Technological prospection of the biological activities of Plectranthus amboinicus (Loureiro) Sprengel: an integrative review Prospecção tecnológica das atividades biológicas de Plectranthus amboinicus (Loureiro) Sprengel: uma revisão integrativa Prospección tecnológica de las actividades biológicas de Plectranthus amboinicus (Loureiro) Sprengel: una revisión integradora

Plectranthus amboinicus (Loureiro) Sprengel is popularly known as malvariço or indian borage. This paper carries out the technological prospecting of this species with an emphasis on its biological activities. The research consisted of an integrative literature review based on information collected from patent and electronic databases. Initially, 3019 results were obtained, which, after the screening, selection and analysis process, resulted in 55 articles and 48 patent documents. It was found that 77.08% (n=37) of patent deposits refer to obtaining or improving the process and 22.92% (n=11) to the finished product related to the following indications: wound healing, mainly in diabetics, Research, Society and Development, v. 10, n. 5, e30110514653, 2021 (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i5.14653 2 followed by treatment of rheumatoid arthritis, hepatocellular carcinomas, breast cancer and melanoma, treatment for acne caused by P. acnes, S. aureus and P. aeruginosa, oral cavity cleansing agent, treatment for osteoporosis, nutraceutical composition for lactation and insect repellent. In the databases of scientific articles, 55 documents were selected, proving the following actions: antimicrobial, antioxidant, anticarcinogenic, anti-inflammatory, diuretic, nephroprotective, larvicide, analgesic, veterinary food additive, anti-leishmania, antipsoriasis, antidiarrheal, antidiabetic, hepatoprotective and anthelmintic. The scientific studies analyzed reaffirm other actions reported in popular use, but which are not described in the patent processes. Therefore, there are many proven biological properties of P. amboinicus that should be further explored and exploited in the pharmaceutical, cosmetic and food sectors in order to generate new products and patents.


Introduction
Biotechnology research has evolved quickly and safely in the production of information, which can be seen by the growing development of new processes and products. Technological prospecting studies have shown a way of schematizing and informing technoscientific advances, which has narrowed the distance between invention and commercialization of products (Amparo et al., 2012;Caruso & Tigre, 2004;Oliveira & Quental, 2012).
The possibility of identifying and obtaining compounds with varied pharmacological actions has stimulated research on medicinal plants (Souto-Maior et al., 2011). Species of the genus Plectranthus (Lamiaceae) are found on four continents: Africa, America, Oceania, and Asia (Arumugam et al., 2016;Ruan et al., 2019).
Plectranthus amboinicus (Loureiro) Sprengel, popularly known as malvariço or indian borage, is a herbaceous plant, up to one meter long, succulent with a brittle stem, oval leaves with thick apex, jagged edges and a thick petiole with a sharp tip. Phytochemical studies have described the existence of 76 volatile and 30 non-volatile compounds that belong to different classes of constituents, such as monoterpenes, diterpenes, triterpenes, sesquiterpenes, phenolics, flavonoids, esters, alcohols and aldehydes, with diterpenoids, triterpenoids and essential oils (OEs) being the most common metabolites. Some noticeable pharmacological actions of this plant are: antimicrobial, anti-inflammatory, antitumor, healing, antiepileptic, and analgesic activities (Arumugam et al., 2016;Ruan et al., 2019).
The low cost and therapeutic benefits presented have led part of the population to use natural products. New products made from medicinal plants originated in popular knowledge and were later scientifically proven. The systematization of records of the use of this species aims to stimulate access to innovative research and the generation of patents through a selection of bioassays that exhibited biological effects in a simple, sensitive and safe way.
This research sought to identify scientific and technological advances in processes and/or products associated with biological properties, in order to create an updated information source that disseminates the evidence of the pharmacological action. The idea of conducting a logical and rational analysis of the existing data in patent databases and academic research reveals the current panorama of the world market and leads to a compendium of relevant information on the main lines of research developed with P. amboinicus.

Methodology
The research consisted of an integrative literature review that seeks the synthesis of knowledge and the incorporation of the applicability of the results of significant studies in practice (Silveira, 2005;Souza et al., 2010). It was based on information collected from databases of patents and scientific articles. The question that guided the study was: Are there scientifically proven therapeutic actions by Plectranthus amboinicus that have not yet been patented?

Data sources
Scientific research was carried out in four electronic databases: SCIELO (Scientific Electronic Library Online), Google Scholar, PUBMED (National Library of Medicine of the United States, National Institutes of Health) and CAPES Portal (Coordination for the Improvement of Higher Education Personnel). The technological prospection was also carried out on four patent databases: USPTO (United States Patent and Trademark Office), WIPO (World Intellectual Property Organization), ESPACENET, and INPI (National Institute of Industrial Property). The search was performed by combining two criteria: i) descriptors and ii) search for international patent classification -IPCcodes (prospecting). Scientific names were used for titles and abstracts, with or without the Boolean operator "AND". The descriptors used in the study are described in Figure 1.

Selection criteria
The selection criteria were based on the inclusion and non-inclusion criteria established by the researchers. Among the 3019 files found, 154 documents (83 articles and 71 patents) were selected for reading in full. The inclusion criterion covered materials that addressed: technological applications, therapeutic uses, formulations developed and the parts used of this species. The non-inclusion criteria were: year of publication of the articles, duplication in the patent databases and academic research consulted, articles and/or patents whose focus was not directed to biological uses and properties, development of formulation or technologies involving this species.
During the research with heteronyms of the species P. amboinicus (taken from the site: reflora.jbrj.gov.br) and the Boolean operator "AND" plus "Indian Borage", the files were selected and quantified, excluding those that did not correspond to the criteria. Review articles, conference abstracts and articles that studied their effects associated with other plants or other properties of the same were excluded. The selection and quantification of the data obtained in each analysis process can be seen in Figure 2.

Data analysis
Data collection and analysis was carried out in January 2020, through a selection of scientific articles published in the last 10 years (from January 2010 to December 2019) and all published patents found in the patent databases, without temporal (1976 to 2019) or language delimitations, to discuss and cross-check the data found. The list of relevant article references to the research was verified and, then, they were subjected to an exploratory and selective reading to identify data related to the proposed theme according to the objective of the study.
The first step was to analyze the titles and read the abstracts. The files that met the inclusion criteria were read in full and then selected according to the objective of this work. Some aspects that guided the analysis of the archives were the year of publication, research articles and essential observations on the proposed theme. Regarding the analysis of the patents, the criteria used to determine compliance with the research topic were: title, abstract, documentation and patent number. The files that had the scientific or popular name of the plant in the title and/or abstract were selected for further analysis of prospection by classification, year and country.
The extracted data were organized in a Microsoft® Excel spreadsheet. After corrections, the information was exported to the Statistical Package for the Social Sciences (SPSS) software, version 26.0, where statistical processing was performed. To characterize the sample, descriptive statistics were performed using simple frequency.

Results and Discussion
The data selection process (scientific articles and data from patent databases) initially generated 3019 files matching the chosen descriptors and study period, including files duplicated by the research sources. Of the 3019 files retrieved, 2692 Research, Society andDevelopment, v. 10, n. 5, e30110514653, 2021 (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i5.14653 6 results corresponded to scientific articles and 327 corresponded to results in patent databases. The quantification of these data can be observed in Table 1.
After selection and analysis (titles and abstracts), 83 scientific articles and 71 patents issued (154 documents) were chosen to be read in full. For a better selection of data, a search for therapeutic indications was carried out in patent database and academic research, with a subsequent reading in full of the selected results. After a critical review of the articles, 48 patent documents and 55 scientific publications were selected for further analysis and discussion.
The search for scientific articles in the chosen electronic databases provided 2692 initial results, most of them in Google Scholar (n = 1756), followed by the Portal of CAPES (n = 884), PUBMED (n = 30) and SCIELO (n = 22). The search in the patent databases returned 327 results, most of them found in WIPO (n = 149), followed by INPI (n = 110), ESPACENET (n = 42) and USPTO (n = 26).
After the initial results obtained from the patent databases (n = 327) were filtered by the inclusion / exclusion criteria, 57 issued patents were returned. It was observed, however, that 05 of the USPTO results and 03 of ESPACENET results were also filed at WIPO and 1 result was filed. After the final selection, 48 issued patents were chosen for final analysis, all extracted from WIPO.
The databases were searched and the collected date were crossed. Some differences were observed in the results obtained with the use of popular synonyms and their applications in the formulations developed in the selected works (Table   1). Table 1. Initial quantification of data obtained with the selected descriptors.
It was also detected in the prospecting research of this species a greater number of patents for isolated use classified in the A61K group (97.3%) and one in the A61P group (2.7%). Associated use patents are classified into the groups listed in Table 2. From the data presented above, it can be seen that this species has been more studied in relation to the area of the A61K group (preparations for medical, dental or hygienic use), totaling 36 patents for isolated use (97.3%). The A61K group appears in all 48 patents consulted when associated with other purposes (classifications), directing all studies to applications in humans, mainly for medical, dental or toilet purposes. The pharma industry's R&D (Research and Development) sector in recent decades has shown only slight growth in innovation, while inventions and innovations in medical technologies have advanced more rapidly (Dutta et al., 2019).
According to the data obtained, the patents are mainly process patents, for human health purposes. These patents refer mainly to anti-inflammatory, healing and anti-tumor actions, especially with new extraction methods or improvement methods with a predominance of process patents.

Prospecting in scientific article databases
The search in academic research databases returned 83 documents, of which 55 were selected after reading in full.

Antimicrobial and antidiarrheal activities
In twenty-two selected articles on antimicrobial activity, studies with extracts and OE extracted from the leaves were found. These studies confirmed its activity against gram negative bacteria:  The hydroalcoholic extract of dry leaves at concentrations of 50mg/ml and 100mg/ml was also able to inhibit the growth of Pseudomonas aeruginosa (Sreelakshmy & Thangapandiyan, 2019), Shigella bayedir (Ismayil & Nimila, 2019), Aspergillus niger, Bacillus subtilis and Staphylococcus aureus, and its effect was compared to that of the standard drugs tested: Ampicilline trihydrate and Amphoterecine B (Majee et al., 2013). In the study by Bhatt & Negi (2012), the acetonic extract of the leaves showed superior activity to the ethyl acetate extract against the pathogens of food origin: Bacillus cereus, Escherichia coli, Staphylococcus aureus and Yersinia enterocolitica.
In the study by Bhatt et al. (2013) with methanolic extract, the action of this species was proven against pathogens present in food infections such as: Y. enterocolitica, E. coli, S. aureus and B. cereus. Years later, Shubha & Bhatt (2015) published research on the prebiotic effect of the aqueous extract proving the ability of this extract to stimulate the growth of Lactobacillus plantarum. This probiotic bacterium consumed the constituents of the extract to produce metabolic enzymes, thus stimulating its growth. Still in this study, the extract inhibited the growth of pathogens (E. coli and Salmonella typhimurium), proving the popular use of this species as an antidiarrheal in foodborne infections.
Extracts and OE of this species were tested to analyse its viability as an anti-biofilm. There are studies that describe the property of nanoparticles, synthesized from the aqueous extract of its leaves treated with zinc nitrate solution, being able to inhibit biofilm formation of methicillin-resistant Staphylococcus aureus (Vijayakumar et al., 2015). In the study of efficacy against Streptococcus pyogenes biofilms, the methanolic extract of the leaves also showed inhibitory effect functioning as antibiofilm (Manimekalai et al., 2016;Zhang et al., 2017). The OE tested on oxacillin-and vancomycin-resistant S. aureus species also worked as a viable alternative against these biofilms (Vasconcelos et al., 2017).
The leaves of indian borage can be used to treat opportunistic infections in HIV-infected patients, as its OE has an effect against C. albicans and Cryptococcus neoformans (Asiimwe, 2015). There are also reports of OE activity against fungi of the genus Candida: C.albicans, C.parapsilosis and C.tropicalis (EL-Zefzafy et al., 2016;Manjamalai et al., 2012;Santos et al., 2016;Vijayakumar et al., 2015).

Antioxidant, anti-inflammatory and analgesic activities
In twelve articles, results showed antioxidant action, in vitro, of the leaves, roots and stems of this species. In most of these studies, the use of hydroalcoholic leaves extract showed better activity when compared to other types of solvents (Amarasiri et al., 2018;Bezerra et al., 2017;Bhatt & Negi, 2012;Bhatt et al., 2013;Bole & Kumudini, 2014;El-hawary et al., 2012;Haryani et al., 2018;Hasibuan et al., 2013;Patel et al., 2010a;Prasenjit et al., 2011). In the comparative study carried out by Bhatt & Negi (2012) with different types of solvents: hexane, acetone, ethyl acetate metanol and hydroalcohol, a greater antioxidant activity was detected in the ethyl acetate extract of the leaves, followed by the acetonic extract.
Seven articles reported anti-inflammatory activity, with aqueous, ethanolic extracts and/or OE. The anti-inflammatory activity of the ethanolic extract of the leaves proved to be excellent when compared to indomethacin and hydrocortisone, while the essential oil showed anti-inflammatory activity similar to diclofenac sodium (standard drugs tested) (Devi & Periyanayagam, 2010;El-hawary et al., 2012;Manjamalai et al., 2012).
There are records of the use of aqueous extract to inhibit pain induced by acetic acid and formalin and inflammation induced by carrageenan. It was found that this effect was related to the amount of carvacrol in the aqueous extract, resulting in the modulation of antioxidant enzymes in the liver, leading to a decrease in malondialdehyde, tumor necrosis factor alpha (TNF-α) and cyclooxygenase 2 (COX-2) production of paw edema in rats (Akinbo et al., 2018;Chiu et al., 2012;Leu et al., 2019). Thymoquinone was detected in the hexanic fraction of leaves as a compound that inhibits the expression of TNF-α (Chen et al., 2014).
Recent studies report that the constituents rosmarinic acid, cirsimaritin, salvigenin and carvacrol are correlated with the inhibition of inflammasome and activation of inflammatory responses, with an inhibitory activity observed in NLRP3 inflammasome blockade (Leu et al., 2019).
Three articles were found that prove analgesic activity: Chiu et al., 2012;El-Hawary et al., 2012 andMajee et al., 2013. In two studies, indomethacin was the standard drug used, and for El-Hawary et al. (2012), the best response was in alcoholic extracts and ethyl acetate fractions of the leaves while for Chiu et al. (2012) the best response was in the aqueous extract. In the study by Majee et al., 2013, using pentazocine as the standard drug, the analgesic effect was confirmed in the hydroalcoholic extract of the leaves. These records scientifically prove its popular use in pain relief.
In Bhatt et al. (2013) the sample extracted in methanol presented as constituents: total phenols, flavonoids and condensed tannins in greater proportions when compared to the others. Cytotoxic activity against cervical adenocarcinoma cells (HeLa) was detected in the hexane and alcoholic leaves extracts (Hasibuan et al., 2013;Hasibuan & Rosidah, 2015). In the study by Hasibuan et al. (2013) the ethyl acetate extract showed the highest content of phenolic compound and the best antioxidant and anticancer response.The ethanolic extract was transformed into nanoparticles by gelation, to test its cytotoxicity against HeLa, MCF-7 and T47D cells in vitro. There are also records of studies showing synergism with doxorubicin in ethyl acetate extract to inhibit metastasis in T47D breast cancer cells Rosidah & Hasibuan, 2014).
There are also reports of the anticarcinogenic potential, with application of nanoparticles of the ethanolic extract to improve bioavailability in the investigation of antiproliferative and pro-apoptosis effects in T47D breast cancer cell lines, as well as studies with essential oil in colorectal cancer cell lines (HT-29). It was found that OE can be a protector of DNA damage in 3T3-L1 fibroblast cells, depending on the applied concentration (Hasibuan & Sumaiyah, 2019;Thirugnanasampandan et al., 2015).

Diuretic e nephroprotective activities
The diuretic and nephroprotective activities of the ethanolic extract of the leaves were tested using furosemide as a standard drug in nephropathies induced acetaminophen. The research showed an increase in the total production of urine and urinary electrolytes, proving the nephroprotective and antioxidant effects (Palani et al., 2010).
The studies by Amarasiri et al. (2018), El-hawary et al. (2012 and Patel et al. (2010b) evaluating the nephroprotective and diuretic effects of aqueous and ethanolic leaves extracts also showed a significant increase in urinary volume and urinary electrolytes. Analyses with alcoholic leaves extract showed promising results in cisplatin-induced nephropathies, suggesting that the extract increases the expression of TGF-1β (transforming growth factor-1β) and, consequently, inhibits renal necrosis and cellular infiltration (Sahrial & Solfaine, 2019).
The extract was mixed with a solution of zinc nitrate and submitted to the drying process until obtaining the nanoparticles, which showed safe and satisfactory results against A. stephensi, C. quinquefasciatus and C. tritaeniorhynchus. In this proposal, a formulation was developed to achieve better selectivity and less side effects (Vijayakumar et al., 2015).
Regarding antileishmania activity, two articles described this action in the essential oil, in vitro and in vivo, against Leishmania (Viannia) braziliensis. The OE presented antileishmania activity in tests in vitro and in vivo. The oil inhibited the growth in vitro of 107 promastigotes of Leishmania braziliensis after 48 hours, by intralesional administration for 14 days. The ethyl acetate extract from leaves also caused an in vitro and in vivo reduction in the number of promastigotes, in a shorter treatment time, and can be an additive to the existing therapy (Gonçalves, 2017;Lima et al., 2014).
Alcoholic extracts from leaves, roots and stems showed good anthelmintic activity against Pheritima posthuma (intestinal parasite). The response was close to the control drugs used in the study, piperazine citrate and albendazole, with the most potent action attributed to the leaves extract and the least potent to the root extract (Prasenjit et al., 2011).

Other identified actions
The improvement of animal feed with the addition of indian borage leaves to obtain weight gain in domestic chickens, are applications for veterinary use. The lowest amount of total coliforms and CFU (colony-forming units) detected in the feces of these birds, was observed in the group with antibiotic-free diet, which were replaced by the leaves extract. Therefore, this study proves the bacteriostatic efficiency of the leaves extract, reducing the bacterial count in the feces and indicating a safe alternative to replace antibiotics in poultry feed (Chuchuca et al., 2016).
Another reported veterinary use is of leaves used in vitro as a modulator of rumen fermentation. The addition of leaves to the diet can reduce methane generation by up to 30% and increase the digestion of dry matter, ammonia and other fermentation substances, such as volatile fatty acids, whose isomers are beneficial for human health (Yanza et al., 2018).
The study of psoriasis reports an ointment formulation made from ethanolic extract of roots. The antipsoriasis effect was proven to induce psoriasis with topical application of a mixture containing formaldehyde for seven days on the dorsal skin of mice. The effect of ointments containing indian borage extract at concentrations of 0.5% and 1%, was evaluated by the severity index of psoriasis, its phenotypic characteristics such as redness, scales and erythema and the aspects of the epidermal layer of the skin in vivo. There was a progressive reduction in the severity of psoriasis lesions, showing good response to its treatment and confirming its traditional use in skin diseases (Vijayalakshmi et al., 2019).
The hepatoprotective effects of the ethanolic and aqueous extract from leaf were tested to induce hepatotoxicity in rats with carbon tetrachloride (CCl4). Animals with altered liver function in this research were monitored using biochemical markers (aspartate transaminase, alanine transaminase, alkaline phosphatase, gamma-glutamyl transpeptidase and total and direct bilirubin), and those treated with ethanol extract showed more effective recovery of these markers, in compared to those treated with the aqueous extract. And once the hepatocytes have regenerated and the fat changes have normalized, validating the popular use of this species due to its hepatoprotective effect (Patel, 2011).
The antidiabetic activity, in ethanol and acetone extracts of leaves, by alpha-amylase inhibition was confirmed in vitro (Bole & Kumudini, 2014). Therapeutic applications of this species can expand its use as a functional nutraceutical food (Bhatt et al., 2013).
This study synthesized the existing knowledge in scientific publications about P. amboinicus and its therapeutic properties, as well as identified the existing processes and products involving this species and its applications in order to stimulate innovations in biotechnology.
This prospecting revealed that the benefits of this species can be seen in both humans and animals, with different purposes. The scientific publications analyzed showed that some therapeutic actions of popular use, although tested and validated, are not yet described in patent documents. The good therapeutic performance of this plant contributes to future studies where the information provided here can be used for the developing of innovative products.

Conclusions
There was an interdependence between the information found in patent databases and academic research about P.
amboinicus and its therapeutic actions. And the association of technology with scientific research is essential to validate the traditional use of this species by the population.
These data can leverage research for the generation of pharmaceutical patents, be they process or product patent and the development of products with cosmetic or food applications. Therefore, studies are promising due to the many potentialities of this plant that have not yet been fully elucidated and explored. and the Federal University of Maranhão (UFMA) for the support and infrastructure provided for research, analysis and data discussion.