The effect of epidermal growth factor (EGF) conjugated with low-molecular-weight protamine (LMWP) on wound healing of the skin.
This study was designed to develop a skin permeable recombinant low-molecular-weight protamine (LMWP) conjugated epidermal growth factor (EGF) (rLMWP-EGF) by linking a highly positive charged LMWP to the N-terminal of EGF through genetic recombination. We evaluated its biological activity, skin permeability, and wound healing efficacy in vivo. The cDNA for rLMWP-EGF was prepared by serial polymerase chain reaction for encoding amino acids of LMWP to the vector for EGF. After expression and purification, recombinant EGF site-specifically conjugated with LMWP was obtained. The in vitro cell proliferation activity was well preserved after LMWP conjugation and was comparable to that of rEGF. rLMWP-EGF showed markedly improved permeability through the three-dimensional artificial human skin constructs, and the cumulative permeation of rLMWP-EGF across the excised mouse skin was about 11 times higher than that of rEGF. Topically applied rLMWP-EGF significantly accelerated the wound closure rate in full thickness as well as a diabetic wound model most probably due to its enhanced skin permeation. These findings demonstrate the therapeutic potential of rLMWP-EGF as a new topical wound healing drug and the site-specific conjugation of LMWP to peptides or proteins by genetic recombination as a useful method for preparing highly effective biomedicines.
Amorepacific Corporation R&D Center, 314-1, Bora-dong, Giheung-gu, Yongin-si, Gyeonggi-do 446-729, South Korea.
Jin Kyu
JK
Choi
Ji-Hye
JH
Jang
Won-Hee
WH
Jang
Jaekwan
J
Kim
Il-Hong
IH
Bae
Joonho
J
Bae
Young-Ho
YH
Park
Beum Joon
BJ
Kim
Kyung-Min
KM
Lim
Jin Woo
JW
Park
D000818:Animals; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000627:therapeutic use*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D011479:Protamines / Q000737:chemistry*; D012867:Skin / Q000187:drug effects* / Q000473:pathology*; D014945:Wound Healing / Q000187:drug effects*
D011479:Protamines; D004815:Epidermal Growth Factor