Aldoximes as no donors, and their uses as plant architecture modifiers and in therapy

Abstract

It relates to aldoximes or pharmaceutically acceptable salts thereof or isomers thereof able of releasing nitric oxide (NO) for use in the treatment of a condition or a disease which is sensible to the action of the NO, wherein the treatment comprises administering the aldoxime to a subject and releasing NO from the aldoxime through an in vivo enzymatically catalyzed redox reaction at physiological pH, as well as to the in vivo use of such aldoximes or acceptable salts for plants thereof or isomers thereof for biological applications which are sensible to the action of the NO in plants, wherein the biological application is selected from the group consisting of plant rooting agent, plant structure modifier, stimulators of the tolerance to abiotic or biotic stress, plant protector from stress induced by ammonium nutrition as an exclusive source of nitrogen, plants protector from hydric stress, and as germination inducer. It also relates to compositions comprising them.

Description

[0001] Aldoximes as NO donors, and their uses as plant architecture modifiers and in therapy

[0002] The present invention relates to aldoximes as NO donors, and its uses as plant architecture modifiers, and in therapy for the treatment of several conditions and diseases.

[0003] Background Art

[0004] Nitric oxide (NO) is a free radical of great importance as a signaling agent in plants, and also in animals, where it is known to be the inducing agent of endothelial relaxation (Nobel prize-winning discovery in 1998).

[0005] In animals NO is produced in different cells and tissues by the transformation of the amino acid L-arginine into L-citrulline through the action of the enzyme nitric oxide synthase (NOS). Until today, it is known that NO exerts a crucial role in the relaxation and functioning of arteries and veins, so NO donors have been used to prevent and treat acute cardiac conditions. In addition, NO has also been used in cancer therapy due to its ability to mediate DNA base amination, enzyme and protein nitrosylation, cell dysfunction, inflammatory reactions and apoptosis. It also participates in the immune defense response, against bacterial infections as a signaling agent in the dispersion of bacterial biofilms at low concentrations, or as an inhibitor of DNA repair at high concentrations, as well as in the processes of healing in all its phases, including vasodilation and anti-platelet effects during the inflammatory process, re-epithelialization and angiogenesis during the proliferative phase, and increased collagen deposition during the reconstruction phase (see Ignarro LJ et al.,” Nitric Oxide Donors and Cardiovascular Agents Modulating the Bioactivity of Nitric Oxide”. Circulation Research, 2002. Vol. 9, pp.:21-28; and Cheng et al., “Nitric Oxide (NO)-Releasing Macromolecules: Rational Design and Biomedical Applications”, Frontiers in Chemistry 2019, vol. 7, pp. 530).

[0006] On the other hand, the formation of nitrogen oxides including NO from oxidative cleavage of C=N(OH) bonds is known. For instance, it is known that several compounds such as aldoximes, ketoximes, amidoximes and guanidoximes can be oxidized by liver microsomes from dexamethasone-treated rats with formation of nitrogen oxides such as NO2; NOT and NO. The oxidative cleavage of their C=N(OH) bond appears as a general P450 3A-catalyzed reaction (see Jousserandot Anne et al.," Formation of nitrogen oxides including NO from oxidative cleavage of C=N(OH)"bonds: A general cytochrome P450- dependent reaction", Bioorganic and Medicinal Chemistry Letters 1995, vol. 5, pp. 423- 426). The document explains that nitric oxide had been disclosed as a biological effect or with an important role in the cardiovascular, in the central and peripheral nervous system, and in modulating the immune response. It also discloses that some compounds containing C=N(OH) function act as precursors of nitrogen oxides upon oxidative cleavage of their C=N(OH) bond by rat liver microsomes. Among the compounds tested is benzaldoxime (BenzOx). However, the work by Jousserandot Anne et al (see Table I) shows a discrete formation of nitrogen oxides.

[0007] Furthermore, L. N. Koikov et al., in “Oximes, amidoximes and hydroxamic acids as nitric oxide donors" Mendeleev Communications, 1998, vol. 8, pp. 165-168 analyzed several oximes, amidoximes and hydroxamic acids for their activity as nitric oxide donors. On page 165, Table 1 shows the yields of NO production for different compounds: oximes RCH=NOH compounds 1 and 2; hydroxamic acids RC(=O)=NOH compounds 1 and 2; and hydroxamic acids RC(=O)NOH, compounds 3; and amidoximes RC(NH2)=NOH, compounds 4, in in vitro conditions at a pH =12. For aldoxime 2i with a methyl pyridine substituent, a high percentage of NO release is observed, whereas for aldoxime 1 a-lf, including aldoxime 1b or benzaldoxime (BenzOx compound) the NO release is almost non-existent. Thus, the skilled person from this document learn that oximes are not likely to produce NO in vivo.

[0008] On the other hand, it is known that in plants nitric oxide is a mediator of the response to auxins in the process of adventitious root formation. Nitric oxide modifies and modulates the root structure, increasing the number of secondary roots, it participates in the defense against abiotic or biotic stresses, and enhances nutrient uptake (see Pagnussat Gabriela Carolina et al.; “Nitric oxide is required for root organogenesis” Plant Physiology (Rockville) 2002, vol. 129, pp. 954-956; or Sun Chengliang et al.; “Molecular functions of nitric oxide and its potential applications in horticultural crops” Sun et al., Horticulture Research ( 2021) 8:71).

[0009] The role of oximes in different functions in plant metabolism, either as an end product or as an intermediate in the production of general and specialized metabolites has also been disclosed in the art (see Sorensen Mette et al.; “Oximes: Unrecognized Chameleons in General and Specialized Plant Metabolism” Molecular Plant, 2018, vol. 11 , pp. 95-117)

[0010] However, from what is known in the state of the art, it is derived that there is a need to find exogenous compounds that are NO producers which are of high interest in the treatment of several diseases which are dependent on NO, as well as, for biological applications in plants. Summary of Invention

[0011] The present inventors have found that aldoximes function in vivo as NO producers and NO donors, through a NO release process consisting of a redox reaction catalyzed enzymatically by peroxidases in vivo either at neutral pH or at physiological pH. They have also found that the production of NO from aldoximes is not spontaneous but requires catalysis. Thus, several hemoproteins catalyze the process such as horseradish peroxidase from Armoracia rusticana- (POD) and the presence of flavin enhances NO production. The reaction mechanism establishes that other peroxidases are also useful to catalyze the process, which depends on the oxidase cycle that takes places in its active center. Given that all living organisms present different peroxidases, usually in very high numbers, with plants possessing more than 100 genes per genome (haploid), the aldoximes can be useful in biological applications.

[0012] As it is illustrated in the examples, aldoximes, in particular, 1 H-lndol-3-acetaldoxime (lAOx), 2-(2,3-dihidro-1 H-inden-3-il)-acetaldoxime (IDOx), 1 H-lndol-3-carboxaldoxime (IMOx), 2-,methyl-Butylaldoxime (ButOx), 2-(4-hidroxiphenyl)-acetaldoxime (PhenOx), benzaldoxime (BenzOx), naphtyl-1 -acetaldoxime (NaftOx), 2,4- chlorophenoxyacetaldoxime (2,4-CIOx) and O-methyl-1 H-indole-3-acetaldoxime (IAO- Met), are nitric oxide donors or releasers. They have the following structure:

[0013] Although the compounds show different degrees of effect among them, all of them stimulate the number of primordia that give rise to new lateral roots, as well as the length of the lateral roots, and can also modify other parameters such as thickness (making them thinner) or total root volume. In addition, they also produce a shortening of the main root which together result in a phenotype called super root. The observed effect of the aldoximes (due to their release of NO) is equivalent to the known effect produced by other nitric oxide donors on root structure (Pagnussat et al, 2003), and, therefore, they are useful in biological applications which are sensible to the action of the NO in plants such as a plant rooting agent or a plant structure modifier. In particular, for stimulating lateral root development, and potentiating the root structure of plants.

[0014] This ability to release NO has a great importance in plants but also in animals where the number of peroxidases is also very high. The examples also illustrate that aldoximes affected the growth of cancer cells (MCF-7) in vitro to a greater extent than non-cancerous control cells and, therefore, aldoximes can also be useful in the treatment of a condition or a disease which is sensible to the action of the NO in a subject.

[0015] Accordingly, a first aspect of the present invention relates to an aldoxime able of releasing nitric oxide (NO) for use in the treatment of a condition or a disease which is sensible to the action of the NO, wherein the treatment comprises administering the aldoxime to a subject and releasing NO from the aldoxime through an in vivo, enzymatically catalyzed redox reaction either at neutral or at physiological pH. This means a pH between 7-7.5. The pH can be measured by well-known tests.

[0016] A second aspect of the present invention relates to the use of an aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof able of releasing nitric oxide (NO) to improve the muscle contractility capacity, ventilatory efficiency, and sports performance,

[0017] A third aspect of the present invention relates to an in vivo use of an aldoxime able of releasing nitric oxide (NO) in vivo, for biological applications which are sensible to the action of the NO in plants, wherein the biological application is selected from the group consisting of plant rooting agent, plant structure modifier, stimulators of the tolerance to abiotic or biotic stress, _plants protector from stress induced by ammonium nutrition as an exclusive source of nitrogen, plants protector from hydric stress, and as germination inducer.

[0018] A fourth aspect of the present invention relates to a compound selected from 2-(2,3- dihidro-1 H-inden-3-il) acetaldoxime (IDOx), and 2-(4-hidroxiphenyl)-acetaldoxime, (PhenOx), of formula below:

[0019] PhenOx

[0020] A fifth aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of an aldoxime able of releasing nitric oxide (NO) in vivo, together with pharmaceutically acceptable excipients or carriers.

[0021] A sixth aspect of the present invention relates to a composition for plants comprising an effective amount of an aldoxime able of releasing nitric oxide (NO) in vivo, together with appropriate carriers, wherein the effective amount is for stimulating fruit and root development, in particular, lateral root development i.e. , potentiating the root structure of plants, and for modifying plant structure.

[0022] Brief Description of Drawings

[0023] FIG. 1 shows the production of NO catalyzed by horseradish peroxidase (POD), and detected as fluorescence with the DAF-FM probe, using different aldoximes as substrates (1 H-lndol-3-acetaldoxime, lAOx; 1H-lndol-3-carboxaldoxime, IMOx; 2-(2,3-dihidro-1 H- inden-3-il)-acetaldoxime, IDOx; benzaldoxime, BenzOx; 2-(4-hidroxiphenyl)-acetaldoxime, PhenOx), or different metabolites in the IAA synthetic pathway as IAA (indolil-3-acetic acid), Tryptophane (Trp), N-hidroxyl-tryptamine (NHT)..

[0024] FIG. 2 (a) shows the production of NO as detected using 200 pM indole-3-acetaldoxime (lAOx) as substrate as indicated and 10 pM POD as catalyst, (b) NO production with 200 pM lAOx and different concentrations of the POD catalyst, as indicated. Reactions were performed in 96-well black plates and 0.5 pM 4-Amino-5-methylamino-2',7'-difluorescein (DAF-FM) in 100 mM Tris-HCI pH 7.4 in a total volume of 300pL. The fluorescence measurement was taken 3h after the reaction in a spectrofluorimetry (BioTek Synergy HT) at 485 nm (ex) and 520 nm (em). Values are the mean of 4 replicates ± S.E.

[0025] FIG. 3 shows the iron reduction activity at different pHs for the following aldoximes lAOx, IMOx, BenzOx, and IAA, the last as a control without oxime group. The reduction of Fe3+was measured using the Ferrozin assay. The activity was expressed as the increase of the absorbance at 562 mm after 30’. The values are the media of 3 repetitions ± S.E. Asterisks indicates significant differences (P value <0,001) between molecules and control using ANOVA tests of two factors and Bonferroni.

[0026] FIG. 4 shows the determination by chemiluminescence of NO, nitrite and nitrate produced from lAOx as a result of the catalysis of different types of hemoproteins (either rice Hb1 , Lbl I, or POD. Reactions were performed in sealed glass vials containing 2 mM aldoxime, 10 pM hemoprotein as catalyst, and 100 mM Tris-HCI pH 7.4 in 1 mL. Iron-Superoxide Dismutase (FeSOD) was produced recombinantly according to methodology described (Moran et al., “Functional characterization and expression of a cytosolic iron-superoxide dismutase from cowpea root nodules”, Plant Physiology 2003, vol. 133, pp. 773-782) and it was included in the reactions at final concentration of 1 pM. NO was determined directly and as nitrite and / or nitrate production. Nitrite or nitrate production were analyzed by reducing it to NO in a purge vessel with KI and acetic acid and / or VCI3 and HOI, respectively, since the NO initially produced spontaneously oxidizes to nitrite and nitrate in the presence of oxygen. A NO Analyzer (Serinus 40, Ecotech) was used to analyze by chemiluminiscence the NO. Values are the mean of 5 replicates ± S.E. Group letters indicate significant differences (P value <0.01) in the independent t-test within each hemoprotein; asterisks indicate significant differences (p-value <0.001) in Dunnett's oneway ANOVA and post hoc test using lAOx + POD as control.

[0027] FIG. 5 shows the production of NO from lAOx under anoxia / normoxia or anoxia + superoxide conditions as indicated. The reactions were carried out in airtight glass vials containing 2 mM lAOx, 10 pM POD as catalyst, 10 pM riboflavin and 100 mM Tris-HCI pH 7.4 in 1 mL final volume. The remaining conditions are shown in FIG.2. Values are the mean of 5 replicates ± S.E. Group letters indicate significant differences (P value <0.01) in unidirectional ANOVA test and Tukey's post hoc test.

[0028] FIG. 6 shows NO2- production catalyzed by the xanthine oxidoreductase (XDH) enzyme from lAOx as substrate in normoxia, in the presence or not of the substrate xanthine or the reduced nicotinamide dinucleotide reductor (NADH), as indicated. Reactions were performed in airtight glass vials containing 2 mM lAOx, 10 pM XDH enzyme, 1 mM NADH and 100 mM Tris-HCI pH 7.4 in 1 mL. All other conditions as indicated in FIG. 2. Values are the mean of 4 replicates ± S.E. Group letters indicate significant differences (P value <0.01) in unidirectional ANOVA test and Tukey's post hoc test.

[0029] FIG. 7 shows the production of NO catalyzed by horseradish peroxidase (POD) using different aldoximes as substrate, as indicated in the figure caption (1 H-lndol-3- acetaldoxime, lAOx; 1 H-lndol-3-carboxaldoxime, IMOx; 2-(2,3-dihydro-1 H-inden-3-yl)- acetaldoxime, IDOx; benzaldoxime, BenzOx; 2-(4-hydroxyphenyl)-acetaldoxime, PhenOx), or various metabolites of the IAA (indole-3-acetic acid) biosynthesis pathway from Trp (Tryptophan, Trp; N-hydroxyl-tryptamine, NHT; IAA) by chemiluminescence. The substrate O-Methyl-1 H-lndol-3-acetaldoxime (lAOx-Met) as a control with the aldoxime group blocked. Reactions were performed in airtight glass vials containing 2 mM aldoxime or NHT or L-Arg or IAA, 10 pM hemoprotein as catalyst, 10 pM flavin and 100 mM Tris- HCI pH 7.4 in 1 mL. The remaining conditions are indicated in FIG. 2. Values are the mean of 5 replicates ± S.E. Asterisks indicate significant differences (p-value <0.001) in unidirectional ANOVA test and Dunnett's, post hoc test using lAOx-Met + POD + RbF as control.

[0030] FIG. 8 shows the determination by chemiluminescence of NO, nitrite and nitrate from lAOx as a result of the catalysis of different types of hemoproteins (rice Hb1 Lbl I , and horseradish peroxidase -POD) in the presence of the flavins flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN) and riboflavin (RbF). Reactions were performed in sealed glass vials containing 2 mM oxime, 10 pM hemoprotein as catalyst, 10 pM of any of the flavins, and 100 mM Tris-HCI pH 7.4 in 1 mL. The other conditions are indicated in Figure 5. The NO content was analyzed in a NO Analyzer (Serinus 40, Ecotech). Values are the mean of 5 replicates ± S.E. Group letters indicate significant differences (P value <0.01) in Tukey's one-way ANOVA and post hoc within each hemoprotein; asterisks indicate significant differences (p-value <0.001) in Dunnett's one-way ANOVA and post hoc test using lAOx + POD as control.

[0031] FIG. 9 is a visual Example of the effect of studied aldoximes on Medicago truncatula seedlings at day 4thand 8th.

[0032] FIG. 10 shows the effect of BenzOx, ButOx, and PhenOx , and of IDOx and IMOx compared to the control (grown with nitrate) on the number of secondary roots of M. truncatula seedlings grown for 12 days under 1 mM NOT. Values are the mean of 15-20 replicates ± S.E. Letters indicate significant differences at a = 0.05 between treatments using Student-Newman-Keuls tests.

[0033] FIG. 11 shows NO detected by chemiluminescence as catalyzed by mouse iNOS using L- Arg or lAOx as substrate after 10 min. Inset shows NO2' and NOT accumulated from the same reaction after 2 h. oblique lined and open bars are the sum of the average measures of NO2' and NOT derived. Dotted bars are direct NO measures by chemiluminescence. Data are mean ± S.E.M. n=5, group letters denote P < 0.01 statistical differences in unpaired t-test within each treatment; * denote statistical differences (P < 0.001) in unpaired t-test between treatments. FIG. 12 shows the effect of IDOx on the whole plant biomass at day 4 on shoot (A), root (B) and total biomass (C) of 14 days-old M. truncatula seedlings. Plants were grown under 1 mM nitrate, 1 mM NH4+or 1 mM NH4++ 100 pM IDOx, as indicated. Data represent means ± SE values (n=4-5). Different letters denote statistically significant differences at P < 0.05.

[0034] FIG. 13 shows the effect of IDOx, on the primary root length of M. truncatula seedlings on the growth period at day 4 (A) and 14 (B), and on the number of secondary roots (C) at day 14 (D). Data represent means ± SE values (n=4-5). Different letters denote statistically significant differences at P < 0.05.

[0035] FIG. 14 shows representative images of the phenotypic changes induced in M. truncatula seedlings growth under 1 mM nitrate, 1 mM NH4+or 1 mM NH4++ 100 M IDOx at day 4 (A) and 14 (B) by different Scale bar=1 cm.

[0036] FIG. 15 shows the rresults of the effect of the compounds 240x on the growth of S. aureus, in strains 15981 and V329. The photographs show tenfold dilutions of bacterial cultures grown in the presence of the indicated concentrations of the 240x compound.

[0037] FIG. 16 shows the rresults of the effect of the compounds on the biofilm formation of S. aureus strain 15981 (polysaccharide nature) at different concentrations. Graphs represent biofilm quantification from three independent experiments.

[0038] FIG. 17 shows the rresults of the effect of the compounds on the biofilm formation of S. aureus strain V329 (protein nature) at different concentrations. Graphs represent biofilm quantification from three independent experiments.

[0039] FIG. 18 shows the rresults of the effect of the compounds on the biofilm formation of S. Enteritidis strain 3934 at different concentrations.

[0040] Detailed description of the invention

[0041] Definitions

[0042] All terms as used herein in this application, unless otherwise stated, shall be understood in their ordinary meaning as known in the art. Other more specific definitions for certain terms as used in the present application are as set forth below and are intended to apply throughout the description and claims.

[0043] RECTIFIED SHEET (RULE 91 ) ISA / EP The term “pharmaceutically acceptable salts” used herein encompasses any salt formed from pharmaceutically acceptable non-toxic acids including inorganic or organic acids. There is no limitation regarding the salts, except that if used for therapeutic purposes, they must be pharmaceutically acceptable.

[0044] The term “acceptable salt for plants” used herein encompasses any salt formed from acceptable non-toxic acids including inorganic or organic acids. There is no limitation regarding the salts, except that if used for plants, they must be appropriate for such use.

[0045] The preparation of pharmaceutically acceptable salts or acceptable salt for plants of the aldoximes of the present invention can be carried out by methods known in the art. For instance, they can be prepared from the parent compound, which contains a basic moiety, by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the base form of these compounds with a stoichiometric amount of the appropriate pharmaceutically acceptable acid in water or in an organic solvent or in a mixture of them.

[0046] The compounds of the invention may be in crystalline form either as free solvation compounds or as solvates (e.g., hydrates) and it is intended that both forms are within the scope of the present invention. Methods of solvation are generally known within the art.

[0047] The term “solvate” refers to a molecular complex comprising any of the aldoximes or any of their salts or isomers according to the present invention, and a stoichiometric or non- stoichiometric amount of one or more solvent molecules bound by non-covalent intermolecular forces. When the one or more solvent molecules forming part of the molecular complex is water, the solvate is a hydrate. They can be prepared by methods known in the art, such as crystallization in an appropriate solvent.

[0048] The term “isomers” means compounds having the same number and kind of atoms, and hence the same molecular weight but differing with respect to the arrangement or configuration of the atoms in space.

[0049] The expression "therapeutically effective amount" as used herein in the context of a pharmaceutical composition comprising an aldoxime of the present invention, refers to the amount of aldoxime that, when administered, is sufficient to function in vivo as NO producers and NO donors, through a NO release process consisting of a redox reaction catalyzed enzymatically by peroxidases in vivo either at neutral pH or at physiological pH; and therefore it is an amount that prevent development of, or alleviate to some extent, one or more of the symptoms of the disease which is addressed, which are diseases sensible to the action of the NO. The particular dose of compound administered according to this invention will of course be determined by the particular circumstances surrounding the case, including the compound administered, the route of administration, the particular condition being treated, and the similar considerations.

[0050] The expression "pharmaceutically acceptable excipients or carriers" refers to pharmaceutically acceptable materials, compositions or vehicles. Each component must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the immediate release formulation. It must also be suitable for use in contact with the tissue or organ of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0051] The expression "effective amount of an aldoxime capable of releasing nitric oxide (NO) in vivo " as used herein in the context of a composition for plants refers to the amount of an aldoxime that, when administered, alone or in combination with other active compounds, is sufficient for stimulating lateral root development and potentiating the root structure of plants.

[0052] “Physiological pH in mammals”, in particular, in humans, refers to the normal range of pH values found in mammals, particularly in the human body. It represents the normal range of pH values that are necessary for proper functioning of various biological processes. The pH of the body is an essential component of maintaining homeostasis, and deviations from the normal physiological pH range can have detrimental effects on health. pH is a measure of acidity or alkalinity and is determined by the concentration of hydrogen ions (H+) in a solution. For instance, in mammals, in particular, in the human body, different organs, tissues, and fluids have specific pH ranges that are necessary for proper functioning. The overall physiological pH range typically varies from 7.3-7.5, in particular, 7.35 to 7.45 in blood. This range is slightly alkaline or basic, indicating that the internal environment of the body is more alkaline than neutral.

[0053] “Neutral pH” means a pH around 7.

[0054] “Physiological pH in plants” is broader than in mammals as generally is considered to be a pH in the range from 5 to 8.

[0055] “Abiotic stress” refers to the negative impact of non-living factors on living organisms in a specific environment. The stresses include drought, salinity, low or high temperatures, and other environmental extremes. Abiotic stresses, especially drought and hipersalinity, are the primary causes of crop loss worldwide.

[0056] “Biotic stress” means stress that occurs as a result of damage done to an organism by other living organisms, such as bacteria, viruses, fungi, parasites, beneficial and harmful insects, weeds, and cultivated or native plants.

[0057] As mentioned above, it is part of the present invention an aldoxime able of releasing nitric oxide (NO) for use in the treatment of a condition or a disease which is sensible to the action of the NO, wherein the treatment comprises administering the aldoxime to a subject and releasing NO from the aldoxime through an in vivo enzymatically catalyzed redox reaction either at neutral or at physiological pH. This aspect of the present invention can alternatively be formulated as the use of an aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof as defined above for the manufacture of a medicament for the treatment of a condition or a disease which is sensitive to the action of the NO.

[0058] This aspect can alternatively be formulated as a method for the treatment of a condition or a disease which is sensitive to the action of the NO, the method comprising administering a therapeutically effective amount of an aldoxime, or a pharmaceutically acceptable salt thereof or an isomer thereof as defined above in combination with pharmaceutically acceptable excipients or carriers to a subject in need thereof.

[0059] In the context of the present invention, and as used herein when referring to aldoximes, it refers to the aldoximes, or their pharmaceutically acceptable salts thereof or their isomers thereof, i.e., it is used in generic form to encompass any of them.

[0060] Thus, the treatment is to be conducted in a subject, which can be a mammal, including a human. By administering the aldoximes to the subject there is an increase in the oxime level in the body that is oxidized in vivo and release NO, which produces many beneficial biological effects. The effect of aldoximes is concentration dependent.

[0061] In a particular embodiment, the aldoxime able of releasing nitric oxide (NO) is for use in the treatment of a condition or a disease which is sensible to the action of the NO, wherein the treatment comprises administering the aldoxime to a subject and releasing NO from the aldoxime through an in vivo enzymatically catalyzed redox reaction at physiological pH.

[0062] In another particular embodiment, the aldoxime for use according to the present invention is that which is able to of releasing nitric oxide (NO) through an enzymatically catalyzed redox reaction in vivo either at neutral or at physiological pH, wherein the reaction is catalyzed by a peroxidase or a nitric oxide synthase.

[0063] In another particular embodiment, the aldoxime is for use in the treatment of a condition or disease which is selected from the group consisting of: vasodilation, hypertension, a cardiovascular disease, an acute episode of cardiovascular disease, a cancerous tumor, an infectious disease, inflammatory condition, in particular, as a signaling agent of the antioxidant response, pathogen infection, and in the prevention of oxidative burst that occurs in situations of ischemia-reperfusion in open heart surgery.

[0064] In another particular embodiment, the aldoxime is for use according to the present invention, wherein the acute episode of cardiovascular disease is a heart attack.

[0065] In another particular embodiment, the aldoxime is for use according to the present invention, wherein the cardiovascular disease is mediated by the vasodilator effect of the NO or by the anti-inflammatory effect of the NO.

[0066] In another particular embodiment, the aldoxime for use according to the present invention is that where the cancerous tumor or the inflammatory condition are mediated by the antiinflammatory effect of the NO.

[0067] In another particular embodiment, the aldoxime for use according to the present invention is that where the cardiovascular disease, the cancerous tumor or the inflammatory condition are mediated by the anti-inflammatory effect of the NO.

[0068] In another particular embodiment, the aldoxime for use according to the present invention is that which is used to treat breast cancer. The examples (table 1) illustrate the toxicity of the aldoxime lAOx in the growth of breast cancer cells MCF-7.

[0069] In another particular embodiment, the aldoxime for use according to the present invention is that which is used against a pathogen infection caused by bacteria. The bacteria can be gram-positive or gram-negative bacteria.Jn another particular embodiment, the aldoxime for use as defined above, is that where the aldoxime is for use in combination therapy with one or more known antibacterial agents.

[0070] In another particular embodiment, the aldoxime for use according to the present invention is that which is selected from the group consisting of 1 H-lndol-3-acetaldoxime (lAOx), 2- (2,3-dihidro-1 H-inden-3-il)-acetaldoxime (IDOx), 1 H-lndol-3-carboxaldoxime (IMOx), 2- methyl-Butylaldoxime (ButOx), 2-(4-hidroxiphenyl)-acetaldoxime (PhenOx), benzaldoxime (BenzOx), naphtyl-1 -acetaldoxime (NaftOx), 2,4-chlorophenoxyacetaldoxime (2,4-CIOx) and O-methyl-1 H-indole-3-acetaldoxime (lAO-Met).

[0071] In another particular embodiment, the aldoxime for use according to the present invention is that which is selected from the group consisting of: 2-(2, 3-di hidro- 1 H-inden-3-il) acetaldoxime (IDOx), Indolacetaldoxime (lAOx), O-Methyl-1 H-lndol-3-acetaldoxime (lAOX-met), lndol-3-carboxaldoxime (IMOx), 4-hidroxiphenilaldoxime (PhenOx), Benzilaldoxime (BenzOx), and Butylaldoxime (ButOx).

[0072] The aldoximes are generally administered in patients by intravenous route. It can be injected for instance, in a saline solution with a small amount of tween.

[0073] In a particular embodiment, the aldoxime for use according to the present invention is that which is indole-3-acetaldoxime (lAOx). The methylated form of this aldoxime also has phenotypic effect. This is attributed to the demethylation of the molecule once internalized in the plant cell.

[0074] Flavins and peroxidases (POD) both catalyze the production of NO from aldoximes and modulate the rate of NO production, and thus the biological effect. Also, the Nitric Oxide Synthase enzyme catalyze the reaction with a little lower efficiency. The examples of the present invention illustrate that a mixture of flavin, POD and aldoxime affected the growth of cancer cells (MCF-7) in vitro to a greater extent than non-cancerous control cells. Thus, in another particular embodiment, an aldoxime for use according to the present invention is that where the compound that catalyze the in vivo release of nitric oxide (NO) from the aldoximes is selected from the group consisting of a peroxidase, a flavin, a hemoglobin, a nitric oxide synthase, and a mixture thereof. In another particular embodiment, the in vivo use of an aldoxime according to the present invention is that where the flavin is selected from the group consisting of FAD, FMN, and Riboflavin. In another particular embodiment, an aldoxime for use according to the present invention is that where the compound that catalyze the in vivo release of nitric oxide (NO) from the aldoximes is a mixture of flavin and hemoglobin.

[0075] In another particular embodiment, the aldoxime for use according to the present invention is that which is able to of releasing nitric oxide (NO) through an enzymatically catalyzed redox reaction in vivo either at neutral or at physiological pH, wherein the reaction is catalyzed by a nitric oxide synthase.

[0076] The oximes of the present invention due to their ability of releasing nitric oxide can also provide a beneficial effect on sport performance. Thus, the use of an aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof able of releasing nitric oxide (NO) to improve the muscle contractility capacity, ventilatory efficiency, and sports performance is also part of the invention,

[0077] An in vivo use of an aldoxime able of releasing nitric oxide (NO) in vivo, for biological applications which are sensible to the action of the NO in plants, wherein the biological application is selected from the group consisting of plant rooting agent, plant structure modifier, stimulators of the tolerance to abiotic or biotic stress, plants protection from stress induced by ammonium nutrition as an exclusive source of nitrogen, plants protector from hydric stress, and as germination inducer.

[0078] In a particular embodiment of the in vivo use of the aldoxime the biological application is selected from the group consisting of plant rooting agent and plant structure modifier. Acting as plant rooting agent or plant structure modifier include for instance acting as lateral root development stimulator, root surface increasing agent, and aerial structure and bearing modulator. Thus, it is possible to obtain customized root structures, increasing the root surface, and even modulating the aerial structure and bearing. The effect of the aldoximes is dependent on the concentration of aldoxime.

[0079] In a particular embodiment, the in vivo use of the aldoxime is that where the biological application is selected from the group consisting of plant rooting agent and plant structure modifier.

[0080] In a particular embodiment, the in vivo use of the aldoxime is that where the biological application is as plants protector from hydric stress, wherein the hydric stress is for example drought,

[0081] In another particular embodiment, the in vivo use of an aldoxime able of releasing nitric oxide (NO) in vivo, is that where the use comprises administering the aldoxime to a plant and releasing NO from the aldoxime through an in vivo enzymatically catalyzed redox reaction at a pH from 5 to 8. In a more particular embodiment, the pH is in the range from 5 to 7.5. In another particular embodiment, the pH is in the range 5.5. to 7.5.

[0082] In a particular embodiment, the in vivo use of an aldoxime according to the present invention as a plant structure modifier is due to its effect on the formation of leaf primordia and on the structure of the aerial part of the plant.

[0083] Generally, plants are usually grown with ammonium nitrate or nitrate simply as nitrogen fertilizers. The urea is also sometimes used as it is cheaper But, the exclusive use of ammonium is more environmentally friendly since ammonium is not as easily washed out of the soil as nitrate, and therefore ammonium nutrition does not contribute to surface water pollution. As a way of example the potent induction of root growth shown by IDOx treatment is remarkable, in particular, the I DOx-treated plant grows more at day 14 than the control (under ammonium nutrition conditions). As it is illustrated in the examples, IDOx induces a higher plant growth in ammonium both in terms of root length and in the number of secondary roots. In addition to stimulating root growth, it induces the growth in biomass of the aerial part and therefore of agricultural production, if the economically important part is the aerial part, as is often the case. This may also be important in drought tolerance. Accordingly, in a particular embodiment, the in vivo use of an aldoxime according to the present invention is as a plant protector from stress induced by ammonium nutrition as an exclusive source of nitrogen. In another particular embodiment, the in vivo use of the aldoxime IDOx is as a plant protector from stress induced by ammonium nutrition as an exclusive source of nitrogen.

[0084] In another particular embodiment, the in vivo use according to the present invention is that where the aldoxime is selected from the group consisting of 1H-lndol-3-acetaldoxime (lAOx), 2-(2,3-dihidro-1 H-inden-3-il)-acetaldoxime (IDOx), 1 H-lndol-3-carboxaldoxime (IMOx), 2-methyl-Butylaldoxime (ButOx), 2-(4-hidroxiphenyl)-acetaldoxime (PhenOx), benzaldoxime (BenzOx), naphtyl-1 -acetaldoxime (NaftOx), 2,4- chlorophenoxyacetaldoxime (2,4-CIOx) and O-methyl-1 H-indole-3-acetaldoxime (IAO- Met).

[0085] In another particular embodiment, the in vivo use according to the present invention is that where the aldoxime is selected from the group consisting of 2-(2,3-dihidro-1 H-inden-3- il)acetaldoxime (IDOx), indolacetaldoxime (lAOx), O-methyl-1 H-lndol-3-acetaldoxime (lAox-met), lndol-3-carboxaldoxime (IMOx), 4-hidroxiphenylaldoxime (PhenOx), benzilaldoxime (BenzOx), and butylaldoxime (ButOx).

[0086] The aldoximes are stable in the absence of catalyst. Although some of the catalysts (POD, XDH, flavins) used are present in biological tissues of both plants and animals, in general, there is the possibility that the use of aldoximes may be accompanied in the formulation of the catalyst depending on the modulation of the biological signal to be obtained. Thus, in a particular embodiment, the in vivo use of an aldoxime according to the present invention is that where the aldoxime is used together with a compound that catalyzes the in vivo release of nitric oxide (NO) from the aldoximes.

[0087] The catalysts can be peroxidases such as horseradish peroxidase; hemoglobin’ such as rice hemoglobin 1 or cowpea leghemoglobin II; flavohemoglobins such as nitric oxide synthase; flavoproteins such as YUCCA-type proteins in plants or animal flavinmonooxygenase; as well as free flavins including riboflavin, flavin adenine mononucleotide and flavin adenine dinucleotide. In another particular embodiment, the in vivo use of an aldoxime according to the present invention is that where the compound that catalyze the in vivo release of nitric oxide (NO) from the aldoximes is selected from the group consisting of a peroxidase, a flavin, and a hemoglobin.

[0088] In another particular embodiment, the in vivo use of an aldoxime according to the present invention is that where the flavin is selected from the group consisting of FAD, FMN, and Riboflavin.

[0089] In plants, aldoximes can be added for instance as follows: 100 - 200 pM of the aldoxime can be diluted in an appropriate solvent such as DMSO and added to each of either solid (agar containing) or liquid media.

[0090] All the aldoximes exemplified in the present invention have been obtained by chemical synthesis as illustrated in the examples.

[0091] The aldoximes may be combined with hormones such as indoleacetic acid (IAA), due to its capacity of producing NO.

[0092] It is also part of the invention the compound of formula 2-(2, 3-di hidro-1 H-inden-3-il) acetaldoxime (IDOx) perse.

[0093] For the therapeutic uses, the aldoximes may be administered in the form of pharmaceutical composition. The pharmaceutical composition comprises a therapeutically effective amount of an aldoxime capable of releasing nitric oxide (NO) in in vivo systems, together with pharmaceutically acceptable excipients or carriers.

[0094] Pharmaceutically acceptable carriers include at least one of the following agents: a solvent such as water a buffer such as phosphate buffer saline, or an emulsifier.

[0095] The pharmaceutical composition may be formulated into a dosage form such as injection (e.g., sterile aqueous solutions, dispersions, etc.).

[0096] The aldoximes may also form part of a composition for plants comprising an effective amount of an aldoxime capable of releasing nitric oxide (NO) in vivo, together with appropriate carriers, wherein the effective amount is for stimulating fruit and root development and for modifying plant structure. Throughout the description and claims the word "comprise" and variations of the word, are not intended to exclude other technical features, additives, components, or steps. Furthermore, the word “comprise” encompasses the case of “consisting of”. Additional objects, advantages and features of the invention will become apparent to those skilled in the art upon examination of the description or may be learned by practice of the invention. The following examples and drawings are provided by way of illustration, and they are not intended to be limiting of the present invention. Reference signs related to drawings and placed in parentheses in a claim, are solely for attempting to increase the intelligibility of the claim and shall not be construed as limiting the scope of the claim. Furthermore, the present invention covers all possible combinations of particular and preferred embodiments described herein.

[0097] Examples

[0098] Example 1 : Preparation of Methyl-(2,3-dihydro-1 / 7-inden-3-yl)-ylidenacetate (IDOx precursor)

[0099] In a three-necked round-bottomed flask, a suspension of Zn (2610 mg, ca. 40.0 mmol) and 1-indanone (3100 mg, 23.5 mmol) in anhydrous THF (90 mL) was stirred at 60 °C for 5 minutes. Then, a solution of methyl-2-bromoacetate (3.300 mL, 35.0 mmol) in anhydrous THF (50 mL) was added dropwise during 20 min. The resulting solution was stirred at 65 °C until the initial product was reacted (TLC, SiCh, DCM; ca. 1 h) and then it was allowed to cool. HCI (1 N, 100 mL) was added, the suspension was stirred for 5-10 min and extracted with ethyl acetate (3 x 100 mL). The combined organic phases were washed with brine (100 mL) and dried with magnesium sulphate. The solvent was then removed under reduced pressure providing a yellowish liquid that was purified by flash chromatography (SiO2, DCM / hexanes 2:1) to yield methyl-(2,3-dihydro-1 / 7-inden-3-yl)- ylidenacetate (342 mg, 48 %) as a mixture of two isomers characterized by NMR.

[0100] Major isomer, 75 %.1H NMR (CDCh, ppm):7.60 (d, J=7.76, 1 H, H7), 7.39-7.35 (m, 2H), 7.28-7.19 (m, 1 H) 6.32 (dd o t, J= 2.62, 1 H, H1), 3.77 (s, 3H, CH3), 3.33-3.28 (m, 2H, H2’), 3.09 (d, J= 6.68, 1 H, H3’), 3.07 (d, J= 5.84, 1 H, H3’),

[0101] 13C NMR (CDCI3): 163.38 (C1), 149.67 (C3a’),140.02(CT), 136.73 (C7a’), 131.03 (C6’), 126.89, 125.76, 121.76 (C7’), 107.24 (C2), 51.12 (CH3), 31.30 (C3’), 30.67 (C2’)

[0102] Minor isomer, 25 %.1H NMR (CDCh, ppm): 7.46 (dt, J=7.38, 0.87Hz, 1 H), 7.39-7.35 (m, 1 H), 7.34-7.28 (m, 1 H), 7.28-7.19 (m, 1 H), 6.44 (dd o t, J= 1.65, 1 H,), 3.72 (s, 3H, CH3), 3.62 (dd, J= 3.06, 1.58, 1 H, H3’), 3.33-3.28 (m, 2H, H2’), 3.40-3.37(m, 2H, H3’),13C NMR (CDCh): 168.06 (C1), 144.51 (CT), 144.16 (C3a’), 136.73 (C7a’), 131.91 (C2), 126.30, 124.99, 123.91 , 119.22, 52.11 (CH3), 38.04 (C3’), 33.96 (C2’)

[0103] Example 2: Preparation of methyl 2-(2,3-dihydro-1 / 7-inden-3-yl)acetate (I DOX precursor)

[0104] A solution of methyl 2-(2,3-dihydro-1 / 7-inden-3-yl)-ylidenacetate (1000 mg. 5.32 mmol), Pd / C (10%. 56 mg, 0.053 mol Pd) in methanol (20 mL) was charged in a Fisher-Porter bottle. The bottle was pressurized with H2 (20 psi) and the suspension was stirred at room temperature during 24 h. The pressure was then released, and the suspension was filtered through a pad of celite. The solvent was removed by rotary elimination yielding methyl 2-(2,3-dihydro-1 / 7-inden-3-yl)-acetate (960 mg, 95%) as a yellowish oil.1H NMR (CDCh, ppm): 7.25-7.20 (m, 1 H), 7.19-7.15 (m, 3H), 3.73 (s, 3H, CH3), 3.64-3.53 (m, 1 H, HT), 2.99-2.83 (m, 2H, H3’), 2.45 (dd, J= 15.50, 9.19, H2), 2.45 (dd, J= 15.50, 9.19, H2), 2.45-2.43 (m, 1 H, H2’), 1.79-1.70 (m, 1 H, H2’).13C NMR (CDCh, ppm): 173.39 (C1), 145.80 (C7a’), 144.03 (C3a’), 123.94, 123.45, 124.76, 123.56, 51.74 (CH3), 41.48 (CT), 39.85 (C2), 32.55 (C2’), 31.32 (C3’)

[0105] Example 3: Preparation of 2-(2,3-dihydro-1 / 7-inden-3-yl)acetaldehyde (I DOX precursor)

[0106] A stirred solution of methyl 2-(2,3-dihydro-1 / 7-inden-3-yl)-acetate (912 mg, 4.80 mmol) in anhydrous toluene (50 mL) was cooled to -80 °C. Then, a DIBAL-H solution (1.2M, 5.2 mL, 6.2 mmol, 1.2 eq) was added dropwise keeping the reaction temperature below - 75 °C. The resulting solution was stirred at -80 °C for 30 min. and then methanol (10 mL) was carefully added. The solution was vigorously stirred and allowed to reach room temperature. HCI (1 N, 130 mL) was added, and the solution was stirred for additional 10 min. The organic phase was separated, and the aqueous layer was extracted with toluene (2x 20 mL). The combined organic phases were washed with Na2CC (20 mL) and dried with magnesium sulfate. The solvent was evaporated under vacuum yielding to obtain a liquid that was purified by flash chromatography (SiCh, hexanes / ethyl acetate 4:1) to yield 2-(2,3-dihydro-1 / 7-inden-3-yl)-acetaldehyde (377 mg, 49 %.) as a colorless oil.1H NMR (CDCh, ppm): 9.90 (t, J= 2.08, 1 H, H1), 7.23 (d, J= 3.67, 1 H, H4’), 7.20-7.30 (m, 3H), 3.73-3.63 (m, 1 H, HT), 3.00-2.83 (m, 3H, H2, H3’), 2.63 (ddd, J= 17.04, 8.06, 1.92, H2), 2.49-2.38 (m, 1 H, H2’), 2.49-2.38 (m, 1 H, H2’), 1.77-1.66 (m, 1 H, H2’).13C NMR (CDCh, ppm): 201.03 (C1), 145.58 (C7a’), 143.93(C3a’), 126.98 (C5’), 126.55, 124.81 (C4’), 123.66, 123.52, 49.59 (C2), 39.14 (CT), 32.64 (C2’), 31.50 (C3’).

[0107] Example 4: Preparation of 2-(2,3-dihydro-1 / 7-inden-3-yl)acetaldoxime (IDOx) To a stirred solution of 2-(2,3-dihydro-1 / 7-inden-3-yl)-acetaldehyde (373 mg, 2.33 mmol) in toluene / H2O (1 :1 , 100 mL) aqueous hydroxylamine was added (50% aq. 856 pL, 14.0 mmol, 6 eq). The resulting solution was vigorously stirred at room temperature for 20 hours and then the organic phase was separated. The aqueous layer was extracted with toluene (50 mL). The combined organic phases were dried with magnesium sulfate. The solvent was evaporated under vacuum providing a solid that was purified by flash chromatography (SiO2, hexanes / ethyl acetate / methanol 4:1 :0.5) to yield 2-(2,3-dihydro- 1 / 7-inden-3-yl)-acetaldoxime (220 mg, 55%)

[0108] Z-isomer, 39 %.1H NMR (CDCh, ppm): 7.63 (1 H, OH, b,), 7.27-7.20 (2H, m, H4in, H7in), 7.20-7.12 (2H, m, H5in, H6in), 6.83 (1 H, t, J= 5.5, N=CH), 3.45-3.33 (1 H, m, H1in), 3.02- 2.80 (3H, m, H2 (2.90), H3in), 2.61 (2.67-2.55 1 H, m, H2), 2.34 (2.40-2.27 1 H, m, H2in), 1.76 (1.83-1.69 1 H, m, H2in).13C NMR (CDCh, ppm): 151.5 (C1 , N=CH), 145.9 (C7ain), 144.1 (C3ain), 126.9 126.5, (C5iny C6in) 124.8 (C4in), 123.6 (C7in), 42.1 (C1in), 32.2 (C3 in), 31.4 (C2 in), 30.0 (C2).

[0109] E-isomer, 61 %.1H NMR (CDCh, ppm): 7.49 (1 H, t, J= 6.2, N=CH), 7.25-7.21 (m, 2H, H4’, H5’), 7.19-7.15 (m, 1 H, H6’, H7’), 3.42-3.36 (m, 1 H, HT), 3.00-2.83 (m, 3H, H2 (2.70), H3’), 2.70 (m, 1 H, H2), 2.42 (m, 1 H, H2’), 1.83-1.69 (1 H, m, H2’).13C NMR (CDCh, ppm): 151.22 (C1), 145.75 (C7a’), 144.10 (C3a’), 126.87, 126.44, 124.78 (C4’), 123.71 (05’), 42.64 (CT), 34.8 (C2), 31.76 (C2’), 31.83 (C3’). [M+H+]: 176.1804 (calc, for CH HI4NO+: 176.1075).

[0110] Example 5: Preparation of 1 H-lndole-3-carbaldehyde oxime (IMOx)

[0111] To a stirred solution of 1 / 7-indole-3-carboxaldehyde (1450 mg, 10.0 mmol) in ethanol (50 mL), aqueous hydroxylamine (50%, 3.7 mL, 60 mmol) was added. The resulting solution was refluxed for 18 h and then allowed to cool. The solvent was removed by rotary evaporation yielding indol-3-carboxaldoxime (1470 mg, 92%) as an orange solid.

[0112] Z-lsomer, 78 %.1H NMR (DMSO-d6): 11.55 (b,1 H, NH), 11.16 (s,1 H, OH), 8.21 (1 H, d, J= 2.68, H2m) 7.85 (d, J= 7.80, 1 H, H4in),7.78 (s, 1 H, =CH), , 7.44 (ddd, J= 8.08, 1.05, 0.92, 1 H, H7in), 7.17 (ddd, J= 6.96, 6.84, 1.24, H6in), 7.11 (ddd, J= 8.08, 6.97, 1.18, H5in).13C NMR (DMSO-d6): 138.34 (=CH), 134.89 (C7a), 130.42 (C2in), 126.20 (C3ain), 121.83 (C6in) 118.15 (C4in), 111.73 (C7in), 119.86 (C5in), 106.30 (C3in).

[0113] E-isomer, 22%.1H NMR (DMSO-d6): 11.37 (b,1 H, NH), 10.47 (s,1 H, OH), 8.26 (s, 1 H, =CH), 7.97 (d, J= 7.84, 1 H, H4in7.623 (1 H, d, J= 2.64, H2m), 7.41 (d, J= 7.96, 1.12, 1.02, 1 H, H7in, 7.17 (ddd, J= 6.96, 6.84, 1.24, H6in), 7.11 (ddd, J= 8.08, 6.97, 1.18, H5in)13C NMR (DMSO-d6): 144.55 (=CH), 136.85 (C7ain), 128.4 (C2in), 124.21 (C3ain), 122.22 (C6in), 121. 42 (C4in), 111.73 (C7in), 119.98 (C5in), 109.59 (C3in). [M+H+]: 161.0725 (calc, for C9H9N2O+: 161.0715).

[0114] Example 6: Preparation of Benzaldoxime (BenzOx)

[0115] To a solution of benzaldehyde (1015 pL, 10.0 mmol) in toluene / H2O (1 :1 , 100 mL) aqueous hydroxylamine (50% aq. 3.7 mL, 60.0 mmol) was added. The solution was vigorously stirred at room temperature for 20 hours. Then the organic phase was separated, and the aqueous layer was extracted with toluene (50 mL). The combined organic phases were dried with magnesium sulfate and the solvent was evaporated under vacuum yielding benzaldoxime (1125 mg, 93%) as an oily syrup.

[0116] E-isomer, 93%.1H NMR (CDCh, ppm): 8.17 (1 H, t, J= 6.4, HON=CH), 7.70 (1 H, b, OH), 7.61-7.55 (2H, m, H2, H6), 7.42-7.37 (3H, m, H3, H4, H5).13C NMR (CDCh, ppm): 150.6 (ON=C), 132.2(C1), 130.2 (C4), 128.9 (C3, C5), 127.2 (C2, C6).

[0117] Z-isomer, 7%.1H NMR (CDCh, ppm): 8.14 (1 H, t, J= 6.4, HON=CH), 7.97-7.92 (2H, m, H2, H6), 7.70 (1 H, b, OH), 7.46-7.42 (3H, m, H3, H4, H5).13C NMR (CDCh, ppm): 147.3 (ON=C), 132.2 (C1), 131.1 (C4), 130.3 (C3, C5), 128.7 (C2, C6). [M+H+]: 122.0611 (calc. for C7H8NO+: 122.0606)

[0118] Example 7: Preparation of lndole-3-acetaldoxime (lAOx) lndole-3-acetaldoxime and its precursor, indole-3-acetaldehyde, were both obtained as described previously (Buezo et al.” lAOx induces the SUR phenotype and differential signalling from IAA under different types of nitrogen nutrition in Medicago truncatula roots”, Plant Science, 2019, 110176).

[0119] Example 8: Preparation of indole-3-acetaldehyde, O-methyl oxime (lAOx-Met) lndole-3-acetaldehyde was obtained as described (Buezo et al., 2019). To a stirred solution of sodium carbonate (505 mg, 4.81 mmol) and O-methylhydroxylamine (400 mg, 4.81 mmol) in H2O (30 mL) a solution of 1 / 7-1 ndole-3-acetaldehyde (153 mg, 0.96 mmol) in ethanol (40 mL) was added. The resulting solution was stirred at room temperature during 18 h and then, concentrated by rotary distillation. A mixture of toluene: H2O (1 :1 , 100 mL) was added, the organic layer was separated, and the aqueous layer was extracted with toluene (2 x 25 mL). The combined organic phases were washed with brine (25 mL) and dried with magnesium sulfate. The solvent was removed by vacuum distillation to yield 1 / 7-1 ndole-3-acetaldehyde, O-methyloxime (150 mg, 80%) E-isomer, 64 %.1H NMR (CDCh, ppm): 8.02 (1 H, b, NH), 7.63 (1 H, dd, J= 7.9, 1.1 , H4in), 7.54 (1 H, t, J= 6.4, ON=C / 7), 7.37 (1 H, dt, J= 8.1 , 0.9, H7in), 7.22 (1 H, dd, J= 7.6, 1.5, 1.3, H6in) ,7.14 (tad, J= 7.5, 1.0, 0.9, H5in), 7.06 (1 H, b, H2in), 3.87 (3H, s, CH3), 3.66 (2H, dd, J= 6.4, 0.9, CH2).13C NMR (CDCh, ppm): 149.4 (C1), 136.5 (C7ain), 127.4 (C3ain), 122.5 (C2in), 122.3 (C6in), 119.8 (C5in), 119.0 (C4in), 111.3 (C7in), 111.0 (C3in), 61.5 (CH3), 26.1 (C2).

[0120] Z-isomer, 36 %.1H NMR (CDCh, ppm): 8.02 (1 H, b, NH), 7.58 (1 H, dd, J= 7.9, 1.2, H4in), 7.38 (1 H, dt, J= 8.1 , 1.1 , H7in), 7.22 (1 H, tad, J= 7.6, 1.5, 1.3, H6in), 7.14 (tdd, J= 7.5, 1.0, 0.9, H5in), 7.06 (1 H, b, H2in), 6.87 (1 H, t, J= 5.2, ON=CH), 3.98 (3H, s, CH3), 3.80 (2H, dd, J= 5.2, 0.9, CH2).13C NMR (CDCh, ppm): 150.4 (C1), 136.5 (C7ain), 127.4 (C3ain), 122.5 (C2in), 122.2 (C6in), 119.8 (C5in), 118.9 (C4in), 111.3 (C7in), 111.0 (C3in), 61.9 (CH3), 22.2 (C2). [M+H+]: 189.1039 (calc, for CH HI3N2O+: 189.1028).

[0121] Example 9: Preparation of methyl 2-(4-hydroxyphenyl)acetate (precursor or PhenOx)

[0122] A solution of 4-hydroxyphenyl-acetic acid (3000 mg, 19.7 mmol) and H2SO4 (96%, 3.0 mL) in methanol (150 mL) was refluxed during 2 h. The resulting solution was allowed to cool and concentrated by rotary evaporation. Ethyl acetate (100 mL) and NaHCO3sat. (100 mL) were added and the organic layer was collected. The aqueous layer was extracted with ethyl acetate (2 x 100 mL). The combined organic phases were washed with NaHCCh sat. (100 mL) and brine (100 mL) and dried with magnesium sulfate. The solvent was removed by vacuum evaporation yielding methyl- (4-hydroxyphenyl)-acetate (3060 mg, 94 %) as an oil.1H-NMR (CDCh, 6 ppm): 7.13 (2H, dt, J= 9.34, 2.47, H2Ph, H6Ph), 6.76 (2H, dt, J= 9.40, 2.50, H2Ph, H6Ph), 5.12 (1 H, s, O / 7), 3.70 (3H, s, OCW3), 3.56 (2H, s, CW2)

[0123] Example 10: Preparation of 2-(4-hydroxyphenyl)acetaldehvde (precursor or PhenOx)

[0124] A stirred solution of methyl 2-(4-hydroxyphenyl)-acetate (2000 mg, 12.2 mmol) in anhydrous toluene (120 mL) was cooled to -80 °C. Then, a DIBAL-H solution (1.2M, 51 mL, 61 mmol, 5.0 eq) was added dropwise keeping the reaction temperature below - 75 °C. The resulting solution was stirred at -80 °C for 30 min. and then methanol (10 mL) was carefully added. The solution was vigorously stirred and allowed to reach -10 °C and then, additional HCI (1 N, 200 mL) was added. The solution was allowed to reach room temperature and poured into a separation funnel. The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2x 100 mL). The combined organic layers were washed with NaHCCh (2 x 100 mL) and brine (2 x 75 mL) and dried with magnesium sulfate. The solvent was evaporated under vacuum to obtain a liquid that was purified by flash chromatography (SiO2, DCM / MeOH 20:1) to yield (4-hydroxyphenyl)- acetaldehyde (350 mg, 21 %).1H-NMR (CDCh, 6 ppm): 9.72 (1 H, t, J= 2.38, C / 70), 7.08 (2H, dt, J= 8.43, 2.56, H2Ph, H6Ph), 6.83 (2H, dt, J= 9.47, 2.48, H2Ph, H6Ph), 5.29 (1 H, s, 0 / 7), 3.62 (2H, d, J= 2.32, CH2).13C NMR (CDCh, ppm): 199.84 (CHO), 155.09 (C4Ph), 131.04 (C2phC6ph), 124.06 (C1ph), 115.05 (C3phC5ph), 49.85 (CH2).

[0125] Example 11: Preparation of 2-(4-hydroxyphenyl)acetaldoxime (PhenOx)

[0126] To a stirred solution of 2-(4-hydroxyphenyl)acetaldehyde (350 mg, 2.57 mmol) in ethanol (5 mL) aqueous hydroxylamine (50%, 952 pL, 15.4 mmol) was added. The solution was stirred at room temperature during 7 h and then the solvent was distilled off. The resulting oil was dissolved in DCM / H2O (1:1, 50 mL). The organic layer was removed, and the aqueous phase was extracted with DCM (25 mL). The combined organic phases were washed with brine (25 mL) and dried with magnesium sulfate. The solvent was removed by rotary evaporation providing a solid that was extracted with hot ethyl acetate (2 x 25 mL) to yield 2-(4-hydroxyphenyl)acetaldoxime (158 mg, 41%).

[0127] Z-isomer, 50%.1H-NMR (DMSO-d6, ppm): 10.93 (1H, s, N-OH), 9.23 (H, b, Ph-OH), 7.01 (2H, d, J=8.8, H2ph, H6ph), 6.69 (2H, dd, J=8.5, 2.1, H3ph, H5ph), , 6.74 (1 H, t, J= 5.3, N=C / 7), 3.48 (2H, d, J= 5.4, CH2).13C-NMR (DMSO-d6, ppm): 155.9 (C4ph), 149.1 (C1, N=CH), 129.6 (C2phC6ph), 127.3 (C1ph), 115.3 (C3phC5ph), 30.2 (C2)

[0128] E-isomer, 50%.1H-NMR (DMSO-d6, ppm): 10.47 (1 H,s, N-OH), 9.23 (H, b, Ph-OH), 7.35 (1 H, t, = 6.3, N=C / 7), 6.99 (2H, d, J=8.8, H2ph, H6ph), 6.69 (2H, dd, J=8.5, 2.1 , H3ph, H5ph), 3.32 (1 H, d, J= 6.9, CH2).13C-NMR (DMSO-d6, ppm): 155.8 (C4ph), 148.8 (C1, N=CH), 129.6 (C2phC6ph), 127.1 (C1ph), 115.3 (C3phC5ph), 34.4 (C2) [M+H+]: 152.0717 (calc, for C8HI0NO2+: 152.0712).

[0129] Example 12: Preparation of 2-methylbutanaldoxime (ButOx)

[0130] To a stirred solution of 2-methylbutyraldehyde (300 pL, 2.80 mmol) in ethanol (2 mL) aqueous hydroxylamine (50%, 1040 pL, 16.8 mmol) was added. The solution was stirred at room temperature during 16 h and then a mixture of DCM / H2O (1:1, 50 mL) was added. The organic layer was removed, and the aqueous phase was extracted with DCM (25 mL). The combined organic phases were washed with brine (25 mL) and dried with magnesium sulphate. The solvent was removed by rotary evaporation yielding 2- methylbutiraldoxime (170 mg, 60 %). E isomer, 74%.1H NMR (CDCh, ppm): 7.69 (1 H, s, OH), 7.30 (1H, d, J= 6.9, N=CH), 2.29 (1 H, sept,2CH), 1.56-1.34 (2H, m,3CH2), 1.07 (3H, d, CH3- ), 0.92 (3H, t, CH3-w).

[0131] 1H NMR (dmso-d6, ppm): 10.33 (1 H, s, OH), 7.18 (1 H, d, J= 6.4, N=CH), 2.19 (1 H, sept,2CH), 1.33 (2H, m,3CH2), 0.99 (3H, d, CH3- ), 0.84 (3H, t, CH3-w).

[0132] Z isomer, 26%.:1H NMR (CDCh, ppm): 7.92 (1H, b, OH), 6.51 (1 H, d, J= 7.7, N=CH), 3.05 (1 H, sept,2CH), 1.56-1.34 (2H, m,3CH2), 1.04 (3H, d, CH3- ), 0.92 (3H, t, CH3-w).1H NMR (DMSO-d6, ppm): 10.59 (1H, b, OH), 6.43 (1H, d, J= 7.3, N=CH), 2.89 (1 H, sept,2CH), 1.40 (2H, sex,3CH2), 0.94 (3H, d, CH3- ), 0.82 (3H, t, CH3-w).

[0133] Example 13. Preparation of (NaphtvD-1 -acetaldehyde, precursor of Naphtyl-1 - acetaldoxime (Naphtox)

[0134] A stirred solution of methyl (l-naphthyl)-acetate (437 pL, 2.50 mmol) in anhydrous toluene (20 mL) was cooled to -80 °C. Then, a DIBAL-H solution (1.2M, 2.5 mL, 3.0 mmol, 1.2 eq) was added dropwise keeping the reaction temperature below -75 °C. The resulting solution was stirred at -80 C for 30 min. and then methanol (5 mL) was carefully added. The solution was vigorously stirred and allowed to reach room temperature. HCI (1N, 50 mL) was then added and the solution was stirred for additional 10 min. The organic phase was separated and the aqueous layer was extracted with toluene (2x 20 mL). The combined organic phases were washed with Na2CO3(20 mL) and dried with magnesium sulfate. The solvent was evaporated under vacuum yielding (l-naphtyl)-acetaldehyde (412 mg, 97 %.) as a colorless oil.1H NMR (CDCh, ppm): 9.79 (dd, J= 2.44, 1H, CHO), 7.92- 7.87 (m, 2H, H5nap, H8nap), 7.84 (d, J= 8.16, 1H, H4Nap), 7.58-7.50 (m, 2H, H6nap, H7nap), 7.48 (dd, J= 8.10, 7.06, 1H, H3Nap) 7.41 (d, J= 6.92 , 1H, H2Nap), 4.11 (d, J= 2.44, 2H, H2).13C NMR (CDCh, ppm): 199.73 (C1), 134.17 (C8.aNap) 132.45 (C1 Nap), 129.07 (C5Nap o C8Nap), 128.64 (C2Nap), 128.57 (C4Nap), 128.53 (C4.aNap), 126.87, 126.24, 125.80 (C3Nap), 123.69 (C5Nap o C8Nap), 48.51 (C2)

[0135] Example 14. Preparation of Naphtyl-1 -acetaldoxime (Naphtox)

[0136] To a stirred solution of naphtyl-1 -acetaldehyde (412 mg, 2.42 mmol) in toluene / H2O (1:1, 40 mL) aqueous hydroxylamine was added (50% aq. 888 pL, 14.5 mmol, 6 eq). The resulting solution was vigorously stirred at room temperature for 4 hours and then the organic phase was separated. The aqueous layer was extracted with toluene (50 mL). The combined organic phases were dried with magnesium sulfate. The solvent was evaporated under vacuum yielding naphtyl-1 -acetaldoxime (352 mg, 78% rdto.) as a solid. Z-isomer,78 %:1H NMR (CDCh, ppm): 7.96 (1 H, d, J= 7.7, H8Nap), 7.91-7.85 (1 H, m, H5Nap), 7.79 (d, J=8.0, 7H, H4Nap), 7.58-7.48 (2H, m, 7.54 H7Nap, 7.51 H6Nap), 7.44 (1 H, t, J= 7.0, H3Nap), 7.39 (1 H, dd, J= 7.0, 1.2, H2Nap), 6.89 (1 H, t, J= 5.20, N=CH), 4.17 (2H, d, J= 5.2, CH2).13C NMR (CDCh, ppm): 151.2 (C1 , N=CH), 134.1 (C4aNap), 133.0 (C1Nap), 132.1 (C8aNap), 129.0 (C4Nap), 127.9 (C5Nap), 127.2 (C2Nap) 126.5 (C7Nap), 126.06 (C6Nap), 125.7 (C3Nap), 123.8 (C8Nap), 29.6 (C2)

[0137] E-isomer, 22 %:1H NMR (CDCh, ppm): 8.05 (d, J= 8.3, 1 H, H8Nap), 7.91-7.85 (1 H, m, H5Nap), 7.79 (7H, d, J=8.0, H4nap), 7.65 (1 H, t, J= 6.2, N=CH), 7.58-7.48 (2H, m, H7Nap, H6Nap), 7.44 (1 H, t, J= 7.0, H3Nap), 7.39-7.36 (1 H, m, H2Nap), 3.99 (2H, d, J= 6.1, CH2)13C NMR (CDCh, ppm): 150.8 (C1 , N=CH), 134.1 (C4aNap), 132.5 (C1 Nap), 132.1 (C8aNap), 129.0 (C4Nap), 128.0 (C5Nap), 127.0 (C2Nap), 126.5 (C7Nap), 126.0 (C6Nap), 125.7 (C3Nap), 123.8 (C8Nap), 33.6 (C2). [M+H+]: 186.0931 (calc, for CI2HI2NO+: 186.0919)

[0138] Example 15. Preparation of of 2,4-diclorophenoxyacetaldoxime (2,4 Ox)

[0139] A solution of 2,4-dichlorophenol (2.018 g, 12.57 mmol) and NaOH (503 mg, 12.57 mmol) in metanol (10 mL) was stirred for 5 min. and then the solvent was evaporated under reduced pressure. DMF (10 mL) and 2-bromoacetaldehyde diethylacetal (1.57 mL, 10.47 mmol) were added and the resulting solution was stirred at 100 °C for 5 h. The resulting mixture was allowed to cool and poured over a NaHCOs sat. (100 mL). The resulting solution was extracted with diethyl ether (3 x 50 mL).The organic phases were gathered and washed with an aqueous solution of NaOH (1 M, 50 mL) and H2O (50 mL) and finally treated with magnesium sulfate. The solvent was distilled by rotary evaporation to obtain a mixture of 2-bromoacetaldehyde diethylacetal and 2,4-dichlorophenoxyacetaldehyde diethylacetal that was purified by column chromatography (SiO2, Hexanes: Ethyl acetate 1 / 5) and dried under vacuum to obtain 2,4-dichlorophenoxyacetaldehyde diethylacetal (1100 mg, 32%).

[0140] 2,4-dichlorophenoxyacetaldehyde diethylacetal (278 mg, 1.00 mmol) was treated at 100 °C during 15 h with a solution of H2SO4 (1M, 3.0 mL). The solution was allowed to cool and NaHCCh sat. (25 mL) was added. The aqueous phase was extracted with diethyl ether (3 x 30 mL), the organic phases were gathered and treated with magnesium sulfate. The solvent was distilled to obtain a mixture of 2,4-dichlorophenoxyacetaldehyde and its hydrate. The mixture 2,4-dichlorophenoxyacetaldehyde diethylacetal was dissolved in a mixture of toluene / H2O (40mL, 1 :1) and hydroxylamine (50 % aq., 600 mL, 9.8 mmol) was added. The solution was stirred at room temperature for 6 hours and then the toluene was separated. The aqueous phase was extracted with toluene (2 x 20 mL) and the cumulative organic phases were dried with magnesium sulfate and the solvent was distilled under vacuum to obtain 2,4-chorophenoxyacetaldoxime (112 mg, 51%).

[0141] Z-isomer 94%:1H NMR (CDCh, ppm): 7.51 (1 H, b, OH), 7.40 (1 H, d, J= 2.6 Hz, H3ph), 7.19 (1 H, dd, J= 8.8, 2.5 Hz, H5ph), 7.05 (1 H, t, J= 3.6 Hz, N=CH), 6.84 (1 H, d, J= 8.9 Hz, H6ph) ,4.95 (2H, d, J= 3.7 Hz, CH2),13C NMR (CDCh, ppm): 152.6 (C1ph), 149.2 (C1 , N=CH), 130.5 (C3ph), 127.9 (C5ph), 126.9, 124.1 (C2ph, C4ph), 114.2(C6ph), 63.2 (C2, CH2).

[0142] E isomer 6%:1H NMR (CDCh, ppm): 7.63 (1 H, td, J= 5.5, 2.1 Hz, N=CH), 7.52 (1 H, b, NH), 7.38 (1 H, d, J= 2.6 Hz, H3ph), 7.19 (1 H, dd, J= 8.8, 2.5 Hz, , H5ph), 6.81 (1 H, d, J= 8.8 Hz, H6ph) ,4.71 (2H, d, J= 4.6 Hz, CH2).

[0143] Example 15: Administration of aldoximes to plants

[0144] M. truncatula Gaertn. ecotype Jemalong A17 plants were grown as described by Buezo et al.. Plants were grown in a growth chamber for 4 days at a day / night temperature of 24.5 / 22 °C, with 80% relative humidity, a 16 / 8 h day / night photoperiod and 70 pmol m’2s’1of photosynthetically active radiation. Harvesting was always conducted 6 hours after the light period onset to avoid circadian rhythm effects. Five randomly selected plants from different pots were collected; shoots and roots were separated, weighed, and then frozen in liquid N2, being stored at -80 °C for further analyses.

[0145] 100 - 200 pM lAOx was diluted in DMSO and was added to each of either solid (agar containing) or liquid media once the temperature dropped below 60 °C. As lAOx was diluted in DMSO, 0.25 pL / mL of DMSO was added to each non-hormone (control) medium to ensure that all treatments contained the same amount of DMSO. The rest of aldoximes were administered in the same way as lAOx.

[0146] Example 16: The aldoximes are able to release NO catalyzed by POD In vitro, which can be detected using DAF-FM as fluorescent specific probe for the detection of NO

[0147] It was evaluated the production of NO catalyzed by horseradish peroxidase (POD), and detected as fluorescence with the DAF-FM probe, using different aldoximes as substrates (1 H-lndol-3-acetaldoxime, lAOx; 1 H-lndol-3-carboxaldoxime, IMOx; 2-(2,3-dihidro-1 H- inden-3-il)-acetaldoxime, IDOx; benzaldoxime, BenzOx; 2-(4-hidroxiphenyl)-acetaldoxime, PhenOx), or different metabolites in the IAA synthetic pathway as IAA (indolil-3-acetic acid), Tryptofane (Trp), N-hidroxyl-tryptamine (NHT). The substrates Tryptophan (Trp) and IAA were used as a control as they were indolic molecules as lAOx but with no oxime group in it. Reactions were performed in black 96-well plate in a total volume of 300 pL. Each reaction contained 200 pM of each aldoxime or equivalent substrate, 1 pM of each haeme protein and 0.5 pM 4-Amino-5-methylamino-2',7'-difluorescein (DAF-FM) in 100 mM Tris- HCI 7.4 pH buffer.

[0148] The fluorescence measurement was taken 3h after the reaction in a spectrofluorimeter (BioTek Synergy HT) at 485nm (ex) and 520 (em). Values are the mean of 4 replicates ± S.E. Asterisks indicate significant differences (P value <0.01) between substrates using Dunnett's one-way ANOVA and post hoc tests between each aldoxime and control.

[0149] The results indicated that the aldoximes studied released NO in in vitro reactions when DAF-FM was used as the specific fluorescent probe for NO detection, whereas controls with Trp or IAA (FIG. 1) did not release NO. The aldoxime lAOx is the most efficient of those tested in releasing NO, while the control, IAA, showed no effect in releasing NO. The reaction mixtures with aldoximes that contained no POD enzyme showed no release of NO (not shown).

[0150] Also, the aldoxime lAOx at different concentration was tested in the spectrofluorimetric assay with the same indicated conditions. This NO production was dependent on the lAOx concentration and the POD (FIG. 2A and 2B), which confirms the enzymatic oxidation of lAOx by POD to release NO. At low lAOx concentration (<75 pM), the plot showed a concentration-dependent rise of NO production (FIG. 2A).

[0151] Example 17: Capability of the aldoximes of the invention for reducing iron at different pHs

[0152] The iron reduction was evaluated at different pHs for the following aldoximes lAOx, IMOx, BenzOx, ButOx, and IAA. The reduction of Fe3+was measured using the Ferrozin assay. The activity was expressed as the increase of the absorbance at 562 mm after 30’. The values are the media of 3 repetitions ± S.E. Asterisks indicates significant differences (P value <0,001) between molecules and control using ANOVA tests of two factors and Bonferroni.

[0153] Different aldoximes were tested at neutral (7.4) and slightly acidic (5.5) pH and showed different iron reduction capacities (FIG 3). Thus, in the presence of oxygen and hemoglobins such as POD, an iron reduction capacity of between 0.2 and 0.6 absorbance arbitrary units was observed for the most active aldoximes after 2 h of incubation in 1 mL mixtures, depending on the pH of the reaction. 18: Production of NO in conditions of normoxia, anoxia or anoxia plus

[0154] The production of NO from lAOx under anoxia / normoxia or anoxia + superoxide conditions was evaluated. The reactions were carried out in airtight glass vials containing 2 mM aldoxime, 10 M hemoprotein as catalyst, 10 pM flavin and 100 mM Tris-HCI pH 7.4 in 1 mL. The remaining conditions were those shown in FIG.4(see brief description of drawings section). Values are the mean of 5 replicates ± S.E. Group letters indicate significant differences (P value <0.01) in unidirectional ANOVA test and Tukey's post hoc test. NO production under conditions of normoxia, anoxia, or anoxia plus superoxide, showed that superoxide may be a more important inducer of NO release than oxygen in the case of aldoximes such as lAOx (FIG. 5).

[0155] The determination of NO, nitrite and nitrate from lAOx as a result of the catalysis of different types of hemoproteins (rice Hb1 Lbll, and horseradish peroxidase -POD) was evaluated by chemiluminescence as indicated in FIG. 4. The results show that both anoxia and the enzyme iron-superoxide dismutase from cowpea very markedly inhibited NO production, evidencing that oxygen is necessary for the reaction and the superoxide radical functions as an intermediate in the catalytic process (FIG. 4). The involvement of superoxide radical was an intermediate of the reaction of NO production is important in animals and plants, because it shows that lAOx reaction catalyzed by POD captures superoxide radical, which is considered a toxic molecule produced in all living organisms. The superoxide radical in the reaction contributes to produce NO, which is a less active free radical which in many cases shows a non-toxic function and can signal an antioxidant response. In this way, the aldoxime can contribute to the disposal of the radical superoxide and to the release of a less toxic or antioxidant NO molecule. In some specific human pathologies, as for example during hipoxia-reperfusion condition during heart surgery, the production of superoxide during the reperfusion of the heart can induce a strong tissue damage. The use of aldoximes in pre-treatment for this condition may represent a way to importantly reduce that damage.

[0156] Therefore, the enzyme xanthine oxidoreductase (XDH), whose enzymatic activity produces superoxide radical, was tested. Thus NO2' production catalyzed by the xanthine oxidoreductase (XDH) enzyme from lAOx in normoxia, in the presence or not of the substrate xanthine or the reduced nicotinamide dinucleotide reductor (NADH), was evaluated. Reactions were performed in airtight glass vials containing 2 mM aldoxime, 10 pM XDH enzyme, 1 mM NADH and 100 mM Tris-HCI pH 7.4 in 1 mL. All other conditions as indicated in FIG. 4. Values are the mean of 4 replicates ± S.E. Group letters indicate significant differences (P value <0.01) in unidirectional ANOVA test and Tukey's post hoc test. The results show that when including XDH in the reaction mixture with lAOx as a substrate at neutral pH a higher NC>2' production under normoxic conditions was observed (FIG. 6) producing higher rates of NC>2' release in vitro than when catalyzed by POD (FIGs 4, 3).

[0157] Example 19: Production of NO using a mixture of flavines and hemoglobines as catalysts

[0158] The production of NO catalyzed by horseradish peroxidase (POD) using different aldoximes as substrate, as indicated in the FIG.7 (1 H-lndol-3-acetaldoxime, lAOx; 1 H- lndol-3-carboxaldoxime, IMOx; 2-(2,3-dihydro-1 H-inden-3-yl)-acetaldoxime, IDOx; benzaldoxime, BenzOx; 2-(4-hydroxyphenyl)-acetaldoxime, PhenOx), or various metabolites of the IAA (indole-3-acetic acid) biosynthesis pathway from Trp (Tryptophan, Trp; N-hydroxyl-tryptamine, NHT; IAA) was evaluated by chemiluminescence,. Reactions were performed in airtight glass vials containing 2 mM aldoxime or NHT or L-Arg or IAA, 10 pM hemoprotein as catalyst, 10 pM flavin and 100 mM Tris-HCI pH 7.4 in 1 mL. The remaining conditions are as indicated in FIG. 4. Values are the mean of 5 replicates ± S.E. Asterisks indicate significant differences (p-value <0.001) in unidirectional ANOVA test and Dunnett's, post hoc test using lAOx-Met + POD + RbF as control.

[0159] In our study, the addition of flavins, as riboflavin (RbF), flavin mononucleotide (FMN) or flavin adenine dinucleotide (FAD), increased the release of NO from lAOx, and also the NO2- and NOT catalyzed by POD (FIG. 8). Among flavins, RbF provoked the highest boost in either NO and NO27NO3 production from lAOx, followed by FMN and FAD in the presence of POD (FIG. 7a). This result is in line with the differences in their volumes, and their reduction capacities in the O2 / POD oxidation. In parallel, the addition of flavins to Hbl or Lbll and lAOx did not increase the direct NO production but increased the NO27NOT levels (FIG. 8). This suggests that Hbl and Lbll do not directly interact with lAOx, and they are producing NO27NOT secondarily. Since the highest NO production was obtained with POD and RbF, all the previously studied molecules, together with L-Arg — substrate of inducible NOS (inos) — and IAA were then tested for NO release and NO2' and NOs accumulation in the presence of these two compounds (FIG. 7a). The relative rates of NO production were consistent with those observed in the absence of added flavin (FIG. 1 and 10A) and confirmed that lAOx is by far the most efficient aldoxime for the oxidative production of NO under our assay conditions (FIG 1 and FIG. 7A). As expected, the molecules lacking a hydroxyl group at the oxime, as lAOx-Met, L-Arg and IAA showed no NO-release capacity in presence of POD and riboflavin in the in vitro reactions (FIG. 7A). A continuous production of NO for 2h was directly detectable from the reaction of POD+IAOx+RbF (FIG. 7B). To gain insight on the role of O2; potassium superoxide was added to the reaction, significantly increasing the NO and NO27NOs' derived from lAOx (FIG. 5).

[0160] In the case of riboflavin, it was so efficient that direct NO production was observed (usually if a low amount of NO is produced it is immediately oxidized to nitrite, so only this form was observed). Nitrate produced in the same reaction and also originating from oxidation of primary produced NO was also detected.

[0161] The results show that especially good results are obtained with flavines, and in particular using a combination of flavins with POD (see FIGs 8, 5 and 7).

[0162] 20. Effect of aldoximes on the of the root of

[0163] To study the in vivo function of the NO release from aldoximes, we externally supplemented Medicago truncatula plants with lAOx and synthetic aldoximes: IMOx, IDOx, BenzOx, ButOx and PhenOx, and lAOX-Met were grown and treated with the indicated aldoximes as described in example 13. The NO-producing aldoximes tested induced an increase in the number of lateral roots (FIGs. 9 and 10) due to the capacity of the aldoximes to produce NO. The capacity of the NO donors other than aldoximes to induce lateral root growth has already been shown. Our experiments show now that aldoximes, which produce NO strongly induce the lateral roots of the plants. The phenotype obtained in the M. truncatula roots with IMOx showed a remarkable inhibition of the primary root elongation along with a rise in secondary roots (FIGs. 9 and 10).

[0164] Example 21. lAOx can serve as a substrate to the reaction catalyzed by Nitric Oxide Synthase. lAOx was tested in a NOS-catalysed reaction. For inducible Nitric Oxide Synthase (iNOS) NO measurements 100 pM Ca2+, 18 pM (6R)-5,6,7,8-Tetrahydrobiopterin (BF ), 1 mM Nicotinamide adenine dinucleotide phosphate (NADPH), 10 pg of calmodulin bovine, 1 mM of lAOx or L-Arg and 1 pM of iNOS. Our data show that lAOx can serve as a substrate for iNOS enzyme to produce NO, but less efficiently than L-Arg (FIG. 11). This will be also important in animal systems, and emphasizes the close relation between aldoximes to NO.

[0165] Example 22. Effect of IDOx to protect Medicago truncatula plants from stress induced by ammonium nutrition as an exclusive source of nitrogen. Medicago truncatula plants were grown and treated with aldoximes as described in example 13, but the plants were grown either using nitrate or ammonium as a sole source of N. Plants were photographed at day 0, 4, 8, 12, and 14 to establish a dataset containing architectural descriptions of the root system. The quantification of both the primary root length and the number of secondary roots were performed with Image J (Schindelin et al., 2012 Fiji: An open-source platform for biological-image analysis. Nat Methods 9:676-682) and the SmartRoot plugin (Lobet et al., 2015 Root system markup language: Toward a unified root architecture description language. Plant Physiol 167:617- 627) as described (Esteban Raquel et al., 2016 Both free indole-3-acetic acid and photosynthetic performance are important players in the response of Medicago truncatula to urea and ammonium nutrition under axenic conditions. Front Plant Sci 7:140).

[0166] Treatments with exclusive ammonium nutrition induced a stress that can be tracked by the reduction in plant biomass when we compare respect to the nitrate exclusive nutrition. The aldoxime IDOx significantly enhanced the shoot biomass and also the total plant biomass under ammonium nutrition (FIG. 12), hence demonstrating the capacity to induce a relief of the stress. Also, IDOx aldoxime induced the elongation in the primary root during initial stages (4 days) and also at day 14th. Furthermore, IDOx strongly increased the number of lateral (secondary) roots at day 4 and at day 14thas shown in FIG. 13 and in FIG. 14. This modification of the root phenotype does not involve a greater weight of the root as the biomass of the root is not significantly changed (FIG. 12), but it seems to involve a higher functionality of the root. This capacity of the IDOx to increase the main root length and lateral root number will be important for the plant to tolerates stresses as drought or high temperatures.

[0167] Example 23: Capability of inhibiting the growth of cancer cells

[0168] The lAOx was tested in combination with POD for their ability to inhibit cancer cell growth using in vitro culture models of MCF-7 cancer cells (which are a model of breast cancer cells). For this purpose, the GI50 was determined, which is defined as the concentration of compound that would inhibit 50% of cell growth 48 h after fixation on a Petri dish.

[0169] To determine this value, the cells were seeded and within four hours, after fixation on the support were counted and allowed to grow for 48 h. Viability counting was performed using an assay with MTT (yellow) that passed to its formazan (purple) in a reaction with succinate dehydrogenase (mitochondrial Complex II). Thus, the control without compound at time 48 h scored 100 and the intermediate values provide inhibition percentages. Table 1 : Toxicity effect of compounds on cell growth of MCF-7 breast cancer cells and control breast cells measured as GI50 (Growth inhibition rate 50).

[0170] 10

[0171] In this example, we can observe that the lAOx shows a good inhibition rate of the MCF-7 as evidenced by the low GI50 found. The GI50 for lAOx is low enough to be considered an active compound but also is remarkable the inhibition rate of the mix of lAOx + Sulforaphane, and also the ratio MCF-7 / Controls cells.

[0172] Also, since NO is a molecule that signals programmed cell death or defense against pathogens or antioxidant defenses, these aldoximes are useful in the treatment of diseases such as cancer.

[0173] Example 24: Effectiveness of the compounds on bacterial growth and biofilm formation in gram positive and gram-negative bacteria

[0174] Observation of the effect of the compounds on the biofilm-forming ability of the grampositive bacterium Staphylococcus aureus was carried out as follows. A colony of a strain of S. aureus was cultured in 5 ml of TSB supplemented with 0.25% glucose (TSB-Gluc) overnight at 37°C with 200 rpm shaking, the culture was adjusted to an optical density (OD) of 600 nm at 1 and a 1 / 40 dilution was made in 200 pl of TSB-Gluc in each well of 96-well polystyrene plates (Thermo). In order to analyze the effect of the compounds during the biofilm formation process, the compounds were resuspended in DMSO at an appropriate concentration to add 2 pl of the diluted compound to each well and that it would remain at the final concentration at which one wanted to study. In control wells, 2 pl of DMSO was added.

[0175] These plates were incubated under static conditions at 37°C overnight, the medium was removed from the wells, and the wells were washed with water 3 times. To visualize the biofilm, 200 pl of the crystal violet dye were added, incubated for 5 minutes at room temperature, and then the wells were washed with water three times. Biofilm formation was observed qualitatively and quantitatively. Quantification of the biofilm formed in the wells was performed as follows: 200 pl of an 80:20 volume / volume ethanol / acetone mixture were added, incubated for 5 minutes at room temperature. The 200 pl were taken and placed in another 96-well plate where the amount of dye was measured by measuring the absorbance at 595 nm.

[0176] Two different strains of S. aureus were used: strain 15981, which forms a biofilm of a polysaccharide nature dependent on the PIA / PNAG exopolysaccharide, and strain V329, which forms a biofilm of a protein nature.

[0177] The effect of the compounds on the biofilm formation capacity of the gram-negative bacterium Salmonella Enteritidis, strain 3934, was also analyzed. A colony of S. Enteritidis 3934 strain was incubated in 5ml of LB overnight at 37°C with 200rpm shaking, the culture was adjusted to an optical density (OD) of 600 nm at 1 and a 1 / 40 dilution was made in 5 ml of LB in borosilicate tubes. In order to analyze the effect of the compounds during the biofilm formation process, the compounds were resuspended in DMSO at an adequate concentration to add 50 pl of the diluted compound to each well and that it would remain at the final concentration at which one wanted to study. In control tubes, 50 pl of DMSO was added. The tubes were incubated for 96h at room temperature under static conditions, and biofilm formation was qualitatively observed at the medium-air interface.

[0178] The effect of the compounds on bacterial growth was studied by taking the culture from the wells, in the case of S. aureus, or from the tube, in the case of S. Enteritidis, after the biofilm formation process and making serial decimal dilutions of that culture in PBS. One drop (5 pl) of these dilutions was placed on a plate, on TSA in the case of S. aureus and on LB agar in the case of S. Enteritidis, and the bacteria were grown overnight at 37°C. It was thus quantified whether there were the same number of bacteria in the wells / tubes in the presence of the compounds or in the presence of the DMSO diluent alone.

[0179] The results of inhibition of the bacterial growth are shown in the following Table 2. The term NaphtOx refers to NaphtOx produced after initial synthesis as a mixture of Z and E isomers (i.e., Z78:E22), while similarly Z82:E18 indicates 82% of the isomer Z and 18% of isomer E produced after recrystalization. The term lAOx refers to the lAOx provided after initial synthesis as a mixture of Z and E isomers. An enriched lAOx sample in Z isomer was obtained after recrystallization (Z-IAOx), while Z75:E25 indicates 75% of the isomer Z and 25% of isomer E. All the compounds indicated were capable of inhibiting biofilm formation in any of the pathogenic bacteria, and one of them were capable of inhibiting bacterial growth (FIGS. 15-18). The compound 2,4 Ox was most active inhibiting biofilm formation and also inhibiting bacterial growth. Some of the lAOx isomers were also remarkable active when compared to the rest of the compounds.

[0180] Table 2. Inhibition of the bacterial growth by aldoximes:

[0181] ‘Unstable results.

[0182] Citation List

[0183] Non-Patent Literature

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Claims

Claims1. Aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof able of releasing nitric oxide (NO) for use in the treatment of a condition or a disease which is sensible to the action of the NO, wherein the treatment comprises administering the aldoxime to a subject and releasing NO from the aldoxime through an in vivo enzymatically catalyzed redox reaction either at neutral or at physiological pH.

2. Aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof for use according to claim 1 , wherein the redox reaction is enzymatically catalyzed by a peroxidase.

3. Aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof for use according to any of the claims 1-2, wherein the condition or disease is selected from the group consisting of: vasodilation, hypertension, a cardiovascular disease, an acute episode of cardiovascular disease, a cancerous tumor, an infectious disease, inflammatory condition, pathogen infection, and in the prevention of oxidative burst that occurs in situations of ischemia-reperfusion in open heart surgery.

4. Aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof for use according to claim 3, wherein the acute episode of cardiovascular disease is a heart attack.

5. Aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof for use according to claim 3, wherein the cardiovascular disease, the cancerous tumor or the inflammatory condition are mediated by the anti-inflammatory effect of the NO.

6. Aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof for use according to claim 5, which is selected from the group consisting of:

7. Use of an aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof able of releasing nitric oxide (NO) to improve the muscle contractility capacity, ventilatory efficiency and sports performance.

8. In vivo use of an aldoxime or an acceptable salt for plants thereof or an isomer thereof capable of releasing nitric oxide (NO) in in vivo systems, for biological applications which are sensible on the action of the NO in plants, wherein the biological application is selected from the group consisting of plant rooting agent, plant structure modifier, stimulators of the tolerance to abiotic or biotic stress, plant protector from stress induced by ammonium nutrition as an exclusive source of nitrogen, _plants protector from hydric stress, and as germination inducer.

9. In vivo use of an aldoxime or an acceptable salt for plants thereof or an isomer thereof according to claim 8, wherein the plant rooting agent comprises lateral root development stimulation, root surface increasing agent, or both.

10. In vivo use of an aldoxime or an acceptable salt for plants thereof or an isomer thereof according to claim 8, wherein the plant structure modifier action is either modulation or the aerial structure and bearing.11 . In vivo use of an aldoxime or an acceptable salt for plants thereof or an isomer thereof according to any of the claims 8-10, wherein the aldoxime is selected from the group consisting of:

12. In vivo use of an aldoxime or an acceptable salt for plants thereof or an isomer thereof according to claim 11 , together with a compound that catalyzes the in vivo release of nitric oxide (NO) from the aldoximes.13.- In vivo use of an aldoxime or an acceptable salt for plants thereof or an isomer thereof according to claim 12, wherein the compound that catalyze the in vivo release of nitric oxide (NO) from the aldoximes is selected from the group consisting of a peroxidase, a flavin, a hemoglobin; and a nitric oxide synthase.

14. Compound selected from 2-(2,3-dihidro-1 H-inden-3-il) acetaldoxime (IDOx) of formula:and 2-(4-hidroxiphenyl)-acetaldoxime, (PhenOx), of formula:PhenOx15. A composition which is selected from: a pharmaceutical composition comprising a therapeutically effective amount of an aldoxime or a pharmaceutically acceptable salt thereof or an isomer thereof capable of releasing nitric oxide (NO) in vivo, together with pharmaceutically acceptable excipients or carriers; and a composition for plants comprising an effective amount of an aldoxime or an or an acceptable salt for plants thereof or an isomer thereof capable of releasing nitric oxide (NO) in vivo, together with appropriate carriers, wherein the effective amount is for stimulating fruit and root development and for modifying plant structure.

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