Main chemical constituents and pharmacological properties of Harrisonia perforata (Blanco) Merr.

Shifng Li, Zhiyng Yn, Xioxio Hung＊, Shojing Song＊

a Department of Natural Product Chemistry, Shenyang Pharmaceutical University, Shenyang 110016, China;

b Chinese People's Liberation Army 210 Hospital, Dalian 116021, People’s Republic of China

1 Introduction

Harrisonia perforata (Blanco) Merr., the only species of Harrisonia genus, belongs to the Simarubaceae family. It is mainly distributed in southeast of Asia and southern China [1]. Its roots and leaves are applied for the treatment of wound healing and malaria in Chinese folk medicine [2].In Vietnamese folk medicine, it is used to treat itching, and also used for diarrhea and dysentery in neighboring countries [3]. Extensive studies indicated that the main chemical constituents of this plant are structurally diverse chromones, rearranged limonoids, polyketides and triterpenes [4].According to the previous studies of H. perforata,it exhibited various pharmacological activities, such as anti-cancer, anti-inflammatory, antibacterial and insecticidal activities [5-8]. This paper summarized the chemical constituents and pharmacological activities of H. perforata, which provides a foundation for the further researches.

2 Studies of chemical constitutions

There have been some reports about the chemical compositions of H. perforata. And it is found that the main compounds of H. perforata are limonoids, chromones according to the previous researches.

2.1 Limonoids

There were about twenty-eight limonoids isolated from H. perforata (Fig. 1). Three compounds were isolated from the roots of this plant: harperfolide (1), harrisonin (2), obacunone(3) [6]. From the leaves and branches of H.perforata, kihadanin A (4), kihadanin B (5),6α-acetoxyobacunol acetate (6) [9], haperforine A (7), 12-desacetylhaperforine A (8), haperforine E (9) [3], perforanoid A (10) [10], perforatinolone(11) [11], perforatin (12) [12], haperforin B1(13), haperforin D (14) [13], haperforin C2(15), haperforin F (16), haperforin G (17) [14],harperspinoid A (18), harperspinoid B (19) [2] were isolated. Perforin A (20), 12β-acetoxyharrisonin(21), harrpernoid B (22), harrpernoid C (23),5,6-dehydrodesepoxyharperforin C2 (24), rutaevine(25) [15], haperforatin (26) [6] were extracted from fruits of H. perforata. Two limonoids were isolated from the twigs and stems of H. perforata:perforalactone A (27), perforalactone B (28) [8].

Fig. 1 Limonoids in Harrisonia perforata

2.2 Prenylated polyketides

Five compounds possessing an unusual spirocyclic skeleton (Fig. 2), harrisotone A(29), harrisotone B (30), harrisotone C (31),harrisotone D (32), harrisotone E (33) and a new hydroperoxypolyketide harrisonol A (34) [5] were isolated from the stems and leaves of H. perforata.

Fig. 2 Prenylated polyketides in Harrisonia perforata

2.3 Chromones

Previous studies have showed that the isolated chromones from H. perforata can be mainly divided into two classes: chromones and methyl chromones derivatives (Fig. 3). Chromones included heteropeucenin-5-methoxy-7-methyl ether (35),perforatin B (36) [16], perforatin D (37), perforatin E (38), perforatin F (39), heteropeucenin-5-methoxy-7-methyl ether (40), heteropeucenin-7-methyl ether(41) [17], heteropeucenin-7-methyl ether (42) [16],(±)-perforatin C (43a, 43b), (±)-2'-O-acetylperforatin C (44a, 44b), (±)-erythro-1'-hydroxyperforatin C (45a, 45b) [18], peucenin-7-methyl ether(46), perforamone A (47), perforamone B (48),perforamone C (49), eugenin (50), saikochromone A (51) [7], 5-hydroxy-7-methoxy-2-methyl-8-[(1E)-3-oxo-1-butenyl]chromone (52) [18], harriperfin E (53) [9]. And greveiglycol (54), greveiglycol 4'-methyl ether (55), (±)-horriperfin A (56a, 56b),(±)-horriperfin B (57a, 57b) [18], perforatic acid methyl ester (58), perforatic acid (59) [18], perforatin A (60) [16], 2-hydroxymethylallopataeroxylin-5-methyl ether (61), perforatic acid methyl ester(62), perforamone D (63) [17], umtatin (64) [15],greveichromenol (65) [7], perforatins G (66) [17]belong to methyl chromones derivatives.

Fig. 3 Chromones in Harrisonia perforata

2.4 Other compounds

Other compounds mainly included gardaubryones C (67) [9], pachymic acid (68) [15],stigmasterol (69), β-sitosterol (70) [1], β-sitosterol methyl ether (71) [9], pinoresinol (72) [15],caryolane-1, 9β-diol (73), β-daucosterol (74) [1],palmitic acid (75), linoleic acid (76) [23] (Fig. 4).

Fig. 4 Others in Harrisonia perforata

3 Studies of pharmacological activities

3.1 Anti-cancer activity

Yin [5] et al. isolated five novel prenylated polyketides, harrisotones A-E and a hydroperoxypolyketide harrisonol A from the stems and leaves of H. perforata. Harrisotones A-C (29-31) and the hydroperoxypolyketide harrisonol A (34) were evaluated for the cytotoxic activities against P-388 and A-549 tumor cell lines, using MTT and SRB methods. The results showed that harrisotone A, C and harrisonol A had significant cytotoxic activities against P-338 cell line, and the IC50values were 1.56,2.35 and 0.27 μM, respectively. Harrisotone A and harrisonol A also showed moderate activities against A-549 tumor cell line with IC50of 24.5 and 26.6 μM,respectively, as pseudolaric acid B was used to be the positive control with IC50values of 0.74 and 1.99 against P-338 and A-549, respectively.

Eight limonoids, isolated from the fruits of H. perforata by Yan [15] et al., were tested the cytotoxicities against the human leukaemia HL60 and human lung adenocarcinoma A-549 cell lines in vitro. Among these compounds, harrpernoid B (22)showed weak activity against HL60 and A-549 cell lines with the inhibition rates of 63.6% and 64.9%,respectively, at the concentration of 10 μM.

It was reported that perforatic acid (59) and heteropeucenin-7-methyl ether (41) extracted from H. perforata could inhibit effectively3H-TdR from permeating hepatoma ascites cells of mouse [19,20].

3.2 Anti-malarial activity

Nguyen-Pouplin [21] et al. investigated traditional medicinal plants used to treat malaria and identified forty-nine plants. 228 extracts from these plants were tested for their activities against Plasmodium falciparum in vitro. The results illuminated that the extracts of H. perforata had inhibitory effect on Plasmodium falciparum with IC50value of 5.1 μg/ml. Perforatin A (60)was isolated from the branches of H. perforata by Tuntiwachwuttikul [7] et al. and discovered that it had weak anti-plasmodial activity against Plasmodium falciparum, and the EC50value was 10.5 μg/ml.

3.3 Anti-inflammatory activity

Choodej [6] et al. isolated two new limonoids and a new chromone together with eight known compounds from fruits and roots of H. perforata.These compounds were evaluated for their antiinflammatory activity by testing the inhibition of NO production in LPS-activated murine macrophage J774. A1 cells. The results revealed that harperfolide(1) expressed the strong activity, with IC50value of 6.51 ± 2.10 μM, compared to control group with IC50of 28.42 ± 3.51 μM. Furthermore, the toxicity of harperfolide was investigated on unstimulated cell lines by measuring the cell viability. The outcome indicated that harperfolide did not showed notable cytotoxicity on macrophage J774. A1 cells. Thus,harperfolide inhibited NO production without causing cell death.

3.4 Antibacterial activity

Four new chromones together with six known compounds were isolated from the branches of H. perforata by Tuntiwachwuttikul [7] et al. The compounds were assessed for antimycobacterial activities against Mycobacterium tuberculosis H37Ra, using isoniazide (MIC of 0.040-0.090 μg/ml)and kanamycin sulfate (MIC of 2.0-5.5 μg/ml)as reference compounds. The results showed that perforamone B, D (48, 63) demonstrated weak antimycobacterial activities, with MIC of 25 μg/ml and 25 μg/ml, respectively.

Limsong [22] et al. studied the inhibiting action on adherence of Streptococcus mutans (S. mutans)ATCC 25175 and TPF-1 with the ethanol extracts of six herbs in vitro. In glass surface adherence assay,H. perforata showed the strongest inhibition for S. mutans ATCC 25175 with adherence of 4.7% to control, and was the strong inhibitor for S. mutans TPF-1 with adherence of 7.6% to control at 0.5%concentration. Subsequently, in bacterial adherence to hydroxyapatite assay, H. perforata expressed less than 50% adherence on S. mutans ATCC 25175.These findings indicated that H. perforata could inhibit the adherence of S. mutans in vitro, and may be effective at controlling dental caries.

3.5 Insecticidal activity

Fang [8] et al. isolated three quassinoids from H. perforata, and perforalactone A, B (27,28) were tested for insecticidal activity against Aphis medicaginis Koch. The result showed that perforalactone B had outstanding activity against Aphis medicaginis Koch, and the LC50values was 7.23 μM, but perforalactone A almost had no activity.In addition, the mechanism of their insecticidal effect was studied with nicotinic acetylcholine receptor (nAChR), a major and ubiquitous target for insecticide action. It showed that perforalactone B was more active than perforalactone A, with the IC50of 1.26 nM and 15.8 nM, respectively.Specially, the IC50of perforalactone B was similar to that of imidacloprid, the most commercially important insecticide. Besides, biochemical electrophysiological tests indicated that they were all antagonists at the nAChR of the insect.

4 Conclusion

Currently, the studies about H. perforata have been developed at home and abroad, but there are not many researches on its chemical components and pharmacological activities. We summarized the chemical constituents and pharmacological properties of H. perforata according to the previous reports, and the effective compounds to treat cancer,malaria, inflammation, etc. were discovered, which is meaningful for enriching the medicinal resources.This paper will be beneficial to the subsequent researches about H. perforata.

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