Antitumor Activity of Leaves fromHyptis mutabilis (A. Rich.) Briq. (Lamiaceae) inMice Bearing Tumor

eHyptis genus has more than 400 species, many of them being used in folk medicine to treat several conditions. Some anticancer compounds have been isolated fromplants of this genus, and for that reasonwe decided to investigate the potential in vivo antitumor activity of extracts of leaves of Hyptis mutabilis with different polarities (hexane, methanol, water, and hot water) against two mice tumors: sarcoma 180 and Ehrlich solid tumor. Phytochemical analysis revealed strong presence of steroids, saponins, �avonoids (mainly dihydro�avanols), and catechins. Acute toxicity was perfomed according to the up-and-downmethod showing LD50 values ranging from 100 up to 2500mg/kg. Antitumor activity was investigated using 10% of the LD50 for each extract. Methotrexate was used as positive control. Both aqueous extracts showed strong inhibition of tumor growthwith values up to 70% of inhibition growth for sarcoma 180. Ehrlich solid tumor was only slight inhibited by hexane extract (38.6%). In conclusion, the aqueous extracts ofH. mutabilis showed promising results against sarcoma 180 mice tumor.

From the phytochemical point of view, the genus has been extensively studied: terpenoids, including monoterpenes and sesquiterpenes present in essential oil, �avonoids, lactones, lignans, and alkaloids were isolated and identi�ed [12].e anticancer agents 3 � -demethyldesoxypodophyllotoxin and betulinic acid were isolated from plants of this genus [13].Based on these �ndings, we evaluated the antitumor activity of Hyptis mutabilis (A.Rich.)Briq.against murine tumors in these work.

Methodology
Male Swiss mice (20-25 g) were obtained from the animal facilities of the Department of Antibiotics of the Federal University of Pernambuco.All animals were kept under standard conditions, with temperature at 25 ± 2 ∘ C, 12 h light/dark cycle, and relative humidity, with food and water ad libitum.Experiments were carried out in accordance with the ethical principles of e Brazilian College of Animal Experiments and the National Institute of Health Guide for Care and Use of Laboratory Animals.is study was previously approved by the Committee of Experimental Animals of the Federal University of Pernambuco (004788/2005-68).
Plant material and extract were as follows.Leaves from H. mutabilis were collected in Aldeia (Camaragibe-PE).Botanical identi�cation was made by Professor Dr. Alda Andrade Chiappeta of the Department of Antibiotics, Federal University of Pernambuco.Voucher specimen was deposited at the Herbarium of the Department of Antibiotics (DAUFPE 6256).Leaves were dried in open air at room temperature (28 ± 2 ∘ C) and crushed with an electric grinder.Powdered leaves (940 g) were subjected to serial extractions with increasing polarity solvents: hexane, methanol, water, and hot water.e sample was extracted three times with each solvent (3 hours each under stirring), �ltered, and subjected to the next extraction.Hexane and methanol extracts were evaporated under low pressure at constant temperature of 45 ∘ C, yielding 17.64 g (1.88%) and 61.42 g (6.53%), respectively.Water extracts were liophilized yielding 44.01 g (4.68%) for water and 22.00 g (2.34%) for hot water.
For the phytochemical analysis we performed phytochemical screening according to Matos [14] and Sena Filho et al. [15], based on chemical tests, to detect the presence of common secondary metabolite classes, mainly alkaloids, terpenes, and phenolic compounds, in hexane and methanolic extracts, using classical speci�c colour reactions.
Acute toxicity was evaluated according to the up-anddown method to determine LD 50 values using fewer animals as possible (described in OECD 425).
Antitumor activity against Ehrlich solid tumor and sarcoma 180 was investigated.Ehrlich ascite carcinoma and sarcoma 180 were maintained in vivo in mice by transplantation of 0.2 mL of ascitic �uid (1 × 10 7 cells) from the infected mice to the noninfected mice [16].e Ehrlich ascites or sarcoma 180 ascites (S180) tumor cells (1 × 10 7 cells/mouse) were subcutaneously implanted into the inguinal region of the mice as described by Sugiura and Stock [17].Mice were divided into the respective groups and the treatment started aer 24 hours of tumor cells' implantation.
For the treatment of each tumor group, animals (  ) were divided in six groups: negative control (received 1% DMSO dissolved in saline solution 0.9%, i.p.), positive control (received 10 mg/kg of methotrexate, i.p.), and four groups treated with H. mutabilis extracts (10% of LD 50 found for each extract).Hexane extract group were treated with 250 mg/kg (i.p.), methanol extract group received 230 mg/kg (i.p.), and water and hot water extracts received 150 mg/kg and 10 mg/kg, respectively, both intraperitoneally to avoid food-extract interactions.Animals were treated once a day during seven days.On the eighth day, mice were weighed and killed and the tumors were removed.e inhibition of tumor growth was calculated by percentage of reduction of the tumors when compared with the negative control group.

Dataset Description
e dataset associated with this Dataset Paper consists of 3 items which are described as follows.).Phytochemical screening of methanolic and hexanic extracts of H. mutabilis leaves revealed the presence mainly of steroids, saponins, �avonoids (mainly dihydro�avonols), and catechins.In the table, "+" means weak; "++", medium; "+++", strong; and "0", absent.ese results are similar to those found for other Hyptis species [12].).LD 50 values obtained for H. mutabilis extracts according to OECD 425 protocol.Single-dose acute toxicity tested showed values of LD 50 ranging from 100 mg/kg to 2500 mg/kg.e dose used to test the antitumor activity of the extracts was 10% of the LD 50 ; at these doses, no histological changes were observed in mice treated for seven days.So, these doses were considered safe for use in the following in vivo experiments [18].

Dataset Item 1 (Table
Dataset Item 3 (Table ).In vivo growth inhibition percentual of sarcoma 180 (S180) and Ehrlich solid tumor (EST) of H. mutabilis extracts.Signi�cant reduction of sarcoma 180 tumor mass was obtained with water extracts (72.4 and 74.3% of inhibition of tumor growth for water and hot water extracts, resp.)aer 7 days of treatment.Methotrexate was used as standard drug, used as positive control, and completely inhibited sarcoma 180 tumor growth.Hexane and methanol extracts just slightly inhibit sarcoma 180 growth.Ehrlich solid tumor was only inhibited by hexane extract (38.6% of inhibition), while methotrexate inhibited 98.2% of tumor growth.Data were expressed as mean ± S.E.M.No difference in the weight of the animals was observed among the tested groups.ese results were obtained aer 7 days of treatment, during which the animals were monitored for the appearance of adverse reactions, common to major commercial anticancer drugs.In the mice treated with H. mutabilis extracts, no strong adverse effects were observed, while in methotrexate groups, animals presented behavioral changes and enterocolitis.In the table, "-" indicates no inhibition.

Concluding Remarks
Plants are rich source of antitumor agents and many of the antitumor drugs in the market are derived from them [19][20][21].Both water and hot water extracts of H. mutabilis showed promising antitumor activity against sarcoma 180 murine tumor.ese data point to the presence of polar compounds present in these extracts that are probably responsible for this activity.As no adverse effect or any histopathological changes were observed during the 7 days of treatment, these data will serve to put forward other research groups to make a phytochemistry approach to isolate polar compounds in others to characterize the polar active compounds present