Potential Biological Activity Research Articles

Open and reusable annotated mass spectrometry dataset of a chemodiverse collection of 1,600 plant extracts.

As privileged structures, natural products often display potent biological activities. However, the discovery of novel bioactive scaffolds is often hampered by the chemical complexity of the biological matrices they are found in. Large natural extract collections are thus extremely valuable for their chemical novelty potential but also complicated to exploit in the frame of drug-discovery projects. In the end, it is the pure chemical substances that are desired for structural determination purposes and bioactivity evaluation. Researchers interested in the exploration of large and chemodiverse extract collections should thus establish strategies aiming to efficiently tackle such chemical complexity and access these structures. Establishing carefully crafted digital layers documenting the spectral and chemical complexity as well as bioactivity results of natural extracts collections can help prioritize time-consuming but mandatory isolation efforts. In this note, we report the results of our initial exploration of a collection of 1,600 plant extracts in the frame of a drug-discovery effort. After describing the taxonomic coverage of this collection, we present the results of its liquid chromatography high-resolution mass spectrometric profiling and the exploitation of these profiles using computational solutions. The resulting annotated mass spectral dataset and associated chemical and taxonomic metadata are made available to the community, and data reuse cases are proposed. We are currently continuing our exploration of this plant extract collection for drug-discovery purposes (notably looking for novel antitrypanosomatids, anti-infective and prometabolic compounds) and ecometabolomics insights. We believe that such a dataset can be exploited and reused by researchers interested in computational natural products exploration.

STUDY OF THE EFFECT OF NEW 6-R-3,5-DINITRO-1,2,3,4-TETRAHYDROPYRIDINES ON THE ACCUMULATION OF PHOTOSYNTHETIC PIGMENTS AND ASCORBIC ACID IN PLANT TISSUES

Background. The problem of finding new effective plant protection products remains an urgent task of modern agrochemistry. It is known that tetrahydropyridine derivatives exhibit a broad spectrum of biological activity, including fungicidal activity. However, compounds with potential biological activity can also be highly toxic to humans, cultivated crops, and the environment as a whole, so it is necessary to study their environmental safety. The content of photosynthetic pigments and ascorbic acid are one of the most important biochemical indicators of plant response to environmental stress factors, therefore, determining their amount helps to assessthe physiological state of plants and analyze the toxicity of the synthesized compounds.
 Purpose. To study the effect of new 6-R-3,5-dinitro-1,2,3,4-tetrahydropyridines on the content of photosynthetic pigments and ascorbic acid in plant tissues in order to assess the safety of their use as potential plant protection products.
 Materials and methods. The test compounds were obtained by the previously developed method for the selective reduction of 2-R-3,5-dinitropyridines. Winter wheat seeds of the Moskovskaya 39 variety were germinated in Petri dishes using solutions of the synthesized substances. Seeds germinated in water were used as controls. On the ninth day, the content of various groups of photosynthetic pigments (chlorophylls a, b, carotenoids), as well as ascorbic acid in plant tissues was determined. All experiments were carried out in triplicate. In addition, the analytes were tested for toxicity using the GUSAR Environmental Toxicity software.
 Results. The effect of new derivatives of tetrahydropyridine on the accumulation of photosynthetic pigments and ascorbic acid in plant tissues of winter wheat was studied. It was shown that the studied compounds do not have a significant toxic effect on plants.
 Conclusion. As a result of studying the effect of new 6-R-3,5-dinitro-1,2,3,4-tetrahydropyridines on the content of photosynthetic pigments and ascorbic acid, as well as in silico computer prediction in GUSAR Environmental Toxicity, it was found that, in general, the studied compounds are relatively safe and, therefore, promising for further testing as biologically active plant protection products.

Potential Use of Cardunculus Biomass on Pleurotus eryngii Production: Heteroglycans Content and Nutritional Properties (Preliminary Results)

The new perspective of using waste biomass to cultivate mushrooms as a source of protein for human nutrition, in line with the circular economy principles, is receiving increasing attention in the scientific community and represents great wealth in terms of environmental sustainability. Pleurotus eryngii is a mushroom also known as cardunculus mushroom due to its ability to grow on this plant. This study explores the potential intrinsic properties of cardunculus (for example, the presence of inulin in the roots) as raw material for the growth of cardunculus mushrooms, and the influence on heteroglycan content and nutrition parameters of the fruiting bodies. Both mycelium and fruiting bodies were used to determine the heteroglycan content in the presence of inulin or cardunculus roots rich in inulin. To produce heteroglycans from P. eryngii in greater quantities and shorter times without having to wait for the formation of the fruiting bodies, the mycelium could be used. The results showed that the presence of cardunculus biomass positively influences the heteroglycan content of P. eryngii. In terms of nutritional parameters, higher contents of polyphenols, flavonoids, anthocyanins, and antioxidant activity were detected in P. eryngii grown on the cardunculus stem and root substrate. In conclusion, recycling cardunculus biomass to generate growth blocks for edible mushrooms is a winning choice due to the opportunity to use this biomass waste, which is gaining more and more attention due to the increase in cultivated areas and the use of fruiting bodies of P. eryngii as a functional food and source of molecules with potential biological activities.

Crude Extracts and Secondary Metabolites of Epichloë bromicola against Phytophthora infestans.

Potato late blight caused by Phytophthora infestans is still one of the main factors limiting potato production. Epichloë spp. can provide host plants with various resistances, which makes them show great potential in the biological control of diseases. In this study, we explored the potential biological activity of crude extracts of 20 strains of Epichloë bromicola to control P. infestans. The crude extracts of strains 1 and 8 showed significant antifungal activity with an inhibition rate of 88 % and 81 %, respectively, and showed different effects on the mycelium morphology of P. infestans observed by scanning electron microscopy. Moreover, the two crude extracts demonstrated an interesting therapeutic and protective effect on potato late blight, and none of the extracts had an adverse effect against zebrafish embryos. A total of 13 metabolites were isolated from the crude extract of strain 8, and these tested compounds showed a weak antifungal effect and the inhibition rate was less than 80 %. These findings suggested that strains 1 and 8 have potential for biocontrol of late potato blight.

Molecular Docking of Pharmaceutical Cocrystal As a Ligand: Do We Need to Rethink?

Molecular docking is a resourceful virtual screening tool for predicting the possible therapeutic activities of molecules and lowering the attrition rate at the early stage of drug design and development. Recently reports of molecular docking of cocrystals have been published to demonstrate their potential biological activities toward specific target proteins. Since regulatory bodies, e.g., U.S. FDA and EMA, do not identify cocrystal as a new molecular entity, it is not relevant to conduct such docking studies, especially in the case of orally administered cocrystals, by posing them as ligands instead of the parent API. This article showcases the noteworthy considerations regarding molecular docking studies of cocrystals as ligands.

Predicting Endocrine Disruption Using Conformal Prediction - APrioritization Strategy to Identify Hazardous Chemicals with Confidence.

Receptor-mediated molecular initiating events (MIEs) and their relevance in endocrine activity (EA) have been highlighted in literature. More than 15 receptors have been associated with neurodevelopmental adversity and metabolic disruption. MIEs describe chemical interactions with defined biological outcomes, a relationship that could be described with quantitative structure-activity relationship (QSAR) models. QSAR uncertainty can be assessed using the conformal prediction (CP) framework, which provides similarity (i.e., nonconformity) scores relative to the defined classes per prediction. CP calibration can indirectly mitigate data imbalance during model development, and the nonconformity scores serve as intrinsic measures of chemical applicability domain assessment during screening. The focus of this work was to propose an in silico predictive strategy for EA. First, 23 QSAR models for MIEs associated with EA were developed using high-throughput data for 14 receptors. To handle the data imbalance, five protocols were compared, and CP provided the most balanced class definition. Second, the developed QSAR models were applied to a large data set (∼55,000 chemicals), comprising chemicals representative of potential risk for human exposure. Using CP, it was possible to assess the uncertainty of the screening results and identify model strengths and out of domain chemicals. Last, two clustering methods, t-distributed stochastic neighbor embedding and Tanimoto similarity, were used to identify compounds with potential EA using known endocrine disruptors as reference. The cluster overlap between methods produced 23 chemicals with suspected or demonstrated EA potential. The presented models could be utilized for first-tier screening and identification of compounds with potential biological activity across the studied MIEs.

One‐Pot Synthesis of Thiocarbamide Esters: Construction of C−S Bonds under Transition‐Metal‐Free Conditions

AbstractA convenient and effective method for the construction of C−S bond under transition‐metal free conditions was reported. Thus, diverse thiocarbamide esters could be smoothly obtained in moderate to good yields from inexpensive and readily available starting materials, such as phenyl isothiocyanate, alcohols and benzyl halides. This method features mild conditions, easy operation, short reaction time and broad substrate scopes, providing a facile and convenient strategy for the preparation of some compounds with potential biological activities.

Adding value to strawberry agro-industrial by-products through ultraviolet A-induced biofortification of antioxidant and anti-inflammatory phenolic compounds.

The revalorization of agro-industrial by-products by applying ultraviolet A (UVA) radiation to biofortify with phenolic compounds has been studied in recent times, showing improvements in the individual and total phenolic content and their bioactivity. Therefore, the main aim of this work was to optimize the biofortification process of phenolic compounds by UVA radiation to strawberry agro-industrial by-products (RF). Moreover, the effect of UVA radiation on the potential biological activity of the phenolics accumulated in RF due to the treatment was also determined. The assays followed a factorial design with three variables at three levels: UVA dose (LOW, MEDIUM, and HIGH), storage temperature (5, 10, and 15°C), and storage time (0, 24, 48, and 72 h). At each experimental condition, phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO) enzymatic activities, total phenolic compound content (TPC), phenolics profile (TPCHPLC), and agrimoniin content (AGN) were evaluated; and the optimal UVA dose, storage time, and temperature were determined. In vitro bioaccessibility of the accumulated phenolic compound was studied on RF tissue treated with UVA at optimal process conditions. The digested extracts were tested for antiproliferative activity in colorectal cancer cells, cellular antioxidant capacity, and anti-inflammatory activity. The results showed that applying UVA-HIGH (86.4 KJ/m2) treatment and storing the tissue for 46 h at 15°C increased PAL activity (260%), phenolic content (240%), and AGN (300%). The biofortification process improves the bioaccessibility of the main phenolic compound of RF by 9.8 to 25%. The digested optimum extract showed an IC50 for HT29 and Caco-2 cells of 2.73 and 5.43 μg/mL, respectively, and presented 60% cellular antioxidant capacity and 30% inhibition of NOX production. The RF treated with UVA is an excellent source of phenolic compounds; specifically, ellagitannins and the UVA radiation proved to be efficient in biofortify RF, significantly improving the phenolic compounds content and their bioactive properties with adequate bioaccessibility, adding value to the strawberry agro-industrial by-products.

Evaluation of pharmacological activities and active components in Tremella aurantialba by instrumental and virtual analyses.

As a kind of medicinal and edible homologous fungus, there is a lack of data on the medicinal value of Tremella aurantialba. In this study, ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF/MS) was used to screen the chemical components in T. aurantialba. Then, network pharmacology was used to reveal the potential biological activities, active compounds, and therapeutic targets of T. aurantialba. Finally, the potential binding sites of the active compounds of T. aurantialba and key targets were studied by molecular docking. Results showed that 135 chemical components in T. aurantialba, especially linoleic acid, and linolenic acid have significant biological activities in neuroprotective, anticancer, immune, hypoglycemic, and cardiovascular aspects. The existence of these bioactive natural products in T. aurantialba is consistent with the traditional use of T. aurantialba. Moreover, the five diseases have comorbidity molecular mechanisms and therapeutic targets. The molecular docking showed that linolenic acid, adenosine, and vitamin D2 had higher binding energy with RXRA, MAPK1, and JUN, respectively. This study is the first to systematically identify chemical components in T. aurantialba and successfully predict its bioactivity, key active compounds, and drug targets, providing a reliable novel strategy for future research on the bioactivity development and utilization of T. aurantialba.

Green-Based Approach to Synthesize Silver Nanoparticles Using the Fungal Endophyte Penicillium oxalicum and Their Antimicrobial, Antioxidant, and In Vitro Anticancer Potential.

A green-based approach for the synthesis of silver nanoparticles has gained tremendous attention in biomedical applications. Fungal endophytes have been recognized as a remarkable biological source for the synthesis of potential nanodrugs. The present study focuses on the fabrication of silver nanoparticles using the fungal endophyte Penicillium oxalicum (POAgNPs) associated with the leaf of the Amoora rohituka plant. Sharp UV-visible spectra at 420 nm appeared due to the surface plasmon resonance of POAgNPs and the reduction of silver salt. FT-IR analysis revealed the presence of functional groups of bioactive compounds of P. oxalicum responsible for the reduction of silver salt and validated the synthesis of POAgNPs. A high degree of crystallinity was revealed through XRD analysis, and microscopy-based characterizations such as AFM, TEM, and FESEM showed uniformly distributed, and spherically shaped nanoparticles. Furthermore, POAgNPs showed a potential inhibitory effect against bacterial and fungal strains of pathogenic nature. POAgNPs also exhibited potential antioxidant activity against the synthetically generated free radicals such as DPPH, superoxide, hydroxyl, and nitric oxide with EC50 values of 9.034 ± 0.449, 56.378 ± 1.137, 34.094 ± 1.944, and 61.219 ± 0.69 μg/mL, respectively. Moreover, POAgNPs exhibited cytotoxic potential against the breast cancer cell lines, MDA-MB-231 and MCF-7 with IC50 values of 20.080 ± 0.761 and 40.038 ± 1.022 μg/mL, respectively. POAgNPs showed anticancer potential through inhibition of wound closure and by altering the nuclear morphology of MDA-MB-231 and MCF-7 cells. Further anticancer activity revealed that POAgNPs induced apoptosis in MDA-MB-231 and MCF-7 cells by differential expression of genes related to apoptosis, tumor suppression, and cell cycle arrest and increased the level of Caspase-3. The novel study showed that P. oxalicum-mediated silver nanoparticles exhibit potential biological activity, which can be exploited as nanodrugs in clinical applications.

SYNTHESIS, CHARACTERIZATION AND ANTI-TUBERCULAR ACTIVITY OF 1, 3- THIAZINE DERIVATIVES

Due to their potent biological activity, heterocyclic compounds have been used extensively in pharmacological research.Researchers' interest has been drawn to the heterocyclic molecule thiazine, which has four carbon atoms, one nitrogenatom, and one sulphur atom. It has interesting pharmacological properties. In the present work, a series of new thiazinederivatives were synthesized from substituted chalcone derivatives. Starting material was acetophenone derivatives. Thestructures of these compounds were confirmed by IR, NMR, Mass and elemental analysis. The synthesized compounds(IVa-IVe) were screened for anti-tuberculosis activity against Mycobacterium tuberculosis and the results show that some ofthese derivatives possess good activity against Mycobacterium tuberculosis.

Eco-friendly Syntheses of Bioactive Iodoflavone from Chalcone under Microwave Condition for the Screening of Their Antibacterial Activity

Flavonoids possess potent biological activity due to the presence of heterocyclic ring moiety and have been established as one of the biologically important scaffolds. Iodinated-flavones were synthesized from corresponding chalcones. Both conventional and microwave techniques were used to synthesize these compounds to make a comparative study. A significant reduction in reaction time and increase in % yield has been observed in the microwave technique. Spectroscopic techniques such as UV, IR, and NMR have been used to confirm the structures of all compounds. Agar disk diffusion method was employed to evaluate the antibacterial activities of the newly synthesized compounds against some bacterial strains such as Salmonella typhi, Streptococcus spp, Shigella spp. etc. Iodoflavone was found to be higher potency against most the bacteria, while others were moderately active. J. of Sci. and Tech. Res. 3(1): 69-76, 2021

Biosynthesis of Levan by Halomonas elongata 153B: Optimization for Enhanced Production and Potential Biological Activities for Pharmaceutical Field

Biosynthesis of Levan by Halomonas elongata 153B: Optimization for Enhanced Production and Potential Biological Activities for Pharmaceutical Field

Total flavonoid content and analgesic activity of ethyl acetate fraction of Indian jujube (Ziziphus mauritiana lamk) leaves

Indian jujube leaves (Ziziphus mauritiana Lamk) have various potential biological activity. Previous studies have proven that ethanol extracted from Indian jujube (Z. mauritiana Lamk) leaves had analgesic activity. This study aimed to determine the total flavonoid content and analgesic activity of the ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves ethanol extract. Tested fraction was prepared by maceration using ethanol as solvent followed by fractionation using ethyl acetate. Identification of active compounds was performed using color reaction, and determination of total flavonoids by colorimetric method. Ethyl acetate fraction was further tested for its analgesic activity using the writhing method. Swiss-Webster white male mice were divided into 5 treatment groups consisting of negative control contained sodium-CMC, positive control contained sodium diclofenac, while groups 3-5 were given ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves with doses of 50, 150, and 200 mg/kg BW. All interventions were administered as a single oral dose. 0.7% (w/v) acetic acid was used as a pain inducer. Analgesic activity was measured by calculating the percentage of writhing protection as a measure of the analgesic effect produced by each intervention. The data were analyzed using one-way Anova to compare analgesic activity between treatment groups. Results showed that ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves at doses of 50, 150, and 200 mg/kg BW gave an effect that was not significantly different from sodium diclofenac as positive control. The ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves ethanol extract contained flavonoids, saponin, and tannins with total flavonoid content of 22.41 ± 1.16% w/w. It can be concluded that the ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves have analgesic activity in mice.

Capture-SELEX of DNA Aptamers for Estradiol Specifically and Estrogenic Compounds Collectively.

Estrogenic compounds such as estrone (E1), 17β-estradiol (E2), and 17α-ethynylestradiol (EE2) are serious environmental contaminants due to their potent biological activities. At least six selections were previously reported to obtain DNA aptamers for E2, highlighting its environmental importance. A careful analysis revealed that the previous aptamers either are too long or do not bind optimally. Herein, a series of new aptamers were obtained from the capture-SELEX method with dissociation constants down to 30 nM as determined by isothermal titration calorimetry (ITC). Two aptamers were converted to structure-switching fluorescent biosensors, which achieved a limit of detection down to 3.3 and 9.1 nM E2, respectively. One aptamer showed similar binding affinities to all the three estrogens, while the other aptamer is more selective for E2. Both aptamers required Mg2+ for binding. The proposed sensors were successfully applied in the determination of E2 in wastewater. Moreover, comparisons were made with previous aptamers based on primary sequence alignment and secondary structures. Among previously reported truncated aptamers, ITC showed binding only in one of them. The newly selected aptamers have the combined advantages of small size and high affinities.

Isolation and Characterization of Novel Biological Control Agent Clostridium beijerinckii against Meloidogyne incognita.

One of the most severe soil-borne pathogens in the world is the root-knot nematode (Meloidogyne incognita). Biological control is gaining more importance as environmental awareness increases. Thus, keeping this in mind, a total of 712 bacterial strains were isolated from 117 rhizosphere soil samples and investigated for potential biological control activity against M. incognita. Strain Sneb518 (Clostridium beijerinckii) was identified as having solid biocontrol activity against M. incognita. Sneb518 demonstrated significant inhibition against M. incognita, with J2 mortality reaching 90.73% at 12 h and with eggs hatching at a rate of 6.00% at 24 h, compared to a hatchability level of 29.07% for the control. Additionally, Sneb518 was excellent for enhancing seed germination. The seeds coated with a fermentation broth containing Sneb518 efficiently boosted the germination rate to 88.49%. The effectiveness and stability of C. beijerinckii Sneb518 against M. incognita were then further evaluated in a greenhouse. According to the pot experiment data, Sneb518 considerably (p < 0.05) reduced the number of root galls and egg masses on roots and also significantly (p < 0.05) increased tomato plant growth. C. beijerinckii Sneb518-treated tomato seedlings exhibited 50.26% biocontrol effectiveness compared to the control group. Our results demonstrate that C. beijerinckii Sneb518 can be a potential biological control agent against root-knot nematode disease and a biomass enhancer. This research will give new options for the sustainable control of root-knot nematode disease in tomatoes and other host plants.

Antioxidant, anti-inflammatory and anti-acne activities of stingless bee (Tetragonula biroi) propolis

We collected stingless bee propolis Tetragonula biroi in order to find materials for medicine and cosmetics applications from tropical rainforest resources. Even though this bee has some biological functions including a cancer cell line, hair growth promotion, asthma remedy, α-glucosidase enzyme inhibition, and antiviral action, the investigation on anti-acne has not been reported yet. This study was to focus on propolis Tetragonula biroi extracts and leads us to isolate active compounds for antioxidant, anti-inflammatory, and anti-acne. We used methanol to obtain the extract from this propolis and assayed it with antioxidants, anti-inflammation, and anti-acne. The extract showed strong activity in antioxidants by DPPH radical scavenging activity (82.31% in 6.25 μg/ml). Via a column chromatography and Reveleris PREP purification system, we isolated 3’–O–methyldiplacone, nymphaeol A, and 5,7,3′,4′–tetrahydroxy–6–geranyl flavonol. These compounds showed potential biological activity with IC50 for antioxidant 6.33, 15.49, 17.32 μM; and antiinflammatory 121.54, 121.20, 117.31 μM. The isolated compounds showed anti-acne properties with properties 0.00, 14.11, and 13.78 mm for the inhibition zone (at a concentration of 1 μg/well), respectively. The results indicated that the propolis extract of Tetragonula biroi has the potential to be developed as a cosmetic agent; however, further work needs to be done to clarify its application.

Recent advances in the biosynthesis of ribosomally synthesized and posttranslationally modified peptides of fungal origin.

Ribosomally synthesized and posttranslationally modified peptides (RiPPs) are growing class of natural products with potent biological activities. Although the core scaffolds of RiPPs are composed of proteinogenic amino acids, remarkable structural diversity is generated through posttranslational modifications (PTMs) of precursor peptides. In addition, ribosomal origin of biosynthetic precursors enables supply of its analogs through genetic approach such as site-directed mutagenesis on corresponding genes. As PTM enzymes often exhibit substrate tolerance, RiPP biosynthetic machineries are considered as efficient tools for generation of unique peptide derivatives. RiPP pathways are distributed among all domains of life and those derived from bacteria and plants have been known for decades. In contrast, fungal RiPPs (F-RiPPs) have fewer examples. Amatoxins and omphalotins are F-RiPPs produced by Basidiomycota fungi. In the biosynthesis of these compounds, macrocyclization by prolyl oligopeptidase homologs and N-methylations of back bone amides have been characterized, respectively. Ustiloxins and related compounds are another group of F-RiPPs with characteristic macrocyclic ethers. UstYa family proteins, which are fungi-specific putative oxidases, have been identified as common proteins involved in PTMs of these compounds. Despite a limited number of characterized examples, recent progress in sequencing of fungal genomes indicated that a number of RiPP pathways are hidden in fungal resources, making F-RiPPs as attractive target for genome mining studies while more detailed understandings of key biosynthetic enzymes are still necessary. This review seeks to describe recent advances on the F-RiPP biosynthesis with slight emphasis on the function of UstYa family proteins.

Unique guanidine-conjugated catechins from the leaves of Alchornea rugosa and their autophagy modulating activity

Natural guanidines, molecules that contain the guanidine moiety, are structurally unique and often exhibit potent biological activities. A phytochemical investigation of the leaves of Alchornea rugosa (Lour.) Müll.Arg. by MS/MS-based molecular networking revealed eight undescribed guanidine-flavanol conjugates named rugonines A–H. The chemical structures of the isolated compounds were comprehensively elucidated by NMR spectroscopy, HRESIMS, and circular dichroism (CD) analysis. All isolated compounds were tested for autophagosome formation in HEK293 cells stably expressing GFP-LC3. The results revealed that compounds rugonines D–G showed potential autophagy inhibitory activity.

Multifunctional bioactive properties of hydrolysates from colocynth (Citrullus colocynthis) seeds derived proteins: Characterization and biological properties

Citrullus colocynthis (Colocynth) has gained a great deal of interest in their applications as indigenous nutraceutical and as a functional food ingredient. The intact colocynth seed protein was enzymatically hydrolyzed using proteolytic enzymes (alcalase, bromelain, and chymotrypsin) at different time intervals of 3, 6, and 9 h. The highest degree of hydrolysis (87.82%) was observed in chymotrypsin derived colocynth seed protein hydrolysates (CSPH) for 9 h. The CSPHs was further investigated through in-vitro assay to explore its potential biological activity such as antioxidant, inhibition of enzymatic marker related to diabetes (DPP-IV, α-glucosidase and α-amylase) and hyperlipidaemia (cholesteryl esterase and pancreatic lipase). Chymotrypsin hydrolysate showed the strongest DPPH (65.7 mM TEAC) and ABTS (525.2 mM TEAC) radical scavenging activity after 6 h of hydrolysis. Moreover, chymotrypsin-treated CSPH for 6 h inhibited cholesteryl esterase (IC50 = 13.68 μg/mL) and pancreatic lipase (IC50 = 14.12 μg/mL) significantly when compared to native protein. Whereas, bromelain and alcalase treated hydrolysate for 6 h effectively inhibited α-glucosidase and α-amylase at an inhibitory concentration of IC50 = 13.27 μg/mL and of IC50 = 17 μg/mL. Overall, the findings indicated that protein hydrolysates exhibited superior biological activity than intact colocynth seed proteins isolate (CSPI) and could be a sustainable source of bioactive peptides.