The Metabolome of the Ocotea spp.: From Biosynthetic Aspects to Bioactive Chemical Scaffolds by Integrating the Genus Chemical Database (OcoteaDB)

Abstract

The genus Ocotea is a significant source of bioactive agents within the Lauraceae, yet it remains underexplored. Despite its ethnomedicinal relevance and chemical diversity, Ocotea species remain taxonomically challenging as they belong to the multifaceted “Ocotea complex”, a phylogenetically unresolved group. Moving beyond previous literature Ocotea surveys, this review provides the first curated, genus-specific data set of both volatile and nonvolatile specialized metabolites. Covering research articles from 1830 to 2025, this review provides the most comprehensive synthesis to date, documenting 984 unique chemical compounds across 115 species. This effort culminated in the construction of the Ocotea Chemical Database (OcoteaDB), a novel digital resource created to streamline the genus’s chemical diversity for the global scientific community. Furthermore, it presents current knowledge of biosynthetic routes leading to key bioactive scaffolds, including aporphinoid and benzylisoquinoline alkaloids, lignoids, glycosylated flavonoids, and a diverse array of terpenoids, while addressing the stereochemical and structural intricacies unique to the genus. This review lays a robust foundation to drive future Ocotea-focused bioprospecting studies and guides research in the fields of natural products, chemophenetics, metabolomics, and medicinal chemistry regarding the Ocotea species, its chemical scaffolds, and specialized metabolites.

Introduction

The Ocotea Aubl. genus belongs to the Lauraceae Jussieu, popularly known as the laurel family, which comprises more than 3,000 species distributed among 55 genera worldwide. , The Lauraceae is among the five most abundant families in tropical and subtropical forests worldwide, classified into the Magnoliid clade as part of the order Laurales. − Several Lauraceae plants have traditional culinary usesfor example, laurel (Laurus nobilis) is used as a flavoring agent to season food, and cinnamon (Cinnamomum verum) is an important spice. Persea americana is a tree that produces one of the most important tropical fruits in the world, popularly known as avocado. − In general, Lauraceae species comprise aromatic trees, several of which are recognized as natural sources of essential oils. Examples include Aniba rosaeodora, known in Brazil as “pau-rosa”, and Ocotea catharinensis, known as “canela-preta”, both well-known for their high content of linalool, a highly sought-after monoterpenoid for enhancing woody fragrances in several commercial products. ,−

Ocotea is the largest genus of the Lauraceae, comprising more than 400 species, and several species have been documented as popular medicines, known for their significant antioxidant, analgesic, anti-inflammatory, and antitumor properties. ,, O. quixos, for example, is traditionally used as an anesthetic for wounds, skin and stomach aches. , Other species, such as O. porosa, O. odorifera, and O. catharinensis, are known as a source of woody of high-quality desirable timber, which have been used in carpentry and civil constructions for decades in South America. ,

Phytochemically, Ocotea spp. species are frequently composed of alkaloids, lignoids, flavonoids, and terpenoids. To date, only 115 species (∼28.5%) have been investigated for their chemical content in classical or modern Natural Products (NP) studies, including our recent comprehensive metabolic profiling study (Supplementary Tables S1, S1.1, and S1.2). The majority of Ocotea species are neotropical, found mainly in Central and South America, though some species are also found in Western and Southern Africa, and in Macaronesia, Madagascar, and the Comoro Islands. ,, In South America, the genus Ocotea is the third most common in the Amazon biome, which offers the largest biodiversity hotspot in the world. In the main Brazilian biomes (Amazon Rainforest, Atlantic Rainforest, Cerrado, Caatinga, and Pampas), a total of 176 species have been registered, of which 112 are considered endemic. It is estimated that more than 150 species remain to be located, taxonomically identified, and phytochemically investigated. ,

Botanically, Ocotea is considered an intriguing genus that is, at present, classified as belonging to the Persea-Laurea clade. Phylogenetically and based on DNA sequencing markers and morphological features, Ocotea belongs to the Ocotea complex sensu, which currently includes approximately 950 species of 17 distinct genera. , The Ocotea complex is of significant interest to NP chemists due to diverse biological activities, such as anti-inflammatory, antibacterial, antiviral, and insecticidal properties. However, when DNA material is unavailable for comparison, the taxonomic identification of Ocotea species based on only morphological characteristics is challenging, and these can be misclassified as species from other genera of the Ocotea complex, such as Nectandra, Persea, Licaria, and Litsea. Thus, Ocotea is considered one of the most challenging genera for species identification within the Lauraceae family. ,,,, Plant systematics, including chemical and molecular aspects, aids in the taxonomic classification of Ocotea. ,, However, despite significant ethnomedicinal, ecological, and economic relevance of this genus, the chemical space of many species is still uncharacterized. Besides, unfortunately, several Ocotea spp. are currently threatened with extinction, highlighting the importance of efforts to protect them.

Biosynthesis, Chemical Aspects, and Bioactivities

The first academic medical report on Ocotea was published in 1830, noting the therapeutically valuable oil of an Ocotea tree native to South America. Various parts of the tree were used to relieve spasmodic complaints, convulsions, and cramps, while effective against acute and chronic inflammation in the treatment of cutaneous eruptions or rheumatic pain was also reported. Data regarding the chemical aspects of Ocotea oil were first published in 1844, along with the coinage of the term “oil of laurel turpentine” in the literature. Likewise, the first nonvolatile compounds reported for Ocotea were lignans, including sesamin (pseudocubebin), isolated in 1916 from the bark of O. usambarensis, a traditional medicinal plant used by African natives in the Kimboza Forest Reserve of Tanzania. ,

In the past 70 years, and notably in the current 21st century, a large number of scientific papers have demonstrated that Ocotea is a promising bioactive genus with relevant antimicrobial, larvicidal, anesthetic, anti-inflammatory, and antitumor properties. ,,− Concerning its phytochemistry, the genus Ocotea is perceived as significantly diverse, with a high degree of chemical diversity along with its specialized metabolites. Alkaloids are the most frequently occurring chemical class in the genus, and a series of different subclasses has been observed, including a wide range of aporphines and benzylisoquinolines and the less common morphinans and phenanthrenes. ,,, The most shared alkaloid classes in the Ocotea genus consist of isoquinoline derivatives, with aporphines making up the majority (Figure ).

1.

Schematic shikimate pathway: PEP and E4P → shikimic acid → chorismic acid → aromatic amino acids → tyrosine, which yield benzyltetrahydroisoquinoline and aporphine alkaloids scaffolds.

Aporphine alkaloids serve as key chemotaxonomic markers of the genus and constitute one of the major compounds of anti-inflammatory, antimicrobial, and antiparasitic activities. ,− Succeeding the alkaloids, the lignoids and flavonoids are the most commonly described NP in the Ocotea genus, which will be covered in this review. The metabolome of Ocotea also includes volatile alkylphenols, aldehydes, terpenoids, phenylpropanoids and other less common metabolites such as coumarins, tannins, butenolides, saponins, glycosides, benzopyrans, and steroids (Supplementary Tables S2–S7). ,,−

More recently, using modern analytical molecular methods, several metabolites could be chemically discriminated in a given taxona chemophenetic approach, which can be considered a modern extension of chemotaxonomy. Metabolomics associated with DNA sequencing can also aid in solving phylogenetic issues. In addition, metabolic fingerprints can enable the identification of chemical markers within the Ocotea genusfor example, metabolites from the classes of morphinans and O-glycosylated flavonoids can be used for metabolic fingerprint discrimination within the genus. The flavone:flavonol ratio can also assist in evolutionary lineage pattern studies, once they are linked to the species specialization index, and their chemical presence can be associated with genetic modifications during the evolution of these plants. A recent application of that includes the use of the metabolic fingerprint content to avoid taxonomic errors in laboratories and industry arising from morphologically similar botanical features. In particular, metabolomics can also support a more rapid discovery of known or unknown bioactive metabolites via high-throughput screenings. −

Thus, a robust chemical database associated with the main biosynthetic pathways of the Ocotea metabolome can facilitate the recognition of new potential biomarkers and achieve more accurate and faster species differentiation. Herein, we present and discuss the most comprehensive chemical review of the Ocotea genus. The main objective is to provide an updated version of current knowledge on the chemical and biosynthetic characteristics of the Ocotea metabolome to offer additional insights and orient further studies targeting Ocotea species, culminating in the creation of the Ocotea chemical database (OcoteaDB). It is also intended to provide easy access to chemical cores and unique metabolites, integrated with main biosynthetic routes to access their chemophenetic value.

Ocotea spp. Specialized Metabolites

Alkaloids, lignoids, and terpenoids are the main representative classes found, and flavonoids are also found, though in a smaller number of Ocotea spp. ,,, Despite notable discoveries by Professor Otto R. Gottlieb (University of São Paulo, deceased in 2011) and collaborators between 1960 and 1980, research interest has only recently shifted back to the genus, with an increased number of relevant publications in the past 15 years. ,, In this review, a total of 984 metabolites were compiled into the OcoteaDB, based on data retrieved from both the scientific literature (1830 and 2025) and NP databases, including: Web of Science, ScienceDirect, Google Scholar, Dictionary of Natural Products (DNP^©), KNApSAcK, and the Nuclei of Bioassays, Biosynthesis and Ecophysiology of Natural Products Database (NuBBE). Inclusion criteria were limited to peer-reviewed studies identifying volatile and nonvolatile Ocotea compounds via Nuclear Magnetic Resonance (NMR) and liquid chromatography coupled to mass spectrometry (LC-MS), or gas chromatography-MS (GC-MS). Duplicates were manually checked and filtered, while inconsistencies in chemical nomenclature were resolved using the IUPAC standard and cross-references with PubChem, ChemSpider and SciFinder. The OcoteaDB includes detailed information such as chemical and/or trivial names, molecular formulas, monoisotopic masses, metabolite scaffold, biosynthetic classes, SMILES, Ocotea species where it was identified, and respective bibliographic references (see Supplementary Tables S2–S7).

Alkaloids

The main alkaloid class produced by Ocotea comprises the aporphines. This class is not exclusive to Lauraceae, as it also occurs in other plant families, such as Magnoliaceae, Menispermaceae, Papaveraceae, Ranunculaceae, Hernandiaceae, and Annonaceae. , Aporphine alkaloids are one of the largest groups of isoquinoline alkaloid derivatives, with more than 500 known representatives in the literature. Despite the chemical diversity and distribution of this class, robust data regarding its metabolic biosynthetic pathways in the literature are sparse. ,

The biosynthesis of the aporphine core includes a Pictet–Spengler reaction type, where a Mannich-like condensation takes place to join dopamine and 4-hydroxyphenylacetaldehyde precursors by amino alkylation and form (S)-norcoclaurine (1), the primary precursor for the benzylisoquinoline class of compounds. O-methylation of 1, yielding (S)-coclaurine (2), is mediated by norcoclaurine 6-O-methyltransferase (E1) and S-adenosyl methionine (SAM), common substrates involved in the stereoselective mechanism of methyl group transfers via a S_N2-type reaction. Subsequently, precursor 2 is catalytically converted into (S)-N-methylcoclaurine (3) by coclaurine N-methyltransferase (E2). Following the pathway, alkaloid 3 can be hydroxylated at position 3′ of the benzylic ring to form the tetrahydroxy-substituted pattern in the presence of the enzyme N-methylcoclaurine 3′-hydroxylase (E3), to afford (S)-3′-hydroxy-N-methylcoclaurine (4). In addition, the hydroxyl group at position 4′ can be methylated by 3′-hydroxy-N-methylcoclaurine-4′-O-methyltransferase (E4)/SAM to afford the alkaloid (S)-reticuline (5), which is one of the main benzylisoquinoline alkaloids found in the Ocotea genus. For instance, alkaloid 5 has been isolated from several species, including O. lancifolia, O. caparrapi, O. odorifera, and O. caudata, among others. ,, The biosynthetic pathway is depicted in Figure .

2.

Biosynthetic pathway of benzylisoquinoline alkaloids 1–5. E1-norcoclaurine 6-O-methyltransferase; E2-coclaurine N-methyltransferase; E3-N-methylcoclaurine 3′-hydroxylase; E4-3′-hydroxy-N-methylcoclaurine-4′-O-methyltransferase.

Although Ocotea does produce R isomers as well, the dominant presence of S isomers in the Ocotea genus has been described for benzyltetraisoquinoline alkaloids. The biosynthesis of either the R or S stereoisomers after the Mannich reaction is expected to be dependent on the configuration of the precursor. For instance, previous studies reported (6R)-reticuline (8) from O. diospyrifolia, while (6S)-reticuline has been obtained from O. odorifera. , In addition, different (R)-alkaloid isomers have been reported from a few other Ocotea species, such as O. caesia, O. lancifolia, and O. velloziana. ,, These findings corroborate the significance of chemophenetics and biosynthetic studies, which, to date, are poorly described in the literature. This area can, therefore, be further explored by using specific enzymes and substances with atom labeling to confirm the assumptions and observations highlighted in this review. Furthermore, alkaloid 5 can undergo phenolic oxidative coupling, which plays a significant role in modifying the basic benzyltetrahydroisoquinoline skeleton to yield several other types of alkaloids, such as aporphines and morphinan derivatives. While the former is widespread in the Ocotea genus, the latter is also present, though in fewer Ocotea species. ,, The aporphines (S)-corytuberine (6) and (S)-isoboldine (7) have been identified in different Ocotea species, such as O. caesia, O. caudata, O. lancifolia, and other Lauraceae genera. ,, Literature data , support the change in the absolute configuration of 5 to the R enantiomeric form (8), which can be achieved by an oxidation–reduction process via the formation of the 1,2-dehydroreticulinium cation intermediate (9). The R enantiomer is produced via oxidation, which turns it up in a planar iminium cation, enabling the formation of 8. These steps allow for the stereochemical change through the action of two coordinated enzymes: 1,2-dehydroreticuline synthase (E5)/NADP⁺, which is responsible for the first heterocyclic nitrogen oxidation, and 1,2-dehydroreticuline reductase (E6)/NADPH, reducing the ion to form the enantiomeric alkaloid 8. The classes of morphinans found in Ocotea are predominantly biosynthetically derived from the (R)-reticuline (8) stereoisomer. ,, The Ocotea spp. are not known to be morphinan producers as species from Papaveraceae; however, methylated morphinans such as pallidine (9) have already been isolated from O. acutangula, O. acutifolia, and O. lancifolia. Curiously, the most known morphinans, such as codeine, salutaridine, and morphine, have not yet been found in Ocotea spp. Biosynthetically, morphinan alkaloids are produced by reactions mediated by coupling enzyme synthases that are cytochrome P450-dependent monooxygenases, such as salutaridine synthase (E7), responsible for giving rise to the dienone morphinan alkaloids (Figure ).

3.

Biosynthetic pathway of aporphine, morphinan alkaloids 6–9 and related compounds from (S)-reticuline (5). (A) Oxidative coupling of (S)-reticuline to forming aporphine alkaloids; (B) isomerization of (S)-reticuline (5) to (R)-reticuline (8); (C) Conversion of (R)-reticuline (8) into salutaridine and morphinan derivatives. E5-1,2-dehydroreticuline synthase; E6-1,2-dehydroreticuline reductase; E7 - salutaridine synthase.

Moreover, the CYP complex enzyme CYP80A1 catalyze the C–O intermolecular phenol-coupling reaction between benzylisoquinoline cores to afford a dimer class of alkaloids, the bisbenzylisoquinolines. Bisbenzylisoquinolines occur in plant families such as Lauraceae, Menispermaceae, Berberidaceae, and Ranunculaceae, but they are not characteristic of Ocotea spp., where they are rarely reported. Within Ocotea, the bisbenzylisoquinolines, tetrandrine and thalmine have only been described in two species to date, O. rodiei and O. venenosa, both of which were historically assigned to the genus, but later reclassified as Chlorocardium rodiei and C. venenosum based on morphological and phylogenetic evidence. ,,

Alkaloid Profile

In their comprehensive review using multivariate statistical data analysis, Antonio et al. (2020) claimed that approximately less than 10% of Ocotea species have been chemically recognized for their alkaloid content. Their study employed 31 alkaloid-producing Ocotea species, as extensively described earlier by Teles et al. (2019). The present review has updated this number to 60 Ocotea species reported to date for eliciting biosynthetic pathways for alkaloid production. Thus, the alkaloid class stands as the most prevalent class among the investigated Ocotea species, with 156 curated reported and well described metabolites (Supplementary Tables S2 and S2.1). Regarding only classical phytochemistry studies, aporphine alkaloids constitute the most common alkaloid source isolated from Ocotea spp., with 107 unique representative structures in the literature (Figure ).

4.

Chemical diversity of alkaloids in the Ocotea genus is expressed as the number of different chemical structures reported in the literature for each subclass.

From the 60 alkaloid-producing species, 55 (∼91.5%) had the aporphine core described in at least one study (core C2). Moreover, beyond the aporphines, classes such as their precursors or derivatives that include benzylisoquinoline, proaporphine, phenanthrene, morphinans, and protoberberines have been reported in a fewer number of Ocotea species, with a total of 43 unique metabolites (cores C1 and C3–C6). Even less common chemical cores have been reported; for example, aminoaporphine (core C7) has been isolated from O. variabilis, the indole core (C8) from O. minarum, the pyrrolidine core (C9) from O. caudata, and the isoquinoline core (C10) from O. diospyrifolia. ,,, Additionally, the isoquinoline core has been recently annotated in the O. delicata. The different alkaloid chemical scaffolds found in Ocotea species are displayed in Figure .

5.

Alkaloid chemical scaffolds found in Ocotea species: (C1) benzylisoquinoline, (C2) aporphine, (C3) proaporphine, (C4) phenanthrene, (C5) morphinan, (C6) protoberberine, (C7*) aminoaporphine, (C8*) indole, (C9*) pyrrolidine, and (C10*) isoquinoline alkaloids from the Ocotea genus. *Only one study in the literature has reported the isolation of the chemical core. Note: depending on each class scaffold, R = H/H₂/O^–/CH₃.

Aporphines are a relevant alkaloid class for evolutionary lineages in Lauraceae and are considered a biomarker for discriminating between Ocotea as a basal genus and other genera in the family. For example, the patterns of the substituted aporphines can aid in the differentiation of Ocotea from Cinnamomum and even other genera placed at the unresolved Ocotea complex sensu. , Thus, an alkaloid fingerprint profile including aporphine patterns can support phylogenetic studies of Lauraceae, providing significant key information to elucidate alkaloid expression, complementary to gene sequencing. There are different subclasses of aporphine in Ocotea species, such as dihydroaporphines, didehydroaporphines, oxoaporphines and didehydroxoaporphines. However, within the Ocotea genus, only specific substituted aporphines can serve as significant differential biomarkers, especially those with high levels of oxidation (e.g., 3-hydroxydicentrine or dehydroocoteine). This is expected, as the majority of Ocotea species are aporphine producers and share the same biosynthetic pathway. Thus, besides the potential for genus differentiation within the Ocotea complex, other metabolite classes should be utilized as a more efficient biomarker selection for a chemosystematics of the Ocotea genus.

Phylogenetic and evolutionary classification states that Ocotea species producing majoritarian aporphines are part of the Old World, and thus, it is assumed that they are more basal in the Lauraceae lineage. , On the other hand, Ocotea species that produce more evolved alkaloid cores, such as the morphinan, could be considered to be from the New World, as they have evolved from specialized biosynthetic pathways; for example, in O. acutangula, where six different morphinan alkaloids have been isolated. Only five Ocotea species (O. acutangula, O. acutifolia, O. brachybotrya, O. caudata, and O. lancifolia) have been described in the literature as morphinan producers. ,,,,,

Furthermore, chemometrics and metabolomics studies can aid in the search for specific biomarkers and alkaloidal fingerprints to facilitate species differentiation. For example, plants from the Licaria and Nectandra genera might be taxonomically confused with those from the Ocotea genus due to their botanical similarity in terms of leaf and flower morphology. Some of these species can share the same stomatal surface shape, making morphological differentiation difficult. Thus, plants such as those from the Ocotea genus can be more easily differentiated by their chemical profiles and through biomarker comparisons when accompanied by chemometrics analyses of their specialized metabolites. Chemical profile data, together with respective morphological features and phylogenetic studies, can avoid inaccurate taxonomic assumptions and incorrect genus identifications. ,

Regarding its chemistry and bioactivity, aporphine cores exhibit diverse chemical possibilities in terms of substituents, including methoxy, methylenedioxy, and hydroxy substituents attached at different positions to the aromatic ring. ,, This review describes all substituted aporphine cores within the Ocotea genus reported in the literature, including the derivative classes of oxoaporphines, proaporphines, O-aporphines, dihydroaporphines, didehydroaporphines, and phenanthrenes (Figures and ). The detailed identifications of the chemical compounds, including their monoisotopic masses, molecular formulas, chemical names, SMILES, and Ocotea species from which the metabolites were isolated (Supporting Information file, Tables S2–S7).

Furthermore, aporphine alkaloids (C2) from different Ocotea species have been reported with pronounced anti-inflammatory and antineoplastic properties. Examples include the aporphine boldine, the most prevalent scaffold found in O. lancifolia, O. spixiana, and other Ocotea spp. and Lauraceae species, which can induce antipyretic effects possibly due to the effect on the COX pathway, leading to the inhibition of prostaglandin’s release. , Likewise, the aporphine alkaloid glaucine, which occurs in O. vellosiana and O. macrophylla, exhibits interesting anticancer properties by suppressing the activity of nuclear factor kappa-activated B cells (NF-κB). In addition, the glaucine congener has also demonstrated a reduction in metastatic breast cancer cell invasion. Recently, nuciferine-derived aporphine alkaloids were associated with O. villosa cytotoxicity against MCF-7 human breast cancer cells.

The aporphine alkaloid dicentrine and the oxoaporphine dicentrinone have demonstrated a broad spectrum of biological activities, such as the inhibition of topoisomerase I and II enzymes, which are relevant therapeutic targets of current chemotherapy protocols. In addition, more recent data have confirmed the significant in vivo antinociceptive effects of dicentrine. The mechanism of action includes attenuating mechanical and cold hypersensitivity under inflammatory conditions via the activation of the transient receptor potential TRPA1. Dicentrinone has also shown potent antiparasitic activity against the trypomastigote forms of Trypanosoma cruzi and significant changes in lipid biological surfaces with reduced mammalian cytotoxicity. , Moreover, glaziovine, a rare proaporphine isolated from O. glaziovii, exhibit remarked anxiolytic effects and was indeed registered as a tranquilizer under the trademark Suavedol by the pharmaceutical Simes S.p.A., Milano. −

Regarding the benzylisoquinoline alkaloids (C1) (Figure ), R-coclaurine has demonstrated a highly pronounced anti-HIV activity, with an EC₅₀ value of 0.8 μg/mL. , Furthermore, promising butyrylcholinesterase inhibition activities have been reported for two other benzylisoquinoline alkaloids: (+)-reticuline, with an IC₅₀ value of 33.6 ± 3.0 μM, and (+)-N-methylcoclaurine, with an IC₅₀ value of 15.0 ± 1.4 μM. The O. odorifera leaf decoction is rich in alkaloid (+)-reticuline, which, after alkaloid isolation and evaluation, exhibited a significant antiedematogenic effect together with neutrophil recruitment inhibition in a dose-dependent manner, suggesting that both the COX and LOX inflammatory pathways were inhibited. , Benzylisoquinolines and aporphines are the major classes isolated from Ocotea species and can be considered major bioactive NP in the genus. They are encountered in several different Ocotea species, such as O. puberula, O. vellosiana, O. acutifolia, O. macropoda, O. leucoxylon, O. discolor, O. caesia, O. odorifera, O. diospyrifolia, O. lancifolia, and O. brachybotrya, among others (Supplementary Tables S2 and S2.1). Besides, other members of the broader benzylisoquinoline/isoquinoline alkaloid space have reached the pharmaceutical market, even if they were not isolated from Ocotea spp. In particular, apomorphine, an aporphine alkaloid, is an established drug for Parkinson’s disease, and papaverine, a benzylisoquinoline alkaloid, has long been used clinically as a vasodilator/antispasmodic.

Ocotea spp. Lignoids

Since the identification of the highly bioactive lignan podophyllotoxin in 1933, the biosynthetic pathways of lignoids have been thoroughly elucidated, enabling a clear understanding of their core bioactive scaffolds, such as the podophyllotoxin 8–8′, 2–7′ aryltetralin lactone derivatives. Although some gaps persist in the literature regarding the most specific biosynthetic steps, particularly for the more complex neolignans, which exhibit vast structural and stereochemical diversity. ,, Given their pharmacological relevance, podophyllotoxin and related aryltetralin lignans have become central targets of synthetic, biosynthetic, and medicinal-chemistry investigations. Notably, the biosynthetic machinery responsible for assembling this characteristic lactone scaffold has been identified in several plant families, including Lauraceae, Cupressaceae, and Linaceae. Among Lauraceae, the aryltetralin lignan (−)-morelensin, isolated from O. macrophylla, shares the same 8–8′, 2–7′ biosynthetic origin as podophyllotoxin but differs structurally by possessing only two methoxy substituents on its aromatic rings, in contrast to the three groups in podophyllotoxin. ,−

Regarding the chemical cores of lignoids, their general structural classification is based on the nature of the intermolecular linkage that joins the two phenylpropanoid (C₆–C3) units. These linkages may occur through carbon–carbon (C–C) bondsas observed in lignans and neolignansor through carbon–oxygen–carbon (C–O–C) bridges, giving rise to oxyneolignans. ,, Among the Ocotea species reported in phytochemical studies in the literature, ∼11.5% (n = 21) produce lignans, while ∼87% (n = 158) have at least one neolignan in their metabolome. Oxyneolignans are less frequently encountered, occurring in only ∼1.6% (n = 3) of studied species, including O. cymosa and O. costulatum. This profile suggests an evolutionary shift or specialization within Ocotea toward neolignan-dominated metabolism, possibly a result of genomic expansion of biosynthetic genes regulating phenylpropanoid coupling. Ocotea species express a broad set of oxidative enzymes (e.g., peroxidases, laccases) and dirigent proteins (DIRs) that guide the stereoselective radical coupling of monolignols into nonclassical C₆–C₃ dimers of the neolignan scaffolds. Unlike strict 8–8′ linkages in lignans, these enzymes favor alternative coupling positions (8–1′, 8–3′, 7–3′, etc.), enabling formation of complex neolignan scaffolds such as benzofurans and bicyclic octane cores.

Regarding the lignan scaffolds, the 7–7′ epoxylignans (furan lignan, C2, Figure ) and 9–9′ diepoxylignans (furofuran lignan, C3) represent the most abundant scaffolds in the Ocotea genus, together accounting for approximately 76% of all lignans reported for Ocotea species in the phytochemical literature. The derivatives of the bicyclic octane neolignans are encountered in over 60% of the lignoids Ocotea species, such as O. macrophylla, O. catharinensis, O. porosa, O. cymosa, and others (Supplementary Table S3). − Lignoids in general represent the second most common class of specialized metabolites in the Ocotea genus, characterized in 18 out of 115 different Ocotea species already chemically studied. ,− ,− Our recent publication, which employed modern chemical annotation for screening several Ocotea species not chemically profiled before, increased the number of known Ocotea species able to elicit lignoid biosynthesis to 45 (Supplementary Tables S3 and S3.1).

6.

C₆–C₃ units of a phenylpropanoid core and examples of the difference between lignans and neolignans based on different potential linkages.

Figure illustrates Haworth’s (1937) definition of a lignan, which remains widely accepted today. According to this definition, lignans arise from the oxidative dimerization of two C₆–C₃ units connected at the 8–8′ (β–β′ linked) positions, with possible variations at the 7–7′, 9–9′, and 2–7′ positions. Linkages deviating from this pattern, such as 3–3′, 7–1′, 7–3′, 8–1′, and 8–3′ linkages or ether linkages such as 8-O-4′, are classified as neolignans. ,,

Later, Gottlieb (1974) suggested that the classification can be based on their respective biosynthetic precursors, helping to rationalize the formation of different coupling patterns. Thus, the basic lignoid core is formed through the oxidative coupling of phenylpropanoids, yet the plant’s metabolism shows a degree of biosynthetic independence, resulting in various congeners. The term “lignans” should be retained for derivatives condensed via the oxidative coupling of cinnamyl alcohol and/or cinnamic acid (8–8′ linked). Neolignans, on the other hand, are derived via condensation and oxidative coupling between propenylphenols and allylphenols, typically via non-β–β′ linkages. More recently, the Lignan Handbook published by Lewis and collaborators (2022) has argued that this distinction should be revised, recommending abandonment of the prefix “neo-” because these compounds arise from the same phenylpropanoid biosynthetic origin and differ mainly in their coupling patterns. This view echoes earlier considerations by Gottlieb that they are not “different pathways”, but different substrate pools + oxidative coupling. Previously, Gottlieb and Yoshida (1984) had indicated a similar perspective, but at that time, the academic community, particularly organic synthetic chemists, was already employing the neolignoid nomenclature.

In the present review, the traditional lignan and neolignan terminology was retained to preserve consistency with the primary literature and with the organization of the chemical records compiled in OcoteaDB, thereby enabling direct comparison and traceability across published reports on Ocotea spp. Nevertheless, recent proposals for revising this nomenclature are acknowledged, and the biosynthetic relationships among these lignoids are discussed here in an integrated manner. Accordingly, these classes of compounds are synthesized in plants via the shikimate pathway, utilizing as precursor the p-coumaric acid (10), which serves as a critical core in the phenylpropanoid pathway and has been isolated from O. minarum. , From this pivotal intermediate, a tightly regulated sequence of hydroxylation, methylation, reduction, and oxidative coupling reactions gives rise to the diverse lignoid scaffolds discussed herein (Figure ). Collectively, this metabolic convergence underscores the phenylpropanoid pathway as a central biosynthetic platform from which a wide spectrum of structurally and biologically distinct lignoids and related specialized metabolites emerge.

7.

Primary monomers are derived from the shikimate pathway in the biosynthesis of lignans and neolignans. Following the formation of p-coumaric acid (10), a cascade of biosynthetic reactions is initiated within the phenylpropanoid pathway.

The biosynthesis of lignoids frequently results in enantiomerically pure metabolic products, attributed to the control exerted by stereoselective enzymes during the coupling reactions that form lignans and neolignans. Examples include the oxyneolignan (S)-virolongin B (C7.2) and (S)-ococymosin (C6.3), both neolignans isolated from O. cymosa, showcasing the predominant optically active forms, similar to certain alkaloids. However, it is important to mention that these NP can also be isolated as racemic or enantiomeric mixtures. − Reactions typically catalyzed by laccase (Lac) or peroxidase (Pex) enzymes, often guided by dirigent DIR-proteins, that control the regioselectivity and enantioselectivity of the β–β′ linkage, lead to the formation of the furofuran lignan (+)-pinoresinol, a 9–9′-diepoxylignan. Subsequently, the furofuran syringaresinol (11) and yangambin (12) are derived from this through enzymatic hydroxylation and SAM/OMT methylation. Compounds 11 and 12 have been described in O. duckei, and 12 also in O. heterochroma, respectively (Supplementary Table S3; Figure ).

8.

Coniferyl alcohol radicals and DIR enantioselective formation of (+)-pinoresinol and related lignoids syringaresinol (11) and yangambin (12).

The furan lignans (e.g., 7–7′-epoxylignan cores) are formed by the coupling of propenylphenol monomer units, which are coniferyl alcohol derivatives, such as eugenol (13) and isoeugenol (14). DIR-guided radical dimerization of E-isoeugenol gives the characteristic 7,7′-epoxylignan (+)-verrucosin, which functions as a central biosynthetic intermediate in Lauraceae. Subsequent SAM-dependent O-methyltransferase reactions on this verrucosin-type scaffold generate the closely related furan lignans such as (+)-veraguensin (15) and feasibly the (+)-methylverrucosin (16), as they all share a common 2,5-diaryl-3,4-dimethyltetrahydrofuran core derived from oxidative coupling of eugenol- or isoeugenol-type propenylphenols (Figure ).

9.

Proposed (+)-verrucosin, (+)-veraguensin (15) and (+)-methylverrucosin (16) biosynthesis via isoeugenol (14) and derivative precursors.

These examples highlight the crucial role of configuration in lignoid biosynthesis, as several lignan subclasses incorporate four or more stereogenic centers formed during oxidative coupling and downstream tailoring steps. Among these structures, the furan lignan cores corresponding to compounds 11 and 12 are among the most frequently reported in the Ocotea genus isolated from O. foetens, O. veraguensis, O. catharinensis, and several additional Ocotea species (Supplementary Table S3). The conversion of coniferyl acetate in the chavicol and anol units of phenylpropene, such as eugenol and isoeugenol, is catalyzed by NADPH-dependent reductases and eugenol and isoeugenol synthases, respectively (Figure ). ,

The neolignan scaffolds are abundant in the Lauraceae, particularly in the Ocotea genus (Supplementary Table S3). However, even though lignoids display diversity, within the Ocotea genus, more than one-third (48.4%, n = 88) are bicyclic neolignoids, making this subclass the most representative lignoid subclass in the Ocotea genus. These compounds have been identified in different species, such as O. aciphylla, O. bullata, O. catharinensis, O. veraguensis, and O. porosa, among others. ,,,,, Following closely, the benzofuran subclass accounts for 31.3% (n = 57) of the isolated lignoids, found in species such as O. catharinensis, O. veraguensis, O. porosa, and O. macrophylla, among others (Supplementary Table S3). The high prevalence of these two subclasses within Ocotea spp. is not merely coincidental. Considering biosynthetic aspects, benzofuran neolignans, particularly those with 8–1′ and 8–3′ linkages, are considered key precursors in the biosynthetic pathway before rearrangement into bicyclic octane neolignoids. Figure illustrates the acid-catalyzed rearrangement of the 8–1′ linkage to form the classical bicyclic neolignoid scaffold. Additionally, the reverse pathway to the benzofuran core is feasible, allowing for further biochemical modifications within the plant biosynthesis. ,,

10.

Proposed bicyclic octane neolignoids via 8–1′ linkage radical monomers based on an acid-catalyzed rearrangement mechanism.

Lignoid Profile

Over the past century, lignoids have been extensively studied in terms of their role as phytochemical biomarkers through various systematic approaches, prominently by Gottlieb and collaborators (from 1970 to 2000). ,,,− ,,,, Recently, a metabolomics study using only literature data by Antonio et al. (2020) showed a shift in academic interest toward alkaloids in the 21st century, with lignoids receiving less attention. Nevertheless, despite their research limitations regarding the acquisition of data for metabolomics models and the limited available data on lignoids from Ocotea spp., their multivariate analysis corroborated that Ocotea lignoids are valuable for chemophenetics. In addition, according to their work, Gottlieb’s statement that Ocotea species exclusively biosynthesize either alkaloids or lignoids is no longer valid, as modern chemical analytical techniques have indicated that different Ocotea species can elicit both alkaloids and lignoids (e.g., in the case of O. macrophylla, O. duckei, and O. minarum).

In this context, the study of metabolomics features through untargeted metabolomics strategies has the potential to enhance plant taxonomic issues in the genus. Species differentiation could be rapidly achieved using metabolic fingerprints obtained from state-of-the-art analytical tools such as UPLC-HR-ESI-MS. , Antonio et al. (2020) also demonstrated that comparing lignoid contents is effective in distinguishing between similar Ocotea species due to the significant variation in the lignoid profiles. Furofuran lignans, such as 12, have proven to be effective phylogenetic biomarkers within the genus. The continued application of untargeted metabolomics may uncover new markers, aiding taxonomic classifications, as these methods are already widely used in drug and phytomedicine quality control. −

Increasing focus on the chemical data and bioactivity of lignoids over the past decade suggests that these compounds may soon regain broader interest within the academic community. Original articles accessing chemical data and the bioactivity of lignoids have increased over the past 10 years. Notably, pharmacological effects have been reported in approximately 20% of the chemically characterized Ocotea species. To date, 182 lignoid chemical structures have been isolated (and curated) from the genus (Supplementary Table S3). Figures ,,,,, present the lignoid cores found among the different Ocotea lignoid producers reported in the literature.

11.

Chemical diversity of lignoids in the Ocotea genus, expressed as the number of different chemical structures reported in the literature.

12.

Lignan chemical scaffolds found in Ocotea species: (C1) 8–8′-lignans, (C2) 7–7′-epoxylignans, (C3) 9–9′-diepoxylignans, (C4) 8.8′,2.7′-aryltetralin lignan lactones, and (C5) 2–7′-cyclolignans. The molecular diversity of lignans in the genus Ocotea is expressed by the number of different chemical structures for each subclass of lignan scaffolds.

13.

Benzofuran neolignan chemical scaffolds found in Ocotea species: (C6.1) 8.1′-, (C6.2) 8.3′- and (C6.3) 8.5′-benzofuran neolignans.

14.

Oxyneolignan chemical cores found in Ocotea species: (C7.1) dioxyneolignan and (C7.2) oxyneolignan.

15.

Lignoid chemical scaffolds found in Ocotea species: (C8.1) 8.1′-neolignan, (C8.2 and C8.2a) 8.3′-neolignan and (C8.3) 4.3′-didymochlaenone neolignans.

16.

Bicyclic neolignan chemical cores found in Ocotea species: (C9.1) and (C9.1a) 7.3′, 8.1′-connected and (C9.1b) 7.3′, 8.1′-dimer. (C9.2) 7.1′, 8.3′-connected. (C9.3) 7.3′, 8.5′-connected bicyclo[3.2.1]­octanoid and (C9.3a) 7.3′, 8.5′ oxabicyclo[3.2.2]­nonane-type neolignans.

Ocotea species-produced lignoids have exhibited a high potential for treating or preventing neglected tropical diseases (NTDs) such as dengue fever and leishmaniasis, which affect millions of people in vulnerable situations worldwide. Accordingly, potent larvicidal and antiprotozoal effects have been reported for different lignoids isolated from plants of the Ocotea genus. For example, neolignans isolated from O. cymosa have shown a high larvicidal potential, halting the Aedes aegypti life cycle at the larval stage with 100% mortality. , The lignan yangambin (12, C3) and the neolignan licarin A (C6.2) from O. macrophylla have exhibited low IC₅₀ values for amastigote and promastigote forms of different Leishmania (L.) subspecies. In addition, 12 and burchelin (C9.2) have been reported in the literature as highly cytotoxic agents against trypomastigote forms of T. cruzi. −

Moreover, lignoids are recognized as compounds with notable pharmacological properties, including pronounced antitumor, antiviral, and anti-inflammatory effects. ,,,, For instance, reports have indicated the activity of furofuran lignans (C3), aryltetralin lactones (C4), and cyclolignans (C5) against a broad spectrum of cancer cell lines, including human lung adenocarcinoma (A-549), human colon adenocarcinoma (HT-29), human breast adenocarcinoma (MCF-7), and murine lymphocytic leukemia (P-388). Regarding antiviral activities, different classes of lignoids have been proven effective against hepatitis B virus (HBV), human cytomegalovirus (HCMV), and human immunodeficiency virus (HIV), severe acute respiratory syndrome-related coronavirus (SARS-CoV), and Zika virus (ZIKV). , Additionally, diastereomeric lignans isolated from O. macrophylla have exhibited PAF antagonism and the potent dual inhibition of COX-2/5-LOX pathways. A study by our research group has confirmed the potent in vivo anti-inflammatory activity of the bicyclic octane neolignans (C9) via the dual inhibition of prostaglandin E2 (PGE2) production and neutrophil recruitment. However, they were isolated from Aniba firmula, which also belongs to the Lauraceae family. ,, The derivatives of the bicyclic octane neolignans are encountered in over half of the lignoid-producer Ocotea species, such as O. macrophylla, O. catharinensis, O. porosa, O. cymosa, and others (Supplementary Table S3). − , Furthermore, the bicyclic neolignoid sibyllenone (C9.2), isolated from O. bullata, has shown promising anti-inflammatory activity via the inhibition of 5-LOX enzymes.

Finally, Teppono et al. (2016) have reported advances in lignoid pattern biosynthesis, detailing a classification into 10 lignan core subtypes and 15 neolignan subtypes. Our review highlights that the Ocotea genus encompasses 5 of these 10 lignan subtypes: 8–8′-lignans (C1), 7–7′-epoxylignans (C2), 7–8-secolignans (C2.1), 9–9′-diepoxylignans (C3), 8.8′, 2.7′-aryltetralin lignan lactones (C4), and 2–7′-cyclolignans (C5). Additionally, the genus hosts four different neolignan core subtypes, including 8.1′-, 8.3′- and 8.5′-benzofuran neolignans (C6), 7.3′-, 8.4′-/8.4′-oxyneolignans (C7), 8–1′-/8–3′-/4–3′-neolignans (C8), and bicyclic octane neolignans (C9). Thus, alongside its alkaloids, Ocotea is also a remarkably diverse source of lignans and neolignans, offering significant variations in scaffold structures (Figures –).

Ocotea spp. Flavonoids

Flavonoids are specialized metabolites characterized by two benzene rings linked by a pyran moiety, which can undergo a wide variety of chemical modifications. These include substitutions with hydroxyl, methoxy, or glycosyl groups, which significantly affect their physical and chemical properties and their biological activities. Flavonoids also play a crucial role in plant ecology, acting as defense agents against several environmental and biotic stresses, such as ultraviolet radiation, pests, and diseases, as well as beneficial effects on human health in terms of their antioxidant, anti-inflammatory, anticancer, and antiviral properties. − In addition, these compounds represent a diverse group of naturally occurring polyphenolic metabolites prevalent in the plant kingdom, including Lauraceae and, consequently, the Ocotea genus. ,,,

In this review, flavonoids were identified as the third most commonly reported metabolite class in the Ocotea genus, with 61 different described metabolites distributed in 20 species. However, in our recent publication using modern annotation analytical techniques, this number was significantly increased to 60 Ocotea species (Supplementary Tables S1.1–1.2, S4 and S4.1).

The analysis of flavonoid chemical cores can aid in taxonomic classifications and fill potential phylogenetic gaps. By identifying specific flavonoid subclasses, investigations in the fields of chemophenetics, metabolomics, and transcriptomics can be implemented to elucidate the relationships between different Ocotea spp. and other genera. Biosynthetically, the carbon chain is named as the C₆–C₃–C₆ unit coming from both phenylpropanoid (C₆–C₃) and acetate (3 × C₂) metabolism (Figure ). , Such molecules can exhibit chirality and interesting bioactivity, and the flavanone core serves as the central precursor for downstream subclasses such as flavones and flavonols. The latter being the most abundant in Ocotea, particularly those based on kaempferol (17) and quercetin (18), with myricetin occurring less frequently, and the former apigenin (19) and luteolin (20). ,,,,

17.

General biosynthesis of Ocotea flavonoids, including flavanone (naringenin), flavonol (17 and 18), flavone (19 and 20), and flavanonol (21).

Beyond the initial transformations summarized in Figure , extensive enzymatic diversification can expand the structural complexity of flavonoids. , For example, flavonolignans represent a hybrid class of NP biosynthesized through the oxidative coupling of flavonoids such as taxifolin (21, 5,7,3′,4′-flavanonol) with a phenylpropanoid, typically coniferyl alcohol precursor. Recently, a flavonolignan, mururin A was isolated for the first time within the Ocotea genus, specifically from O. diospyrifolia.

Flavonoid Profile

In the Ocotea genus, flavonoids are predominantly derivatives of the flavonols 17 and 18. Despite growing research interest, the knowledge on Ocotea flavonoid profiles remains limited. , To date 61 distinct flavonoids have been reported, of which ∼ 18% are nonsubstituted flavonoid cores (n = 11), while the majority ∼67% (n = 41), are glycosylated derivatives; about ∼8% (n = 5) are of lactonized cores, and another ∼8% (n = 5) are catechin or flavonol dimers. Among the glycosylated ones, ∼59% of the total (n = 36) are O-glycosylated, and only ∼8% (n = 5) are C-glycosylated derivatives. O-alkylated flavonoids have so far been reported in only four species, including O. canaliculata, O. caudata, O. guianensis and O. corymbosa, whereas C-glycosylated derivatives have been found in O. porosa, O. aciphylla, O. foetens, O. nutans, and O. odorifera (Tables S4 and S4.1).

A recent publication regarding flavonoid profile using UPLC-HRMS indicated the presence of an apigenin flavone chemical core in six other Ocotea species, including O. diospyrifolia, O. guianensis, O. lancifolia, O. notata, O. odorifera, and O. porosa. The apigenin derivatives were found mainly in C-glycoside form, with O. porosa exhibiting the highest flavone:flavonol ratio. Taken together, the predominance of O-glycosylated flavonoids in the genus, the low occurrence of O-alkylated flavonoids and the annotation of C-glycosylated apigenin derivatives in different species suggest a basal or intermediate phylogenetic position for Ocotea within the Lauraceae, as previously pointed out by Antonio et al. (2020). This metabolic signature can reflect a transitional evolutionary stage, where simpler glycosylation patterns precede the complex methylation seen in other Lauraceae genera. This is supported by the high occurrence of O-alkylated flavonoids in evolutionarily advanced plant groups. However, as phylogenetic studies increase in accuracy and precision, with a larger set of evaluated species, additional studies using DNA barcoding together with morphological data could provide further evidence to support or refute this hypothesis.

Flavonoids are not only crucial from an evolutionary and phylogenetic perspective but are also noted for their diverse bioactivities. The most common flavonoid core found in Ocotea species is the 3-O-glycoside flavonol (C3)isoquercitrin, which constitutes a representative example, which has already been isolated from O. notata, O. elegans, O. corymbosa, and O. caudata. Isoquercitrin has been shown to possess antibacterial activity against Escherichia coli cells, inducing apoptosis-like death and damaging membrane dynamics by inducing oxidative stress. This underscores the potential bioactivity of Ocotea flavonoids, which warrants further exploration of their biological properties. Flavonoids have also been increasingly recognized for their antioxidant and pro-oxidant properties, as well as for relevant antiviral and antiprotozoal effects. , Flavonols can be used to seek more effective treatments for NTDs, such as leishmaniasis and trypanosomiasis. In addition, Ocotea species such as O. minarum and O. odorifera have been recognized as potent natural antioxidants. , Studies have reported antiviral effects against the herpes virus (HSV) and antibacterial effects for flavonoids of the O. notata leaves extract. , Recently, bacteriostatic effects were evidenced for O. minarum flavonoid fractions against the Salmonella, Pseudomonas, and Proteus genera. Furthermore, biflavonoids (C10) such as those found in O. odorifera and O. canaliculata, might be able to reduce the viability of tumor cell lines, as previously reported.

Besides, it is important to highlight that flavonoids, as other metabolites containing catechol units, are on the list of compounds with potential assay interference (PAINS). This is because they can provide promiscuous behaviors on screening assays since catechols can chelate metals as well as being reactive in the oxidized form to nucleophiles present in the side chains of proteins, interfering with assays or providing nonspecific activities. Thus, interpretation of the biological activities of flavonoids and their derivatives requires scrutinous analysis based on investigation of specific mechanisms of action.

A broad range of specific flavonoid glycoside derivatives was isolated from several Ocotea spp., such as O. vellosiana, O. odorifera, O. porosa, O. notata, O. lancifolia, and others. , Figures and present all the different flavonoid cores encountered in the genus, including flavones (C1), flavonols (3-hydroxyflavone or flavonol, C2), 3-O-glycoside flavonols (C3), 7-O-glycoside flavone-flavonols (C4), and flavanonols (C5). In addition, the 3-O-glycoside flavanonols (C6), flavan-3-ols (C7.1), phenylpropanoid-substituted flavan-3-ols (C7.2), 7-O-glycoside flavanones (C8), 8-glycoside flavones (C9.1), 6-glycoside flavones (C9.2), flavanonol dimers or biflavonoids (C10), tetrahydroxychalcones (C11), and flavonolignans (C12) are included.

18.

Chemical diversity of flavonoids in the Ocotea genus is expressed as the number of different chemical structures reported in the literature.

19.

Flavonoid cores found in Ocotea species: (C1) flavone, (C2) flavonol, (C3) 3-O-glycoside flavonol, (C4) 7-O-glycoside flavonol, (C5) flavanonol, (C6) 3-O-glycoside flavanonol, (C7.1) flavan-3-ol, (C7.2) phenylpropanoid-substituted flavan-3-ol, (C8) 7-O-glycoside flavanone, (C9.1) 8-glycoside flavone, (C9.2) 6-glycoside flavone, (C10) flavanonol dimer or biflavonoid, (C11*) tetrahydroxychalcone, and (C12 ^#) flavonolignan. *Flavonoid precursor; ^#flavonoid mixed biosynthetic class.

Ocotea spp. Terpenoids

Terpenoids represent the largest classes of chemicals found in the essential oils of Ocotea spp., followed by phenolic compounds, such as phenylpropanoid and phenylpropene derivatives, along with aldehydes, alcohols, and esters. Indeed, volatile terpenoids are the predominant constituents in the essential oils of a vast array of plants and flowers globally. − These compounds are biosynthesized via two pathways in superior plants: the mevalonic acid (MVA) and 2-C-methyl-D-erythritol-4-phosphate (MEP) pathways. The Ocotea species produce majoritarian sesquiterpenes via MVA and monoterpenes via MEP, indicating by our research that both pathways are highly evolved and active within the Ocotea metabolome (Supplementary Tables S5, S6, and S5.1–S6.1).

The isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) are the primary building blocks for terpenoid formation, and assemble in a head-to-tail fashion catalyzed by geranyl pyrophosphate synthase (GPP) to form the chemical core of most monoterpenoids (C₁₀), including alyphatic such as geraniol (22), citronellol (23), and linalool (24), and cyclic such as camphor (25), borneol (26), limonene (27), α-terpineol (28), and 1,8-cineole (29), which are all widespread in the Ocotea genus (Figure ). ,,,,−

20.

Biosynthesis of geranyl pyrophosphate synthase (GPP), common precursor of monoterpenes including geraniol (22), citronellol (23), linalool (24), camphor (25), borneol (26), limonene (27), α-terpineol (28) and 1,8-cineole (eucalyptol) (29).

The MVA pathway predominantly leads to the biosynthesis of sesquiterpenes (C₁₅), a major class of terpenes formed by the union of three isoprene units. These units originate from the farnesyl diphosphate (FPP) precursor, which is synthesized by the condensation of an IPP unit with GPP. The addition of the nucleophilic IPP unit to a GPP-derived allylic cation culminates in the formation of the C₁₅ farnesyl scaffold. The chain length can be increased by further modifications through oxidative cyclization reactions and rearrangements, giving rise to the largest chemical diversification among terpenoids. Thus, besides the significant number of linear structures, a vast range of sesquiterpene motifs, such as mono-, bi-, and tricyclic structures, can be biosynthesized. , The sesquiterpenes (+)-germacrenes A (30) and B (31) are sesquiterpenes frequently encountered in different Ocotea species essential oils (Figure ). ,

21.

Biosynthesis of FPP, the chemical core for C₁₅ derivatives, and sesquiterpenes (+)-germacrene A (30) and (+)-germacrene B (31).

Diterpenes (C₂₀) arise from the head-to-tail condensation of four isoprene units and are biosynthesized from the precursor geranylgeranyl diphosphate (GGPP), which is formed by the sequential addition of one IPP unit to FPP. Additional oxidative modifications catalyzed mainly by terpene synthases and cytochrome P450 enzymes, generate a broad diversity of diterpene skeletons. However, in Ocotea, diterpenes are comparatively uncommon metabolites and have been reported only in a few species, such as O. floribunda, O. neesiana, O. nutans, and O. bicolor (Supplementary Tables S5, S6, and S5.1–S6.1). Owing to this limited occurrence, diterpenes are unlikely to represent primary chemotaxonomic markers for the Ocotea species. Instead, they may still provide specific chemical profiles useful for the differentiation of species within the genus.

Terpenoid Profile: Monoterpenoids

Ocotea species are known for producing a diverse array of oxygenated and nonoxygenated, linear and cyclic monoterpenes. These compounds are distinguished by their varied chemical structures and unique biological activities, demonstrating a range of pharmacological properties, such as antimicrobial, antifungal, antitumor, and anti-inflammatory effects. Studies have suggested that the therapeutic potential of monoterpenes is due to their ability to interact with various biological targets, including enzymes, receptors, and ion channels. Such interactions can significantly affect cellular processes by modulating signaling pathways and influencing inflammation, apoptosis, and cell growth. ,,−

Most phytochemical studies on Ocotea report several monoterpenes without enantiomeric assignment, as no attempt to assign the absolute configuration of monoterpenoids has been made in the majority of essential oils chemical investigations reported. Among these, compounds 24, 26–29, β-myrcene (32), 4-terpineol (33), cis-β-ocimene (34), trans-ocimene (35), α-pinene (36), β-pinene (37), and camphene (38) are prevalent across various Ocotea species (Figure ). Specifically, monoterpene 32 is not only a major constituent in several plants but is also abundantly found in 22 Ocotea species (Tables S6 and S6.1). This compound is a highly important raw material for the production of flavors, fragrances, cosmetics, and also plays an important role in the pharmaceutical field, given its significant biological properties (e.g., anxiolytic, antioxidant, antiaging, anti-inflammatory, and analgesic). Similarly, 29 (also known as eucalyptol) has been found in 21 Ocotea species and is renowned for its anti-inflammatory effects, primarily through the inhibition of interleukins (IL) and interference with tumor necrosis factor alpha (TNF-α) production. It also exhibits antinociceptive, cardiovascular, and vasorelaxant properties. ,

22.

Widespread monoterpenes found in the Ocotea genus, including linalool (24), borneol (26), limonene (27), α-terpineol (28), 1,8-cineole (29), β-myrcene (32), 4-terpineol (33), cis-β-ocimene (34), trans-ocimene (35), α-pinene (36), β-pinene (37), and camphene (38).

Moreover, 29 and 33, which are commonly found in the tea tree (Melaleuca alternifolia), have been identified in 15 Ocotea species. These compounds act as anti-inflammatory agents by inhibiting the production of LPS-induced mediators such as IL-1β, IL-6, and IL-10. 29 is also utilized industrially as an ingredient in aromatic scents, perfumes, and cosmetics, given its lilac-like odor. Additionally, it exhibits a wide range of biological properties, including antioxidant, anticonvulsant, antiulcer, antihypertensive, antinociceptive, and insecticidal effects. Figure depicts the most commonly identified monoterpenes in the Ocotea genus.

Monoterpene linalool (24) has been found in 13 Ocotea species and is recognized as a biologically active compound of commercial interest due to its beneficial aromatic properties. This compound has demonstrated repellent properties against various crop-destroying insects and potential antioxidant, anti-inflammatory, anticancer, anxiolytic, analgesic, and sedative effects. , Moreover, monoterpene borneol (26) is a common component in plant essential oils, which is present in numerous medicinal plants, such as Valeriana officinalis, Matricaria chamomilla, and Lavandula officinalis; further, it is also present in 12 Ocotea species. In addition, 26 can be used in fragrances and cosmetics and exhibits potential for medicinal applications, pending reports of its biological activities, including its antimicrobial, anti-inflammatory, and antiviral effects. ,,

Terpenoids from several Ocotea species have shown promising antitumor properties against a variety of cancer cell lines, including breast adenocarcinoma epithelial cells (MCF-7 and MDA-MB-231), cerebral glioblastoma (A-172, U-87MG), cerebral astrocytoma (CCF-STTG1), and hepatocellular carcinoma (Hep-G2) cells. ,− Moreover, monoterpene α-terpineol (28), found in species such as O. quixos, O. bofo, and O. opifera among others, exhibit potent cytotoxic and antitumor effects, mediated through various mechanisms, including the induction of apoptosis, the inhibition of cell migration, DNA fragmentation, and cell cycle arrest.

Terpenoid Profile: Sesquiterpenoids

Sesquiterpenoids constitute the predominant terpenoid subclasses in the Ocotea genus and are major components of the essential oils extracted from various Ocotea species. In addition, several terpenoids can also be isolated from different parts of the Ocotea species, including barks, leaves, and flowers. ,,, Thus, the most prevalent compounds are presented in Figure and Supplementary Tables S5–6 and S5.1–61, including spathulenol (39), α-cadinol (40), elemol (41), bicyclogermacrene (42), caryophyllene oxide (43), globulol (44), α-humulene (45), β-caryophyllene (46), 1-epi-cubenol (47), viridiflorol (48), trans-nerolidol (49), and α-cadinene (50).

23.

Widespread sesquiterpenes found in the Ocotea genus, evidencing spathulenol (39), α-cadinol (40), elemol (41), bicyclogermacrene (42), caryophyllene oxide (43), globulol (44), α-humulene (45), β-caryophyllene (46), 1-epi-cubenol (47), viridiflorol (48), trans-nerolidol (49), and α-cadinene (50).

Spathulenol (39) was identified in 25 Ocotea species (e.g., O. diospyrifolia, O. lancifolia, and O. nutans). There are several aromatic medicinal plants in which 39 is recognized as a major volatile constituent, such as Psidium guineense, which is also a plant with ethnopharmacological significance and popularly used for treating a range of inflammatory diseases, as well as O. odorifera. In addition to its anti-inflammatory properties, 39 has also exhibited antioxidant, antiproliferative, immunomodulatory, and antibacterial effects. −

Likewise, α-cadinol (40) has been found in 24 Ocotea species, alongside other less concentrated isomers. Furthermore, the sesquiterpene 40 and its isomers can be encountered in the essential oils of other plant genera and are recognized for their antioxidant, cytotoxic, and antimicrobial activities. , For example, the sesquiterpene 40 and the diterpene 16-methylesteritol A displayed antibacterial potential against Xanthomonas with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 50 and 250 μg mL^‑1, respectively. Sesquiterpenes elemol (41), caryophyllene oxide (43), globulol (44), β-caryophyllene (46), and trans-nerolidol (49) have been identified in different isomeric forms and belong to the chemical composition of several Ocotea species (Supplementary Tables S6 and S6.1). Sesquiterpene viridiflorol (48) has been found in 11 Ocotea plant species and is noted for its anti-inflammatory action against leukocyte migration and antioxidant and antibacterial activities.

Ocotea spp. Essential Oils

Essential oils are complex mixtures of volatile organic compounds primarily biosynthesized in specialized plant cells. ,, Lauraceae species are highly valued for the essential oils produced by their plants, including those from Ocotea. Despite numerous studies, only 48 of 115 chemically characterized species (∼42.5%) have been examined in terms of their essential oil composition. To date, GC-MS analysis has allowed the identification of 491 different compounds as constituents of Ocotea essential oils, including a range of different monoterpenoids (Supplementary Tables S5–6 and S5.1–6.1). These compounds are present in conventional medicine and aromatherapy and are prevalent in cosmetics, dentistry, agriculture, food flavorings, cleaning products, and solvents, making them one of the most successful commodities in industry, ,,,, exhibiting biological potentials such as antimicrobial, antioxidant, anti-inflammatory, and cytotoxic effects. ,,,,−

Phytochemically, terpenoids represent the most abundant components in Ocotea spp. essential oils, with sesquiterpenoids (64.6%) being the dominant subclass, followed by monoterpenoids (28.5%). The qualitative and quantitative variations in these components can be influenced by factors such as genetic specificity, developmental stages, environmental stress, and adaptations to local climatic and soil conditions, as well as interspecies interactions. The essential oils of various Ocotea species also contain minor components such as aldehydes (1.8%), phenylpropenes (1.4%), phenylpropanoids (1.0%), alcohols (0.7%), ketones (0.6%), diterpenes (0.5%), and smaller percentages of aliphatic and aromatic hydrocarbons, carboxylic acids, and esters (Supplementary Tables S6 and S6.1).

Ocotea spp. Phenylpropanoid Profile

Phenylpropanoids constitute a diverse class of compounds commonly found in plant essential oils, primarily derived from the aromatic amino acid phenylalanine. Safrole, a well-known phenylpropanoid, is notably prevalent across the Lauraceae and was once suggested by Gottlieb as a chemotaxonomic marker for this family due to its widespread occurrence in several species. Interestingly, to date, the biosynthesis of safrole has only been confirmed in a few Ocotea species, such as O. pretiosa, O. odorifera, O. cymbarum, O. opifera, and O. zahamenensis. ,, Notably, safrole is the major component in O. odorifera essential oil, where its content can range broadly from 30% to 90%. Despite its prevalence, based on the current data available since Gottlieb’s initial studies, our review suggests that safrole may not serve as a reliable chemotaxonomic marker within the Ocotea genus. Nonetheless, it may still hold validity as a marker for other genera within the Lauraceae. This underscores the complexity and variability of the biosynthesis of phenylpropanoids and their potential utility in the chemotaxonomic classification of related plant groups.

Quantitative Chemical and Pharmacological Overview

The OcoteaDB included 984 metabolites reported from Ocotea species, spanning 115 species across the genus. These compounds are classified into major chemical groups, including terpenoids (n = 565), of which 491 were identified in essential oils, and 37 were isolated from crude extracts. However, this number of volatile compounds can be considered as approximate due to conventional GC-MS analyses limitations. Followed by lignoids (n = 182), alkaloids (n = 156), flavonoids (n = 61), and other compound classes (n = 57). Regarding species coverage, alkaloids (60 species), lignoids (45 species), flavonoids (20 species), terpenoids (15 species), and other classes (23 species) were reported. A graphical summary of this data is shown (Figure ), and a full breakdown by species is available in the Supporting Information (Tables S1–9). However, for the evolution of OcoteaDB as any genus-tailored chemical database, it should be integrated with geo-referenced data and tissue-specific profiles as more standardized reporting becomes available in the literature.

24.

Overview of isolated metabolites (left) and species (right) distribution in Ocotea spp. based on data compiled in OcoteaDB. *Essential oils included GC-MS reports of mono- and sesquiterpenes.

The biological activity of Ocotea metabolites reflects clear structure–activity relationships (SAR) among its major chemical scaffolds. Aporphine alkaloids, the most prevalent class in Ocotea, exhibit anti-inflammatory and cytotoxic activities, as shown by compounds like boldine, glaucine, and dicentrine, which act through mechanisms such as COX inhibition, NF-κB suppression, and TRPA1 activation. Benzylisoquinolines observed in O. odorifera, O. lancifolia, and O. caudata, such as 2, 3 and 4, have demonstrated anti-inflammatory, antiedematogenic, and anti-HIV effects, attributed to modulation of COX/LOX pathways, butyrylcholinesterase inhibition, and cytokine activity. ,,,,,,, In parallel, lignans and neolignans such as yangambin, licarin A, and verrucosin, predominantly found in species like O. cymosa, O. duckei, and O. macrophylla, are linked to antiprotozoal, larvicidal, and cytotoxic effects via disruption of protozoal viability, larval development, and mitochondrial integrity, supporting their relevance for neglected tropical diseases. ,,, These correlations between scaffold and dominant bioactivity highlight Ocotea as a pharmacologically important genus.

Recent Advances

Recent research has not only highlighted the cytotoxic and antitumor potential of the metabolome of Ocotea species but also demonstrated the significant role that Ocotea essential oils play in enhancing the efficacy of antibiotics against multidrug-resistant bacteria. The essential oil extracted from O. odorifera and its primary component, safrole, can enhance the activity of macrolide and aminoglycoside antibiotics, such as erythromycin and gentamicin, respectively. This is achieved through the direct inhibition of efflux pumps, which are crucial in modulating bacterial resistance. Consequently, the combination of O. odorifera essential oil and safrole with antibiotics leads to improved antibacterial activity and clinically relevant effects against Staphylococcus aureus. Furthermore, alongside their potential as therapeutic agents, recent nanotechnology applications combined with NP present a promising alternative for ecological management. This approach serves as a potential substitute for synthetic antiparasitic drugs and insecticides, which have been linked to significant environmental damage and adverse effects on nontarget organisms, including the development of insect resistance. ,, In this context, recently, the potential of a nanoemulsion containing essential oils from O. pulchella was evaluated as a control agent in the Schistosoma mansoni cycle. This nanoemulsion presented molluscicidal, ovicidal, and cercaricidal activities against the schistosomiasis transmitter Biomphalaria glabrata, causing the death of adults and preventing oviposition. The major component found in the studied essential oils is myristicin (methoxy-safrole −29.0%), followed by 36 (17.2%) and 42 (16.6%). Additionally, it showed relevant antiparasitic activity against S. mansoni, the infectious agent of schistosomiasis. Another research demonstrated that an optimized nanoemulsion containing essential oils from O. indecora leaves has been developed and tested for its larvicidal properties against A. aegypti larvae. The major constituent found in the essential oils was sesquirosefuran (81.4%). In silico analysis suggested that the larvicidal property is related to acetylcholinesterase enzyme inhibition. Notably, the nanoemulsion demonstrated no toxicity against the nontarget organism Apis mellifera, ensuring safety for pollinator bees. The formulation also showed excellent physical stability under both ambient and refrigerated storage conditions, highlighting its practical viability for vector control applications.

Conclusion and Perspectives

This review sheds light on the metabolome of the Ocotea genus, detailing the principal metabolic pathways that give rise to the bioactive scaffolds of alkaloids, lignoids, flavonoids, and terpenoids. The comprehensive overview of the chemical and biosynthetic characteristics provides a valuable resource for gaining relevant insights and guiding further research targeting Ocotea species. It synthesizes data from 115 different Ocotea spp. and 984 metabolites, including 156 alkaloids, 182 lignoids, 61 flavonoids, 528 terpenoids (including monoterpenes, sesquiterpenes, diterpenes, triterpenes isolated from crude extracts/identified in essential oils), and 57 metabolites from less-frequent classes. Ocotea species represent an important resource for sustainable bioprospecting, especially when studied within frameworks that value biodiversity protection and responsible use of native flora. Integrating chemical exploration with conservation awareness is especially pertinent for Lauraceae, many of which occur in ecologically sensitive or restricted habitats, including several Ocotea species. The presented chemical data are suitable for a scaffold library for drug discovery pipelines, in addition to aiding in species identification and chemophenetic studies, providing support for the integration of chemical data with taxonomical investigations in further evolutionary studies of the Ocotea genus and Lauraceae. Looking forward, the integration of OcoteaDB with emerging genomic and transcriptomic data offers a pathway to fully elucidate the biosynthetic gene clusters responsible for unique bioactive scaffolds present in the genus.

The review also discusses the specific stereochemical differentiation of alkaloids and the presence of common and uncommon NP classes among Ocotea species. The structural patterns and metabolites explored herein hold significant potential for the pharmaceutical and medicinal chemistry communities. Promising bioactive scaffolds were detailed, making the Ocotea species a valuable source for drug discovery. Future investigations are likely to reveal even more intricate biosynthetic pathways and novel compounds. OcoteaDB is an open-access community resource that can facilitate the use of these metabolites in high-throughput virtual screening, particularly in the search for novel anti-inflammatory and antiparasitic leads. Our analysis offers a comprehensive understanding of the Ocotea metabolome, providing valuable insights and laying the groundwork for continued advancements in several NP research fields related to the Ocotea plant species and its specialized metabolites.

The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.jnatprod.5c01567.

• Table S1: all Ocotea spp. from literature; Table S2: alkaloids; Table S2.1: alkaloids unique-species; Table S3: lignoids; Table S3.1: lignoids unique-species; Table S4: flavonoids; Table S4.1: flavonoids unique-species; Table S5: isolated terpenoids; Table S5.1: isolated terpenoids unique-species; Table S6: essential oils terpenoids; Table S6.1: essential oils terpenoids unique-species; and Table S7: other classes; Table S8: metabolites and Table S9: species; Table S1.1: Katchborian et al. 2024 - Ocotea NP classes hits positive mode; Table S1.2: Katchborian et al. 2024 - Ocotea NP classes hits negative mode (XLSX).

A.K.N. and D.A.C.P. contributed to the conceptualization and methodology of the manuscript. Both A.K.N. and K.J.N. were involved in all sections, including the methodology, writing, reviewing, and editing of the main sections of the manuscript. M.F.A., P.C.P.B., M.S.F., W.S.T., M.M.G., and D.O.S. were responsible for chemical data curation, reviewing, and editing. P.C.P.B., J.H.G.L., D.F.D., M.G.S., A.C.C.P., D.A.C.P., and R.E.E. provided resources, funding, and supervision and contributed in terms of critical thinking and to the writing, reviewing, and editing of the manuscript. All authors contributed to the final version of the review.

The authors express their gratitude to the funding agencies: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES Foundation, Brazil), finance code 001; Conselho Nacional de Desenvolvimento Científico e Tecnológico - Brazil (CNPq) (grant number 309500/2025-7, 304916/2025-0, 408229/2025-0, 408115/2023-8), Fundação de Amparo à Pesquisa do Estado de Minas Gerais - Brazil (FAPEMIG) (grant numbers APQ-02882-24, APQ-05607-24, APQ-05218-23, APQ-00544-23, BPD-00760-22), and São Paulo Research Foundation (FAPESP) - Brazil (grants number 2024/04606-5 and 2025/26427-8). The Article Processing Charge for the publication of this research was funded by the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), Brazil (ROR identifier: 00x0ma614).

The authors declare no competing financial interest.