A UPLC-DAD-Based Bio-Screening Assay for the Evaluation of the Angiotensin Converting Enzyme Inhibitory Potential of Plant Extracts and Compounds: Pyrroquinazoline Alkaloids from Adhatoda vasica as a Case Study
Abstract
:1. Introduction
2. Results and Discussion
2.1. Optimization of UPLC Conditions for the ACE Inhibitory Assay
2.2. Optimization of Angiotensin Converting Enzyme Assay
2.3. ACE Inhibitory Activity of Medicinal Plants and Purified Compounds
2.4. Characterization of Pyrroquinazoline Alkaloids
2.5. Molecular Docking
2.6. ADME/Tox Screening of Compounds
3. Materials and Methods
3.1. Chemicals and Reagents
3.2. UPLC-Based ACE Assay
3.3. Enzymatic Assay
3.4. Method Validation
3.5. Extraction and Isolation of Compounds from Adhatoda vasica
3.6. Characterization of Pyrroquinazoline Alkaloids
3.7. Molecular Docking Screening
3.8. ADME/Tox Screening of Compounds
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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S.No. | HHL Conc. (mM) | Intraday | Inter-Day | ||||
---|---|---|---|---|---|---|---|
Mean Conc. (mM) | RSD % | %Error | Mean Conc. (mM) | RSD % | %Error | ||
1. | 0.8 | 0.8023 | 1.0850 | 0.2929 | 0.8137 | 2.2019 | 1.7230 |
2. | 1.8 | 1.8232 | 0.0841 | 1.2921 | 1.8160 | 0.6852 | 0.8932 |
S.No. | ACE Assay | Intraday | Inter-Day | ||||
---|---|---|---|---|---|---|---|
ACE %Conversion | RSD % | %Error | ACE %Conversion | RSD % | %Error | ||
1. | Control | 70.0940 | 1.8826 | 3.0795 | 69.5008 | 2.7430 | 2.2071 |
2. | Captopril (100 nM) | 28.0923 | 2.6146 | 4.0457 | 27.8100 | 3.0499 | 3.0000 |
3. | Lisinopril (100 nM) | 21.8837 | 3.9631 | 4.0187 | 21.8887 | 3.9927 | 3.9969 |
Inhibitors/Compounds | Binding Free Energy Kcal/mol | Types of Interactions Along with Interacting Amino Acids of ACE |
---|---|---|
Lisinopril | −8.48 kcal/mol | Hydrogen Bonds Glu162, Gln281, His353, Lys511, His513, Tyr520 and Tyr523 Hydrophobic Interactions His 353, Ala354, Ala356, Val380 and His387 |
Captopril | −6.97 kcal/mol | Hydrogen Bonds Gln 281, His353, Glu384, Lys511, Tyr520, Tyr523 and His513 Hydrophobic Interactions His383 and Tyr 523 |
Vasicine | −5.89 kcal/mol | Hydrogen Bonds His513 and Tyr520 |
Vasicinol | −6.40 kcal/mol | Hydrogen Bonds Glu384, His513, and Tyr520 |
Vasicinone | −5.79 kcal/mol | Hydrogen Bonds Asn66, Asn70, Ser355, Trp357 and Lys368 Hydrophobic Interactions Trp357 |
Ligand | Parameters Standards | Lisinopril | Captopril | Vasicinol | Vasicine | Vasicinone |
---|---|---|---|---|---|---|
PubChem ID | – | 5362119 | 44093 | 442934 | 72610 | 442935 |
Mol. Weight | 100.0–600.0 | 405.49 | 217.29 | 204.23 | 188.23 | 202.21 |
LogP | −3.0–6.0 | −1.22 | 0.34 | 0.09 | 0.44 | 0.52 |
HBA | ≤12 | 8 | 4 | 4 | 3 | 4 |
HBD | ≤7 | 5 | 1 | 2 | 1 | 1 |
Tpsa | ≤11 | 144.82 | 99.24 | 57.67 | 37.44 | 55.12 |
RotatableB | ≤11 | 12 | 3 | 0 | 0 | 0 |
RigidB | ≤30 | 15 | 8 | 15 | 15 | 16 |
Rings | ≤6 | 2 | 1 | 1 | 1 | 1 |
Max Size Ring | ≤18 | 6 | 5 | 13 | 13 | 13 |
Carbon Atoms | 3–35 | 21 | 9 | 11 | 11 | 11 |
Hetero Atoms | 1–15 | 8 | 5 | 4 | 3 | 4 |
Ratio H/C | 0.1–1.1 | 0.38 | 0.56 | 0.36 | 0.27 | 0.36 |
NumCharges | ≤4 | 4 | 1 | 1 | 1 | 0 |
Total Charge | −8 | 0 | −1 | |||
Lipinski Violation | – | 0 | 0 | 0 | ||
Solubility(mg/L) | 170317.77 | 65224.97 | 54156.75 | 43933.32 | 36306.82 | |
Solubility Forecast Index | Good Solubility | Good Solubility | Good Solubility | Good Solubility | Good Solubility | |
Oral Bioavailability (VEBER) | Low | Good | ||||
Oral Bioavailability (EGAN) | ||||||
StereoCenters | 3 | 2 | 1 | |||
Result | Accepted |
Position | Compound 1 | Compound 2 | Compound 3 | |||
---|---|---|---|---|---|---|
δH (mult., J in Hz) | δC13 | δH (mult., J in Hz) | δC13 | δH (mult., J in Hz) | δC13 | |
1 | 3.75–3.61 (m) | 53.02 | 3.33–3.45 (m) | 47.52 | 4.29–4.02 (m) | 44.58 |
2 | 2.12–2.06 (m) | 31.76 | 2.41–2.18 (m) | 30.78 | 2.66–2.18 (m) | 31.08 |
3 | 5.11 (t, 8.0) | 73.46 | 4.66 (t, 6.4) | 72.54 | 5.12 (t, 7.8) | 73.26 |
3a | --- | 163.77 | --- | 164.76 | --- | 161.84 |
4a | 126.22 | 132.43 | 150.47 | |||
5 | 6.98 (d, 8.8) | 121.38 | 7.18 (d, 8.1) | 130.47 | 8.24 (dd, 8.2, 1.2) | 135.67 |
6 | 6.75 (dd, 8.8, 2.4) | 118.54 | 7.33 (dt, 8.0, 4.0) | 128.35 | 7.84 (dt, 8.3, 1.4) | 128.00 |
7 | --- | 157.75 | 7.25 (dt, 8.0, 4.0) | 128.26 | 7.55 (dt, 8.1, 1.2) | 127.16 |
8 | 6.62 (d, 2.8) | 116.23 | 7.13 (d, 8.0) | 118.42 | 7.73 (d, 7.8) | 128.01 |
8a | --- | 121.18 | --- | 118.24 | --- | 121.91 |
9 | 4.81–4.77, 2H (d, 16.0) | 48.89 | 5.08 (d, 15) | 51.95 | 162.59 |
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Tehreem, S.; Rahman, S.; Bhatti, M.S.; Uddin, R.; Khan, M.N.; Tauseef, S.; El-Seedi, H.R.; Bin Muhsinah, A.; Uddin, J.; Musharraf, S.G. A UPLC-DAD-Based Bio-Screening Assay for the Evaluation of the Angiotensin Converting Enzyme Inhibitory Potential of Plant Extracts and Compounds: Pyrroquinazoline Alkaloids from Adhatoda vasica as a Case Study. Molecules 2021, 26, 6971. https://doi.org/10.3390/molecules26226971
Tehreem S, Rahman S, Bhatti MS, Uddin R, Khan MN, Tauseef S, El-Seedi HR, Bin Muhsinah A, Uddin J, Musharraf SG. A UPLC-DAD-Based Bio-Screening Assay for the Evaluation of the Angiotensin Converting Enzyme Inhibitory Potential of Plant Extracts and Compounds: Pyrroquinazoline Alkaloids from Adhatoda vasica as a Case Study. Molecules. 2021; 26(22):6971. https://doi.org/10.3390/molecules26226971
Chicago/Turabian StyleTehreem, Syeda, Saeedur Rahman, Muhammad Salman Bhatti, Reaz Uddin, Muhammad Noman Khan, Saba Tauseef, Hesham R. El-Seedi, Abdullatif Bin Muhsinah, Jalal Uddin, and Syed Ghulam Musharraf. 2021. "A UPLC-DAD-Based Bio-Screening Assay for the Evaluation of the Angiotensin Converting Enzyme Inhibitory Potential of Plant Extracts and Compounds: Pyrroquinazoline Alkaloids from Adhatoda vasica as a Case Study" Molecules 26, no. 22: 6971. https://doi.org/10.3390/molecules26226971