*3.11. Statistical Analysis*

Data from α-amylase, α-glucosidase and anti-inflammatory assays were compared by ANOVA using GraphPad InStat software (San Diego, CA, USA). Differences among means were determined by the least significant difference Tukey's post hoc test, with significance defined at *p* < 0.05 (*n* = 3 to 9).

### **4. Conclusions**

The present study reports the purification and identification of a new 23-*O*-β-Dallopyranosyl-5β,19-epoxycucurbitane-6,24-diene triterpene (charantoside XV, **6**) along with five known cucurbitane-type triterpene glycosides and one sterol from crude acetone extract of an Indian cultivar of *Momordica charantia* L. var. *charantia*. Most of the isolated compounds downregulated the expression of pro-inflammatory *IL-6*, *TNF-α*, and *iNOS*, and mitochondrial marker *COX-2*. The most noteworthy activity was found in the downregulation of *IL-6* expression. With these findings, we evidence that the cucurbitane-type triterpene compounds could be responsible for the decreasing of the expression of defined markers. Further studies will incorporate western blot analysis to evaluate the induction of proteins related to inflammation in response to bitter melon triterpenes. Compounds also showed an antidiabetic potential by inhibition of the α-amylase and α-glucosidase carbolytic enzymes. Compounds docked as inhibitors of porcine pancreatic α-amylase complex showed binding energies from −14.53 to −8.94 kcal/mol and inhibition constants from 22.5 pM to 278.19 nM while when docked with α-glucosidase crystalline structure of isomaltase from *S. cerevisiae* binding energies ranged from −12.43 to −8.23 with inhibition constants from 706.7 pM to 929.04 nM.

**Supplementary Materials:** The following are available online.Scheme S1: Flow chart of the isolation process of compounds **1**–**7**, Figure S1. Chemical structure and LC-HRESIMS of compound **6**, Figure S2. 1H NMR spectrum of compound **6**, Figure S3. 13C NMR spectrum of compound **6**, Figure S4. DEPT-135 spectrum of compound **6**, Figure S5. DQCOSY spectrum of compound **6**, Figure S6. HMQC spectrum of compound **6**, Figure S7. HMBC spectrum of compound **6**, Figure S8. TOCSY spectrum of compound **6**, Figure S9. NOESY spectrum of compound **6**, Figure S10. 1H NMR spectrum of compound **1**, Figure S11. 13C NMR and DEPT 135 spectra of compound **1**, Figure S12. 1H NMR spectrum of compound **2**, Figure S13. 13C NMR and DEPT 135 spectra of compound **2**, Figure S14. 1H NMR spectrum of compound **3**, Figure S15. 13C NMR and DEPT 135 spectra of compound **3**, Figure S16. 1H NMR spectrum of compound **4**, Figure S17. 13C NMR and DEPT 135 spectra of compound **4**, Figure S18. 1H NMR spectrum of compound **5**, Figure S19. 13C NMR and DEPT 135 spectra of compound **5**, Figure S20. 1H NMR spectrum of compound **2**, Figure S21. 13C NMR and DEPT 135 spectra of compound **7**, Figure S22. 2D ligand-protein interactions of purified compounds with the α-amylase enzyme, Karaviloside VI (A), Karaviloside VIII (B), Momordicoside L (C), Momordicoside A (D), Charantoside XIV (E), and Kuguaglycoside C (F). Legends below the figure show the different types of bonding between compounds and the enzymatic pocket of α-amylase, Figure S23. 2D ligand-protein interactions of purified compounds with the isomaltase enzyme, Karaviloside VI (A), Karaviloside VIII (B), Momordicoside L (C), Momordicoside A (D), Charantoside XV (E), and Kuguaglycoside C (F). Legends below the figure show the different types of bonding between compounds and the enzymatic pocket of isomaltase, Table S1: Protein-ligand interactions and docking scores of isolated compounds (**2**–**7**) in α-amylase and isomaltase obtained using AutoDock 4.2.

**Author Contributions:** W.H.P., S.R.S., G.K.J., and B.S.P. conceived and designed the study. W.H.P. purified and elucidated compound structures; S.R.S., J.L.P., D.M.K., and Y.S. conducted the bioassays. All authors shared analyzing the data and writing the manuscript. All authors have read and agreed to the published version of the manuscript.

**Funding:** This study was supported by the United States Department of Agriculture-NIFA-SCRI-2017-51181-26834 through the National Center of Excellence for Melon at the Vegetable and Fruit Improvement Center of Texas A&M University.

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** The data presented in this study are available in Supplementary Materials.

**Conflicts of Interest:** The authors declare no conflict of interest.

**Sample Availability:** Samples of compounds **1**–**7** are not available.
