ES2389070B1 - DERIVED FROM PRENILATED DIBENZOFURANIC POLYCETIDES AND THEIR APPLICATION FOR THE TREATMENT OF INFECTIOUS, PARASITIC DISEASES AND TUMORS. - Google Patents

Abstract

Antimicrobial, antitumor and antiparasitic activity of derivatives of prenylated dibenzofuranic polyketides. # The present invention describes a family of compounds that are naturally occurring prenylated dibenzofuranic polyketides that possess antimicrobial activity, particularly against gram positive bacteria. Likewise, these compounds present cytotoxic activity against various tumor cell lines, as well as antiparasitic activity, for which reason they are useful for the manufacture of antitumor drugs and for the treatment of infectious diseases of bacterial and parasitic origin.

Description

Derivatives of preylated dibenzofuranic polyketides and their application

for the treatment of infectious and parasitic diseases and tumors.

The present invention describes a family of compounds that are polyketides

5

Prenylated dibenzofuranics of natural origin that possess activity

antimicrobial, particularly against gram positive bacteria, so they are

useful in the manufacture of drugs for the treatment of diseases

infectious of bacterial origin. Likewise, these compounds present

cytotoxic activity against various tumor cell lines, as well as

1O

antiparasitic activity.

STATE OF THE PRIOR ART

The progressive increase in bacterial infections caused by strains

fifteen

resistant to multiple antibiotics and by highly virulent strains it has been

become one of the most serious health problems in patients

hospitalized. The selective pressure resulting from the extensive use of antibiotics

over the past 25 years has led to the emergence of a multitude of

bacterial resistance and the spread of resistance genes among

twenty

pathogenic microorganisms. All this has caused us to meet

at risk of "intractable" infections. The growing appearance and

spread of resistance in Gram-positive bacteria such as genera

Enterococcus, Staphylococcus and Streptococcus are clear examples of

it. The constant and rapid worldwide dispersal of microbial strains

25

carriers of multiple resistance and virulence genes has caused a

large increase in morbidity and mortality caused by infections

hospitals, as the WHO already pointed out in 1999 in its quot; Report on the

Infectious diseases. Remove Obstacles to Healthy Developmentquot; and in

2001 in the document `` lnfectíon control programs to control antimícrobíal

30

resist ''.

Currently, one of the most dramatic examples of this situation is

constitutes the increasing emergence and spread of resistance to antibiotics and

of virulence factors in Staphylococcus aureus and other members of the

35

genus Staphylococcus. Staphylococci are ubiquitous microorganisms,

present in the respiratory tract of 30% of healthy adults and on the skin of

20%. Patients and hospital staff present percentages somewhat

Taller. However, multiple population groups are at high risk for pathogenic staphylococcal infections. The development of staphylococcal infections among newborns and their mothers, in transplant patients and in people affected by diseases such as influenza, leukemia, diabetes mellitus, bronchopulmonary pathologies, etc. is normal. Although within this bacterial genus, the species S. aureus is the cause of the greatest number of staphylococcal pathogenesis, various species of coagulase negative Staphylococcus (CoNS) have been associated with an increasing number of infections acquired in hospitals. The increasingly frequent use of invasive procedures, involving different prosthetic materials, and the high number of immunosuppressed patients have contributed to the dramatic increase in infections due to CoNS. For all that has been said, the control and treatment of infections caused by Staphylococcus constitutes a topic of extreme clinical relevance. In addition, already in the 21st century, the increasing spread of strains resistant to methicillin, an antibiotic that was the basic agent against such infections, has made their eradication extremely difficult. In both the USA and Japan and Europe, there is an increasing frequency of the appearance of isolates of methicillin-resistant S. aureus (MRSA), especially in the intensive care units of hospitals, although the appearance of infections is also increasing acquired in the community.

MRSAs are frequently resistant to ~ -lactam antibiotics, such as penicillins, cephalosporins, and carbapenenes. Resistance to aminoglycosides and macrolides is also common. Few antibiotics are still effective against MRSA. Specifically, mupirocin and glycopeptide antibiotics, mainly vancomycin, are the alternative treatment commonly used. However, the recent isolation in the USA of three strains of S. aureus with high resistance to vancomycin (HVRSA) acquired from Enterococcus, from multiple strains of S. aureus resistant to intermediate levels (IVRSA) of vancomycin in different countries, as well as the The increasing appearance and spread of staphylococci resistant to mupirocin (MuRSA) means that we are facing a situation of global risk for public health. Faced with this compromise, new antibiotics such as linezolid or daptomycin are being tested in the clinic, but a short time after their introduction, resistance to them appears. Likewise, in these bacteria the dispersion of virulence genes, such as genes encoding toxins, and resistance genes is increasingly common because many of these genes are encoded in mobile genetic elements, such as conjugated plasmids, islands pathogenicity, phages, etc.

The success of natural products over synthetic ones lies in the fact that they have been selected during evolution to interact with biological molecules, being considered as sources of privileged structures. Therefore, the study of plant species of interest due to their ethnobotanical use constitutes a valuable starting point in the search for products that can be converted into drugs. In this sense, traditional Argentine medicine uses many medicinal species to counteract various diseases. Achyrocline satureioides (Lam.) D. C, commonly known as "marcela" and a member of the Asteraceae family, it is a representative example within the widely used species.

In the last two decades, A.satureioides has been the subject of intense scientific research using both in vitro and in vivo animal models, providing experimental evidence that the extracts of this species have a wide spectrum of pharmacological and therapeutic properties. Among the properties registered so far, the following can be mentioned: hepatoprotective, activity associated with the antioxidant properties of this species, as well as choleretic, muscle relaxant, analgesic and sedative, antihyperglycemic, photoprotective, anti-inflammatory and immunomodulatory activity. There are data on insecticidal activity on Triatoma infestans and antiparasitic activity against the infectious form of Trypanosoma cruzí. Various works demonstrate anti-herpetic activity and against HIV-1, cytotoxic activity against human liver carcinoma cells, as well as antibacterial activity against Gram-positive and Gram-negative strains.

DESCRIPTION OF THE INVENTION

The present invention describes a group of compounds that are naturally occurring prenylated dibenzofuranic polyketides. These compounds have antimicrobial activity with high specificity against gram positive bacteria, including the multi-resistant strain Staphylococcus aureus. in addition

These compounds show cytotoxic activity against various cell lines from leukemia and breast cancer (HL-60; MCF7, SKBR3 and HEL, among others), as well as antiparasitic activity against Trypanosoma cruzí, the etiological agent of Chagas disease.

In a first aspect, the present invention relates to a compound of formula (1):

where R1, Rs, R6 and R7 are independently selected from hydrogen, O, OH,

Halogen, C1-C5 alkyl, C1-C6 alkenyl, aryl, O-C1-C12 alkyl, OC (O) R, N (R ') (Rquot;); where R is selected from C1-C5 alkyl, O-C1-C12 alkyl, aryl or heteroaryl and R 'and Rquot; independently selected from H or C1-C6 alkyl, R2 and R3 can be independent groups or be linked to form a ring;

When R2 and R3 are independent they are selected from hydrogen, OH or C1-C6 alkyl; when R2 and R3 are linked forming a ring they are selected from O or C (Rs) (Rg), Rs and Rg being independently H or C1-C6 alkyl; R4 is selected from H, C1-C6 alkyl or C1-C6 alkenyl;

20 -------- represents a bond that can be single or double;

or a pharmaceutically acceptable salt, solvate or stereoisomer thereof.

The term "alkyl" refers, in the present invention, to radicals of hydrocarbon chains, linear or branched, having from 1 to 12 carbon atoms, preferably from 1 to 6, and which are attached to the rest of the molecule

by single bond, eg methyl, ethyl, n-propyl, i-propyl, n-butyl, tert-butyl, sec-butyl, n-pentyl, n-hexyl etc. Alkyl groups can be optionally substituted by one or more substituents such as alkyl, aryl, halogen (called haloalkyl), hydroxyl, alkoxy, carboxyl, carbonyl, cyano, acyl, alkoxycarbonyl, amino, nitro, mercapto, and thioalkyl.

The term "arylo" refers, in the present invention, to single or multiple aromatic rings, having between 5 to 18 links in which a proton has been removed from the ring. Aryl groups are for example, but not limited to, phenyl, naphthyl, diphenyl, indenyl, phenanthryl, fluorenyl, or anthracyl. Preferably the aryl group has 5 to 7 carbon atoms and more preferably the aryl group is a phenyl. Aryl radicals can be optionally substituted by one or more substituents such as alkyl, halogen, hydroxyl, or carboxylic acid.

The term "heteroaryl" refers to an aryl that contains at least one atom other than carbon, such as S, N or O, forming part of the aromatic ring.

The term "alkenyl" refers to radicals of hydrocarbon chains of 1 to 12 carbon atoms, preferably 1 to 6, containing one or more carbon-carbon double bonds, for example vinyl, 1-propenyl, allyl, isoprenyl, 2-butenyl, 1 , 3-butadienyl etc. Alkenyl radicals can be optionally substituted by one or more substituents such as halo, hydroxyl, alkoxy, carboxyl, cyano, carbonyl, acyl, alkoxycarbonyl, amino, nitro, mercapto, and alkylthio.

By "halogen" In the present invention, a bromine, chlorine, iodine or fluorine atom is understood.

Preferably R1, Rs and R7 are OH.

Preferably R2 and R3 are linked in a ring. More preferably, R2 is a group O and R3 is a group C (Rs) (Rg), with Rs and Rg independently being H or methyl, and more preferably R4 is hydrogen.

Preferably R6 is a C1-C4 alkenyl. More preferably, R6 is -CH2-CH (OH) -C (CH3) = CH2

Preferably R2 and R3 are OH.

Preferably R4 is a C1-C4 alkenyl. More preferably, R4 is -

C (CH3) = CH2

Preferably, R6 is a C1-C4 alkenyl. More preferably, R6 is -CH2-CH = C (CH3) 2

Preferably, the compound of formula (1) is selected from: 10 • 2-methyl-1- [6,7,9-trihydroxy-8- (2-hydroxy-3-methyl-but-3-enyl) -3, 3-dimethyl-10- (2-methyl-butyryl) -3H-4, 11-dioxa-benzo [a] fluoren-5-yl] -butan-1-one.

• 2-methyl-1- [1, 3, 7,9-tetrahydroxy-4- (2-hydroxy-3-methyl-but-3-enyl) -8- (3-methyl-but -2-en i 1 ) -6- (2-methy1-butiri 1) -dibenzofu ran-2-i 1] -butan-1 -one.

The terms "pharmaceutically acceptable salt", "solvato" or "stereoisomer" refers to any pharmaceutical salt, solvate, stereoisomer, or any other compound that, being administered to a recipient, is capable of providing (directly or indirectly) a compound described herein. However it will be appreciated that pharmaceutically unacceptable salts are

20 also within the scope of the invention since the latter may be useful in the preparation of pharmaceutically acceptable salts. The preparation of salts, stereoisomers and derivatives can be carried out by means of methods known in the art.

For example, the pharmaceutically acceptable salts of the compounds provided herein are synthesized by conventional chemical methods from a parent compound that contains a basic or acid residue. These salts are generally prepared by reacting the free acid or base form of the compounds with an amount

30 stoichiometric of suitable base or acid in water or an organic solvent or a mixture of both. Nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are generally preferred. Examples of acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromohydrate, iodohydrate, sulfate, nitrate, phosphate, and salts.

Of organic acids such as acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate and p-toluenesulfonate. Examples of base addition salts include inorganic salts, such as sodium, potassium, calcium, ammonium, magnesium, aluminum, and lithium salts, and organic base salts, such as ethylenediamine, ethanolamine, N, N-dialkylenethanolamine, triethanolamine, glucamine and salts of basic amino acids.

The compounds of the invention may be present in crystalline form as free compounds or solvates and both forms are understood to be within the scope of the present invention. The solvation methods are of general knowledge in the field. Suitable solvates are those that are pharmaceutically acceptable. In a particular embodiment, a solvate is a hydrate.

The compounds of the present invention, represented by formula (1), described above can include enantiomers, depending on the presence of chiral centers in each C, or isomers, depending on the presence of multiple bonds (for example, Z, E) . Individual isomers, enantiomers or diastereomers and mixtures thereof are included within the scope of the present invention.

The present invention further provides pharmaceutical compositions comprising compounds of formula (1) or pharmaceutically acceptable salts, derivatives, prodrugs, solvates or stereoisomers thereof together with a pharmaceutically acceptable carrier, adjuvant or vehicle for administration to a patient. Preferably, said composition also comprises another active principle with a synergistic or complementary effect.

The pharmaceutical compositions can be administered by any appropriate route of administration, for example, orally, topically, rectally, or parenterally (including subcutaneous, intraperitoneal, intradermal, intramuscular, and intravenous).

Pharmaceutical forms suitable for oral administration include any solid composition (tablets, lozenges, capsules, granulated forms, etc.) or liquid (solutions, suspensions, emulsions, syrups, etc.) and may contain conventional excipients known in the art, such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers, for example lactose, sugar, cornstarch, calcium phosphate, sorbitol or glycine, lubricants for the preparation of tablets, for example magnesium stearate, disintegrants such as starch, polyvinylpyrrolidone, sodium starch glycolate or microcrystalline cellulose, or pharmaceutically acceptable wetting agents, such as sodium lauryl sulfate.

Solid oral compositions can be prepared by conventional methods of mixing, filling, or tableting. Repeated mixing operations can be used to evenly distribute the active ingredient using large amounts of fillers. These operations are conventional in the art of this invention. Tablets can be prepared, for example through wet or dry granulation, and can optionally be coated by methods well known in normal pharmaceutical practice, particularly with an enteric coating.

The pharmaceutical compositions can also be adapted for parenteral administration, such as sterile solutions, suspensions or lyophilized products in suitable dosage form. Suitable excipients, such as bulk agents, neutralizers or surfactants may be mentioned.

The compounds or compositions described in the present invention can be administered by any suitable method, such as intravenous infusion, oral preparations, and intraperitoneal or intravenous administration. However, the preferred route of administration will depend on the condition of the patient. In particular, oral administration is preferred due to the comfort for the patient and the chronic nature of the diseases to be treated.

For their therapeutic application, the compounds of formula (1) should preferably be in pharmaceutically acceptable or substantially pure form, for example the compounds of formula (1) have a pharmaceutically acceptable level of purity excluding the excipients admitted and not including material considered toxic to normal dosage levels. The purity levels of a compound of formula (1) or its derivatives, preferably exceeds 50%, even more than 70%, and even 90%. In a preferred representation, that exceeds 95%.

The therapeutically effective amount of the compound of formula (1) to be administered in general will depend, among other factors, on the person to be treated, the severity of the disease, the chosen form of administration, etc. For this reason, the doses mentioned in this invention must be considered as guides for the specialist in the field and the latter must adjust the dose according to the variables mentioned above. However, a compound of formula (1) can be administered one or more times a day, for example, 1, 2, 3 or 4 times a day in a typical total daily amount between 1 and 200 mg / kg of body weight. / day, preferably 1–10 mg / kg body mass / day. In the same way a compound of formula

(11) can be administered one or more times a day, for example, 1, 2, 3 or 4 times a day in a typical total daily amount between 1 and 200 mg / kg of body weight / day, preferably 1-1 Or mg / kg of body mass / day.

The compounds described in this invention, their pharmaceutically acceptable salts, stereoisomers and / or solvates, as well as the pharmaceutical compositions containing them can be used in conjunction with other additional drugs to provide combination therapy. Said additional drugs can form part of the same pharmaceutical composition or, alternatively, be provided in the form of a separate composition for their simultaneous administration or not, with the pharmaceutical composition comprising a compound of formula (1) or a pharmaceutically acceptable stereoisomer, solvate. or get out of it.

Another aspect of the invention is the use of a compound of formula (1) for the manufacture of a medicament.

Another aspect of the invention is the use of a compound of formula (1) for the manufacture of a medicament for the treatment of an infection of bacterial origin. Preferably, if the bacteria causing the infection is gram positive. Preferably, if the bacterium is selected from the genera Staphylococcus, Enterococcus and Streptococcus. Even more preferably, the bacterium is selected from Staphylococcus aureus and Enterococcus faecalís. Also within the scope of the invention are bacteria of these genera that have developed resistance to certain antibiotics such as, but not limited to, vancomycin and methicillin.

Another aspect of the invention is the use of a compound of formula (1) for the manufacture of a medicament for the treatment of cancer. Preferably, the cancer is leukemia or breast cancer.

A final object of the invention consists of compounds of formula (1) or pharmaceutically acceptable salts, derivatives, solvates or stereoisomers thereof for use as a medicine in patients with parasitic diseases and fungal diseases, preferably diseases caused by the Trypanosoma cruz parasite. .

Throughout the present description, the term "treatment" refers to eliminating, reducing or diminishing the cause or effects of a disease. For the purposes of this invention, treatment includes, but is not limited to, alleviating, diminishing, or eliminating one or more symptoms of the disease; reduce disease grade, stabilize (i.e. not worsen) disease state, delay or slow disease progression, alleviate or improve disease state, and remit (either total

or partial).

Throughout the description and claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will emerge in part from the description and in part from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.

DESCRIPTION OF THE FIGURES

Fig. 1. Shows the sensitivity patterns of compound C2 of this invention against different gram-positive clinical strains and multiple S. aureus strains that differ in their antibiotic resistance profiles (table under the graph).

Fig. 2. Shows the growth levels of the standard strains of S. aureus and

Methicillin-resistant and partially vancomycin-resistant E. faecalís and S. aureus (IVRSA); versus a logarithmic plot of the different concentrations of compound C2.

Fig. 3. Shows growth levels against a logarithmic plot of the different concentrations of ampicillin (control).

Fig. 4. Shows the growth levels against a logarithmic plot of the different concentrations of kanamycin (control).

EXAMPLES

Obtaining the compounds of formula (1)

Metabolites with the general structure (1) were isolated from leaves and stems of Achyroclíne satureíoídes (Lam.) OC. The structure of the metabolites has been numbered according to the methodology commonly used in the characterization of natural products, using common names proposed in the bibliography (Giovanni Appendino et al. Quot; Arzanol, an Anti-inflammatory and Anti-HIV-1 Phloroglucinol a-Pyrone from Helíchrysum ítalícum ssp. Mícrophyllumquot ;, J. Nat. Prod. 2007, 70, 608-612 -John R. Carney et al. Quot; Achyrofuran, a New Antihyperglycemic Dibenzofuran from the South American Medicinal Plant Achyroclíne satureídesquot ;, J. Nat. Prod. 2002, 65, 203-205).

Example 1 Compound C1

Cl

Viscous yellowish oil. 1H-NMR (lgt ;, CDCb): 15.9 (1H; s; OH-9); 14.45 (1H; s; OH-3); 10.07 (1H; s; OH-1); 6.70 (1H; d; J = .9.8 Hz; H-1 "'); 5.65 (1H, d, J = 9.8 Hz, H-2 "'); 4.98 (1H; s; H-

5 4a ') 4 80 (1 H · s · H-4b') · 4 44 (1 H · sa · H-2 ') · 3 95 (1 H · sext · J = 6 5 Hz · H-2quot ; quot;)

'' '' '' '' '' '' '' '3.85 (1H; sext; J = 6.7 Hz; H-2' '); 3.15 (2H; dd; J = 14.2 Hz; H-1 '); 1.92 (2H, m;

H-3bquot; H-3b "") · 1 89 (3H · s · H-5 ') · 1 59 (3H · s · H-4``) · 1 58 (3H · s · H-5``) · 1 quot; 47

'' '' '' '' '' '' '' '(2H · m · H-3aquot; H-3aquot; quot;) · 1 25 (3H · m · H-5``) · 1 23 (3H · mH-5 "quot;) · O97 (3H · m · H-

, '' '' '' '' '' '' '' 4 '' quot;); 0.96 (3H; m; H-4 "). 13C-NMR (lgt;, CDCI3): 212.1 (s, C-1 quot;); 212.6 (s, C-1 ""); 10 163.3 (s, C-3); 159.4 (s, C-1); 155.8 (s, C-5a); 156.1 (s, C-9); 154.2 (s, C-7); 152.3 (s, C-4a); 147.2 (s, C-3 '); 127.1 (d, C-2'quot;); 115.5 (d, C-1 "); 110.6 (t, C-4 '); 106.7 (s, C-9a); 106.5 (s, C-2, C-8); 103.3 (s, C-9b); 102.3 (s, C-4); 100.4 (s, C-6); 79.2 (s, C-3 "'); 75.4 (d, C-2 '); 46.4 (d, C-2 "quot;); 46.2 (d, C-2 "); 29.9 (t, C-1 '); 27 8 (e C-4 "C-5") · 26 9 (t C-3 "") · 26 7 (t C-3 ") · 18 1 (e C-5 ') · 16 6 (e C-5``)

'' '' '' '' '' '' '' '' 15 16.6 (e, C-5 "quot;); 11.9 (e, C-4 "); 11.9 (e, C-4 "quot;). HREIMS: 550.2607 (calcd for C32H3sÜs [M] +, 550.2567). IR Vmax: 2964, 2932, 2879, 1772, 1724, 1620, 1459, 1409, 1373, 1176, 1105, 1038, 950 cm -1. UV (EtOH) Amax (log E): 341 (2.53);

239 (2.55); 203 (2.65) nm. [a] 0 = -2.3 (e 0.008; CHCl3).

Example 2

Compound C2

C2

Viscous yellowish oil. 1H-NMR (5, CDCb): 15.63 (1H; bs; OH-7); 13.98 (1H; s; OH-3); 9.82 (2H, bs; OH-1, OH-9); 5.30 (1H; m; H-2 '); 5.09 (1H; s; H-4a'quot;); 4.96 (1H; s; H-4b'quot;); 4.53

5 (1H 2``) 3 44

'' '' '' '' '' '' '' '' (2H; d; J = 6.3; H-1 '); 3.25 (1H; dd; J = 15.2; H-1 a'quot;); 3.04 (1H, dd, J = 15.2, H-

1 b'quot;) 1 90-1 87 (2H · m · H-3b "H-3b" quot;) 1 88 (3Hs -5 ') 1 70

'' '' '' '' '' '' '' '' (3H · s · H-4 ') · 1 54 (1 H · m · H-3aquot;) · 1 48 (1 H · m · H -3a "quot;) 1 26 (3H · d · J = 6 2 · H-

, '' '' '' '' '' '' '' '5 ") · 1 23 (3H · d · J = 6 8 · H-5" quot;) · O95 (3H · m · H-4 " ") · O93 (3H · m · H-4") 13C-

, '' '' '' '' '' '' '. 10 NMR (5, CDCb): 213.4 (s, C-1 ""); 207.1 (s, C-1 quot;); 163.9 (s, C-3); 157.6 (s, C-7); 155.5 (s, C-9); 158.2 (s, C-5a); 154.6 (s, C-1); 154.2 (s, C-4a); 145.8 (s, C-3'``); 132.1 (s, C-3 '); 122.2 (d, C-2 '); 111.7 (s, C-2); 111.7 (t, C-4'quot;); 107.4 (s, C-8); 105.2 (s, C-9b); 104.7 (s, C-9a); 102.3 (s, C-6); 101.3 (s, C-4); 78.1 (d, C-2'``); 46.3 (d, C-2 "quot;); 45.4 (d, C-2 "); 30.3 (t, C-1 "'); 27.1 (t, C-3 ""); 26.4 (t, C-3 "); 25.8 15 (e, C-4 '); 21.8 (t, C-1 '); 18.2 (e, C-5'quot;); 17.9 (e, C-5 '); 17.0 (e, C-5 "); 16.3 (e, C-5 "quot;); 11.8 (e, C-4 ", C-4" "). EIMS m / z (%): 552 (M +, 17); 534 (11); 479 (31); 425

(1 00); 389 (11). HREIMS: 552.2740 (calcd for C32H4OUs [M] +, 552.2723). IR Vmax: 3296, 2968, 2929, 2877, 1727, 1617, 1451, 1373, 1236, 1192, 1086, 1 20

1041, 905, 849 cm-. UV (EtOH) Amax (log t): 341 (2.69); 243 (2.67) nm. [a] 0-20 = +31.4 (e 0.116; CHCl3).

Biological tests

Antimicrobial activity assay

For the determination of MIC50, the microbial growth in microplates of

5 96 wells was monitored in the presence of between 6 and 10 different concentrations (range between 1 and 128 micromolar) of the tested compounds; together with the commercial antibiotics ampicillin and kanamycin as controls. For this, fresh cultures of bacteria (or yeasts) were diluted to optical densities at 620 nanometers of 1 or -5 per milliliter (equivalent to 105

10 colony forming units per milliliter); at which time they are cultured again in the presence of the tested compounds, or the control antibiotics. Growth readings were taken after 24 hours. Then, the growth values (expressed in optical density at 620 nanometers) are plotted against the logarithm in base

15 10 of the concentrations of the compounds tested. The points of the graph thus obtained are adjusted to the inverse sigmoid curve to obtain the definitive MIC50 value. These tests were carried out with two gram-negative bacteria (Escheríchía colí and Pseudomonas aerugínosa), two gram-positive bacteria (S. aureus and Enterococcus faecalís), and the yeast Saccharomyces

20 cerevísíae as a eukaryotic (not bacterial) model. The strain of E. coli used carries a plasmid that confers resistance to ampicillin. Table 1 shows the MIC50 values of the compounds mentioned in this invention.

Table 1: MIC of compounds of the invention tested (~ M)

Compound

E. coli P. aerugínosa S. aureus E. faecalís S. cerevísíae

Compound

C1 >> 100 >> 100 9 ± 1 48 ± 18 > 100

C2

>> 100 >> 100 <1 <1 > 100

Ampicillin

>> 100 89 ± 55 <1 <1 NO

Controls

Kanamycin <1 114 ± 20 <1 23 ± 4 NO

The sensitivity of different clinical strains of gram-positive bacteria against the compounds of this invention was studied. The tests were carried out as mentioned for MIC5o, but only three concentrations were included. In this case, the relative growth was calculated for each concentration in comparison with control cultures, without compounds.

added. Figure 1 shows the sensitivity patterns of different Gram-positive clinical strains; and multiple strains of S. aureus, which differ in their antibiotic resistance profiles, against compound C2 described in this invention. Table 2 details the antibiotic resistance profile of each

5 strain tested.

For the growth inhibition assays (MIC50) of compound C2 of this invention against a strain of S. aureus resistant to methicillin, and partially resistant to vancomycin (IVRSA); the inhibitor was added in a range of 24 different concentrations ranging from 10 nanomolar to 100 micromolar. The commercial antibiotics ampicillin and kanamycin were used as controls in the same concentration range. Standard strains of S. aureus and E. faecalís were included in the trial. Figure 2 shows the growth levels against a

15 logarithmic plot of the different concentrations of compound 2. Figure 3 shows the growth levels versus a logarithmic plot of the different concentrations of ampicillin (control). Figure 4 shows the growth levels against a logarithmic plot of the different concentrations of kanamycin (control).

Cell viability assay by MTT.

Different cancer cell lines (HL60, MCF7, SK-Br3, HEL and others) were cultured in RPMI and DMEM medium containing 10% fetal bovine serum. For MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-5 diphenyltetrazole bromide) analysis (Sigma-Aidrich, St. Louis, MO), cells were seeded in 96-well plates at a concentration of 10000 cells / well. After 24 h of incubation at 37 oC and 5% C02, the solvent (DMSO, at a final non-toxic concentration O, 1%) or the predetermined range of concentrations of the compound to be tested was added. After 48 h of incubation,

10 added a MTT solution to each well (final concentration 0.5 mg / ml), and the plates were further incubated 3 h at 37 oC. Subsequently, the medium was extracted and the formazan crystals were solubilized by adding SDS-HCI (20% SOS; 0.02M HCI) to each well. The absorbance of each well was recorded by means of a reader of

15 microplates at 570 nm. For the calculation of IC50 (Inhibitory Concentration 50), a non-linear regression analysis was performed using the GraphPad v5 program.

Thus, by way of example, Table 3 shows cytotoxicity data exerted by compound C2.

Table 3: IC50 of compound C2 described in the present invention (~ M)

Compound

Cell Lines

HL60

MCF7 SKBr3 HEL

C2

1.4 > 30 2.5 1.1

Claims (24)

1. Compound of formula (1): or

(1)

where

5

R1, Rs, R6 and R7 are independently selected from hydrogen, O,

OH, halogen, C1-C5 alkyl, C1-C6 alkenyl, aryl, O-C1-C12 alkyl,

OC (O) R, N (R ') (Rquot;); where R is selected from C1-C5 alkyl, O-alkyl

C1-C12, aryl or heteroaryl and R 'and Rquot; are independently selected

between H or C1-C5 alkyl,

10

R2 and R3 can be independent groups or be linked to form a

cycle;

when R2 and R3 are independent they are selected from hydrogen, OH

or C1-C6 alkyl;

when R2 and R3 are joined to form a cycle, they are selected from O or

fifteen

C (Rs) (Rg), where Rs and Rg are independently H or C1-C6 alkyl;

R4 is selected from H, C1-C6 alkyl or C1-C6 alkenyl;

-------- represents a bond that can be single or double,

or a pharmaceutically acceptable salt, solvate or stereoisomer of the

same.

twenty

2.

Compound according to claim 1 where R1, Rs and R7 are OH.

3.

Compound according to any of claims 1 or 2 where R2 and R3

they are united in a cycle.

Four.

Compound according to claim 3 where R2 is a group O and R3 is a group C (R8) (R9), R8 and R9 being independently H or methyl.

5.

Compound according to claim 4, wherein R4 is hydrogen.

6.

Compound according to any one of claims 1 to 5 wherein R6 is a C1-C4 alkenyl.

7.

Compound according to claim 6 where R6 is -CH2-CH (OH) -C (CH3) = CH2.

8.

Compound according to any of claims 1 or 2 where R2 and R3 are OH.

9.

Compound according to claim 8 where R4 is a C1-C4 alkenyl.

1O. Compound according to claim 9 where R4 is -C (CH3) = CH2.

eleven.

Compound according to any one of claims 8 to 1 Odonde R6 is a C1-C4 alkenyl.

12.

Compound according to claim 11 where R6 is -CH2-CH = C (CH3) 2.

13.

Compound according to claim 1 which is selected from: 2-methyl-1- [6,7,9-trihydroxy-8- (2-hydroxy-3-methyl-but-3-enyl) -3,3-dimethyl-1 0- (2-methyl-butyryl) -3H-4, 11-dioxa-benzo [a] fluoren-5-yl] -butan-1-one. 2-methyl-1- [1,3,7,9-tetrahydroxy-4- (2-hydroxy-3-methyl-but-3-enyl) -8- (3-methy1-but-2-en i 1) -6- (2-methy 1-butiri 1) -dibenzofu ran-2-i 1] -butan-1 -one.

14.

Pharmaceutical composition comprising a compound of formula (1) according to any of claims 1 to 13 together with a pharmaceutically acceptable carrier, adjuvant or vehicle.

fifteen.

Composition according to claim 14, which also comprises another active principle.

16.

Use of a compound of formula (1) according to any of claims 1 to 13 for the manufacture of a medicament.

17.

Use of a compound of formula (1) according to any of claims 1 to 13 for the manufacture of a medicament for the treatment of an infection of bacterial origin.

18.

Use according to claim 17 where the bacterium causing the infection is gram positive.

19.

Use according to claim 17 where the bacterium is selected from the genera Staphylococcus, Enterococcus and Streptococcus.

twenty.

Use according to claim 19 where the bacterium is selected from Staphylococcus aureus and Enterococcus faecalís.

twenty-one.

Use according to claim 20 where the bacterium is vancomycin resistant S. aureus.

22.

Compound according to any one of claims 1 to 13 for use as a medicine.

2. 3.

Use of a compound of formula (1) according to any of claims 1 to 13 for the manufacture of a medicament for the treatment of cancer.

24.

Use according to claim 23 where the cancer is selected from breast cancer or leukemia.

25.

Use of a compound of formula (1) according to any of claims 1 to 13 for the manufacture of a medicament for the treatment of a parasitic disease.

Table 2: Antibiotic resistance profiles of the clinical strains tested against compound 2