Development and Structural Evaluation of N-alkylated phenylcyclopropylamine-based LSD1 Inhibitors.
Lysine-specific demethylase 1 (LSD1) is a flavin adenine dinucleotide (FAD)-dependent enzyme that catalyzes the demethylation of histone H3 and regulates gene expression. Because it is implicated in the regulation of diseases such as acute myeloid leukemia, potent LSD1-specific inhibitors have been pursued. Trans -2-phenylcyclopropylamine (2-PCPA)-based inhibitors featuring substitutions on the amino group have emerged, with submicromolar affinities toward LSD1 and high selectivities over monoamine oxidases (MAOs).
We synthesized two N -alkylated 2-PCPA-based LSD1 inhibitors, S2116 and S2157, based on the previously developed S2101. S2116 and S2157 exhibited enhanced potency for LSD1 by 2.0- to 2.6-fold, as compared with S2101. In addition, they exhibited improved selectivity over MAOs.
Structural analyses of LSD1 co-crystallized with S2101, S2116, S2157, or another N -alkylated inhibitor (FCPA-MPE) confirmed that the N -substituents enhance the potency of a 2-PCPA-based inhibitor of LSD1, without constituting the adduct formed with FAD.
Exogenous application of histone demethylase inhibitor mimics FLD loss-of-function phenotype in terms of systemic acquired resistance in Arabidopsis thaliana.
Plants often learn from previous infections to mount higher level of resistance during subsequent infections, a phenomenon referred to as systemic acquired resistance (SAR). During primary infection, mobile signals generated at the infection site subsequently move to the rest of plant to activate SAR. SAR activation is associated with alteration in the nucleosomal composition at the promoters of several defense-related genes.
However, genetic regulations of such epigenetic modifications are largely obscure. Recently, we have demonstrated that Reduced Systemic immunity1/FLOWERING LOCUS D (RSI1; alias FLD) a homolog of human histone demethylase, is required for SAR development in Arabidopsis.
Here, we report that exogenous application of a histone demethylase inhibitor trans-2-phenylcyclopropylamine (2-PCPA) mimics rsi1/fld loss-of-function phenotypes in terms of SAR and associated histone demethylation at the promoters of PR1, WRKY 29, and WRKY6 genes, and as well as flowering phenotypes. Our results suggest histone demethylase activity of FLD is important for controlling SAR activation.
The antidepressant phenylcyclopropylamine protects mice from high-fat-diet-induced obesity
Mice treated with the antidepressant trans-2-phenylcyclopropylamine (2-PCPA) were protected against diet-induced-obesity, and adiposity was reversed in pre-established diet-induced obese mice.
Contrary to a recent report that inhibition of lysine-specific demethylase-1 by 2-PCPA results in increased energy expenditure, long-term 2-PCPA treatment had no such effect but its protection against obesity was associated with increased spontaneous locomotor activity, Moreover, pair feeding to assure equal caloric intake in wild type mice as well as in genetic hyperphagic mice (ob/ob) also resulted in weight reduction in 2-PCPA treated mice that correlated with increased activity but no change in energy expenditure.
Similarly, short-term intraperitoneal injections of 2-PCPA did not affect food intake but caused a substantial increase in locomotor activity in the light cycle that correlated with increased energy expenditure, whereas activity and energy expenditure were unchanged in the dark cycle. Lastly, 2-PCPA was also effective in reducing obesity in genetic UCP1 null mice.
These data suggest that 2-PCPA can reduce obesity by decreasing food intake in the long term while increasing activity in the short-term. However, the protective and weight loss effects of 2-PCPA are independent of UCP1-regulated thermogenesis or basal energy expenditure.
phenylcyclopropylamine regulates zebrafish lateral line neuromast development mediated by depression of LSD1 activity.
The zebrafish mechanosensory lateral line (LL) is a model system for the study of hair cell development, survival and regeneration. Recently, histone modifications have attracted a considerable amount of interest because of their indispensable roles in various kinds of cellular processes including differentiation, proliferation, apoptosis and function.
Lysine specific demethylase 1 (LSD1) is an important enzyme that regulates histone methylation.
As a transcriptional regulator, this enzyme has broad functional activities and is involved in many biological processes. However, the effects of LSD1 on the early development of zebrafish sensory system have not been fully elucidated.
Here, we have found that pharmacological inhibition of LSD1 with the monoamine oxidase (MAO) inhibitor trans-2-phenylcyclopropylamine (referred to as 2-PCPA) reduced the numbers of both sensory hair cells and supporting cells of neuromasts during zebrafish development. Our results showed that the treatment of zebrafish larvae with 2-PCPA caused accumulation of histone methylation and suppressed proliferation of neuromast cells.
Finally, acridine orange staining assay demonstrated that 2-PCPA treatment at high concentrations induced an enhancement of cellular apoptosis within neuromasts. Taken together, these results indicate that LSD1 demethylase activity is required for neuromast development in zebrafish larvae.
Structurally designed phenylcyclopropylamine derivatives potently inhibit histone demethylase LSD1/KDM1 .
Lysine-specific demethylase 1 (LSD1/KDM1) demethylates histone H3, in addition to tumor suppressor p53 and DNA methyltransferase 1 (Dnmt1), thus regulating eukaryotic gene expression by altering chromatin structure. Specific inhibitors of LSD1 are desired as anticancer agents because LSD1 aberrations are associated with several cancers, and LSD1 inhibition restores the expression of abnormally silenced genes in cancerous cells.
In this study, we designed and synthesized several candidate compounds to inhibit LSD1, based on the structures of LSD1 and monoamine oxidase B (MAO-B), in complex with an antidepressant tranylcypromine (2-PCPA) derivative.
Compound S2101 exhibited stronger LSD1 inhibition than tranylcypromine and the known small LSD1 inhibitors in LSD1 demethylation assays, with a k(inact)/K(I) value of 4560 M(-1) s(-1). In comparison with tranylcypromine, the compound displayed weaker inhibition to the monoamine oxidases.
The inhibition modes of the two 2-PCPA derivatives, 2-PFPA and S1201, were identified by determination of the inhibitor-bound LSD1 structures, which revealed the enhanced stability of the inhibitor-FAD adducts by their interactions with the surrounding LSD1 residues. These molecules are potential pharmaceutical candidates for cancer or latent virus infection, as well as research tools for LSD1-related biological investigations.
phenylcyclopropylamine induces nerve cells apoptosis in zebrafish mediated by depression of LSD1 activity.
Trans-2-phenylcyclopropylamine (referred to as PCPA hereafter, also known as tranylcypromine and Parnate) is used clinically as an antidepressant. Here, we use a new model-zebrafish (Danio rerio) to study the molecular mechanisms of its adverse reactions in vivo.
Following a PCPA exposure (75 microM), embryos showed “sluggish” action (slow swim and slow escape action). Whole mount in situ hybridization showed that sox1a and huc expressions were downregulated in PCPA-treated embryos, which indicated a decrease in the number of nerve cells. TUNEL assay diplayed that the drop of nerve cells number due to excessive apoptosis.
trans 2-Phenylcyclopropylamine Hydrochloride (Tranylcypromine, trans 2-Amino-1-phenyl-cyclopropane Hydrochloride) |
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| MBS6038502-1g | MyBiosource | 1(g | 525 EUR |
trans 2-Phenylcyclopropylamine Hydrochloride (Tranylcypromine, trans 2-Amino-1-phenyl-cyclopropane Hydrochloride) |
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| MBS6038502-5x1g | MyBiosource | 5x1g | 2210 EUR |
(2-Phenylcyclopropyl)boronic Acid |
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| R706255 | Toronto Research Chemicals | 250mg | 1800 EUR |
N-[(1S,2R)-2-Phenylcyclopropyl]-acetamide |
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| P116400 | Toronto Research Chemicals | 500mg | 800 EUR |
1-(1-Phenylcyclopropyl)Piperazine |
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| abx181474-1g | Abbexa | 1 g | 777.6 EUR |
2-(1-Phenylcyclopropyl)-4-oxazolecarboxylic Acid |
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| P319785 | Toronto Research Chemicals | 250mg | 4500 EUR |
Potassium (1-Phenylcyclopropyl)trifluoroborate |
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| P698400 | Toronto Research Chemicals | 100mg | 1800 EUR |
Tranylcypromine Hemisulfate (Trans (+/-)-2-Phenylcyclopropanamine, Hemisulfate; SKF trans-355; 2-PCPA) |
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| MBS6088154-25mg | MyBiosource | 25(mg | 345 EUR |
Tranylcypromine Hemisulfate (Trans (+/-)-2-Phenylcyclopropanamine, Hemisulfate; SKF trans-355; 2-PCPA) |
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| MBS6088154-5x25mg | MyBiosource | 5x25mg | 1410 EUR |
Ethyl 2-Phenylcyclopropanecarboxylate |
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| 20-abx185416 | Abbexa |
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(1S,2S)-2-Phenylcyclopropane-1-carboxylic acid |
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| abx184838-1g | Abbexa | 1 g | 1762.8 EUR |
2-(Aminomethyl)-1-phenylcyclopropanecarboxamide |
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| A633435 | Toronto Research Chemicals | 100mg | 1800 EUR |
2-(Hydroxymethyl)-1-phenylcyclopropanecarbonitrile |
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| H972770 | Toronto Research Chemicals | 250mg | 3000 EUR |
(1S,2S)-2-(Hydroxymethyl)-1-phenylcyclopropanecarboxylic Acid |
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| H949640 | Toronto Research Chemicals | 25mg | 1800 EUR |
(1R,2R)-2-(Hydroxymethyl)-1-phenylcyclopropanecarboxylic Acid |
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| H949645 | Toronto Research Chemicals | 25mg | 1800 EUR |
1-Phenylcyclopropanesulfonyl Chloride |
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| P319965 | Toronto Research Chemicals | 1g | 3000 EUR |
Moreover, lysine-specific demethylase 1 morpholino injection (LSD1 MO) also induced increased cellular apoptosis in embryos just as PCPA. RT-PCR at 24hpf evaluated that the absence of LSD1 resulted in increased expression of two p53 target genes (p21 and bax2).
These findings demonstrate for the first time that PCPA-induced apoptosis through inhibition of LSD1 demethylase activity and p53-dependent signaling pathway might be required for the maintenance of nerve cell apoptosis.