The invention discloses a biscyclopeptide E(c(RGDfK))2 and a preparation method thereof. The method comprises the following steps: 1) immersing resin and Fmoc-Asp-oall (Fmoc-L-
aspartic acid alpha-allyl ester )in DCM (
dichloromethane), adding DIEA (N,N-diisopropylethylamine), carrying out a
contact reaction, and washing the obtained
reaction product; 2, adding DCM,
methanol and DIEA to the above obtained
system, carrying out end capping treatment, and adding
piperidine to carry out deprotection; 3) adding G (
glycine), HoBt (1-
hydroxybenzotriazole) and TBTU (2-(1H-
benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate) to the
reaction system, and carrying out a
contact reaction; 4) sequentially adding R (
arginine), K (
lysine) and f (
phenylalanine) into the
reaction system, carrying out a
contact reaction, adding Pd(PPh3)4, carrying out a
propenyl group removal reaction, and adding
piperidine to carry out deprotection; 5) adding DIEA, HoBt and HBTU into the
system, carrying out a contact reaction, adding a hydrazine
hydrate and DMF
mixed solution, carrying out a secondary reaction, and washing the obtained
reaction product to remove Dde; 6) dropwise adding Fmoc-Glu-OH, HoBt, DMF and TBTU to the
reaction system, carrying out a contact reaction, and washing the obtained
reaction product; and 7) adding
piperidine to the reaction
system to carry out deprotection, washing the obtained product with DMF, DCM and
methanol, and carrying out post-treatment to obtain the biscyclopeptide E(c(RGDfK))2. The biscyclopeptide E(c(RGDfK))2 has excellent stability.