Novel furfurylidene N-acylhydrazones derived from natural safrole: discovery of LASSBio-1215, a new potent antiplatelet prototype

Figures & data

Figure 1.  Currently available platelet antiaggregating drugs.

Figure 2.  Design concept of new functionalized furylydene 1,3-benzodioxolyl-N-acylhydrazine derivatives 9a–f and the vinylogous analogues 9g–j.

Scheme 1.  Reaction conditions: a) I₂, KOH, MeOH, rt, 6 h, 90%; b) NH₂NH₂.H₂O 80%, EtOH, reflux, 3.5 h, 84%; c) Functionalized 2-furyl-(CH=CH)_n-CHO (14a–e), EtOH, rt, 30 min, see Table 1; d) K₂CO₃, acetone, MeI, 40°C, 24 h, see supplementary Table S1.

Table 1.  Effect of functionalized 2-furyl N-acylhydrazone derivatives 9a–j on platelet aggregation induced by arachidonic acid (AA), collagen, and ADP in rabbit platelet-rich plasma.

Compound^b	IC50 values^a (µM) or inhibition (%) at 100 µM		Inhibition (%) at 100 µM
	Arachidonic acid (200 µM)	Collagen (5 µg/ml)	ADP (5 µM)
Indomethacin	0.6^c	3.1^c	—
9a	30.1 (29.8–30.4)	156.1 (114.4–212.9)	14.1
9b	0.0%	3.2%	15.2
9c	7.9 (6.2–10.2)	47.7 (39.7–57.3)	15.0
9d	1.2 (0.4–3.4)	8.2 (6.0–11.1)	2.2
9e	0.7 (0.6–1.1)	4.5 (3.9–5.1)	25.7*
9f	2.0 (1.9–2.2)	6.0 (4.9–7.4)	18.7
9g	47.4 (43.8–51.3)	17.7%	15.6
9h	11.4%	2.1%	18.3
9i	1.7 (0.9–3.1)	6.1 (4.3–8.5)	1.4
9j	23.2 (23.1–23.4)	41.2 (32.7–51.7)	22.6*
LASSBio-294	15.3^c	18.3^c	14.7^c

^an = Three independent experiments in duplicate; 95% confidence interval for IC₅₀ values is shown in parentheses. ^bN-acylhydrazone derivatives 9a–j were incubated with platelet-rich plasma 5 min before the addition of the agonist (AA, collagen, or ADP). ^cData obtained from reference⁸. *P < 0.05 (ANOVA one-way; Dunnet posttest).

Figure 3.  McLafferty rearrangement peak in the mass spectra of (E)-(2-furfurylidene) 3,4-methylenedioxybenzoylhydrazine derivative 9a.

Table 2.  Effect of ASA and functionalized 2-furyl N-acylhydrazone derivatives 9a, 9c–e, and 9i–j on platelet aggregation induced by ADP (3 µM) in human platelet-rich plasma.

Compound^a	n^b	Aggregation^c (%)	Total inhibition (%)	Inhibition (%)
				First wave	Second wave
ADP	4	64.0 ± 4.4	—	—	—
DMSO	4	69.3 ± 7.8	—	—	—
ASA	3	53.0 ± 5.5	23.5	−4.0 n.s.	96.5*
9a	4	58.0 ± 6.0	16.2	−0.5 n.s.	41.7*
9c	4	48.5 ± 3.0	30.0*	0.5 n.s.	88.3*
9d	4	52.5 ± 6.0	24.5	−2.9 n.s.	96.1*
9e	4	42.5 ± 3.8	38.6*	9.7 n.s.	97.4*
9f	4	55.8 ± 6.6	19.5	−8.8 n.s.	94.7*
9i	4	55.5 ± 3.8	19.9	−4.4 n.s.	97.0*
9j	4	69.0 ± 7.6	0.4	−7.9 n.s.	22.4
LASSBio-294	—	—	40.9^d	—	—

^aCompounds were assayed at 100 µM concentration. ^bn = number of independent experiments in duplicate. ^cResults are expressed as mean ± SEM. ^dData obtained from reference⁸. N-acylhydrazone derivatives 9 were incubated with plasma-rich platelet 5 min before the addition of the ADP. *P < 0.05 (ANOVA one-way; Dunnet posttest).

Figure 4.  Effect of NAH derivatives 9a, 9e, and 9j on the TXB₂ production in arachidonic acid (500 µM)-induced platelet aggregation in human PRP. Results are expressed in terms of mean ± SEM (n = 3–5 independent experiments). *P < 0.05 (ANOVA one-way; Dunnet posttest); N-acylhydrazone derivatives 9 were incubated with plasma-rich platelet 5 min before the addition of AA; The vehicle concentration in the samples does not exceed 0.3%; INDO = indomethacin.

Brito FC, Kummerle AE, Lugnier C, Fraga CA, Barreiro EJ, Miranda AL. Novel thienylacylhydrazone derivatives inhibit platelet aggregation through cyclic nucleotides modulation and thromboxane A2 synthesis inhibition. Eur J Pharmacol 2010;638:5–12.

 PubMed Web of Science ®Google Scholar