KR101482999B1 - Anti-inflammatory therapeutic Agents containg (E)-1-(2-(2-Nitrovinyl)Phenyl)Pyrrolidine as an Suppressor of TRIF-dependent signaling pathway of Toll-like receptors - Google Patents

Abstract

The present invention relates to the use of ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient as an inhibitor of the TRIF-dependent signaling pathway induced by toll- Lt; / RTI >
Toll-like receptors (TLRs) play an important role in recognizing many pathogenic molecular patterns and induce congenital immune responses. Recognition of bacterial factors by TLRs allows MyD88 and (TRIF) -dependent downstream signaling pathways to be initiated. As described above, the inventors synthesized ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) containing nitrovinyl-phenyl and pyrrolidine. To evaluate the potential of NVPP as a therapeutic agent, a signal through the TRIF-dependent pathway of TLRs induced by lipopolysaccharide (LPS), polyriboinosinic polyribosytidylic acid (Poly [I: C]) The effects of induction of delivery were investigated. NVPP not only inhibits the activation of interferon inducible genes such as interferon inducible protein-10 but also inhibits the activation of NF-kB, interferon regulatory factor and interferon regulatory factor 3 phosphorylation induced by LPS or Poly [I: C] do. These results suggest that NVPP may modulate the TRIF-dependent signaling pathways of TLRs and thus may be developed as an effective potential treatment for chronic inflammatory diseases.

Description

An anti-inflammatory agent comprising (E) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient as an inhibitor of TRIF-dependent signaling pathway induced by tol- {Anti-inflammatory therapeutic Agents Containing (E) -1- (2- (2-Nitrovinyl) Phenyl) Pyrrolidine as Suppressor of TRIF-dependent Signaling Pathway of Toll-

The present invention relates to the use of ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient as an inhibitor of the TRIF-dependent signaling pathway induced by toll- Lt; / RTI >

Congenital immunity is recognized as important for host defense against invading bacterial pathogens. Pattern recognition receptors (PRRs) are germ-line encoded receptors that recognize pathogen-associated molecular patterns (PAMPs), resulting in innate and acquired immune responses (Bjorkbacka et al., 2004; Medzhitov et al., 1997).

Well-studied PRRs are Toll-like receptors (TLRs). Activation of TLRs by PAMPs induces a variety of intercellular signaling cascades leading to the transcription of other genes important for the defense of cytokines, chemokines and hosts.

Generally, signaling through TLRs induces one or more toll / interleukin (IL) -1 receptor (TIR) domain-containing adapter molecules, namely myeloid differentiation factor 88 (MyD88) and inferfer- By adopting the Toll / IL-1R Domain-Incorporated Adapter (TRIF), it is possible to divide into two delivery paths (O'Neill and Bowie, 2007). These adapters facilitate activation of MyD88- and TRIF-dependent pathways.

Activation of TLR1, TLR2, TLR5, TLR6, TLR7, TLR8, TLR9 and TLR11 and their ligands adopt MyD88, whereas activation of TLR3 and its ligands adopts TRIF. TLR4 adopts both MyD88 and TRIF (Takeda and Akira, 2005).

The adoption of adapter molecules allows the initiation of cascade of signaling pathways and the activation of transcription factors such as nuclear factor-κB (NF-κB) and IFN regulatory factor 3 (IRF3), leading to inflammatory cytokines, chemokines ) And type I IFNs (Takeda and Akira, 2005).

 All TLR family members except TLR3 carry an inflammatory signal response through the conserved classical pathway (Takeda and Akira, 2005). This pathway is initiated by a conserved cytoplasmic protein sequence called the TIR domain, which binds to IL-1-associated kinase (IRAK) -1 and IRAK-4, TRAF-6, mitogen-activated protein kinases (MAPK) Activates the signaling mediator involved, activating the prototypic inflammatory transcription factor NF-kB.

These signaling pathways lead to the expression of inflammatory gene products, including cytokines (Akira et al., 2006). In addition to inflammatory signaling, some TLRs adopt Mal, TRAM and TRIF and confer specificity for signaling (Creagh and O'Neill, 2006).

TRIF has been shown to play an important role in signal transduction by TLR4 and TLR3, while TRAM has been shown to be essential for signaling only TLR4. TRIF induces the expression of IFN-inducible genes such as activation of IRF3 and regulated on activation normal T-cell expressed and secreted (RANTES) (Fitzgerald et al., 2003; Gao et al. 1998; Kawai et al., 2001).

( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) (Fig. 1 (A)) comprising nitrovinyl-phenyl and pyrrolidine is prepared by preparing a biologically useful molecule Was synthesized at our laboratory using building blocks.

As described above, the inventors have reported that NVPP inhibits the MyD88-dependent signaling pathway of TLRs (Eom et al., 2013). NVPP also inhibits NF-kB activation and iNOS expression induced by lipopolysaccharide (LPS; TLR4 agonist) and macrophage-activated lipopeptide 2-kDa (MALP-2; TLR2 and TLR6 agonists).

However, it is not known whether NVPP inhibits the TRIF-dependent signaling pathway of TLRs. Accordingly, the present invention seeks to identify an anti-inflammatory target of NVPP in the TRIF-dependent signaling pathway of TLRs.

The present invention relates to the use of ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine as an inhibitor of the TRIF-dependent signaling pathway induced by tol-like receptors (TLRs) NVPP) as an active ingredient.

In order to achieve the above object, the present invention provides an anti-inflammatory agent comprising ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient as an inhibitor of TRIF- Lt; / RTI >

In addition, the (E) -1- (2- (2- nitro-vinyl) phenyl) pyrrolidine in the present invention (NVPP) is (E) as an inhibitor of TRIF- dependent signaling having Formula 1 of the following - The present invention provides an anti-inflammatory agent comprising 1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient.

[Chemical Formula 1]

 Toll-like receptors (TLRs) play an important role in recognizing many pathogenic molecular patterns and induce congenital immune responses. Recognition of bacterial factors by TLRs allows MyD88 (myeloid differential factor 88) and toll-interleukin-1 receptor domain-containing adapter-initiated interferon-beta (TRIF) -dependent downstream signaling pathways to initiate.

As described above, the inventors synthesized ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) containing nitrovinyl-phenyl and pyrrolidine. To evaluate the potential of NVPP as a therapeutic agent, a signal through the TRIF-dependent pathway of TLRs induced by lipopolysaccharide (LPS), polyriboinosinic polyribosytidylic acid (Poly [I: C]) The effects of induction of delivery were investigated.

 NVPP not only inhibits the activation of interferon inducible genes such as interferon inducible protein-10 (IP-10) but also inhibits the activation of NF-κB, interferon regulatory factor and interferon regulatory factor 3 induced by LPS or Poly [I: Inhibits the activation of phosphorylation. These results suggest that NVPP may modulate the TRIF-dependent signaling pathways of TLRs and thus may be developed as an effective potential treatment for chronic inflammatory diseases.

Figure 1 shows the structure of ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP).
Figure 2 shows that NVPP inhibits the MyD88-dependent signaling pathway of TLR4. Figure 2 (A) shows that RAW264.7 cells were transformed with the NF- [kappa] B luciferase reporter plasmid, and NVPP (20 , 50, 100 [mu] M) and then treated with LPS (10 ng / ml) for an additional 8 hours.
Cell lysates were prepared and the activity of luciferase and β-gal enzyme was measured. Relative luciferase activity was normalized to? -Gal activity. Measurements were expressed as mean ± SEM (n = 3). (A) *, significantly different from LPS p < 0.05 (#). (BD) 293T cells were transformed with the NF-κB luciferase reporter plasmid and MyD88 (B), IKKβ (C) or p65 (D).
Cells were treated with NVPP (20, 50, 100 [mu] M) for an additional 18 hours. Relative luciferase activity was normalized to? -Gal activity. Measurements were expressed as mean ± SEM (n = 3). (B) + is significantly different from MyD88. p < 0.01 (++). (C) # is quite different from IKKβ. p < 0.01 (##). (D) ♣ indicates that the result is significantly different from p65. p < 0.01 (♣. Veh, vehicle; NVPP, ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine.
FIG. 3 shows that NVPP inhibits LPS-induced IRF3 activity. FIG. 3 (A) shows that RAW264.7 cells were transformed with IRF3 binding site (IFNβ PRDIII-I) (20, 50, 100 [mu] M) and then treated with LPS (10 ng / ml) for an additional 8 hours.
Cell lysates were prepared and the activity of luciferase and β-gal enzyme was measured. Relative luciferase activity was normalized to? -Gal activity. Measurements were expressed as mean ± SEM (n = 3). * Indicates significantly different from LPS. p < 0.01 (**).
FIG. 3 (B) shows RAW 264.7 cells treated with NVPP (20, 50, 100 μM) for 1 hour in advance and further treated with LPS (10 ng / ml) for 8 hours. Cell lysates were analyzed for pIRF3 and IRF3 proteins by Western blotting.
Figure 3 (C) shows that RAW264.7 cells are transformed with IP-10-luciferase reporter plasmids and treated with NVPP (20, 50, 100 [mu] M) for 1 hour in advance and LPS ). Cell lysates were prepared and the luciferase and beta-gal enzyme activities were measured. Relative luciferase activity was normalized to? -Gal activity. Measurements were expressed as mean ± SEM (n = 3). + Indicates significantly different from LPS. p < 0.05 (+), p < 0.01 (++).
Figure 3 (D) shows RAW 264.7 cells treated with NVPP (20, 50, 100 μM) for 1 hour in advance and stimulated with LPS (10 ng / ml) for an additional 8 hours. Cell lysates were analyzed for IP-10 and beta-actin proteins by Western blotting. Veh, vehicle; NVPP, ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine.
Figure 4 shows that NVPP inhibits the TRIF-dependent signaling pathway of TLR3. (AC) RAW264.7 cells were transformed with NF-κB (A), IRF3 binding site (IFNβ PRDIII-I) (B) and IP-10 (C) luciferase reporter plasmid, 20, 50, 100 μM) and treated with poly [I: C] (10 μg / ml) for an additional 8 hours.
Cell lysates were prepared and the luciferase and beta-gal enzyme activities were measured. Relative luciferase activity was normalized to? -Gal activity. Measurements were expressed as mean ± SEM (n = 3). (A) * indicates that the result is quite different from poly [I: C]. p < 0.05 (*), p < 0.01 (**). (B) + indicates that the result is significantly different from poly [I: C]. p < 0.01 (++). (C) # indicates that the result is significantly different from poly [I: C]. p < 0.05 (#), p < 0.01 (##). (D) RAW 264.7 cells were pretreated with NVPP (20, 50, 100 μM) for 1 hour and stimulated with poly [I: C] (10 μg / ml) for an additional 8 hours. Cell lysates were analyzed for IP-10 and beta-actin proteins by Western blotting. Veh, vehicle; NVPP, ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine.
Figure 5 shows inhibition of IRF3 activation by the NVPPrk downstream signaling component. (A) 293T cells express IRF3 binding site (IFN beta PRDIII-I) -luciferase plasmid and TRIF expression plasmid (A) TBK1 (B) or IRF3 5D (C). Cells were treated with NVPP (20, 50, 100 [mu] M) for 18 hours. Relative luciferase activity was normalized to? -Gal activity. Measurements were expressed as mean ± SEM (n = 3). Veh, vehicle; NVPP, ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine.

The present invention relates to the use of ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient as an inhibitor of the TRIF-dependent signaling pathway induced by toll- Lt; / RTI &gt;

Hereinafter, the present invention will be described in detail with reference to examples.

[ Example  1: chemical synthesis]

NVPP was synthesized by modification of established procedures (Rabong et al., 2008). An oven-dried 100 mL flask containing a stir bar was charged with 2- (pyrrolidin-1-yl) benzaldehyde (0.875 g, 5 mmol), KF (0.192 g, 0.327 mmol), Me ₂ NH ₂ Cl ), Nitromethane (11.2 mL, 203 mmol) and toluene (11 mL). The flask is mounted on a Dean-Stark apparatus and refluxed for 5 hours. The solution was removed under reduced pressure and the residue was purified by flash chromatography (ethyl acetate / hexane = 1/20) to give NVPP (0.9 g, 60%). Mp 175 ^° C; ¹ H NMR (200 MHz, CDCl ₃ ) d 8.33 (d, J = 13.4 Hz, 1H), 7.37 (d, J = 13.4 Hz, 1H), 7.30-7.17 (m, 2H), 6.83-6.66 2H), 3.30-3.23 (m, 4H), 1.94-1.85 (m, 4H). ¹³ C NMR (50 MHz) d 148.7, 137.1, 132.3, 130.1, 127.2, 116.7, 116.2, 113.5, 50.4, 23.2.

[ Example  2: Reagent ( reagents )]

LPS was purchased from List Biological Laboratories (San Jose, CA, USA). Poly [I: C] was purchased from Amersham Biosciences (Piscataway, NJ, USA). LPS and Poly [I: C] were dissolved in endotoxin-free water. All other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise noted.

[ Example  3: cell culture]

RAW264.7 cells (rat mononuclear leukocytes; ATTCC TIB71) and 293T human embryonic kidney cells were treated with 10% (v / v) heat-inactivated fetal bovine serum, 100 units / mL penicillin Invitrogen, Carlsbad, Calif., USA), and 100 μg / mL streptomycin (Invitrogen). Cells were maintained at 37 ° C in 5% CO ₂ atmosphere unless otherwise noted.

[ Example  4: plasmid]

The NF-κB (2x) -luciferase reporter construct was provided by Frank Mercurio (Signal Pharmaceuticals, CA). An IFN beta PRDIII-I-luciferase reporter plasmid was provided by Kate Fitzgerald (University of Massachusetts Medical School, MA). Heat shock protein 70 (HSP70) -β-galactosidase (β-gal) reporter plasmid was obtained from Robert Modlin (University of California, CA). The IP-10-Luciferase reporter construct was obtained from Daniel Hwang (University of California, CA). All DNA constructs were prepared on a large scale using the EndoFree Plasmid Maxi kit (Qiagen, Valencia, CA, USA) for transformation.

[ Example  5: Transformation and Luciferase  analysis]

Analysis was performed as described above (Ahn et al., 2009; Youn et al., 2009). Approximately, RAW264.7 cells were incubated with luciferase plasmid and heat shock protein (HSP) 70-β-galactosidase as internal controls using SuperFect transfection reagent (Qiagen, Valencia, CA, USA) Lt; RTI ID = 0.0 &gt; plasmid. &Lt; / RTI &gt; Luciferase and β-gal enzyme activity was measured using commercial luciferase and β-gal enzyme activity assay system (Promega) according to the manufacturer's instructions. The luciferase activity was normalized by? -Galactosidase activity.

[ Example  6: Western Blot  analysis( Western blot 분석 )]

Western blotting was performed as described above (Youn et al., 2006b; Yun et al., 2009). The same amount of cell extract was treated with 8% SDS gel electrophoresis (SDS-PAGE), and the separated proteins were electro-transferred to a polyvinylidene difluoride membrane. Membranes were blocked to remove non-specifically bound antibodies in a phosphate-buffered saline containing 0.1% Tween-20 and 3% skim milk powder. Immunoblotting was performed with the labeled antibody and secondary antibody conjugated with horseradish peroxidase (Amersham Biosciences, Arlington Heights, IL, USA). Reaction bands were visualized with an enhanced chemiluminescence Western blot detection reagent (Intron, Seongnam, Gyeonggi-do, South Korea). The membrane was resolved with 0.2 N NaOH for 10 min at room temperature to re-examine other antibodies.

[ Example  7: Statistical analysis]

Data were obtained from three experiments. Measurements were expressed as mean ± standard error (SEM). The error of the data was measured by the student's t test. P-values less than 0.05 were used to indicate statistically significant differences.

The result of the experiment through the above embodiment is analyzed as follows.

NVPP inhibits the MyD88-dependent signaling pathway of TLRs. In general, the TLRs signaling pathways consist of MyD88- and TRIF-dependent pathways. Both MyD88- and TRIF-dependent pathways can lead to NF-κB activation, and NF-κB is a common downstream signaling component of TLRs. TLR4 initiates activation of both MyD88- and TRIF-dependent pathways. Thus, NF-kB activation induced by LPS (TLR4 agonist) is used to determine whether TLRs are activated. NVPP inhibits LPS induced NF- [kappa] B activation in a dose-dependent manner as confirmed in the luciferase reporter gene assay of Fig. 2 (A).

In order to investigate the regulation of the MyD88-dependent signaling pathway by NVPP, NF-κB activation was induced by overexpression of MyD88, IKKb, or p65 in 293T cells. NVPP inhibits agonist-independent activation induced by MyD88 (Figure 2 (B)), IKKb (Figure 2 (C)) or p65 (Figure 2 (D)), indicating that NVPP inhibits MyD88- It is to prove that the path is suppressed.

In addition, NVPP inhibits the TRIF-dependent signaling pathway of TLR4. The inventors investigated whether NVPP inhibits the TRIF-dependent signaling pathway of TLR4. Since the TRIF-dependent signaling pathway leads to the activation of the transcription factor IRF3 (Fitzgerald et al., 2003), IRF3 activation is used to read the TRIF-dependent signaling pathway. As shown by the reporter gene analysis using the IFN beta promoter domain containing the IRF3 binding site (IFN beta PRDIII-I) (Fig. 3 (B)), NVPP inhibits LPS-induced IRF3 activation. In addition, NVPP inhibits the phosphorylation of IRF3 as confirmed in Western blotting (Fig. 3 (B)).

To determine whether NVPP can regulate TRIF-dependent pathways, the expression of genes associated with TRIF-dependent pathways such as IP-10 was determined by luciferase reporter gene analysis and Western blotting. NVPP inhibits LPS-induced IP-10 expression, as confirmed in the luciferase reporter gene assay (Figure 3 (C)) and Western blotting (Figure 3 (D)). These results suggest that NVPP inhibits the TRIF-dependent signaling pathway derived from TLR4 activation.

In addition, NVPP inhibits the TRIF-dependent signaling pathway of TLR3. Although the TLR4 signaling pathway may be initiated by activation of transcription factors mediated by both the MyD88- and TRIF-dependent pathways, the TLR3 signaling pathway is only required to initiate transcription factor activation by the TRIF-dependent signaling pathway do. Thus, induction of NF-κB and IRF3 activation by poly [I: C] can be used as a reading of the TRIF-dependent signaling pathway. NVPP inhibits poly [I: C] -induced NF-kB and IRF3 activation, as confirmed in the luciferase reporter gene assay (Fig. 4 (A) and 4 (B)). NVPP also inhibits poly [I: C] -induced IP-10 expression, as confirmed in luciferase reporter gene analysis (Fig. 4C) and Western blotting (Fig. 4D). These results indicate that NVPP inhibits the TRIF-dependent signaling pathway derived from TLR3 activation.

In addition, NVPP does not inhibit the activation of IRF3 induced by the downstream signaling component of the TRIF-dependent signaling pathway of TLRs. Presently, to identify the molecular target of NVPP for inhibition of the TRIF-dependent signaling pathway, the downstream component of the pathway (TRIF, TBKl or IRF3) was transformed into 293T cells. The TRIF-dependent signaling pathway of TLRs causes IRF3 activation mediated through TRIF and TBK1. NVPP is an IRF3 activated by TRIF, TBK1 or IRF3 5D (a generic activated form of IRF3) as shown in the IRF3 binding site (IFNb PRDIII-I) reporter gene analysis (Fig. 5 .

These results suggest that the target of NVPP is not a TRIF-dependent downstream signaling component including the adapter molecule itself. The target may be an upstream component of the adapter molecule, including that causing TLRs activation by TLRs or agonists.

The MyD88- and TRIF-dependent signaling pathways are the two major TLRs downstream signaling pathways that activate the transcription factors NF-KB and IRFs. Both NF-κB and IRFs are simultaneously activated for bacterial infection, but the target genes induced by activation by NF-κB and IRFs are different.

NF-κB activation induces proinflammatory cytokines, whereas activation of IRFs induces type I IFN genes (Honda and Taniguchi, 2006; Stetson and Medzhitov, 2006; Wietek and O'Neill, 2007).

The Type I IFN gene is a key gene in establishing innate antiviral responses, and is activated by the cooperative action of NF-κB and IRF3. TLR3 and TLR4 activation induces the expression of genes encoding IFN and chemokines (Servant et al., 2002). NF-κB and IRF3-induced signaling pathways are initiated by the combination of pathogen-specific products and TLRs in macrophages and dendritic cells (Uematsu and Akira, 2007). TLRs are an important component of the host's immune system, sensing key microorganisms including bacteria, viruses and fungi.

Abnormalities in the regulation of TLRs-mediated cell responses can lead to chronic inflammation, which in turn affects the progression and development of many inflammatory diseases. Much evidence suggests that the TRIF-dependent signaling pathways of TLRs are important in inflammatory responses and certain chronic diseases. Expression of the majority of LPS-induced genes (> 70%) is mediated through TRIF-dependent signaling pathways (Schafer et al., 1998).

Thus, regulation of the TRIF-dependent signaling pathways of TLRs is important as an anti-inflammatory strategy. This demonstrates that inhibition of the TRIF-dependent signaling pathway by NVPP and inhibition of subsequent IRF3 activation can significantly inhibit target gene expression in the TLRs signaling pathway.

TRIF is an important adapter molecule that facilitates the activation of the TLR3 and TLR4 signaling pathways (O'Neill, 2008). These report results indicate that the TRIF signaling pathway is negatively regulated by the majority of molecules; On the other hand, in most cases they do not specifically target TRIF but affect downstream components of the TRIF pathway, such as TBK1 (Park and Youn, 2010; Youn et al., 2005; Youn et al., 2006a ).

We have demonstrated that many phytochemicals with α, β-unsaturated carbonyl group structure motifs that have undergone Michael addition have been shown to inhibit TLR4 dimer formation (Ahn et al., 2009 ; Youn et al., 2008; Youn et al., 2006b). It has been reported that molecules with such structural motifs exhibit very high reactivity with the sulfhydryl group of cysteine by the Michael addition reaction.

TLRs have several cysteine residues in the extracellular and intracellular domains that may be involved in the formation of disulfide bonds for dimerization of the receptor. NVPP also has an alpha, beta -unsaturated nitro group structure motif with the potential for Michael addition reactions. This indicates that NVPP can react with the cysteine residues in TLR4 causing inhibition of TLR4 dimer formation. Future studies will confirm the exact molecular target of NVPP in the TLRs signaling pathway.

In the present invention, the inventors have demonstrated that NVPP inhibits the TRIF-dependent pathway of TLR3 and TLR4. Inhibition of the TRIF pathway of TLR3 and TLR4 by NVPP is accompanied by inhibition of activation of NF-kB and IRF3, including IFN beta and IP-10, and their target genes.

All these results indicate a significant possibility that the risk for TLR4-mediated inflammatory response and thus many chronic inflammatory responses can be controlled by the nitrovinyl derivative NVPP.

The above-described embodiments are illustrative and those skilled in the art will be able to make various modifications and alterations to the disclosed embodiments without departing from the spirit of the present invention. The scope of the present invention is not limited by these variations and modifications.

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Claims (2)

An anti-inflammatory agent comprising ( E ) -1- (2- (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient as an inhibitor of the TRIF-dependent signaling pathway. The method according to claim 1,
The (E) -1- (2- (2- nitro-vinyl) phenyl) pyrrolidine (NVPP) is (E) -1- (2- TRIF- as inhibitors of dependent signaling with Formula 1 below (2-nitrovinyl) phenyl) pyrrolidine (NVPP) as an active ingredient.
[Chemical Formula 1]

Images