THE CHILEAN PLANTS DISCARIA CHACAYE, TALGUENEA QUINQUENERVIA (RHAMNACEAE), PEUMUS BOLDUS (MONIMIACEAE), AND CRYPTOCARYA ALBA (LAURACEAE) WERE EVALUATED AGAINST CODLING MOTH: CYDIA POMONELLA L. (LEPIDOPTERA: TORTRICIDAE) AND FRUIT FLY DROSOPHILA MELANOGASTER (DIPTERA: DROSOPHILIDAE), WHICH IS ONE OF THE MOST WIDESPREAD AND DESTRUCTIVE PRIMARY PESTS OF PRUNUS (PLUMS, CHERRIES, PEACHES, NECTARINES, APRICOTS, ALMONDS), PEAR, WALNUTS, AND CHESTNUTS, AMONG OTHER. FOUR BENZYLISOQUINOLINE ALKALOIDS (COCLAURINE, LAUROLITSINE, BOLDINE, AND PUKATEINE) WERE ISOLATED FROM THE ABOVE MENTIONED PLANT SPECIES AND EVALUATED REGARDING THEIR INSECTICIDAL ACTIVITY AGAINST THE CODLING MOTH AND FRUIT FLY. THE RESULTS SHOWED THAT THESE ALKALOIDS POSSESS ACUTE AND CHRONIC INSECTICIDAL EFFECTS. THE MOST RELEVANT EFFECT WAS OBSERVED AT 10 ?G/ML AGAINST D. MELANOGASTER AND AT 50 ?G/ML AGAINST C. POMONELLA, BEING THE ALTERATION OF THE FEEDING, DEFORMATIONS, FAILURE IN THE DISPLACEMENT OF THE LARVAE IN THE FEEDING MEDIUM OF D. MELANOGASTER, AND MORTALITY VISIBLE EFFECTS. IN ADDITION, THE DOCKING RESULTS SHOW THAT THESE TYPE OF ALKALOIDS PRESENT A GOOD INTERACTION WITH OCTOPAMINE AND ECDYSONE RECEPTOR SHOWING A POSSIBLE ACTION MECHANISM.

THE PURPOSE OF THIS STUDY WAS TO DETERMINE THE INHIBITORY CAPACITY OF CEANOTHANES TRITERPENES ISOLATE FROM CHILEAN RHAMNACEAE ON ACETYLCHOLINESTERASE (ACHE) AND BUTYRYLCHOLINESTERASE (BCHE) ENZYMES. SEVEN CEANOTHANES TRITERPENES WERE ISOLATED FROM AERIAL PARTS OF PLANT MATERIAL BY CLASSICAL PHYTOCHEMICAL METHODS OR PREPARED BY THE HEMISYNTHETIC METHOD. STRUCTURES WERE DETERMINED BY THE SPECTROSCOPIC METHOD (1H-NMR AND 13C NMR) AND MASS SPECTROMETRY (MS). ACHE AND BCHE ACTIVITY WERE DETERMINED BY THE ELLMANN METHOD FOR ALL COMPOUNDS. ALL TESTED COMPOUNDS EXERTED A GREATER AFFINITY TO ACHE THAN TO BCHE, WHERE COMPOUND 3 HAS AN IC50 OF 0.126 UM FOR ACHE AND OF >500 UM TO BCHE. KINETIC STUDIES INDICATED THAT ITS INHIBITION WAS COMPETITIVE AND REVERSIBLE. ACCORDING TO THE MOLECULAR COUPLING AND DISPLACEMENT STUDIES OF THE PROPIDIUM IODIDE TEST, THE INHIBITORY EFFECT OF COMPOUND 3 WOULD BE PRODUCED BY INTERACTION WITH THE PERIPHERAL ANIONIC SITE (PAS) OF ACHE. THE COMPOUNDS TESTED (1?7) SHOWED AN IMPORTANT INHIBITORY ACTIVITY OF ACHE, BINDING TO PAS. THEREFORE, INHIBITORS THAT BIND TO PAS WOULD PREVENT THE FORMATION OF THE ACHE-A? COMPLEX, CONSTITUTING A NEW ALTERNATIVE IN THE TREATMENT OF ALZHEIMER?S DISEASE (AD).

CARBOHYDRATES FROM LIGNOCELLULOSIC FEED CAN BE RELEASED BY BASIDIOMYCETE FUNGI FOR RUMINAL FERMENTATION. THIS STUDY AIMED TO EVALUATE THE BIOCONVERSION OF HAY OF RYEGRASS-FESCUE (LOLIUM PERENNE?FESTUCA ARUNDINÁCEA) BY SOLID STATE FERMENTATION WITH PLEUROTUS OSTREATUS, TO OBTAIN SUPERIOR QUALITY HAY. AFTER ONLY 14 DAYS OF FERMENTATION, CRUDE PROTEIN (CP) (4.73 TO 5.16%), AND NON-FIBROUS CARBOHYDRATES (NFC) (20.84 TO 25.04%) INCREASED, WHILE NEUTRAL DETERGENT FIBER (NDF) (68.72 TO 64.87%) AND ACID DETERGENT LIGNIN (5.88 TO 1.98%) DECREASED. THE ENZYMATIC BIODEGRADATION CARRIED OUT BY P. OSTREATUS WAS VERIFIED, THROUGH MEASUREMENTS OF ENZYMATIC ACTIVITY. LIGNIN PEROXIDASE (LIP) AND LACCASE (LAC) REACHED THE HIGHER ACTIVITY ON DAY 14 (19.51 U/L AND 34.17 U/L, RESPECTIVELY), WHEREAS MANGANESE PEROXIDASE (MNP) DISPLAYED STABILITY UP TO 21 DAYS OF FERMENTATION (BETWEEN 6.54 AND 7.75 U/L). IN CONCLUSION, RESULTS INDICATE THAT LIGNOCELLULOSIC FEED BIOCONVERSION BY P. OSTREATUS IS PROMISING TO IMPROVE THE RUMINAL FERMENTATION OF FIBROUS FEEDSTOCKS AND 14 DAYS WERE CONSIDERED TO BE OPTIMAL FOR HAY FERMENTATION.

MANY SECONDARY METABOLITES FROM PLANTS ARE CONSTITUTIVE AND/OR INDUCED CHEMICAL DEFENSES THAT PLAY A ROLE IN PROTECTION AGAINST PHYTOPATHOGENIC ATTACK. TALGUENEA QUINQUENERVIA (RHAMNACEAE) IS A NOTORIOUS WEED THAT POSSESSES SEVERAL MEDICINAL APPLICATIONS SUCH AS AMERICAN TAXA OF RHAMNACEAE SPECIES ARE TOXIC TO INSECTS, FUNGI AND SEVERAL BACTERIA STRAINS. THESE EFFECTS HAVE BEEN ASSOCIATED WITH THE PRESENCE OF ALKALOIDS, PHENOLICS AND TERPENES. A METHANOLIC EXTRACT FROM THE ROOTS AFFORDED 9 TRITERPENES BELONGING TO LUPANE, OLEANANE, URSANE, STEROL AND CEANOTHANE GROUPS. OUR RESULTS INDICATE THAT THESE COMPOUNDS ARE INVOLVED IN INTERFERENCE OF SCLEROTIZATION AND MOULTING. THEY SHOWED SELECTIVE EFFECTS ON PRE-EMERGENCE METABOLISM OF SELECTED INSECTS INCLUDING LEPIDOPTERA AND DIPTERA, BUT NOT TO COLEOPTERA. THE RESULTS WERE FULLY COMPARABLE TO KNOWN NATURAL INSECT GROWTH INHIBITORS SUCH AS AZADIRACHTIN, GEDUNIN AND CEDRELA EXTRACT AND HAVE A POSSIBLE ROLE AS NATURAL INSECTICIDAL AGENTS. TO THE BEST OUR KNOWLEDGE, THIS IS THE FIRST REPORT OF THE ISOLATION OF CEANOTHANE-TYPE TRITERPENES FROM THIS PLANT SPECIES. THE LEVEL OF ACTIVITY ACHIEVED BY THE ASSAYED COMPOUNDS INDICATED INHIBITION IN A CONCENTRATION DEPENDENT MANNER WITH LD50 VALUES BETWEEN 41 AND 17 ?G/ML.

ALZHEIMER?S DISEASE (AD) IS A MULTIFACTORIAL AND FATAL NEURODEGENERATIVE DISORDER. ACETYLCHOLINESTERASE (ACHE) PLAYS A KEY ROLE IN THE REGULATION OF THE CHOLINERGIC SYSTEM AND PARTICULARLY IN THE FORMATION OF AMYLOID PLAQUES; THEREFORE, THE INHIBITION OF ACHE HAS BECOME ONE OF THE MOST PROMISING STRATEGIES FOR THE TREATMENT OF AD, PARTICULARLY CONCERNING ACHE INHIBITORS THAT INTERACT WITH THE PERIPHERAL ANIONIC SITE (PAS). CEANOTHIC ACID ISOLATED FROM THE CHILEAN RHAMNACEAE PLANTS IS AN INHIBITOR OF ACHE THROUGH ITS INTERACTION WITH PAS. IN THIS STUDY, SIX CEANOTHIC ACID DERIVATIVES WERE PREPARED, AND ALL SHOWED INHIBITORY ACTIVITY AGAINST ACHE. THE STRUCTURAL MODIFICATIONS WERE PERFORMED STARTING FROM CEANOTHIC ACID BY APPLICATION OF SIMPLE SYNTHETIC ROUTES: ESTERIFICATION, REDUCTION, AND OXIDATION. ACHE ACTIVITY WAS DETERMINED BY THE ELLMANN METHOD FOR ALL COMPOUNDS. KINETIC STUDIES INDICATED THAT ITS INHIBITION WAS COMPETITIVE AND REVERSIBLE. ACCORDING TO THE MOLECULAR COUPLING AND DISPLACEMENT STUDIES OF THE PROPIDIUM IODIDE TEST, THE INHIBITORY EFFECT OF COMPOUNDS WOULD BE PRODUCED BY INTERACTION WITH THE PAS OF ACHE. IN SILICO PREDICTIONS OF PHYSICOCHEMICAL PROPERTIES, PHARMACOKINETICS, DRUG-LIKENESS, AND MEDICINAL CHEMISTRY FRIENDLINESS OF THE CEANOTHANE DERIVATIVES WERE PERFORMED USING THE SWISS ADME TOOL.

CYCLOPEPTIDE ALKALOIDS WITH DIFFERENT BIOLOGICAL ACTIVITIES ARE PRESENT IN PLANTS OF THE FAMILY RHAMNACEAE. PLANTS OF THIS FAMILY GROW IN A SYMBIOTIC RELATIONSHIP WITH AEROBIC GRAM-POSITIVE ACTINOMYCETES BELONGING TO THE GENUS FRANKIA. THIS GOAL OF THIS RESEARCH WAS A STUDY OF THE COMPARATIVE PROFILE OF ALKALOIDS PRESENT IN DISCARIA CHACAYE AND TO ESTABLISH A CONNECTION BETWEEN THE PRESENCE OR ABSENCE OF FRANKIA SP. AND THE ALKALOIDS. IN ADDITION, INSECTICIDAL ACTIVITIES OF THE ALKALOIDAL EXTRACT WERE EXAMINED. A TOTAL OF 24 ALKALOIDS WERE IDENTIFIED, OF WHICH 12 HAVE A BENZYLISOQUINOLINE SKELETON, 9 WERE CYCLOPEPTIDES, 2 ISOQUINOLINES, AND 1 APORPHINE. THE PRESENCE OF CYCLOPEPTIDE ALKALOIDS IS ASSOCIATED WITH FRANKIA NODULES IN THE PLANT ROOT. THE ALKALOID EXTRACTS SHOWED INSECTICIDAL ACTIVITY WITH MORTALITY DOSE-DEPENDENCE AND LD50 VALUES BETWEEN 44 TO 71 ?G/ML.

IN ORDER TO FIND MOLECULES OF NATURAL ORIGIN WITH POTENTIAL BIOLOGICAL ACTIVITIES, WE ISOLATE AND SYNTHESISE COMPOUNDS WITH AGAROFURAN SKELETONS (EPOXYEUDESMANES). FROM THE SEEDS OF MAYTENUS DISTICHA AND MAYTENUS MAGELLANICA WE OBTAINED SIX DIHYDRO-?-AGAROFURANS, AND BY MEANS OF THE ROBINSON ANNULATION REACTION WE SYNTHESISED FIVE COMPOUNDS WITH THE SAME SKELETON. THE STRUCTURES WERE ESTABLISHED ON THE BASIS OF NMR, IR, AND MS. THE EVALUATED COMPOUNDS SHOWED INHIBITORY ACTIVITY ON THE ACETYLCHOLINESTERASE ENZYME AND ON THE COX ENZYMES. COMPOUND 4 EMERGED AS THE MOST POTENT IN THE ACETYLCHOLINESTERASE INHIBITION ASSAY WITH IC50 17.0 ± 0.016 ?M ON ACETYLCHOLINESTERASE (ACHE). THE COMPOUNDS EVALUATED WERE SHOWN TO BE SELECTIVE FOR ACHE. THE MOLECULAR DOCKING, AND THE PROPIDIUM DISPLACEMENT ASSAY SUGGESTED THAT THE COMPOUNDS DO NOT BIND TO THE ACTIVE SITE OF THE ENZYME ACHE, BUT RATHER BIND TO THE PERIPHERAL ANIONIC SITE (PAS) OF THE ENZYME, ON THE OTHER HAND, THE NATURAL COMPOUND 8, SHOWED THE BEST INHIBITORY ACTIVITY ON THE COX-2 ENZYME WITH AN IC50 VALUE OF 0.04 ± 0.007 ?M. THE PHARMACOKINETIC PROFILE CALCULATED IN SILICO USING THE SWISSADME PLATFORM SHOWS THAT THESE MOLECULES COULD BE CONSIDERED AS POTENTIAL DRUGS FOR THE TREATMENT OF NEURODEGENERATIVE DISEASES SUCH AS AD.

THE LOSS OF AGRICULTURAL PRODUCTS BY PESTS AND MICROORGANISMS IS INTENSELY FOUGHT WITH SYNTHETIC PRODUCTS THAT CAN POSE A SERIOUS THREAT TO THE ENVIRONMENT, HEALTH, AND FOODS. THE USE OF PLANT-DERIVED PRODUCTS IN THE MANAGEMENT OF THESE ORGANISMS IS A SUSTAINABLE AND SAFE ALTERNATIVE. IN THIS STUDY, WE INVESTIGATED FABIANA IMBRICATA, A VERY WELL-KNOWN SPECIES OF SOLANACEAE FOUND IN THE PATAGONIAN REGION. THE TOXICOLOGICAL EFFECTS OF ITS FRACTIONS WERE TESTED AGAINST DROSOPHILA MELANOGASTER AND TENEBRIO MOLITOR. THESE FRACTIONS WERE ALSO SCREENED FOR REPELLENCY ON ACANTHOSCELIDES OBTECTUS AND ANTIMICROBIAL ACTIVITY. ADDITIONALLY, THE MORTALITY ASSAY OF D. MELANOGASTER LARVAE REVEALED HIGHER ACTIVITY FOR THE STEM (LC50 VALUE OF 7.5 ?G/ML) AND LEAF (LC50 VALUE OF 19.8 ?G/ML) PETROLEUM BENZINE FRACTIONS. TENEBRIO MOLITOR LARVAE NEEDED TO BE EXPOSED TO A HIGH CONCENTRATION OF STEM FRACTIONS (200 AND 300 ?G/ML) TO OBSERVE ABOUT 50 AND 90% MORTALITY, RESPECTIVELY. MOREOVER, REPELLENCY AFTER 4 H OF EXPOSURE WAS MAINLY OBSERVED WITH THE STEM DICHLOROMETHANE FRACTION (CONC. 100 ?G/ML). THE ACETATE FRACTION OF THE STEM (FE-C) WAS RESPONSIBLE FOR THE GREATEST GROWTH INHIBITION OF THE TESTED BACTERIA (ESCHERICHIA COLI, LISTERIA MONOCYTOGENES, L. INNOCUA, SALMONELLA SP., AND S. AUREUS). BASED ON THE FINDINGS, STEM FRACTIONS WERE SELECTED FOR STUDIES OF TOXICITY IN NON-TARGET ORGANISMS AND SYSTEMIC EFFECTS ON PROTEINS AND DETOXIFICATION ENZYMES FROM T. MOLITOR LARVAE. THE FRACTIONS TESTED AGAINST ADULT BEES WERE SAFE UP TO 500 ?G/ML FOR 4 H OF EXPOSURE AND 48 H OF OBSERVATION. TOTAL PROTEINS AND ENZYMES ACHE, GST/GSH, AND CARBOXYLASE WERE SIGNIFICANTLY REDUCED, MAINLY WITH FE-C. IN CONTRAST, ACP AND ALP WERE UP-REGULATED, WITH THE SAME FRACTION AS THE MAJOR INDUCER. THE CHEMICAL PROFILES OF THE FRACTIONS ANALYZED BY GC-MS AND NMR SHOWED THAT FE-C HAD SCOPOLETIN AS THE MAIN COMPONENT. THEREFORE, WE SELECTED THIS COMPOUND FOR IN SILICO ANALYSIS OF ACHE DOCKING, QUALITATIVE AND QUANT

INTRODUCTION: TALGUENEA QUINQUINERVIA HAS BEEN USED IN FOLK MEDICINE AND TO DYE WOOL, AND THE MAIN CONSTITUENTS ARE ALKALOIDS AND TRITERPENES. IDENTIFICATION OF THESE TYPE OF COMPOUNDS IN THIS SPECIE IS A NECESSARY STEP TO UNDERSTAND THE BIOLOGICAL PROPERTIES. OBJECTIVE: TO EVALUATE THE RELATIONSHIP BETWEEN THE CHEMICAL COMPOSITION OF ROOT FROM T. QUINQUINERVIA AND ITS INSECTICIDAL PROPERTIES USING LIQUID CHROMATOGRAPHY ELECTROSPRAY IONISATION TANDEM MASS SPECTROMETRY (LC-ESI-MS/MS). METHODS: ALKALOIDS WERE EXTRACTED USING SOXHLET EXTRACTION WITH METHANOL. TOTAL EXTRACT WAS PARTITIONED AT PH 2 AND 12 TO ENRICH ALKALOID CONSTITUENTS AND TO REMOVE INTERFERENCES. THE SEPARATION OF ALKALOIDS IN THE TALGUENEA EXTRACT WAS PERFORMED ON A C18 COLUMN USING GRADIENT ELUTION AND THEIR TANDEM MASS SPECTRA WERE OBTAINED BY QUADRUPOLE TIME-OF-FLIGHT TANDEM MASS SPECTROMETRY (QTOF-MS/MS) TO PERFORM ACCURATE MASS MEASUREMENTS OF FRAGMENT IONS FOR THE ALKALOID CONSTITUENTS. RESULTS: SEVERAL TYPES OF ALKALOIDS WERE SEPARATED AND IDENTIFIED BY LC-ESI-MS/MS. THE STRUCTURAL ASSIGNMENT OF INDIVIDUAL ALKALOIDS WAS PERFORMED BASED ON CONVERGENCE OF MS/MS SPECTRAL DATA, PH PARTITIONING BEHAVIOUR, LC RETENTION BEHAVIOUR, AND ACCURATE MASS MEASUREMENTS. THE PH PARTITION OF THE EXTRACT PROVIDED STRUCTURAL INFORMATION ABOUT UNKNOWN ALKALOIDS EXTRACTED FROM T. QUINQUINERVIA. A TOTAL OF 20 COMPOUNDS WERE IDENTIFIED AND TENTATIVELY CHARACTERISED, AND OF THESE 15 ALKALOIDS WERE REPORTED FOR THE FIRST TIME IN THE INVESTIGATED T. QUINQUINERVIA. CONCLUSION: THE CHEMICAL PROFILING OF ALKALOIDS IN T. QUINQUINERVIA WITH DIFFERENT ORIGINS WAS PERFORMED FOR THE FIRST TIME AND PROVIDED DIAGNOSTIC IONS FOR DIVERSE ALKALOIDS IN T. QUINQUINERVIA. INSECTICIDAL ACTIVITY OBSERVED CAN BE EXPLAIN BY THE PRESENCE OF ALKALOID AND PENTACYCLIC TRITERPENES ON THE FRACTIONS ASSAYED.

THIS WORK EVALUATED THE INSECTICIDAL, ANTIFEEDANT AND ACHE INHIBITORY ACTIVITY OF COMPOUNDS WITH EUDESMANE SKELETON. THE INSECTICIDAL ACTIVITY WAS TESTED AGAINST LARVAE OF DROSOPHILA MELANOGASTER AND CYDIA POMONELLA, THE COMPOUNDS 3 AND 4 WERE THE MOST ACTIVE (LC50 OF 104.2 AND 106.7 ?M; 82.0 AND 84.4 ?M, RESPECTIVELY). LIKEWISE, THE MENTIONED COMPOUNDS WERE THOSE THAT SHOWED THE HIGHEST ACETYLCHOLINESTERASE INHIBITORY ACTIVITY, WITH IC50 OF 0.26 ± 0.016 AND 0.77 ± 0.016 ?M, RESPECTIVELY.