Introduction
Materials and methods
Chemicals
Identification and preparation of plant material
Preparation of saponin and alkaloid fractions
LC–MS/MS analysis
Determination of in vitro pancreatic lipase inhibitory activity
Determination of in vitro alpha amylase inhibitory activity
Determination of in vitro alpha-glucosidase inhibitory activity
Antiadipogenic properties of the plants on 3T3-L1 adipocytes
Cell culture and differentiation
Cell viability assay
Oil Red O staining of intracellular triglycerides
Lipolysis determination
Molecular docking studies of LCMS/MS identified compounds against α-amylase, lipase and α-glucosidase
Protein structure preparation
Ligand preparation
Molecular docking of phytochemicals with a targeted active site
Statistical analysis
Results
Compounds result of the LCMS/MS analysis
Peak | RT | Mass | Name | Formula | Sub-class | Class |
---|---|---|---|---|---|---|
1 | 4.41 | 170.09 | Furfural diethyl acetal | C9 H14 O3 | Furans | Heterocyclic Compounds |
2 | 7.21 | 370.19 | 10-Hydroxyyohimbine | C21 H26 N2 O4 | Secologanin Tryptamine Alkaloids | Alkaloids |
3 | 7.45 | 228.10 | Depdecin | C11 H16 O5 | Epoxide | Organic heterocyclic compound |
4 | 7.95 | 358.13 | Sweroside | C16 H22 O9 | Iridoid Glycosides | Monoterpenes |
5 | 8.22 | 340.18 | 3-Hydroxyquinidine | C20 H24 N2 O3 | Cinchona Alkaloids | Alkaloids |
6 | 8.47 | 370.19 | 18-Hydroxyyohimbine | C21 H26 N2 O4 | - | Alkaloids |
7 | 8.56 | 352.18 | Ajmalicine | C21 H24 N2 O3 | Monoterpenoid indole alkaloid | Alkaloids |
8 | 8.56 | 384.21 | 16-Methoxy-2,3-dihydro-3-hydroxytabersonine | C22 H28 N2 O4 | Monoterpenoid indole alkaloid | Alkaloids |
9 | 8.63 | 354.19 | Vincamine | C21 H26 N2 O3 | Secologanin Tryptamine Alkaloids | Alkaloids |
10 | 8.74 | 366.19 | Aspidofractine | C22 H26 N2 O3 | Indole alkaloid | Alkaloids |
11 | 8.87 | 366.20 | 16-Methoxytabersonine | C22 H26 N2 O3 | Indole alkaloid | Alkaloids |
12 | 9.03 | 368.18 | Mitraphylline | C21 H24 N2 O4 | Indole alkaloid | Alkaloids |
13 | 9.20 | 384.20 | 16-Methoxy-2,3-dihydro-3-hydroxytabersonine | C22 H28 N2 O4 | Monoterpenoid indole alkaloid | Alkaloids |
14 | 9.39 | 354.19 | Vincamine | C21 H26 N2 O3 | Secologanin Tryptamine Alkaloids | Alkaloids |
15 | 9.47 | 368.17 | Horhammericine | C21 H24 N2 O4 | Indole alkaloid | Alkaloids |
16 | 9.51 | 336.19 | Catharanthine | C21 H24 N2 O2 | Secologanin Tryptamine Alkaloids | Alkaloids |
17 | 9.66 | 336.18 | Catharanthine | C21 H24 N2 O2 | Secologanin Tryptamine Alkaloids | Alkaloids |
18 | 9.74 | 338.17 | 18-carboxy dinor Leukotriene B4 | C18 H26 O6 | Arachidonic Acids | Fatty acids and conjugates |
19 | 9.87 | 404.24 | Estradiol-17-phenylpropionate | C27 H32 O3 | Steroids | - |
20 | 10.26 | 366.19 | Aspidofractine | C22 H26 N2 O3 | Idole alkaloid | Alkaloids |
Peak | RT | Mass | Name | Formula | Sub-class | Class |
---|---|---|---|---|---|---|
1 | 0.89 | 126.00 | 2-Hydroxyethanesulfonate | C2 H6 O4 S | Organosulfonic acids | Organic sulfonic acids and derivatives |
2 | 2.17 | 202.08 | Diethyl Oxalpropionate | C9 H14 O5 | beta-keto acids and derivatives | Keto acids and derivatives |
3 | 3.09 | 186.09 | cis-2-Carboxycyclohexyl-acetic acid | C9 H14 O4 | Dicarboxylic acid | Carboxylic acid |
4 | 7.91 | 404.13 | Gardenoside | C17 H24 O11 | Iridoids | Monoterpenes |
5 | 8.48 | 430.21 | Cinegalline | C23 H30 N2 O6 | Quinolizidine alkaloid | Alkaloid |
6 | 8.73 | 156.08 | C4 H8 N6 O | |||
7 | 8.84 | 204.10 | Diethyl (2R,3R)-2-methyl-3-hydroxysuccinate | C9 H16 O5 | Carboxylic ester | Ester |
8 | 9.22 | 210.05 | Vanilpyruvic acid | C10 H10 O5 | Phenylpyruvic acid derivatives | Carboxylic acid |
9 | 9.49 | 186.09 | cis-2-Carboxycyclohexyl-acetic acid | C9 H14 O4 | Dicarboxylic acid | Carboxylic acid |
10 | 9.78 | 184.08 | Glu-P-2 | C10 H8 N4 | ||
11 | 9.98 | 142.10 | 3-Methyl-2Z-heptenoic acid | C8 H14 O2 | Fatty acids and conjugates | Fatty Acyls |
12 | 10.36 | 186.09 | cis-2-Carboxycyclohexyl-acetic acid | C9 H14 O4 | Dicarboxylic acid | Carboxylic acid |
13 | 11.23 | 272.16 | C14 H24 O5 | |||
14 | 12.07 | 214.12 | alpha-Carboxy-delta-decalactone | C11 H18 O4 | Delta valerolactones | Lactones |
15 | 12.32 | 214.12 | alpha-Carboxy-delta-decalactone | C11 H18 O4 | Delta valerolactones | Lactones |
16 | 15.57 | 458.34 | 18-acetoxy-1α-hydroxyvitamin D3 / 18-acetoxy-1α-hydroxycholecalciferol | C29 H46 O4 | ||
17 | 16.01 | 256.17 | 5-Nonyltetrahydro-2-oxo-3-furancarboxylic acid | C14 H24 O4 | Gamma butyrolactones | Lactones |
18 | 16.56 | 618.39 | 3-O-cis-Coumaroylmaslinic acid | C39 H54 O6 | Triterpenoids | Terpenes |
19 | 16.74 | 648.40 | 3α-O-trans-Feruloyl-2α-hydroxy-12-ursen-28-oic acid | C40 H56 O7 | Triterpenoids | Terpenes |
20 | 16.95 | 278.15 | C17 H18 N4 | |||
21 | 17.28 | 298.25 | C18 H34 O3 | |||
22 | 18.80 | 328.26 | MG(0:0/16:1(9Z)/0:0) | C19 H36 O4 | ||
23 | 19.01 | 456.36 | C27 H52 O3 S | |||
24 | 20.56 | 356.17 | C19 H24 N4 O S | |||
25 | 20.68 | 382.18 | 1,2,10-Trihydroxydihydro-trans-linalyl oxide 7-O-beta-D-glucopyranoside | C16 H30 O10 | O-glycosyl compounds | Carbohydrates and carbohydrate conjugates |
26 | 21.06 | 472.36 | Lucidumol A | C30 H48 O4 | Triterpenoids | Terpenes |
Effect of fractions on α-amylase, lipase and α-glucosidase activities
Alstonia boonei fractions | α-amylase | Pancreatic lipase | α-glucosidase | |||
---|---|---|---|---|---|---|
Inhibition (%) | IC50 (μg/mL) | Inhibition (%) | IC50 (μg/mL) | Inhibition (%) | IC50 (μg/mL) | |
ME | 40.37 ± 2.43 | - | 41.23 ± 1.85 | - | 32.56 ± 5.28 | - |
CFF | 44.64 ± 4.32 | - | 37.98 ± 1.53 | - | 45.92 ± 5.06 | - |
CAF | 74.54 ± 1.40 | 1203 ± 0.364 | 65.42 ± 2.19 | 2.77 ± 0.063 | 42.49 ± 1.63 | - |
CSF | 74.97 ± 3.19 | 206 ± 0.103 | 60.21 ± 4.16 | 25.37 ± 0.062 | 77.95 ± 4.29 | 11.97 ± 0.081 |
Orlistat | NA | NA | 93.18 ± 2.97 | 0.068 ± 0.063 | NA | NA |
Acarbose | 83.18 ± 2.97 | 64.58 ± 0.071 | NA | NA | 76.70 ± 2.63 | 2.84 ± 0.087 |
Antiadipogenic and lipolytic properties of the plants on 3T3-L1 preadipocytes
Molecular docking results
Molecular docking of identified compounds against human α-amylase, lipase and α-glucosidase
Amino acid interaction of top docked compounds with α-amylase, lipase and α-glucosidase
Compounds | Binding Energies (Kcal/mol) | Protein | Hydrogen bonds (Bond distance (Å)) | Hydrophobic Interaction (Bond distance (Å)) | Other interactions (Bond distance (Å)) | |||
---|---|---|---|---|---|---|---|---|
Numbers | Interacting residues | Numbers | Interacting residues | Numbers | Interacting residues | |||
Acarbose | -7.3 | PPA | 21 | Thr 52 Val50 Trp59 Gln63(3) Tyr62 Leu165 His101 Thr163 Ala 198 His305(2) Asp197 Gly306(2) His299 Arg195 Glu233(2) Asp300 | 1 | Trp59 | ||
Estradiol-17-phenylpropionate | -11 | 3 | His299 Arg195 Ala198 | 11 | Tyr62 Ile235 Lys200 Leu162 Trp59 His395 Leu165 Val163 Trp58 Trp62 His101 | 2 | His201 Ile235 | |
3α-O-trans-Feruloyl-2 α -hydroxy-12-ursen-28-oic acid | -10.0 | 3 | His299 Asp197 Gln63 | 6 | Pro54 Trp357 Tyr62 Leu162 His305 Glu233 | 1 | Trp59 | |
Orlistat | -6.7 | PPL | 4 | Phe77 Gly76 His151His263 | 3 | Ile78 Ile209 Phe215 | 1 | Asp79 |
Estradiol-17-phenylpropionate | -10.8 | 1 | Cys256 | 10 | Phe77(2) Tyr114(2) Pro180 Phe215(2) Ala260 His263(2) | none | ||
10-Hydroxyyohimbine | -10.4 | 3 | Phe77 His151Arg256 | 8 | Phe77 Ile78 Pro180 Phe215(2) Ala260(2) Ala259 | none | ||
Acarbose | -7.3 | mG | 23 | Asp68 Tyr71 Gln181 Asp214 Arg443 Arg439 His348 Glu276 Arg212 Gln350 Glu304 Arg312 His239 Asn241 Ser156 Tyr313 Phe157 Gly156 Asn412 Phe310 Pro309 Ser308 Asp408 | 1 | Phe177 | none | |
Estradiol-17-phenylpropionate | -10.1 | 6 | Asp68 Arg212 His358 Asp214 His279 Arg312 | 5 | Pro240 Phe157 Phe177 Phe158 Phe300 | 2 | His239 Asp349 | |
10-Hydroxyyohimbine | -9.9 | 13 | Asp68 Thr215 Arg443 Arg212 His348 Asp214 Asp349 Phe310 Arg312 Pro309 His279 Ser308 Asn241 | 3 | Phe157 Phe177 Phe311 | none |