BETAXANTHINES DERIVED FROM TRYPTOPHAN AND PHENYLETHYLAMINE FOR USE IN THE TREATMENT AND/OR PREVENTIO

专利摘要:
Betaxanthines derived from tryptophan and phenylethylamine for use in the treatment and/or prevention of cancer. Compounds of formula (I) are described, {IMAGE-01} of the family of betaxanthines, or of pharmaceutical compositions comprising them, for use in the treatment and/or prevention of cancer and/or tumors. Likewise, the use of nutraceutical compositions for the prevention of cancer or as chemopreventive compositions, comprising said compounds of formula (I), is described. (Machine-translation by Google Translate, not legally binding)
公开号:ES2802814A1
申请号:ES201930649
申请日:2019-07-12
公开日:2021-01-21
发明作者:Carmona Francisco García；Herrero Fernando Gandía；Escudero Paula Henarejos；García Samanta Hernández；Rubio María Alejandra Guerrero
申请人:Universidad de Murcia；
IPC主号:

专利说明:

[0001] BETAXANTHINES DERIVED FROM TRYPTOPHAN AND PHENYLETHYLAMINE FOR USE IN THE TREATMENT AND / OR PREVENTION OF CANCER
[0003] FIELD OF THE INVENTION
[0005] The present invention describes compounds of formula (I), of the family of betaxanthines, specifically betaxanthines derived from tryptophan and phenylethylamine, or of pharmaceutical compositions comprising them, for use in the treatment and / or prevention of cancer and / or tumors. . Likewise, the use of nutraceutical and food compositions for the prevention of cancer or as chemopreventive compositions, comprising said compounds of formula (I), is described.
[0007] BACKGROUND OF THE INVENTION
[0009] Cancer is a common and sometimes fatal disease. As the human population ages, the incidence of cancer increases, and with it the urgency to develop more potent and less toxic antineoplastic drugs. Plants are a source of phytochemicals which could be safer and more effective molecules in chemoprevention and treatment to improve the health of cancer patients. Certain phytochemicals are capable of inducing biological responses in animal cells, including enzyme inhibition, antioxidant activity, and regulation of cell signaling pathways (Pratheeshkumar et al., BioMed Research International, 2015. DOI: 10.1155 / 2015/324021) .
[0011] Betalains are water-soluble nitrogenous plant pigments that are characteristic of plants of the order Caryophyllales. This type of compound is produced when bethalamic acid - the structural unit of all betalains - fuses with amines or amino acids (Gandía-Herrero & García-Carmona, Trends in Plant Science, 2013. DOI: 10.1016 / j.tplants.2013.01.003 ).
[0013] Generally, betalains are classified into two groups depending on their color: betacyanins are all those betalains that have a violet color and betaxanthins are those that are yellow. However, the color little or nothing informs about the chemical structure of the molecule and its functional groups, being these (structure and groups) the authentic ones responsible for its chemical capacity and its biological activity.
[0014] From the structural point of view, betalains are classified into two large groups: (1) those that have a net positive charge because they comprise a quaternary ammonium group (iminium) in their structure, such as betacyanins or indicaxanthin (see Figure 1) ; and (2) those structures that have an imine with no positive net charge, among which are vulgaxanthins (I, II, III, IV), tryptophan-betaxanthine, phenylethylamine-betaxanthine, phenylalanine-betaxanthine and dopaxanthin (represented in Figure 2). Among the betaxanthines, a small group can be distinguished that are made up of a primary amine linked by an alkyl chain to an aromatic ring, such as tryptophanobetaxanthine, phenylethylamine-betaxanthine, phenylalanine-betaxanthine or dopaxanthin, structurally very different from those betaxanthins made up of a linear amine such as vulgaxanthins (I-IV) or humilixanthin (represented in Figure 2). These aromatic ring betaxanthines account for 7 molecules out of the total 31 identified in nature (cf. Table 1 of Gandía-Herrero & García-Carmona, Trends in Plant Science, 2013. DOI: 10.1016 / j.tplants.2013.01.003). On the other hand, the betacyanins represented in Figure 1 comprise Ra and Rb groups that can be complex structures of sugar and / or acid groups. These different structural characteristics can cause their behavior to differ greatly between them, even though they are classified in the same color group.
[0016] The state of the art describes various in vitro studies showing the activity of extracts containing betalains as antioxidants. Antioxidants act as electron donors that neutralize reactive oxygen species and other free radicals that could otherwise damage DNA and lead to oncogenesis. Many studies, both in cells and in preclinical trials, support the idea that antioxidant molecules are capable of protecting cells against carcinogenesis (Borek, Integrative cancer therapies, 2004. DOI: 10.1177 / 1534735404270578; ES2349522, 2011, García-Carmona et al .; Gandía-Herrero & García-Carmona, Trends in plant science, 2013. DOI: 10.1016 / j.tplants.2013.01.003). However, other studies link the proliferation of certain types of cancer to antioxidants, since the factor p53 causes an increase in reactive oxygen species, which triggers apoptosis, therefore, the disappearance of these reactive species due to antioxidants prevents apoptosis and causes cell proliferation (Valko et al., Chemico-biological interactions, 2006. DOI 10.1016 / j.cbi.2005.12.009; Sayin et al., Science translational medicine, 2014. DOI: 10.1126 / scitranslmed.3007653 ; Perera & Bardeesy, Nature, 2011. DOI: 10.1038 / 475043a). These disparate results indicate that unique studies should be carried out for each molecule classified as antioxidant, since it cannot be generalized that a molecule, because it has antioxidant effects in vitro or even inferred because it is structurally similar to another, is effective as an antitumor. The relationship between antioxidant capacity and antitumor activity is not direct. The antitumor activity is more complex and may be due to biochemical factors and various structural factors of each molecule tested, not related to its antioxidant power.
[0017] On the other hand, in the state of the art, plant extracts containing betalains, without purification, mainly betacyanins, such as betanin or betanidine, have been described (ES2320380; US2003036565; US6312697B1) for the treatment of cancer, since some betacyanins have been shown to have antioxidant, anti-inflammatory, hepatoprotective, anticancer, antidiabetic, antilipidemic, antimicrobial, radioprotective, and anti-proliferative properties (Gandía-Herrero, Escribano & García-Carmona, Critical Reviews in Food Science, 2016. DOI: 10.1080 / 10408398.2012.740103).
[0019] In particular, several publications show the anticancer and chemopreventive activity of various extracts containing betalains (Kapadia et al., Cancer letters, 1996. DOI: 10.1016 / 0304-3835 (65) 04087-0; Kapadia et al., Anti-Cancer Agents in Medicinal Chemistry, 2011. DOI: 10.2174 / 187152011795347504; SreeKanth et al., International Journal of Phytochemistry, 2007. DOI: 10.1016 / j.phymed.2007.03.017; Kapadia et al., Pharmacological Research, 2003. DOI: 10.1016 / S1043-6618 (02) 00285-2, Lechner et al., Journal of Medical Food, 2010. DOI: 10.1089 / jmf.2008.0280 and Zou et al., Nutrition Journal, 2005. DOI: 10.1186 / 1475-2891-4 -25). In addition, Opuntia fruit extracts (whose main pigment is indicaxanthin) have also shown their potential in the protection and recovery of the liver after damage has been induced in said organ (Galati et al., Phytotherapy Research, 2005. DOI: 10.1002 / ptr.1741). In general, said plant extracts contain, for the most part, betacyanins and the nature of the bioactive molecule (s) present in said extracts has not been reliably clarified.
[0020] However, some studies have been described in cancer cell lines using betalains partially purified from plant extracts. On the one hand, Farabegoli et al., (Farabegoli et al., Food chemistry, 2017. DOI: 10.1016 / j.foodchem.2016.09.112) partially purifies a beet extract giving rise to two fractions rich in betalains called fractions R1 and R2. In fraction R1 Farabegoli et al. identified vulgaxanthin I (glutamine-betaxanthin) and in the R2 fraction, betanin, isobetanin and betanidine. No other betalain is mentioned by the authors. Furthermore, Farabegoli et al., Do not show antioxidant or anticancer activity with individual components, but with the set of betalains and other molecules present, respectively, in the R1 and R2 fractions. Likewise, in the article by Khan and co-authors (Khan et al , LWT-Food Science and Technology, 2012. DOI: 10.1016 / j.lwt.2012.01.025) , two fractions rich in Rivina humilis are obtained from betalains, including proline-betaxanthine (indicaxanthin), 3,4-dihydroxyphenylalanine-betaxanthine (dopaxanthin), glutamic acid-betaxanthine (vulgaxanthin II), glutamine-betaxanthin (vulgaxanthin I), asparticobetaxanthin-5-hydroxanthine acid , betanin, betanidine, tyrosine betaxanthine and dopamine-betaxanthine (cf Khan et al., Table 1, p. 318). These authors describe the cytotoxic action of the extract of Rivina humilis berries in cancer cell lines, but not that of individual components, pigments or not.
[0022] Tryptophan-betaxanthine (Figure 2) is a betaxanthine with no net positive charge that has an aromatic ring, where betahalamic acid is condensed with the amino acid tryptophan (Gandía-Herrero & García-Carmona, Trends in Plant Science, 2013. DOI: 10.1016 /j.tplants.2013.01.003). This compound is present at the trace level in the Celosia argéntea plant (Schliemann et al., Phytochemistry, 2001. DOI: 10.1016 / S0031-9422 (01) 00141-8) and is found in the plants of traditional Chinese medicine (TCM ).
[0023] Other betaxanthines, such as phenylethylamine-betaxanthine and phenylalanine-betaxanthine (Figure 2), are present in very low amounts in some varieties of Opuntia ficus-indica fruits , although indicaxanthin (Figure 1) is the main pigment (Gandía-Herrero & García-Carmona, Trends in Plant Science, 2013, DOI: 10.1016 / j.tplants.2013.01.003). On the other hand, an in vitro antioxidant measure of tryptophan-betaxanthin has been described as a pure molecule, from which no bioactive or antitumor activity is inferred in vivo (Cai, Sun & Corke, Journal of Agricultural and Food Chemistry, 2003. DOI: 10.1021 / jf030045u). However, these same authors describe the difficulty of obtaining this pure pigment (cf. Cai, Sun & Corke, Table 1).
[0025] On the other hand, in silico studies describe tryptophan-betaxanthin as a potential inhibitor of the Sirt1 protein (silent information regulator 1) (Chen et al., Journal of Biomolecular Structure and Dynamics, 2012. DOI: 10.1080 / 07391102.2012.726191). Silent information regulator 1 (Sirt1) is a class III nicotinamide dinucleotide adenine-dependent histone deacetylase. The information provided is restricted to computational molecular modeling studies, without any experimental measure or access to the pure molecule and without giving biological evidence of any potentially positive activity.
[0026] Finally, a study carried out by molecular dynamic simulation "in silico" suggests that tryptophan-betaxanthin may be an agonist of PPAR (Chen et al., Journal of Biomolecular Structure and Dynamics, 2012. DOI: 10.1080 / 07391102.2012.726191), since obtained the highest coupling score, in silico, with respect to the other molecules tested computationally, but, without experimental evidence, of any effect mediated by PPAR.
[0027] BRIEF DESCRIPTION OF THE INVENTION
[0029] One aspect of the present invention relates to a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:
[0033] in which,
[0035] - R ¹ is H or -COOH, or -COOR ⁶ , in which R ⁶ is a C ¹ -C ⁶ alkyl group;
[0036] - R ² is an unsubstituted C ¹ -C ¹⁰ alkyl group or a C ¹ -C ¹⁰ alkyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -R ⁴ -COR ⁵ and -R ⁴ -COOR ⁵ , wherein R ⁴ is a C ¹ -C ⁶ alkyl group and R ⁵ is -H or a C ¹ -C ⁶ alkyl group;
[0037] - R ³ is selected from:
[0038] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR ⁶ , -COOH and -COOR ⁶ , wherein R ⁶ is a C ¹ -C ⁶ alkyl group;
[0039] or
[0040] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR ⁶ , -COOH and -COOR ⁶ , where R ⁶ is a C ¹ -C ⁶ alkyl group;
[0042] and when R ³ is an unsubstituted phenyl group, R ¹ is H;
[0044] for use in the treatment and / or prevention of cancer and tumors in humans and animals, as sole treatment or accompanying other therapeutic and / or preventive treatments.
[0045] Another aspect of the invention refers to a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:
[0047] in which,
[0048] - Ri is H or -COOH;
[0049] - R ² is an unsubstituted C ¹ -C ¹⁰ alkyl group;
[0050] - R ³ is selected from:
[0051] an unsubstituted phenyl group;
[0052] or
[0053] an unsubstituted 1H-indol-3-yl group;
[0055] and when R ³ is an unsubstituted phenyl group, R ¹ is H;
[0057] for use in the treatment and / or prevention of cancer and tumors in humans and animals, as sole treatment or accompanying other therapeutic and / or preventive treatments.
[0058] Still another aspect of the present invention relates to a pharmaceutical composition comprising the compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof, described in the present invention and, at least one pharmaceutically acceptable excipient, for use in the treatment and / or prevention of cancer and tumors in humans and animals, as sole treatment or accompanying other therapeutic and / or preventive treatments.
[0060] Finally, another aspect of the present invention refers to the use of a nutraceutical or food composition comprising a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof, described in the present invention and, at least a food-acceptable excipient, for cancer prevention or as a chemopreventive agent.
[0061] BRIEF DESCRIPTION OF THE FIGURES
[0063] Figure 1: Chemical structure of betalains with a net positive charge.
[0065] Figure 2: Chemical structure of betalains with no net positive charge, made up of an amine linked to an aromatic ring such as tryptophan-betaxanthine, phenylethylamine betaxanthine, phenylalanine-betaxanthine or dopaxanthin and those that come from a linear amine such as vulgaxanthin I or humilixanthin.
[0067] Figure 3: In vitro antioxidant activity of the compounds described in the present description in comparison with bethalamic acid.
[0069] Figure 4: In vivo antioxidant activity of the compounds described in the present description. The assessment was carried out by measuring the ability to reduce the fluorescence of each compound in the pharynx of the line TJ375 of the nematode C. elegans, as a measure of oxidative stress, when it is exposed to juglone. ( A ) Histogram of antioxidant activity in vivo, ( B ) Representative image of the control exposed to juglone, ( C ) Representative image of an animal treated with 25 µM of dopaxanthin and exposed to juglone.
[0071] Figure 5: In vivo tumor size measurement method: ( A ) C. elegans wild-type phenotype (wild-type, N2), with normal gonad. ( B ) C. elegans mutant phenotype JK1466 with tumor in the gonad. Both specimens have the gonad marked from the loop region to the proximal region. Scale bar 100 ^ m. This measure is used in the evaluation of tumor shrinkage by bioactive compounds.
[0073] Figure 6: Histogram of the areas of the gonads in animals with the wild type phenotype (N2), untreated tumor phenotype (JK1466) and treated with tryptophan-betaxanthine, phenylethylamine betaxanthine, phenylalanine-betaxanthine and dopaxanthin. ** Statistically significant changes.
[0075] Figure 7 : Histograms and survival curves of C. elegans JK1466 control (untreated) and treated with tryptophan-betaxanthine ( A and B ) and phenylethylamine-betaxanthine ( C and D ). ** Statistically significant changes.
[0077] DESCRIPTION
[0079] The present invention relates to a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:
[0081] in which,
[0082] - Ri is H or -COOH, or -COOR6, in which R6 is a C ¹ -C ⁶ alkyl group;
[0083] - R ² is an unsubstituted C ¹ -C ¹⁰ alkyl group or a C ¹ -C ¹⁰ alkyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -R ⁴ -COR ⁵ and -R ⁴ -COOR ⁵ , wherein R ⁴ is a C ¹ -C ⁶ alkyl group and R ⁵ is -H or a C ¹ -C6 alkyl group;
[0084] - R ³ is selected from:
[0085] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0086] or
[0087] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0089] and when R ³ is an unsubstituted phenyl group, R ¹ is H;
[0091] for use in the treatment and / or prevention of cancer and tumors in humans and animals, as the sole treatment or accompanying other therapeutic and / or preventive treatments.
[0092] R ^{2 is} preferably an unsubstituted C ¹ -C ¹⁰ alkyl group. Most preferably R ² is -CH ² -. Also preferably R ³ is selected from:
[0094] an unsubstituted phenyl group or a phenyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
[0095] or
[0096] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group.
[0098] Also, preferably R ³ is selected from:
[0099] an unsubstituted phenyl group;
[0100] or
[0101] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group.
[0103] Also, preferably R ³ is selected from:
[0105] an unsubstituted phenyl group;
[0106] or
[0107] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group.
[0108] Also, preferably R ³ is selected from:
[0110] an unsubstituted phenyl group or a phenyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
[0111] or
[0112] an unsubstituted 1H-indol-3-yl group.
[0114] More preferably the C ¹ -C ⁶ alkyl group is methyl or ethyl.
[0116] Also more preferably R ³ is selected from:
[0117] an unsubstituted phenyl group;
[0118] or
[0119] an unsubstituted 1H-indol-3-yl group.
[0121] Thus, the present invention also relates to said use in the case of the enantiomers of the compound of formula (I), which have the opposite stereochemical configuration in relation to the two chiral centers marked with an asterisk (*).
[0123] Likewise, the present invention also relates to said use in the case of the diasteromers of the compound of formula (I), which have the opposite stereochemical configuration in relation to only one of the two chiral centers marked with an asterisk (*).
[0124] Finally, the present invention also relates to said use of all the E / Z isomers of the compound of formula (I) in relation to the configuration of the bond substituents double carbon-carbon.
[0126] Due to the activity of the compounds of formula (I), they can be used for the treatment and / or prevention of cancer in pharmaceutical compositions, but also as chemopreventive agents and in cancer prevention, in nutraceutical and / or food compositions.
[0128] The compounds of formula (I) exhibit antioxidant activity, as well as antitumor activity.
[0130] Specifically, the present invention shows the antitumor effects of the compounds of formula (I), exemplified by tryptophan-betaxanthine and phenylethylamine-betaxanthine in vivo in the Caenorhabditis elegans (nematode) animal model (Figure 6). It should be noted that until now it has not been possible to carry out in-depth studies such as those presented by the invention since the levels of tryptophan-betaxanthin in plants are very low and not feasible for an in vivo study . For its synthesis, the biotechnological method described by Guerrero-Rubio et al., (Guerrero-Rubio et al., Microbial Biotechnology, 2019. DOI: 10.1111 / 1751-7915.13452) has been used. Thus, each of the betaxanthines of the invention have been individually evaluated, carrying out an in-depth study of their antitumor effects in an animal model.
[0131] More specifically, this invention describes the antitumor and antiproliferative effect of the compounds of formula (I) using the animal model C. elegans, specifically the mutant line JK1466 (Figures 5 and 6). This strain of C. elegans shows a mutation of the gld-1 tumor suppressor gene . In gld-1 (q485) mutants, germ cells are unable to emerge from mitosis and continue to proliferate throughout the gonad, forming a germline tumor that is lethal to the animal (Francis et al., Genetics, 1995, 139 (2): 579 606). An objective evaluation method of tumor growth in this line has been developed to evaluate the possible antitumor activity of the molecules of the invention, of imine structure, pure in vivo.
[0133] Furthermore, to confirm the existence or not of a direct relationship between the antioxidant capacity of the compounds of formula (I) of the invention and their antitumor effects, their antioxidant capacity has been assessed both in vitro and in vivo (Figures 3 and 4 ).
[0135] The in vitro assessment was done by the ABTS method (Re et al., Free radical biology and medicine, 1999. DOI: 10.1016 / S0891-5849 (98) 00315-3). The in vivo assessment was carried out with the mutant TJ375 strain of C. elegans , this strain has fused the GFP protein to the heat shock proteins, which is expressed in the pharynx of C. elegans (Guerrero-Rubio et al., FoodChemistry, 2019. DOI: 10.1016 / j.foodchem.2018.09.067). The animals are treated with the compound to be evaluated and after that they are exposed to oxidative stress produced with juglone. (naphthoquinone). The lower the activation of heat shock proteins, the higher the antioxidant capacity of the compound. The oxidative stress state of the animal is evaluated by the area and intensity of fluorescence associated with the accumulation of fluorescent HSP.
[0137] Betalains with uncharged imine structure have been used that have an aromatic ring in their structure, obtained from the corresponding primary amine, such as: tryptophanobetaxanthine, phenylethylamine-betaxanthine, phenylalanine-betaxanthine and dopaxanthin, which were synthesized through a biotechnological process. Biotechnological production was carried out in microbial factories following a method previously described by Guerrero-Rubio et al., Microbial Biotechnology, 2019 (DOI: 10.1111 / 1751-7915.13452).
[0139] Although these molecules have a similar structure, small structural differences were decisive for obtaining antioxidant and antitumor activity.
[0140] Thus, this invention shows experimentally that the compounds of formula (I), exemplified with tryptophan-betaxanthine and phenylethylamine-betaxanthine, have a health-promoting effect and more specifically an antitumor effect, without having any direct relationship with their antioxidant capacity.
[0142] Specifically, dopaxanthin was the molecule with the greatest capacity to reduce oxidative stress in vivo, but it did not show any antitumor capacity. However, tryptophan-betaxanthin was the molecule with the least capacity to reduce oxidative stress in vivo, but it was the molecule that showed the greatest efficacy in reducing tumor size in C. elegans and a possible candidate in new treatment strategies and / or prevention of diseases such as cancer.
[0144] The state of the art (ES2349522B2, or Khan et al., FTW-Food Science and Technology, 2012. DOI: 10.1016 / j.lwt.2012.01.025) suggests that the antioxidant capacity of betalains is linked to the presence of H donor groups, such as the imino groups that all of them have, or a greater number of hydroxyl groups ( cf. ES2349522B2 page 3 lines 2-6 and Khan et al., page 320 second column, last lines). However, it seems that the state of the art is contradictory, indicating the patent ES2349522 B2 that betaxanthins exhibit a greater antioxidant capacity than betacyanins ( cf. ES2349522 B2, page 3 lines 2-3), while Farabegoli et al. describe the opposite ( cf. Farabegoli et al., page 359, column 2, last paragraph of section 3.1).
[0146] Furthermore, as indicated above, it cannot be generalized that a molecule because it has antioxidant effects in vitro or even inferred because it is structurally similar to another is effective as an antitumor.
[0147] The presence of hydroxyl, as suggested by both Khan et al., And patent ES2349522 B2, does not seem to help to select those betalains with anticancer activity, since the phenylethylamine-betaxanthine compound, for use in the treatment and / or prevention of Cancer, according to the present invention, shows antitumor activity, while dopaxanthin, which has a higher number of hydroxyls, does not show such activity.
[0148] A preferred embodiment of the invention relates to the use of a compound of formula (I), in which:
[0149] - R ¹ is -COOH;
[0150] - R ² is -CH ² -; Y
[0151] - R ³ is selected from:
[0152] a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
[0153] or
[0154] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0155] and the compound has formula (II):
[0159] Preferably R ³ is selected from:
[0160] a phenyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0161] or
[0162] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR ⁶ , -COOH and -COOR ⁶ , where R ⁶ is a C ¹ -C ⁶ alkyl group.
[0164] Also preferably, R ³ is selected from:
[0166] a phenyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR ⁶ , -COOH and -COOR ⁶ , wherein R ⁶ is a C ¹ -C ⁶ alkyl group;
[0167] or
[0168] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR ⁶ , -COOH and -COOR ⁶ , where R ⁶ is a C ¹ -C ⁶ alkyl group.
[0170] Also preferably, R ³ is selected from:
[0171] a phenyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR ⁶ , -COOH and -COOR ⁶ , wherein R ⁶ is a C ¹ -C ⁶ alkyl group;
[0172] or
[0173] an unsubstituted 1H-indol-3-yl group.
[0175] A more preferred embodiment refers to a compound of formula (II) for use according to the present invention, in which R ³ is an unsubstituted 1H-indol-3-yl group, and the compound is tryptophan-betaxanthin:
[0179] Another preferred embodiment refers to a compound of formula (I) for use according to the present invention, in which:
[0180] - R ¹ is H;
[0181] - R ² is -CH ² -; Y
[0182] - R ³ is selected from
[0183] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0184] or
[0185] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0187] and the compound has formula (III):
[0191] R ³ is preferably selected from:
[0193] an unsubstituted phenyl group;
[0194] or
[0195] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group.
[0197] Also preferably, R ³ is selected from:
[0198] an unsubstituted phenyl group;
[0199] or
[0200] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group.
[0202] Another more preferred embodiment refers to a compound of formula (III) for use according to the present invention, wherein R ³ is an unsubstituted phenyl group, and the compound is phenylethylamine-betaxanthine:
[0205] Thus the present application refers to the use of a compound of formula (I) or of an enantiomer, diasteromer or E / Z isomer thereof, more preferably to the use of a compound of formula (II) or of a compound of formula (III ), even more preferably to the use of tryptophanobetaxanthine or phenylethylamine-betaxanthine, for the treatment and / or prevention of cancer and tumors in humans and animals, as a single treatment or accompanying other therapeutic and / or preventive treatments.
[0207] Furthermore, another aspect of the invention relates to a pharmaceutical composition comprising a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:
[0211] in which,
[0212] - R ¹ is H or -COOH, or -COOR ⁶ , in which R ⁶ is a C ¹ -C ⁶ alkyl group;
[0213] - R ² is an unsubstituted C ¹ -C ¹⁰ alkyl group or a C ¹ -C ¹⁰ alkyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -R ⁴ -COR ⁵ and -R ⁴ -COOR ⁵ , wherein R ⁴ is a C ¹ -C ⁶ alkyl group and R ⁵ is -H or a group C ¹ C ⁶ alkyl;
[0214] - R ³ is selected from:
[0215] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0216] or
[0217] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0219] and when R ³ is an unsubstituted phenyl group, R ¹ is H;
[0221] and at least one pharmaceutically acceptable excipient, for use in the treatment and / or prevention of cancer and tumors in humans and animals.
[0223] An effective amount, for the purposes of the present invention, is understood as that which provides a therapeutic effect without providing unacceptable toxic effects in the patient. The effective amount or dose of the drug depends on the compound and the condition or disease treated and on, for example, the age, weight and clinical condition of the patient treated, the method of administration, the clinical history of the patient, the severity of the disease. and the potency of the administered compound.
[0225] Compositions for use according to the present invention can comprise one or more compounds of formula (I), and also one or more active compounds. For the purposes of the present invention, an active compound is understood to be a chemical entity or a molecule that exerts therapeutic effects when administered to a human or animal.
[0226] A preferred embodiment refers to a pharmaceutical composition for use according to the present invention, which comprises a compound of formula (I) in which:
[0228] - R ¹ is -COOH;
[0229] - R ² is -CH ² -; Y
[0230] - R ³ is selected from:
[0231] a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
[0232] or
[0233] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0234] and the compound has formula (II):
[0238] A more preferred embodiment relates to a pharmaceutical composition for use according to the present invention, comprising a compound of formula (II) in which R ³ is an unsubstituted 1 H-indol-3-yl group, and the compound is tryptophan-betaxanthin:
[0242] Another preferred embodiment refers to a pharmaceutical composition for use according to the present invention, comprising a compound of formula (I) in which:
[0243] - R ¹ is H;
[0244] - R ² is -CH ² -; Y
[0245] - R ³ is selected from
[0246] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0247] or
[0248] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0249] and the compound has formula (111):
[0253] Another more preferred embodiment refers to a pharmaceutical composition for use according to the present invention, which comprises a compound of formula (III) in which R ³ is an unsubstituted phenyl group, and the compound is phenylethylamine-betaxanthine:
[0257] Compositions for use according to the present invention include, together with the compounds described in the present invention at least one pharmaceutically acceptable excipient, which may be, inter alia, a carrier or diluent.
[0259] Said compositions can be included in capsules, tablets, sachets or sachets or any other type of presentation.
[0261] To make such compositions, conventional techniques for the preparation of pharmaceutical compositions can be used. For example, the compound of interest can be mixed with a vehicle or diluted in a vehicle or contained in a vehicle in the form of an ampoule, capsule, tablet, sachet, sachet or other container. When the vehicle serves as a solvent, it can be solid, semi-solid or liquid and act as an excipient or medium for said active compound. The compound of interest can be adsorbed onto a solid granular medium. Some examples of suitable vehicles are water, saline solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, lactose, terra alba, sucrose, cyclodextrins, amylose, magnesium stearate, talc, gelatin, agar, pectin , acacia, stearic acid, cellulose alkyl ethers, silicon acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythrole fatty esters, polyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. Likewise, the vehicle or support can include sustained release materials known in the state of the art, such as glyceryl monostearate or diesterate alone or mixed with a wax. The formulations can also include wetting, emulsifying, suspending, preserving, sweetening or flavoring agents. The compositions can be formulated to provide rapid, sustained or delayed release of the active agent after it is administered to the patient using methods known in the art.
[0263] The pharmaceutical compositions can be sterilized and mixed, if desired, with additional agents, emulsifiers, salt to influence osmotic pressure, buffers and / or coloring substances that do not react adversely with the active compounds.
[0264] One embodiment refers to the mode of administration, which can be any mode that effectively transports the compound of interest to the desired site of action, such as oral, rectal, or parenteral, for example, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, or as an ophthalmic solution.
[0266] Preferably, the pharmaceutical composition for use according to the present invention is a tablet, an injectable solution, or an ophthalmic solution.
[0268] For oral administration, both solid and liquid dosage forms can be prepared. To prepare solid compositions as tablets, the compound of interest is mixed in a formulation with other conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, starch, lactose, acacia, methyl cellulose and functionally similar materials such as pharmaceutical carriers and diluents.
[0270] Capsules can be prepared by mixing the compound of interest with a pharmaceutically inert solvent and filling the mixture into an appropriately sized hard gelatin. Soft capsules are prepared with machines for encapsulating suspensions of the compound of interest with an acceptable vegetable oil, a light paraffin or an inert oil.
[0271] Liquid dosage forms such as syrups, elixirs and suspensions can also be prepared. The water-soluble forms can be dissolved in an aqueous vehicle along with sugar, flavorings, and preservatives to form a syrup. An elixir is prepared using a hydroalcoholic vehicle (eg ethanol) with suitable sweeteners such as sugar or saccharin, along with flavoring aromatic agents. Suspensions can be prepared with an aqueous vehicle and the aid of a suspending agent such as acacia, tragacanth, methylcellulose, and the like.
[0273] For nasal administration, the preparation may contain the compound of interest dissolved or suspended in a liquid vehicle, in particular an aqueous vehicle, for application as an aerosol. The carrier may contain additives such as solubilizing agents, for example propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives such as parabens.
[0275] For ophthalmic applications, the compound of interest is formulated in solutions, suspensions and ointments suitable for use in the eye. The concentrations are usually the same as in preparations for local use.
[0277] For parenteral application, the use of injectable solutions or suspensions, for intradermal, intramuscular, intravascular and subcutaneous use, are obvious to the person skilled in the art.
[0278] In addition to the compound of interest, the compositions may include other pharmaceutically acceptable non-toxic diluents and excipients, including vehicles commonly used in pharmaceutical compositions commonly used in humans or animals. The diluent is selected so that it does not affect the biological activity of the composition.
[0280] Examples of diluents used especially in injectable formulations are organic and inorganic saline solutions, Ringer's solution, dextrose solution and Hank's solution. In addition, the compositions can include additives such as other excipients, adjuvants, non-therapeutic and non-immunogenic stabilizers, and the like.
[0282] Examples of excipients that can be included in the formulation include, but are not limited to cosolvents, surfactants, oils, humectants, emollients, preservatives, stabilizers, and antioxidants. Any physiologically acceptable buffer can be used, such as Tris or phosphate buffers. Effective amounts of diluents or additives or excipients are those that are effective to obtain a pharmaceutically acceptable formulation in terms of solubility and biological activity.
[0284] Another embodiment relates to the dosage regimen. The term "unit dose" refers to physically discrete units suitable as unit doses for an individual, eg, mammal, human, dog, cat, rodent, etc. where each unit contains one predetermined amount of active material calculated to produce the appropriate therapeutic effect in association with the appropriate diluent, carrier or vehicle.
[0286] A preferred embodiment refers to a pharmaceutical composition for use according to the present invention, wherein said composition is administered by oral, intramuscular, intravenous, subcutaneous, inhalation, transdermal, nasal, ophthalmic, otic, topical, rectal or vaginal administration. .
[0288] Another embodiment refers to a method of treatment and / or prevention of cancer and tumors in humans and animals, which comprises the administration of an effective amount of a compound of formula (I), or enantiomer, diasteromer or E / Z isomer thereof. , or of a pharmaceutical composition comprising it.
[0290] Since the compounds of formula (I) can also exert cancer prevention effects and be used as chemopreventive agents, they can also form nutraceutical or food compositions together with other compounds, which can also be used in cancer prevention.
[0292] In particular, the compounds of formula (I) can also be used as chemopreventive agents, or to improve the health of the cancer patient.
[0294] The term "chemopreventive agent" refers to a compound or substance whose administration delays or prevents the onset of cancer.
[0296] Thus, the present invention also relates to a compound of formula (I) or to an enantiomer, diasteromer or E / Z isomer thereof, more preferably to the use of a compound of formula (II) or of a compound of formula (III) , even more preferably to the use of tryptophan-betaxanthine or phenylethylamine-betaxanthine, for use as a chemopreventive agent, or to improve the health of the cancer patient
[0298] Thus, another aspect of the invention refers to the use of a nutraceutical or food composition comprising a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:
[0300] in which,
[0301] - Ri is H or -COOH, or -COOR6, in which R6 is a C ¹ -C ⁶ alkyl group;
[0302] - R ² is an unsubstituted C ¹ -C ¹⁰ alkyl group or a C ¹ -C ¹⁰ alkyl group substituted with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -R ⁴ -COR ⁵ and -R ⁴ -COOR ⁵ , wherein R ⁴ is a C ¹ -C ⁶ alkyl group and R ⁵ is -H or a C ¹ -C6 alkyl group;
[0303] - R ³ is selected from:
[0304] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0305] or
[0306] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0308] and when R ³ is an unsubstituted phenyl group, R ¹ is H;
[0310] and at least one food-acceptable excipient, for the prevention of cancer or as a chemopreventive agent.
[0312] The term "food acceptable excipient" refers, for the purposes of the present invention, to a carrier or diluent.
[0314] For the purposes of the present invention, a nutraceutical composition is understood to be a food composition, to be ingested separately or with food, which has a medicinal effect on human health.
[0316] In a preferred embodiment, the nutraceutical composition for use according to the invention comprises a compound of formula (I) in which:
[0317] - Ri is -COOH;
[0318] - R ² is -CH ² -; Y
[0319] - R ³ is selected from:
[0320] a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
[0321] or
[0322] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0323] and the compound has formula (II):
[0327] In a more preferred embodiment, the nutraceutical composition for use according to the invention comprises a compound of formula (II) in which R ³ is an unsubstituted 1H-indol-3-yl group, and the compound is tryptophan-betaxanthin:
[0331] In another preferred embodiment, the nutraceutical composition for use according to the invention comprises a compound of formula (I) in which:
[0332] - R ¹ is H;
[0333] - R ² is -CH ² -; Y
[0334] - R ³ is selected from
[0335] an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
[0336] or
[0337] an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
[0338] and the compound has formula (III):
[0342] In another more preferred embodiment the nutraceutical composition for use according to the invention comprises a compound of formula (III) in which R ³ is an unsubstituted phenyl group, and the compound is phenylethylamine-betaxanthine:
[0343] In another preferred embodiment of the nutraceutical composition for use according to the invention, the nutraceutical composition is incorporated into a food preparation.
[0345] The nutraceutical composition for use according to the invention can also be included in a variety of food preparations, for example, products derived from milk, yogurt, curd, cheese (for example, fresh, cream, processed, soft and hard cheese), fermented milk , powdered milk, a fermented milk-based product, an ice cream, a fermented cereal-based product, a milk-based powder, drinks, and a pet food.
[0346] The term "food preparations" is used here in its broadest sense, including any type of product, in any form of presentation, that can be ingested by an animal, but with the exception of pharmaceutical and veterinary products. Examples of other food preparations are meat products (e.g. pâtés, frankfurters and salami sausages or meat spreads), chocolate spreads, fillings (e.g. truffle, cream) and glazes, chocolate, confectionery (e.g. caramel, fondant or toffee), bakery products (cakes, pasta), sauces and soups, fruit juices and coffee whiteners. Particularly interesting food preparations include food supplements and infant formulas.
[0348] The nutraceutical composition for use according to the invention could also be used as an ingredient in other food products. Accordingly, in another aspect of the invention, food preparations are provided containing the composition of the invention together with appropriate amounts of edible ingredients. Preferably, the nutraceutical composition for use according to the invention is a food supplement. For the purposes of the present invention, the term "food supplement" refers to that fraction of food that is used to complete human or animal nutrition. If the nutraceutical composition for use according to the invention is used as a food supplement, it can be administered as such, or it can be mixed with a suitable drinking liquid, such as water, yogurt, milk or fruit juice, or it can be mixed with solid or liquid foods. . In this context, the food supplement may be in the form of tablets, pills, capsules, granules, powders, suspensions, sachets, lozenges, candies, bars, syrups, and corresponding forms of administration, generally in unit dose form.
[0350] EXAMPLES
[0352] The examples described below are illustrative in nature and are not intended to limit the scope of the present invention.
[0353] Example 1: Biotechnological obtaining of compounds of formula (I).
[0355] To obtain the compounds of formula (I), the method published by Guerrero-Rubio et al ., 2019 ( Microbial Biotechnology. DOI: 10.1111 / 1751-7915.13452) was used.
[0357] Bethalamic acid was produced by a novel and efficient DODA (4,5-DOPA-extradiol-dioxygenase) enzyme from Gluconacetobacter and then condensed with excess amines or individual amino acids in cell factories to obtain betaxanthins. Four structure betalains were produced with no net positive charge. For this, cell cultures containing the enzyme DODA, distilled water, 7.6 mM L-Dopa (L-3,4-dihydroxyphenylalanine), 15 mM sodium ascorbate and additionally the amino acids or amines 38 mM L- were added to a flask. tryptophan, L-phenylethylamine and L-phenylalanine to form the compounds of formula (I), L-tryptophan-betaxanthine and L-phenylethylamine-betaxanthine, as well as the compounds L-phenylalanine-betaxanthine, and dopaxanthin. All this while stirring and at 20 ° C for 48 h.
[0359] These betalains of imine structure were purified to homogeneity by anion exchange chromatography and solid phase extraction following a previously described method (García-Herrero, Escribano & García-Carmona, Planta, 2010. DOI: 10.1007 / s00425-010-1191-0 ; Guerrero-Rubio et al, Microbial Biotechnology, 2019. DOI: 10.1111 / 1751-7915.134522019).
[0361] Example 2: In vitro evaluation of the antioxidant capacity.
[0363] The antioxidant capacity of each of the molecules was evaluated using the ABTS method, this technique reveals whether an antioxidant compound is capable of reducing the ABTS radical to ABTS expressed in TEAC (Trolox equivalent antioxidant capacity). As can be seen in Figure 3, dopaxanthin is the most antioxidant in vitro (6.8 TEAC), as expected due to the presence of the catechol substructure (Gandía-Herrero et al., Journal of Natural Products, 2009. DOI: 10.1021 / np900131r; Gandía-Herrero et al., Planta, 2010. DOI: 10.1007 / s00425-010-1191-0). Values for tryptophan-betaxanthine (5.8 TEAC) and phenylalanine-betaxanthine (2.6 TEAC) and phenylethylamine betaxanthine (2.5 TEAC), which have an antioxidant capacity very similar to beta-thalamic acid (2.7 TEAC), were also determined. . Therefore, the results show that the molecule with the highest antioxidant capacity is dopaxanthin.
[0365] Example 3. In vivo evaluation of the antioxidant capacity.
[0367] The ability to eliminate free radicals in vivo was carried out by the lineage of the nematode C. elegans TJ375, this lineage has fused to the heat shock protein HSP-16.2 the green fluorescent protein GFP, which when the animal is subjected to stress oxidative is added in the pharynx of animals (Figure 4 BC) and its accumulation can be assessed by fluorescence microscopy. In Figure 4 A and C it can be seen how dopaxanthin is also the major antioxidant in vivo since it reduces oxidative stress in the animal by 84%. Phenylalanine-betaxanthine and phenylethylamine-betaxanthine are also capable of greatly reducing oxidative stress in animals since the accumulation of fluorescence is reduced by 68.9% and 64.9% respectively. The tryptophanobetaxanthin molecule is the least effective in reducing oxidative stress (23.2%). That is, the tryptophan-betaxanthin molecule is not a good antioxidant in vivo.
[0369] Example 4. In vivo evaluation of antitumor capacity.
[0371] Each pure betaxanthin was tested individually. The mutant strain of C. elegans JK1466 (tumor phenotype) was treated with 25 µM of each of the molecules obtained in a standardized culture medium of C. elegans (Medium S). The treatment was carried out during four adult days and on the fourth day the size of the tumors was analyzed. Taking into account the results of the bibliography and if the works that suggest a direct relationship between antioxidant capacity and antitumor or anticancer activity are considered good, it is to be expected that, being dopaxanthin the molecule with the highest antioxidant capacity of all those tested, this will be the more effective in treating tumors.
[0373] To examine the effect of pure betaxanthins on tumor growth, the sizes of the gonads were measured from the loop region to the proximal region (Figure 5), including the area of the uterus when it was filled with tumor cells. None of the pigments showed a growth response in tumor size.
[0375] Unexpectedly, the greatest effect on tumor reduction was caused by tryptophanobetaxanthine, which reduced the tumor by 56.4% taking as a starting point the size of the gonad of the wild type phenotype (N2) (Figure 6), followed by phenylethylamine- betaxanthin that reduced the tumor by 27.7% (Figure 6).
[0377] However, phenylalanine-betaxanthin and dopaxanthin did not have a significant effect on C. elegans tumor size (Figure 6) although they are potent antioxidants in vivo as shown in Figure 3. These data demonstrate that nematodes that are fed individually with phenylethylamine-betaxanthine and especially with tryptophanobetaxanthine, significantly reduced tumor growth. Thus, the fact that they are antioxidant and anti-radical molecules, as are all betalains (including dopaxanthin and phenylalanine-betaxanthin), does not mean that they are anti-tumor. Therefore, it was not predictable that the most active molecules in tumor reduction were tryptophanobetaxanthine and phenylethylamine-betaxanthine.
[0378] Example 5. Effect on the longevity of the tumor C. elegans model.
[0380] Among the four betalains of imine structure, two of them were selected due to their effect on the inhibition of tumor growth, and survival tests were carried out with phenylethylamine-betaxanthine and tryptophan-betaxanthine, in the line of C. elegans JK1466. The half-life analysis was performed on the automatic platform based on the Lifespan machine (Stroustrup et al., Nature Methods, 2013. DOI: 10.1038 / nmeth.2475). The results show a significant increase in the half-life of C. elegans treated with phenylethylamine-betaxanthine, where the average survival time increased from 8.2 to 9.1 days (Figure 7 C and D). Therefore, the mean survival time increased by 11.4%. Tryptophan-betaxanthin also significantly increased median survival by 9.3%, from 8.2 to 8.9 days (Figure 7 A and B). All mean survival increases determined are statistically significant ( p <0.05). Therefore, phenylethylamine-betaxanthine and tryptophanobetaxanthine not only reduced tumor growth, but also increased the lifespan of C. elegans significantly.
[0382] The results collected for the present invention show the efficacy of tryptophanobetaxanthin as an antitumor treatment, a result that was not expected since the data obtained on antioxidant activity both in vitro and in vivo postulated dopaxanthin as the most effective antitumor treatment because it is the most effective. antioxidant. These results indicate that, although the antioxidant capacity that would have a generic effect on free radicals may in some cases be favorable, as stated in the bibliography, for the effective treatment of tumors it is necessary for the compound to have effects related to the singular structure of each molecule on the tumor as occurs with tryptophan-betaxanthine and phenylethylamine-betaxanthine.

权利要求:
Claims (17)
[1]
1. A compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:

[2]
2. A compound for use according to claim 1, wherein:
- R ¹ is -COOH;
- R ² is -CH ² -; Y
- R ³ is selected from:
a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
or
an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
and the compound has formula (II):

[3]
3. A compound for use according to claim 2, wherein R ³ is an unsubstituted 1H-indol-3-yl group, and the compound is tryptophan-betaxanthine:

[4]
4. A compound for use according to claim 1, wherein:
- R ¹ is H;
- R ² is -CH ² -; Y
- R ³ is selected from
an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, Ci-C6 alkyl, -OR6, -COOH and -COOR6, in which R6 is a C group ¹ -C ⁶ alkyl;
or
an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
and the compound has formula (111):

[5]
5. A compound for use according to claim 4, wherein R ³ is an unsubstituted phenyl group, and the compound is phenylethylamine-betaxanthine:

[6]
6. A pharmaceutical composition comprising an effective amount of the compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:

[7]
7. A pharmaceutical composition for use, according to claim 6, for oral, intramuscular, intravenous, subcutaneous, inhalational, transdermal, nasal, ophthalmic, otic, topical, rectal or vaginal administration.
[8]
8. A pharmaceutical composition for use, according to any of claims 6 or 7, wherein:
- Ri is -COOH;
- R ² is -CH ² -; Y
- R ³ is selected from:
a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ^{6 group} I rent;
or
an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
and the compound has formula (II):

[9]
9. A pharmaceutical composition for use, according to claim ⁸ , in which R ³ is an unsubstituted 1 H-indol-3-yl group, and the compound is tryptophan-betaxanthin:
[10]
10. A pharmaceutical composition for use, according to any of claims 6 or 7, wherein:
- R ¹ is H;
- R ² is -CH ² -; Y
- R ³ is selected from
an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C6 alkyl, -ORa, -COOH and -COOR6, wherein R6 is a group C ¹ -C ⁶ alkyl;
or
an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and - COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
and the compound has formula (111):

[11]
11. A pharmaceutical composition for use, according to claim 10, wherein R ³ is an unsubstituted phenyl group, and the compound is phenylethylamine-betaxanthine:
[12]
12. Use of a nutraceutical composition comprising a compound of formula (I), or an enantiomer, diasteromer or E / Z isomer thereof:

[13]
13. Use according to claim 12, wherein:
- R ¹ is -COOH;
- R ² is -CH ² -; Y
- R ³ is selected from:
a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
or
an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
and the compound has formula (II):

[14]
14. Use according to claim 13, in which R ³ is an unsubstituted 1 H-indol-3-yl group, and the compound is tryptophan-betaxanthin:

[15]
15. Use according to claim 12, wherein:
- R ¹ is H;
- R ² is -CH ² -; Y
- R ³ is selected from:
an unsubstituted phenyl group or a phenyl group substituted with a single OH group or with one or more substituents independently selected from: halogen, Ci-C6 alkyl, -OR6, -COOH and -COOR6, wherein R6 is a C ¹ -C ⁶ alkyl group;
or
an unsubstituted 1H-indol-3-yl group or a 1H-indol-3-yl group substituted with one or more substituents independently selected from: OH, halogen, C ¹ -C ⁶ alkyl, -OR6, -COOH and -COOR6, where R6 is a C ¹ -C ⁶ alkyl group;
and the compound has formula (111):

[16]
16. Use according to claim 15, wherein R ³ is an unsubstituted phenyl group, and the compound is phenylethylamine-betaxanthine:

[17]
Use according to any one of claims 12 to 16, in which the nutraceutical composition is incorporated into a food preparation or food supplement.

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同族专利:

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公开号 | 公开日

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WO2021009029A1|2021-01-21|

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ES2802814B2|2021-10-27|

引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

US6312697B1|1996-07-24|2001-11-06|Govind J. Kapadia|Inhibitory effect of synthetic and natural colorants on carcinogenesis|

---

US20030036565A1|2001-06-22|2003-02-20|Wisconsin Alumni Research Foundation|Cancer chemopreventive agents|

---

DE202005002324U1|2005-02-14|2006-06-22|Treusch, Gernot|Pharmaceutical composition for the fight against cancer|

---

ES2349522B1|2009-05-05|2011-10-27|Universidad De Murcia|PROCEDURE FOR OBTAINING BETALAMIC ACID AND USE OF IT.|

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优先权:

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申请号 | 申请日 | 专利标题

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ES201930649A|ES2802814B2|2019-07-12|2019-07-12|BETAXANTHINES DERIVED FROM TRYPTOPHAN AND PHENYLETHYLAMINE FOR USE IN THE TREATMENT AND / OR PREVENTION OF CANCER|ES201930649A| ES2802814B2|2019-07-12|2019-07-12|BETAXANTHINES DERIVED FROM TRYPTOPHAN AND PHENYLETHYLAMINE FOR USE IN THE TREATMENT AND / OR PREVENTION OF CANCER|

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PCT/EP2020/069516| WO2021009029A1|2019-07-12|2020-07-10|Tryptophan- and phenylethylamine-derived betaxanthins for use in the treatment and/or prevention of cancer|