Development of a Bestatin-SAHA Hybrid with Dual Inhibitory Activity against APN and HDAC
Abstract
:1. Introduction
2. Results and Discussion
2.1. Chemistry
2.2. In Vitro HDACs Inhibitory Assay
2.3. Western Blot Analysis
2.4. In Vitro APN Inhibitory Assay
2.5. Ex Vivo Rat Thoracic Aorta Rings (TARs) Assay
3. Materials and Methods
3.1. Chemistry
3.1.1. The Synthesis of Target Compound P1
((2S,3R)-3-((Tert-butoxycarbonyl)amino)-2-hydroxy-4-phenylbutanoyl)-l-leucine (14)
Tert-butyl (4-(6-(((benzyloxy)carbonyl)amino)hexanamido)phenyl)carbamate (16)
Benzyl (6-((4-aminophenyl)amino)-6-oxohexyl)carbamate (17)
Methyl 8-((4-(6-(((benzyloxy)carbonyl)amino)hexanamido)phenyl)amino)-8-oxooctanoate (18)
Methyl 8-((4-(6-aminohexanamido)phenyl)amino)-8-oxooctanoate (19)
Methyl 8-((4-((6R,7S,10S)-6-benzyl-7-hydroxy-10-isobutyl-2,2-dimethyl-4,8,11-trioxo-3-oxa-5,9,12-triazaoctadecan-18-amido)phenyl)amino)-8-oxooctanoate (20)
Tert-butyl ((2R,3S)-3-hydroxy-4-(((S)-1-((6-((4-(8-(hydroxyamino)-8-oxo-octanamido)phenyl) amino)-6-oxohexyl)amino)-4-methyl-1-oxopentan-2-yl)amino) -4-oxo-1-phenylbutan-2-yl)carbamate (21)
N1-(4-(6-((S)-2-((2S,3R)-3-Amino-2-hydroxy-4-phenylbutanamido)-4-methylpentanamido) hexanamido)phenyl)-N8-hydroxyoctanediamide trifluoroacetate (P1)
3.1.2. The Synthesis of Target Compound P2
Methyl 3-((2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)carbamoyl)-benzoate (23)
Methyl 3-((2-(2-(2-aminoethoxy)ethoxy)ethyl)carbamoyl)benzoate hydrochloride (24)
Methyl 3-(((6R,7S,10S)-6-benzyl-7-hydroxy-10-isobutyl-2,2-dimethyl-4,8,11-trioxo-3,15,18-trioxa-5,9,12-triazaicosan-20-yl)carbamoyl)benzoate (25)
Tert-butyl ((13S,16S,17R)-16-hydroxy-1-(3-(hydroxycarbamoyl)phenyl)-13-isobutyl-1,12,15-trioxo-18-phenyl-5,8-dioxa-2,11,14-triazaoctadecan-17-yl)carbamate (26)
N1-((11S,14S,15R)-15-Amino-14-hydroxy-11-isobutyl-10,13-dioxo-16-phenyl-3,6-dioxa-9,12-diazahexadecyl)-N3-hydroxyisophthalamide trifluoroacetate (P2)
3.1.3. The Synthesis of Target Compound P3
2-(2-(2-(2-Hydroxyethoxy)ethoxy)ethyl)isoindoline-1,3-dione (28)
2-(2-(2-(1,3-Dioxoisoindolin-2-yl)ethoxy)ethoxy)acetic acid (29)
2,2-Dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-oic acid (30)
3-Amino-N-(benzyloxy)benzamide (32)
3-(2-(2-(2-Aminoethoxy)ethoxy)acetamido)-N-(benzyloxy)benzamide hydrochloride (33)
Tert-butyl ((11S,14S,15R)-1-((3-((benzyloxy)carbamoyl)phenyl)amino)-14-hydroxy-11-isobutyl- 1,10,13-trioxo-16-phenyl-3,6-dioxa-9,12-diazahexadecan-15-yl)carbamate (34)
3-((11S,14S,15R)-15-Amino-14-hydroxy-11-isobutyl-10,13-dioxo-16-phenyl-3,6-dioxa-9,12- diazahexadecanamido)-N-hydroxybenzamide (P3)
3.2. In Vitro HDACs Inhibitory Assay
3.3. Western Blot Analysis
3.4. In Vitro APN Inhibitory Assay
3.5. Ex Vivo Rat Aortic Rings Assay
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
HDACs | histone deacetylases |
PROTACs | proteolysis-targeting chimeras |
VHL | von Hippel-Lindau |
cIAP1 | cellular inhibitor of apoptosis protein 1 |
SNIPERs | specific and nongenetic IAPs-dependent protein eraser |
CRABPs | cellular retinoic acid-binding proteins |
RARα | retinoic acid receptor α |
AR | androgen receptor |
ERα | estrogen receptor α |
APN | aminopeptidase N |
TARs | rat thoracic aorta rings |
ppm | parts per million |
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Compounds | Inhibitory Rates at 10 μM (IC50) a | ||
---|---|---|---|
HDAC1 | HDAC6 | HDAC8 | |
P1 | 98% (30.0 ± 2.8 nM) | 99% (11.5 ± 0.71 nM) | 90% (694.0 ± 110.3 nM) |
P2 | 66% | 76% | 64% |
P3 | 67% | 71% | 51% |
SAHA | 63.5 ± 3.5 nM | 21.0 ± 1.4 nM | >7000 nM |
| NT b | 2.5 ± 1.6 μM c | 14 ± 10 μM c |
| NT b | 0.65 ± 0.52 μM c | 3.3 ± 1.7 μM c |
Compounds | IC50 (μM) a |
---|---|
P1 | 3.43 ± 0.14 |
P2 | 1.67 ± 0.03 |
P3 | 2.65 ± 0.16 |
bestatin | 4.33 ± 0.50 |
Sample Availability: Samples of the compound P1 are available from the authors. |
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Share and Cite
Cao, J.; Zhao, W.; Zhao, C.; Liu, Q.; Li, S.; Zhang, G.; Chou, C.J.; Zhang, Y. Development of a Bestatin-SAHA Hybrid with Dual Inhibitory Activity against APN and HDAC. Molecules 2020, 25, 4991. https://doi.org/10.3390/molecules25214991
Cao J, Zhao W, Zhao C, Liu Q, Li S, Zhang G, Chou CJ, Zhang Y. Development of a Bestatin-SAHA Hybrid with Dual Inhibitory Activity against APN and HDAC. Molecules. 2020; 25(21):4991. https://doi.org/10.3390/molecules25214991
Chicago/Turabian StyleCao, Jiangying, Wei Zhao, Chunlong Zhao, Qian Liu, Shunda Li, Guozhen Zhang, C. James Chou, and Yingjie Zhang. 2020. "Development of a Bestatin-SAHA Hybrid with Dual Inhibitory Activity against APN and HDAC" Molecules 25, no. 21: 4991. https://doi.org/10.3390/molecules25214991