Biomedicine & Pharmacotherapy 109 (2019) 1–20

Contents lists available at ScienceDirect

Biomedicine & Pharmacotherapy

journal homepage: [Link]/locate/biopha

Occurrence, chemical composition, biological activities and analytical T

methods on Copaifera genus—A review
Caroline Arrudaa, Jennyfer Andrea Aldana Mejíaa, Victor Pena Ribeiroa,
Carly Henrique Gambeta Borgesb, Carlos Henrique Gomes Martinsb,
⁎
Rodrigo Cássio Sola Venezianib, Sérgio Ricardo Ambrósiob, Jairo Kenupp Bastosa,
a
School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. Café s/n, Ribeirão Preto, SP, 14040-903, Brazil
b
University of Franca, Av. Dr. Armando Salles de Oliveira 201, Franca, 14404-600 Brazil

A R T I C LE I N FO A B S T R A C T

Keywords: Copaifera is a genus of large trees found in Brazil, mainly in Amazon forest, but also in Atlantic forest and cerrado
Copaifera biomes. It has also been found in other countries in South America. In Africa, it is found mainly in Congo,
Terpenoids Cameroon, Guinea and Angola. Its oleoresin has been used in folk medicine in the treatment of numerous healthy
Natural products disorders, such as urinary, respiratory, skin and inflammatory diseases, for which there are several studies
Pharmacological activities
corroborating its ethnopharmacological uses. It is also extensively employed in the pharmaceutical and cosmetic
Quality control
industries in the development of ointments, pills, soaps, perfumes, among others. Copaifera oleoresin contains
mainly diterpenes, such as: kaurenoic acid, kaurenol, copalic acid, agathic acid, hardwiickic acid, polyalthic
acid, and sesquiterpenes, comprising β-caryophyllene, caryophyllene oxide, α-copaene, α–humulene, γ-muur-
olene and β-bisabolol, among other compounds. On the other hand, Copaifera leaves contain mainly phenolic
compounds, such as flavonoids and methylated galloylquinic acid derivatives. Therefore, considering the eco-
nomic importance of Copaifera oleoresin, its ethnopharmacological uses, the need to develop new pharmaceu-
ticals for the treatment of many diseases, as well as the pharmacological potential of the compounds found in
Copaifera spp., it was undertaken a review covering mostly the last two decades on the distribution, chemistry,
pharmacology, quality control and safety of Copaifera species.

1. Introduction small, apetalous, hermaphrodites and arranged in axillary panicles, and

the fruits has ovoid seed surrounded by an abundant and colorful aril
Copaifera genus belongs to the family Leguminosae Juss., subfamily [1].
Caesalpinoideae Kunth, according to Engler system. Nonetheless, many There are divergent data on the exact number of species belonging
literature reports refer to these species as part of Caesalpiniaceae in to the Copaifera genus. According to the taxonomic database
accordance to Cronquist system. Classification only as Fabaceae is also International Plant Name Index ([Link]) [4], the genus has 106
found in several reports [1]. specific names and varieties and from these, 49 are considered valid,
Even when Léonard’s work [2] removed Copaifera from Guibourtia two names are misapplied, 49 are synonyms and six are unresolved. The
genus, and Dwyer [3] included African and American species in the high amount of botanical synonyms associated with some species re-
description, problems to delimitate the group still occurred because of inforce the need for more ecological, botanical, genomic, metabolomic
botanic complexity and morphological proximity to other genera. The and molecular detailed studies on the genus and a reassessment of the
distinction between Copaifera species is made mainly on the basis of species classification.
vegetative characters of the leaves, as presence or absence of scores on Regarding the folk names, plants belonging to Copaifera genus are
the leaflets, the number and shape of the leaflets, and floral char- known as “copaíba”, “copaibeira”, “copaífera”, “pau-de-óleo”, “palo-de-
acteristics, such as presence or absence of pubescent sepals, anthers’ bálsamo”, “aceite”, “copayer” and “cupa-yba”. In Venezuela, copaiba’s
lengths and pistil condition [3]. Briefly, these plants are botanical oleoresin is known as aceite de palo, cabimba, cabima, aceite de zaraza;
characterized by bearing petiolate and penultimate leaves. Flowers are in France as “huile de copahu”, “baume de copahu” or “huile rouge de

⁎
Corresponding author.
E-mail address: jkbastos@[Link] (J.K. Bastos).

[Link]
Received 13 July 2018; Received in revised form 3 October 2018; Accepted 9 October 2018
0753-3322/ © 2018 Elsevier Masson SAS. This is an open access article under the CC BY-NC-ND license ([Link]
 C. Arruda et al.

Table 1
Distribution of Copaifera genus.
Species Distribution

Argentina Bolivia Brazil Caribbean Colombia Costa Rica French Guiana Gabon Guyana Liberia Nicaragua Panama Paraguay Peru Suriname Venezuela Zambia

C. aromatica Dwyer X X X X
C. baumiana Harms X
C. brasiliensis Dwyer X
C. bulbotricha Rizzini & Heringer X
C. camibar Poveda, Zamora & Sanchez X X
C. canime Harms X X
C. cearensis Huber ex Ducke X
C. coriacea Mart X
C. depilis Dwyer X
C. duckei Dwyer X X
C. elliptica Mart. X
C. epunctata Amshoff X
C. glycycarpa Ducke X
C. grandifolia (Benth.) Malme X X
C. guyanensis Desf. X X X X X X X
C. gynohirsuta Dwyer X
C. jacquiniana G. Don X
C. jacquinii Desf. X
C. laevis Dwyer X

2
C. langsdorffii Desf. X X X X X
C. langsdorffii var. glabra (Vogel) Benth X
C. langsdorffii var. laxa (Hayne) Benth. X X
C. lucens Dwyer X
C. luetzelburgii Harms X
C. magnifolia Dwyer X
C. majorina Dwyer X
C. malmei Harms X
C. marginata Benth X
C. mildbraedii Harms X
C. multijuga Hayne X X
C. nana Rizzini X
C. oblongifolia Mart. X
C. officinalis L X X X X X
C. panamensis (Britton & Rose) Standl. X X X
C. paupera (Herzog) Dwyer X X X
C. piresii Ducke X
C. pubiflora Benth. X X X X
C. religiosa J. Leonard X
C. reticulata Ducke X X X X X
C. rondonii Hoehne X
C. sabulicola [Link] & L.P. Queiroz X
C. salikounda Heckel X
C. trapezifolia Hayne X X
C. utilissima Saldanha X
C. venezuelana Harms & Pittier X
Biomedicine & Pharmacotherapy 109 (2019) 1–20
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

Fig. 1. Distribution of Copaifera species.

copayer” [1]. or biological studies of Copaifera oleoresin. As an attempt to go forward,

The origin of the name of these trees came from indigenous lan- Albuquerque et al. [13] published a review comprising especially
guage “tupi”: they were called “cupa-yba”, meaning the reservoir/de- leishmanicidal activity data and Tobouti et al. [14] and Diefenbach
posit tree, because of the trunk oleoresin. In Argentina and Paraguay et al. [15] reported the antimicrobial effect of Copaifera spp.
they called them “copaíva” or “copahu” and “cupay”, respectively By analyzing these previous published reviews, it was observed that
[1,5]. Copaibas are shrubs or trees of slow growth, which can grow up they covered only data about the oleoresins from a few well-known
to 40 m of height, with a rough dark trunk measuring up to 4 m in species, and did not consider other important species of Copaifera
diameter. These trees are important source of oleoresin and wood [1,6]. genus, considering that there are more than 40 Copaifera species in the
The oleoresin is a liquid with variable viscosity and color, ranging world. Besides, there are still several information gaps on Copaifera
from yellow to light brown, composed by a solid phase or resin with a review papers, such as quality control and safety aspects, which are of
non-volatile portion comprising mainly acid diterpenes, and a volatile utmost importance: there is a wide inter and intra species chemical
oil part consisting of a mixture of sesquiterpenes. This oleoresin is variation, adulterations by the addition of soybean oil or other cheap
widely used in folk medicine and the majority of the published studies oils, as well as the mixture of different Copaifera species oleoresins in
are related to its chemical composition and pharmacological activities the commercial products. Therefore, this product has not been stan-
[5,7,8]. dardized yet, leading to difficulties in the replicating the biological
Although significant differences on the chemical profile of the assays, as well the safety assurance of this product. Also, phytochemical
oleoresin are observed among species and individuals from the same and biological reports of other parts of the plant are scarce. Another
species, the diterpenes belonging to kaurane, clerodane and labdane important issue is that the two first published reviews [1,5] are in
skeleton types can be identified in all oleoresins. The chemical struc- Portuguese, making it less accessible to the research community. Fur-
tures of all compounds cited on the text are in supplemental files and thermore, updating of published information contributes significantly
are referred according to the number attributed to each compound. to natural products scientific researchers. Therefore, in this review we
Copalic acid (184), a labdane-type diterpene, stands out because it is discuss ethnopharmacology, chemical composition, biological activ-
found in most of the oleoresin commercial samples [5,9]. In the volatile ities, quality control and toxicity of the accepted Copaifera species, their
fraction, the non-oxygenated and oxygenated sesquiterpenes like β- products and isolated compounds.
caryophyllene (12), α-copaene (9), α-humulene (16) and caryophyllene
oxide (2) are considered the main compounds, and sometimes are used
as chemical markers [5,10]. 2. Distribution of Copaifera spp
Recently, some studies have focused on the promising compounds
detected in the aerial parts. In this regard, Copaifera langsdorffii L. has Copaifera spp. are spread not only in West Africa with 19 species in
been the most studied one. Phytochemical analysis of the hydroalco- Congo, Cameroon, Gabon, Zambia and other countries, but also along
holic extract of the leaves revealed the presence of flavonoid hetero- the tropical region of America, from the north of Argentina to Mexico.
sides, such as quercetin-3-O-alpha-L-rhamnopyranoside (quercitrin Sixteen species can be found in Brazil, from the Amazonian forest in the
(177), kaempferol-3-O-alpha-L-rhamnopyranoside (afzelin (178), gal- North until the Atlantic and Riparian forests of the Southern Region
loylquinic and methylated galloylquinic acids (224 to 236) that are [1,16]. The most abundant species are C. officinalis L., C. guyanensis
known for displaying several important biological activities [8,11,12]. Desf., C. reticulata Ducke, C. multijuga Hayne, C. langsdorffii Desf., C.
To the best of our knowledge, reviews on chemical composition and coriacea Mart. and C. cearensis Huber ex Ducke (Table 1, Fig. 1). Dis-
biological activities of Copaifera genus were previously published by tribution of Copaifera species are displayed in Table 1 according to
Veiga Jr. and Pinto [1], Pieri et al. [5] and Leandro et al. [7], all of geographic information from Tropicos ([Link]) [16]. Ac-
them focusing on the ethnopharmacology, chemical composition and/ cording to this data base, C. coriacea Mart and C. martii are synonyms.

3
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

3. Ethnopharmacology 4. Chemical composition: secondary metabolites

Copaifera species have been known long before Brazil colonization In Copaifera spp. oleoresin, terpenoids are the major compounds. Its
in the 16th century. Indigenous tribes, by observing animals rubering its volatile fraction contains mainly sesquiterpenes caryophyllene, copaene
wounds in Copaifera’s trees, realized that it has healing and anti-in- and humulene, and the non-volatile fraction contains mainly acid di-
flammatory properties. Hence, they started using Copaifera’s parts, terpenes, such as kaurenoic, copalic and polyalthic acid. In other parts
especially the oleoresin, for medicinal purposes [17,18]. Paraguay’s of Copaifera tree it can be found compounds from different classes, such
indigenous also used the oleoresin to treat rheumatism, dysentery, as phenolic compounds [24]. The chemical structures of some major
especially with gangrenous retreat. In Venezuela and Colombia, the tea compounds isolated from Copaifera spp. are in Fig. 2. Although different
of the seeds and bark has been used as anti-hemorrhoidal and laxative species of Copaifera spp. have similar chemical composition from a
agent. In the Amazon (Brazil), besides wound healing, such teas are qualitative point of view, there is considerable variation in the con-
used for pulmonary diseases and asthma. In Cameroon, Copaifera re- centration of most of these compounds inter and/or intra-species.
ligiosa J. Léonard oleoresin is employed in the treatment of syphilis and Therefore, we considered important to summarize the chemical com-
blennorrhea [1]. position of several species of Copaifera. We also show the compounds
Due to the medicinal effects of Copaifera spp., this genus was one of described for several Copaifera species in Table 2.
the first ones described by Portuguese colonizers: it was found in a Gelmini et al. [25] identified several terpenoids both in the volatile
letter of 1534 from Petrus Martius to the pope Leão X, a mention of an and in the resinous fraction of C. langsdorffii oleoresin: the essential oil
oil used by indigenous called “Copei” and also in a letter from José de was composed mainly by sesquiterpenes (96.36%), as already reported
Anchieta, a Jesuit, to the General Father, in 1560, where he mentioned by other researches, highlighting the compounds 13 and 82. In this
copaiba oil uses. Since then, the physicians and the population started study, they found that the nonvolatile fraction comprised mainly acid
employing it in the treatment of diseases, considering the lack of diterpenes, such as 184. The HRGC and HRGC-MS analysis of C.
medications at that time [1,5]. In Europe, after knowing about the langsdorffii oleoresin allowed the identification of nine diterpenes, from
pharmacological potential of Copaifera spp., in 1587, it was registered which 184 stood out as a reference compound [9], as well as 155
in the “Brazilian Descriptive Pact” the medicinal application of co- (44.3%) and 190 (8.2%) acids as major diterpenes, and 12 (32.8%) as
paíba’s oleoresin, and in 1677 this product was added to the British the main sesquiterpene [26].
Pharmacopoeia and finally to “The United States Pharmacopoeia” in C. langsdorffi is one of the most studied species: in the volatile
1820 [18,19]. Copaifera oleoresin has been extensively used, especially fraction of its fruits the sesquiterpenes 38 (29.8%) and 12 (14.8%) are
in Amazon region, where “copaíba” is well known by its population, the major compounds [27]. Phenolic compounds, including 129, 159
due to the abundance of this genus in this region; it is also the most used and 160, among others were also present in C. langsdorffii fruits [24].
in folk medicine. It can be easily found in popular markets [7]. In other Extraction of fruit shells with hexane using Sohxlet apparatus led to the
Brazilian states, such as Rio de Janeiro, São Paulo, Espírito Santo, isolation of 155, 164, 166 and 2 [28], whereas in the peel oil, it was
Bahia, Mato Grosso do Sul, Minas Gerais, Paraná, Piauí, Ceará and also found 37, 58 and 38 [27]. From unripe seeds of C. langsdorffii, it
others, the most abundant Copaifera species is C. langsdorffii, although was also isolated 131 [28]. Its extraction with ether in a Soxhlet ap-
other species can be occasionally found in Brazil, such as C. lucens, C. paratus furnished 250.1 mg of 131 for each gram of seed meal [29] The
oblongifolia and C. trapezifolia [1,7,20]. compounds 9, 10, 49, 2 and 27 were also found in the seeds of C.
The oleoresin of Copaifera spp. is well known in folk medicine as langsdorffi. Compound 49, 2, 9 and 10 were found in the volatile por-
wound healing, as well as anti-hemorrhoidal, purgative, antic- tion of the pericarps of C. langsdorffi, as well [30].
arcinogenic, anti-inflammatory, anti-microbial, antiviral, local anaes- In C. langsdorffii root wood oil the following compounds were
thetic, cytotoxic, insecticide and bactericidal agent. Besides, the identified: 2, the major one, 37 and kaurene 153. Besides, 167 (31.9%)
oleoresin was also employed for treating diseases that were lacking and 7 (30.5%) were reported in trunk bark essential oil [27]. Wood
effective medications, such as blennorrhea and gonorrhea. In the 18th ethanol-water extraction afforded 441.9 mg of flavonoids per gram of
century, the use of this product for urinary diseases was frequent. Its extract, and 54.8 mg of tannins [31]. Compounds 38 (25.2%) and 12
pharmacological properties are usually attributed to the sesquiterpenes (16.6%) were the major volatile compounds in C. langsdoffii leaves
and diterpenes from the oleoresin [1,5,7,18] (Tables 2 and 3). Since the [27]. Variation of daylight intensity and water accumulation during
18th century, considering the discovery and development of new and different seasons had a strong influence in the sesquiterpene levels in
effective drugs, the prescription and uses of Copaifera products de- the essential oil obtained from C. langsdorffii leaves: in the wet season
creased. However, in recent decades, with the appeal for using herbals, 19, 2 and 49 were abundant, whereas in the dry season 19 level was
the use of Copaifera products has started to increase again. Never- higher only if the sample harvesting was performed around 5:00 pm
theless, it still lacks enough evidence of pharmacological efficacy and [32]. Supercritical fluid and LC–MS analyses were also carried out for
safety. Copaifera oleoresin is an important product in the Brazilian in- extract, allowing the identification of other compounds in C. lansdorffii
dustry, and most of it comes from Amazonia state, where Copaifera leaves: 155, 177, 178, 180 and 179. The studies conducted by Motta
species are abundant. Usually, this product is exported to Europe, North et al. [12] and Nogueira et al. [33] confirmed the presence of glyco-
America or sent to Brazilian southeast states [1]. It is also used in the sylated flavonoids 177 and 178, as well as galloylquinic acids and their
cosmetic industry, because of its anti-inflammatory, bactericidal and methylated derivatives (224–236), which were isolated from the hy-
emollient properties [18]. Another use of Copaifera oleoresin is in the droalcoholic extract of C. langsdorffii leaves, by using different chro-
fuel industry, due to the high amounts of hydrocarbons, being em- matographic techniques, including high-speed counter-current chro-
ployed as a green source of fuel [21–23]. Because of several applica- matography, Sephadex LH-20 and preparative HPLC.
tions of Copaifera oleoresin and other parts of these trees in folk med- In C. lucens oleoresin it was found higher amounts of 164 (69.2%)
icine in the last decades, many studies have been undertaken in the and 184 (11.1%), as well as 12 (1.8%) [26].
isolation and identification of the organic compounds found in these A seasonal study by GC–MS and GC/FID of the chemical composi-
species, as well as in the assessment of its biological activities, and the tion of C. martii oleoresin, revealed the prevalence and constancy of 9
correlation between the biological effects and the isolated compounds and 22, with variations between 36.4%–51.2% and 13.7%–17.3%, re-
(Table 4). spectively. Other compounds were 28, 38 and 37 [34]. Although, this
data contrast with the one described by Santos et al. [26], since they
detected 21 (10.7%) and 188 (29%) as the main sesquiterpene and
diterpene compounds in the oleoresin of C. martii.

4
 Table 2
Chemical compounds found in Copaifera species.
C. Arruda et al.

Number Compound Number Compound Number Compound Number Compound Number Compound

1 Ledol 48 Sesquisabinene 95 Elemol 142 (Z)-3-Hexenyl acetate 189 ent-15,16-Epoxi-13(16),14-clerodadien-18-oic

(Chrolechinic acid)
2 Caryophyllene oxide 49 Spathulenol 96 Globulol 143 Limonene 190 Hardwickiic acid
3 Guaiol 50 Valencene 97 Nerolidol 144 Phenylacetaldehyde 191 Patagonic acid
4 Cedrol 51 α -Bulnesene 98 Palustrol 145 3-Hexenoic acid 192 8-Hydroxylabdanoic
5 Cadalene 52 Torreyol 99 Rosifoliol 146 Nonanal 193 3-Cleroden-15,18-dioic acid
6 α-Cadinol 53 α-Muurolol 100 Viridiflorene 147 3-Hexenyl butyrate 194 (13E)-ent-Labda-713-dien-15-oic acid
7 β-Bisabolol 54 β-Chamigrene 101 Viridiflorol 148 Bicycloelemene 196 3-Methyl-5-(2,2,6-trimethyl-6-hydroxy-1-
cyclohexyl)-pentanoic acid
8 α -Cubebene 55 β-Curcumene 102 α-Amorphene 149 Bicyclosesquiphellandrene 198 Pinipholic acid
9 α -Copaene 56 γ-Curcumene 103 α-Bisabolene oxide 150 Ionone 199 3-Hydroxy-14,15-dinorlabd-8(17)-en-13-one
10 β-Cubebene 57 γ-Gurjunene 104 α-Caryophynellol 151 Caryophylla-4(12),8(13)- dien-5-β-ol 200 Kolavenol
11 Longifolene 58 δ-Elemene 105 α-Ylanglene 152 Abieta-812-diene 201 Isopimaradiene
12 β-Caryophyllene 59 Cyperene 106 β-Copaene 153 Kaurene 202 Octadecadienoic acid
13 α –Bergamotene 60 α-Santalol 107 δ-Amorphene 154 Kaurenol 203 Biformene
14 β-Sesquiphellandrene 61 3-Hydroxy-benzenemethanol 108 Calamenene 155 Kaurenoic acid 204 Cembrene
15 Aromadendrene 62 Aromadendrene epoxide 109 δ-Guaiene 156 Epicatechin 205 Daniellic Acid
16 α -Humulene 63 Bicyclogermacrene 110 Germacrene A 157 nor-Calamenen-10-one 206 Dehydroabietic acid
17 α -Curcumene 64 Calarene epoxide 111 7-epi-α-Selinene 158 Catechin 207 Methyl esther
18 ɣ-Amorphene 65 Cedrene 112 7-epi-Sesquithujene 159 Benzoic acid 208 Abietic acid
19 Germacrene D 66 Hujopsene 113 Aristolene 160 Gallic acid 209 Elaidic acid
20 Germacrene B 67 Cuprenene 114 (E,E)-Methyl farnesoate 161 Isoquercitrin 210 Labd-7-en-15-Oic acid
21 β-Bisabolene 68 Curcumenol 115 Geranyl Linalool 162 Procyanidin 211 Labd-8(20)-ene-15,18-Dioic acid

5
22 δ-Cadinene 69 Diepicedrene-1-oxide 116 Camphor 163 Rutin 212 Labdane
23 α -Cadinene 70 Elixene 117 Eudesma-4(15), 7-dien-1-β-ol 164 Polyalthic acid 213 Manool
24 α -Selinene 71 Eudesma-4(14),11-diene 118 Guaia-6,9-diene 165 Methyl polyalthate 214 Pimaric acid
25 β-Vetivene 72 Geranyl isobutyrate 119 Junenol 166 Nivenolideo 215 Sclarene
26 α-Caryophyllenol 73 Humulene epoxy II 120 Methyl geranate 167 Kaurenal 216 Valencene
27 1-epi-Cubenol 74 Isolongifolene, 4,5,9,10- 121 p-Cymene 168 Naringin 217 Dimethyl Agathate
dehydro
28 Alloaromandredene 75 Longipinocarveol 122 α-Copaen-11-ol 169 Morin 218 Dimethyl Pinifolate
29 Cubenol 76 Methyl 9-octadecenoate 123 β-Gurjunene 170 Luteolin 219 Methyl Kolavenate
30 Gleenol 77 Mustakone 124 Salvial-4(14)-en-1-one 171 Kampferol 220 Methyl Eperuate
31 trans-Cadina-1(2),4-diene 78 Selina-3,7(11)diene 125 cis-Calamenen-10-ol 172 Campesterol 221 Methyl Copalate
32 Zingiberene 79 β-Guaiene 126 nor-Calamenen-10-one 173 Estigmaterol 222 Pauperol
33 α-Calacorene 80 Zizanone 127 14-Hydroxy-α-muurolene 174 Stigmasta-5,22(E)-3dien-3β-ol 223 (-)-3β-Acetoxylabdan-8(17)-13-dien-15-oic acid
34 α-Gurjunene 81 α-Elemene 128 Cinnamic acid 175 β-Sitosterol 224 3-O-(3-O-Methyl galloyl) quinic acid
35 α-Muurolene 82 α-Himachalene 129 O-Coumaric acid 176 Kaur-16-ene 225 3,4-di-O-(3-O-Methyl galloyl) quinic acid
36 β –Selinene 83 α-Longipinene 130 P-Coumaric acid 177 Quercitrin 226 3-O-(Galloyl)-4-O-(3-O-methyl galloyl) quinic acid
37 β-Elemene 84 α-Santalene 131 Coumarin 178 Afzelin 227 3,5-di-O-(3-O-Methyl galloyl) quinic acid
38 γ –Muurolene 85 α-Thujopsan-2-ol 132 14-Hydroxy-α-cadinene 179 Dimethylquercetin 228 3-O-(Galloyl)-5-O-(3-O-methyl galloyl) quinic acid
39 γ-Cadinene 86 β-Eudesmene 133 Carotol 180 Eupatorin 229 3-O-(3-O-Methyl galloyl)-5-O-(galloyl) quinic acid
40 β-Farnesene 87 β-Humulene 134 Selin-311-dien-6α-ol 181 Methyl 4-methoxycinnamate 230 4,5-di-O-(3-O-Methyl galloyl) quinic acid
41 ɣ-Bisabolene 88 γ-Elemene 135 Zonarene 182 ent-8(17)-Labden-15-oic (eperuic acid) 231 3-O-(3-O-Methyl galloyl)-4-O-(galloyl) quinic acid
42 α -Bisabolene 89 Patchoulene 136 2,4 Decadienal 183 (13S)-7Labden-15-oic (cativic acid) 232 3-O-(3-O-Methyl galloyl)-4-O-(galloyl) quinic acid
43 α -Guaiene 90 δ-Selinene 137 Butylhydroxytoluene 184 Copalic acid 233 4-O-(galloyl)-5-O-(3-O-Methyl galloyl) quinic acid
44 Cyclosativene 91 G-Eudesmol 138 Dimethyl ethyl succinate 185 3ß-Acetoxycopalic acid 234 3,4,5-tri-O-(3-O-Methyl galloyl) quinic acid
45 β-Santalene 92 14-Hydroxy-α-humulene 139 Squalene 186 3ß-Hydroxy-copalic acid 235 3,5-di-O- (galloyl)-4-O-(3-O-Methyl galloyl) quinic
acid
46 β-Eudesmol 93 Acetoxy-caryophyllene 140 (Z)-3-Hexenol 187 ent-Agathic acid 236 3,4,5-tri-O-Galloyl quinic acid
47 Naphthalene 94 Calarene 141 2,4-Heptadiene 188 3,13-Clerodadien-15-oic (Kolavenic 237 ent-8(17) Labden 15,18-dioic acid
acid)
Biomedicine & Pharmacotherapy 109 (2019) 1–20
 Table 3
Biological activities and chemical compounds of Copaifera oleoresin.
SPECIES CHEMICAL COMPOSITION OF Copaifera OLEORESIN BIOLOGICAL EFFECTS ETHNOPHARMACOLOGY
C. Arruda et al.

DESCRIBED IN
SCIENTIFIC
LITERATURE
DITERPENES SESQUITERPENES References Oleoresin Disease Reference

Copaifera cearensis (182); (183); (184); (188); (1); (2); (3); (4); (5); (6); (7); (8); (9); [1,19,35,36,117] Anti-inflammatory [36]
Huber ex Ducke (189); (190); (191); (192); (10); (11); (12); (13); (14); (15); Antiparasitic
(193); (194) (16); (17); (18); (19); (20); (21); (Leishmania) [26]
(22); (23); (24); (25); (26) Antiparasitic
(Trypanosma) [81]
Copaifera coriacea (155); (188) (9); (8); (12); (13); (15); (21); (22); [26,34] Antiparasitic Wound Healing [118]
Mart (24); (27); (28); (29); (30); (31); (32) (Leishmania) [26,78] Antiulcer
(Copaifera (33); (34); (35); (36); (37); (38); (39) Antiparasitic
martii Hayne) (Trypanosoma) [81,84]
Copaifera duckei (164); (182); (184); (190); (2); (5); (7); (8); (9); (12); (13); (14); [17,37,38,56,83,119,120] Anti-inflammatory [54] – –
Dwyer (193); (196); (155) (15); (16); (19); (20); (21); (22); Antiparasitic (S.
(23); (24); (34); (35); (36); (37); mansoni) [71]
(40); (41); (42); (43); (44); (45); Antiparasitic
(46); (47); (48); (49); (50); (51); (antihelmintic) [72]
(52); (53); (54); (55); (56); (57); (58) Antiparasitic
(Trypanosoma) [82,83]
Copaifera (164); (184); (187); (155); (2); (8); (9); (12); (13); (14); (16); [39]
glycycarpa (198) (21); (37); (36); (40); (42); (43);
Ducke (44); (59); (58); (57)
Copaifera (164); (184); (190); (155) (2); (9); (12); (13); (21); (36) [17] Sore throat [121,122]
guyanensis

6
Desf.
Copaifera (167); (153); (155); (176); (2); (8); (9); (11); (12); (13); (14); [9,10,25,26,27,57,81,88,89,97,107,120,123,124,125,126,127] Gastroprotective [90]
langsdorffii (184); (185); (186); (187); (15); (16); (19); (20); (22); (24); Wound Healing [59]
Desf. (190); (155); (199); (200); (28); (29); (34); (35); (36); (37); Anti-inflammatory [25]
(190); (201); (202); (203); (38); (39); (40); (43); (44); (45); Antiparasitic
(204); (205); (206); (207); (46); (49); (51); (52); (54); (57); (Leishmania) [26]
(208), (209); (210); (211); (59); (60); (61); (62); (63); (64); Antiparasitic
(212); (213); (214); (215); (65); (66); (67); (68); (69); (70); (Trypanosoma) [81]
(216) (71); (72); (73); (74); (75); (76);
(77); (78); (79); (80); (81); (82);
(83); (84); (85); (86); (87); (88);
(89); (90); (152)
Copaifera lucens (164); (184) (12) [26,81] Antiparasitic
Dwyer (Leishmania) [26]
Antiparasitic
(Trypanosoma) [81]
Copaifera multijuga (184); (185); (186); (190); (1); (2); (3); (4); (5); (6); (7); (8); (9); [9,10,17,26,36,42,54,64,67,68,81,91,103,110,112,128,129,130,131,132,133] Anti-inflammatory Antypiretic [118,134,135]
Hayne (198); (202); (207); (217); (10); (11); (12); (13); (14); (15); [36,54] Wound Healing
(218); (219); (220) (16); (17); (18); (19); (20); (21); Antiparasitic Antiulcer
(22); (23); (25); (28); (29); (31); (Leishmania) [26] Sore throat
(34); (35); (36); (37); (38); (39); Antiparasitic Antipyretic
(40); (42); (46); (49); (52); (53); (Trypanosoma) [81] Flu
(57); (58); (59); (63); (65); (78); Antitumor [85] Antinociceptive
(81); (84); (88); (91); (92); (93); Anti-
(94); (95); (96); (97); (98); (99); inflammatory
(100); (101); (102); (103); (104);
(105); (106); (107)
(155); (184); (190) N.I. [51]
(continued on next page)
Biomedicine & Pharmacotherapy 109 (2019) 1–20
 Table 3 (continued)

SPECIES CHEMICAL COMPOSITION OF Copaifera OLEORESIN BIOLOGICAL EFFECTS ETHNOPHARMACOLOGY

DESCRIBED IN
C. Arruda et al.

SCIENTIFIC
LITERATURE
DITERPENES SESQUITERPENES References Oleoresin Disease Reference

Copaifera
oblongifolia
Mart
Copaifera (155); (182); (183); (184); (2); (4); (5); (7); (8); (9); (10); (12); [23,50,56,81,136,137,138] Anti-inflammatory
officinalis L. (186); (187); (190); (194); (13); (15); (16); (19); (20); (21); [66,139]
(155); (198) (22); (23); (24); (28); (32); (35); Antiparasitic
(36); (37); (38); (39); (40); (42); (Leishmania) [26]
(43); (49); (52); (53); (58); ((59); Antiparasitic
(79); (82); (105); (106); (108); (109) (Trypanosoma) [81]
Copaifera paupera (155); (164); (184); (186); (2); (6); (8); (9); (10); (12); (13); [38,40,140] Antiparasitic
(Herzog) (187); (195); (196); (155); (16); (21); (22); (23); (28); (29); (Leishmania) [26]
Dwyer (198); (221); (222) (31); (39); (53); (96) Antiparasitic
(Trypanosoma) [81]
Copaifera piresii – (2); (9); (12); (13); (16); (22); (23); [41]
Ducke (24); (36); (37); (58); (110); (111)
Copaifera pubiflora – (2); (9); (12); (13); [41]
Benth. (16); (22); (23); (24); (36); (37);
(58); (110);
(111)
Copaifera religiosa – – – Antiparasitic (P.
[Link] falciparum) [73]
Copaifera reticulata (155); (164); (182); (183); (2); (4); (5); (6); (7); (8); (9); (11); [26,36,42,43,44,45,54,77,141] Anti-inflammatory Leukorrhea, [118,142]

7
Ducke (184); (185); (187); (188); (12); (13); (14); (15); (16); (17); [36,54,64] ulcerations
(189); (190); (196); (223); (18); (19); (20); (21); (22); (23); Antiparasitic (P. Against Black
(24); (25); (28); (34); (36); (37); falciparum) [74] magic
(38); (39); (40); (41); (42); (44); Antiparasitic (R.
(45); (46); (49); (51); (53); (54); microplus) [76,77]
(55); (58); (59); (63); (65); (73); Antiparasitic
(81); (84); (88); (94); (96); (111); (Leishmania) [26,78]
(112); (113); (153) Antiparasitic
(Trypanosoma) [56]
Biomedicine & Pharmacotherapy 109 (2019) 1–20
 C. Arruda et al.

Table 4
Biological activities and chemical compounds of other tissue parts of Copaifera.
SPECIES CHEMICAL COMPOSITION OF Copaifera: OTHER TISSUE PARTS BIOLOGICAL EFFECTS DESCRIBED IN
SCIENTIFIC LITERATURE
Part Compound Reference Extract

Copaifera langsdorffii Desf. Leaf • No volatile [8,12,33,87,98,106,143,144,145] Gastroprotective [8] Anti-inflammatory [54]
(155); (177); (178); (179); (180); (181); (224); (225); (226); (227); (228); (229); (230); (231); Anticancer [86,87]
(232); (233); (234); (235); (236)
• Volatile [8,27,30,56,146]
(2); (3); (8); (9); (10); (12); (13); (15); (16); (19); (20); (22); (23); (27); (28); (31); (33); (35); (36);
(37); (38); (39); (46); (49); (52); (53); (58); (59); (63); (73); (77); (88); (95); (97); (98); (100);
(101); (102); (106); (108); (114); (115); (116); (117); (118); (119); (120); (121); (122); (123);
(153); (154); (155)
Fruit • No volatile [24]
(128); (129); (130); (156); (157); (158); (159); (160); (161); (162); (163)
• Volatile [27]
(2); (3); (8); (9); (12); (13); (16); (20); (22); (35); (36); (37); (38); (39); (46); (52); (58); (88); (95);
(108); (116); (121); (123); (155)
Fruit peel (2); (8); (9); (12); (35); (37); (39); (58); (155); (164); (166) [27,28]
Pericarp (1); (2); (8); (9); (10); (12); (13); (15); (16); (19); (20); (22); (23); (28); (31); (33); (35); (36); (37); [30]
(38); (39); (44); (49); (52); (53); (58); (59); (63); (73); (77); (79); (88); (95); (105); (106); (117);
(118); (120); (122); (124); (125); (126); (127); (153); (167)
Seed (2); (9); (10); (12); (22); (29); (33); (35); (38); (39); (49); (52); (53); (73); (77); (95); (101); (114); [30]
(117); (119); (120); (122); (124); (125); (131); (153)

8
Branch (1); (2); (3); (8); (9); (10); (12); (13); (15); (16); (19); (20); (22); (23); (27); (31); (35); (37); (39); [30]
(49); (53); (52); (59); (58); (63); (73); (77); (79); (88); (95); (97); (101); (105); (106); (108);
(114); (117); (118); (119); (120); (122); (124); (127); (132); (153); (154); (167); (380)
Stem bark (155) [95]
Root wood (2); (36); (153) [27]
Root bark (2); (12); (16); (24); (35); (37); (38); (46); (78); (79); (133); (134) [27]
Trunk wood (2); (35); (38); (39); (52); (53); (153) [27]
Trunk bark (7); (12); (153) [27]
Copaifera malmei Harms Leaf (158); (160); (163); (168); (169); (170); (171); (177) [48]
Copaifera multijuga Hayne Leaf (8); (9); (12); (22); (24); (36); (38); (39); (59); (87); (106) [132]
Copaifera officinalis L. Leaf resin (8); (9); (12); (36); (38); (39); (87) [147]
Leaf (8); (9); (12); (13); (16); (19); (20); (21); (22); (23); (28); (31); (35); (37); (38); (39); (58); (102); [49]
(105); (108); (110); (112); (135)
Stems (9); (12); (16); (19); (20); (21); (22); (23); (28); (31); (33); (34); (35); (36); (37); (38); (39); (48); [49]
(58); (102); (105); (108); (112); (135)
Seeds (131); (136); (137); (138); (139); (172); (173); (174); (175) [148,149]
Roots (8); (9); (12); (16); (19); (21); (28); (35); (37); (39); (48); (58); (102); (105) [49]
Copaifera panamensis (Britton & Leaf resin (2); (9); (8); (12); (22); (24); (36); (39); (38); (59); (87); (106) [147]
Rose) Standl.
Copaifera reticulata Ducke Bark Profisetinidin type tannins [46]
Copaifera trapezifolia Hayne Leaf • Volatile compounds [47]
(2); (6); (8); (9); (10); (12); (15); (16); (19); (22); (23); (28); (31); (33); (34); (35); (37); (38); (39);
(44); (49); (62); (63); (67); (88); (97); (101); (102); (108); (123); (131); (140); (141); (142); (143);
(144); (145); (146); (147); (148); (149); (150); (151); (176)
Copaifera venezuelana Harms & Leaf resin (9); (8); (36); (38); (59); (87) [147]
Pittier
Biomedicine & Pharmacotherapy 109 (2019) 1–20
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

Fig. 2. Chemical structure of some major compounds found in Copaifera spp.

Phytochemical study of C. cearensis Huber ex Ducke by Braga et al. species.

[35] and Pinto et al. [19] led to the isolation of several sesquiterpenes Analyses of the infusion extract of the leaves of C. malmei revealed
and diterpenes, which were identified and quantified by Veiga et al. the presence of phytosterols and phenolics including flavonoids, such as
[36] by High Resolution Gas Chromatography coupled to a mass 163, 160, 209, 177 and 158 [48].
spectrometer. They found that 12 (19.7%) as the major sesquiterpene Leaves, stems and roots volatile compounds of C. officinalis were
and 189 (11.3%) as the major diterpene in this sample. Additionally, it also studied to recognize the difference between three-week old seed-
was detected the compounds 1 and 2, which is responsible for the smell lings (after emergence) and two-year old trees. In seedling and two
of C. cearensis oil [1]. years old leaves and stems, the main compound was 19. Also, 12 was
It was identified in C. dukei Dwyer 12, 13, 37, 24 and 21 as major present in the leaves in the two stages of life (22.98% and 53.41%), and
sesquiterpenes, as well as the diterpenes 155, 184, 164, and 190 in all plant tissues. In the two years old leaves, increasing amounts of
[17,37,38]. The chemical profile of C. duckey changed along the year other sesquiterpenes were detectable, which was the case of 149 and
and among samples from different locations, revealing a large variation 39. In the roots, the amount of sesquiterpenes increased with the years,
in its composition [17]. with mainly the presence of 12, 105, 16 and 34 [49].
De Souza et al. [39] identified 30 compounds in the oleoresin of C. Chemical analyses of C. officinalis by CG-FID and CG-MS, revealed
glycycarpa Ducke and isolated the two major diterpenes 155 (0.98%) the presence of 12 (2632%) and 164 (1169%) as major compounds in
and 164 (19.73%). The most abundant sesquiterpenes were 12 the volatile and non-volatile portion, respectively [50]. Other studies
(18.53%) and 21 (8.25%). stated 190 and 184 as the main diterpenes [26]. In C. oblongifolia, the
The most abundant volatile compound in C. guyanensis Desf is 2 same diterpenes were found in higher amounts [51].
(19.1%) and the main diterpenes are 155 (17.5%), 190 (11%) and 164 In the oleoresin of C. venezuelana the main identified compounds by
(10.6%) [17]. In C. paupera (Herzog) Dwyer, besides several known IR and CG/MS were 59 and 9. Further analyses using polar and non-
sesquiterpenes and diterpenes, it contains the novel diterpene 222. The polar CG columns confirmed the presence of other classes of secondary
diterpene 184 is the most abundant compound in the non-volatile metabolites, commonly found in the genus [52]. The volatiles com-
fraction of the oleoresin, while 9 comprises 42.5% of the volatile pounds in leaf of C. panamensis, obtained by steam-distillation, 24 and
fraction [40,41]. 36(65%), 2 (11.5%), and 59 (6.7%) were listed as the compounds
In C. piresii Ducke and C. pubiflora Benth., the predominant sesqui- found in higher amounts [52].
terpenes are 9 (45.5%) and 12 (65.9%) [41]. The different species of Copaifera usually have a complex chemical
The major reported volatile compound in C. reticulata Ducke is 12 composition regarding the oleoresin and other parts of the plant; and
[26,42,43], although Badarjí et al. [44] reported 21 as the major several factors can influence the presence and amount of each com-
compound (24.91%) followed by 13 (21.99%). The major diterpene pound in the samples, such as genetics of each species, climate and
compounds are 190 (0.0849 g/L), 184 (0.0810 g/L) and 155 (3.9%) harvesting conditions. One other issue that frequently occurs that af-
[36,42]. Santos et al. [26] and Zoghbi et al. [45] analyzed samples fects the chemical composition is adulterations. Therefore, considering
collected from different locations and found a wide variation in the these aspects of Copaifera chemistry, validated analytical methods for
amounts of both sesquiterpenes and diterpenes in the oleoresin. In the the quality control of these samples are important to both quantitative
bark methanolic extract of C. reticulata, it was detected profisetinidin and qualitative point of view, because there is a relation between
tannin type [46]. chemical constitution and biological properties reported.
In C. trapezifolia Hayne, 12 (33.5%) was described as the major The chemical composition associated with corroborated biological
volatile compound in the leaves by Veiga et al. [47] who reported this effects and ethnopharmacology are shown in Tables 3 (oleoresin) and 4
compound in high amounts also in the oleoresins from other Copaifera (other Copaifera parts). The chemical structures of the compounds from

9
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

the table are in supplemental files. extract, their isolated diterpenes and sesquiterpenes may display bio-
logical effects with pharmacological importance, several reports have
5. Biological activities evaluated their efficacy against many human pathologies, such as in-
flammatory diseases, cancer, bacterial, fungal and parasitic infections,
By comparison between the papers that describe phytochemical among others [7].
studies and the ones that describe biological activities of Copaifera spp. Several diseases do not have available medications in the market
oleoresin, it was observed that, because of the presence of sesqui- that are totally effective and without important side effects [58].
terpenes and diterpenes in this product, several biological effects have Therefore, novel drugs should be developed for treating these and
been described: among the volatile constituents, compound 12 stands several other illnesses. In this field, natural products have an important
out, since it is found in practically all the analyzed species. It has been place, since they are used as healing agents since the beginning of ci-
reported that the presence of this compound in the oleoresin is related vilizations [26]. In this regard, Copaifera oleoresin and extracts from
to the anti-inflammatory property of the oleoresin Veiga et al. [36]: due other plant tissues have been evaluated in the past few decades against
to its presence in the essential oil of C. multijuga, it presents anti-in- numerous pathologies and parasites. The biological effects of Copaifera
flammatory activity by inhibition of some inflammatory mediators spp., and the chemical composition are displayed in Tables 2 and 3,
[53]. comprising the oleoresin and other plant parts, respectively.
Regarding the non-volatile compounds, they seem to be related to
several biological activities; for example, compound 155 is associated 5.1. Wound healing
with the anti-inflammatory activity of C. langsdorffi [54]. It has also
been reported the antiparasitic activity of diterpenes found in Copaifera One of the most established traditional uses reported for Copaifera
spp, such as 164 (IC50 of 3.87 μg/mL) that has a trypanocidal effect oleoresin is as a wound healing agent. Despite of this, there are few
against Trypanosoma brucei [55] and hydroxycopalic acid (IC50 of scientific reports corroborating the effectiveness of Copaifera spp. re-
2.5 μg/mL), with leishimanicidal activity [56]. There are also reports garding this activity. Taking it into account, Paiva et al. [59] evaluated
showing that the oleoresin diterpenes display antimicrobial effect: 184 the C. langsdorffii oleoresin in vivo: daily topical application of 100 μL of
(MIC between 5 and 0.5 μg/mL) inhibited the growth of several mi- solutions with 2% or 4% of the oleoresin were carried out during 21
croorganisms such as K. rhizophila, S. pyogenes, S. pneumonie, B. subtillis, days. It was found that at 4%, the oleoresin accelerated significantly the
S. dysgalactiae, S. agalactiae, S. epidermidis [57]. Kaurenoic acid (155) healing process: 84.05% of reduction of the original size, in comparison
displayed similar effect on S. pneumoniae and S. pyogenes at 5 μg/mL with 51.29% of control on the 9th day. Tensile strength in healing in-
[57]. The oleoresin of C. reticulata inhibited the growth of P. gingivalis at cised also showed a difference when they were treated with 4% of
6.25 μg/mL [44]: this bacteria is part of the oral bacteria responsible for oleoresin at day 5 (71.48 g/cm on the oleoresin treatment, in compar-
causing several diseases and a novel antimicrobial natural agent is ison with 35.95 g/cm on the control).
important considering the actual resistance of various bacteria strains Treatment with copaiba oleoresin itself and oleoresin cream based
[58]. (10%) were evaluated topically on rabbit’s wounds, finding out that
Besides, according to TCMSP data base ([Link] both treatments promoted significant wound healing, reassuring its
[Link]), some of the major compounds found in Copaifera spp. pharmacological potential [60].
display significant biological effects by acting in specific targets. For Despite the promising effects on the use of Copaifera oleoresin as
example, compound 155 is a potential candidate for the development of healing agent, the lack of a positive control makes it difficult to eval-
drugs for the treatment some important disease, such as: breast cancer, uate the real potential of Copaifera as a wound healing natural product.
by interacting with progesterone receptor; hypertension, by acting on Besides the few research studies covering Copaifera’s healing activity
mineralocorticoid receptor; Alzheimer; atherosclerosis; and lung and the lack of positive controls, the mode of action has not been ac-
cancer, by targeting the enzyme myeloperoxidase. Additionally, β-car- tually elucidated yet, although some authors reported increased vas-
yophyllene (12) interact with different target proteins, such as: pros- cularization and increased fibroblast production, which may contribute
taglandin G/H synthase 1, by displaying anti-inflammatory effect; to this effect [61]. Also, there is a lack of studies concerning different
muscarinic acetylcholine receptor M1, being useful in the treatment of copaiba species wound healing effect. They are relevant because of the
alzheimer’s disease, bronchospasm, cognitive deficits and schizo- chemical composition, which varies among species, and is directly as-
phrenia; with γ-aminobutyric-acid receptor α-2 subunit, acting as anti- sociated with biological effect [61]. In addition to the mode of action, it
anxiety and anti-insomnia agent; and with retinoic acid receptor RXR- is necessary to define whether the healing activity of the oleoresin is a
α, showing an anti-prostate cancer effect. α-Humulene (16) acts on result of the antimicrobial and immunomodulatory properties attrib-
prostaglandin G/H synthase 2, displaying a potential for treating ab- uted to some species of Copaifera or not. Hence, only after several ad-
dominal aortic aneurysm, Alzheimer's disease, pain, arthritis, prostate, ditional research studies approaching these and other pharmacological
bladder and breast cancer, endometriosis, inflammation and stroke. It information, the traditional use of Copaifera as healing agent will be
also interacts with the enzyme alcohol dehydrogenase, making it useful accordingly corroborated.
for treating alcoholism. Other targets of this compound are sodium-
dependent noradrenaline transporter and γ-aminobutyric acid receptor 5.2. Anti-inflammatory and antinociceptive
subunit α-1, which are related with depression and anxiety, respec-
tively. Among the medicinal plants used in folk medicine for the treatment
Although many compounds found in Copaifera spp. display sig- of inflammatory disorders, Copaifera spp. have showed a great potential
nificant effects, attributing a biological activity to a particular com- [62]. Both the anti-inflammatory and the antinociceptive activities of
pound present in a complex sample such as copaiba oleoresin is not an copaiba oleoresin are among the most well-studied biological activities
easy task, because in many cases it may be a synergic or additive effect of this plant: Veiga et al. [36] evaluated the anti-inflammatory activity
among the compounds present in the sample. An example is the gas- of the oleoresin from C. cearensis, C. reticulata and C. multijuga by
troprotection attributed to the hydroalcoholic extract of leaves of C. measuring the NO production in murine macrophages, and in vivo by
langsdorffi, which can be biologically active due to the presence of using the zymosan induced pleurisy model in mice. Diclofenac
compounds with this property [8]. Another example is the anti-in- (100 mg/kg) was used as reference inhibitor (positive control). It was
flammatory activity of 12 and 155, which can be found together in found that among the evaluated samples, the oleoresin of C. multijuga
several species of Copaifera, increasing the pharmacological effect [26]. significantly inhibited the NO production in vitro by 30% at 50 μg/mL.
Therefore, considering that the oleoresin, other Copaifera parts In vivo, the total leukocyte accumulation inhibition were 45% and the

10
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

neutrophil accumulation decreased by 73% after oral administration of antinociceptive effects, reducing writhing response and inflammatory
the oleoresin at 100 mg/kg. The oleoresin of C. cearensis also inhibited pain. Likewise, the hexanic, chloroformic and methanolic fractions of
approximately 30% of the NO production in vitro at 50 μg/mL, as well the oleoresin at doses ranging from 1 to 150 mg/Kg inhibited the
as total leukocyte, neutrophil accumulation and Evans blue extravasa- number of contortions induced by acetic acid and inflammatory pain.
tions at 100, 200 and 400 mg/kg in vivo. C. reticulata oleoresin also Doses of 50 and 100 mg/kg of the hexanic and chloroformic fractions
inhibited NO production, but in a higher concentration (85% at 500 μg/ displayed higher antinociceptive effect in comparison with morphine
mL), and total leukocyte and neutrophil accumulation at 400 mg/kg in (1 mg/Kg), indicating their action in opioid receptors, as previous de-
a similar response to diclofenac at 100 mg/kg. scribed by Gomes et al. [70]. All fractions inhibited the rat paw edema
C reticulata also promoted a reduction of the chronic inflammatory and increased vascular permeability at 150 mg/Kg [42,70].
infiltrate with a biological response similar to the positive control Although the anti-inflammatory and antinociceptive properties of
(dexamethasone) [63]. copaiba oleoresins are well established for some species such as C.
Thereby, considering the high amounts of compound 12 found in langsdorffi, other species still lack studies on this activity, mainly con-
these Copaifera species, may contribute significantly to the anti-in- cerning the determination of which compounds are responsible for the
flammatory activity of the oleoresin. activity. It is important to establish the molecular bases involved in the
Anti-edematogenic activity of the oleoresin from C. multijuga and its modes of action of these oleoresins against the inflammatory process
fractions were assessed in a rat paw’s edema model: dichloromethanic and nociception. Besides, to the knowledge of the authors, the phar-
and methanolic fractions, administered intraperitoneally at 30 mg/Kg macokinetic profile of the terpenoids from Copaifera has not been
caused a significant inhibition of paw edema produced by carrageenan evaluated. Therefore, no assumption regarding the absorption, bioa-
with inhibition of 49% and 64%, respectively, after 2 h. The hexanic vailability, metabolization and excretion of them can be done, although
fraction also significantly inhibited paw edema formation induced by some absorption of the compounds from the oleoresin and from dif-
bradykinin by 50% after 30 min [64]. ferent fractions such as dichlorometanic, methanolic and hexanic seem
Orally administered C. officinalis oleoresin displayed an expressive to occur since after oral administration, there is significant pharmaco-
inhibitory effect on paw edema induced by carrageenin and decreased logical effect in several cases. Thus, although its traditional use is pri-
new granuloma formations in a similar effect in comparison with marily characterized by anti-inflammatory activity, there are still many
phenylbutazone. It also displayed a 5-lipooxygenase inhibitory activity studies needed to fill the gaps that surround this subject, supporting its
in vitro [65,66]. popular use in a safe way for society.
Kobayashi et al. [67] observed that C. multijuga volatile fraction of
the oleoresin at 100 mg/kg and 200 μL/mL reduced in vitro the migra- 5.3. Antiparasitic
tion of leukocytes to pleural cavity of rats. Vargas et al. [68] performed
in vitro anti-inflammatory tests with the oleoresin isolated diterpenoids: The antiparasitic activity of Copaifera species have been evaluated
3-hydroxy-copalic (186) acid inhibited the enzyme tyrosinase (64%) at by several researches, considering the effects of the oleoresin, volatile
250 μM, while copalic acid (184) showed a significant NO inhibition fraction and isolated compounds, especially against parasites re-
(98.5%) at 100 mM, which was activated by lipopolysaccharide. 3- sponsible for causing neglected tropical diseases. There is a demand for
Acetoxy-copalic (185) and copalic acids (184) inhibited the production new drugs, since the drugs in the market can cause numerous serious
of IL-6 by 23.8% and 4.2%, respectively, at 25 μM and increased IL-10 side effects. Schistosomicidal assessment of C. duckei oleoresin and its
production. isolated compounds 164, 195, 182, 196 was reported by Borges et al.
Treatment of THP-1 stimulated monocytes with C. langsdorffi [71]. The authors found that the oleoresin displayed a promising LC50
oleoresin reduced the release of pro-inflammatory cytokines (IL-1β, IL- of 75.8 μg/mL after 24 h of incubation against Schistosoma mansoni and
6, TNFα) in a dose-range of 0.1–10 μM. Also, the oleoresin inhibited terpenoid 164 showed an LC50 of 41.7 after 24 h of incubation. Fur-
cytokine secretion, a consequence of the interaction with the NF-κB thermore, the oleoresin and 164 were able to modify the production
signaling pathway [25]. The isolated 155 also showed an important and development of eggs, and the first one affected the tegument of the
effect as anti-inflammatory NO inhibitor. This effect is extremely im- adult worms. It was also demonstrated a synergistic and/or additive
portant because the excessive production of NO is highly associated effect of the two main compounds of the oleoresin. By observation of
with serious inflammatory and autoimmune diseases. Diterpene 155 these IC50 values, the isolated compound 164 and the combination of
displayed IC50 = 61.2 ± 4.2 μM and inhibited the production of IL-6 164 with 237 in a proportion of 3:1 w/w showed considerable high IC50
by 11.2% at 50 μM. Although in one hand it increased the IL10 pro- concentrations (41.7 and 22.1, respectively) in comparison with the
duction, an anti-inflammatory interleukin, at 12.5 μM, on the other positive control praziquantel (IC50 of 0.269 μg/mL). On the other hand,
hand it decreased its production at 50 μM [68]. the selectivity index of 164 (3.07) demonstrated that this acid diterpene
C. langsdorffii hydroalcoholic leaves extract reduced edema forma- is selective to the parasite. Thus, regarding 164 effect against Schisto-
tion on two different models at doses of 100, 200 and 400 mg/Kg, with soma, it can be inferred that it is noticeably less potent than prazi-
a maximal inhibition of 52.3%. However, the study showed that this quantel, but it is selective to the parasite. Considering the oleoresin,
effect was not associated with cell migration inhibition, or cytokines even though the IC50 concentration (75.8μg/mL) is higher than prazi-
regulation [69]. Santiago et al. [54] reported that Copaifera oleoresins quantel, this natural product contains a complex mixture of several
(C. reticula, C. duckei and C. multijuga) were able to act on human compounds, especially sesquiterpenes and diterpenes. Therefore, it can
monocytes and cytokines, modulating the immunologic system, which be a promising herbal medicine candidate without the need of isolation
is extremely important due to cytokines such as TNF-α be responsible of active constituents. Nonetheless, before becoming a herbal medicine
for numerous immunologic diseases, and a low production of it may candidate, several other aspects should be considered: the standardi-
lead to unrestrained infections. The oleoresins were capable of inducing zation of the oleoresin chemical composition, the identification of the
LPS, preserve TNF-α levels and inhibiting IL-10 production. compounds responsible for the effect and analytical methods for per-
C. reticulata had its antinociceptive effect assessed in vivo furnishing forming its quality, which is directly related to the pharmacological
IC50 of 30 mg/kg on AA-induced writhing, 45 mg/kg on formalin model activity.
and 30 mg/kg on tail flick model, respectively. Regarding the mode of The oleoresin of C. duckei was also tested against monogeneans by
action, it suggests that it acts in the opioid receptors, therefore inducing Da Costa et al. [72], since the main products used to treat this disease
central and peripheral antinociceptive effect [42]. are toxic both to fishes and to the environment. The antihelmintic effect
C. multijuga Hayne also display antinociceptive effect: doses ranging of the sample against Anacanthorus penilabiatus and Mymarothecium
from 30 to 150 mg/kg significantly displayed peripheral and central viatorum in vivo revealed a decrease of parasitaemia of approximately

11
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

45%, when the fishes were bathed with 50 mg/L of the oleoresin. promastigotes with IC50 values of 2.5 and 6.0 μg/mL, respectively.
Besides, C. religiosa J. Leonard was evaluated by Lekana-Douki et al. Compounds 198 and 155 were active against axenic amastigote and
[73]: the dichloromethanic extract was tested against FCB (Chlor- displayed IC50 values of 3.5 and 4.0 μg/mL, respectively. The ones 187,
oquine-resistant), 3D7 (chloroquine-sensitive) and field strains of 155 and 198 caused significant increases in plasma membrane per-
Plasmodium falciparum. They discovered that the sample demonstrates meability and mitochondrial membrane depolarization of the proto-
promising IC50 values (13.4 ± 3.6 μg/mL, 8.5 ± 4.7 μg/mL and zoan [56].
1.5 μg/mL). Soares et al. [79] assessed the leishmanicidal activity of Copaifera
De Souza et al. [74] performed in vitro and in vivo evaluation of C. spp. commercial oleoresins and found that sesquiterpene rich samples
reticulata oleoresin against Plasmodium falciparum W2 and 3D10 strains, demonstrated good activity with IC50 of 2.9 and 2.3 mg/mL (against
and erythrocytes infected by P. berguei, respectively. The pharmacolo- intracellular amastigote). Since β-caryophyllene (12) is the major ses-
gical potential of Copaifera species for malaria treatment can be re- quiterpene from several Copaifera oleoresins, they also tested the iso-
inforced considering its IC50 of 1.66 ± 0.046 μg/mL (W2) and lated compound: it showed an IC50 of 1.3 μg/mL, which corresponds to
2.54 ± 0.057 μg/ml (3D10) and the significant reduction of para- 6.4 μM. They also found that this biological activity is probably not
sitemia in mice at 100 and 200 mg/kg/daily. Considering the phar- related to nitric oxide production. Hence, this compound seems to have
macological potential of both C. religiosa and C. reticulata against P. an important role in the leishmanicidal activity of the oleoresins and
falciparum, since the IC50 of the samples were below 20 μg/mL [75], may be a potential drug candidate against Leishmania amazonensis.
these samples may be considered for further studies against this para- Although, in comparison to positive controls such as amphotericin
site aiming to develop a novel herbal product against malaria. B, the isolated sesquiterpenes and diterpenes, as well as Copaifera
C. reticulata oleoresin also displayed a significant effect in control- oleoresins are usually less active, they are considered promising drugs
ling Rhipicephalus (Boophilus) microplus, a parasite responsible for for development of novel medications against Leishmania parasites.
transmission of “Southern cattle tick”, showing an LC50 of 1579 ppm According to the literature, samples with IC50 below 2 μg/mL for ex-
and LC99 of 3491 ppm after 24 h treatment [76]. Moreover, the oleor- tracts are considered to present a good activity and with IC50 from 2 to
esin at 5% and 10% were able to modify Rhipicephalus microplus in vitro 20 μg/mL are considered to present moderate effect [75]. For pure
reproduction, since it diminishes the quantity of females to perform compounds the same values in μM are used. Hence, numerous species of
oviposition [77]. Copaifera and some of their major isolated compounds may be con-
sidered for further studies aiming at developing novel antiparasitic
5.3.1. Antileishmanial herbal medicines.
Several species of Copaifera have been assessed against Leishmania
species and Santos et al. [26] found that C. reticulata from Pará state- 5.3.2. Trypanocidal
Brazil, C. reticulata from Acre state- Brazil, C. martii, C. cearensis, C. Drug discovery of new agents against Trypanosoma, the parasite
paupera, C. langsdorffii, C. officinalis, C. multijuga and C. lucens displayed responsible for causing Chagas disease, is relevant since approximately
leishmanicidal activity against promastigote forms of Leishmania ama- 7 million people are already contaminated by Trypanosoma, and other
zonensis, with IC50 (μg/mL) of 5.0 ± 0.8, 22.0 ± 0.0, 14.0 ± 0.9, 100 million are at risk. This disease is considered a tropical neglected
18.0 ± 0.0, 11.0 ± 0.4, 20.0 ± 0.8, 20.0 ± 0.4, 10.0 ± 0.8 and one by WHO, but people affected by Chagas disease were already found
20.0 ± 0.9, respectively. These values show that the oleoresin col- in Spain, USA and others. Another important issue to be considered is
lected from different Copaifera species is a promising natural product, the efficacy of the treatment available in the market: they are effective
especially the one from C. reticulata, for the development of novel during the acute phase, although they are quite not efficient in the
pharmaceuticals aiming the treatment of leishmaniosis, a neglected chronic phase. Besides, it may cause several side effects [80].
disease, considering their low IC50 values. Also, C. reticulata have dis- Regarding Copaifera spp., scientific reports show the efficacy of this
played a promising activity against L. amazonensis axenic amastigotes natural product against Trypanosoma: C. martii oleoresin was effective
and intracellular amastigotes (IC50 of 15 and 20 μg/mL, respectively), in inhibiting different stages of T. cruzi growth, the ethologic agent of
the morphological and ultrastructural alterations were analyzed. The Chagas disease, with IC50 of 19 μg/mL for epimastigotes, 97.5 μg/mL
oleoresin promoted mitochondrial and plasma membrane damages. for trypomastigotes and less than 5 μg/mL for amastigotes (intracellular
Besides, it also caused a significant effect on the mitochondrial mem- form), apparently due to disruption and disorganization of the cell
brane potential and plasma membrane. membranes, which led to a reduction of the parasite dimensions. C.
Dos Santos et al. [78] tested C. martii’s oleoresin in vivo: topical and reticulata, C. cearensis, C. paupera, C. landorffii, C. multijuga, C. officinalis
oral formulations with this product were tested against lesions caused and C. lucens oleoresins also displayed a good activity against amasti-
by L. amazonensis in footpads of BALB/c mouse, and a reduction of the gotes (IC50 ≤ 10 μg/mL) [81].
lesion size was observed with oral treatment at a dose of 100 mg/kg/ In vitro and in vivo experiments against Trypanosoma evansi revealed
day, with neither changes in the histopathology of the tissue nor gen- the effect of the oleoresin from C. duckei against this parasite: in vitro
otoxic effect at a 2000 mg/kg dose. The samples promoted a similar tests showed a decrease in the parasites survival at 0.5, 1.0 and 2.0% of
response regarding to the lesion sizes in comparison to the positive oleoresin, and in vivo tests showed a dose-dependent trypanocidal ef-
control (Glucantime®) in the same dose. Considering that the oleoresin fect, displaying an extent in the longevity of the treated mice. This ef-
is a natural product and that it has a biological effect similar to one fect was due to the increasing of the membrane permeability and de-
reference drug in the market, it can be a future novel herbal medicine polarization of the mitochondrial membrane in T. evansi cells caused by
for leishmaniosis treatment after further drug development studies. the oleoresin. In vivo studies revealed that the biological effect is
Another important observation of this study was the confirmation of probably susceptible to the chemical composition [82]. Baldissera et al.
the morphological and ultrastructural changes in parasite cells treated [83] also assessed the oleoresin from C. duckei against T. evansi in vivo:
with IC50 of 14.0 μg/mL and IC90 of 70.0 μg/mL, which led to mi- this oleoresin at the dose 1.0 mL/kg increased the longevity of the
tochondrial swelling, plasma membrane permeability and depolariza- treated mice.
tion in the mitochondrial membrane. Therefore, the oleoresin of C. As an attempt to elucidate the mode of action of the trypanocidal
martii may contain compounds with pharmacological potential against compound 155 and the copaiba oil (C. martii), Kian et al. [84] reported
relevant pathologies, such as cutaneous leishmaniosis caused by L. that these samples were able to induce an anti-inflammatory effect by
amazonensis, since the medications available have high cost, are rela- increasing of IL-10 and protein growth factor beta (TGF-β) and reduc-
tively toxic and/or requires parenteral administration. tion of IL-12. They also found that the parasites death was not related to
The isolated compounds 185 and 221 showed high activity against nitric oxide or reactive oxygen species production, but probably they

12
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

are inducing a direct response on Trypanosoma. effect, which may induce the antitumoral response.
By taking that into account, it is possible to consider Copaifera spp.
oleoresin for development of novel food supplement or as new alter-
native source for compounds with trypanocidal activity. 5.5. Gastroprotective

5.4. Anticancer C. langsdorffii is the most studied species for this biological activity,
and one of the first studies used ethanol, indomethacin and hy-
Antitumor activity of C. multijuga was assessed by Gomes et al. [85]: pothermic restraint-stress models of induced gastric lesion in rats to test
the oleoresin, its hexanic and chloroformic fractions were tested against C. langsdorffii gastroprotection [90]. Oral administrations of this
Ehrlich tumor evolution: oral doses of 100, 150 and 200 mg/kg showed oleoresin at doses of 200 and 400 mg/Kg provided dose-dependent
antineoplasic effect against ascitic tumor and solid Ehrlich tumor after protection against gastric damage on the ethanol and restraint stress,
ten consecutive days of treatment. It promoted inhibition of solid tumor while on the indomethacin model, the dose of 400 mg/kg prevented the
on paws, reduced the production of several inflammatory mediators gastric ulceration. In all the cases there was a reduction of the lesion in
(total protein, PGE2, nitric oxide, and TNF) on ascitic fluid, as well as comparison with the cimetidine control at 100 mg/Kg.
blood and medulla cell count to values similar to control group. The C. The hydroalcoholic extract of C. langsdorffii leaves and the isolated
multijuga oleoresin at 150 mg/kg after 3 days of treatment showed a compounds 16, 12, 2, 155, 177 and 178, were also evaluated [8]: the
similar effect of vincristine (at 0.5 mg/kg) and after 4 days it was ap- extract (50, 250 and 500 mg/kg) displayed a reduction of the injured
proximately 30% better. Regarding the hexanic fraction, after 4 days of area caused by ethanol/HCl and indomethacin-induced ulcer model.
treatment, it induced a greater reduction in the tumor than the positive However, on the second model the doses did not reach the same
control at 150 and 200 mg/kg. The chloroformic fraction displayed a curative index of the cimetidine control (100 mg/kg). On the ethanol
significant antitumoral effect as well: after 4 days in all tested con- model, the curative index was higher in all cases, but the doses were
centrations (100, 150 and 200 mg/kg) the sample displayed a better elevated compared with the omeprazole control (30 mg/kg). The assays
effect than vincristine. with the isolated compounds (30 mg/kg) showed reduction of the ul-
Methanol extract from leaves of C. langsdorffi showed cytotoxicity cerative lesion, but only the terpenes had a good effect in comparison
against four tumor cell lines: B16 (murine skin), MCF-7, and HCT-8 with the cimetidine.
showing IC50 of 37.1 μg/mL, 12.9 μg/mL and 43.3 μg/mL respectively Galloylquinic acid derivative compounds (30 mg/kg) isolated from
[86]. C. langsdorffii leaves hydroalcoholic extract also displayed a the aqueous fraction of the leaf extract also promoted a great decreasing
protective effect against colon carcinogenesis according to Senedese in the lesion and showed a similar effect in the cure than pantoprazole
et al. [87]: wistar rats treated with oral doses of 20, 40 and 80 mg/kg at the same doses, especially the 4,5-dissubstituted galloylquinic acids
body weight of C. langsdorffii extract combined with 1,2-dimethylhy- [12].
drazine (DMH), a colon carcinogen, had significantly lower frequency Another studied species was C. malmei: its infusion leaves extract
of DNA damage (from 14.30% to 38.8%) in comparison with the po- showed prevention on the acute induced gastric lesions on the ethanol/
sitive control (DMH only). The effect was non-significant dose-depen- HCl model (400 mg/Kg) with similar action than carbenoxolone
dent. Furthermore, the groups treated with 40 and 80 mg/kg of leaves (100 mg/kg). Besides, it induced reduction of gastric secretion at
extract during and after DMH treatment displayed lower numbers of 200 mg/Kg, similar to cimetidine (50 mg/kg), and stimulation of gastric
genomic damage and aberrant crypt foci (ACF), in comparison with mucus production, which improved chronic ulcers healing process.
control. Regarding the isolated compounds, 200, 193 and 212 from C. Additionally, a reduction/prevention of epithelial and vascular damage
langsdorffii oleoresin were tested against Invasive Mammary Carcinoma was noticed at 100 and 400 mg/kg doses. On the other hand, when
(IMC) in mice, and 200 was the most potent compound, showing twice compared to known positive controls, the needed doses to induce these
effectiveness by measuring the increasing in life span at 41 mg/Kg/day effects were elevated [48].
for four 4 days in comparison with the positive control 5-fluorouracil
(30 mg/Kg/day, 4 days) [88]. Kaurenoic acid (155) at 78 μM produced
95% growth inhibition of CEM leukemic cells, and 45% of inhibition 5.6. Insecticide
MCF-7 (human breast) and HCT-8 (human colon) cancer cells [89]. This
diterpene also inhibited approximately 28% of tumoral cells line AGP01 Several Copaifera species revealed an interesting potential against
(gastric) and SF295 (human glioblastoma) viability after 72 h of ex- insects and an outstanding one is the action against vectors of important
position at 20 μmol/L [68]. On the other hand, 221 was cytotoxic on P- tropical diseases. Larvicidal effect against Aedes aegypti, the vector of
388 (DBA/2 mouse lymphoid neoplasm- IC50 of 2.5 μg/mL), A-549 dengue, Zika fever and chikungunya was induced by some Copaifera
(human lung carcinoma- IC50 of 5.0 μg/mL), HT-29 (colon carcinoma especies, such as C. multijuga: the ethanolic extracts of bark and leaves
Human- IC50 of 5.0 μg/mL) and MEL-28 (human melanoma- IC50 of of C. multijuga showed LC50 values of 81 and 166 mg/L, for the oleor-
10.0 μg/mL) [40]. Diterpene 184 showed cytotoxic activity to human esin it was 93 mg/L [91]. Apart from that, some isolated compounds
cervical adenocarcinoma (HeLa), human glioblastoma (MO59 J) and and other Copaifera species are even more detachable. One example is
human breast adenocarcinoma (MCF-7), with IC50 of 44.03 μg/mL, the compound 185 isolated from the C. multijuga oleoresin that pre-
68.3 μg/mL, and 150.3 μg/mL, respectively [57]. sented a LC50 value of 0.7 mg/L [91], while C. langsdorffii oleoresin was
By analyzing these reports, it can be observed that the Copaifera even more promising with an LC50 of 0.041 mg/L [92].
oleoresin in several in vivo experiments demonstrated a better effect Action against larvae of the main vector of malaria on Brazil
than positive controls. Some isolated compounds, such as kolavenol, Anopheles darlingi was reported for C. multijuga ethanolic extracts of the
also displayed a significant activity in comparison with positive con- bark and leaves with LC50 of 3 and 13 mg/L respectively. The terpenoid
trols. Concerning the in vitro experiments, considering potent cytotoxic 185 showed similar action at 0.9 mg/L [91].
compounds with IC50 lower than 10 μM, methyl copalate is a promising The insecticidal activity against pests was also assessed: extracts
one. On the other hand, kaurenoic and copalic acids did not show a from fruit peels and leaves of C. langsdorffii tested in concentrations
potent activity. Hence, there are antitumor/cytotoxic compounds in from 300 mg/L have increased mortality of first instar larvae, and ad-
Copaifera oleoresins, although some major isolated diterpenes do not verse effects in other stages like reduction on the caterpillar’s weight,
present a significant activity. Probably, the compounds responsible for reduction of growth of second instar larvae, decreasing of oviposition
this citotoxic effect may not be the known diterpenes or they, along period and in number and viability of eggs [93].
with the other ones in the samples, may act in a synergic/additive

13
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

5.7. Other activities 6. Quality control

There are several studies about a miscellaneous of biological and Since the biological activities described for Copaifera oleoresin and
pharmacological effects of Copaifera oleoresins, its isolated compounds other parts extracts are directly related to their chemical composition,
and/or derivatives. One approach is regarding to the reproductive its quality control is mandatory. In the development of phytotherapic
system: a nanoformulation containing C. langsdorffii oleoresin (PVP- medicines, it is necessary to standardize the product, qualitatively and
K30:clay: oleoresin - [Link]) was tested in vitro (50 μg/mL), for the quantitatively, to be approved by Surveilance Agencies due to the
treatment of endometriosis, causing changes in the morphology and correlation between efficacy and safety and chemical composition.
promoting apoptosis and necrosis process on ectopic endometrium Besides, the adulteration of the oleoresin is frequently performed by
stromal cells. This effect is related to the increased lactate dehy- adding other types of oils to increase the volume of the material and
drogenase into the extracellular medium [94]. consequently earn more money from the sales of the oleoresin.
A relaxant effect on the smooth muscle on rat uterus was observed Therefore, research in this field has been increasing in the last decades:
when the tissue was exposed to 160 μM of 155 [95]: a contractile re- Cascon and Gilbert [17] aiming to identify the main compounds in C.
sponse of the muscles was stimulated by oxytocin and acetylcholine on guyanensis Desf., C. duckei Dwyer and C. multijuga Hayne, analyzed
the uterus in vitro and more than 80% of the response was inhibited in samples of the oleoresins by gas chromatography coupled to mass
two biological models. The biological action is attributed to calcium spectrometry. Gas chromatography is a useful technique since it allows
blockade and opening of ATP-potassium channels. the analysis of compounds from several classes of natural products,
C. langsdorffii oleoresin also showed a protective action against in- requiring volatility from the analytes. It can be achieved by derivati-
duced damage of intestinal tissue by ischemia/reperfusion process (I/ zation reactions, when the compound(s) do(es)not have this physical
R): oral treatment with 200 and 400 mg/Kg of the oleoresin attenuated property. This technique, when coupled to mass spectrometry has even
the I/R antioxidant and anti-lipid peroxidation mechanism, diminishing more value, since mass spectrometry gives information regarding the
myeloperoxidase (MPO), malondialdehyde (MDA) and catalase (CAT) mass and fragmentation of each analyzed compound, and it can still be
activities, nitric oxide (NO) level, and prevented glutathione (GSH) compared to libraries, providing even more information about the
diminution [96]. The effects on I/R of C. langsdorffii oleoresin were also identity of the compounds.
studied, but with skin flaps model on rats dorsums [61]: the oleoresin at Tappin et al. [99] also took advantage of gas chromatography
200 mg/kg and 400 mg/kg was administered by gavage 24, 12, and 2 h technique, by developing a suitable method for quality control of Co-
prior to the beginning of the procedure. The sample doses were ad- paifera oleoresin in gas chromatography with a flame-ionization de-
ministered daily during a week. Decreases in the tissue concentration of tector, since this detector gives a similar response to compounds from
tiobarbituric acid-reactive substances (TBARS) were observed, in- the same class. This method allows the analysis of methylated Copaifera
dicating that the oleoresin protects against lipid peroxidation, ex- oleoresins. Trans-(-)-caryophyllene (12) or methyl copalate (221) were
hibiting an antioxidant action. Significant increase in plasmatic and used as external standards to perform quantification of the sesqui-
tissue GHS levels were observed, as well as a significant decrease in terpenes and acid diterpenes in this product, respectively, allowing the
MPO concentrations in the tissues, suggesting an additional anti-in- analysis of several commercial Copaifera oleoresins. Terpene 12 is a
flammatory activity of the oleoresin. suitable external standard to perform quantification of total sesqui-
On the skin, an initial clinical assay demonstrated the power of a terpenes, especially when the analyzed sample has sesquiterpenes as
preparation based on C. langsdorffii oleoresin (1%) on attenuating acne major compounds, and in the same way, 221 to quantify acid di-
after 21 days of treatment [97]. Another initial clinical trial demon- terpenes. Methods such as this one, are important tools in the quality
strated that the oleoresin of C. langsdorffii also displayed effect against control of Copaifera oleoresin since it can be useful in the identification
chronic psoriasis, for which the oral intake (1 mL/day as dose) was of the Copaifera species and in the quantification of sesquiterpenes and
more effective than the topical administration in decreasing erythema, diterpenes, which comprises most of the identified classes of com-
skin thickness, and scaliness after six weeks of treatment [25]. pounds in Copaifera species. Besides, 12 and 221 are two important
Considering the potential of treating urinary disorders, C. langs- Copaifera markers and the presence of these compounds probably in-
dorffii aerial parts extract was tested to assess the effect on urolithiasis dicates that the oleoresin came from Copaifera spp. Therefore, it also
induced by CaOx pellet insertion in mice bladder: it was found that allows the identification of mixture of oleoresins from different species,
treatment with this extract (20 mg/Kg b.w) did not modify renal since their chemical composition (qualitatively and/or quantitatively)
function of the animals and provoked an increasing in the magnesium usually differs from one species to another. This is pertinent from
levels, which possibly led to decreasing in the formation of calcium quality control point of view due to the extent use of this product in
oxalate crystals [11]. Besides, it was observed a reduction in the ex- several fields, such as cosmetic, pharmaceutical, foods and others.
cretion of uric acid, which means that it may be acting on diminishing Biavatti et al. [100] also aiming to improve quality control of Copaifera
or blocking the formation of uric acid stones. oleoresins analyzed several samples from different places of Brazil by
Also, an in vivo model was used aiming to assess the potential of gas chromatography with a flame ionization detector and thin layer
leaves extract of C. langsdorffii in the treatment of nephrolithiasis. The chromatography. Some diterpenes found in Copaifera species were used
extract administered orally (1.6 mg/mL) prevented stone formation, as standards, such as 193, 184, 155 and 26. The authors also compared
compared with the ethylene glycol control (1% v/v), and diluted the their results with analysis by using an alternative acidity index method
crystals in vitro [98]. described by Vasconcelos and Godinho [101] and found that the last
In the assessment of these other activities, most of the authors did one did not provide reliable results for the analysis of Copaifera oleor-
not compare the effect of the samples with positive controls, which esins. Barbosa et al. [102] analyzed numerous samples of Copaifera
makes it difficult to evaluate if the activity is significant indeed. Despite oleoresins bought in the market and compared both the refractive index
of that, they have compared it with negative controls. Despite of lacking and the analysis by thin layer chromatography with a standard oleor-
positive controls, these reports indicates some biological response and, esin and found that from the twelve bought samples, three showed
in many cases, in low doses/concentrations. Therefore, it is worthy to retention index (RI) and chromatographic profile similar to the stan-
further study Copaifera oleoresin and its isolated compounds for several dard, six showed RI and chromatographic profile from soybean oil and
activities to find out in which cases the samples are the most promising three a chromatographic profile similar to the mixture of Copaifera
ones. oleoresin and soybean oil: two in the proportion of 3:1 and one in 1:3.
Therefore, the adulteration of this product frequently happens and
quality control is important to assure the identity and purity of the

14
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

oleoresin. quantification of 12 and 16, two of the main compounds found in

Because of the variations regarding the amount and identity of the several Copaifera species. They applied Box-Behnken design to select
main sesquiterpenes and diterpenes in Copaifera oleoresins of different the chromatographic parameters that provides the better results like
species and the complexity of the chemical composition, usually ana- peak symmetry and resolution of the peaks. After developing the
lysis by Thin Layer Chromatography and other simple methods are used method, it was validated according to international parameters
for an initial screening in quality control. When it is necessary to dis- showing that it is adequate to perform analysis and quantification of
tinguish the Copaifera species and consequently which diterpenes and these compounds.
sesquiterpenes are present and its respectively amounts, methods that Silva et al. [105] describes a validated method for quantification of
are capable of assuring these data should be employed: Sousa et al. [10] six diterpenes 164, 194, 237, 184, 185 and 186 in Copaifera oleoresin
reported a validated method in gas chromatography with a flame io- of several species, which is possible due to the similar chemical com-
nization detector able to quantify the volatile terpenes 12, 9, and 16 in position of such plant species. They were isolated from C. duckei, C.
different Copaifera oleoresins. The developed method presented se- reticulata and C. multijuga and comprise the major ones in such oleor-
lectivity, linearity from 0.10 to 18.74 mM, precision, accuracy, low esins. The method is a RP-HPLC one, using a PDA detector at 201 nm
limit of detection, satisfactory recovery of the compounds and standard and an isocratic mixture (8:2) of acetonitrile and acid water was se-
deviation lower than 10%. Therefore, it is suitable for analyzing Co- lected as mobile phase. Therefore, this is a simple method, which can be
paifera samples by quantifying these three sesquiterpenes. used by Universities and Companies aiming at analyzing Copaifera
Barbosa et al. [103] with the goal of studying Copaifera species oleoresins and performing its quality control.
chemical variability, analyzed 22 oleoresin samples of C. multijuga, after Dias et al. [109] and Lucca et al. [110] developed formulations with
esterification, by gas chromatography with a flame ionization detector copaiba oleoresin extracted from C. multijuga. The first ones developed
and GC coupled to a mass spectrometer. They found 35 compounds, and nanoemulsions aiming to treat inflamed skin. After extraction by HS-
after statistical analysis, the samples were grouped into three similar SPME, the stability of 12 in this formulation was assessed by a devel-
groups. 12 and 2 were the main sesquiterpenes detected in all samples, oped and validated gas chromatography method with a flame ioniza-
in different amounts, and the ones that are important to classification of tion detector. It was also used in the quantification of this compound in
the oleoresins. It was also observed that different chromatographic this formulation. The second ones studied the absorption from the skin
profiles of the oleoresins from the same species of Copaifera may occur, of copaiba oleoresin released by a developed nanoemulstion, and they
even if they were cultivated under similar environment. Wong et al. developed and validate a HS-GC–MS method to quantify 12 both in the
[104] aiming to perform qualitative analysis of the oleoresin, used two nanoemulsion and in the crude oleoresin.
dimensional gas chromatography coupled to a Time-of-flight mass Although several authors have developed and validated methods to
spectrometer combining a more polar phase with a lower one in the perform quality control of the oleoresins, the challenge still remains,
second phase to analyze sesquiterpenes and acid diterpenes. because there is chemical variation inter and also intra species, espe-
Regarding the non-volatile compounds, a method developed by cially regarding the amount of the sesquiterpenes and diterpenes found
Motta et al. [12] using HPLC-UV can be applied in quality control of C. in the different oleoresins. On the other hand, some chemical markers
langsdorffii leaves, since it allows the analyses and quantification of nine such as 12 and 184 are usually present in most Copaifera species, and
galloylquinic acid derivatives and the flavonoids 177 and 178, which therefore, can be considered for evaluation of adulteration of the
are the major compounds found on the leaves. To analyze them, the oleoresin with soybean, for example. Another important perspective is
author chose as stationary phase a Synergi Polar-RP column (Phe- from the biological activities displayed by the oleoresins and/or their
nomenex), an ether-linked phenyl column: this property is responsible isolated compounds: by quality control of the samples, several biolo-
for increasing the aromaticity. Hence, it allows π–π interactions with gical effects of the oleoresins can be assured for specific samples, be-
insaturated compounds, allowing more interaction with the column and cause biological activities are usually correlated with the chemical
better separation of the chromatographic peaks of polar and aromatic profile of the samples.
compounds.
Da Silva et al. [105] developed and validated an UPLC-MS/MS 7. Toxicology
method to identify and quantify nine acid diterpenes in the oleoresin of
several Copaifera species, both the ones collected in different Brazilian Regarding Copaifera oleoresins, several reports described a few as-
geographic places and commercially obtained. This method is suitable pects of its safety: the genus Copaifera L. is on the list of medicinal
and reliable for quality control analyses of the oleoresin because acid plants contraindicated in pregnancy or lactation, according to the State
diterpenes are the major compounds in this product and the method Program for Medicinal Plants of the State Health Secretariat of Rio de
was validated, which proves its reliability. Janeiro [111]. To evaluate if this toxic effect is accurate, we have
Also, an HPLC method to quantify five phenolic compounds 163, compared it with several publications: in vivo studies of the ethanolic
177 and 171 in hydroethanolic extracts of C. langsdorffi leaves was extract of the bark of C. luetzelburgii indicated an increasing on the
developed by Sousa et al. [106]. They used as stationary phase two relative weight of the uterus of rats with oral doses of 125, 250 and
monolithic columns linked in series (100 x 4.6 mm - C18) and elution by 500 mg/kg [111]. Also, treatment with the extract during the preg-
gradient. Since phenolic compounds have chromophores, UV detection nancy did not induce toxicity in the progenitors or progeny; however, it
was performed at 275 nm. Moreover, the method was validated to as- caused shortening of pregnancy at 125 mg/kg. In the in vitro study,
sure its reliability. Souza et al. [107] aiming to quantify 184, one Copaifera oleoresin inhibited isometric cells after induction of con-
chemical marker of Copaifera genus, also developed an efficient RP- tractions in pregnant and non-pregnant rats. The influence of C. multi-
HPLC method and validated it according to ANVISA, 2003. One of the juga oleoresin on reproductive parameters was studied as well, de-
advantages of this method, when compared to some gas chromato- monstrating that oral administration of the oleoresin at doses of 200,
graphy methods, is that it is not necessary to perform derivatization 500 and 2500 mg/kg on male Wistar rats, didn’t affect fertility rates,
reactions before the analysis. Besides, the authors observed that the absolute or relative mass of accessory sexual organs, histology or de-
chromatographic profile and the amount of 184 are very different velopment of the offspring [112].
among different analyzed samples, which suggests that they probably Besides, the effect of a vaginal cream containing C. duckei oleoresin
have been adulterated and/or oleoresins from distinct Copaifera species on reproductive performance of female Wistar rats was assessed: the
have been mixed. oleoresin was tested at 28.6 mg/kg in the vaginal cream and the
A novel RP-HPLC method was developed by Borges et al. [108] treatment lasted from one month before until 20 days of pregnancy. The
using a cyano column and acetonitrile phosphate buffer to perform the authors neither found signs of toxicity or malformations to the fetus nor

15
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

to the mother at this dose [113]. samples collected after 24 h and 36–56% in samples collected after 48 h
Nevertheless, other studies about the embryotoxic and the toxicity of the genotoxic solution administration) in comparison with animals
to pregnant female rats of C. reticulata oleoresin administered orally receiving doxorubicin alone [116]. Also, 100 mg/kg of C. langsdorffii
revealed toxicity at high doses: at 1000 and 1250 mg/kg/day, the body fruit powder showed a 38% reduction of micronuclei in mice ery-
weight of the offspring was reduced, and the kidneys and ureters of the throcyte [24].
newborns had minor deformities. Altough, it was not clearly related to Similarly, C. malmei leaves extract was not cytotoxic to normal cells
the dose. At the highest tested doses some bone variations had in- such as CHO-k1 cells in vitro, with an IC50 value of 4200 mg/mL after
creased, and development of the fetuses decreased. On the other hand, 24 h and 72 h of exposure. In vivo, it did not present oral acute toxicity
no teratogenic effect was observed. This reports have estimated the in mice at a 5000 mg/kg dose [48].
safety dose for pregnant woman at 5 mg/kg/bw/day and a re- Therefore, based on the reported toxicological data, the use of
commended dose of 0.0125 mg/kg/bw/day [43]. Copaifera oleoresin does not seem to be genotoxic, mutagenic or cyto-
In vitro studies on the sea urchin egg development were performed toxic. However, at higher doses it was reported the LD50 of approxi-
with the methanol extract from leaves of C. langsdorffi, showing in- mately 4 g/kg for C. reticulata and C. multijuga [78], the oleoresin of
hibition of first cleavage in sea urchin egg by 80% at 100 μg/mL [86]. Copaifera spp may display considerably toxic effects. On the other hand,
Also the isolated compound 155 showed inhibition of first cleavage of the administration of the oleoresin during pregnancy is not re-
sea urching egg with an IC50 of 84.2 μM, and progressively induced commended since it is responsible for decreasing the fetus body weight
embryo destruction at 44.7 and 10 μM for blastulae and larvae stages, and cause malformation on kidneys and ureters in mice at 1000 mg/kg/
respectively [89]. day [43]. Besides, extracts of other parts of Copaifera species did not
Furthermore, in vivo genotoxic assays of a commercial Copaifera reveal toxic properties, and some of them even showed potential cito-
oleoresin in which 12 was the major compound, were performed and protection against DNA harmful agents [24,116].
[114] at 500, 1000 and 2000 mg/kg administered orally, it neither
increased DNA damage nor micronucleated polychromatic ery- 8. Conclusion
throcytes, which means that this oleoresin did not show significant
mutagenic, genotoxic or cytotoxic effect on bone marrow cells. The Some Copaifera species are among the most studied ones in the
tested doses of these and several other studies were probably based on world due to their economic and ecological importance. Throughout
previous report [42], who showed a LD50 of 3.9 and 4.3 g/kg for C. the years, it was attributed important biological activities to Copaifera
reticulata and C. multijuga, respectively. Also, safety of C. martii oleor- spp against several pathologies, which further reinforces the greatness
esin on mammalian cells was reported [78]: treated animals with of this genus.
2000 mg/kg of the oleoresin didn’t demonstrate any mutagenic effects, Despite the many reported publications for Copaifera genus, there
nor behavior, clinical or tissue alterations. are still many important issues to be addressed by other fields. For
Regarding the antiproliferative effect of C. duckei oleoresin, the oral example, the most studied species of Copaifera are C. langsdorffii, C.
gavage of 600 mg/kg/bw/day for seven days in rats, showed inhibition multijuga, C. officinalis and C. reticulata. On the other hand, we have not
of the proliferation of hepatocytes after 24 h of partial hepatectomy found any publication on the following species: C. aromatica Dwyer, C.
and, apparently affecting the energy metabolism of the hepatocytes baumiana Harms, C. bracteata Benth., C. brasiliensis Dwyer, C. bulbo-
mitochondria [115]. Besides, rats were also used to evaluate the mu- tricha Rizzini & Heringer, C. camibar Poveda, Zamora & Sanchez, C.
tagenic and cytotoxic effect on peripheral blood reticulocytes and bone canime Harms, C. depilis Dwyer, C. elliptica Mart., C. galedupa (Prain)
marrow cells of C. duckei oleoresin at 10, 25 and 50% of the LD50 after Oken, C. grandifolia (Benth.) Malme, C. gynohirsuta Dwyer, C. jac-
0, 24, 48 and 72 h [37]. No significant divergence between the mean quiniana G. Don, C. jacquinii Desf., C. jussieui Hayne, C. laevis Dwyer, C.
number of micronucleated reticulocytes or chromosomal aberrations of magnifolia Dwyer, C. majorina Dwyer, C. marginata Benth, C. mildbraedii
the oleoresin and the negative control was found. However, at 25 and Harms, C. nana Rizzini, C. rondonii Hoehne, C. sabulicola [Link] &
50% of the LD50, the oleoresin reduced significantly the mitotic index. L.P. Queiroz., C. utilissima Saldanha and C. rondonii Hoehne. Therefore,
Therefore, it was not mutagenic, although at higher doses, it seemed to the number of species that has not been studied shows the huge gap to
be cytotoxic. be filled by researchers of different fields to fully determine the po-
Kaurenoic acid (155), isolated from C. langsdorffii oleoresin, caused tential of Copaifera genus for pharmaceutical, cosmetics and food in-
toxic effects on chinese hamster lung fibroblast (V79), a normal cell dustries, among others.
model at concentrations of 30 and 60 μg/mL. This diterpene caused Overall, it can be concluded that there are many compounds and
significant increase in cell damage index and frequency, indicating that oleoresins reported here with great potential for further studies, aiming
would induces genotoxicity. However, 155 did not show cytotoxicity to to develop new medicines either with standardized extracts or pure
normal cell lines murine fibroblast 3T3-L1 (at 5 mg/mL), human lym- compounds: by taking into account the reports of cytotoxicity of the
phocytes (500 μM), or J774 murine macrophage (IC50 > 100 μM). oleoresins, C. multijuga demonstrated to have a great potential, since it
Moreover, no significant hemolytic activity (IC50 86.7 mM) was ob- displays a better effect than vincristine, a potent known anticancer
served [68]. agent. Considering the pure compounds, 200 and 221 show low values
Toxicology of other parts of the plants were also studied: the me- of IC50 (below 10 μg/mL), which indicates their promising activity.
thanolic extract from the leaves of C. langsdorffii displayed an EC50 of Regarding the antiparasitic potential of Copaifera spp., considering as
544.5 μg/mL on mouse erythrocytes and caused hemolytic effect on good effect samples with IC50 values below 10 μg/mL, trypanocidal
these cells [86]. Besides, another study revealed that C. langsdorffii leaf oleoresins are the following ones: C. martii, C. reticulata, C. cearensis, C.
extract at 80 mg/kg does not display genotoxicity tested by comet assay paupera, C. langsdorffii, C. multijuga, C. officinalis and C. lucens.
[87]. The results did not show any significant differences between Regarding leishmanicidal effect, besides C. reticulata, C. multijuga and C.
treated animals and the negative control group (receiving EDTA only, martii oleoresins, the isolated compounds 12, 186, 221, 198 and 155
p > 0.05). have IC50 values lower than 6 μg/mL. It was also reported a good effect
The hydroalcoholic extract of C. langsdorffiii leaves decreased the against Plasmodium falciparum promoted by C. religiosa oleoresin.
number of micronucleus induced by the genotoxic agent doxorubicin on In relation to wound healing effect, some studies showed the po-
the peripheral blood micronucleus assay. Administration of 10, 20, 40 tential of C. langsdorffii oleoresin in reducing the lesion size efficiently.
and 80 mg/kg of the extract on Swiss mouse for 20 days pointed that on To our knowledge, isolated compounds with wound healing effect have
the treatments combining the extract and the genotoxic agent, the not yet been evaluated. Likewise, their anti-inflammatory and anti-
number of micronucleus had significantly decreased (59–77% in nociceptive activities have not been assessed as well, although C.

16
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

multijuga, C. officinalis, C. langsdorffii, C. cearensis display a significant 475–481, [Link]

anti-inflammatory effect in comparison with positive controls. [12] E.V.S. Motta, M. Lemos, J.C. Costa, V.C. Banderó-Filho, A. Sasse, H. Sheridan,
J.K. Bastos, Galloylquinic acid derivatives from Copaifera langsdorffii leaves dis-
Therefore, it can be concluded that Copaifera spp. and several of their play gastroprotective activity, Chem. Biol. Interact. 261 (2017) 145–155, https://
isolated compounds showed promising activities against different [Link]/10.1016/[Link].2016.11.028.
cancer cells, parasites and body reactions, such as inflammation. [13] K.C.O. de Albuquerque, A. do S.S. da Veiga, J.V. da S. e Silva, H.P.C. Brigido,
E.P. dos R. Ferreira, E.V.S. Costa, A.M.D.R. Marinho, S. Percário, M.F. Dolabela,
Despite of that, in most cases, only the initial studies have been per- Brazilian amazon traditional medicine and the treatment of difficult to heal
formed and to really prove the efficacy and potency of these samples, Leishmaniasis wounds with Copaifera, Evidence-Based Complement, Altern. Med.
aiming at novel drug developments, further studies should be per- 2017 (2017) 1–9, [Link]
[14] P.L. Tobouti, T.C. de Andrade Martins, T.J. Pereira, M.C.M. Mussi, Antimicrobial
formed. activity of copaiba oil: a review and a call for further research, Biomed.
Moreover, modes of action should be fully determined, pharmaco- Pharmacother. 94 (2017) 93–99, [Link]
kinetic and pharmacodynamic studies should be undertaken, as well as [15] A.L. Diefenbach, F.W.M.G. Muniz, H.J.R. Oballe, C.K. Rösing, Antimicrobial ac-
tivity of copaiba oil (Copaifera spp.) on oral pathogens: systematic review,
clinical trials with products produced by pharmaceutical companies.
Phyther. Res. (2017) 586–596, [Link]
From the phytotherapic development point of view, despite of the [16] Missouri Botanical Garden, Copaifera L., [Link]. (2017). [Link]
development of several analytical methods for analyses of Copaifera [Link]/[Link]?nameid=40013764&tab=subordinatetaxa
oleoresin, a difficult challenge is to standardize the chemical compo- (Accessed 30 March 2017).
[17] V. Cascon, B. Gilbert, Characterization of the chemical composition of oleoresins
sition, since although most of the chemical compounds in the samples of Copaifera guianensis Desf., Copaifera duckei Dwyer and Copaifera multijuga
are sesquiterpenes and diterpenes, their presence and amounts sig- Hayne, Phytochemistry 55 (2000) 773–778, [Link]
nificantly differ from one species to another, and even among in- 9422(00)00284-3.
[18] V.F. Veiga, L. Zunino, J.B. Calixto, M.L. Patitucci, Â.C. Pinto, Phytochemical and
dividuals of the same species, which affects the biological activities. antioedematogenic studies of commercial copaiba oils available in Brazil, Phyther.
Regarding the safety use of Copaifera spp., we conclude that in Res. 15 (2001) 476–480, [Link]
therapeutic doses, most oleoresins do not display genotoxic, mutagenic [19] A.C. Pinto, W.F. Braga, C.M. Rezende, F.M.S.S. Garrido, V.F. Veiga Jr, L. Bergter,
M.L. Patitucci, O.A.C. Antunes, Separation of acid diterpenes of Copaifera cearensis
or cytotoxic effects on normal cells. However, to guarantee the safe use Huber ex Ducke by flash chromatography using potassium hydroxide impregnated
of these oleoresins, other studies must be performed. With the available silica gel, J. Braz. Chem. Soc. 11 (2000) 355–360, [Link]
data in the literature, there are not sufficient data regarding efficacy, S0103-50532000000400005.
[20] A.T. Oliveira-Filho, J.A. Ratter, A study of the origin of central Brazilian forests by
safety and quality, to allow the registration of novel pure compound or the analysis of plant species distribution patterns, Edinb. J. Bot. 52 (1995)
phytopharmaceutical medicine. However, there are promising pre- 141–194, [Link]
liminary results on several biological activities, in addition to tox- [21] M. Calvin, Hydrocarbons from plants: an analytical methods and observations,
Naturwissenschaften 67 (1980) 525–533.
icological reports and analytical methods, which may lead to the de-
[22] E.K. Nemethy, E.S. Lipinsky, Biochemicals as an energy resource, CRC Crit. Rev.
velopment of new hits in the pharmaceutical industry. Plant Sci. 2 (1984) 117–129, [Link]
[23] E. Stashenko, H. Wiame, S. Dassy, J.R. Martinez, T. Shibamoto, Catalytic trans-
Conflict of interest formation of copaiba (Copaifera officinalis) oil over zeolite ZSM‐5, J. High Resol.
Chromatogr. 18 (1995) 54–58, [Link]
[24] Â.G. Batista, A.S. Ferrari, D.C. da Cunha, J.K. da Silva, C.B.B. Cazarin, L.C. Correa,
The authors state no conflict of interest. M.A. Prado, L.B. De Carvalho-Silva, E.A. Esteves, M.R. Maróstica Júnior,
Polyphenols, antioxidants, and antimutagenic effects of Copaifera langsdorffii fruit,
Food Chem. 197 (2016) 1153–1159, [Link]
Acknowledgement 11.093.
[25] F. Gelmini, G. Beretta, C. Anselmi, M. Centini, P. Magni, M. Ruscica, A. Cavalchini,
Authors are thankful to São Paulo Research Foundation (FAPESP) R. Maffei Facino, GC-MS profiling of the phytochemical constituents of the
oleoresin from Copaifera langsdorffii Desf. and a preliminaryin vivo evaluation of its
for financial support, Grant 2011/13630-7. antipsoriatic effect, Int. J. Pharm. 440 (2013) 170–178, [Link]
[Link].2012.08.021.
References [26] A. Santos, T. Ueda-Nakamura, B. Dias Filho, V. Veiga Jr, A.C. Pinto,
C.V. Nakamura, Effect of Brazilian copaiba oils on Leishmania amazonensis, J.
Ethnopharmacol. 120 (2008) 204–208, [Link]
[1] V.F. Veiga Jr, A.C. Pinto, O gênero Copaifera L, Quim. Nova 25 (2002) 273–286, 007.
[Link] [27] N.V. Gramosa, E.R. Silveira, Volatile constituents of Copaifera langsdorffii from the
[2] J. Léonard, Notulae systematicae IV (Caesalpiniaceae-Amherstieae africanae Brazilian Northeast, J. Essent. Oil Res. 17 (2005) 130–132, [Link]
americanaeque), Bull. Du Jard. Bot. l’Etat Brux. 19 (1949) 383–407. 1080/10412905.2005.9698853.
[3] J.D. Dwyer, Further studies on the new world species of Copaifera, Bull. Torrey [28] J. de S. Lima Neto, N.V. Gramosa, E.R. Silveira, Constituintes químicos dos frutos
Bot. Club. 81 (1954) 179–187. de Copaifera langsdorffii Desf, Quim. Nova 31 (2008) 1078–1080, [Link]
[4] The plant list, Copaifera. (2013). [Link] 10.1590/S0100-40422008000500025.
copaifera (Accessed 30 March 2017). [29] T. Stupp, R.A. de Freitas, M.R. Sierakowski, F.C. Deschamps, A. Wisniewski,
[5] F.A. Pieri, M.C. Mussi, M.A.S. Moreira, Óleo de copaíba (Copaifera sp.): histórico, M.W. Biavatti, Characterization and potential uses of Copaifera langsdorffii seeds
extração, aplicações industriais e propriedades medicinais, Rev. Bras. Plantas Med. and seed oil, Bioresour. Technol. 99 (2008) 2659–2663, [Link]
11 (2009) 465–472, [Link] [Link].2007.04.051.
[6] R.C.V. Martins-da-Silva, J.F. Pereira, H.C. de Lima, O gênero Copaifera [30] M.E. do Nascimento, M. das, G.B. Zoghbi, J.E. Brasil Pereira Pinto, S.K. Vilela
(Leguminosae – Caesalpinioideae) na Amazônia brasileira, Rodriguésia 59 (2008) Bertolucci, Chemical variability of the volatiles of Copaifera langsdorffii growing
455–476. wild in the Southeastern part of Brazil, Biochem. Syst. Ecol. 43 (2012) 1–6,
[7] L.M. Leandro, F. de Sousa Vargas, P.C.S. Barbosa, J.K.O. Neves, J.A. da Silva, [Link]
V.F. da Veiga-Junior, Chemistry and biological activities of terpenoids from [31] J.F. Carmo, I. Miranda, T. Quilhó, V.B. Sousa, S. Cardoso, A.M. Carvalho,
Copaiba (Copaifera spp.) oleoresins, Molecules 17 (2012) 3866–3889, [Link] F.H.D.J. Carmo, J.V.F.F. Latorraca, H. Pereira, Copaifera langsdorffii bark as a
org/10.3390/molecules17043866. source of chemicals: structural and chemical characterization, J. Wood Chem.
[8] M. Lemos, J.R. Santin, C.S. Mizuno, T. Boeing, J.P.B. De Sousa, D. Nanayakkara, Technol. 36 (2016) 305–317, [Link]
J.K. Bastos, S.F. De Andrade, Copaifera langsdorffii: evaluation of potential gas- [32] L.F.R. de Almeida, R. de O. Portella, J. Bufalo, M.O.M. Marques, R. Facanali,
troprotective of extract and isolated compounds obtained from leaves, Braz. J. F. Frei, Non-Oxygenated sesquiterpenes in the essential oil of Copaifera
Pharmacogn. 25 (2015) 238–245, [Link] langsdorffiiDesf. increase during the day in the dry season, PLoS One 11 (2016)
[9] V.F. Veiga Jr., M.L. Patitucci, A.C. Pinto, Controle de autenticidade de óleos de e0149332, , [Link]
copaíba comerciais por cromatografia gasosa de alta resolução, Quim. Nova 20 [33] M.S. Nogueira, R.A. Furtado, J.K. Bastos, Flavonoids and methoxy-galloylquinic
(1997) 612–615, [Link] acid derivatives from the leaf extract of Copaifera langsdorffii Desf, J. Agric. Food
[10] J.P.B. Sousa, A.P.S.S. Brancalion, A.B. Souza, I.C.C.C. Turatti, S.R. Ambrósio, Chem. 63 (2015) 6939–6945, [Link]
N.A.J.C.J.C. Furtado, N.P. Lopes, J.K. Bastos, Validation of a gas chromatographic [34] G.B. Zoghbi, O. a Lameira, E.C.P. Oliveira, G.B. Zoghbi, O. a Lameira,
method to quantify sesquiterpenes in copaiba oils, J. Pharm. Biomed. Anal. 54 E.C.P. Oliveira, G.B. Zoghbi, M. das, G.B. Zoghbi, O. a Lameira, E.C.P. Oliveira,
(2011) 653–659, [Link] Seasonal variation of oleoresin and volatiles from Copaifera martii Hayne growing
[11] A.P.S. Brancalion, R.B. Oliveira, J.P.B. Sousa, M. Groppo, A.A. Berretta, wild in the State of Pará, Brazil, J. Essent. Oil Res. 19 (2007) 504–506, [Link]
M.E. Barros, M.A. Boim, J.K. Bastos, Effect of hydroalcoholic extract from org/10.1080/10412905.2007.9699316.
Copaifera langsdorffii leaves on urolithiasis induced in rats, Urol. Res. 40 (2012) [35] W.F. Braga, C.M. Rezende, O.A.C. Antunes, A.C. Pinto, Terpenoids from Copaiba

17
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

cearensis, Phytochemistry 49 (1998) 263–264, [Link] Júnior, C. Vataru Nakamura, Antileishmanial activity of diterpene acids in copaiba
9422(97)00899-6. oil, Mem. Inst. Oswaldo Cruz 108 (2013) 59–64, [Link]
[36] V.F.F. Veiga, E.C.C. Rosas, M.V.V. Carvalho, M.G.M.O. Henriques, A.C. Pinto, 02762013000100010.
Chemical composition and anti-inflammatory activity of copaiba oils from [57] F. Abrão, L.D. de Araújo Costa, J.M. Alves, J.M. Senedese, P.T. de Castro,
Copaifera cearensis Huber ex Ducke, Copaifera reticulata Ducke and Copaifera S.R. Ambrósio, R.C.S. Veneziani, J.K. Bastos, D.C. Tavares, C.H.G. Martins,
multijuga Hayne—a comparative study, J. Ethnopharmacol. 112 (2007) 248–254, Copaifera langsdorffii oleoresin and its isolated compounds: antibacterial effect and
[Link] antiproliferative activity in cancer cell lines, BMC Complement. Altern. Med. 15
[37] E.L. Maistro, J.C.T. Carvalho, V. Cascon, M.A.C. Kaplan, In vivo evaluation of the (2015) 443, [Link]
mutagenic potential and phytochemical characterization of oleoresin from [58] C. Arruda, D. de S. Eugênio, M.R. Moreira, G.V. Símaro, J.K. Bastos,
Copaifera duckei Dwyer, Genet. Mol. Biol. 28 (2005) 833–838, [Link] C.H.G. Martins, M.L.A. Silva, R.C.S. Veneziani, P.B. Vieira, S.R. Ambrósio,
1590/S1415-47572005000500028. Biotransformation of (-)-cubebin by Aspergillus spp. into (-)-hinokinin and
[38] O.A. Lameira, R.C.V.V. Martins-da-silva, M.D.G.B. Zoghbi, E.C.P.P. Oliveira, (-)-parabenzlactone, and their evaluation against oral pathogenic bacteria, Nat.
E.A. Oriental, M. Gra, B. Zoghbi, C. De Botminica, M. Paraense, E. Goeldi, Prod. Res. 6419 (2017) 1–14, [Link]
E.C.P.P. Oliveira, Seasonal variation in the volatiles of Copaifera duckei Dwyer [59] L.A.F. Paiva, K.M. de Alencar Cunha, F.A. Santos, N.V. Gramosa, E.R. Silveira,
growing wild in the State of Pará—Brazil, J. Essent. Oil Res. 21 (2009) 105–107, V.S.N. Rao, Investigation on the wound healing activity of oleo-resin from
[Link] Copaifera langsdorffii in rats, Phyther. Res. 16 (2002) 737–739, [Link]
[39] P.A. De Souza, L.P. Rangel, S.S. Oigman, M.M. Elias, A. Ferreira-Pereira, N.C. De 1002/ptr.1049.
Lucas, G.G. Leitão, Isolation of two bioactive diterpenic acids from Copaifera gly- [60] D. Masson-Meyers, T. Andrade, S. Leite, M. Frade, Cytotoxicity and wound healing
cycarpa oleoresin by high-speed counter-current chromatography, Phytochem. properties of Copaifera langsdorffii oleoresin in rabbits, Int. J. Nat. Prod. Sci. 3
Anal. 21 (2010) 539–543, [Link] (2013) 10–20.
[40] B.M. Tincusi, I.A. Jiménez, I.L. Bazzocchi, L.M. Moujir, Z.A. Mamani, J.P. Barroso, [61] J.J. de Lima Silva, S.B. Guimarães, E.R. da Silveira, P.R.L. de Vasconcelos,
A.G. Ravelo, B.V. Hernández, Antimicrobial terpenoids from the oleoresin of the G.G. Lima, S.M. Torres, R.C. de Vasconcelos, Effects of Copaifera langsdorffii Desf.
peruvian medicinal plant Copaifera paupera, Planta Med. 68 (2002) 808–812, on ischemia-reperfusion of randomized skin flaps in rats, Aesthetic Plast. Surg. 33
[Link] (2009) 104–109, [Link]
[41] M.D.G.B. Zoghbi, R.C.V. Martins-da-Silva, J.R. Trigo, Volatiles of oleoresins of [62] V.P. Ribeiro, C. Arruda, M. Abd El-Salam, J.K. Bastos, Brazilian medicinal plants
Copaifera paupera (Herzog) Dwyer C. Piresii Dwyer and C. Pubiflora Benth. with corroborated anti-inflammatory activities: a review, Pharm. Biol. 56 (2018)
(Leguminosae), J. Essent. Oil Res. 21 (2009) 403–404, [Link] 253–268, [Link]
10412905.2009.9700203. [63] F.B. Teixeira, R. de Brito Silva, O.A. Lameira, L.P. Webber, R.S. D’Almeida Couto,
[42] N.M. Gomes, C.M. Rezende, S.P. Fontes, M.E. Matheus, P.D. Fernandes, M.D. Martins, R.R. Lima, Copaiba oil-resin (Copaifera reticulata Ducke) modulates
Antinociceptive activity of Amazonian Copaiba oils, J. Ethnopharmacol. 109 the inflammation in a model of injury to rats’ tongues, BMC Complement. Altern.
(2007) 486–492, [Link] Med. 17 (2017) 313, [Link]
[43] C.G. Sachetti, R.R. de Carvalho, F.J.R.R. Paumgartten, O.A. Lameira, E.D. Caldas, [64] V.F. Veiga, L. Zunino, M.L. Patitucci, A.C. Pinto, J.B. Calixto, The inhibition of paw
Developmental toxicity of copaiba tree (Copaifera reticulata Ducke, Fabaceae) oedema formation caused by the oil of Copaifera multijuga Hayne and its fractions,
oleoresin in rat, Food Chem. Toxicol. 49 (2011) 1080–1085, [Link] J. Pharm. Pharmacol. 58 (2006) 1405–1410, [Link]
1016/[Link].2011.01.015. 7158.2006.tb01659.x.
[44] D.K.R. Bardají, J.J.M. da Silva, T.C. Bianchi, D. de Souza Eugênio, P.F. de Oliveira, [65] S. Baylac, Inhibition of 5-lipoxygenase by essential oils and other natural fragrant
L.F. Leandro, H.L.G. Rogez, R.C.S. Venezianni, S.R. Ambrosio, D.C. Tavares, extracts, Int. J. Aromather. 13 (2003) 138–142, [Link]
J.K. Bastos, C.H.G. Martins, Copaifera reticulata oleoresin: chemical characteriza- 4562(03)00083-3.
tion and antibacterial properties against oral pathogens, Anaerobe 40 (2016) [66] F. Hadji-Minaglou, O. Bolcato, The potential role of specific essential oils in the
18–27, [Link] replacement of dermacorticoid drugs (strong, medium and weak) in the treatment
[45] M.D.G.B. Zoghbi, E.H.A. Andrade, R.C.V. Martins-da-Silva, J.R. Trigo, Chemical of acute dry or weeping dermatitis, Int. J. Aromather. 15 (2005) 66–73, https://
variation in the volatiles of Copaifera reticulata Ducke (Leguminosae) Growing [Link]/10.1016/[Link].2005.03.013.
wild in the States of Pará and Amapá, Brazil, J. Essent. Oil Res. 21 (2009) [67] C. Kobayashi, T.O. Fontanive, B.G. Enzweiler, L.R. de Bona, T. Massoni, M.A. Apel,
501–503, [Link] A.T. Henriques, M.F. Richter, P. Ardenghi, E.S. Suyenaga, Pharmacological eva-
[46] G.S.C.J. Desmarchelier, J.M. Bustamante, R.R. Gil, J.D. Coussio, G.N. Ciccia, luation of Copaifera multijuga oil in rats, Pharm. Biol. 49 (2011) 306–313, https://
Profisetinidin type tannins responsible for antioxidant activity in Copaifera re- [Link]/10.3109/13880209.2010.515595.
ticulata, Pharmazie 56 (2001) 573–577. [68] F.D.S. Vargas, P.D.O. De Almeida, E.S.P. Aranha, A.P.D.A. Boleti, P. Newton,
[47] V.F. Veiga, A.C. Pinto, H.C. de Lima, The essential oil composition of Copaifera M.C. De Vasconcellos, V.F. Veiga, E.S. Lima, Biological activities and cytotoxicity
trapezifolia Hayne leaves, J. Essent. Oil Res. 18 (2006) 430–431, [Link] of diterpenes from Copaifera spp. oleoresins, Molecules 20 (2015) 6194–6210,
10.1080/10412905.2006.9699132. [Link]
[48] B. Adzu, S.O. Balogun, E. Pavan, S.D. Ascêncio, I.M. Soares, R.W.S. Aguiar, [69] R.A. Furtado, C.T.V. Bernardes, M.N. Da Silva, K.F. Zoccal, L.H. Faccioli,
R.V. Ribeiro, Â.M.S.E.S.S. Beserra, R.G. De Oliveira, L.I. da Silva, A.S. Damazo, J.K. Bastos, Antiedematogenic evaluation of Copaifera langsdorffii leaves hydro-
D.T.D.O. Martins, Evaluation of the safety, gastroprotective activity and me- ethanolic extract and its major compounds, Biomed. Res. Int. 2015 (2015) 1–7,
chanism of action of standardised leaves infusion extract of Copaifera malmei [Link]
harms, J. Ethnopharmacol. 175 (2015) 378–389, [Link] [70] N. de M. Gomes, C.M. de Rezende, S.P. Fontes, M.E. Matheus, A. da C. Pinto,
2015.09.027. P.D. Fernandes, Characterization of the antinociceptive and anti-inflammatory
[49] F. Chen, H. Al-Ahmad, B. Joyce, N. Zhao, T.G. Köllner, J. Degenhardt, activities of fractions obtained from Copaifera multijuga Hayne, J. Ethnopharmacol.
C.N. Stewart, Within-plant distribution and emission of sesquiterpenes from 128 (2010) 177–183, [Link]
Copaifera officinalis, Plant Physiol. Biochem. 47 (2009) 1017–1023, [Link] [71] C.H.G.G. Borges, M.G. Cruz, L.J. Carneiro, J.J.M.M. da Silva, J.K. Bastos,
org/10.1016/[Link].2009.07.005. D.C. Tavares, P.F. de Oliveira, V. Rodrigues, R.C.S.S. Veneziani, R.L.T.T. Parreira,
[50] C. de S. Galúcio, C.I. Benites, R.A.F. Rodrigues, M.R.W. Maciel, Sesquiterpenes G.F. Caramori, G.R. Nagurniak, L.G. Magalhães, S.R. Ambrósio, Copaifera duckei
recovery of copaiba oil-resin from molecular distillation, Quim. Nova 39 (2016) oleoresin and its main nonvolatile terpenes: in vitro schistosomicidal properties,
795–800, [Link] Chem. Biodivers. 13 (2016) 1348–1356, [Link]
[51] T. da S. Moraes, L.F. Leandro, L. de O. Silva, M.B. Santiago, A.B. Souza, 201600065.
R.A. Furtado, D.C. Tavares, R.C.S. Veneziani, S.R. Ambrósio, J.K. Bastos, [72] J.C. da Costa, G.M.R. Valladão, G. Pala, S.U. Gallani, S. Kotzent, A.E.M. Crotti,
C.H.G. Martins, In vitro evaluation of Copaifera oblongifolia oleoresin against L. Fracarolli, J.J.M. da Silva, F. Pilarski, Copaifera duckei oleoresin as a novel al-
bacteria causing oral infections and assessment of its xytotoxic potential, Curr. ternative for treatment of monogenean infections in pacu Piaractus mesopotamicus,
Pharm. Biotechnol. 17 (2016) 894–904, [Link] Aquaculture 471 (2017) 72–79.
1389201017666160415155359. [73] J. Lekana-Douki, S. Oyegue Liabagui, J. Bongui, R. Zatra, J. Lebibi, F.S. Toure-
[52] S.P. Arrhenius, J.H. Langenheim, Inhibitory effects of Hymenaea and Copaifera leaf Ndouo, In vitro antiplasmodial activity of crude extracts of Tetrapleura tetraptera
resins on the leaf fungus, Pestalotia subcuticularis, Biochem. Syst. Ecol. 11 (1983) and Copaifera religiosa, BMC Res. Notes 4 (2011) 506, [Link]
361–366, [Link] 1756-0500-4-506.
[53] W.F. de Moraes, P.M. Galdino, M.V.M. Nascimento, F.A. Vanderlinde, M.T.F. Bara, [74] G.A.G. de Souza, N.C. da Silva, J. de Souza, K.R.M. de Oliveira, A.L. da Fonseca,
E.A. Costa, J.R. de Paula, Triterpenes involved in the anti-inflammatory effect of L.C. Baratto, E.C.P. de Oliveira, F. de P. Varotti, W.P. Moraes, In vitro and in vivo
ethanolic extract of Pterodon emarginatus Vogel stem bark, J. Nat. Med. 66 (2012) antimalarial potential of oleoresin obtained from Copaifera reticulata Ducke
202–207, [Link] (Fabaceae) in the Brazilian Amazon rainforest, Phytomedicine 24 (2017) 111–118,
[54] K.B. Santiago, B.J. Conti, B.F.M.T. Andrade, J.J.M. da Silva, H.L.G. Rogez, [Link]
E.J. Crevelin, L.A. Beraldo de Moraes, R.C.S. Veneziani, S.R. Ambrósio, J.K. Bastos, [75] T.B. Le, C. Beaufay, D.T. Nghiem, M.-P. Mingeot-Leclercq, J. Quetin-Leclercq, In
J.M. Sforcin, Immunomodulatory action of Copaifera spp oleoresins on cytokine vitro anti-leishmanial activity of essential oils extracted from Vietnamese plants,
production by human monocytes, Biomed. Pharmacother. 70 (2015) 12–18, Molecules 22 (2017) 1071, [Link]
[Link] [76] F. de F. Fernandes, E. de P.S. Freitas, Acaricidal activity of an oleoresinous extract
[55] C.S. Mizuno, A.B. Souza, B.L. Tekwani, S.R. Ambrosio, R.C.S. Veneziani, Synthesis from Copaifera reticulata (Leguminosae: Caesalpinioideae) against larvae of the
and biological evaluation of polyalthic acid derivatives for the treatment of ne- southern cattle tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), Vet.
glected diseases, Bioorg. Med. Chem. Lett. 25 (2015) 5529–5531, [Link] Parasitol. 147 (2007) 150–154, [Link]
10.1016/[Link].2015.10.062. [77] A. Volpato, R.K. Grosskopf, R.C. Santos, R.A. Vaucher, R.P. Raffin, A.A. Boligon,
[56] A.O. dos Santos, E. Izumi, T. Ueda-Nakamura, B.P. Dias-Filho, V.F. da Veiga- M.L. Athayde, L.M. Stefani, A.S. Da Silva, R.K. Grosskopf, R.C. Santos,

18
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

R.A. Vaucher, R.P. Raffin, A.A. Boligon, M.L. Athayde, L.M. Stefani, A.S. Da Silva, Copaifera langsdorffii on rat uterus in vitro, Phyther. Res. 17 (2003) 320–324,
Influence of rosemary, andiroba and copaiba essential oils on different stages of [Link]
the biological cycle of the tick Rhipicephalus microplus in vitro, J. Essent. Oil Res. [96] L.A. Paiva, L. Gurgel, A. Campos, E. Silveira, V.S. Rao, Attenuation of ischemia/
27 (2015) 244–250, [Link] reperfusion-induced intestinal injury by oleo-resin from Copaifera langsdorffii in
[78] A.O. dos Santos, M.A. Costa, T. Ueda-Nakamura, B.P. Dias-Filho, V.F. da Veiga- rats, Life Sci. 75 (2004) 1979–1987, [Link]
Júnior, M.M. de Souza Lima, C.V. Nakamura, A.O. dos Santos, M.A. Costa, [97] A.G. da Silva, P. de F. Puziol, R. nunes Leitão, T.R. Gomes, R. Scherer,
T. Ueda-Nakamura, B.P. Dias-Filho, V.F. da Veiga-Júnior, M.M. de Souza Lima, M.L. Martins, Á.S. Cavalcanti, C.L. Carlos, Application of the essential oil from
C.V. Nakamura, Leishmania amazonensis: effects of oral treatment with copaiba oil copaiba (Copaifera langsdorffii Desf.) for acne vulgaris: a double-blind, placebo
in mice, Exp. Parasitol. 129 (2011) 145–151, [Link] controlled clinical trial, Altern. Med. Rev. 17 (2012) 69–75.
2011.06.016. [98] R.B. De Oliveira, E.B. Coelho, M.R. Rodrigues, A.R.D.M. Costa-Machado, J.P.B. De
[79] [Link] dos Santos, C.L. Donnici, E.R. d. S. Camargos, [Link] de Rezende, Sousa, A.A. Berretta, J.K. Bastos, Effect of the Copaifera langsdorffii Desf. leaf ex-
E.H. d. A. Andrade, L.A.L. Soares, L. d. M. Farias, [Link] de Carvalho, M. d. G. tract on the ethylene glycol-induced nephrolithiasis in rats, Evid. Complement.
Almeida, Effects of Copaifera duckei Dwyer oleoresin on the cell wall and cell di- Altern. Med. 2013 (2013) 1–10, [Link]
vision of Bacillus cereus, J. Med. Microbiol. 62 (2013) 1032–1037, [Link] [99] M.R.R. Tappin, J.F.G. Pereira, L.A. Lima, A.C. Siani, J.L. Mazzei, M.F.S. Ramos,
10.1099/jmm.0.060046-0. Análise química quantitativa para a padronização do óleo de copaíba por cro-
[80] S. Carbajosa, H.O. Rodríguez-Angulo, S. Gea, C. Chillón-Marinas, C. Poveda, matografia em fase gasosa de alta resolução, Quim. Nova 27 (2004) 236–240,
M.C. Maza, D. Colombet, M. Fresno, N. Gironès, L-arginine supplementation re- [Link]
duces mortality and improves disease outcome in mice infected with Trypanosoma [100] M.W. Biavattil, M. da PL, Daniela Dossin, Francisco C. Deschamps, Análise de
cruzi, PLoS Negl. Trop. Dis. 12 (2018), [Link] óleos-resinas de copaíba: contribuição para o seu controle de qualidade, Braz. J.
0006179 e0006179. Pharmacogn. 16 (2006) 230–235.
[81] E. Izumi, T. Ueda-Nakamura, V. Veiga-Júnior, C. Nakamura, Toxicity of oleoresins [101] A.F.F. de Vasconcelos, O.E.S. Godinho, Uso de métodos analíticos convencionados
from the genus Copaifera in Trypanosoma cruzi: a comparative study, Planta Med. no estudo da autenticidade do óleo de Copaíba, Quim. Nova 25 (2002)
79 (2013) 952–958, [Link] 1057–1060.
[82] F.D.S. Dorneles, D. Schafer, Aleksandro Silva, C.B. Oliveira, C.E.P. Zimmermann, [102] K.D.S. Barbosa, M. Yoshida, V.V. Scudeller, Detection of adulterated copaiba
L.D. Rosa, A.A. Tonin, E.C.P. De Oliveira, J.M. Santurio, S.G. Monteiro, (Copaifera multijuga Hayne) oil-resins by refractive index and thin layer chroma-
Susceptibility of Trypanosoma evansi in the copaiba oil: in vitro test and in mice tography, Rev. Bras. Farmacogn. 19 (2009) 57–60, [Link]
experimentally infected with the parasite, Acta Sci. Vet. 41 (2013) 1136. S0102-695X2009000100013.
[83] M.D. Baldissera, C.B. Oliveira, A.A. Tonin, P. Wolkmer, S.T.A. Lopes, R. Fighera, [103] P.C. Souza Barbosa, L.S. Moreira Wiedemann, R. da Silva Medeiros, P. de Tarso
M.M. Flores, E.C.P. Oliveira, R.C.V. Santos, A.A. Boligon, M.L. Athayde, Barbosa Sampaio, G. Vieira, V. Florêncio da Veiga-Junior, Phytochemical finger-
S.G. Monteiro, A.S. Da Silva, Toxic effect of essential oils (Copaifera spp) in the prints of copaiba oils (Copaifera multijuga Hayne) determined by multivariate
treatment of mice experimentally infected with Trypanosoma evansi, Biomed. Prev. analysis, Chem. Biodivers. 10 (2013) 1350–1360, [Link]
Nutr. 4 (2014) 319–324, [Link] 201200356.
[84] D. Kian, C.A.C. Lancheros, J.P. Assolini, N.S. Arakawa, V.F. Veiga-Júnior, [104] Y.F. Wong, T.M. Uekane, C.M. Rezende, H.R. Bizzo, P.J. Marriott, Qualitative
C.V. Nakamura, P. Pinge-Filho, I. Conchon-Costa, W.R. Pavanelli, S.F. Yamada- analysis of Copaifera oleoresin using comprehensive two-dimensional gas chro-
Ogatta, L.M. Yamauchi, Trypanocidal activity of copaiba oil and kaurenoic acid matography and gas chromatography with classical and cold electron ionisation
does not depend on macrophage killing machinery, Biomed. Pharmacother. 103 mass spectrometry, J. Chromatogr. A 1477 (2016) 91–99, [Link]
(2018) 1294–1301, [Link] 1016/[Link].2016.11.038.
[85] N. de M. Gomes, C. de M. Rezende, S.P. Fontes, A.M.C. Hovell, R.G. Landgraf, [105] J.J.M. da Silva, E.J. Crevelin, L.J. Carneiro, H. Rogez, R.C.S. Veneziani,
M.E. Matheus, A. da C. Pinto, P.D. Fernandes, Antineoplasic activity of Copaifera S.R. Ambrósio, L.A. Beraldo Moraes, J.K. Bastos, Development of a validated ultra-
multijuga oil and fractions against ascitic and solid Ehrlich tumor, J. high-performance liquid chromatography tandem mass spectrometry method for
Ethnopharmacol. 119 (2008) 179–184, [Link] determination of acid diterpenes in Copaifera oleoresins, J. Chromatogr. A 1515
033. (2017) 81–90, [Link]
[86] H.M. dos Santos Júnior, D.F. Oliveira, D.A. de Carvalho, J.M.A. Pinto, [106] J. de Sousa, A. Brancalion, M. Júnior, J.K. Bastos, A validated chromatographic
V.A.C. Campos, A.R.B. Mourão, C. Pessoa, M.O. de Moraes, L.V. Costa-Lotufo, method for the determination of flavonoids in Copaifera langsdorffii by HPLC, Nat.
Evaluation of native and exotic Brazilian plants for anticancer activity, J. Nat. Prod. Commun. 7 (2012) 25–28 (accessed January 2, 2017), [Link]
Med. 64 (2010) 231–238, [Link] [Link]/pubmed/22428235.
[87] J.M. Senedese, J.M. Alves, I.M. de S. Lima, E.A.P. de Andrade, R.A. Furtado, [107] A.B. Souza, M.R. Moreira, C.H.G. Borges, M.R. Simão, J.K. Bastos, J.P.B. de Sousa,
J.K. Bastos, D.C. Tavares, Chemopreventive effect of Copaifera langsdorffii leaves S.R. Ambrosio, R.C.S. Veneziani, J.P.B. Sousa, S.R. Ambrosio, R.C.S. Veneziani,
hydroalcoholic extract on 1,2-dimethylhydrazine-induced DNA damage and pre- Development and validation of a rapid RP-HPLC method for analysis of (−)-co-
neoplastic lesions in rat colon, BMC Complement. Altern. Med. 13 (2013) 3, palic acid in copaíba oleoresin, Biomed. Chromatogr. 27 (2012) 280–283, https://
[Link] [Link]/10.1002/bmc.2788.
[88] A. Ohsaki, L.T. Yana, S. Itoa, H. Edatsugib, D. Iwatab, Y. Komodaa, B. Science, The [108] V.R. De Almeida Borges, A.F. Ribeiro, C. De Souza Anselmo, L.M. Cabral, V.P. De
isolation and in vivo potent antitumor activity of clerodane diterpenoid from the Sousa, Development of a high performance liquid chromatography method for
oleoresin of the Brazilian medicinal plant, Copaifera langsdorffii Desfon, Bioorg. quantification of isomers β-caryophyllene and α-humulene in copaiba oleoresin
Med. Chem. Lett. 4 (1994) 2889–2892, [Link] using the Box-Behnken design, J. Chromatogr. B: Anal. Technol. Biomed. Life Sci.
80834-9. 940 (2013) 35–41, [Link]
[89] L.V. Costa-Lotufo, G.M.A. Cunha, P.A.M. Farias, G.S.B. Viana, K.M.A. Cunha, [109] D. de O. Dias, M. Colombo, R.G. Kelmann, T.P. De Souza, V.L. Bassani,
C. Pessoa, M.O. Moraes, E.R. Silveira, N.V. Gramosa, V.S.N. Rao, The cytotoxic and H.F. Teixeira, V.F. Veiga, R.P. Limberger, L.S. Koester, Optimization of headspace
embryotoxic effects of kaurenoic acid, a diterpene isolated from Copaifera langs- solid-phase microextraction for analysis of β-caryophyllene in a nanoemulsion
dorffii oleo-resin, Toxicon 40 (2002) 1231–1234, [Link] dosage form prepared with copaiba (Copaifera multijuga Hayne) oil, Anal. Chim.
0101(02)00128-9. Acta 721 (2012) 79–84, [Link]
[90] L.A. Paiva, V.S. Rao, N. Gramosa, E. Silveira, Gastroprotective effect of Copaifera [110] L.G. Lucca, S.P. de Matos, B.T. Borille, D. de O. Dias, H.F. Teixeira, V.F. Veiga,
langsdorffii oleo-resin on experimental gastric ulcer models in rats, J. R.P. Limberger, L.S. Koester, Determination of β-caryophyllene skin permeation/
Ethnopharmacol. 62 (1998) 73–78, [Link] retention from crude copaiba oil (Copaifera multijuga Hayne) and respective oil-
00058-0. based nanoemulsion using a novel HS-GC/MS method, J. Pharm. Biomed. Anal.
[91] F.T. Tavares Trindade, R.G. Stabeli, A.A. Pereira, V.A. Facundo, A. de Almeida e 104 (2015) 144–148, [Link]
Silva, Copaifera multijuga ethanolic extracts, oilresin, and its derivatives display [111] J. Muller, R. Fernandes, M. Batista, E. Moura, R. Silva, E. Ferreira-Filho,
larvicidal activity against Anopheles darlingi and Aedes aegypti (Diptera: Culicidae), R. Oliveira, M. Fernandes, Studies of Copaifera luetzelburgiiHarms in reproductive
Rev. Bras. Farmacogn. 23 (2013) 464–470, [Link] pharmacology: in vivo and in vitro approaches, Afr. J. Biotechnol. 12 (2013)
695X2013005000038. 3864–3871, [Link]
[92] F.A.C. de Mendonça, K.F.S. da Silva, K.K. dos Santos, K.A.L. Ribeiro Jr, [112] E.S. Gonçalves, J.R. Silva, C.L. Gomes, M.B.L. Nery, D.M. a. F. Navarro,
A.E.G. Sant’Ana, Activities of some Brazilian plants against larvae of the mosquito G.K.N. Santos, J.C. Silva-Neto, J.H. Costa-Silva, A.V. Araújo, A.G. Wanderley,
Aedes aegypti, Fitoterapia 76 (2005) 629–636, [Link] Effects of the oral treatment with Copaifera multijuga oil on reproductive perfor-
2005.06.013. mance of male Wistar rats, Rev. Bras. Farmacogn. 24 (2014) 355–362, [Link]
[93] D.S. Alves, G.A. Carvalho, D.F. Oliveria, R.R. Sâmia, M.A. Villas-Boas, org/10.1016/[Link].2014.07.014.
G.A. Carvalho, A.D. Côrrea, Toxicity of copaiba extracts to armyworm (Spodoptera [113] C.S. Lima, B.J.L.L. De Medeiros, H.A.S.S. Favacho, K.C. Dos Santos, B.R. De
frugiperda), Afr. J. Biotechnol. 11 (2012) 6578–6591, [Link] Oliveira, J.C. Taglialegna, E.V.M.M. Da Costa, K.J. De Campos, J.C.T.T. Carvalho,
AJB11.196. Pre-clinical validation of a vaginal cream containing copaiba oil (reproductive
[94] [Link] da Silva, V.R.D.A. Borges, L.D.C.B. Pereira, R. Ferrari, R.M. de Mattos, toxicology study), Phytomedicine 18 (2011) 1013–1023, [Link]
E.G.D.O. Barros, C.Y. Palmero, P.D. Fernandes, P.R. de Carvalho, V. Pereira de 1016/[Link].2011.05.004.
Sousa, L.M. Cabral, L.E. Nasciutti, The oil-resin of the tropical rainforest tree [114] M.R. Almeida, J.D.A.C. Darin, L.C. Hernandes, M.F. de Souza Ramos,
Copaifera langsdorffii reduces cell viability, changes cell morphology and induces L.M.G. Antunes, O. de Freitas, Genotoxicity assessment of Copaiba oil and its
cell death in human endometriotic stromal cultures, J. Pharm. Pharmacol. 67 fractions in Swiss mice, Genet. Mol. Biol. 35 (2012) 664–672, [Link]
(2015) 1744–1755, [Link] 1590/S1415-47572012005000052.
[95] K.M. de Alencar Cunha, L.A.F. Paiva, F.A. Santos, N.V. Gramosa, E.R. Silveira, [115] O. Castro-e-Silva, F. Ramalho, L. Ramalho, S. Zucoloto, M.E. Souza, M. Brito,
V.S.N. Rao, Smooth muscle relaxant effect of kaurenoic acid, a diterpene from J. Reis, A. Bastos, Anti-proliferative activity of oleoresin from Brazilian Copaifera

19
C. Arruda et al. Biomedicine & Pharmacotherapy 109 (2019) 1–20

on liver regeneration in rats, J. Hepatol. 18 (2004) 92–94, [Link] [132] J. Nascimento, J.H. Langenheim, Leaf sesquiterpenes and phenolics in Copaifera
1016/S0168-8278(02)80779-6. multijuga on contrasting soil types in a central amazonian rain forest, Biochem.
[116] J.M. Alves, C.C. Munari, M. de Azevedo Bentes Monteiro Neto, R.A. Furtado, Syst. Ecol. 14 (1986) 615–624, [Link]
J.M. Senedese, J.K. Bastos, D.C. Tavares, In vivo protective effect of Copaifera [133] V.F. Veiga, A.C. Pinto, H.C. de Lima, The essential oil composition of Copaifera
langsdorffii hydroalcoholic extract on micronuclei induction by doxorubicin, J. trapezifolia Hayne leaves, J. Essent. Oil Res. 18 (2006) 430–431, [Link]
Appl. Toxicol. 33 (2013) 854–860, [Link] 10.1080/10412905.2006.9699132.
[117] A. Oliveira dos Santos, T. Ueda-Nakamura, B. Prado Dias Filho, V. Florêncio da [134] L.C. Branch, M.F. da Silva, Folk medicine of alter of Alter do Chão, Pará, Brazil,
Veiga Junior, Â. da Cunha Pinto, C. Vataru Nakamura, Antimicrobial activity of Acta Amaz. 13 (1983) 737–797, [Link]
copaiba oils obtained from different species of Copaifera in Brazil, Planta Med. 73 [135] J.H.G. Lago, J. Tezoto, P.B. Yazbek, F. Cassas, J. de F.L. Santos, E. Rodrigues,
(2007) 277–281, [Link] Exudates used as medicine by the “caboclos river-dwellers” of the Unini River,
[118] R. Fenner, A.H. Betti, L.A. Mentz, S.M.K. Rates, Plantas utilizadas na medicina AM, Brazil – classification based in their chemical composition, Rev. Bras.
popular brasileira com potencial atividade antifúngica, Rev. Bras. Ciênc. Farm. 42 Farmacogn. 26 (2016) 379–384, [Link]
(2006) 369–394, [Link] [136] R. de Almeida Vaucher, J.L. Giongo, L.P. Bolzan, M.S. Côrrea, V.P. Fausto, C.F. dos
[119] J.C.T. Carvalho, V. Cascon, L.S. Possebon, M.S.S. Morimoto, L.G.V. Cardoso, S. Alves, L.Q.S. Lopes, A.A. Boligon, M.L. Athayde, A.P. Moreira, A. Brandelli,
M.A.C. Kaplan, B. Gilbert, Topical antiinflammatory and analgesic activities of R.P. Raffin, R.C.V. Santos, Antimicrobial activity of nanostructured Amazonian
Copaifera duckei Dwyer, Phyther. Res. 19 (2005) 946–950, [Link] oils against Paenibacillus species and their toxicity on larvae and adult worker
1002/ptr.1762. bees, J. Asia Pac. Entomol. 18 (2015) 205–210, [Link]
[120] V.R. De Almeida Borges, J.H. Da Silva, S.S. Barbosa, L.E. Nasciutti, L.M. Cabral, 2015.01.004.
V.P. De Sousa, Development and pharmacological evaluation of in vitro nano- [137] B.F. Murbach Teles Andrade, L. Nunes Barbosa, I. da Silva Probst, A. Fernandes
carriers composed of lamellar silicates containing copaiba oil-resin for treatment Júnior, Antimicrobial activity of essential oils, J. Essent. Oil Res. 26 (2014) 34–40,
of endometriosis, Mater. Sci. Eng. C 64 (2016) 310–317, [Link] [Link]
[Link].2016.03.094. [138] P.L. Tobouti, M.C.M. Mussi, D.C.P. Rossi, F.M. Pigatti, C.P. Taborda, L.A. de Assis
[121] E. Rodrigues, J.M. Duarte-Almeida, J.M. Pires, Perfil farmacológico e fitoquímico Taveira, S.C.O.M. de Sousa, Influence of melaleuca and copaiba oils on Candida
de plantas indicadas pelos caboclos do parque nacional do jaú (am) como po- albicans adhesion, Gerodontology 33 (2016) 380–385, [Link]
tenciais analgésicas. parte i, Braz. J. Pharmacogn. 20 (2010) 981–991, [Link] ger.12172.
org/10.1590/S0102-695X2010005000008. [139] S. Baylac, P. Racine, Inhibition of human leukocyte elastase by natural fragrant
[122] B. Kumar, M. Vijayakumar, R. Govindarajan, P. Pushpangadan, extracts of aromatic plants, Int. J. Aromather. 14 (2004) 179–182, [Link]
Ethnopharmacological approaches to wound healing-exploring medicinal plants of org/10.1016/[Link].2004.09.008.
India, J. Ethnopharmacol. 114 (2007) 103–113, [Link] [140] M.D.G.B. Zoghbi, E.H.a. Andrade, R.C.V. Martins-da-Silva, J.R. Trigo, Ducke
2007.08.010. (Leguminosae) growing wild in the states of Pará and Amapá, Brazil, J. Essent. Oil
[123] B.C. Cavalcanti, L.V. Costa-Lotufo, M.O. Moraes, R.R. Burbano, E.R. Silveira, Res. 21 (2009) 501–503, [Link]
K.M.A. Cunha, V.S.N. Rao, D.J. Moura, R.M. Rosa, J.A.P. Henriques, C. Pessoa, [141] R. Geris, I.G. Da Silva, H.H.G. Da Silva, A. Barison, E. Rodrigues-Filho,
Genotoxicity evaluation of kaurenoic acid, a bioactive diterpenoid present in A.G. Ferreira, Diterpenoids from Copaifera reticulata ducke with larvicidal activity
Copaiba oil, Food Chem. Toxicol. 44 (2006) 388–392, [Link] against Aedes aegypti (L.) (Diptera, Culicidae), Rev. Inst. Med. Trop. Sao Paulo 50
fct.2005.08.011. (2008) 25–28, [Link]
[124] K.M.A. Cunha, E.R. Silveira, F.A. Santos, V.S. Rao, Effects of kaurenoic acid, a [142] Z. Hajdu, J. Hohmann, An ethnopharmacological survey of the traditional medi-
bioactive diterpene on embryo implantation and pregnancy outcome in mice, cine utilized in the community of Porvenir, Bajo Paraguá Indian Reservation,
Curr. Top. Toxicol. 7 (2011) 89–94. Bolivia, J. Ethnopharmacol. 139 (2012) 838–857, [Link]
[125] A. Fonseca, F. Estrela, T. Moraes, L. Carneiro, J. Bastos, R. Santos, S. Ambrósio, 2011.12.029.
C. Martins, R. Veneziani, In vitro antimicrobial activity of plant-derived diterpenes [143] J.M. Alves, C.C. Munari, M. de Azevedo Bentes Monteiro Neto, R.A. Furtado,
against bovine mastitis bacteria, Molecules 18 (2013) 7865–7872, [Link] J.M. Senedese, J.K. Bastos, D.C. Tavares, In vivo protective effect of Copaifera
org/10.3390/molecules18077865. langsdorffii hydroalcoholic extract on micronuclei induction by doxorubicin, J.
[126] A.B. Souza, C.H.G. Martins, M.G.M. Souza, N.A.J.C. Furtado, V.C.G. Heleno, Appl. Toxicol. 33 (2013) 854–860, [Link]
J.P.B. de Sousa, E.M.P. Rocha, J.K. Bastos, W.R. Cunha, R.C.S. Veneziani, [144] A.R.M. Costa-Machado, J.K. Bastos, L.A.P. de Freitas, Dynamic maceration of
S.R. Ambrósio, Antimicrobial activity of terpenoids from Copaifera langsdorffii Copaifera langsdorffi leaves: a technological study using fractional factorial design,
Desf. against cariogenic bacteria, Phyther. Res. 25 (2010) 215–220, [Link] Rev. Bras. Farmacogn. 23 (2013) 79–85, [Link]
org/10.1002/ptr.3244. 695X2012005000116.
[127] D. Zimmermam-Franco, E. Bolutari, H. Polonini, A. do Carmo, M. das Graças, [145] A.R.M. Costa, L.A.P. Freitas, J. Mendiola, E. Ibáñez, Copaifera langsdorffii super-
A.M. Chaves, N. Raposo, Antifungal activity of Copaifera langsdorffii Desf pleoresin critical fluid extraction: chemical and functional characterization by LC/MS and in
against dermatophytes, Molecules 18 (2013) 12561–12570, [Link] vitro assays, J. Supercrit. Fluids 100 (2015) 86–96, [Link]
3390/molecules181012561. supflu.2015.02.028.
[128] A.G. Barreto Júnior, E.C. Biscaia Junior, V.F. da Veiga Junior, A.C. Pinto, S.F. de [146] J.H. Langenheim, C.L. Convis, C.A. Macedo, W.H. Stubblebine, Hymenaea and
Carvalhaes, M.A.M. Maciel, Cromatografia de troca-iônica aplicada ao isolamento Copaifera leaf sesquiterpenes in relation to lepidopteran herbivory in southeastern
da fração ácida do óleo de copaíba (Copaifera multijuga) e da sacaca (Croton ca- Brazil, Biochem. Syst. Ecol. 14 (1986) 41–49, [Link]
jucara), Quim. Nova 28 (2005) 719–722, [Link] 1978(86)90084-0.
40422005000400028. [147] S.P. Arrhenius, C.E. Foster, C.G. Edmonds, J.H. Langenheim, Sesquiterpenes in leaf
[129] S.R.M. Lima, V.F.V. Junior, H.B. Christo, A.C. Pinto, P.D. Fernandes, In vivo and in pocket resins of Copaifera species, Phytochemistry 22 (1983) 471–472, https://
vitro studies on the anticancer activity of Copaifera multijuga hayne and its frac- [Link]/10.1016/0031-9422(83)83026-X.
tions, Phyther. Res. 17 (2003) 1048–1053, [Link] [148] J.J. Lafont, M.S. Páez, E. Lans, Composición fisicoquímica de la semilla y del aceite
[130] R.J.A. Deus, A.S.C. Carvalho, D.A.D.S. Banna, M.S. Arruda, C.N. Alves, A.S. Santos, de la semilla del Canime (Copaifera officinalis L), Inf. Tecnol. 22 (2011) 19–26,
In vitro fungitoxic effect of the oil resin and the essential oil of copaiba (Copaifera [Link]
multijuga Hayne), Rev. Bras. Plantas Med. 11 (2009) 347–353. [149] V.F. Veiga Junior, M.A. Andrade Junior, I.D.K. Ferraz, H.B. Christo, A.C. Pinto,
[131] C.L. Morelli, M. Mahrous, M.N. Belgacem, M.C. Branciforti, R.E.S. Bretas, J. Bras, Constituintes das sementes de Copaifera officinalis L, Acta Amaz. 37 (2007)
Natural copaiba oil as antibacterial agent for bio-based active packaging, Ind. 123–126, [Link]
Crops Prod. 70 (2015) 134–141, [Link]

20