Cachrys L. Genus: A Comprehensive Review on Botany, Phytochemistry and Biological Properties

The Cachrys L. genus belongs to the Apiaceae family and it is widely distributed in the Mediterranean basin, with plant species being endemic to southern Europe, Asia, and northern Africa. Different studies, focused on the phytochemical composition of Cachrys spp. and the biological properties of their phytocomplexes, have been reported. These works mostly focused on the essential oils obtained from these plants, and pointed out that Cachrys species are a rich source of coumarins, mainly furanocoumarins. Other phytochemicals, such as terpenes, fatty acids, phytosterols, and flavonoids have been also identified. Moreover, a number of biological properties such as antioxidant, antimicrobial, anti-inflammatory, cytotoxic, and photocytotoxic effects have been assessed. Nevertheless, a review of the chemical and pharmacological properties of this genus is not available in the literature. The aim of this paper is to provide an overview of the reports concerning the identified phytochemicals and the biological effects reported for Cachrys spp., and to offer a comprehensive understanding of the potential of this genus as a source of bioactive compounds. The current taxonomy, the traditional uses, and the toxicological aspects of plants belonging to this genus are also reported, and the future research directions are discussed.

presents a bubble chart showing the number of papers dealing with the different Cachrys species, and the period of publication. As shown, the interest towards these species is increasing in the last years, with C. microcarpos M.Bieb., C. sicula L., C. cristata DC., and C. libanotis L. being the most investigated plants. Figure 3 presents a bubble chart showing the number of papers dealing with the different Cachrys species, and the period of publication. As shown, the interest towards these species is increasing in the last years, with C. microcarpos M.Bieb., C. sicula L., C. cristata DC., and C. libanotis L. being the most investigated plants.

Taxonomy of the Cachrys Genus
The classification of the Cachrys genus has been somewhat confused. According to Tutin (1968), this genus included also Prangos Lindley and Hippomarathrum Link, while in 1872, Boissier had classified them into three different genera [15][16][17].
Apiaceae is a large family with a complex taxonomy, which comprises 430 genera and about 3780 species, most of which belong to the subfamily Apioideae [18].
According to the molecular phylogenetic studies by Mousavi and colleagues, the genera Azilia, Bilacunaria Pimenov & V.N.Tikhom., Cachrys L., Diplotaenia Boiss., Eriocycla, Ferulago W.D.J.Koch, and Prangos belong to the Cachrys clade, and are all widely distributed from southern Europe, northern Africa, Turkey, central and Southwest Asia, to China [18]. Table 1 reports the included species and the taxonomic synonyms.

Taxonomy of the Cachrys Genus
The classification of the Cachrys genus has been somewhat confused. According to Tutin (1968), this genus included also Prangos Lindley and Hippomarathrum Link, while in 1872, Boissier had classified them into three different genera [15][16][17].
Apiaceae is a large family with a complex taxonomy, which comprises 430 genera and about 3780 species, most of which belong to the subfamily Apioideae [18].
According to the molecular phylogenetic studies by Mousavi and colleagues, the genera Azilia, Bilacunaria Pimenov & V.N.Tikhom., Cachrys L., Diplotaenia Boiss., Eriocycla, Ferulago W.D.J.Koch, and Prangos belong to the Cachrys clade, and are all widely distributed from southern Europe, northern Africa, Turkey, central and Southwest Asia, to China [18]. Table 1 reports the included species and the taxonomic synonyms.

Botanical Description and Ethnobotanical Uses
Cachrys species are described as much-branched, robust perennial herbs with deeply dissected leaves. They have compound umbels and hermaphrodite flowers. The calyx has five short and distinct teeth, and petals are obovate, usually yellow. The fruit is ovoid to subglobose, crowned with a broad disk [1,19].
Unlike other genera, whose plants have been widely empirically used in popular medicine for their beneficial effects, there is not much information in the literature about the traditional uses of Cachrys species.
Cachrys libanotis has been reported to be traditionally used in the treatment of rheumatism [20]. Cachrys alpine M.Bieb. is instead used as an aphrodisiac in Balikesir province in Western Turkey, where it is known as 'Çaksir otu' [21].
In Eastern Anatolia (Turkey) many wild edible plants are traditionally used in daily diets. These plant species are commonly used as fresh vegetables, dried herbs, or as pickled plants in some sole or mixed milk and meat products [22]. C. cristata DC. (Hippomarathrum cristatum Boiss.) is also traditionally used in Turkey, where it is known as "Tarhana otu" or "Çaşır", and is added to a traditional tarhana soup [23].
Moreover, in the Eastern Anatolia region of Turkey, C. microcarpos M.Bieb. is still used by the farmers as an alternative source of roughage. Even if there are few studies in the literature concerning the use of the H. microcarpum plant as a source of coarse feed in animal feeding, the nutritional content of this plant has been reported by Gül [24].

Phytochemical Constituents
Phytochemical studies on the Cachrys species allowed the characterization of coumarins, terpenoids, phenylpropanoids, phenolic compounds, fatty acids, and phytosterols. Overall, the phytochemical investigations carried out till now allowed the identification of 177 secondary metabolites. The remarkable furanocoumarin content which characterizes Cachrys species is particularly interesting, given the growing importance that these compounds are assuming in dermatology and oncology. Indeed, furanocoumarins are useful photosensitizing agents in photodynamic therapy. Most of the studies concerned the phytochemical characterization of essential oils from Cachrys species, but in some cases, extracts obtained through maceration or Naviglio ® extractor have also been analyzed.

Coumarins
Coumarins, mainly furanocoumarins, stand out among the main phytochemical compounds identified in Cachrys species.
Coumarins (1,2-benzopyrone or 2H-1-benzopyran-2-one) and their derivatives are natural compounds widely found both as a heteroside and free form in the seeds, roots, and leaves of many plant species, mainly belonging to the Apiaceae and Rutaceae families [25,26]. Furanocoumarins are tricyclic aromatic compounds bearing a furan ring fused to the α-benzopyrone (coumarin) system. These derivatives can be structurally divided into angular and linear furanocoumarins, with the furan ring attached at the 6,7 positions on the aromatic ring (e.g., angelicin) or to the 7,8 positions (e.g., psoralen), respectively [27].
Marrelli and colleagues investigated the phytochemical profile of Cachrys sicula and Cachrys libanotis extracts obtained through two different extraction techniques, a traditional solvent maceration and a pressurized cyclic solid-liquid extraction technique, the Naviglio extraction ® . The GC-MS qualitative profile revealed the presence of three furanocoumarins: xanthotoxin, bergapten and isopimpinellin, in each investigated extract. Xanthotoxin was identified in the highest amount in both the extracts obtained with Naviglio ® , with percentages equal to 14.8% and 10.8% for C. libanotis and C. sicula, respectively. Bergapten, another naturally occurring furanocoumarin, was detected with percentages from 1.8% to 2.8%. The same trend was observed as regards isopimpinellin. A quantitative analysis was also carried out. Consistent with previous analyzed data, Cachrys libanotis and Cachrys sicula extracts, both obtained through the innovative Naviglio extraction ® technique, showed the highest amount of xanthotoxin, with values equal to 4.98 ± 0.21 and 4.10 ± 0.23 mg/mL, respectively. As regards bergapten, the same trend can be observed, Cachrys libanotis extract obtained with the Naviglio ® extractor being the richest sample both in bergapten and xanthotoxin. On the contrary, isopimpinellin showed the highest value in the Cachrys sicula extract obtained through the Naviglio ® instrument. Seselin (a pyranocoumarin), as well as osthol, suberosin, and isogeijerin (coumarins), were found in Cachrys libanotis macerated extract, while the pyranocoumarin 2-methyl-2-butenoic acid 9,10-dihydro-8,8-dimethyl-2-oxo-2H,8Hbenzo[1,2-b:3,4-b0] dipyran-9-yl ester was recognized in Cachrys sicula and Cachrys libanotis extracts obtained through the Naviglio ® extractor at 10.2% and 9.7%, respectively [5].
De Leo and colleagues isolated for the first time the compound 8-hydroxymethylpso ralen, a new coumarin, together with the known sprengelianin and (RS)-oxypeucedanin in Cachrys sicula leaves extract [30].

Terpenoids
A huge number of terpenoids have been identified in species belonging to Cachrys genus. Due to the high content of this class of phytochemicals, a Cachrys species, namely C. cristata DC., was one of the 21 plant species from Greece investigated by Evergetis and coworkers for their potencies as essential oils (EOs) producing crops and as renewable sources for the production of fine chemicals. C. cristata resulted one of the most potent EO producing crops with an estimated production yields per hectare exceeding the 30 L, with the compound myrcene accounting for estimated 19 L [31].
Baser and coworkers described the phytochemical composition of the essential oil from the fruit of C. alpine M.Bieb from Turkey [21]. The authors reported that the microdistilled oil from the dried fruits of this species contained different terpenes, with α-humulene being the major constituent (33.1%). p-Cymene, α-phellandrene, and germacrene D were present in percentages above 8%, and α-pinene (6.3%) was also identified among the most abundant components. β-Pinene, β-phellandrene, β-caryophyllene, bicyclogermacrene, germacrene B, and other minor constituents were also identified.
The phytochemical composition of a C. sicula essential oil was also reported by Tahar and coworkers, who analyzed the essential oil from the leaves of the plant. Thirty-two compounds, accounting for about 99% of the total oil, were identified, with β-pinene, sabinene, myrcene, and α-pinene being the main constituents [32].
The phytochemical profile of Cachrys sicula and Cachrys libanotis extracts, obtained through both maceration and Naviglio ® extractor, showed the ubiquitous presence of the terpene estragole, while limonene, fenchone, and anethole were only detected in the Cachrys sicula extract obtained with the Naviglio ® extractor [5].
The composition of the essential oil from this same species was also investigated by Khalilzadeh and colleagues, who assessed the phytochemical content of the essential oil from the aerial parts of C. microcarpos M.Bieb. (syn. Hippomarathrum microcarpum) obtained by hydrodistillation. The authors reported α-pinene, β-caryophyllene, β-phellandrene, and germacrene-D as the major components [8].
Furthermore, the chemical composition of C. microcarpos was also investigated by Sefidkon and Shaabani, who described the phytochemical content of the essential oil from the leaves and flowers of the plant. β-Caryophyllene, y-muurolene, and linalool were found to be the major components of the leaf essential oil. These same compounds, together with thymol and camphene, were also the main constituents of the essential oil obtained from the flowers [36].
Özek and coworkers verified the phytochemical composition of the essential oil from the fruits of this same Cachrys species. The plant material from C. cristata was subjected to hydrodistillation, and the analysis were carried out with GC-MS, with sesquiterpenes resulting as the most abundant class of terpenes (accounting for about 38% of the oil), and germacrene D and germacrene B being the major identified terpenoids [23].
The phytochemical composition of C. boissieri (Reut. & Hausskn. ex Boiss.) Hand (syn. Hippomarathrum boissieri) was also investigated. Baser and coworkers reported the terpenoids identified in the essential oil from the aerial parts of this species, which were distilled using a Clevenger-type apparatus. GC-MS analyses revealed the presence of eighty-nine compounds, with β-caryophyllene, caryophyllene oxide, and α-pinene (8.8%) being the major compounds [37].

Phenylpropanoids
The presence of some phenylpropanoids (4.9%) was reported for C. cristata. Myristicin was found to be the major component belonging to this class of compounds in the essential oil obtained from the fruit of this species, followed by methyl eugenol [23].
The phenylpropanoid eugenol has been also reported in the essential oil from the aerial parts of C. boissieri [37].

Phenolic Compounds
Few data on the phenolic composition of Cachrys spp. have been reported. The ethyl acetate fraction of C. pungens aerial parts in methanolic extract was explored using highperformance thin-layer chromatography (HPTLC), which allowed identification of the flavonoid catechin, the flavonoid glycosides naringin and quercitrin, the cinnamic acids caffeic and ferulic acids, and also gallic acid, a phenolic acid [38].

Fatty Acids
The presence of some fatty acids has been reported for C. sicula and C. libanotis. Palmitic acid was identified in both Cachrys species, while myristic acid and α-linoleic acid were identified in percentages equal to 0.2% and 0.7% in Cachrys libanotis extracts obtained through Naviglio ® extraction and traditional solvent maceration, respectively [5].
The n-hexane fraction of C. pungens extract obtained through maceration revealed the presence of hexadecanoic acid (palmitic acid) and 8,11-octadecadienoic acid methyl ester in percentages equal to 2.5% and 2.2%, respectively, together with some other minor constituents belonging to this class, among which was linoleic acid (9,12-octadecadienoic acid) [6].
Küçükboyacı and coworkers carried out an interesting study aimed to assess the fatty acid composition of the seed oil from a number of Apiaceae species from Turkey, including C. cristata DC. (Hippomarathrum cristatum Boiss.) and C. microcarpos (Hippomarathrum microcarpum). In particular, the purpose of the authors was to verify the content of petroselinic acid, a valuable oleochemical raw material whose double bond can be oxidatively split to form lauric and adipic acids, two fatty acid used for the production of surfactants and soaps. Petroselinic acid was found to be the major fatty acid present in the seed oils of C. cristata (72.2%). Linoleic acid was instead the most abundant component of C. microcarpos seed oil (36.9%) [13].
Özek and coworkers verified the phytochemical composition of the essential oil from the fruits of C. cristata, reporting the presence of hexadecanoic acid (palmitic acid), accounting for 11.6% of the essential oil, together with other fatty acid methyl and ethyl esters [23].
Moreover, Pinar and Alemany reported the isolation of N-N'-di-o-tolylethylendiamine from C. sicula. This molecule was identified in an acidic fraction of the methanolic extract, and it was the first time this compound had been isolated from a natural source [39]. Table 2. summarizes the data focused on the phytochemical characterization of plants belonging to the Cachrys genus.

Biological Properties of Cachrys Species
The analysis of the literature allowed us to highlight some interesting biological properties for plants belonging to the Cachrys genus, namely antioxidant, antibacterial, antifungal, cytotoxic, and photocytotoxic potential, together with interesting enzyme inhibitory properties and even insecticidal activity (Figure 4a). However, this genus still appears to be extremely understudied. The number of papers dealing with its biological activity is limited. We find just seven papers focusing on the antioxidant effects of Cachrys species and five reports about the antimicrobial potential. The number of papers dealing with the remaining activities is even lower. Overall, C. libanotis appears to be the most investigated species with regards to the biological potential, followed by C. microcarpos and C. sicula, for which four biological activities have been reported to date (Figure 4b).
appears to be extremely understudied. The number of papers dealing with its biological activity is limited. We find just seven papers focusing on the antioxidant effects of Cachrys species and five reports about the antimicrobial potential. The number of papers dealing with the remaining activities is even lower. Overall, C. libanotis appears to be the most investigated species with regards to the biological potential, followed by C. microcarpos and C. sicula, for which four biological activities have been reported to date (Figure 4b).

Antioxidant Activity
Aouachria and coworkers explored the antioxidant potential of the hydroalcoholic root extract of C. libanotis, which was assessed on superoxide generated by an NADH/PMS system, DPPH test, FRAP, ferrous iron chelation, and β-carotene bleaching assays. The crude extract and its aqueous fraction showed a good chelating activity, with IC 50 values equal to 52.6 ± 0.3 and 52.7 ± 0.2 µg/mL, respectively. The crude extract also showed the highest radical scavenging activity (IC 50 = 0.41 ± 0.01 mg/mL). Moreover, all the fractions were effective in protecting linoleic acid from peroxidation [20].
Marrelli and colleagues investigated the in vitro potential antioxidant activity of two Cachrys species from southern Italy, C. sicula and C. libanotis, extracted through maceration and also using the Naviglio ® extractor, an innovative solid-liquid extraction technique. The macerated extracts showed the best radical scavenging activities, with IC 50 values of 102.13 ± 0.79 and 112.73 ± 0.88 µg/mL for C. libanotis and C. sicula, respectively, in the DPPH assay. The inhibition of lipid membrane peroxidation, investigated through β-carotene bleaching test, confirmed the trend observed for free radical scavenging activity, demonstrating that the macerated extracts significantly protected linoleic acid after 30 min of incubation (IC 50 values equal to 16.77 ± 1.43 and 19.22 ± 1.07 µg/mL for C. sicula and C. libanotis extracts, respectively) [5].
Tahar and coworkers assessed instead the antioxidant potential of the essential oil from the leaves of C. sicula, using the well-established ABTS, DPPH, and metal chelating assays. A modest DPPH radical scavenging was observed, with an IC 50 value equal to 267.94 ± 0.00 µg/mL, while the essential oil demonstrated a better antioxidant activity in the ABTS test (IC 50 = 81.93 ± 0.00 µg/mL) [32].
Menichini and colleagues evaluated the antioxidant activity of C. pungens alcoholic extract and fractions. The ethyl acetate fraction showed the best radical scavenging activity, with IC 50 equal to 12.15 ± 0.02 µg/mL. Consistently, the same fraction as well as the total extract showed the best linoleic acid protective activity in the β-carotene bleaching assay, with IC 50 values equal to 8.33 ± 0.33 and 9.16 ± 0.27 µg/mL after 30 min, respectively. The same trend was observed after 60 min of incubation [6].
Extracts prepared starting from the aerial parts of two C. crassiloba (Boiss.) Meikle accessions collected in Turkey were investigated for their antioxidant potential through the DPPH assay. The extracts neutralized the DPPH radical, with percentages of inhibition equal to 67.611% and 81.281%, respectively [40].
Matejic and coworkers investigated the antioxidant potential of the aerial parts and fruits of C. cristata extracts by means of both the DPPH and ABTS assays. The fruit water extract showed the best radical scavenging activity (IC 50 = 1.784 mg/mL), followed by the methanolic extracts of the fruits and aerial parts (IC 50 values of 3.347 and 4.058 mg/mL, respectively). As regards the ABTS test, the acetone extract of C. cristata aerial parts showed the highest antioxidant potential, with a value of 3 mg of vitamin C per mL of extract [41].
C. microcarpos (synonym Hippomarathrum microcarpum) essential oil and methanolic extract were evaluated for their antioxidant activity by means of the DPPH and linoleic acid assays. The extract did not show any radical scavenging activity, while the essential oil exhibited an IC 50 value of 10.690.0 ± 0.5 µg/mL (DPPH assay) [34].

Antimicrobial Activity
The antibacterial activity of C. libanotis was tested on different bacterial strains by Aouachria and colleagues. The hydroalcoholic root crude extract was effective against Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus cereus, while its chloroform and ethyl acetate fractions showed an antibacterial effect against Bacillus cereus, Enterococcus faecalis, and Lysteria monocytogenes [20].
Matejic and colleagues investigated the antimicrobial activity of C. cristata extracts against different strains of Gram-positive and Gram-negative bacteria. The ethyl acetate extract of C. cristata aerial parts showed both an inhibitory and a bactericidal activity against Gram-positive bacteria in concentrations ranging from 0.78 to 6.25 mg/mL, while inhibitory effects of the same extract were assessed against Pseudomonas aeruginosa without exerting any bactericidal activity at the highest tested dose [41].
Cachrys cristata leaf essential oil exerted antibacterial activity against Staphylococcus aureus, Micrococcus luteus, and Staphylococcus epidermidis bacteria. The stem essential oil produced by hydrodistillation starting from the same species was effective against Staphylococcus aureus Gram-positive bacteria, and showed a quite good activity against Candida albicans [33].
Ozer and colleagues investigated the antimicrobial and antifungal properties of C. microcarpos essential oil and methanolic extract. The samples exerted antimicrobial activity against eight and two bacterial strains, respectively. As the antifungal activity was also assessed, the essential oil showed the best activity by inhibiting one yeast (Candida albicans) and nine fungal strains [34].
Karakaya and colleagues also tested the essential oil from C. microcarpos aerial parts for its antimicrobial activity, reporting efficacy against Candida albicans and Staphylococcus aureus strains [35].

Anti-Inflammatory Potential
Perri and coworkers (2022) investigated the in vitro potential anti-inflammatory properties of two different Cachrys species collected in southern Italy and extracted through the Naviglio ® instrument. C. libanotis and C. pungens extracts were tested for the inhibition of pro-inflammatory cytokines (TNF-α and IL-6) and NO mediator in RAW 264.7 cells activated with LPS. The data showed that both the extracts significantly inhibited the release of TNF-α and IL-6, and the production of NO. Moreover, C. pungens induced the release of the anti-inflammatory cytokine IL-10. Extracts were also tested for their ability to inhibit the JAK/STAT signaling pathway: both the tested samples inhibited the phosphorylation of JAK2 and STAT3 proteins, with the C. pungens extract being the most effective one [9].
C. microcarpos (synonym Hippomarathrum microcarpum) ethanolic extract demonstrated anti-inflammatory effects in carrageenan-induced oedema in rats, showing, at high doses, good anti-inflammatory activity and decreasing inflammation rates better than diclofenac [42].

Antiproliferative Activity
The antiproliferative effects of Hippomarathrum microcarpum (synonym Cachrys microcarpos M.Bieb.) aerial parts extract were tested on human prostate cancer cells (PC-3) by using the MTT assay. Both the ethanolic and the aqueous extracts inhibited cell viability after 24 h, with IC 50 values of 12.99 and 23.34 mg/mL, respectively [43].
De Leo and colleagues tested for their antiproliferative activity three pure coumarins they isolated from the leaves of C. sicula L.: the new coumarin 8-hydroxymethylpsoralen and the known compounds sprengelianin and oxypeucedanin. The cytotoxic effects were assessed on Jurkat and HeLa cancer cell lines. However, 8-hydroxymethylpsoralen was the only compound demonstrating a moderate inhibitory activity [30].

Photocytotoxic Activity
The photocytotoxic activity observed for Cachrys species is linked to their furanocoumarins content [44].
Growing evidence underlines the anticancer potential of furanocoumarins. The general mechanism by which these compounds exert their effects on cancer cells is based on cell cycle blockage and programmed death, like apoptosis or autophagy [45]. Moreover, experimental studies have shown that furanocoumarins activate multiple signaling pathways, also leading to antioxidant and antimetastatic effects, and cell cycle arrest in malignant cells. Furanocoumarins have also been demonstrated to show synergistic potential with anticancer drugs [46].
Beside their antiproliferative effects, furanocoumarins are very interesting molecules since they are useful in photochemotherapy, a promising therapeutic approach in anticancer research which combines the action of a light source and a chemical photosensitizer [26,47].
PUVA therapy (Psoralen + UVA therapy) is based on the administration of psoralens followed by exposure to UVA radiation [44]. Beside its use in the treatment of dermatological diseases such as psoriasis and vitiligo, it is used in the treatment of cutaneous T-cell lymphoma [48][49][50][51].
Menichini and colleagues investigated the photocytotoxicity of C. pungens methanolic extract and its fractions on A375 melanoma cells irradiated at 365 nm at a dose of 1.08 J/cm 2 . The chloroform and coumarinic fractions exerted the best photocytotoxic activity, with IC 50 values equal to 0.286 ± 0.067 and 0.209 ± 0.033 µg/mL, respectively, while they did not affect the viability of nonirradiated cells [6].
C. sicula and C. libanotis extracts photocytotoxic effects were evaluated on C32 human melanoma cells irradiated with UV light (365 nm) for 1 h at a dose of 1.08 J/cm 2 . Both the aerial parts methanolic extracts obtained throug maceration and the Naviglio ® extractor were evaluated. The extracts obtained using the Naviglio ® extractor significantly inhibited cell viability, with IC 50 values of 3.16 ± 0.21 and 8.83 ± 0.20 µg/mL for C. libanotis and C. sicula, respectively. These two samples were also demonstrated to induce up-regulation of apoptotic signals such as Bcl2-associated X protein (BAX), and poly ADP-ribose polymerase (PARP) cleavage. Moreover, the apoptotic responses were also verified, and these samples proved to be more photoactive, causing a greater upregulation of the p21 protein in the presence of UVA radiation compared to the extracts obtained through traditional maceration [5].

Acetylcholinesterase (AChE) and Butyrylcholinesterase (BChE) Inhibitory Activity
Acetylcholinesterase (AChE) is the key enzyme in the breakdown of acetylcholine, and its inhibition is a target for the treatment of Alzheimer's disease and other neuro-logical disorders including senile dementia, myasthenia gravis, and ataxia [52]. Butyrylcholinesterase (BuChE) plays a minor role in regulating brain acetylcholine levels, but the activity of this enzyme increases in patients with Alzheimer's disease (AD) [53]. Different plant extracts have been demonstrated to exert interesting inhibitory effects on these enzymes [52,54]. Tahar and coworkers evaluated the inhibitory properties of C. sicula on the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes. The essential oil was obtained from the hydrodistillation of the leaf of the plant, and the biological activity was measured using acetylthiocholine iodide and butyrylthiocholine chloride as substrates of the reaction. C. sicula essential oil exerted a good inhibitory activity on BChE, with an IC 50 value equal to 91.90 ± 0.00 µg/mL, and a mild inhibitory activity on AChE (IC 50 = 169.91 ± 0.00 µg/mL) [32].

Xanthine Oxidoreductase (XOR) Inhibitory Activity
The xanthine oxidoreductase (XOR) enzyme catalyzes the oxidative hydroxylation of hypoxanthine to xanthine and xanthine to uric acid and it is considered a target for the treatment of hyperuricemia and gout [55].
Aouachria and coworkers demonstrated a good xanthine oxidoreductase (XOR) inhibitory effect for C. libanotis root extract. To perform the experiments, the enzyme was purified from bovine milk and the inhibitory activity was measured using a spectrophotometric method, measuring uric acid production at 295 nm. Allopurinol was used as a positive control. The roots of the plants were powdered with a traditional mill and extracted with a hydroalcoholic solution. The raw extract was then fractionated using solvents with increasing polarity with liquid-liquid extraction, to achieve the corresponding hexane, chloroform, and ethyl acetate fractions. A good xanthine oxidoreductase (XOR) inhibitory effect was observed for the ethyl acetate fraction, with a reported IC 50 value equal to 0.11 mg/mL [20].

Insecticidal Activity
Beside all the previous mentioned studies concerning the health beneficial properties of Cachrys species, an interesting study was reported by Pascual-Villalobos and Robledo, who assessed the potential anti-insect activity of a number of Mediterranean species, including C. sicula L. The stems were extracted with three different solvents: hexane, acetone, and methanol, and samples were tested for anti-insect activity using the insect's growth and/or feeding inhibition. The insecticidal activity was tested on the stored grain pest Tribolium castaneum Herbst. Cachrys samples were incorporated into the diet at 0.05% concentration and larval growth inhibition and mortality were measured. A repellency bioassay was also performed. C. sicula hexane extract showed insecticidal activity on Tribolium castaneum Herbst. Moreover, a repellent index value over 50 (over a range from 0 to 100) was observed, indicating that Tribolium castaneum larvae were repelled compared to the control after 2 and 24 h [56].
The available data on the biological activity of Cachrys spp. are reported in Table 3.

Phytotoxicity and Toxicological Profile
C. pungens was also interestingly investigated for its potential use in sustainable agriculture, as a source of natural compounds useful for weed control. The aerial parts methanolic extract and its fractions were tested in vitro on the seed germination and root elongation of Lectuca sativa L. The most active samples, the chloroform and ethyl acetate fractions, were also tested for their effects on the common weeds Lolium perenne, Amaranthus retroflexus, and Echinochloa crus-galli. Amaranthus retroflexus was particularly sensitive to C. pungens samples, with ED 50 values equal to 2.92 and 3.21 mg/mL for the chloroform and ethyl acetate fractions, respectively [38].
Some skin damage has been related to plants belonging to Cachrys genus. Ena and colleagues, already in 1989, investigated the phytochemical content of C. libanotis L., focusing on the presence of furanocoumarins, considered the commonest non allergic phototoxic agents. The authors described some cases of phytodermatitis following contact with the flowering plant [57]. In 1991, the same authors identified from the alcoholic extract of C. libanotis the furanocoumarins 5-methoxy-, 8-methoxy-, and 5,8-dimethoxypsoralen as the substances responsible for the observed phytodermatitis [58].

Other Cachrys Species
Different studies have focused on the phytochemical and biological properties of some Cachrys species which, unlike the previous classification, are currently considered members of the Prangos genus. The Cachrys group, actually, is divided into several genera, including Cachrys, Prangos, Alocacarpum, Bilacunaria, Ferulago, Diplotaenia, Eriocycla, and Azilia [59,60].
Abad and coworkers described the biological potential of some furanocoumarins isolated from Cachrys trifida Mill [62]. Beside the name, this species is actually a synonym of Prangos trifida (Mill.) Herrnst. & Heyn, and it is currently considered a member of the Prangos group [3]. Abad and colleagues evaluated the potential role of three naturally occurring compounds in this species, namely imperatorin, isoimperatorin, and prantschimgin, in inhibiting some macrophage functions involved in the inflammatory process. The three furanocoumarins were assayed in two experimental systems: calcium ionophore A23187stimulated mouse peritoneal macrophages, a source of COX-1 and 5-LOX enzymes, and elicited mouse peritoneal LPS-stimulated macrophages, for testing the inhibitory properties on COX-2 and iNOS. All the tested furanocoumarins were effective on 5-lipoxygenase. Furthermore, imperatorin and isoimperatorin inhibited the COX-1-and COX-2-catalyzed release of prostaglandin E 2 , and imperatorin also inhibited nitric oxide production [62].
The phytochemical content of the essential oil from the same species, C. trifida (Prangos trifida) was also assessed by Palá-Paúl and colleagues [63].
One of the most investigated species previously considered a Cachrys species, and at present belongs in the Prangos genus, is Cachrys ferulacea Calest., a synonym of Prangos ferulacea Lindl. [3]. This orophilous species of the eastern Mediterranean and western Asia has been investigated for its phytochemical content, with coumarins and flavonoids being the main compounds, and different biological properties have also been verified, as highlighted by Bruno and coworkers [59].
Bertoli and colleagues characterized the essential oils from the whole fruits, seeds, and pericarps of Prangos ferulacea Lindl (synonym of Cachrys ferulacea). α-Pinene was the main identified terpene in the fruit and pericarp essential oils, being present in percentages of 18.2% and 14.2%, respectively, while sabinene was the most abundant compound in the seed essential oil (18.7%), followed by limonene and α-pinene (18.5 and 18.4%, respectively). The coumarin osthol was only assessed in the fruit and pericarp essential oils, in percentages equal to 0.8% and 1.0%, respectively [64]. Camarda and colleagues investigated Cachrys ferulacea fruits collected in Sicily, southern Italy, extracted through Soxhlet in ethyl acetate. Here, osthol, bergapten, imperatorin, and isoimperatorin were isolated and identified [65].
Badalamenti and coworkers investigated the essential oils from the leaves and flowers of a local accession from Sicily (Italy), and the phytochemical investigation they carried out allowed them to identify a new chemotype, characterized by a large amount of (Z)β-ocimene. Moreover, the essential oils were proven to exert a good antioxidant activity, inducing a decrease in ROS and an increase in the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) in opsonized zymosan (OZ)-stimulated human polymorphonuclear cells (PMNs) [68]. Other studies focused on the phytochemical content and/or the biological properties of this species. Bagherifar and coworkers, for example, analyzed the chemical composition of the essential oils from different Iranian accessions. The same authors also reported a good radical scavenging activity for these samples [69].
Moreover, C. ferulacea's potential in vitro anti-inflammatory activity was evaluated in RAW 264.7 cell lines stimulated with LPS by evaluating the presence of pro-inflammatory cytokines and mediators in the cell medium. The aerial parts extract obtained with the Naviglio ® extractor significantly inhibited the production of the pro-inflammatory mediator NO, and down-regulated the expression of p-JAK2 in a dose dependent-manner, making it the most effective species for anti-inflammatory activity, when compared to C. libanotis and C. pungens [9]. The same aerial part extract was also investigated for its furanocoumarins content with GC-MS analyses, with xanthotoxin, bergapten, and isopimpinellin accounting for 0.42 ± 0.05, 0.57 ± 0.04, and 0.050 ± 0.001 mg/50 mg of the extract, respectively [9]. Moreover, according to Dokovic and colleagues, 3,5-nonadiyne, a compound isolated from Cachrys ferulacea root essential oil, inhibited NO production in macrophages in a dosedependent manner, with an IC 50 value equal to 6.7 ± 0.8 µM, without showing any toxicity in rats [70].

Concluding Remarks and Future Perspectives
The taxonomy of the Cachrys genus is complex and the classification has been somewhat confused. For this reason, some papers reporting in their title the name "Cachrys" focused on plants species that, after more than one revision of the taxonomy, are currently included in different genera. According to the present classification, this review paper focused on the phytochemical properties and the demonstrated biological activity of the currently recognized Cachrys species, and on the whole, 34 papers were included. The analysis of the literature showed an increasing interest towards these plants, which showed, beside other phytochemical classes, a high content of terpenoids and coumarins, particularly furanocoumarins. To date, the reported phytochemical investigations allowed the identification of 177 structurally different components. The remarkable furanocoumarins content which characterizes Cachrys species is particularly interesting, as these compounds are acquiring particular importance in photodynamic therapy, because of their photosensitizing properties, and as a consequence, they appear to be a potential source of new drugs in different fields of application, such as dermatology and oncology. On the other side, some toxic effects, e.g., phytodermatitis, have been related to contact with Cachrys species due to the presence of these same compounds.
Based on the available findings, Cachrys appears to be a promising genus, with some potential health beneficial properties. Different biological properties have been reported to date for these species, such as antioxidant, antimicrobial, and anti-inflammatory effects, cytotoxic and photocytotoxic potential, as well as enzyme inhibitory properties. Together with these health beneficial properties, insecticidal effects have also been reported for one species, namely C. sicula L. However, the analysis of the literature allowed us to highlight the limited number of studies concerning the biological properties of this genus. The antioxidant and the antimicrobial effects were the most investigated properties, but just seven papers focused on the antioxidant effects of the Cachrys species and just five reports dealt with the antimicrobial potential. On the other side, despite some cases of phytodermatitis having been reported, the toxicological aspects are yet to be properly explored.
Moreover, some of the plants belonging to this genus have never been investigated. Such species remain to be explored both for their chemical and pharmacological properties. For all these reasons, this genus appears to be mostly understudied, and, potentially, a number of other interesting biological activities could still be discovered, as well as interesting secondary metabolites.
In conclusion, further in-depth research on Cachrys species is needed. Additional studies should be aimed at identifying the effective phytoconstituents of crude extracts responsible for the observed biological effects, by bioactivity guided isolation. Furthermore, toxicology and pharmacokinetic studies should also be useful in the future investigations.
Moreover, the "omics" technologies, including genomics, transcriptomics, and metabolomics, could allow even better elucidation of the biomarkers of different species, and of the relationships between metabolite biosynthesis and genetic evolution of a genus with such a difficult taxonomy. Species belonging to the same genus usually have some close chemical and biological characteristics. So, the study of the links between these properties and phylogeny could offer important information about Cachrys species and their potential therapeutic applications [71][72][73].