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In Silico Identification and In Vitro and In Vivo Validation of Anti-Psychotic Drug Fluspirilene as a Potential CDK2 Inhibitor and a Candidate Anti-Cancer Drug.

Shi XN, Li H, Yao H, Liu X, Li L, Leung KS, Kung HF, Lu D, Wong MH, Lin MC - PLoS ONE (2015)

Bottom Line: Our results showed that oral fluspirilene treatment significantly inhibited tumor growth.Moreover, the cocktail treatment with fluspirilene and 5-fluorouracil exhibited the highest therapeutic effect.In view of the fact that fluspirilene has a long history of safe human use, our discovery of fluspirilene as a potential anti-HCC drug may present an immediately applicable clinical therapy.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Center, Kunming Medical University, Kunming, Yunnan, China; Department of Medicine, Southwest Guizhou Vocational and Technical College for Nationalities, Guizhou, China.

ABSTRACT
Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. Surgical resection and conventional chemotherapy and radiotherapy ultimately fail due to tumor recurrence and HCC's resistance. The development of novel therapies against HCC is thus urgently required. The cyclin-dependent kinase (CDK) pathways are important and well-established targets for cancer treatment. In particular, CDK2 is a key factor regulating the cell cycle G1 to S transition and a hallmark for cancers. In this study, we utilized our free and open-source protein-ligand docking software, idock, prospectively to identify potential CDK2 inhibitors from 4,311 FDA-approved small molecule drugs using a repurposing strategy and an ensemble docking methodology. Sorted by average idock score, nine compounds were purchased and tested in vitro. Among them, the anti-psychotic drug fluspirilene exhibited the highest anti-proliferative effect in human hepatocellular carcinoma HepG2 and Huh7 cells. We demonstrated for the first time that fluspirilene treatment significantly increased the percentage of cells in G1 phase, and decreased the expressions of CDK2, cyclin E and Rb, as well as the phosphorylations of CDK2 on Thr160 and Rb on Ser795. We also examined the anti-cancer effect of fluspirilene in vivo in BALB/C nude mice subcutaneously xenografted with human hepatocellular carcinoma Huh7 cells. Our results showed that oral fluspirilene treatment significantly inhibited tumor growth. Fluspirilene (15 mg/kg) exhibited strong anti-tumor activity, comparable to that of the leading cancer drug 5-fluorouracil (10 mg/kg). Moreover, the cocktail treatment with fluspirilene and 5-fluorouracil exhibited the highest therapeutic effect. These results suggested for the first time that fluspirilene is a potential CDK2 inhibitor and a candidate anti-cancer drug for the treatment of human hepatocellular carcinoma. In view of the fact that fluspirilene has a long history of safe human use, our discovery of fluspirilene as a potential anti-HCC drug may present an immediately applicable clinical therapy.

No MeSH data available.


Related in: MedlinePlus

The predicted conformation of fluspirilene in complex with CDK2.CDK2 residues are rendered as lines colored by atom type. Fluspirilene is rendered as sticks colored by atom type. The interacting atoms and residues are labeled. The cyan, green and pink dashed lines represent hydrogen bonds, hydrophobic contacts and π interactions, respectively. This figure was created by iview [16].
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pone.0132072.g009: The predicted conformation of fluspirilene in complex with CDK2.CDK2 residues are rendered as lines colored by atom type. Fluspirilene is rendered as sticks colored by atom type. The interacting atoms and residues are labeled. The cyan, green and pink dashed lines represent hydrogen bonds, hydrophobic contacts and π interactions, respectively. This figure was created by iview [16].

Mentions: Fig 9 plots the predicted conformation of fluspirilene in complex with CDK2 (PDB ID: 1GZ8) using iview [16]. Fluspirilene was predicted to bind inside the ATP-binding pocket of CDK2 and interact with CDK2 mainly through hydrogen bonds, hydrophobic contacts and cation-π interactions. Putatively, the H8 atom forms a hydrogen bond with the backbone oxygen of Ile10, and the H32 atom forms another two hydrogen bonds with the backbone oxygen of Leu83 and His84. The side chains of Ile10, Ala31 and Leu134 establish a hydrophobic tunnel for the hydrophobic tail of fluspirilene to bury inside. One of the two aromatic rings located at the tail of fluspirilene forms a cation-π interaction with the positively charged side chain of Lys33. All these putative hydrogen bonds, hydrophobic contacts and cation-π interactions are spread over the head, middle and tail fragments of fluspirilene, thereby firmly holding fluspirilene at its predicted position and orientation. In comparison, the known CDK2 inhibitor 2-Amino-6-(3’-methyl-2’-oxo)butoxypurine (codenamed MBP for short), which is an O(6)-substituted guanine derivative, is known to establish four hydrogen bonds with Lys33, Glu81 and Leu83 (S2 Fig).


In Silico Identification and In Vitro and In Vivo Validation of Anti-Psychotic Drug Fluspirilene as a Potential CDK2 Inhibitor and a Candidate Anti-Cancer Drug.

Shi XN, Li H, Yao H, Liu X, Li L, Leung KS, Kung HF, Lu D, Wong MH, Lin MC - PLoS ONE (2015)

The predicted conformation of fluspirilene in complex with CDK2.CDK2 residues are rendered as lines colored by atom type. Fluspirilene is rendered as sticks colored by atom type. The interacting atoms and residues are labeled. The cyan, green and pink dashed lines represent hydrogen bonds, hydrophobic contacts and π interactions, respectively. This figure was created by iview [16].
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493148&req=5

pone.0132072.g009: The predicted conformation of fluspirilene in complex with CDK2.CDK2 residues are rendered as lines colored by atom type. Fluspirilene is rendered as sticks colored by atom type. The interacting atoms and residues are labeled. The cyan, green and pink dashed lines represent hydrogen bonds, hydrophobic contacts and π interactions, respectively. This figure was created by iview [16].
Mentions: Fig 9 plots the predicted conformation of fluspirilene in complex with CDK2 (PDB ID: 1GZ8) using iview [16]. Fluspirilene was predicted to bind inside the ATP-binding pocket of CDK2 and interact with CDK2 mainly through hydrogen bonds, hydrophobic contacts and cation-π interactions. Putatively, the H8 atom forms a hydrogen bond with the backbone oxygen of Ile10, and the H32 atom forms another two hydrogen bonds with the backbone oxygen of Leu83 and His84. The side chains of Ile10, Ala31 and Leu134 establish a hydrophobic tunnel for the hydrophobic tail of fluspirilene to bury inside. One of the two aromatic rings located at the tail of fluspirilene forms a cation-π interaction with the positively charged side chain of Lys33. All these putative hydrogen bonds, hydrophobic contacts and cation-π interactions are spread over the head, middle and tail fragments of fluspirilene, thereby firmly holding fluspirilene at its predicted position and orientation. In comparison, the known CDK2 inhibitor 2-Amino-6-(3’-methyl-2’-oxo)butoxypurine (codenamed MBP for short), which is an O(6)-substituted guanine derivative, is known to establish four hydrogen bonds with Lys33, Glu81 and Leu83 (S2 Fig).

Bottom Line: Our results showed that oral fluspirilene treatment significantly inhibited tumor growth.Moreover, the cocktail treatment with fluspirilene and 5-fluorouracil exhibited the highest therapeutic effect.In view of the fact that fluspirilene has a long history of safe human use, our discovery of fluspirilene as a potential anti-HCC drug may present an immediately applicable clinical therapy.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Center, Kunming Medical University, Kunming, Yunnan, China; Department of Medicine, Southwest Guizhou Vocational and Technical College for Nationalities, Guizhou, China.

ABSTRACT
Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. Surgical resection and conventional chemotherapy and radiotherapy ultimately fail due to tumor recurrence and HCC's resistance. The development of novel therapies against HCC is thus urgently required. The cyclin-dependent kinase (CDK) pathways are important and well-established targets for cancer treatment. In particular, CDK2 is a key factor regulating the cell cycle G1 to S transition and a hallmark for cancers. In this study, we utilized our free and open-source protein-ligand docking software, idock, prospectively to identify potential CDK2 inhibitors from 4,311 FDA-approved small molecule drugs using a repurposing strategy and an ensemble docking methodology. Sorted by average idock score, nine compounds were purchased and tested in vitro. Among them, the anti-psychotic drug fluspirilene exhibited the highest anti-proliferative effect in human hepatocellular carcinoma HepG2 and Huh7 cells. We demonstrated for the first time that fluspirilene treatment significantly increased the percentage of cells in G1 phase, and decreased the expressions of CDK2, cyclin E and Rb, as well as the phosphorylations of CDK2 on Thr160 and Rb on Ser795. We also examined the anti-cancer effect of fluspirilene in vivo in BALB/C nude mice subcutaneously xenografted with human hepatocellular carcinoma Huh7 cells. Our results showed that oral fluspirilene treatment significantly inhibited tumor growth. Fluspirilene (15 mg/kg) exhibited strong anti-tumor activity, comparable to that of the leading cancer drug 5-fluorouracil (10 mg/kg). Moreover, the cocktail treatment with fluspirilene and 5-fluorouracil exhibited the highest therapeutic effect. These results suggested for the first time that fluspirilene is a potential CDK2 inhibitor and a candidate anti-cancer drug for the treatment of human hepatocellular carcinoma. In view of the fact that fluspirilene has a long history of safe human use, our discovery of fluspirilene as a potential anti-HCC drug may present an immediately applicable clinical therapy.

No MeSH data available.


Related in: MedlinePlus