Alzheimer’s is the most common cause of dementia worldwide and seriously affects patients’ daily tasks. Plant endophytic fungi are known for providing novel and unique secondary metabolites with diverse activities. This review focuses primarily on the published research regarding anti-Alzheimer’s natural products derived from endophytic fungi between 2002 and 2022. Following a thorough review of the literature, 468 compounds with anti-Alzheimer’s-related activities are reviewed and classified based on their structural skeletons, primarily including alkaloids, peptides, polyketides, terpenoids, and sterides. The classification, occurrences, and bioactivities of these natural products from endophytic fungi are summarized in detail. Our results provide a reference on endophytic fungi natural products that may assist in the development of new anti-Alzheimer’s compounds. Alzheimer’s disease (AD) is a progressive neurodegenerative disorder and the most common cause of dementia worldwide that affects memory, thinking, and behavior and even interferes with daily tasks. The abnormal accumulation of beta-amyloid and phosphorylated tau proteins and nerve cell degeneration are deemed to play key roles in Alzheimer’s disease ,2. AccorEndophytic fungi were first identified in plants in 1809 ,11,12. β-site amyloid precursor protein-cleaving enzyme 1 (BACE1) inhibition, and their antioxidant activities. So far, the secondary metabolites of plant endophytic fungi with anti-Alzheimer’s activities have not been summarized. This review mainly focuses on the classification, occurrences, and bioactivites of the secondary metabolites derived from endophytic fungi.In this review, a comprehensive survey of approximately 468 compounds with anti-Alzheimer’s-related activities derived from endophytic fungi from 2002 to 2022 is performed. These compounds are classified by their structure skeleton and mainly include alkaloids, peptides, polyketides, terpenoids, and sterides. The most investigated activities of these metabolites are the inhibition of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), neuroprotection, Xylaria sp. collected from the leaves of Casearia sylvestris showed cytochalasins B–D (1–3) , 5,6-dehydro-7-hydroxy cytochalasin E (5), Δ6,12-isomer of 5 (6), and rosellichalasin (7). Compounds 4–7 showed weak anti-AChE activities with IC50 from 110.9 to 176.0 μM  (58), cyclo- (59), cyclo-(L-Leu-L-Pro) (60), cyclo-(R-Leu-R-Pro) (61), cyclo-(L-Ile-L-Pro) (62), and cyclo-(4-methyl-R-Pro-S-Nva) (63). None showed significant neuroprotection against H2O2-mediated cytotoxicity for HT22 cells at 100 μM pyridine 1-oxide group. The cell activities of 123 were 76% at 40.0 μM, which showed neuroprotection against corticosterone-mediated PC12 cell injury with a concentration-dependent effect within the scope of 5.0–40.0 μM -5,7-dimethoxynaphtho thiophene-4,9-dione (227), as well as anhydrojavanicin (228), 8-O-methyljavanicin (229), botryosphaerone D (230), and 6-ethyl-5-hydroxy-3,7-dimethoxynaphthoquinone (231). The IC50 of 228 for anti-AChE activity was 2.01 μM  (264), were isolated from Microsphaeropsis olivacea related to Pilgerodendron uviferum (D. Don) Florin (“Cipres de las Guaitecas”). The IC50s of 260–262 for AChE inhibition were 83, 37, and 89 µg/mL, respectively [A new enalin analog, 7-hydroxy-2,4dimethyl-3(2H)-benzofuranone (ectively .Corynespora cassiicola L36 from Lindenbergi philippensis (Cham.) resulted in the observation of corynesidones A (265) and B (266), corynether A (267), and a diaryl ether (268). Corynesidone B (266) showed scavenging DPPH activity with IC50 = 22.4 µM [An intensive chemical assay for  22.4 µM .Penicillium citrinum from Bruguiera gymnorrhiza led to the identification of (Z)-7,40-dimethoxy-6-hydroxy-aurone-4-O-b-glucopyranoside (269) and -1-(40-hydroxyl-phenyl)-3,4-dihydro-3,4,5-trimethyl-1H-2-benzopyran-6,8-diol (270). Compound 269 showed stronger neuroprotection than did 270 with respect to MPP+-mediated PC12 cell damage. The mechanism of 269 involved improving cell viability and mitochondrial membrane potential, inhibiting caspase-3 and caspase-9 expression and reducing DNA fragment formation [An investigation of ormation .271) was identified in the endophytic fungus Pestalotiopsis microspora isolated from Terminalia morobensis. Compound 271 exhibited potent scavenging OH activity at 0.22 mM [The isobenzofuranone isopestacin ( 0.22 mM .272) identified in the endophyte Cochliobolus kusanoi from Nerium oleander L demonstrated a 50% scavenging DPPH capacity at 0.194 mM [Oosporein (0.194 mM .Sporothrix sp. (#4335) revealed the isolation of sporothrins A–C (273–275) and sporothrin C (276), 1-hydroxy 8-methoxy-naphthalene (277), and 1,8-dimethoxy-naphthalene (278)  . Compoun 1.05 μM ,68.279–281), were collected from Alternaria sp. interrelated with the stem of Maianthemum bifolium. Compound 281 alone exhibited anti-AChE capacity with IC50 at 68.3 μM [Three novel aromatic polyketide dimers, bialternacins B–D ( 68.3 μM .Phomopsis sp. NXZ-05 related to the twigs of Camptotheca acuminata DECNE. (Nyssaceae). revealed seven compounds: 8-O-acetylmultiplolide A (282), 8-O-acetyl-5,6-dihydro-5,6-epoxymultiplolide A (283), 5,6-dihydro-5,6-epoxymultiplolide A (284), 3,4-deoxy-3,4-didehydromul-tiplolide A (285), (4E)-6,7,9-trihydroxydec-4-enoic acid (286), methyl (4E)-6,7,9-trihydroxydec-4-enoate (287), and multiplolide A (288)  . The eva10 mg/mL .Cladosporium cladosporioides MA-299, an endophytic fungus from the mangrove plant leaves of Bruguiera gymnorrhiza, contributed to the isolation of new compounds 5R-hydroxyrecifeiolide (289), 5S-hydroxyrecifeiolide (290), ent-cladospolide F (291), cladospolide G (292), and cladospolide H (293) together with known compounds iso-cladospolide B (294) and pandangolide 1 (295). Among them, 291 alone exhibited strong AChE inhibition with an IC50 value of 40.26 µM [Detailed chemical research on 40.26 µM .Aspergillus flavus cf-5 from the red alga Corallina officinalis revealed the isolation of the new compound -10,11-dihydroxyoctadeca-8,12-dienoic acid (296), which had a weak AChE inhibitory capacity with a rate of 10.3% at 100 µg/mL [A study on 00 µg/mL .297) and hexylitaconic acid (298) were identified from a new endophyte Curvularia sp., which was discovered on the stem bark of Rauwolfia macrophylla. The IC50s of 297 and 298 for inhibiting AChE were 1.93 and 1.54 μM, respectively [2′-Deoxyribolactone  and B (300). The IC50 of 299 for AChE inhibition was 12.63 μM [A chemical assay for 12.63 μM .301) was isolated from the endophyte Curvularia sp. G6-32 from the plant Sapindus saponaria L. It showed anti-DPPH and anti-ABTS activities with inhibitory rates of 22.5% and 62.7%, respectively [The compound E-G6-32 ; one new benzofuran derivative, daldiniol F (309); one new naphthol derivative, daldiniol G (310); and two known analogs, 4-hydroxy-3-(3-methylbut-3-en-l-ynyl)benzyl alcohol (306) and methoxy-3-(3-methylbut-3-enl-ynyl)benzyl alcohol (307). The IC50s of 306, 307, 309, and 310 for anti-BChE activities were 6.93 ± 0.71, 16.00 ± 0.30, 23.33 ± 0.55, and 15.53 ± 0.39 μM, respectively [The extensive investigation of ectively .311–313), along with 4-hydroxy-3-(3′-methylbut-3′-en-1′-ynyl)-benzoic acid (314) and 4-hydroxy-3-prenyl-benzoic acid (315), were reported in the Saccharicola sp. of Eugenia jambolana. Compound 311 alone exhibited inhibition toward huAChE-ICER and eeAChE-ICER with IC50s of 0.076 ± 0.01 and 0.0047 ± 0.0009 mg/mL, respectively [Three new oxygenated cyclohexanoids, speciosins U–W , as well as the known compounds stemphyperylenol (317), 3(ζ)hydroxy-octadeca-4(E),6(Z)-dienoic acid (318), E-7,9-diene-11-methenyl palmitic acid (319), p-hydroxybenzonic acid (320), and benzoic acid (321)  . Compounnd 80 μM .322) and B (323), were separated from the endophyte Trematosphaeria terricola isolated from desert plant Artemisia desertorum. Compound 322 alone dispalyed neuroprotection for corticosterone-induced PC12 cell damage at 6.25 μM [Two unusual dimers, trematosphones A , together with three known compounds: xenofuranone B (325), linoleic acid (326), and 2-hexenoic acid (327). Compound 326 showed potential neuroprotective activities toward glutamate-injured PC12 cells with an EC50 of 33.9 μM. Compound 324 showed no neuroprotective activity at 40 μM, and 325 and 327 even exhibited weak cytotoxicity at 40 μM [A study on at 40 μM .328) was found in Cochliobolus lunatus SCSIO41401. This compound displayed mild AChE inhibition with IC50 at 2.5 ± 0.21 μM, while the IC50 for the active control of huperzine A was 0.30 ± 0.06 μM [A phthalide glycerol ether (329–334)   were sep 40.0 μM .Penicillium sp. sk14JW2P collected from the roots of Kandelia candel (L.) DRUCE revealed the existence of 13-hydroxypalitantin (335) and (+)-palitantin (336), which exhibited anti-AChE activities with IC50 values of 12 ± 0.3 and 79 ± 2 nM, respectively, while the IC50 for the positive control of huperzine A was 0.06 μM [Extensive research on Aspergillus sp. xy02 from a Thai mangrove Xylocarpus moluccensis uncovered seven new compounds, including -7,10-epoxysydonic acid (337), -7,10-epoxysydonic acid (338), -7,12-epoxysydonic acid (339), -7,12-epoxysydonic acid (340), 7-deoxy-7,14-didehydro-12-hydroxysydonic acid (341), (Z)-7-deoxy-7,8-didehydro-12-hydro-xysydonic acid (342), and (E)-7-deoxy-7,8-didehydro-12-hydroxysydonic acid (343), as well as five known compounds: (+)-1-hydroxyboivinianic acid (344), engyodontiumone I (345), (+)-sydonic acid (346), (+)-hydroxysydonic acid (347), and (−)-(7S)-10-hy-droxysydonic acid (348). Compound 348 alone displayed a moderate scavenging DPPH capacity with an IC50 of 72.1 μM [The intensive study of endophyte  72.1 μM .Phaeosphaeria sp. LF5 from the leaves of Huperzia serrata generated the identification of 3-(hydroxymethyl)-5-methylfuran-2(5H)-one (349), aspilactonols G–I (350–352), and E-∆2-anhydromevalonic acid (353). Compound 352 alone exhibited anti-AChE activity with IC50 at 6.26 µM. The other compounds showed no activity at 100 µM [Intensive chemical research on 5H)-one 3, aspilacEpicoccum sp. YUD17002 and Armillaria sp. contributed to the discovery of armilliphatics A–C (354–356). The IC50 value of compound 354 for anti-AChE activity was 23.85 μM. The other compounds were inactive against AChE at 50 μM [Investigation into a co-culture of endophyte at 50 μM .357), orsellide F (358), orsellide A (359), globosumone B (360), and globosumone C (361) were obtained from Chaetoium sp. NF00754. The IC50 values for compounds 359 and 361 for anti-AChE activity were 7.34 and 7.67 µM, respectively [A rare 1-oxaspiro chaetospirolactone (ectively .362) and B (363)  [Two new compounds, asperterpenols A ( B (363) , with a >100 μM) .R,5R,6R,7R,10S)-1,6-dihroxyeudesm-4(15)-ene (364) was identified from Alternaria alternate interrelated with the leaves of Psidium littorale Raddi. This compound was inactive for neuroprotective activity toward glutamate-injured PC12 cells at 40 and 80 μM [The new compound  and B (366), and two new drimane-type sesquiterpenes, ustusol D (367) and ustusol E (368), along with known compounds 12-hydroxyalbrassitriol (369), 2-hydroxyalbrassitriol (370), deoxyuvidin B (371), 3β,9α,11-trihydroxy-6-oxodrim-7-ene (372), 2α,11-dihydroxy-6-oxodrim-7-ene (373), and ustusol B (374). The AChE inhibition ratios of 365–374 were in the range of 17.56 ± 3.33 to 57.38 ± 4.51%. The IC50s of 369 and 370 for anti-AChE activities were 43.02 ± 6.01 and 35.97 ± 2.12 μM, respectively. The binding sites of 369 to the AChE catalytic pocket were Trp84, Gly117, Ser122, and Tyr121 residues, while 370 lay on Asp72 and Ser122 residues [The extensive chemical investigation of endophyte Pseudofusicoccum sp. J003 from the mangrove species Sonneratia apetala Buch.-Ham led to the separation of the new sesquiterpene, acorenone C (375), which exhibited moderate activity against AChE with a 23.34% inhibition ratio at 50 μM [A study on at 50 μM .Nemania bipapillata (AT-05) from the marine red alga Asparagopsis taxiformis-Falkenbergia stage led to the discovery of (+)- (376), (+)--4-deacetyl-5-hydroxy-botryenalol (377), (+)--4-deacetyl-botryenalol (378), (+)- (379), (+)--(380), and 4β-acetoxy-9β,10β,15α-trihydroxyprobotrydial (381). Compounds 376–381 showed AChE and BChE inhibition with inhibitory ratios of 18.3% and 27.7%, and 3.2% and 7.3% at 100 μM, respectively [Comprehensive research on ectively .382) was identified in Xylaria sp. HNWSW-2. The inhibition rate of 382 against AChE was 12.9% at 50 µg/mL [Guaidiol (50 µg/mL .383) was collected from a co-culture of Nigrospora oryzae and Irpex lacteus. This compound showed an AChE inhibitory capacity with a ratio of 35% at 50 µM [Nigrosirpexin A , which inhibited AChE with IC50 at 28 μg/mL [A chemical assay for 28 μg/mL .Armillaria sp. and endophyte Epicoccum sp. generated five protoilludane-type sesquiterpenoids, epicoterpenes A−E (385–389), which were inactive for AChE inhibition at 50 μM [A co-culture of at 50 μM .Phomopsis sp. TJ507A from Phyllanthus glaucus led to the identification of a 2,3-seco-protoilludane-type sesquiterpene, phomophyllin A (390); eight protoilludane-type sesquiterpenes, phomophyllins B−I (391–398); four illudalane-type sesquiterpenes, phomophyllins J−M ; and a botryane-type sesquiterpene, phomophyllin N (403). In addition, seven known sesquiterpenoids, granulone B (404), radulone B (405), 2-ethanol (406), pterosin Z (407), onitin (408), dehydrobotrydienol (409), and 7-hydroxy-10-oxodehydrodihydrobotrydial (410), were also isolated from this fungus. This represents the first natural product of 390 with an irregular 2,3-seco-protoilludane skeleton. Compounds 390–396, 398, 405, 408, and 410 inhibited BACE1 within the range of 19.4% to 43.8% at 40 μM [The comprehensive chemical investigation of at 40 μM .Colletotrichum gloeosporioides GT-7 generated the compound colletotrichine B (411), which inhibited AChE with IC50 at 38.0 ± 2.67 μg/mL [The fungus 67 μg/mL .Colletotrichum sp. SCSIO KcB3-2 from Kandelia candel produced a new polychiral bisabolane sesquiterpene of bisabolanoic acid A (412), which exhibited mild AChE inhibition with an IC50 of 2.2 μM, whereas the IC50 for the positive control of huperzine A was 0.30 ± 0.06 μM [A chemical assay for  0.06 μM .Penicillium sp. SK5GW1L, a mangrove endophytic fungus from the leaves of Kandelia candel, resulted in the separation of two new α-pyrone meroterpenoids, arigsugacin I (413) and 3-epiarigsugacin E (416), together with seven known analogs: arigsugacin F (414), territrem B (415), arisugacin D (417), arisugacin B (418), territrem C (419), and terreulactone C (420)  . The IC5ectively ,93.Aspergillus terreus Thom, an endophytic fungus from Tripterygium wilfordii Hook. f. (Celastraceae), revealed six undescribed meroterpenoids, spiroterreusnoids A–F (421–426). The IC50s of 421–426 for BACE1 and AChE inhibition ranged from 5.86 to 27.16 μM and from 22.18 to 32.51 μM, respectively [The investigation of ectively .Aspergillus 16-5c, a mangrove endophytic fungus identified from Sonneratia apetala, found one new meroterpenoid, 2-hydro-acetoxydehydroaustin (427), along with known analogs 11-acetoxyisoaustinone (428), isoaustinol (429), austin (430), austinol (431), acetoxydehydroaustin (432), dehydroaustin (433), dehydroaustinol (434), preaustinoid A2 (435), and 1,2-dihydro-acetoxydehydroaustin B (436). The IC50s for AChE inhibition by compounds 429, 433, and 434 were 2.50, 0.40, and 3.00 µM, respectively [A detailed study on ectively .Penicillium chrysogenum MT-12 collected from Huperzia serrata revealed the new compounds penicichrysogene A (437) and penicichryso-gene B (438)  . UnfortuAspergillus sp. YXf3 of Ginkgo biloba found an irregular C18 norditerpenoid, aspergiloid I (439), which did not exhibit antioxidant properties or AChE inhibition at 50 μg/mL [A study on the 50 μg/mL .440), along with four known steroids, containing -ergosta-4,6,8(14),22-tetraen-3-one (441), -3β-hydroxyergosta-5,8,22-trien-7-one (442), -ergosta-7,22-dien-3β,5α,6β-triol (443), and -5α,8α-epidioxyergosta-6,22-dien-3β-ol (444)  , were id00 µg/mL .Aspergillus flavus cf-5 from the marine red alga Corallina officinalis led to the separation of a new compound, 3β,4α-dihydroxy26-methoxyergosta-7,24(28)-dien-6-one (445), as well as four known isolates: episterol (446), -ergosta7,22-dien-3β,5α,6α-triol (447), -ergosta-5,22-dien-3β-ol (448), and -ergosta-4,6,8(14),22-tetraen-3-one (441). Compound 445 displayed weak activity against AChE with an inhibition ratio of 5.5% at 100 µg/mL [The instentive investigation of 00 µg/mL .Chaetomium sp. M453 associated with Huperzia serrata (Thunb. ex Murry) Trev produced the isolation of neocyclocitrinols E–G (449–451) and 3β-hydroxy-5,9-epoxy--ergosta-7,22-dien-6-one (452) as well as three known steroids (453–455) separated from the endophytic fungus Chaetomium sp. M453 associated with Huperzia serrata (Thunb. ex Murry) Trev. Compounds 451–452 were assayed for AChE inhibitory activities. Compound 452 alone showed weak AChE inhibitory activity at 50 μM [A study on at 50 μM .E)-3-hydroxy-5,6-epoxy7-one-8(14),22-dien-ergosta (456), 443, β-sitostenone (457), and β-sitosterol (458), and 448 were obtained from Chaetomium sp. YMF432 related to Huperzia serrata (Thunb. ex Murray) Trev. Compound 456 alone showed moderate AChE inhibition with an IC50 of 67.8 ± 1.7 μM and an inhibitory rate of 58.8 % at 100 μg/mL [Four known steroids,  from a branch of Bruguiera gymnoihiza (Linn.) Savigny resulted in the separation of 3β,5α-dihydroxy--ergosta-7,22-dien-6-one (459), 3β,5α,14α-trihydroxy--ergosta-7, 22-dien-6-one (460), and NGA0187 (461). Compound 461 displayed remarkable anti-AChE activity with an IC50 value of 1.89 μM [An extensive study on  1.89 μM .462) was identified from Curvularia sp. associated with Rauwolfia macsrophylla. The IC50 of 462 for AChE inhibitory activity was 1.52 μM [Ergosterol ( 1.52 μM .441 and (17R)-4-hydroxy-17-methylincisterol (463), were identified from Alternaria alternate related to the leaves of Psidium littorale Raddi. Compounds 441 and 463 were inactive for neuroprotective activity toward glutamate-injured PC12 cells at 40 and 80 μM, respectively [Two known steroids, ectively .Colletotrichum sp. F168 from the plant Huperzia serrata Trev produced the compound ergosta-7,22-dien-5,9-epoxy--6-one-3-yl acetate (464), which showed a negligible AChE inhibitory activity of 18.2% at 100 µg/mL [Research on 00 µg/mL .Talaromyces sp. SCNU-F0041 from the fresh leaves of Kandelia produced cyclosecosteroid A (465), ergosterol (462), -5α,8α-epidioxyergosta-6,22-dien-3β-ol (466), and cerevisterol (443). The IC50 of compound 465 for inhibiting AChE was 46 μM [The investigation of as 46 μM .448), 5,6-epoxyergosterol (454), citreoanthrasteroid A (467), demethylincisterol A (463), and chaxine C (468) were identified in Phyllosticta capitalensis derived from the leaves of Loropetalum chinense var. rubrum. Compound 467 alone exhibited neuroprotection with an EC50 of 24.2 μM for glutamate-mediated PC12 cell injury [Brassicasterol (l injury .Aspergillus (13), Penicillium (11), Colletotrichum (9), Phomopsis (5), Talaromyces (4), Chaetomium (4), Alternaria (2), Epicoccum (2), Cochliobolus (2), and Curvularia (2)  . Around 17) and isochaetoglobosin D (23), which showed strong H2O2-induced PC12 cell damage-inhibiting activities with EC50s of 0.003 ± 0.0003 and 0.009 ± 0.001 μM, respectively. Huptremules C and D  showed stronger AChE-inhibiting activities, with IC50s of 0.11 ± 0.01 and 0.06 ± 0.00 μM, respectively, than hupA (IC50 = 0.54 μM). Hence, they represent valuable compounds for developing anti-AD agents. Notably, structural changes to these compounds directly affect their bioactivities. Synthesis and structural modifications for bioactive metabolites are necessary to prepare more effective analogs. This review confirmed the significance of endophytes in the generation of abundant metabolic products with anti-AD activities. In the future, with the addition of further in-depth research on endophytic fungal metabolites, more biologically active chemical resources will become available to medicinal chemists and biologists for anti-AD drug research.Based on the analyzed data, the biological activity of these compounds was determined, mainly focusing on their anti-AChE, anti-BChE, antioxidant, and neurotrophic activities. Some of the compounds exhibited micromolar to nanomolar biological activities, such as chaetoglobosin F (


Dacryodes edulis is a plant that belongs to the Burseraceae family. It is widely used traditionally alone or in association with other plants in Cameroonian folk medicine to cure wounds, fever, headaches, and malaria. The aim of this work was to investigate the leaves and stem bark of D. edulis with an emphasis on the antiplasmodial and cytotoxic effects of extracts, fractions, and isolated compounds.Plasmodium falciparum using a SyBr Green fluorescence-based assay. The cytotoxicity of active extracts, fractions, and compounds was tested against mammalian Raw cell lines using an in vitro resazurin-based viability assay. The structures of the compounds were determined based on their NMR and MS data. The in vivo toxicity using female BALB/c mice was performed on the most active extract according to the protocol of OECD (2002), guideline 423.Extracts, fractions, and some isolated compounds were subjected to antiplasmodial activity screening in vitro against chloroquine-sensitive 3D7 and multidrug resistant Dd2 strains of D. edulis displayed good antiplasmodial activity with IC50 values of 3.10 and 3.56 μg/mL respectively on sensitive (3D7) and multiresistant (Dd2) strains of P. falciparum. Of the sixteen compounds isolated, 3,3′,4-tri-O-methylellagic acid (4) exhibited the highest antiplasmodial activity against PfDd2 strains with an IC50 value of 0.63 μg/mL. All extracts, fractions, and isolated compounds demonstrated no cytotoxicity against Raw cell lines with CC50 > 250 μg/mL. In addition, the most active extract on both strains of P. falciparum was nontoxic in vivo, with a LD50 greater than 2000 and 5000 mg/kg. A phytochemical investigation of the stem bark and leaves of D. edulis afforded sixteen compounds, including two xanthones (1–2), three ellagic acid derivatives (3–5), one phenolic compound (6), one depside (7), one triglyceride (8), one auranthiamide acetate (9), one gallic acid derivative (10), four triterpenoids (11–14), and two steroids (15–16). Compounds 1, 2, 5, 7, 8, and 9 were herein reported for the first time from the Burseraceae family.The hydroethanolic extract from the leaves of O-methylellagic acid and ethylgallate) from the plant. These biological results support the use of D. edulis in traditional medicine against malaria.This work highlights the good in vitro antiplasmodial potency of the hydroethanolic extract of the leaves of this plant and that of two isolated constituents (3,3′,4-tri-The online version contains supplementary material available at 10.1186/s12906-023-03957-2. Plasmodium falciparum, malaria continues to decimate the population mostly in Africa. Faced with these health issues, the search for new active, safer and more efficient antimalarials with a novel mode of action to combat pathogenic agents and of almost zero toxicity is becoming imperative. To find such agents, medicinal plant-based drug discovery remains a hard-wearing solution. As such, medicinal plants always have a strong historical link with malaria treatment, exemplified by quinine and artemisinin isolated from Cinchona trees and Artemisia annua, respectively. Hence, investigating plants widely used in traditional medicine to treat malaria can provide new drugs with an innovative biological mode of action. Dacryodes edulis (G. Don) HJ. Lam. (Burseraceae) is traditionally used alone or in association with other plants in folk medicine to cure wounds, fever, headaches, and malaria , β-and α- amyrin (14) , β-amyrin (12)  and the mixture of β-sitosterol and stigmasterol (15) . The ethyl acetate fraction was assessed by CC over silica gel  and eluted with a DCM/MeOH mixture (1:0–1:0) of increasing polarity to afford glyceryl-1-tetracosanoate (8), auranthiamide acetate (9) (4.9 mg), and ethyl gallate (10). The structures of these compounds  of er 2.5 × 0 L  . The colPf3D7-(MRA-102) and chloroquine-resistant PfDd2-(MRA-150) P. falciparum strains were maintained as previously described  and incubating at 37 °C in a humidified atmosphere with 5% CO2. The medium was renewed daily to propagate the culture. Giemsa-stained thin blood smears were examined microscopically under immersion oil to monitor cell-cycle transition and parasitemia evolution. O+ human red blood cells were obtained from a consent volunteer donor (The chloroquine-sensitive escribed  in fresher donor .Whole blood (25 mL) was drawn for a heathy volunteer without a historical contraceptive or anticoagulant therapy using a protocol approved by the Institutional Review Board (IRB No. 001/UY11 BTC/IRBI 2009), Biotechnology Centre, University of Yaoundé I, Cameroon. A consent form was approved by Fekam Boyom Fabrice, Professor and Head of the Antimicrobial & Biocontrol Agents Unit (AmBcAU), Laboratory for Phytobiochemistry and Medicinal Plants Studies, University of Yaoundé 1, Cameroon, for collection of blood sample from human volunteer.The volunteer donor was supplied a consent form which informed the purpose of the research project, name and details as well as contact person. Inclusion and exclusion criteria of the donor include volume of blood to be taken, age, sex, no medical illness, no medication, and time required for the blood sampling. Information’s were asked during face to face interview. Blood donor was aware that his sample might be used for research purpose and written consent form was obtained.P. falciparum in culture.Before each experiment, synchronized ring-stage parasite were obtained by 5% sorbitol (w/v) treatment with respect to Lambros and Vanderberg, 1979 . It is cA drug sensitivity assay was carried out in 96-well microtitration plates using a SYBR green I-based fluorescence assay . This asμL) followed by incubation at 37℃ for 72 h. After incubation, 100 μL of SYBR Green I buffer  was added to each well, mixed twice gently with a multichannel pipette and incubated in the dark at 37℃ for 1 h +, . 1H-NMR : 2.91 , 3.78 , 3.81 , 6.55 , 6.60 , 6.79 , 6.86 , 13.91 . 13C-NMR : 23.8 (C-15), 56.2 (C-16), 56.2 (C-14), 92.8 (C-4), 97.8 (C-2), 99.5 (C-5), 104.7 (C-10), 113.4 (C-13), 116.3 (C-7), 143.8 (C-8), 157.7 (C-11), 160.0 (C-12), 164.7 (C-6), 164.8 (C-1), 166.8 (C-3), 183.0 (C-9) +, .1H-NMR : 13.39 , 6.68 , 6.66 , 6.35 , 6.31 , 3.86 , 2.88 . 13C-NMR : 182.6 (C-9), 165.9 (C-3), 163.8 (C-1), 163.7 (C-6), 159.4 (C-11), 156.9 (C-12), 143.5(C-8), 115.6 (C-7), 113.0 (C-13), 104.2 (C-10), 98.5 (C-5), 96.8 (C-2), 92.1 (C-4), 55.7 (C-14), 23.6 (C-15) +, . 1H-NMR : 3.59 , 4.17 , 4.99 , 8.04  +. . 1H-NMR : 3.85 , 4.13 , 4.19 , 7.82 , 8.03 . 13C-NMR : 56.4 (-OCH3), 61.1 (-OCH3), 61.3 (-OCH3), 107.8 (C-5′), 111.6 (C-1′), 112.6 (C-1), 112.9 (C-5), 113.6 (C-6′), 114.1 (C-6), 141.1 (C-2), 142.1 (C-3′), 141.8 (C-2′), 142.1 (C-3), 154.1 (C-4′), 154.3 (C-4), 158.9 (C-7′), 159.0 (C-7) +, . 1H-NMR : 8.66 , 8.36 , 7.85 , 6.20 , 5.91 , 4.73 , 4.53 , 4.46 , 4.38 , 4.88 , 4.87 . 13C-NMR : 174.9 (C-7‴), 168.0 (C-7), 167.9 (C-7′), 162.5 (C-4′), 160.5 (C-4), 155.0 (C-3‴/5‴), 151.5 (C-6), 151.2 (C-6′), 150.5 (C-5), 149.7 (C-5′), 147.7 (C-4‴), 129.4 (C-1‴), 124.1 (C-2), 122.4 (C-2′), 122.6 (C-1′), 121.5 (C-1), 121.2 (C-3), 120.6 (C-3′), 118.3 (C-2‴), 118.3 (C-6‴), 110.9 (C-1″), 86.7 (C-3″), 80.7 (C-2″), 76.9 (C-4″), 75.2 (C-5″), 71.2 (OCH3), 70.5 (OCH3)  +, . 1H NMR : 11.36 (OH), 6.62 , 6.56 , 6.49 , 6.32 , 4.10 , 2.75 , 2.44 , 1.65 , 1.55 , 1.36 , 1.22 , 0.91 , 0.85 . 13C-NMR : 207.1 (C-9), 170.5 (C-7′′), 169.2 (C-7), 166.6 (C-2), 164.9 (C-4), 157,8 (C-2′′), 151.4 (C-4′′), 144.5 (C-6′′), 138.9 (C-6), 119.9 (C-1′′), 115.2 (C-5′′), 113.4 (C-5), 104.2 (C-1), 103.1 (C-3′′), 100.1 (C-3), 56.4 (-OCH3), 55.5 (-OCH3), 51.2 (C-8), 42.6 (C-10), 33.8 (C-8′′), 31.6 (C-10′′), 31.3 (C-12), 30.7 (C-9′′), 23.4 (C-11), 22.5 (C-11′′), 22.4 (C-13), 14.0 (C-14), 13.9 (C-12′′) +.1H-NMR : 4.21 , 4.15 , 3.93 , 3.69 , 3.60 , 2.35 , 1.63 , 1.25 , 0.88 . 13C-NMR : 174.4 (C-1ʹ), 70.2 (C-2), 65.2 (C-1), 63.6 (C-3), 34.3 (C-2ʹ), 24.9 (C-3ʹ), 29.7- 32.0 (C4ʹ-23ʹ), 14.1 (C-24ʹ) +, . 1H-NMR : 7.71 , 7.52 , 7.44 , 7.25 , 7.17 , 6.71 , 5.90 , 3.93 , 3.82 , 3.22 , 3.05 . 13C-NMR : 170.7 (C-2), 170.2 (C-6), 167.1 (C-9), 136.7 (C-1ʹ), 136.6 (C-1″), 132.0 (C-1‴), 129.3 (C-4‴), 129.1 (C-4ʹ), 129.1 (C-3‴/5‴), 128.7 (C-2‴/6‴), 128.6 (C-3″/5″), 128.5 (C-2ʹ/6′), 127.1 (C-2″/6″), 127.0 (C-3ʹ/5′), 126.7 (C-4″), 64.5 (C-3), 55.1 (C-7), 49.4 (C-4), 38.3 (C-11), 37.4 (C-10), 20.8 (C-1ʺ) +. 1H-NMR : 6.93 , 5.67 , 2.52 , 0.93 , 0.83 . 13C-NMR : 219.1 (C-3), 173.2 (C-26), 148.9 (C-8), 145.5 (C-24), 126.6 (C-25), 121.2 (C-7), 52.9 (C-17), 52.3 (C-5), 52.0 (C-14), 47.0 (C-4), 45.4 (C-9), 44.0 (C-13), 34.2 (C-2), 18.2 (C-27) [Masticadienonic acid or 3-oxo-lanosta-7,24- 126.6 C-, 121.2  and the multidrug-resistant (Dd2) strains of P. falciparum was evaluated by measuring growth inhibition based on SYBR green fluorescence. Chloroquine (CQ) and artemisinin were used as reference compounds. To classify the antiplasmodial activity expressed as IC50 of tested samples, the following criteria were adopted: IC50 ≤ 5 μg/mL: pronounced activity; 5 < IC50 ≤ 10 μg/mL: good activity; 10 < IC50 ≤ 20 μg/mL: moderate activity; 20 < IC50 ≤ 40 μg/mL: low activity; IC50 > 40 μg/mL: inactive [D. edulis inhibited the growth of both the P. falciparum 3D7 and Dd2 strains with IC50 values ranging from 1.44 to 23.39 μg/mL  . Overall, we can observe that fractionation of both extracts led to some fractions with more potent and pronounced activities than the mother extracts. This observation could be explained by the fact that the phytochemical constituents responsible for the antiplasmodial activity in the mother extract were concentrated in potent fractions during the fractionation process. Of note, fractionation of an extract can positively or negatively change its biological properties by concentrating active ingredients into a fraction, or by sharing them between the various fractions [O-dimethylellagic acid (3) and ethylgallate (10) displayed pronounced antiplasmodial activities on both Pf3D7 and PfDd2. Interestingly, 3-oxo-7,24-Z-tirucalladien-26-oic acid (13); a (1:1) mixture of β- and α- amyrin (14), 3,3′,4-tri-O-methylellagic acid (4) and 3,4-dihydroxybenzoic acid (6) exhibited pronounced to moderate antiplasmodial activity only on the multidrug-resistant Dd2 strain of P. falciparum with IC50 values ranging from 0.63 to 17.09 μg/mL. However, 3, 3′, 4-tri-O-methylellagic acid (4) and the mixture of α- and β- amyrin (14) were the most potent compounds with respective IC50 values of 3.14 and 0.63 μg/mL, whereas 3,4-dihydroxybenzoic acid (6) and 3-oxo-lanosta-7,24-Z-dien-26-oid acid (13) displayed moderate antiplasmodial activity  (3) and the other from the hydroethanolic leaf extract  (10) with very good antiplasmodial potency. These isolates could be interesting starting materials for further structure–activity relationship studies. The results summary is shown in Table Among fractions from the hydroethanol leaf extract, fraction DFH was the only one showing pronounced antiplasmodial activity on both strains  values > 250 μg/mL for the tested extract and all the fractions and > 100 μg/mL for the isolated compounds. Futhermore, the hydroethanolic extract of D. edulis leaf extract at the tested limit doses of 2000 and 5000 mg/kg did not cause any death during the 14 days of experimentation, indicating that the median lethal dose (LD50) of this extract is greater than 5000 mg/kg. Therefore, according to the OECD’s Globally Harmonized System of Classification [D. edulis leaf extract can be classified as category 5 and considered nontoxic once administered orally. Additionally, the recording of body weight during 14 days of observation showed no significant change in treated animals compared to the normal group  who found significant activity against both chloroquine-sensitive (3D7) and chloroquine-resistant (Dd2) strains of P. falciparum with IC50 values of 4.34 and 6.43 μg/mL, respectively [Bursera simaruba (Burseraceae) from Colombian North Coast with IC50 values of 1.7 and 1.2 μg/mL on the chloroquine sensitive (3D7) and chloroquine-resistant (Dd2) strains, respectively [D. edulis constitutes a powerful source of antiplasmodial compounds. Nevertheless, in the work conducted by Zofou and colleagues, fractionation of methylene chloride/methanol extract led to the isolation of quercitrin, afzelin, quercetin, methyl 3,4,5-trihydroxybenzoate, and sitosterol 3-O-β-D-glucopyranoside sterol with antiplasmodial activities ranging from 0.37 to 18.53 μg/mL against both sensitive Pf3D7 and resistant PfDd2 strains of P. falciparum. This discrepancy in activities could be explained by the different extraction procedures, the part of plant used, which is directly linked to the quantity and quality of secondary metabolites present in the crude extract [50: 17.09 μg/mL) on multi-resistant (Dd2) strain of P. falciparum compared to methyl 3,4,5-trihydroxybenzoate (DES4) (IC50: 0.56 μg/mL) isolated from the work achieved by Zofou and co-workers in 2013. The activity obtained by Zofou et al. is 30.51-fold more potent than the one obtained in our study. This discrepancy could be explained by the hydroxyl group at position 5 in methyl 3,4,5-trihydroxybenzoate absent in methyl 3,4-trihydroxybenzoate. From our investigation, 3,3'-di-O-methylellagic acid (3), 3,3″-di-O-methylellagic acid 4-O--β-D-xylopyranoside (5), Ethyl gallate (10) 3,3′,4-tri-O-methylellagic acid (4) belonging to the class of phenolics compound and derivative have been pointing out as the most highly potent antiplasmodial agents from D. Eludis. Phenolics compounds and derivative have been reported to displayed strong antiplasmodial potency. Of note, phenolic compounds are frequent in plants and are distributed within numerous phytochemical classes. They are broadly defined by their molecular structure, with one or more aromatic rings, without a nitrogen, originated from plantʼs metabolism pathways of the shikimate and/or acetate [P. falciparum by interfering with DNA topoisomerase, the induction of cell cycle arrest, and the activation of apoptotic pathways [Plasmodium parasite and an impairment of beta-hematin formation [P. falciparum have not yet been elucidated, reported data suggests that the hydroxy groups of the gallate moiety could play a pivotal role as donors in the establishment of bonds where the compound would exercise its activity [D. eludis is an inexhaustible source of drug candidate with antimalarial properties.The antiplasmodial activity of the methylene chloride/methanol extract of ectively . In 2022ectively . However extract . Besidespathways . Besidesormation –33. AlthD. edulis extracts from the leaves and the stem bark as well as their fractions and isolated compounds. The hydroethanolic leaves displayed better antiplasmodial activity than the methanolic extract of the stem bark, with IC50 values of 3.10 and 3.56 μg/mL on 3D7 and Dd2, respectively. Among the sixteen compounds isolated, 3,3′,4-tri-O-methylellagic acid exhibited the highest antiplasmodial activity against PfDd2 strains with an IC50 value of 0.63 μg/mL. All extracts, fractions, and isolated compounds demonstrated cytotoxicity against Raw cell lines with CC50 > 250 μg/mL. Moreover, the most active extract on both strains of P. falciparum was nontoxic in vivo with an LD50 greater than 5000 mg/kg. The results obtained in the present work highlighted safety and the antiplasmodial potency of the hydroethanolic extract of the leaves of this plant as well as the antiplasmodial activity of 3,3′,4-tri-O-methylellagic acid and ethylgallate. These biological results justify the use of D. edulis in traditional medicine against malaria, and can therefore contribute to hit optimization studies in antimalarial drug development .The objective of this work were to investigate the antiplasmodial activities and the toxicity of Additional file 1: Figure S1. HR-ESI mass spectrum of 1. Figure S2. 1H NMR spectrum of 1. Figure S3. 13C NMR spectrum  of 1. Figure S4. ESI mass spectrum of2. Figure S5. 1H NMR spectrum  of 2. Figure S6. 13C NMR spectrum  of 2. Figure S7. 1H NMR spectrum of 3. Figure S8. 1H NMR spectrum of 4. Figure S9. 13C NMR spectrum of 4. Figure S10. HR-ESI mass spectrum of 5. Figure S11. 1H NMRspectrum  of 5. Figure S12. 13C NMRspectrum  of 5. Figure S13. 1H NMRspectrum  of 6. Figure S14. 13C NMRspectrum  of 6. Figure S15. HR-ESI mass spectrum of 7. Figure S16. 1H NMRspectrum  of 7. Figure S17. 13C NMRspectrum  of 7. Figure S18. 1H NMRspectrum  of 8. Figure S19. 13C NMRspectrum  of 8. Figure S20. HR-ESI mass spectrum of 9. Figure S21. 1H NMRspectrum  of 9. Figure S22. 13C NMRspectrum  of 9. Figure S23. 1H NMRspectrum  of 10. Figure S24. 1H NMRspectrum  of 11. Figure S25. 13C NMRspectrum  of 11. Figure S26. 1H NMRspectrum  of 12. Figure S27. 13C NMRspectrum  of 12. Figure S28. 1H NMRspectrum  of 13. Figure S29. 13C NMRspectrum  of 13. Figure S30. 1H NMRspectrum  of 14. Figure S31. 13C NMRspectrum  of 14.


The structures of new compounds were established by 1D-, 2D-NMR and HR-ESI-MS spectral analyses. Compound 1 represents the second example of diarylbenzophenone from natural sources. Compound 2 possesses an unusual biphenyl-bisbenzophenone structure. Their cytotoxicity against human hepatocellular carcinoma HepG2 and SMCC-7721 cells and inhibitory activities on lipopolysaccharide-induced nitric oxide (NO) production in RAW264.7 cells were evaluated. Compound 2 showed moderate inhibitory activity against HepG2 and SMCC-7721 cells, and compounds 4 and 5 showed moderate inhibitory activity to HepG2 cells. Compounds 2 and 5 also exhibited inhibitory activities on lipopolysaccharide-induced nitric oxide (NO) production.Six compounds including three new benzophenones, selagibenzophenones D-F ( Selaginella (Selaginellaceae) comprises about 700 species and about 70 species were widely dispersed across the area south of the Yangtze River + quasi-molecular ion peak at 399.1243  in the HR–ESI–MS, which indicated 17 unsaturations. This molecular formula was consistent with the 1H and 13C NMR data , 6.59 , δ 7.02 , 6.58 , and δ 6.96 , 6.60 , and one orthotetra-substituted phenyls at δ 7.20 , 7.05  on the basis of 1H-1H COSY spectrum. The 13C NMR spectrum of 1 (δ 198.7 (C-7). According to these spectroscopic data compared with our reported selagibenzophenone C + quasi-molecular ion peak at 335.0918  in the HR–ESI–MS spectrum. The 1H NMR spectrum of 3 , 6.76  and δ 7.59 , 6.78 , and one 1,2,4-trisubstituted phenyls at δ 7.53 , 7.01 , 6.92 , which was confirmed by the 1H-1H COSY spectrum. The 13C NMR spectrum of 3 (δ 195.3 (C-7) and 196.5 (C-14). In HMBC spectrum, the correlations of H-3, 5 to C-7 and H-12 to C-7 indicated that ring A and ring B were connected with C-7. The HMBC couplings of H-9, 11 to C-14 and H-15, 19 to C-14 indicated that ring B and ring C were linked with C-14. In addition, the HMBC couplings of H-9, 12 to C-8 defined the location of a hydroxyl at C-8. Thus, the structure of compound 3 was elucidated and given a successive name, selagibenzophenone F, and its 3D structure was shown in Compound rum of 3  exhibiterum of 3  showed 2The phytochemical study resulted in the isolation of three new benzophenones (ariscina . The strcina (4) , selagin (4) + 399.1243; found 399.1232.Selagibenzophenone D (2): Yellow powder. UV (MeOH) λmax : 264 (4.26). 1H NMR and 13C NMR (MeOH-d4) see 34H25O9 [M + H]+ 577.1516; found 577.1499.Selagibenzophenone E (3): Yellow powder. UV (MeOH) λmax : 290 (4.22). 1H NMR and 13C NMR (MeOH-d4) see 20H15O5 [M + H]+ 335.0918; found 335.0919.Selagibenzophenone F (4): Light yellow amorphous powder. UV (MeOH) λmax (nm): 226, 264, 334. 1H NMR : δH 7.86 , 7.58 , 6.83 , 6.64 , 6.51 , 6.47 , 5.20 . 13C NMR : δC 170.6 (C-27), 157.4 , 156.6 (C-22), 151.1 (C-19), 141.5 (C-15), 138.1 (C-18), 137.5 (C-17), 130.5 , 130.2 , 129.9 , 129.8 (C-13), 129.5 (C-16), 129.4 (C-25), 122.0 (C-14), 114.1 , 93.7 (C-7), 59.3 (C-26).Selaginellin H (5): Yellow powder. UV (MeOH) λmax (nm): 280. 1H NMR : δH 7.70 , 7.64 , 7.42 , 7.11 , 6.98 , 6.78 , 6.67 , 6.66 , 4.89 . 13C NMR : δC 197.9 (C-7), 162.9 (C-1), 158.0 (C-24), 156.7 (C-16), 141.3 (C-9), 141.0 (C-13), 138.9 (C-12), 132.4 , 132.2 , 130.8 (C-19), 129.8 , 129.3 (C-11), 129.1 (C-4), 126.8 (C-10), 119.0 (C-8), 115.0 , 114.9 , 114.6 , 113.2 (C-27), 99.2 (C-21), 82.5 (C-20), 61.7 (C-28).Selaginellin S (6): Light yellow amorphous powder. UV (MeOH) λmax (nm): 226, 264, 324. 1H NMR : δH 8.00 , 7.98 , 7.54 , 7.02 , 6.77 , 6.62 , 6.37 . 13C NMR : δC 182.9 (C-4), 165.6 (C-7″), 164.7 (C-2), 162.7 (C-7), 161.2 (C-2″), 160.8 (C-5), 156.1 (C-1″), 159.1 (C-9), 134.8 (C-6″), 130.8 (C-4″), 128.0 , 121.9 (C-5″), 121.8 (C-1′), 118.8 (C-4′), 115.4 , 114.8 (C-3″), 104.9 (C-8), 103.9 (C-10), 101.8 (C-3), 98.8 (C-6).Unciflavone D , with sorafenib as a positive control. Three parallel holes were located and then incubated for 48 h. Subsequently, the 96 well plate was taken out and 10 µL of CCK-8 was added in DMEM 0.1 mL; meanwhile, two separate holes were used as a blank control, with only 10 µL CCK-8 in DMEM 0.1 mL added to each blank hole. Then it was incubated under the same conditions for 4 h. The optical density (OD) was measured at 450 nm using a Biotek Synergy . The experiment was repeated 3 times. The IC50 values were calculated to assure the impact of the drugs on cell growth inhibition rate.The human hepatocellular carcinoma HepG2 and SMCC-7721 cell line was purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences of Chinese Academy of Sciences. The cytotoxicity assay was carried out using the Cell Counting Kit-8 (CCK-8) method. HepG2 and the SMCC-7721 cell line were cultured in DMEM at 37 °C, 5% CO1–6 by inhibiting NO release in LPS-induced murine macrophage RAW 264.7 cells. The RAW 264.7 cell line was purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences of Chinese Academy of Sciences. The cells were cultured in an incubator and DMEM at 37 °C in 5% CO2. The cells were seeded in 24-well culture plates  and allowed to adhere for 24 h at 37 °C. A blank control group and a drug group were cultured for 2 h in an incubator. 10 μg/mL of LPS  per well was added to induce inflammation, and was cultured in the incubator for 24 h. As a parameter of NO synthesis, the nitrite concentration was measured by the Griess reaction using the supernatant of the RAW 264.7 cells. The absorbance was read with a microplate reader  at 540 nm. The experiment was performed three times. SPSS 16.0 and GraphPad Prism 6.01 software were used for statistical analysis.NO represents an important inflammatory factor. In this study, we examined the NO inhibitory effects of compounds 1–3), two known selaginellins and one known flavonoid were isolated from Selaginella tamariscina. The structures of the new compounds were elucidated by spectroscopic analysis. Compound 1 represents the second example of a diarylbenzophenone from natural sources. Compound 2 possesses an unusual biphenyl-bisbenzophenone structure. Compounds 1–6 from this plant were evaluated for their cytotoxicity against human hepatocellular carcinoma HepG2 cells and human SMCC-7721 cells. Compound 2 showed moderate inhibitory activity against HepG2 and SMCC-7721 cells with IC50 values of 32. 575 and 15.816 µM, respectively. Compounds 4 and 5 exhibited moderate inhibitory activity against HepG2 cells. In addition, all compounds were evaluated for their inhibitory activities on lipopolysaccharide-induced nitric oxide (NO) production in RAW264.7 cells. Compounds 2 and 5 exhibited inhibitory activities on lipopolysaccharide-induced nitric oxide (NO) production.In this study, three new benzophenone derivatives, selagibenzophenones D-F (


All complexes have been characterized using different spectroscopic methods, viz. IR, UV-Vis, circular dichroism, one-  and two-dimensional  NMR spectroscopy, and elemental analysis. Furthermore, the catalytic ability of all compounds in the epoxidation of styrene, cyclohexene, and its naturally occurring monoterpene derivatives, i.e., S(−)-limonene and (−)-α-pinene has also been studied, using two different oxidants, i.e., aqueous 30% H2O2 or tert-butyl hydroperoxide (TBHP). In addition, the biological properties of these chiral oxidovanadium(V) compounds, but also cis-dioxidomolybdenum(VI) complexes with the same chiral Schiff bases, were studied. Their cytotoxic and cytoprotective activity studies with the HT-22 hippocampal neuronal cells revealed a concentration-dependent effect in the range of 10–100 μM. Moreover, vanadium(V) complexes, in contrast to cis-dioxidomolybdenum(VI) compounds, demonstrated higher cytotoxicity and lack of cytoprotective ability against H2O2-induced cytotoxicity.New oxidovanadium(V) complexes,  Amanita muscaria , [VO2], [VO2] [2(acac-aebmz)], and conversions from 51 to 78%. Unexpectedly, when TBHP was used as the terminal oxidant with catalytic amounts of these complexes, in contrast to 1–10VOL catalysts, conversions significantly decreased to even 20%, but selectivity towards styrene oxide was slightly better, up to 47% in the case of [VO2(acac-ambmz)].On the other hand, as well as for reported earlier, [VO-aebmz)] , and [VO-aebmz)] , styrene1–10VOL complexes and with both oxygen sources, i.e., aqueous 30% H2O2 or tert-butyl hydroperoxide (TBHP). The reaction goes in two independent ways with cyclohexene oxide and, after its eventual hydrolysis, cyclohexene-1,2-diol as an epoxidation product, but also allylic oxidation products were observed, such as 2-cyclohexen-1-ol and after further oxidation resulting in the formation of 2-cyclohexen-1-one , but significantly lower selectivity towards epoxide (29%) and 2-cyclohexen-1-one (8%), but high amounts of cyclohexen-1-ol (55%) [2-OCH3)(L1)]2, [VO2(L2)]·H2O and [VO2(L3)]  . In contectivity .S(−)-limonene and (−)-α-pinene. In comparison to cyclohexene, S(−)-limonene was oxidized by 30% H2O2 with very low conversions (S(−)-limonene with both oxidants, but distinctly better selectivity was achieved towards (−)-α-pinene oxide (up to 73% with TBHP).Finally, the catalytic ability of all complexes was studied in the oxidation of mono- and bicyclic naturally occurring monoterpene derivatives of cyclohexene, giving analogous epoxidation and allylic oxidation products, i.e., versions , but epoversions  gave con3VOL, 5VOL and 6VOL, and for comparison cis-dioxidomolybdenum(VI) complexes with the same chiral tetradentate Schiff base ligands [2(HL3)][MoO and 2(HL6)][MoO and 75% for 2(HL6)]: 322 (9350), 445 (1230). CD spectrum in DMSO : 270 (−1.96), 309 (−1.32), 367 (2.69). 1H NMR : 8.69  (azomethine); 8.25 , 7.67 , 7.55 , 6.98  (aromatic); 5.99 , 4.09  (methine); 4.30 , 4.22  (methylene). 51V NMR : −541.9. 2VOL: Yield 86%. Anal. Calc. for C17H15N2O7V: C, 49.8; H, 3.7; N, 6.8. Found: C, 49.7; H, 3.6; N, 6.9%. IR : 1623 (νC=N); 1600 (νC=C); 1521, 1346 (νNO2); 1290, 1080 (νC-O); 978 (νV=O). UV-Vis spectrum in DMSO : 338 (4520), 452 (1140). CD spectrum in DMSO : 272 (−1.83), 318 (−1.24), 379 (2.39). 1H NMR : 8.67  (azomethine); 8.24 , 7.66 , 7.22 , 7.14 , 6.92  (aromatic); 5.97 , 4.08  (methine); 4.29 , 4.22  (methylene); 3.93  (methoxy). 51V NMR : −537.6.3VOL: Yield 92%. Anal. Calc. for C17H15N2O7V: C, 49.8; H, 3.7; N, 6.8. Found: C, 49.8; H, 3.8; N, 6.8%. IR : 1629 (νC=N); 1607 (νC=C); 1519, 1346 (νNO2); 1276, 1036 (νC-O); 965 (νV=O). UV-Vis spectrum in DMSO : 348 (4680), 477 (1190). CD spectrum in DMSO : 278 (−1.65), 322 (−1.13), 387 (2.24). 1H NMR : 8.76  (azomethine); 8.22 , 7.63 , 7.19 , 7.07 , 6.89  (aromatic); 5.97 , 4.07  (methine); 4.28 , 4.20  (methylene); 3.79  (methoxy). 51V NMR : −536.7.4VOL: Yield 90%. Anal. Calc. for C17H15N2O6V: C, 51.8; H, 3.8; N, 7.1. Found: C, 51.9; H, 3.7; N, 7.0%. IR : 1625 (νC=N); 1602 (νC=C); 1521, 1346 (νNO2); 1287, 1069 (νC-O); 982 (νV=O). UV-Vis spectrum in DMSO : 336 (5570), 454 (1250). CD spectrum in DMSO : 271 (−1.60), 309 (−1.10), 375 (2.26). 1H NMR : 8.62  (azomethine); 8.24 , 7.64 , 7.40 , 7.33 , 6.88  (aromatic); 5.94 , 4.08  (methine); 4.30 , 4.24  (methylene); 2.32  (methyl). 51V NMR : −539.0.5VOL: Yield 87%. Anal. Calc. for BrC16H12N2O6V: C, 41.9; H, 2.6; N, 6.1. Found: C, 41.8; H, 2.7; N, 6.0%. IR : 1626 (νC=N); 1604 (νC=C); 1521, 1346 (νNO2); 1283, 1072 (νC-O); 976 (νV=O). UV-Vis spectrum in DMSO : 340 (5420), 448 (1270). CD spectrum in DMSO : 277 (−1.31), 304 (−1.05), 373 (2.15). 1H NMR : 8.67  (azomethine); 8.26 , 7.73 , 7.71 , 7.61 , 6.90  (aromatic); 6.04 , 4.03  (methine); 4.24 , 4.10  (methylene). 51V NMR : −542.1.6VOL: Yield 81%. Anal. Calc. for C16H12N3O8V: C, 45.2; H, 2.8; N, 9.9. Found: C, 45.1; H, 2.9; N, 10.0%. IR : 1632 (νC=N); 1607 (νC=C); 1520, 1344 (νNO2); 1316, 1100 (νC-O); 972 (νV=O). UV-Vis spectrum in DMSO : 343 (8210), 426 (3400). CD spectrum in DMSO : 275 (−1.23), 311 (−0.94), 354 (1.89). 1H NMR : 8.85  (azomethine); 8.55 , 8.35 , 8.29 , 7.78 , 7.03  (aromatic); 6.24 , 3.96  (methine); 4.21 , 4.01  (methylene). 51V NMR : −540.4.7VOL: Yield 83%. Anal. Calc. for C16H13N2O7V: C, 48.5; H, 3.3; N, 7.1. Found: C, 48.5; H, 3.4; N, 7.1%. IR : 1630 (νC=N); 1606 (νC=C); 1542, 1350 (νNO2); 1293, 1109 (νC-O); 974 (νV=O). UV-Vis spectrum in DMSO : 354 (5830), 458 (1360). CD spectrum in DMSO : 270 (−1.69), 312 (−1.17), 367 (2.36). 1H NMR : 9.72  (hydroxyl); 8.46  (azomethine); 8.25 , 7.62 , 7.50 , 6.46 , 6.29  (aromatic); 5.91 , 4.07  (methine); 4.30 , 4.19  (methylene). 51V NMR : −537.5.8VOL: Yield 86%. Anal. Calc. for C20H21N2O6V: C, 55.1; H, 4.9; N, 6.4. Found: C, 55.0; H, 4.9; N, 6.3%. IR : 1624 (νC=N); 1591 (νC=C); 1521, 1346 (νNO2); 1289, 1084 (νC-O); 975 (νV=O). UV-Vis spectrum in DMSO : 349 (6670), 449 (1440). CD spectrum in DMSO : 263 (−1.37), 315 (−0.67), 371 (2.20). 1H NMR : 8.68  (azomethine); 8.24 , 7.66 , 7.60 , 7.41 , 6.94  (aromatic); 5.93 , 4.12  (methine); 4.31 , 4.25  (methylene); 1.51  (tert-butyl). 51V NMR : −552.7.9VOL: Yield 82%. Anal. Calc. for C24H29N2O6V: C, 58.5; H, 5.9; N, 5.7. Found: C, 58.7; H, 5.8; N, 5.7%. IR : 1620 (νC=N); 1598 (νC=C); 1521, 1346 (νNO2); 1296, 1080 (νC-O); 964 (νV=O). UV-Vis spectrum in DMSO : 351 (6320), 451 (1330). CD spectrum in DMSO : 265 (−1.36), 302 (−1.12), 374 (2.13). 1H NMR : 8.70  (azomethine); 8.26 , 7.67 , 7.49 , 7.38  (aromatic); 5.96 , 4.14  (methine); 4.29 , 4.23  (methylene); 1.53 , 1.41  (tert-butyl). 51V NMR : −547.3.10VOL: Yield 88%. Anal. Calc. for C20H15N2O6V: C, 55.8; H, 3.5; N, 6.5. Found: C, 55.9; H, 3.4; N, 6.6%. IR : 1621 (νC=N); 1605 (νC=C); 1519, 1344 (νNO2); 1296, 1072 (νC-O); 980 (νV=O). UV-Vis spectrum in DMSO : 314 (8430), 345 (3660), 452 (1320). CD spectrum in DMSO : 288 (−2.52), 319 (−3.92), 393 (1.29). 1H NMR : 9.59  (azomethine); 8.32 , 8.25 , 8.07 , 7.85 , 7.69 , 7.59 , 7.41 , 7.20  (aromatic); 5.98 , 3.95  (methine); 4.34 , 4.28  (methylene). 51V NMR : −541.1.S(−)-limonene and (−)-α-pinene, has been studied in the presence of all chiral oxidovanadium(V) complexes using, as the terminant oxidant, aqueous 30% H2O2 or 5.5 M solution of tert-butyl hydroperoxide (TBHP) in decane. After optimization of the reaction conditions, with different amounts of catalysts and oxidants, all reactions were run using 1:100:200 molar ratio of catalyst, substrate, and oxidant, respectively, at 80 °C with in 1,2-dichloroethane (DCE) as the solvent for TBHP and acetonitrile for H2O2. The reaction progress was monitored using GC, and the yields were recorded as GC yield based on the starting substrate. The identity of oxidation products was confirmed using GC-MS.Epoxidation of two alkenes, i.e., styrene and cyclohexene, and monoterpenes, i.e., 3, 4 mM l-glutamine, 1000 mg/L glucose, 1 mM sodium pyruvate, 100 U/mL penicillin and 100 µg/mL streptomycin was employed to culture HT-22 cell line. Cultures were maintained at 37 °C in a humidified atmosphere of 5% CO2. Cells were regularly split and subcultured up to 80–90% confluence (2–3 times per week).The authors want to thank Prof. Tilman Grune from Friedrich Schiller University in Jena (Germany) for a kind gift of the mouse hippocampal neuronal cell line HT-22. Dulbecco’s Modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS), 1.5 g/L NaHCOWorking solutions at 10, 25, 50, and 100 µM concentrations were prepared in serum-free medium DMEM every time before adding to the cell line. The evaluation of the viability and cytoprotective action of investigated compounds on HT-22 hippocampal neuronal cells was tested by using mitochondrial dehydrogenase activity assay, MTT , after 24 h incubation.3 cells in a well of a 96-well plate. The plates were incubated under standard conditions in an incubator for 24 h. After 24 h, HT-22 cells were treated with selected concentrations that were not cytotoxic to cells for 24 h. In experiments studying the cytoprotective effects, investigated compounds were added 1 h before treatment with 30% H2O2 (500 µM) for 24 h. MTT assay was performed by adding a premixed optimized dye solution triazole blue formazan to culture wells. After incubation, the medium was removed, and the formed formazan crystals were dissolved by adding 100 µL DMSO into each well. Next, absorbance was measured at length waves 570 nm and 660 nm  in a microplate reader.HT-22 cells were passaged at 8 × 102O2 (500 µM) and incubated for 24 h. Images were then taken with an Olympus microscope at 1 × 4 magnification.HT-22 cells were seeded in 12-well plates. The plate was stored under standard conditions for 24 h. After 24 h, test compounds were added to HT-22 cells one hour prior to treatment with 30% H1VOL–10VOL, which have been synthesized by monocondensation reaction of salicylaldehyde and its derivatives with 1S,2S-(+)-2-amino-1-(4-nitrophenyl)-1,3-propanediol. All compounds have been characterized using different spectroscopic methods, viz. IR, UV-Vis, circular dichroism, one-  and two-dimensional  NMR spectroscopy, and elemental analysis. In our paper, we described new oxidovanadium(V) complexes with chiral tetradentate Schiff bases, 1VOL–10VOL, have also been employed as catalysts in the epoxidation reactions for testing their catalytic abilities. For this purpose, model olefinic substrates, i.e., styrene, cyclohexene, and its naturally occurring monoterpenes, i.e., S(−)-limonene and (−)-α-pinene, have been studied using the oxygen source two oxidants, i.e., aqueous 30% H2O2 or tert-butyl hydroperoxide. Conversion of all these olefins was distinctly lower when 30% H2O2 was used, but due to the strong oxidizing nature of hydrogen peroxide and the presence of a significant amount of water, which could be responsible for the decomposition of the catalyst and also the hydrolysis of epoxides. On the other hand, tert-butyl hydroperoxide (TBHP) in a non-aqueous environment proved to be a very good oxidizing agent, giving excellent conversions of substrates with epoxide as the main product.Furthermore, these oxidovanadium(V) complexes, cis-dioxidomolybdenum(VI) and oxidovanadium(V) complexes with the same chiral Schiff base ligands towards the hippocampal neuronal cell line HT-22 against the oxidative damage generated exogenously by using hydrogen peroxide. The comparison of the MTT test results between cis-dioxidomolybdenum(VI) and oxidovanadium(V) complexes has shown significantly higher cytotoxic activity in lower concentrations for the latter compounds, which strongly depends on the concentration used. The cytoprotective activity experiments with a 500 µM dose of 30% H2O2 have shown their strong influence on viability at even 100 µM dose in the case of cis-dioxidomolybdenum(VI) compounds. Unfortunately, all investigated oxidovanadium(V) complexes, from 10 to 100 μM concentrations, have not exhibited any cytoprotective effect against the oxidative damage caused by hydrogen peroxide.The biological studies, based on an assessment of mitochondrial activity in MTT tests, have revealed cytoprotective activity of


Flavonoids and chalcones are known for their manifold biological activities, of which many affect the central nervous system. Pyranochalcones were recently shown to have a great neurogenic potential, which is partly due to a specific structural motif-the pyran ring. Accordingly, we questioned if other flavonoid backbones with a pyran ring as structural moiety would also show neurogenic potential. Different semi-synthetic approaches starting with the prenylated chalcone xanthohumol, isolated from hops, led to pyranoflavanoids with different backbones. We identified the chalcone backbone as the most active backbone with pyran ring using a reporter gene assay based on the promoter activity of doublecortin, an early neuronal marker. Pyranochalcones therefore appear to be promising compounds for further development as a treatment strategy for neurodegenerative diseases. Flavonoids are natural products with variable phenolic structures. With over 8000 individual components known, flavonoids can be isolated from a wide range of plants. All flavonoids have in common the basic structure of flavan, however, due to the oxidation and substitution pattern of the C-ring, structural differences may arise . DependiIn the central nervous system, flavonoids were shown to dampen microglial activation, to modulate inflammatory processes , to possRecently, we identified the chalcone xanthohumol C, found in hops, as a potent inducer of neuronal differentiation. Moreover, it is neuroprotective and supports neuroregeneration ,14,15. C1), a prenylated chalcone, is a byproduct of the hop industry, available at a large scale and therefore an ideal educt for further synthesis of prenylated chalcones and flavonoids chromen-4-one): white solid; 1H NMR  δ 1.41 , 2.62 , 2.97 , 3.81 , 5.41 , 5.55 , 6.08 , 6.53 , 6.89 , 7.38 ; 13C NMR  δ 28.22 & 28.53 ((C-7″ & C-8″)), 46.03 (C-2), 56.18 (OCH3), 78.39 (C-6″), 79.78 (C-3), 94.37 (C-6), 103.37 (C-8), 106.46 (C-4a), 116.08 (C-4″), 116.67 (C-3′ & C-5′), 127.14 (C-5″), 128.73 (C-2′ & C-6′), 131.11 (C-1′), 158.45 (C-8a), 159.64 (C-4′), 160.26 (C-7), 163.02 (C-5), 188.26 (C-4).Pyranoflavanone isoxanthohumol C (3) (2 mmol) was dissolved in 10 mL 1,4-dioxane and stirred with 1 mmol DDQ for 3 h at a temperature of 110 °C using conventional oil bath heating. The reaction mixture was poured into 50 mL water and extracted three times with 15 mL EtOAc. The combined organic layers were washed with saturated NaCl-solution and dried over MgSO4. The solvent was removed under reduced pressure and the product was purified via flash chromatography. The yield was 51%.Isoxanthohumol C (5) 2-(4-hydroxyphenyl)-5-methoxy-8,8-dimethyl-4H,8H-pyranochromen-4-one: light yellow solid; IR (ATR) νmax 3429, 1643, 1594, 1515, 1490, 1443, 1401, 1344, 1284, 1200, 1178, 1158, 1132; 1H NMR  δ 10.21 ; 7.86 , 6.89 , 6.58 , 6.44 , 5.78 , 3.80 , 1.45 ; 13C NMR  δ 175.61 (C-4), 160.51 (C-4′), 160.00 (C-5), 159.69 (C-2), 157.04 (C-7), 153.03 (C-8a), 128.04 (C-5′), 127.81 (C-2′ & C-6′), 121.38 (C-1′), 115.92 (C-3′ & C-5′), 114.78 (C-4″), 108.02 (C-4a), 106.03 (C-3), 102.5 (C-8), 96.6; (C-6), 77.90 (C-6″), 56.11 (OCH3), 27.76 (C-7″ & C-8″) HRESIMS m/z 351.1254 ; TLC (n-hexane/EtOAc 1/1) RF 0.05.Pyranoflavone (2) (1 mmol) was dissolved in 10 mL pyridine, 1.2 eq mercury(II)acetate was added and the mixture was stirred for 3 h at 110 °C. The reaction mixture was acidified with HCl (3 M), diluted with 100 mL water and extracted for three times with 15 mL EtOAc. The combined organic layers were washed with saturated NaCl-solution and dried over MgSO4. The solvent was removed under reduced pressure and the product was purified via flash chromatography. The yield was 85% (0.17 mmol). Method E (2) (1 mmol) was dissolved in 10 mL pyridine and 7 mmol silver acetate was added. The reaction mixture was stirred for 48 h at room temperature in the dark. The silver acetate was removed by filtration and the reaction mixture was acidified with HCl (3 M), 30 mL water was added and it was extracted 3 times with 15 mL EtOAc. The combined organic layers were washed with saturated NaCl-solution and dried over MgSO4. The solvent was removed under reduced pressure and the product was purified via flash chromatography. The yield was 39%.Method D : XanthohMethod E : Xanthoh6) 2-(4-hydroxybenzylidene)-4-methoxy-7,7-dimethyl-7H-furochromen-3(2H)-one: yellow solid; IR (ATR) νmax 3311, 2925, 1687, 1658, 1621, 1581, 1516, 1463, 1440, 1416, 1362, 1343, 1263, 1203, 1176, 1158, 1081, 1011, 953, 899, 880, 855, 838, 811, 756, 730, 701, 667, 638, 618, 586, 550, 527, 495, 480, 454, 440; 1H NMR  δ 10.08 , 7.80 , 6.87 , 6.73 , 6.65 , 6.27 , 5.79 , 3.86 , 1.46 ; 13C NMR  δ 178.62 (C-4), 162.26 (C-7), 162.15 (C-8a), 159.67 (C-4′), 159.21 (C-5), 146.20 (C-3), 133.23 (C-2′ & C-6′), 127.96 (C-5″), 123.39 (C-1′), 116.08 (C-3′ & C-5′), 110.76 (C-2). 104.72 (C-4a), 98.97 (C-8), 95.03 (C-6), 78.85 (C-6″), 56.22 (OCH3), 27.94 (C-7″ & C-8″); HRESIMS m/z 351.1251 ; TLC (n-hexane/EtOAc 1/1) Rf 0.47.Pyranoaurone (2) (1 mmol) was dissolved in 3 mL potassium hydroxide (8 M) and 750 µL hydrogen peroxide (30%) was added. The mixture was stirred for 2 h at room temperature, acidified with HCl (3 M) and extracted three times with 10 mL EtOAc. The combined organic layers were washed with saturated NaCl-solution and dried over MgSO4. The solvent was removed under reduced pressure and the product was purified via flash chromatography und recrystallized from acetonitrile/water 1:1. The yield was 20%.Xanthohumol C (7) 3-hydroxy-2-(4-hydroxyphenyl)-5-methoxy-8,8-dimethyl-4H,8H-pyranochromen-4-one: yellow solid; IR (ATR) νmax 3433, 3214, 2923, 1610, 1575, 1562, 1513, 1448, 1425, 1361, 1334, 1284, 1248, 1220, 1201, 1175, 1153, 1124, 1023, 839, 825, 586; 1H NMR  δ 9.99 , 8.78 , 8.01 , 6.42 , 5.76 , 3.84 , 1.44 ; 13C NMR  δ 171.05 (C-4), 159.95 (C-5), 128.72 (C-2′ & C-6′), 127.74 (C-5″), 121.95 (C-1′), 115.53 (C-3′ & C-5′), 114.66 (C-4″), 106.07 (C-4′), 101.68 (C-8), 95.95 (C-6), 77.98 (C-6″), 56.25 (OCH3), 27.77 (C-7″ & C-8″); HRESIMS m/z 367.1204 ; TLC (n-hexane/EtOAc 1/1) Rf 0.28.Pyranoflavonol (v/v) B27 , 2 mM L-glutamine, 100 U/mL Penicillin, 100 μg/mL Streptomycin, 20 ng/mL Epidermal Growth Factor-(rhEGF), 20 ng/mL Fibroblast Growth Factor (rhFGF) , 2 µg/mL Heparin and maintained in a humidified atmosphere at 37 °C and 5% CO2.The assay was performed as previously described in Oberbauer et al. . In brieRenilla luciferase under the CMV promoter to normalize the DCX-driven gene expression. Transfected cells were resuspended in NB, supplemented with 2% (v/v) B27, 2 mM L-glutamine, 100 U/mL Penicillin, 100 μg/mL Streptomycin and 1% fetal calf serum (FCS), and seeded into 100 µg/mL poly-ornithine/5 µg/mL laminin coated white 96-well plates . One day after cell seeding, the medium was replaced by fresh NB-B27 with 1% FCS containing 10 µM of the flavonoids to be tested. Each substance was applied for three days in technical tetraplicates. The bioluminescent measurement was performed in a TriStar Multimode Microplate Reader LB 941  according to the manufacturer’s instructions.Luciferase assays were performed using a non-commercial dual luciferase enzyme assay system . In thisFor one-way ANOVA with Bonferroni test, OriginPro 2021b (Originlab Corporation) was used.In this study, different pyranoflavonoids were produced via semi-synthetic approaches and the neurogenic potential of the different flavonoid backbones was investigated. The semi-synthetic yields were improved by microwave irradiation compared to the conventional heating method. Furthermore, the possibility of using silver acetate for aurone synthesis instead of toxic reagents such as thallium or mercury acetate is noteworthy. The various semi-synthetic isomerization reactions are suitable for further structure-activity studies of natural products, as mostly simple one-pot, one-step reactions were used, providing an easily accessible source of different flavonoid backbones. Furthermore, we have shown that the chalcone backbone, in addition to the pyran ring, is beneficial for neurogenic activity according to the DCX promoter reporter assay. Therefore, pyranochalcones appear to be promising compounds for further development as a strategy for the treatment of neurodegenerative diseases.


JBP) in Bahía Solano, Chocó, focused on studying the diversity of the mygalomorph spider fauna, allowed us to discover four new species included in the families Halonoproctidae and Theraphosidae. The trapdoor species Ummidiasolanasp. nov., and the theraphosids species Euthycaeluscunampiasp. nov. (Schismatothelinae), Melloinapacificasp. nov. (Glabropelmatinae), and Neischnocolusmecanasp. nov. (Theraphosinae) are illustrated, diagnosed, and described in detail. Photographs of somatic features and copulatory organs and a distribution map are provided. Morphological, taxonomical, and biogeographical aspects are discussed for each species. All these taxonomic novelties represent the first records of these genera for the region, expanding the range of geographic distribution of each of them. This work constitutes the first effort focused on characterizing the community of Mygalomorphae species in the Chocó Biogeographic Region.The Colombian Pacific coast is an amazing natural region, immersed in one of the most unknown biodiversity hotspots in the world. An expedition carried out in the north of this area, at the Jardín Botánico del Pacífico ( Mygalomorphae spiders, known in a broad sense as tarantulas in America, constitute approximately 6% of the total number of spider species described , focused on Mygalomorphae spiders. The JBP, located in Bahía Solano, is a tourist area and natural reserve that plays a key role in the conservation of the tropical rainforest and mangroves of the Colombian Chocó Biogeographic Region , Neischnocolusmecana sp. nov. (Theraphosinae), and Melloinapacifica sp. nov (Glabropelmatinae). All these new taxonomic records represent first reports of these genera for the region, which extends the range of geographical distribution of each of them. The results of this work constitute a contribution to the knowledge of the biological diversity of one of the areas with the greatest specific richness of species and endemism in Colombia.In this first approach to the knowledge of the Araneae) of the Instituto de Ciencias Naturales (ICN), Universidad Nacional de Colombia, Bogotá, Colombia, preserved in 75% ethanol. The specimens were collected during a biological expedition carried out in the Jardín Botánico del Pacífico (JBP), located in Bahía Solano, Chocó, Colombia ; and deposited in the Arachnological Collection  by immersion in a test tube and subjecting them to increased heat for short time intervals. Setae of the male tibia I and palpal tibia were removed in order to illustrate the tibial apophysis and nodules, respectively. Specimens and the structures removed were examined under a LEICA M205C stereo microscope. Photographs were taken with a stereo microscope ZEISS Stereo Discovery V12, then stacked with Helicon Focus 8.2.0 Mac OS (Helicon Soft Ltd. 2019) and processed with Adobe Photoshop CC 2022 (Adobe Inc. 2022).All measurements are given in millimeters (mm). The total length given does not include the chelicerae or spinnerets. Eye sizes were measured as the maximum diameter in either a dorsal or frontal view and were taken with a digital micrometer. Body measurements were taken with a digital micrometer or a vernier caliper. The length and width of carapace, eye tubercle, labium and sternum are the maximum values obtained. Leg and palp measurements were taken in dorsal view along the central axis of the right-side limbs and were taken with a vernier caliper.Ummidia and Ummidiasolana sp. nov. is based on Melloinapacifica is based on Ummidia and Melloina are considered as pseudoscopula, according to Theraphosinae and Schismatothelinae, respectively. Urticating setae terminology follows JBP are recorded in the original description: 6.38, -77.40; specific geographic location data are recorded in the collection label for each of them. The distribution map was produced using QGIS 3.26.1 – Buenos Aires.The general descriptive format follows Abbreviations used in the text and figures are as follows:A apical keel of palpal bulb;ALE anterior lateral eyes;AME anterior median eyes;ap apical;d dorsal side;ITC inferior tarsal claw;p prolateral side;PB prolateral branch of tibial apophysis;PI prolateral inferior keel of palpal bulb;PLE posterior lateral eyes;PME posterior median eyes;PMS posterior median spinnerets;PLS posterior lateral spinnerets;PS prolateral superior keel of palpal bulb;R retrolateral keel of palpal bulb;RB retrolateral branch of tibial apophysis;SLS spine like setae;SR seminal receptacles;STC superior tarsal claw;v ventral side.Ummidia Thorell, 1875Genus Taxon classificationAnimaliaAraneaeHalonoproctidaeF2736434-F432-5DE7-BC46-75498A479E49https://zoobank.org/C21B3C70-626A-4B19-830E-25B8E74EC200Holotype ♂: Colombia, Chocó, Bahía Solano, Jardín Botánico del Pacífico, 6.38, -77.40, elevation 60 m a.s.l., 10–25 February 2022, M. Echeverri, S. Gómez Torres and C. Perafán leg. (ICN 12356). Paratype ♀: same data as holotype, except elevation 132 m a.s.l. (ICN 12357).solana is a noun in feminine refers to the municipality of Bahía Solano, one of the most beautiful places in the Colombian Pacific coast, recognized for having large and desolate beaches and landscapes of abundant vegetation. It is immersed in one of the world’s biodiversity hotspots. It is also said that the word “solano” means “wind from where the sun rises”.The specific epithet Ummidiasolana sp. nov. can be differentiated from all geographically proximate species (see U.quijichacaca and U.tibacuy unknown). Female: oval carapace, longer than wide, with strongly procurved fovea, wide and deep ; basal segment of chelicerae with numerous lateral teeth (9–10) (4–6 in U.quijichacaca and U.tibacuy); maxillae with two sets  of few cuspules of similar number (ca. 12) ; palp trochanter with distinct group of elongated cuspules ; tarsus IV with comb of long spinules on the retrolateral face ; and seminal receptacles straight oriented inwardly, mushroom-shaped distally subspherical, with wide rounded bulbs . Additionally, female and male  with black carapace and legs, grayish abdomen, and male with reddish brown tarsi  Fig.   ; basal segments with fine, small, and macro-spigots, distal segment with numerous fine spigots and few macro-spigots. PMS with one segment, length 0.65, with numerous small spigots.  Figs , 4–7. ToAME 0.36 (circular), ALE 0.42 , PME 0.19 (circular), PLE 0.29 , AME-AME 0.09, AME-ALE 0.09, ALE-ALE 0.94, PME-PME 0.49, PME-PLE 0.04, PLE-PLE 0.97, AME-PME 0.05, ALE-PLE 0.10. Thoracic fovea  ; basal segments with fine, small, and macro-spigots, distal segment with numerous fine spigots and few macro-spigots. PMS with one segment, length 0.78, with numerous small spigots.  Figs , 8–10. TAME 0.3 (circular), ALE 0.52 , PME 0.5 , PLE 0.42 , AME-AME 0.13, AME-ALE 0.42, ALE-ALE 1.15, PME-PME 0.45, PME-PLE 0.11, PLE-PLE 1.15, AME-PME 0.17, ALE-PLE 0.27. Thoracic fovea  along 60% of the segment; patella with a low promedial lobe and four short and wide spines; tibia with a prolateral and retrolateral wide row of curved spines along full length of segment; metatarsus-tarsus with a prolateral and retrolateral wide row of curved spines along full length of segment. Femora I and II: with central rows of short fine SLS along full length of segment. Patellae I and II: with a low retromedial lobe, with three dorsal central rows of fine and short SLS along full length of segment, with some ventral long SLS. Tibiae I and II: with several row of dorsal SLS; tibia I with a wide prolateral and retrolateral row of curved spines along full length of segment; tibia II with a ventral row of lightly curved spines along full length of segment, with a prolateral row of short, curved spines along full length of segment. Metatarsi I and II: with dorsal SLS along full length of segment, and a prolateral and retrolateral row of curved spines along full length of segment. Tarsi I and II: with dorsal SLS and a prolateral and retrolateral row of short, curved spines.Legs I Fig.  and II, SLS. Patella: dorsal with a central row of SLS along full length of segment, with ca. 13 prodorsal short and strong spines in distal edge. Tibia . Paratypes, same data as holotype except elevation, 99–145 m a.s.l.: ♀ (ICN 12359), ♂ (ICN 12360), ♂ (ICN 12361), ♀ (ICN 12362), ♂ (ICN 12363).pacifica is a noun in feminine refers to the Colombian Pacific region, where the species is distributed.The specific epithet Melloinapacifica sp. nov. can be distinguished from other Melloina species by the relatively longer embolus , > 3 × in M.santuario Bertani, 2013), by the labium with numerous cuspules (112 vs. 60–80 in other known species), and by the tarsi II, III, IV ventrally cracked at midpoint . Additionally, differs from M.santuario by the number of spines on tarsi I and II . Female can be distinguished from other Melloina species by the straight, long, and wide spermathecae, without glandular area in the basal third , and labium with 98 cuspules .Male of lus Fig. , ca. 2 ×ird Fig. , numerouKnown only from the type locality Figs , 2.Male (holotype) ; PMS with one segment, length 0.45.  Figs –17. Tota45. Fig. .AME 0.39 (circular), ALE 0.46 , PME 0.30 , PLE 0.35 , AME-AME 0.09, AME-ALE 0.19, PME-PME 0.72, PME-PLE 0.09, PLE-PLE 1.31, ALE-PLE 0.11, AME-PME 0.05. Thoracic fovea , r 0-0-5; II d 5-5-7, p 0-0-1, v 0-0-6 (5 ap), r 1-1-1; III d 5-5-4, p 3-2-2, v 1-3-10 (8 ap), r 1-2-3; IV d 5-2-4, p 0-0-1 ap, v 0-2-10 (8 ap), r 0-1-1. Patellae: palp d 9-7-8, p 0-1-4 ap, v 1-1-4 ap, r 2-3-2; I d 1-1-3, p 1-1-2, v 0-0-2 ap, r 1-1-2 ap; II d 3-2-5, p 1-2-5 (4 ap), v: 0-1-3 ap, r: 1-1-2; III d: 5-4-5, p: 2-2-2, v: 0-2-2 ap, r: 1-1-3 ap; IV d: 3-1-2, p: 0-1-2 ap, v 0-2-3 (2 ap), r 0-0-1 ap. Tibiae: palp d 2-5-5, p 4-2-4 (2 ap), v 5-6-7 (3 ap), r 2-3-8 (2 ap); I d 1-1-3, p 1-1-2, v 0-0-2 ap, r 1-1-2 ap; II d 2-1-1, p 3-2-3 (2 ap), v 4-4-4 (2 ap), r 2-2-0; III d 3-2-3, p 3-2-3 (1 ap), v 2-2-2 ap, r 3-3-3 ap; IV d 7-5-7, p 5-4-4 (1 ap), v 3-2-3 (2 ap), r 2-2-2 (1 ap). Metatarsi: I d 0, p 2-2-1 ap, v 6-4-6 (2 ap), r 1-1-1; II d 2-2-2, p 4-4-4 (1 ap), v 4-5-6 (3 ap), r 3-2-2 (1 ap); III d 4-4-3, p 2-2-2, v 4-4-5 (2 ap), r 2-3-3 (1 ap); IV d: 3-1-4, p: 2-2-2, v: 3-3-4 (2 ap), r: 3-2-2 (1 ap). Tarsi: cymbium p lobe 4, r lobe 10 (8 ap); I d 0, p 3-1-4 (1 ap), v 0, r 3-3-4 (1 ap); II d 1-1-1, p 3-3-4 (1 ap), v 0, r: 3-3-4 (1 ap); III d 3-2-3, p 3-3-4 (1 ap), v 0, r 3-3-4 (1 ap); IV d 3-4-3, p 2-3-3 (1 ap), v 2-1-2, r 2-3-3 (1 ap).Spination . Femora: palp d 5-8-11, p 0-0-3, v 0-2-0, r 0-2-2; I d 6-4-10, p 0-0-3, v 3-4-10  ; PMS with one segment, length 0.69.  Figs , 19. Tot69. Fig. .AME 0.32 (circular), ALE 0.48 , PME 0.23 (circular), PLE 0.37 , AME-AME 0.09, AME-ALE 0.14, PME-PME 0.8, PME-PLE 0.09, PLE-PLE 1.57, ALE-PLE 0.13, AME-PME 0.06. Thoracic fovea , r 0. Tibiae: palp d 5-5-3, p 0-1-2, v 6-5-3 ap, r 2-2-2; I d 4-6-4, p 2-1-2, v 3-4-2 ap, r 1-1-1; II d 2-1-1, p 1-1-2, v 4-5-4 (2 ap), r 1-1-1; III d 4-5-5, p 2-2-2, v 3-4-4 (3 ap), r 0-1-1; IV d 2-2-2, p 3-2-1 ap, v 4-5-5 (1 ap), r 1-1-2 (1 ap). Metatarsi: I d 0, p 1-1-0, v 5-5-6 (2 ap), r 0; II d 0, p 1-1-3, v 5-3-5 (2 ap), r 1-2-2 (1 ap); III d 2-3-4, p 2-2-2 (1 ap), v 4-3-3 (1 ap), r 2-2-2; IV d 4-4-4, p 2-2-1, v 4-3-4 ap, r 2-2-2. Tarsi: palp d 0, p 3-3-3, v 1-3-0, r 2-3-2; I d 0, p 3-3-4, v 0-0-2 ap, r 2-3-2; II d 0, p 3-3-3 (1 ap), v 0, r 3-3-3; III d 0, p 3-3-3, v 0, r 3-4-3; IV d 0, p 3-3-4 (1 ap), v 0-3-0, r 4-3-4 (1 ap).Spination . Femora: palp d 2-2-4, p 0-0-1, v 12-9-8, r 0; I d 3-2-3, p 0, v 3-3-5, r 0; II d 1-0-1, p 0, v 4-3-3, r 0; III d 1-2-1, p 1-1-1, v 1-2-3, r 0; IV d 1-1-1, p 0, v 1-2-6, r 0. Patellae: palp d 3-2-3, p 1-1-3 Spermatheca Fig. : two semColoration. In alcohol: as described in the male.Melloinapacifica sp. nov. is the first species of the genus described for Colombia, although it is known that Melloina is distributed in different ecosystems, including cave environments . Additionally, its taxonomic position is currently being widely debated .cunampia is a patronym in honor of the family name of Don José and Don Antonio, members of the Emberá indigenous community, from Mecana, Chocó. Mr. José and Mr. Antonio abandoned their hunting traditions for their community to become touristic and academic guides for the JBP. We want to pay tribute to their community and to the JBP with this recognition.The specific epithet Euthycaeluscunampia sp. nov. can be distinguished from all other Euthycaelus species by the following combination of morphological features: the shape of the palpal bulb , and additionally by a higher number of maxillary cuspules (ca. 200 vs. 150) and labial cuspules (ca. 300 vs. 200). Additionally, males and females  with carapace and legs black covered by light brown setae, and tibiae, metatarsi and tarsi covered with very light setae ; PMS with one segment, length 1.0. , ALE 0.50 , PME 0.39 (subcircular), PLE 0.45 , AME–AME 0.11, AME–ALE 0.13, ALE–ALE 1.22, PME–PME 0.98, PME–PLE 0.02, PLE–PLE 1.47, AME–PME 0.04, ALE–PLE 0.13. Thoracic fovea , II distal 3/4 (75%), III more than distal half (60%), IV distal half (50%) sparsely scopulated proximally. Trichobothria: tarsi with two rows of clavate trichobothria, each with ca. 25, interspersed with filiform trichobothria of different sizes. Tarsus IV slightly cracked at midpoint. Femur III: slightly incrassate. Plumose setae on retrolateral face of femur IV: absent. Stridulatory bristles: absent. Urticating setae: absent.Legs pattern: IV>I>II>III. Lengths of legs and palpal segments on Table ap), p 0-0-1; III d 1-0-2, v 1-2-2 (ap); IV d 2-1-2, v 2-2-3 (ap). Metatarsi: I v 1-0-3 (ap); II v 1-0-3 (ap); III d 1-2-2, v 1-2-3 (ap); IV d 1-2-2, v 1-2-4 (3 ap).Spination . Cymbium and tarsi without spines. Femora: palp d 0-0-1p; I d 0-0-1p; II d 0-0-1p; III d 0-0-2p-r; IV d 0-0-1r. Patella: I–II, IV and palp 0; III r 0-0-1d. Tibiae: palp p 0-0-1d, r 0-0-7; I v 0-0-1, p 0-0-1; II v 0-0-2 . Paratype ♀: same data as holotype (ICN 12366).mecana is a noun in apposition related to one of the townships of the municipality of Bahía Solano, where the JBP is located. The name of this small town is due to the fact that it is located on the Mecana riverside, with crystalline waters and abundant biodiversity. The JBP promotes the conservation, research, and recovery of the native biodiversity of this region. We would like to pay tribute to its community and the JBP with this recognition.The specific epithet Neischnocolusmecana sp. nov. can be distinguished from the other Neischnocolus species by the following combination of morphological characters: shape of the palpal bulb piriform, with the tip of the embolus continuing the palpal organ axis (not perpendicular), well-developed prolateral (PS and PI) and apical (A) keels with non-serrated edge, PI discontinuous, absence of retrolateral keel (R), and without granulation or microspikes on embolus or tegulum ; PMS with one segment, length 1.14. , ALE 0.68 , PME 0.40 , PLE 0.55 , AME-AME 0.30, AME-ALE 0.18, PME-PME 1.11, PME-PLE 0.19, PLE-PLE 1.80, ALE-PLE 0.24, AME-PME 0.16. Thoracic fovea: transverse, width 3.25; slightly procurved, deep, 8.75 from the anterior edge of carapace. Chelicerae basal segment: length 4.2, width 3.1; with 11 well-developed teeth on each furrow promargin, and a group of ca. 15 small teeth near last three basal promargin teeth. Maxillae , p 1-1-1, r 0-1-1; IV d 0-0-1, v 2-3-4 (3 ap), p 1-1-1, r 0-1-1.Spination: All femora, patellae, and tarsi 0. Legs I–II and palp 0. Tibiae: I–II 0; III d 0-0-1, v 0-1-2 PS and PI) and apical keel (A) present, PS discontinuous; without granulation on embolus or tegulum; tegular apophysis developed. Palpal tibia with two distinct subconical distal processes on retrolateral surface ; PMS with one segment, length 2.05. , ALE 0.70 , PME 0.41 , PLE 0.50 , AME-AME 0.34, AME-ALE 0.30, PME-PME 1.27, PME-PLE 0.26, PLE-PLE 2.00, ALE-PLE 0.36, AME-PME 0.21. Thoracic fovea: transverse, width 3.50; slightly procurved, deep, 9.11 from the anterior edge of carapace. Chelicerae basal segment: length 5.1, width 3.6; with 11 well-developed teeth on each furrow promargin, and a group of ca. 12 small teeth near last three basal promargin teeth. Maxillae .Urticating setae: types I urticating setae present, subtype Ic  and I moc  / subtypc , p 1-2-0, r 0-1-1.Spination: All femora, patellae, and tarsi 0. Tibiae: palp 0-0-2 Spermathecae Fig. : consistColoration. Living spiders: carapace, abdomen, and legs black, legs with light-colored stripes at the joints. Ventral abdomen with patterns of dark spots (Fig. ots Fig.  and coxaots Fig. . In alcoNeischnocolusmecana sp. nov. it is the fourth species of the genus described for Colombia and it is the first record of Neischnocolus for the Chocó biogeographic region, as well as the first record for the Colombian Pacific. With this description, the known geographic range of the genus is extended. Currently, Neischnocolus is distributed in Colombia in the Andean, Amazonian, and Pacific regions. It is known that Neischnocolus is widely distributed in the Colombian territory, with a very extensive geographical and altitudinal range, and that most of its diversity has not yet been described (escribed .


Furthermore, HMC dyes with broad absorption in the near-infrared region are synthesized by combining a trifluoromethylated anionic HMC dye with a cationic HMC dye as a counterion.A novel anionic heptamethine cyanine (HMC) dye with two trifluoromethyl groups that selectively absorb near-infrared light is synthesized. When contrasted with previously studied anionic HMC dyes with substituents such as methyl, phenyl, and pentafluorophenyl groups, the trifluoromethylated dye displays a red-shifted maximum absorption wavelength (for instance, 948 nm in CH However, comparatively lower photostability hinders their application +, 174.1254; 1H NMR (DMSO-6d) δ 1.54 (C3H)), 2.79–2.80 , 3.99 , 7.58–7.65 , 7.81–7.88 , 7.89–7.95 ; 13C NMR (DMSO-6d) δ 14.5 (s), 21.7 (s), 34.9 (s), 53.9 (s), 115.1 (s), 123.3 (s), 128.8 (s), 129.2 (s), 141.6 (s), 142.1 (s), 195.9 (s)  1609 (C=N) cm (s) see .E)-2-chloro-3-(hydroxymethylene)cyclohex-1-ene-1-carbaldehyde (3)  was added to a N,N-dimethylformamide solution (3 mL) of 1,2,3,3-tetramethyl-3H-indol-1-ium iodide (6) , and the mixture was stirred at 120 °C for 5 h. The reaction mixture was poured into ice water (100 mL), stirred for 30 min, and thereafter refrigerated for 30 min. The resulting precipitate was collected using suction filtration. The residue was purified using silica gel chromatography (dichloromethane/methanol = 25/1) to yield 2-((E)-2-((E)-2-chloro-3-(2-((E)-1,3,3-trimethylindolin-2-ylidene)ethylidene)cyclohex-1-en-1-yl)vinyl)-1,3,3-trimethyl-3H-indol-1-ium iodide (7) . ; IR (KBr) 1550 (C=N) cm−1; HRMS (ESI) found: m/z 483.2561 Calc. for C32H36N2Cl: [M − I−]+, 483.2567; 1H NMR (CDCl3) δ 1.77 2 × 2), 1.96–1.99 , 2.75–2.81 , 3.76 , 6.25 , 7.17–7.24 , 7.34–7.39 , 8.34 ; 13C NMR (CDCl3) δ 20.8 (s), 27.0 (s), 28.2 (s), 32.8 (s), 49.3 (s), 102.0 (s), 110.9 (s), 122.2 (s), 125.4 (s), 128.1 (s), 129.0 (s), 141.1 (s), 143.0 (s), 144.4 (s), 150.6 (s), 173.0 (s)  see .7  and 5a  in acetone (2 mL) was stirred at 25 ℃ overnight under an argon atmosphere. After the solvent was removed under low pressure, the residue was purified via column chromatography on silica gel using a CH2Cl2–methanol (200:1 (v/v)) solvent to obtain 2-((E)-2-((E)-2-chloro-3-(2-((E)-1,3,3-trimethylindolin-2-ylidene)ethylidene)cyclohex-1-en-1-yl)vinyl)-1,3,3-trimethyl-3H-indol-1-ium ((Z)-4-((E)-2-(2-chloro-3-((E)-2-(4-cyano-5-(dicyanomethylene)-2-methyl-2-(trifluoromethyl)-2,5-dihydrofuran-3-yl)vinyl)cyclohex-2-en-1-ylidene)ethylidene)-3-cyano-5-methyl-5-(trifluoromethyl)-4,5-dihydrofuran-2-yl)dicyanomethanide (8a) with a yield of 92% (0.011 g).A mixture of Tdt 201.3 °C; fR 0.55 (CH2Cl2/methanol = 10/1); IR (KBr) 1550 (C=N), 2214 (C≡N) cm−1; HRMS (ESI) found: m/z 483.2566. Calc. for C32H36ClN2: [M]+, 483.2567, m/z 641.0931. Calc. for C30H16ClN6O2F6: [M]−, 641.0927; 1H NMR (Acetone-d6) δ 1.78 , 1.83–1.88 , 1.93 , 1.94–1.99 , 2.55–2.71 , 2.77 , 3.80 , 6.24 , 6.41 , 7.32 , 7.41–7.49 , 7.62 , 8.44 , 8.55 ; 13C NMR (Acetone-d6) δ 19.6 (s), 21.6 (s), 26.9 (s), 27.0 (s), 28.1 (s), 32.0 (s), 49.6 (s), 50.1 (s), 85.8 (s), 92.8 , 102.5 (s), 109.3 (s), 112.0 (s), 113.9 (s), 114.5 (s), 114.6 (s), 123.2 (s), 123.9 , 126.2 (s), 127.4 (s), 130.0 (s), 131.3 (s), 141.6 (s), 142.1 (s), 144.1 (s), 144.7 (s), 149.4 (s), 150.1 (s), 156.7 (s), 174.3 (s), 177.1 (s); 19F NMR (Acetone-d6) δ −80.25   see .−3 M) containing Bu4NClO4 (0.1 M). The Eox values were measured using three small electrodes. A silver quasi-reference electrode, a platinum wire, and a carbon electrode were used as the reference, counter, and working electrodes, respectively. All the electrode potentials were calibrated concerning the Fc/ferrocenium redox couple. Electrochemical measurements were performed at a scan rate of 200 mV s−1. The Eox value of Fc vs. SCE was 0.380 V [Eox values vs. SCE were determined using the observed Eox (V vs. Ag) values of the dyes in MeCN solutions as follows:Electrochemical measurements of the dyes were performed in MeCN solutions (1.0 × 10 0.380 V . The EoxEox (V vs. SCE) values as follows [The energy of the HOMO (eV) was obtained using the  follows :HOMO (eVE0-0) and energy of the LUMO (eV) were calculated using the λonsetabs value as follows:The band gap (2Cl2 solutions of the dyes were maintained in an incubator at 25 °C and irradiated with white LED light (8.5 W).CH5a with trifluoromethyl groups in the dye skeleton and compared its properties with those of our previously synthesized anionic HMC dyes. The new anionic HMC dye 5a showed a more red-shifted absorption wavelength and improved photostability than our previously synthesized anionic HMC dyes. These properties are attributed to the electron-withdrawing characteristics of the trifluoromethyl groups of 5a, which are more potent than the substituents of the previously synthesized dyes. We synthesized a novel anionic HMC dye 8a, composed of an anionic HMC skeleton bearing trifluoromethyl groups and a cationic HMC skeleton. Subsequent investigations into its absorption properties and photostability were conducted. Our findings revealed that the unique properties of both the anionic and cationic HMC skeletons were distinctly represented in this compound. In the cyanine–cyanine mixed dye 8a, the photostability of the cationic HMC skeleton was enhanced by the photodegradation of the anionic HMC skeleton. The enhanced properties of compounds 5a and 8a indicated their potential suitability for photovoltaic devices, thus presenting a significant advantage. We proceeded to synthesize a cyanine–cyanine mixed dye, 3 groups.We are currently investigating organic solar cells that utilize only near-infrared light using HMC dyes with CF