Carpaine

A chronic stress-induced microbiome perturbation, highly enriched in Ruminococcaceae_UCG-014, promotes colorectal cancer growth and metastasis.[Pubmed:38617000]

Int J Med Sci. 2024 Mar 25;21(5):882-895.

Purpose: Mounting evidence indicates that psychological stress adversely affects cancer progression including tumor growth and metastasis. The aim of this study was to investigate the role of chronic stress-induced microbiome perturbation in colorectal cancer (CRC) progression. Methods: Chronic restraint stress (CRS) was used to establish the chronic stress mouse model, behavioral tests were used for the CRS model evaluation. Subcutaneous xenograft model and lung metastasis model were established to investigate the growth and metastasis of CRC promoted by CRS exposure. 16S rRNA gene sequencing and liquid chromatograph-mass spectrometer (LC-MS) were applied to observe the effects of CRS exposure on the alteration of the gut microbiome and microbial metabolites. Bioinformatics analysis and correlation analyses were applied to analyse the changes in the frequency of body mass, tumor volume, inflammatory factors, neuroendocrine hormones and metabolites of the gut microbiota. Results: In this study, we identifed that CRS exposure model was appropriately constructed by achieving expected increases in disease activity index and enhanced depressive-like behaviors. CRS exposure can promote growth and metastasis of CRC. Besides, the data indicated that CRS exposure not only increased the neuro- and immune-inflammation, but also weakened the gut mucosal immunological function. The 16s rRNA gene sequencing data showed that CRS exposure increased the abundance of g_Ruminococcaceae_UCG_014. Furthermore, the LC-MS data indicated that with only 2 exceptions of Carpaine and DG (15:0/20:4(5Z,8Z,11Z,14Z)/0:0), the majority of these 24 metabolites were less abundant in CRS-exposed mice. Bioinformatics analysis and correlation analyses indicated that only Ruminoscoccaceae-UCG-014 was significantly associated with inflammation (IL-6), neurotransmission (5-HT), and microbial metabolism (PS). Conclusion: CRS exposure altered diversity, composition and metabolites of the gut microbiome, with Ruminococcaceae_UCG-014 perturbation consistently correlated to inflammatory responses, suggesting a particular role of this bacterial genus in CRC growth and metastasis.

Foodomics-based metabolites profiling of the Greek yogurt incorporated with unripened papaya peel powder.[Pubmed:38544783]

Food Chem (Oxf). 2024 Mar 11;8:100199.

The food waste of the fruit processing industry is rich in many bio-active components such as polysaccharides, polyphenols, peptides, etc. that own multifaceted health benefits. The valorization of this waste is an intriguing optimization method for various dairy products. Meanwhile, LC-MS-based foodomics has been an emerging approach for the quantitative and qualitative analysis of dairy foods. Untargeted metabolomics has been done of the optimized functional yogurt that contains different levels of unripened papaya peel powder (UPPP) using high-resolution mass spectroscopy for analysis of added bio-active components in the matrix. UPPP comprises a high content of phytochemicals which could give functionality and therapeutic effect to the Greek yogurt. A total of 36 functional metabolites have been identified which have various health-beneficial attributes. Kaempferol, ostruthin, putative Carpaine derivatives, etc. are some of the metabolites of high importance with a wide area coverage in the metabolome. This work highlights the bioactivity of the UPPP and its prebiotic properties added to the functional yogurt as an independent ingredient. The incorporated plant-based ingredients like UPPP can effectively enhance the functional attributes of Greek yogurt, which is a potential synbiotic food.

Carpaine alleviates tendinopathy in mice by promoting the ubiquitin-proteasomal degradation of p65 via targeting the E3 ubiquitin ligase LRSAM1.[Pubmed:38194842]

Phytomedicine. 2024 Feb;124:155323.

BACKGROUND: Currently, there are no specific drugs or targets available for the treatment of tendinopathy. However, inflammation has recently been found to play a pivotal role in tendinopathy progression, thereby identifying it as a potential therapeutic target. Carpaine (CA) exhibits potential anti-inflammatory pharmacological properties and may offer a therapeutic option for tendinopathy. PURPOSE: This study aimed to investigate the effectiveness of CA in addressing tendinopathy and uncovering its underlying mechanisms. METHODS: Herein, the efficacy of CA by local administration in vivo in comparison to the first-line drug indomethacin was evaluated in a mouse collagenase-induced tendinopathy (CIT) model. Furthermore, IL-1beta induced a simulated pathological inflammatory microenvironment in tenocytes to investigate its underlying mechanisms in vitro. Further confirmation experiments were performed by overexpressing or knocking down the selective targets of CA in vivo. RESULTS: The findings demonstrated that CA was dose-dependent in treating tendinopathy and that the high-dose group outperformed the first-line drug indomethacin. Mechanistically, CA selectively bound to and enhanced the activity of the E3 ubiquitin ligase LRSAM1 in tendinopathy. This effect mediated the ubiquitination of p65 at lysine 93, subsequently promoting its proteasomal degradation. As a result, the NF-kappaB pathway was inactivated, leading to a reduction in inflammation of tendinopathy. Consequently, CA effectively mitigated the progression of tendinopathy. Moreover, the LRSAM1 overexpression demonstrated effectiveness in mitigating the tendinopathy progression and its knockdown abolished the therapeutic effects of CA. CONCLUSION: CA attenuates the progression of tendinopathy by promoting the ubiquitin-proteasomal degradation of p65 via increasing the enzyme activity of LRSAM1. The exploration of LRSAM1 has also unveiled a new potential target for treating tendinopathy based on the ubiquitin-proteasomal pathway.

Phytochemical Extract from Carica papaya Leaves and Punica granatum Seeds as Therapy Against Cognitive Impairment in a Murine Model.[Pubmed:37626269]

Mol Neurobiol. 2024 Jan;61(1):450-464.

Mild cognitive impairment (MCI) is defined as inter-stage between normal cognitive aging and major neurocognitive disorder (MND). This state of decay is a crucial factor in treatment to prevent the progression to MND. In this study, our group developed a virtual screening process to evaluate 2568 phytochemical compounds against 5 key proteins associated with MCI and MND. As a result, two potential candidates were identified: Carpaine, found in Carica papaya leaves, and punicalagin, present in Punica granatum. A model of cognitive impairment (CI) was developed in 10-month-old male Sprague Dawley rats by administering aluminum chloride (AlCl(3)) at a dose of 100 mg/kg/day for 30 days. After AlCl(3 )administration period, one of the groups received Carpaine and punicalagin in a phytochemical extract (PE) by oral gavage for 30 days. Novel object recognition test (NOR) was assessed at three different time points (T1 - before CI, T2 - after CI, and T3 - after PE treatment). Glial fibrillary acidic protein (GFAP) and neurofilament light chain (NfL) were identified in the hippocampus of rats at the end of the study period. After administration of AlCl(3), a reduction in discrimination index vs control rats (CI = 0.012 +/- 0.08 vs Control = 0.076 +/- 0.03), was observed. After phytochemical extract treatment, a significant increase in discrimination index values was observed in the PE group 0.4643 +/- 0.13 vs CI group 0.012 +/- 0.08. Additionally, the evaluation of immunohistochemistry showed an increase in GFAP positivity in the hippocampus of the CI groups, while a slight decrease was observed in the PE group. This work addressed a comprehensive methodology that utilized in silico tools to identify phytochemical compounds (Carpaine and punicalagin) as potential candidates for affecting key proteins in CI. The phytochemical extract containing Carpaine and punicalagin resulted in a trend in the decrease of GFAP expression in the hippocampus and improved recognition memory in rats with CI induced by age and AlCl(3) administration.

Molecular Docking of Bacterial Protein Modulators and Pharmacotherapeutics of Carica papaya Leaves as a Promising Therapy for Sepsis: Synchronising In Silico and In Vitro Studies.[Pubmed:36677632]

Molecules. 2023 Jan 6;28(2):574.

Sepsis is a serious health concern globally, which necessitates understanding the root cause of infection for the prevention of proliferation inside the host's body. Phytochemicals present in plants exhibit antibacterial and anti-proliferative properties stipulated for sepsis treatment. The aim of the study was to determine the potential role of Carica papaya leaf extract for sepsis treatment in silico and in vitro. We selected two phytochemical compounds, Carpaine and quercetin, and docked them with bacterial proteins, heat shock protein (PDB ID: 4PO2), surfactant protein D (PDB ID: 1PW9), and lactobacillus bacterial protein (PDB ID: 4MKS) against imipenem and cyclophosphamide. Quercetin showed the strongest interaction with 1PW9 and 4MKS proteins. The leaves were extracted using ethanol, methanol, and water through Soxhlet extraction. Total flavonoid content, DPPH assay, HPTLC, and FTIR were performed. In vitro cytotoxicity of ethanol extract was screened via MTT assay on the J774 cell line. Ethanol extract (EE) possessed the maximum number of phytocomponents, the highest amount of flavonoid content, and the maximum antioxidant activity compared to other extracts. FTIR analysis confirmed the presence of N-H, O-H, C-H, C=O, C=C, and C-Cl functional groups in ethanol extract. Cell viability was highest (100%) at 25 microg/mL of EE. The present study demonstrated that the papaya leaves possessed antibacterial and cytotoxic activity against sepsis infection.

Isolation and characterization of carpaine and dihydroxy derivative of carpaine from leaves of Carica papaya: Development of fast HPLC method and standardization of formulations.[Pubmed:36442395]

J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Dec 15;1213:123533.

Carpaine, a major alkaloid of Carica papaya leaves, is widely studied for its anti-thrombocytopenic activity. The objective of present work was to isolate Carpaine from dried leaves of Carica papaya. Isolation of Carpaine was carried out by solvent extraction techniques followed by column chromatography by gradient elution using dichloromethane: methanol (80:20, v/v) as eluent. Dihydroxy derivative of Carpaine was also identified for the first time from leaves of Carica papaya. Structures of Carpaine and dihydroxy derivative of Carpaine were confirmed by LC-MS/MS followed by IR studies. Fast HPLC method was developed using Sunniest C18 column (150 mm x 4.6 mm) as stationary phase and water (pH 8.5): acetonitrile (60: 40, v/v) as mobile phase at flow rate of 1.0 mL/min. Carpaine was eluted with mean retention time of 6.017 min. The developed HPLC method was used to estimate Carpaine in extract and marketed formulations containing Carica papaya leaves.

Delving into the Therapeutic Potential of Carica papaya Leaf against Thrombocytopenia.[Pubmed:35566112]

Molecules. 2022 Apr 25;27(9):2760.

Thrombocytopenia is a clinical manifestation that refers to the low platelet count, i.e., <150 x 103/muL, of blood, resulting in imbalanced hemostasis, which leads to several fatal complications. The causative factors vary greatly, but, as a consequence, they interfere with platelet production and promote destruction, leading to death. Carica papaya leaf has unique therapeutic and medicinal characteristics against thrombocytopenia, and this is supported by scientific studies. Secondary metabolites and minerals in the leaf, such as Carpaine and quercetin, promote platelet production, inhibit platelet destruction, and maintain platelet membrane through gene expression activity and the ceasing of viral proteases, respectively. This review explores the scientific studies that support the role of papaya leaf in the form of juice, extract, or powder against thrombocytopenia through animal modeling and clinical trials. Phytochemical profiles of C. papaya leaf revealed the presence of flavonoids, alkaloids, phenols, cardiac glycosides, tannins, terpenes, and saponins, which impart therapeutic potential to the leaf. The therapeutic benefits of the leaf include immunomodulatory, antiviral, antidiabetic, anticancer, antimalarial, antiangiogenic, antibacterial, and antioxidant activities. Several conducted scientific research studies have proved the efficacy of C. papaya leaf against thrombocytopenia, expanding the implication of natural sources to eradicate numerous ailments.

Chemical constituents from Carica papaya Linn. leaves as potential cytotoxic, EGFR(wt) and aromatase (CYP19A) inhibitors; a study supported by molecular docking.[Pubmed:35424860]

RSC Adv. 2022 Mar 23;12(15):9154-9162.

The phytochemical investigation of the hydromethanolic extract of Carica papaya Linn. leaves (Caricaceae) resulted in the isolation and characterization of ten compounds, namely; Carpaine (1), methyl gallate (2), loliolide (3), rutin (4), clitorin (5), kaempferol-3-O-neohesperidoside (6), isoquercetin (7), nicotiflorin (8) and isorhamnetin-3-O-beta-d-glucopyranoside (9). The compounds 2, 3, 5-7 and 9 were isolated for the first time from the genus Carica. An in vitro breast cancer cytotoxicity study was evaluated with an MCF-7 cell line using the MTT assay. Methyl gallate and clitorin demonstrated the most potent cytotoxic activities with an IC(50) of 1.11 +/- 0.06 and 2.47 +/- 0.14 muM, respectively. Moreover, methyl gallate and nicotiflorin exhibited potential EGFR(wt) kinase inhibition activities with an IC(50) of 37.3 +/- 1.9 and 41.08 +/- 2.1 nM, respectively, compared with the positive control erlotinib (IC(50) = 35.94 +/- 1.8 nM). On the other hand, clitorin and nicotiflorin displayed the strongest aromatase kinase inhibition activities with an IC(50) of 77.41 +/- 4.53 and 92.84 +/- 5.44 nM, respectively. Clitorin was comparable to the efficacy of the standard drug letrozole (IC(50) = 77.72 +/- 4.55). Additionally, molecular docking simulations of the isolated compounds to EGFR and human placental aromatase cytochrome P450 (CYP19A1) were evaluated. Methyl gallate linked with the EGFR receptor through hydrogen bonding with a pose score of -4.5287 kcal mol(-1) and RMSD value of 1.69 A. Clitorin showed the strongest interaction with aromatase (CYP19A1) for the breast cancer receptor with a posing score of -14.2074 and RMSD value of 1.56 A. Compounds (1-3) possessed a good bioavailability score with a 0.55 value.

Carpaine Promotes Proliferation and Repair of H9c2 Cardiomyocytes after Oxidative Insults.[Pubmed:35215343]

Pharmaceuticals (Basel). 2022 Feb 15;15(2):230.

Carpaine has long been identified as the major alkaloid in Carica papaya leaves that possess muscle relaxant properties. Limited study on the molecular signaling properties of Carpaine urges us to conduct this study that aims to elucidate the mechanism underlying the cardioprotective effect of Carpaine in embryonic cardiomyocytes of the H9c2 cell line. The 50% inhibitory concentration (IC(50)) of Carpaine was first determined using a colorimetric MTT assay to establish the minimum inhibitory concentration for the subsequent test. Using a 1 microM Carpaine treatment, a significant increase in the H9c2 proliferation rate was observed following 24 and 48 h of incubation. A Western blot analysis also revealed that Carpaine promotes the upregulation of the cell cycle marker proteins cyclin D1 and PCNA. Carpaine-induced H9c2 cell proliferation is mediated by the activation of the FAK-ERK1/2 and FAK-AKT signaling pathways. In the setting of ischemia-reperfusion injury (IRI), Carpaine provided a significant protective role to recover the wounded area affected by the hydrogen peroxide (H(2)O(2)) treatment. Furthermore, the oxidative-stress-induced reduction in mitochondrial membrane potential (MMP) and overproduction of reactive oxygen species (ROS) were attenuated by Carpaine treatment. The current study revealed a novel therapeutic potential of Carpaine in promoting in vitro cardiomyocyte proliferation and repair following injury.

Metabolic analysis of unripe papaya (Carica papaya L.) to promote its utilization as a functional food.[Pubmed:33704369]

Biosci Biotechnol Biochem. 2021 Apr 24;85(5):1194-1204.

Papaya (Carica papaya L.) is widely cultivated in tropical and subtropical countries. While ripe fruit is a popular food item globally, the unripe fruit is only consumed in some Asian countries. To promote the utilization of unripe papaya based on the compositional changes of biological active metabolites, we performed liquid chromatography-Orbitrap-mass spectrometry-based analysis to reveal the comprehensive metabolite profile of the peel and pulp of unripe and ripe papaya fruits. The number of peaks annotated as phenolics and aminocarboxylic acids increased in the pulp and peel of ripe fruit, respectively. Putative Carpaine derivatives, known alkaloids with cardiovascular effects, decreased, while carpamic acid derivatives increased in the peel of ripe fruit. Furthermore, the functionality of unripe fruit, the benzyl glucosinolate content, total polyphenol content, and proteolytic activity were detectable after heating and powder processing treatments, suggesting a potential utilization in powdered form as functional material.

Bioassay-Guided Different Extraction Techniques of Carica papaya (Linn.) Leaves on In Vitro Wound-Healing Activities.[Pubmed:31991676]

Molecules. 2020 Jan 24;25(3):517.

Herbal plants are traditionally utilized to treat various illnesses. They contain phytochemicals that can be extracted using conventional methods such as maceration, soxhlet, and boiling, as well as non-conventional methods including ultrasonic, microwave, and others. Carica papaya leaves have been used for the treatment of dengue, fungal, and bacterial infections as well as an ingredient in anti-aging products. Phytochemicals analysis detected the presence of kaempferol, myricetin, Carpaine, pseudoCarpaine, dehydroCarpaine I and II, ferulic acid, caffeic acid, chlorogenic acid, beta-carotene, lycopene, and anthraquinones glycoside. Conventional preparation by boiling and simple maceration is practical, simple, and safe; however, only polar phytochemicals are extracted. The present study aims to investigate the effects of three different non-conventional extraction techniques (ultrasonic-assisted extraction, reflux, and agitation) on C. papaya phytochemical constituents, the antioxidant capacity, and wound-healing activities. Among the three techniques, the reflux technique produced the highest extraction yield (17.86%) with the presence of saponins, flavonoids, coumarins, alkaloids, and phenolic metabolites. The reflux technique also produced the highest 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging with an IC(50) value of 0.236 mg/mL followed by ultrasonic-assisted extraction (UAE) (IC(50): 0.377 mg/mL) and agitation (IC(50): 0.404 mg/mL). At tested concentrations (3.125 microg/mL to 500 microg/mL), all extracts do not exhibit a cytotoxicity effect on the human skin fibroblast, HSF1184. Interestingly, reflux and UAE were active fibroblast proliferators that support 85% (12.5 microg/mL) and 41% (6.25 microg/mL) better cell growth, respectively. Additionally, during the early 24 h of the scratch assay, the migration rate at 12.5 microg/mL was faster for all extracts with 51.8% (reflux), 49.3% (agitation), and 42.5% (UAE) as compared to control (21.87%). At 48 h, proliferated cells covered 78.7% of the scratch area for reflux extract, 63.1% for UAE, 61% for agitation, and 42.6% for control. Additionally, the collagen synthesis was enhanced for 31.6% and 65% after 24 and 48 h of treatment for reflux. An HPLC-MS/MS-QTOF (quadruple time-of-flight) analysis of reflux identified nine phytochemicals, including Carpaine, kaempferol 3-(2G-glucosylrutinoside), kaempferol 3-(2''-rhamnosylgalactoside), 7-rhamnoside, kaempferol 3-rhamnosyl-(1->2)-galactoside-7-rhamnoside, luteolin 7-galactosyl-(1->6)-galactoside, orientin 7-O-rhamnoside, 11-hydroperoxy-12,13-epoxy-9-octadecenoic acid, palmitic amide, and 2-hexaprenyl-6-methoxyphenol. The results suggested that reflux was the best technique as compared to ultrasonic and agitation.

Effect of Andrographis paniculata and Phyllanthus amarus leaf extracts on selected biochemical indices in Drosophila melanogaster model of neurotoxicity.[Pubmed:31899970]

Drug Chem Toxicol. 2022 Jan;45(1):407-416.

Aluminum (Al)-induced toxicity in fruit fly (Drosophila melanogaster) is one of the established models for studying neurotoxicity and neurodegenerative diseases. Alkaloid phytochemicals have been reported to exhibit neuroprotective effects. Therefore, the aim of this study is to determine the effect of alkaloid extracts of Andrographis paniculata and Phyllanthus amarus leaves on Al-induced toxicity in wild type Drosophila melanogaster. The flies were exposed to diets containing 40 mM AlCl(3), and the alkaloid extracts (0.1 and 1 mg/ml). Thereafter, the flies were assessed for learning and memory, as well as locomotor performance 14 days post-treatment. This was followed by homogenizing the flies and the homogenates were assayed for acetylcholinesterase, monoamine oxidase and catalase activities, as well as the malondialdehyde content. The results showed that the alkaloid extracts of both leaves could ameliorate the aluminum-induced behavioral and biochemical impairments in the flies. The HPLC analysis of the alkaloid contents revealed that there is an abundance of Amaryllidaceae alkaloids, caffeine and Carpaine. Thus, alkaloid extracts from these leaves could serve as promising therapeutic candidates for the management of neurodegenerative disease.

In vitro ovicidal and larvicidal activity of Carica papaya seed hexane extract against Strongyloides venezuelensis.[Pubmed:31778389]

Rev Inst Med Trop Sao Paulo. 2019 Nov 25;61:e59.

Strongyloidiasis is a human parasitic disease caused by the helminth Strongyloides stercoralis whose treatment is particularly difficult in immunosuppressed patients due to their low responsiveness to conventional therapy. Carica papaya and its isolated compounds benzyl isothiocyanate, Carpaine and carpasemine are promising compound for the treatment of Strongyloides infections due to their anthelmintic action. This study aims to examine the in vitro ovicidal and larvicidal activity of C. papaya seed hexane extract against Strongyloides venezuelensis, using egg hatching tests and larval motility tests as efficiency markers. The crude extract at the concentrations of 566 - 0.0566 mg/mL or the control with albendazole (0.025 mg/mL) and negative controls (water and PBS) were incubated with an equal volume of egg suspension (+/- 50 specimens) followed by counting of the specimens after 48 h. The same extract and dilutions were added to L3 larvae suspensions (+/-50 specimens) followed by analysis of larvae viability after 24, 48, and 72 h. The extract inhibited egg hatching with high efficiency at concentrations of 56.6 mg/mL (95.74%) and 5.66 mg/mL (92.16%). At the concentrations of 566 mg/mL (100%) and 56.66 mg/mL (97.32%), the extract inhibited larval motility as effectively as ivermectin (0.316 mg/mL; 100%), and more effectively than the other dilutions and the negative controls. The larvicidal effect depended on the extract concentration, but not on the treatment period. Therefore, C. papaya seed hexane extract has anthelmintic potential against S. venezuelensis and is a promising compound for the development of phytotherapies to treat strongyloidiasis.

An In silico approach to identify potential inhibitors against multiple drug targets of Mycobacterium tuberculosis.[Pubmed:31512601]

Int J Mycobacteriol. 2019 Jul-Sep;8(3):252-261.

BACKGROUND: The increasing incidence of multidrug-resistant cases of tuberculosis (TB) and difficulty in treating these cases requires an urgent need to find an effective anti-TB drug. There are many phytochemicals with reported antibacterial and antitubercular activities. Instead of targeting only a single target of Mycobacterium tuberculosis (MTB), this study aims to identify phytochemicals targeting multiple drug targets of MTB through subtractive genomic/proteomic approach followed by in silico screening of phytochemicals with reported anti-TB activity. METHODS: Of 614 essential genes of MTB reported in database of essential genes, 15 gene products were selected using different bioinformatic resources and tools such as PANTHER, Venny, NCBI, and BLAST. RESULTS: Virtual screening analysis of these selected drug targets against identified 148 phytochemicals revealed that amentoflavone, Carpaine, 13'bromo-tiliacorinine, and 2'nortiliacorinine, able to inhibit more than one target of MTB. CONCLUSION: These selected compounds may be proposed as potential inhibitors of MTB and need to be tested in TB culture studies in vitro to assess their anti-TB activity.