 2 : thionyl chloride
DCM: dichloromethane (dichloromethane)
TFA: trifluoroacetic acid
Mpr: mercaptopropionic acid
DMS: dimethyl sulfide
CAS number for intermediate 1Fmoc-Arg (Pbf) -Gly: 660846-80-0
CAS number for H-Asp (OtBu) -OH: 154445-77-9
CAS number for Fmoc-Gly-Gly-OH: 35665-38-4
EXAMPLE 1 solid phase Synthesis of RGD peptide Using Fmoc-Gly-Gly-OH
1.1 preparation of Fmoc-Gly-Gly-OH
Fmoc-Gly-OH and H-Gly-OH, adding into a reaction kettle, and adding a proper amount of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDCI)/N-hydroxysuccinimide (HOSu); stirring for condensation reaction to obtain Fmoc-Gly-Gly-OH
1.2 Synthesis of polypeptide resins
The polypeptide resin is as follows: arg (Pbf) -Gly-Asp (OtBu) -Arg (Pbf) -Gly-Asp (OtBu) -Met-His (Trt) -Ser (tBu) -His (Trt) -Arg (Pbf) -Asp (OtBu) -Phe-Gln (Trt) -Pro-Val-Leu-His (Trt) -Leu-Val-Ala-Leu-Asn (Trt) -Ser (tBu) -Pro-Leu-Ser (tBu) -Gly-Gly-Met-Rink amide MHBA Resin resin.
The polypeptide resin was prepared by coupling with the protected amino acids shown in Table 1 in sequence by Fmoc deprotection and coupling reaction using Rink amide MHBA Resin resin as starting support. The protected amino acids corresponding to those used in this example are shown in Table 1 below, wherein Fmoc-Gly-Gly-OH fragments were used to reduce the number of coupling steps and improve the efficiency.
TABLE 1 protected amino acids
| Amino acid sequence number | Protecting amino acids | Molecular weight |
| 30 | Fmoc-Met-OH | 371.45 |
| 29-28 | Fmoc-Gly-Gly-OH | 354.36 |
| 27 | Fmoc-Ser(tBu)-OH | 383.44 |
| 26 | Fmoc-Leu-OH | 353.41 |
| 25 | Fmoc-Pro-OH | 337.37 |
| 24 | Fmoc-Ser(tBu)-OH | 383.44 |
| 23 | Fmoc-Asn(Trt)-OH | 596.67 |
| 22 | Fmoc-Leu-OH | 353.41 |
| 21 | Fmoc-Ala-OH | 311.33 |
| 20 | Fmoc-Val-OH | 339.39 |
| 19 | Fmoc-Leu-OH | 353.41 |
| 18 | Fmoc-His(Trt)-OH | 619.71 |
| 17 | Fmoc-Leu-OH | 353.41 |
| 16 | Fmoc-Val-OH | 339.39 |
| 15 | Fmoc-Pro-OH | 337.37 |
| 14 | Fmoc-Gln(Trt)-OH | 610.70 |
| 13 | Fmoc-Phe-OH | 387.43 |
| 12 | Fmoc-Asp(OtBu)-OH | 411.45 |
| 11 | Fmoc-Arg(Pbf)-OH | 648.77 |
| 10 | Fmoc-His(Trt)-OH | 619.71 |
| 9 | Fmoc-Ser(tBu)-OH | 383.44 |
| 8 | Fmoc-His(Trt)-OH | 619.71 |
| 7 | Fmoc-Met-OH | 371.45 |
| 6 | Fmoc-Asp(OtBu)-OH | 411.45 |
| 5 | Fmoc-Gly-Gly-OH | 354.36 |
| 4 | Fmoc-Arg(Pbf)-OH | 648.77 |
| 3 | Fmoc-Asp(OtBu)-OH | 411.45 |
| 2 | Fmoc-Gly-OH | 354.36 |
| 1 | Fmoc-Arg(Pbf)-OH | 648.77 |
The preparation is carried out from the C end to the N end according to the amino acid sequence number (from large to small). Rink amide MHBA Resin resin (substitution S=0.37 mmol/g) was taken. The condensation system adopts HOBT/DIC; the coupling reaction temperature is controlled at 25+/-5 ℃ and the coupling reaction time is 1.5-3.5h; deprotection (removal of Fmoc protection) with 20% PIP/DMF solution, e.g. (Pbf), (OtBu) is the side chain protecting group of the amino acid. S30-S16 deprotection time is 5+10min; the deprotection time of S15-S1 is 5+15min; filtering to obtain Fmoc-removed resin, namely polypeptide resin, for later use.
1.3 preparation of crude polypeptide
Taking the polypeptide resin prepared in example 1.2, adding TFA/Mpr/H in a volume ratio 2 The cleavage reagent, O/dms=92/2/2/2, was stirred well and reacted at room temperature for 4 hours, the reaction mixture was filtered using a sand funnel, filtrate 1 was collected, the resin was washed 3 times with a small amount of TFA (analytically pure reagent, AR) to collect filtrate 2, and filtrate 1 and filtrate 2 were combined. Concentrating under reduced pressure by rotary evaporation, mixing filtrates, adding concentrated solution into isopropyl ether, standing, precipitating, centrifuging, collecting solid precipitate, and vacuum drying to obtain white powder, which is the polypeptide crude product with purity of 62.93% shown in figure 1.
EXAMPLE 2 solid phase Synthesis of a 30 peptide containing RGDRGD fragment Using Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH
2.1 preparation of the starting materials according to the invention by liquid phase synthesis: fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH
Dissolving amino acids Fmoc-Arg (Pbf) -OH and H-Gly-OH, adding into a reaction kettle, and adding a proper amount of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDCI)/N-hydroxysuccinimide (HOSu); stirring for condensation reaction to obtain Fmoc-Arg (Pbf) -Gly (intermediate 1)
Adding dissolved H-Asp (OtBu) -OH to the intermediate 1, and adding a proper amount of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDCI)/N-hydroxysuccinimide (HOSu); the mixture was stirred for condensation reaction to give Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH.
2.2 preparation of polypeptide resin by Fmoc deprotection and coupling reaction with the protected amino acids shown in Table 2 in sequence using Rink amide MHBA Resin resin as starting support. The protected amino acids corresponding to those used in this example are shown in Table 2 below, wherein Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH fragments were used to reduce the number of coupling steps and increase the efficiency.
TABLE 2 protection of amino acids
The preparation is carried out from the C end to the N end according to the amino acid sequence number (from large to small). Rink amide MHBA Resin resin (substitution S=0.37 mmol/g) was taken. The condensation system adopts HOBT/DIC; the coupling reaction temperature is controlled at 25+/-5 ℃ and the coupling reaction time is 1.5-3.5h; deprotection with 20% PIP/DMF solution and filtration gave Fmoc-removed resin.
2.3 comparison of Effect of Fmoc-Gly-Gly-OH and Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH on the Synthesis of 30 peptide
The preparation is carried out from the C end to the N end according to the amino acid sequence number (from large to small). Rink amide MHBA Resin resin was taken. The condensation system adopts HOBT/DIC; the coupling reaction temperature is controlled at 25+/-5 ℃ and the coupling reaction time is 1.5-3.5h; deprotection with 20% PIP/DMF solution and filtration gave Fmoc-removed resin.
2.4 preparation of crude polypeptide the preparation method is the same as that of crude polypeptide 1.3 of example 1. Crude polypeptide with purity of 69.74% is shown in figure 2.
TABLE 3 comparison of different fragments versus purity and yield
| Fragments | Purity of crude product |
| Amino acid by amino acid coupling (FIG. 3) | 52.55％ |
| Fmoc-Gly-Gly-OH | 62.93％ |
| Fmoc-Arg(Pbf)-Gly-Asp(OtBu)-OH | 69.74％ |
The results of the table show that the target peptide is synthesized by adopting a fragment method, and the purity of the crude product is obviously improved. The peptide synthesized by Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH fragment reduces one-step synthesis and obviously improves the purity of the crude product compared with the peptide synthesized by Fmoc-Gly-Gly-OH fragment.
EXAMPLE 3 solid phase Synthesis of a 30 peptide containing GG and RGDRGD fragments Using Fmoc-Gly-Gly-OH and Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH
The polypeptide resin was prepared by coupling with the protected amino acids shown in Table 4 in sequence by Fmoc deprotection and coupling reaction using Rink amide MHBA Resin resin as a starting carrier. The protected amino acids corresponding to those used in this example are shown in Table 4 below, wherein Fmoc-Gly-Gly-OH and Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH fragments, reduced coupling reaction steps and improved efficiency.
TABLE 4 protected amino acids
The preparation is carried out from the C end to the N end according to the amino acid sequence number (from large to small). Rink amide MHBA Resin resin (substitution S=0.37 mmol/g) was taken. The condensation system adopts HOBT/DIC; the coupling reaction temperature is controlled at 25+/-5 ℃ and the coupling reaction time is 1.5-3.5h; deprotection with 20% PIP/DMF solution and filtration gave Fmoc-removed resin.
The preparation method of the crude polypeptide is the same as that of the 1.3 crude polypeptide in the embodiment 1. Crude polypeptide with purity of 74.07% is shown in figure 4.
TABLE 5 comparison of different fragments versus purity and yield
| Fragments | Purity of crude product |
| Amino acid by amino acid coupling (FIG. 3) | 52.55％ |
| Fmoc-Gly-Gly-OH | 62.93％ |
| Fmoc-Arg(Pbf)-Gly-Asp(OtBu)-OH | 69.74％ |
| Fmoc-Gly-Gly-OH and Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH | 74.07％ |
The results in Table 5 above demonstrate that RGD peptide synthesis using two amino acid fragments reduces and synthesizes steps, saves cost, and has the highest crude product purity, i.e., 74.07%.
Claims (14)
1. A modified RGD fragment comprising Arg-Gly-Asp, having Pbf modified in the side chain of Arg, otBu modified in the side chain of Asp, and further having an amine protecting group such as Fmoc (2-chlorobenzyloxycarbonyl) in Arg.
2. The RGD fragment of claim 1, which is Fmoc-Arg (Pbf) -Gly-Asp (OtBu) -OH.
3. The synthesis method of RGD peptide is characterized by comprising the steps of taking a solid phase material as a carrier, sequentially coupling amino acid residues modified by side chain protecting groups in a condensation system, finally coupling RGD fragments according to claim 1 or 2, and removing the side chain protecting groups through cleavage to obtain the RGD peptide; preferably, the RGD peptide is 30 peptide, and the amino acid sequence of the RGD peptide is shown as SEQ ID NO. 1; more preferably, the RGD fragments are coupled 2 times consecutively.
4. The synthesis method of RGD peptide is characterized by comprising the steps of taking a solid phase material as a carrier, coupling Fmoc-Gly-Gly-OH to the solid phase material in a condensation system, sequentially coupling amino acid residues modified by side chain protecting groups, and removing the side chain protecting groups through cleavage to obtain RGD peptide; preferably, the RGD peptide is 30 peptide, and the amino acid sequence of the RGD peptide is shown as SEQ ID NO. 1.
5. The synthesis method according to claim 3 or 4, wherein Fmoc-Gly-Gly-OH is first connected to the solid phase material, amino acid residues modified by side chain protecting groups are coupled in sequence, then RGD fragments according to claim 1 or 2 are coupled for 2 times, and the side chain protecting groups are removed through cleavage to obtain the RGD peptide.
6. The synthetic method according to any one of claims 3 to 5, wherein the coupling reaction of the coupling is performed from C-terminal to N-terminal, and/or the solid phase material is a resin; for example, the solid phase material is Rink amide AM Resin resin or Rink amide MHBA Resin resin.
7. The synthetic method according to any one of claims 3 to 6, wherein the amino acid modified with a side chain protecting group is Fmoc-X-OH, wherein X is an amino acid residue, preferably the side chain of the amino acid residue is further modified with one or more of the following protecting groups: tBu, trt, otBu and Pbf.
8. The synthetic method of claim 7, wherein the Fmoc-X-OH is Fmoc-Met-OH, fmoc-Ser (tBu) -OH, fmoc-Leu-OH, fmoc-Pro-OH, fmoc-Asn (Trt) -OH, fmoc-Ala-OH, fmoc-Val-OH, fmoc-His (Trt) -OH, fmoc-gin (Trt) -OH, fmoc-Phe-OH, fmoc-Asp (OtBu) -OH, or Fmoc-Arg (Pbf) -OH.
9. The synthesis according to any of claims 3 to 8, wherein the condensation system is HOBT/DIC, which is subjected to Fmoc protection prior to the coupling, preferably,
the Fmoc removal protection uses PIP/DMF solution;
the cleavage reagent used for the cleavage is TFA/Mpr/H 2 A mixed solution of O/DMS;
more preferably, the Fmoc removal protection uses a 20% PIP/DMF solution;
among the cleavage reagents, TFA/Mpr/H 2 The volume ratio of O/DMS is 92:2:2:2;
the RGD peptide is obtained by filtration, concentration and precipitation after cleavage, preferably by adding isopropyl ether for precipitation.
10. The synthesis method according to any one of claims 3 to 9, wherein the condensing reagent used in the coupling reaction of the coupling is EDCI/HOSu, CDI/bEnzyme, DCC/HOSu, pivCl and SOCl 2 One or more of DCM, the temperature of the coupling reaction is 10-40 ℃ and/or the time of the coupling reaction is 1-3.5 hours.
11. The synthetic method of claim 1 wherein the coupling reaction temperature of the coupling is one of 15±2 ℃, 25±2 ℃ and 35±2 ℃; the coupling reaction time is one of 1h, 2h and 3 h.
12. The synthetic method of any one of claims 5 to 11 wherein the coupling reaction of the coupling comprises coupling the following protected amino acids sequentially from C-terminus to N-terminus: fmoc-Met-OH; fmoc-Gly-Gly-OH or Fmoc-Gly-OH, fmoc-Gly-OH; fmoc-Ser (tBu) -OH, fmoc-Leu-OH, fmoc-Pro-OH, fmoc-Ser (tBu) -OH, fmoc-Asn (Trt) -OH, fmoc-Leu-OH, fmoc-Ala-OH, fmoc-Val-OH, fmoc-Leu-OH, fmoc-His (Trt) -OH, fmoc-Leu-OH, fmoc-Val-OH, fmoc-Pro-OH, fmoc-Gln (Trt) -OH, fmoc-Phe-OH, fmoc-Asp (OtBu) -OH, fmoc-Arg (Pbf) -OH, fmoc-His (Trt) -OH, fmoc-Ser (tBu) -OH, fmoc-His (Trt) -OH, fmoc-Met-OH, fmoc-Gly (OtBu) -OH.
13. A method of preparing an RGD fragment according to claim 1 or 2, comprising:
(a) Mixing and stirring amino acid Fmoc-Arg (Pbf) -OH and H-Gly-OH dissolution and condensation reagents to obtain Fmoc-Arg (Pbf) -Gly;
(b) Mixing Fmoc-Arg (Pbf) -Gly, H-Asp (OtBu) -OH and a condensation reagent, and stirring to obtain the RGD fragment;
the condensing agent in the (a) and the (b) is EDCI/HOSu, CDI/base, DCC/HOSu, pivCl and SOCl respectively 2 One or more of DCM;
preferably, the condensing agent is EDCI/HOSu.
14. Use of an RGD fragment according to claim 1 or 2 for the preparation of a polypeptide drug.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09278669A (en) * | 1996-04-17 | 1997-10-28 | Asahi Glass Co Ltd | Therapeutic agent for nephritis |
| US20050158301A1 (en) * | 2002-07-02 | 2005-07-21 | The Board Of Regents, The University Of Texas System | Thrombin peptide derivatives |
| US20060251699A1 (en) * | 2003-08-29 | 2006-11-09 | Children's Medical Center Corporation | Anti-angiogenic peptides from the N-terminus of endostatin |
| US20090257952A1 (en) * | 2006-10-04 | 2009-10-15 | The Board Of Trustees Of The Leland Stanford Junior University | Engineered Integrin Binding Peptides |
| KR20090132815A (en) * | 2008-06-23 | 2009-12-31 | (주)케어젠 | Peptides having activities of transforming growth factor-beta and their uses |
| CN101817873A (en) * | 2010-03-11 | 2010-09-01 | 复旦大学 | Cell adhesion promoting polypeptide and preparation method thereof |
| US20110160431A1 (en) * | 2009-04-06 | 2011-06-30 | Novetide, Ltd. | Production of peptides containing poly-gly sequences using fmoc chemistry |
| CN102924578A (en) * | 2011-08-09 | 2013-02-13 | 哈药集团技术中心 | Anti-tumor polypeptide, preparation method and anti-tumor applications thereof |
| CN104530199A (en) * | 2014-11-18 | 2015-04-22 | 哈尔滨医科大学 | Antitumor polypeptide, and preparation method and application thereof |
| CN104530224A (en) * | 2015-01-07 | 2015-04-22 | 哈尔滨吉象隆生物技术有限公司 | Bivalirudin solid-phase synthesis method |
| CN106146614A (en) * | 2015-05-14 | 2016-11-23 | 沈阳科海医药科技有限公司 | A kind of preparation method of RGD tripeptides |
-
2023
- 2023-10-31 CN CN202311430717.5A patent/CN117659119A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09278669A (en) * | 1996-04-17 | 1997-10-28 | Asahi Glass Co Ltd | Therapeutic agent for nephritis |
| US20050158301A1 (en) * | 2002-07-02 | 2005-07-21 | The Board Of Regents, The University Of Texas System | Thrombin peptide derivatives |
| US20060251699A1 (en) * | 2003-08-29 | 2006-11-09 | Children's Medical Center Corporation | Anti-angiogenic peptides from the N-terminus of endostatin |
| US20090257952A1 (en) * | 2006-10-04 | 2009-10-15 | The Board Of Trustees Of The Leland Stanford Junior University | Engineered Integrin Binding Peptides |
| KR20090132815A (en) * | 2008-06-23 | 2009-12-31 | (주)케어젠 | Peptides having activities of transforming growth factor-beta and their uses |
| US20110160431A1 (en) * | 2009-04-06 | 2011-06-30 | Novetide, Ltd. | Production of peptides containing poly-gly sequences using fmoc chemistry |
| CN101817873A (en) * | 2010-03-11 | 2010-09-01 | 复旦大学 | Cell adhesion promoting polypeptide and preparation method thereof |
| CN102924578A (en) * | 2011-08-09 | 2013-02-13 | 哈药集团技术中心 | Anti-tumor polypeptide, preparation method and anti-tumor applications thereof |
| CN104530199A (en) * | 2014-11-18 | 2015-04-22 | 哈尔滨医科大学 | Antitumor polypeptide, and preparation method and application thereof |
| CN104530224A (en) * | 2015-01-07 | 2015-04-22 | 哈尔滨吉象隆生物技术有限公司 | Bivalirudin solid-phase synthesis method |
| CN106146614A (en) * | 2015-05-14 | 2016-11-23 | 沈阳科海医药科技有限公司 | A kind of preparation method of RGD tripeptides |
Non-Patent Citations (4)
| Title |
|---|
| GENBANK: "AXE43015.1: recombinant human endostatin [synthetic construct]", 《GENBANK》, 13 May 2019 (2019-05-13) * |
| S E D\'SOUZA等: "Arginyl-glycyl-aspartic acid (RGD): a cell adhesion motif", 《TRENDS IN BIOCHEMICAL SCIENCES》, vol. 16, no. 07, 31 July 1991 (1991-07-31), pages 246 - 250, XP025864751, DOI: 10.1016/0968-0004(91)90096-E * |
| 孙云霞等: "不同链长RGD多肽对交联PEI体外细胞转染的影响", 《中国科技论文在线》, vol. 05, no. 06, 15 June 2010 (2010-06-15), pages 415 - 418 * |
| 杨大成、范莉、钟裕国: "全保护RGD三肽的合成方法研究", 《有机化学》, vol. 23, no. 05, 25 May 2003 (2003-05-25), pages 493 - 498 * |
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