Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative
Autoři:
Fuding Chen aff001; Gang Du aff001; Meishu Shih aff002; Hongjiang Yuan aff002; Peng Bao aff001; Sheng Shi aff001; Yong Cang aff001; Zhen Zhang aff001
Působiště autorů:
Research and Development Division, Rejuven Dermaceutical Co., Ltd., Hangzhou, China
aff001; PharmaLegacy Laboratories (Shanghai) Co., Ltd., Shanghai, China
aff002
Vyšlo v časopise:
PLoS ONE 14(12)
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pone.0227202
Souhrn
Adipocytes attached to the extracellular matrix (ECM) mainly consist of collagen in adipose tissues, while the degradation of ECM by collagenase induces the apoptosis of adipocytes, leading to a decrease in local subcutaneous adipose. To achieve this goal, we are developing a mutant collagenase H (ColH) to remove local subcutaneous fat such as submental fat (SMF). Three vectors were constructed for expressing rColH(FM, mutant for fat melting, with 6xHis tag), rColH(WT, wild-type, with 6xHis tag), and rColH(E451D, E451D mutant, without 6xHis tag) in Escherichia coli. rColH(FM) & rColH(WT) were purified by Ni Sepharose on a laboratory scale, while rColH(E451D) was purified by five chromatography purification steps on a large scale. Then, the stability of rColH(FM) and rColH(WT) was tested by SDS-PAGE to investigate the influence of the E451D mutation on stability. Afterwards, the enzyme kinetics of ColH (mutant or wild-type, with or without His tag) were investigated and compared. Finally, the adipolysis of rColH(E451D) at various doses was tested in vitro and in vivo. The ultrasound results in minipigs suggested that effective adipolysis was induced by rColH(E451D) compared with the negative control, and the histological results suggest dose-dependent fibrosis, necrosis, inflammation and cholesterol cleft formation. These findings indicate the possibility of rColH(E451D) becoming a new injectable drug to safely remove subcutaneous adipose.
Klíčová slova:
Collagens – Fibrosis – Ultrasound imaging – Adipocytes – Enzyme kinetics – Collagenases – Lipolysis – Pharmacodynamics
Zdroje
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