Synthesis and Characterization of Natural Product-Based Steroid-Triazole Hybrids via Click Reaction and Evaluation of Their Anticancer and Antibacterial Activities

Abstract

Two highly significant classes of natural and heterocyclic compounds, steroids and triazoles, are pivotal in numerous biological processes, and several steroid conjugates and triazole hybrids exhibiting anticancer and antibacterial activities were reported. In this work, a series of 47 new triazoles based on steroid moieties were synthesized via click chemistry. The effective click reaction of steroid propargyl ethers and steroid acetylenic esters with aryl azides provided quantitative yields of the desired steroid-triazole conjugates. The new compounds were fully characterized based on spectral data, including NMR, HRMS, FT-IR, and single X-ray diffraction analysis. The biological activity of the new steroid-triazole hybrids was evaluated through assessments of their anticancer and antibacterial activities. The screen identified three hits with selective antibacterial activity against the Gram-positive Staphylococcus aureus (MIC 12.5–50 μM; IC₅₀ 8.9–41.5 μM), and one compound was weakly cytotoxic against the triple-negative breast cancer cells MDA-MB-231.

Introduction

Natural products (NPs) are privileged structures and rich source of drugs since ancient times, with many successful examples, including morphine (analgesic), penicillin (antibiotic), digitoxin (cardiac glycoside), and quinine (antimalarial). A recent perspective reported that 45% of compounds in phase III were NPs or hybrids, reflecting the same proportion observed in approved medicines. Therefore, NPs are a validated starting point for drug discovery and hence a source of inspiration to the synthetic community toward the design and synthesis of compounds based on natural product scaffolds.

Natural product-based hybrids are compounds generated via the conjugation of two bioactive natural products (or natural product and synthetic moiety) or fragments to generate a new single entity with improved profiles such as synergistic/enhanced potency, improved delivery, act on multiple targets, and overcome limitations related to poor solubility and stability. − These hybrid molecules leverage the prevalidated biological relevance of the natural product core while expanding the chemical space, leading to the discovery of compounds with unique bioactivities. Hybrid chemical compounds have shown a wide variety of applications, particularly in anticancer and antimicrobial drug discovery. −

Two highly significant classes of natural and heterocyclic compounds, steroids and triazoles, are pivotal in numerous biological processes. Steroids are naturally occurring bioactive organic compounds composed of four fused rings that function mainly as signaling molecules and components of cell membranes. Examples include cholesterol and the sex hormones estradiol and testosterone, in addition to many bioactive steroids isolated from marine and terrestrial sources. ,

Steroid ethers are useful as intermediates for effective inhibitors of human skin testosterone 5α-reductase, as well as anti-HSV-1. , Steroid esters exhibit inhibitory activity for many enzymes such as CYP17α hydroxylase and isozymes of 5α-reductase, inhibition of this enzyme is a therapeutic strategy for the treatment of prostate and breast cancer. , Several hybrids/conjugates based on steroid framework have been developed mainly for anticancer drug discovery. −

On the other hand, although the presence of triazole moiety is rare and does not commonly exist among natural products, it is considered a privileged class of heterocyclic compounds that is widely incorporated into semisynthetic derivatives and hybrid compounds based on natural product scaffolds to enhance their pharmacological properties. Among them, 1,4-disubstituted and 1,4,5-trisubstituted 1,2,3-triazole derivatives are privileged scaffolds in medicinal chemistry as they can easily form noncovalent interactions with enzymes and receptors, by affecting the hydrogen bonding capability, thus conferring diverse biological effects such as anti-influenza, antiepileptic, antimicrobial and anticancer. − Furthermore, due to the favorable properties, medicinal value, and synthetic versatility of 1,2,3-triazole scaffolds, they are incorporated in a number of marketed drugs and are currently used in clinical applications, including rufinamide (anticonvulsant agent), cefatrizine (antibiotic), suvorexant (insomnia), and tazobactam (antibiotic) (Figure ). Among the several 1,2,3-triazole hybrids reported, ,,, the conjugates of the 1,2,3-triazole with the steroid framework are interesting due to the ability of the steroidal core to penetrate the biomembranes and bind to specific hormone receptors, which ultimately can potentially translate to broader biological profiles. ,,,, However, to the best of our knowledge only handful of studies reported the synthesis of steroid-1,2,3-triazole hybrids and evaluated their anticancer activity. ,,,−

1.

Structures of FDA-approved drugs containing 1,2,3-triazole scaffold.

Unlike traditional synthetic methodologies which face significant challenges such as low yields, time consumption, harsh reaction conditions, and the synthesis of multiple byproducts, the Nobel winning prize click chemistry provides an efficient approach toward the synthesis of natural product-based hybrids. Since Kolb, Finn, and Sharpless laid down the foundations of click chemistry in 2001, it became a powerful tool in linking natural product scaffolds with other molecules and forming stable 1,2,3-triazole links. , The reaction conditions have an effect on stereospecificity formation of 1,4- or 1,5-disubstituted-1,2,3-triazoles, hence the emergence of the copper-catalyzed azide–alkyne cycloaddition reaction (CuAAC) as an entry point to 1,4-triazoles. −

In this paper, we report the conjugation of two bioactive moieties, triazoles and steroids, resulting in the synthesis, extensive characterization, and biological evaluation of 47 new steroid-triazole hybrids.

Results and Discussions

Synthesis of Steroid-Triazole Hybrids

Scheme illustrates the synthetic procedure for the synthesis of the target steroid-triazole hybrids 8–56. Initially, the steroid acetylenic compounds 1–7 were synthesized by reacting estrogen or estradiol or cholesterol with propargyl bromide in the presence of K₂CO₃ in refluxing acetone to afford steroid propargyl ethers 1–3 in quantitative yields (69–95%). Furthermore, the reaction of estrogen or dehydroandrosterone or pregnanolone or testosterone with propiolic acid in the presence of N,N’-dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) in dichloromethane (DCM) overnight at room temperature afforded steroid esters 4–7 (12–46%) (Scheme ). The structures of steroid ethers 1–3 , and steroid esters 4–7 ,, were assigned based on the spectral data. The ¹H NMR spectra showed the characteristic absorption signal of an acetylenic proton CH at δ 2.41–3.08 ppm (Table ). Furthermore, mass spectra showed the corresponding peaks of the molecular ions.

1. Synthesis of Steroid-Triazoles 8-56 .

1. Yield, ¹H and ¹³C NMR Chemical Shifts of Characteristic Signals of Compounds 1–7 .

Comp.	Yield (%)	δCH (ppm)	δCH 2  (ppm)	δCOO (ppm)
1	95	2.53 (t)	4.69 (d)	-
2	78	2.52 (t)	4.68 (d)	-
3	69	2.41 (t)	4.21 (d)	-
4	19	3.08 (s)	-	151.4
5	46	2.89 (s)	-	152.2
6	43	2.89 (s)	-	152.3
7	12	2.90 (s)	-	152.9

Azides were synthesized by adding sodium nitrite to aryl amines in hydrochloric acid dropwise at 0–5 °C, then the mixture was stirred for 1 h to generate the diazonium chloride salt. Finally, addition of sodium azide (1.2 equiv) to the solution afforded the desired aryl azides, which were used in click reaction without further purification.

Click reaction: a mixture of steroid propargyl ethers 1–3 or acetylenic esters 4–7 and aryl azides were stirred in the presence of catalytic amounts of copper sulfate and sodium ascorbate in a solution of butanol/water (7:3). The heterogeneous mixture was stirred under reflux overnight. The crude products were then purified by flash chromatography to afford steroid-triazole conjugates 8–56 in high yields (62–95%) (Scheme and Table ). The structures of steroid-triazole hybrids 8–56 were confirmed based on analysis of their respective spectral data. The ¹H NMR spectra showed the characteristic absorption signal of the triazoles proton CH-N at low-field δ 7.55–9.26 ppm (Table ). Furthermore, mass spectra showed the corresponding peaks of the molecular ions. Single crystals X-ray diffraction for compounds 19, 38, and 47 were investigated (Figure ). Summary on the nature and various crystallographic parameters are provided in the Supporting Information.

2. Yield, ¹H and ¹³C NMR Chemical Shifts of Characteristic Signals of Compounds 8–56 .

Comp.	Yield (%)	δCH-N (ppm)	δCH 2  (ppm)	δCOO (ppm)
8	87	8.93	5.19	-
9	88	8.05	5.29	-
10	92	8.05	5.29	-
11	81	8.94	5.17	-
12	89	8.95	5.17	-
13	83	9.14	5.20	-
14	82	9.08	5.21	-
15	86	9.04	5.20	-
16	65	9.08	5.21	-
17	83	8.59	5.14	-
18	90	8.05	5.29	-
19	78	7.55	5.19	-
20	75	8.84	5.16	-
21	81	7.64	5.31	-
22	73	7.79	5.30	-
23	62	9.26	5.22	-
24	91	8.63	5.12	-
25	94	8.61	5.12	-
26	78	8.63	5.16	-
27	83	8.33	5.28	-
28	87	8.06	5.29	-
29	90	8.07	5.29	-
30	90	8.36	5.27	-
31	85	8.08	5.29	-
32	82	8.29	5.31	-
33	80	8.26	5.30	-
34	79	9.03	5.19	-
35	63	9.07	5.19	-
36	90	8.58	5.13	-
37	84	8.03	5.31	-
38	71	7.56	5.20	-
39	78	8.19	5.27	-
40	77	7.64	5.31	-
41	76	7.83	5.27	-
42	88	8.25	5.27	-
43	89	9.04	5.12	-
44	92	8.61	5.11	-
45	86	8.62	5.14	-
46	69	8.06	5.12	-
47	91	8.68	-	159.4
48	65	8.79	-	159.1
49	85	8.55	-	162.5
50	87	8.61	-	160.5
51	90	8.54	-	160.5
52	65	8.59	-	164.5
53	95	8.40	-	160.8
54	78	8.59	-	160.9
55	84	8.58	-	160.7
56	67	8.44	-	160.8

2.

Crystal structure of compounds 19, 38, and 47 obtained from single crystal diffraction. Color code: gray-carbon; red-oxygen; blue-nitrogen; black-hydrogen.

Biological Investigations of Steroid-Triazole Hybrids

Steroid conjugates, specially steroid-triazole hybrids, have high antiproliferative potential and their anticancer activity has been extensively evaluated. ,− , This had prompted us to synthesize a series of 49 steroid-triazoles and investigate their anticancer activity, as only compounds 19 and 38 have been reported previously as antiproliferative agents. ,

The anticancer activity of the steroid-triazole hybrids was investigated by screening the library composed of 49 steroid-triazole hybrids, among which 47 compounds were new, against the highly aggressive triple negative breast cancer MDA-MB-231 cells. One hit (42) was identified from the screen exhibiting 44% cell viability at 100 μM following treatment for 48 h (Figure ). The IC₅₀ value of this hit compound could not be reached within the tested range. In comparison with literature, the previously reported antiproliferative activity of steroid-triazole hybrids was based on MTT assay following 72 h incubation period with test compounds. As the library did not demonstrate any cytotoxicity against MDA-MB-231 cells and instead of screening against other cancer cell lines, we aimed at assessing the antibacterial activity as it has not been previously investigated for the reported steroid-triazole hybrids.

3.

Heatmap summarizing the primary biological screening results of the steroid-triazoles library. The rows correspond to the screened compounds while the columns correspond to the primary biological activity screens against S. aureus (Gram-positive pathogen), E. coli (Gram-negative pathogen), and MDA-MB-231 triple negative breast cancer cells and final concentrations of test compounds (100, 10, and 1 μM). *Kanamycin was used as a positive control in the antibacterial screen and its final concentrations tested were 100, 10, and 1 μg/mL.

The library of steroid-triazole hybrids was further screened against Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria at three concentrations (100, 10, and 1 μM) (Figure ). Three hits (46, 47, and 52) were identified from the primary screen to be active against S. aureus, with 52 being the most potent compound. However, none of the screened compounds showed any inhibitory activity against E. coli (Figure ).

The three hits identified from the primary antibacterial screen (Figures and A) along with the positive control, kanamycin, were validated and further tested against S. aureus in a dose–response format to determine their minimum inhibitory concentrations (MIC) and IC₅₀ values (Figures B and D). Compound 46 exhibited an MIC of 50 μM (IC₅₀ 41.5 μM), whereas 52, the most potent analogue from the primary screen, had an MIC of 12.5 μM (IC₅₀ 8.9 μM). We further investigated the effect of our three active compounds on bacterial biofilm inhibition using the same strain (S. aureus). The compounds were able to inhibit the biofilm and exhibited the same MIC values (Figure C).

4.

MIC assay of hit compounds against S. aureus. (A) Structures of hit compounds. (B) Evaluation of planktonic eradication. (C) Evaluation of biofilm eradication. (D) Dose–response curves. Kanamycin was used as a positive control. Test range: 0.1–100 μM for compounds 46, 47, and 52; and 0.1–100 μg/mL for kanamycin (2-fold serial dilution). The experiment was carried out in triplicate n = 3.

Although no clear structure–activity relationship (SAR) was concluded from the library as a whole, comparing the structures of the three hits (46, 47, and 52) may provide an insight into the key structural features that are important for the antibacterial activity. Adding the trifluoromethyl group to the aryl group in 46 and 52 improved the activity which might be due increase in lipophilicity and penetration through cell membrane. Furthermore, converting the aromatic A ring of the steroid framework in 47 to aliphatic as in 46 and 52 improved the activity which might be due modulation of receptor specificity. , The incorporation of a side chain on C17 of the steroid’s D ring as in 46 and 52 may have improved the activity which might be due modulation of receptor specificity, metabolic and physical properties such as lipophilicity and cell membrane penetration, depending on the structure and length of the side chain. The linker type seems to influence the activity as replacement of the ether linker in 46 with an ester in 52 significantly improved the antibacterial activity.

To the best of our knowledge, this data provides the first report of the antibacterial activity of natural product-based steroid-triazole hybrids. The Gram-positive pathogen S. aureus is the leading cause of morbidity and mortality resulting from several community and hospital acquired infections ranging from moderately severe skin infections to fatal pneumonia and sepsis. , Considering the emergence and prevalence of multidrug-resistant strains such methicillin-resistant S. aureus (MRSA), the discovery and development of novel antibiotics is crucial. Furthermore, biofilm-related infections possess a significant clinical challenge due to their inherent resistance to conventional antibiotics and host immune responses and are one of the leading causes for persistent infections. The robust nature of the biofilm matrix acts as a formidable barrier, necessitating effective strategies for biofilm inhibition to improve patient outcomes. Therefore, developing novel approaches that specifically target and disrupt biofilm formation is critical for revolutionizing the treatment of chronic and persistent infections. Taken together, the three hits (46, 47, and 52) identified from our initial antibacterial screen are the starting point for further optimizations aimed at the discovery of more potent antibacterials targeting S. aureus infections.

Conclusion

A library of 47 new natural product-based steroid-triazole hybrids were synthesized via click reaction. All compounds have been structurally characterized from their respective NMR and HRMS spectral data along with single X-ray diffraction analysis (for compounds 19, 38, and 47). Screening the library of the newly synthesized steroid-triazoles against two strains of bacteria (S. aureus and E. coli) identified three hit compounds (46, 47, and 52) with antibacterial activity selectively targeting S. aureus. Validation of the hits identified compound 52 as the best candidate, demonstrating moderate antibacterial and biofilm killing activities (MIC of 12.5 μM, IC₅₀ 8.9 μM). The anticancer screen identified one hit (42) against the highly aggressive triple negative breast cancer MDA-MB-231 cells, exhibiting 44% cell viability at 100 μM following 48 h incubation. The current data represent initial screening; hence, the identified hits are just a starting point for further optimizations. In this library, although many steroid-triazole hybrids did not show significant antibacterial and anticancer activities, further screening against other targets is required to explore other potential activities and fully investigate their therapeutic potential.

Experimental Section

General

All reactions were carried out under nitrogen atmosphere unless otherwise noted. All analyses were determined in the Research Sector Projects Unit (RSPU) at the Faculty of Science, Kuwait University. Thin layer chromatography (TLC) was performed using Polygram SIL G UV254 TLC plates, and visualization was carried out by ultraviolet lights at 254 and 350 nm. Column chromatography was performed using Merck silica gel 60 of mesh sizes 0.040–0.063 mm. ¹H and ¹³C NMR spectra were recorded using Bruker DPX 600 at 400 and 600 MHz. Single-crystal data collection was made on a Bruker X8 Prospector diffractometer. Melting points were determined via differential scanning calorimetry (DSC) analyses on Shimadzu DSC-50.

Crystal Structure Analysis

Single crystals of 19, 38 and 47 were grown by the slow solvent evaporation method. Single-crystal data collection was made on the Bruker X8 Prospector diffractometer by Cu–Ka.

Materials

Estrogen, β-estradiol, trans-dehydroandrosterone (5-androsten-3β-ol-17-one), pregnanolone, and testosterone (17β-hydroxy-3-oxo-4-androstene) are commercially available. Azides required for this study were synthesized using commercial amines.

General Procedures: Synthesis of 1–56

Synthesis of Steroid Propargyl Ethers 1–3

Estrone/estradiol/cholesterol (3 mmol) was added to (4.5 mmol) propargyl bromide and (2–4 equiv) K₂CO₃ in 100 mL acetone. The reaction mixture was refluxed at 80̊C until reaction completion based on TLC monitoring. On completion, the reaction mixture was cooled down to room temperature and the solvent was removed under reduced pressure. The residue was dissolved in distilled water and then extraction was done using dichloromethane (DCM). The organic layer was separated and dried by anhydrous magnesium sulfate. The solvent was removed under reduced pressure and the crude product was purified by flash chromatography (hexane and ethyl acetate 9:1).

O³-Prop-2-ynylestrone (1) ,

White solid, mp 152–153 °C, yield (95%); ¹H NMR (400 MHz, CDCl₃) δ = 7.25 (d, 1H, J 8.4), 6.82–6.80 (dd, 1H, J 8.8, 2.8), 6.74 (d, 1H, J 2.8), 4.69 (d, 2H, J 2.4), 2.94–2.91 (m, 2H), 2.56–2.40 (m, 3H), 2.30–1.96 (m, 5H), 1.67–1.44 (m, 6H), 0.93 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 155.7, 138.0, 133.2, 126.5, 115.1, 112.5, 79.0, 75.5, 55.9, 50.6, 48.2, 44.2, 38.5, 36.0, 31.7, 29.8, 26.7, 26.0, 21.8, 14.0; FT-IR (cm^–1) 3296, 2917, 2866, 2125, 1732, 1497, 1222, 1025, 819, 693; EIMS m/z (%) 308 (M⁺, 100), 280 (4), 251 (8), 223 (12), 184 (13), 149 (48), 85 (24), 71 (29); HRMS (ESI) [m]⁺ calcd for C₂₁H₂₄O₂ 308.1771, found 308.1772.

O³-Prop-2-ynylestradiol (2) ,

White solid, mp 115–116 °C, 0.72 g, yield (78%); ¹H NMR (400 MHz, CDCl₃) δ = 7.25 (d, 1H, J 8.8), 6.81–6.78 (dd, 1H, J 8.8, 2.4), 6.72 (d, 1H, J 2.4), 4.68 (d, 2H, J 2.4), 3.75 (t, 1H, J 8.4), 2.90–2.86 (m, 2H), 2.53 (t, 1H, J 2.4), 2.35–2.13 (m, 3H), 1.99–1.88 (m, 3H), 1.74–1.67 (m, 1H), 1.53–1.21 (m, 6H), 0.80 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 155.4, 138.1, 133.6, 126.3, 114.9, 112.2, 81.9, 78.9, 75.3, 55.7, 50.0, 43.9, 43.2, 38.7, 36.7, 30.6, 29.8, 27.2, 26.2, 23.1, 11.0; FT-IR (cm^–1) 3582, 3288, 2911, 1603, 1496, 1221, 1027, 818, 650; EIMS m/z (%) 310 (M⁺, 100), 272 (5), 251 (8), 210 (6), 197 (13), 171 (17), 144 (31), 115 (17), 91 (12), 77 (6); HRMS (ESI) [m]⁺ calcd for C₂₁H₂₆O₂ 310.1927, found 310.1925

O³-Prop-2-ynyl cholesterol (3) ,

White solid, mp 107–108 °C, 0.87 g, yield (69%); ¹H NMR (400 MHz, CDCl₃) 5.37 (t, 1H, J 5.2), 4.21 (d, 2H, J 2.4), 3.43–3.37 (m, 1H), 2.41 (t, 2H, J 2.4), 2.02–0.87 (m, 39H), 0.69 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 140.8, 122.1, 80.6, 78.4, 73.9, 57.0, 56.3, 55.3, 50.3, 42.5, 40.0, 39.7, 38.9, 37.3, 37.0, 36.4, 36.0, 32.17, 32.10, 28.4, 28.2, 24.5, 24.0, 23.0, 22.7, 21.2, 19.5, 18.9, 12.0; FT-IR (cm^–1) 3402, 2930, 2867, 1712, 1463, 1377, 1054, 1022, 954, 839; EIMS m/z (%) 424 (M⁺, 38), 386 (100), 368 (56), 353 (42), 301 (38), 275 (64), 255 (26), 213 (32), 159 (31), 145 (43), 107 (46), 80 (43); HRMS (ESI) [m]⁺ calcd for C₃₀H₄₈O 424.3700, found 424.3703.

Synthesis of Steroid Esters 4–7

A solution of DCC (5 mmol) and DMAP (0.4 mmol) in 100 mL DCM was added dropwise to a mixture of propiolic acid (5 mmol) and to steroid (4 mmol) in dry dichloromethane (DCM) at 0 °C under nitrogen gas. The reaction mixture was stirred for 3–12 h. The reaction mixture was filtered then washed with 0.1 N HCl (2 × 50 mL) as well as a saturated solution of sodium chloride (2 × 50 mL). The solution was evaporated to give the crude products. The crude products then were purified by silica gel chromatography (90/10 hexane/ethyl acetate).

Estra-1,3,5­(10)-trien-17-one-3-ol-3-(2-propynoate) (4) ,

White solid, mp 208–210 °C, yield (19%); ¹H NMR (400 MHz, CDCl₃) δ = 7.34 (d, 1H, J 8.4), 6.95–6.92 (dd, 1H, J 8.4, 2.4), 6.90 (d, 1H, J 2.4), 3.08 (s, 1H), 2.95–2.92 (dd, 2H, J 8.8, 4.0), 2.57–1.97 (m, 7H), 1.68–1.45 (m, 6H), 0.93 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 151.4, 147.9, 138.5, 138.4, 126.8, 121.4, 118.5, 77.5, 74.5, 50.6, 48.1, 44.3, 38.1, 36.0, 31.7, 29.6, 26.4, 25.9, 21.7, 14.0; FT-IR (cm^–1) 3207, 2929, 2862, 2110, 1725, 1652, 1453, 1190, 1010, 906, 802; EIMS m/z (%) 322 (M⁺, 100), 265 (26), 252 (14), 172 (12), 158 (21), 115 (19), 91 (13); HRMS (ESI) [m]⁺ calcd for C₂₁H₂₂O₃ 322.1563, found 322.1561.

5-Androst-en-17-one-3-ol-3-(2-propynoate) (5) ,

White solid, mp 128–130 °C, yield (46%); ¹H NMR (400 MHz, CDCl₃) δ = 5.45–5.44 (dd, 1H, J 4.0, 0.8), 4.76–4.71 (m, 1H), 2.89 (s, 1H), 2.51–2.41 (m, 3H), 2.15–2.06 (m, 2H), 1.98–1.84 (m, 6H), 1.72–1.48 (m, 6H), 1.34–1.16 (m, 3H), 1.07–1.06 (m, 4H), 0.90 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 152.3, 139.5, 122.6, 76.3, 75.2, 74.4, 51.8, 50.3, 47.7, 37.9, 37.0, 36.9, 36.0, 31.6, 31.5, 30.9, 27.6, 22.0, 20.5, 19.4, 13.7; FT-IR (cm^–1) 3215, 2954, 2906, 2105, 1729, 1705, 1226, 1018, 977, 761; EIMS m/z (%) 340 (M⁺, 16), 270 (100), 213 (9), 145 (17), 121 (48), 91 (37); HRMS (ESI) [m]⁺ calcd for C₂₂H₂₈O₃ 340.2033, found 340.2033.

Pregn-5-en-20-one-3-ol-3-(2-propynoate) (6)

White solid, mp 205–207 °C, yield (43%); ¹H NMR (600 MHz, CDCl₃) δ = 5.42 (t, 1H, J 1.6), 4.78–4.72 (m, 1H), 2.89 (s, 1H), 2.55 (t, 1H, J 9.0), 2.42 (t, 1H, J 4.2), 2.21–2.19 (m, 1H), 2.14 (s, 3H), 2.08–1.91 (m, 4H), 1.72–1.47 (m, 9H), 1.24–1.02 (m, 7H), 0.65 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 209.7, 152.3, 139.3, 123.1, 75.2, 74.4, 63.8, 57.0, 50.0, 44.1, 38.9, 37.9, 37.0, 36.7, 31.99, 31.96, 31.7, 27.6, 24.6, 23.0, 21.2, 19.4, 13.4; FT-IR (cm^–1)­3303, 3203, 2934, 2853, 2107, 1725, 1700, 1621, 1593, 1388, 1237, 1081, 769, 741; EIMS m/z (%) 368 (M⁺, 5), 298 (100), 283 (18), 255 (7), 213 (9), 195 (57), 151 (57), 113 (63), 98 (47), 70 (66); HRMS (ESI) [m]⁺ calcd for C₂₄H₃₂O₃ 368.2346, found 368.2346.

Testosterone-17-(2-propynoate) (7) ,

White solid, mp 198–200 °C, yield (12%); ¹H NMR (600 MHz, CDCl₃) δ = 5.75 (s, 1H), 4.72 (t, 1H, J 7.8), 2.90 (s, 1H), 2.44–2.22 (m, 5H), 2.06–2.02 (m, 1H), 1.87–1.84 (m, 2H), 1.74–1.69 (m, 2H), 1.64–1.59 (m, 3H), 1.44–1.39 (m, 2H), 1.24–1.20 (m, 4H), 1.11–0.95 (m, 3H), 0.90 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 199.6, 170.9, 152.9, 124.1, 84.8, 75.1, 74.6, 53.8, 50.3, 42.9, 38.7, 36.7, 35.8, 35.5, 34.1, 32.8, 31.6, 27.4, 23.6, 20.6, 17.5, 12.2; FT-IR (cm^–1) 3173, 2946, 2103, 1704, 1659, 1440, 1227, 1004, 865, 750; EIMS m/z (%) 340 (M⁺, 32), 298 (69), 255 (41), 228 (36), 185 (19), 147 (59), 124 (100), 91 (31); HRMS (ESI) [m]⁺ calcd for C₂₂H₂₈O₃ 340.2033, found 340.2034.

Synthesis of Triazoles 8–56

To a solution of amines in hydrochloric acid, sodium nitrite solution was added dropwise at 0–5 °C then stirred for 1 h to afford the diazonium chloride salt. This was followed by the addition of a sodium azide to diazonium salt solution to afford aryl azides.

Click reaction involved stirring of steroid ethers and esters 1–7 with aryl azides in the presence of catalytic amount of copper sulfate (5%) and sodium ascorbate (2%) in 50 mL ter. butanol/water (7:3) as the solvent. The heterogeneous mixture was stirred under reflux overnight. TLC analysis was then used to indicate complete consumption of the reactants. The crude product was purified by flash chromatography. The reaction gave the desired triazoles 8–56 in good to excellent yields (62–95%).

13-Methyl-3-((1-phenyl-1H-1,2,3-triazol-4-yl)­methoxy)-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (8)

White solid, mp 194–196 °C, yield (87%); ¹H NMR (400 MHz, DMSO) δ = 8.93 (s, 1H), 7.93 (d, 2H, J 7.6), 7.61 (t, 2H, J 7.2), 7.50 (t, 1H, J 6.8), 7.22 (d, 1H, J 8.8), 6.84 (d, 1H, J 8.8), 6.79 (d, 1H, J 2.4), 5.19 (s, 2H), 2.85–2.83 (m, 2H), 2.43–2.36 (m, 2H), 2.20–1.93 (m, 6H), 1.77–1.75 (m, 1H), 1.59–1.37 (m, 4H), 0.83 (s, 3H); ¹³C­{¹H} NMR (100 MHz, DMSO) δ = 155.8, 144.1, 137.5, 136.5, 132.1, 129.8, 128.7, 126.2, 122.6, 120.0, 114.4, 112.2, 60.8, 49.5, 47.2, 43.4, 37.7, 35.3, 30.6, 29.1, 26.0, 25.4, 21.1, 13.4; FT-IR (cm^–1) 3139, 2932, 2862, 1741, 1602, 1500, 1229, 1050, 1009, 819, 760, 687; EIMS m/z (%) 427 (M⁺, 9), 398 (6), 371 (8), 270 (26), 235 (6), 185 (4), 130 (100), 77 (19); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₉O₂N₃ 427.2254, found 427.2253.

3-((1-(4-Chlorophenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (9)

White solid, mp 208–210 °C, yield (88%); ¹H NMR (400 MHz, CDCl₃) δ = 8.05 (s, 1H), 7.73–7.70 (dd, 2H, J 6.8, 2.4), 7.53–7.51 (dd, 2H, J 6.8, 2.4), 7.25 (d, 1H, J 8.4), 6.85–6.83 (dd, 1H, J 8.4, 2.4), 6.79 (d, 1H, J 2.4), 5.29 (s, 2H), 2.93–2.91 (m, 2H), 2.65–1.96 (m, 8H), 1.70–1.37 (m, 5H), 0.93 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.1, 138.0, 135.4, 134.7, 132.9, 129.9, 126.5, 121.7, 120.7, 114.7, 112.3, 61.9, 50.4, 8.0, 44.0, 38.3, 35.8, 31.5, 29.6, 26.5, 25.9, 21.6, 13.8; FT-IR (cm^–1) 3135, 2940, 1744, 1605, 1501, 1462, 1228, 1032, 1011, 827, 720, 513; EIMS m/z (%) 461 (M⁺, 14), 405 (22), 282 (9), 270 (91), 242 (4), 185 (8), 164 (100), 128 (19), 111 (26), 75 (8); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₈O₂N₃Cl 461.1865, found 461.1866).

3-((1-(4-Bromophenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (10)

White solid, mp 204–206 °C, yield (92%); ¹H NMR (400 MHz, CDCl₃) δ = 8.05 (s, 1H), 7.69–7.64 (m, 4H), 7.25 (d, 1H, J 8.4), 6.86–6.83 (dd, 1H, J 8.4, 2.4), 6.78 (d, 1H, J 2.8), 5.29 (s, 2H), 2.92–2.91 (m, 2H), 2.65–1.96 (m, 8H), 1.67–1.27 (m, 5H), 0.93 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 165.3, 138.2,136.1, 133.16, 13.10, 126.7, 122.7, 122.1, 120.8, 114.9, 112.5, 62.1, 50.6, 48.2, 44.2, 38.5, 36.0, 31.7, 29.8, 26.7, 26.1, 21.8, 14.0; FT-IR (cm^–1) 3134, 2920, 2887, 1744, 1605, 1499, 1229, 1033, 823, 511; EIMS m/z (%) 505 (M⁺, 11), 449 (18), 312 (9), 270 (100), 208 (50), 155 (22), 129 (100), 102 (11); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₈O₂N₂Br 505.1359, found 505.1364.

3-((1-(4-Iodophenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (11)

White solid, mp 220–222 °C, yield (81%); ¹H NMR (600 MHz, DMSO) δ = 8.94 (s, 1H), 7.97–7.94 (dt, 2H, J 9.6, 3.0), 7.75–7.72 (dt, 2H, J 9.6, 2.4), 7.19 (d, 1H, J 9.0), 6.82–6.80 (dd, 1H, J 8.4, 2.4), 6.77 (d, 1H, J 2.4), 5.17 (s, 2H), 2.84–2.82 (m, 2H), 2.45–1.91 (m, 6H), 1.76–1.33 (m, 7H), 0.82 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.3, 139.3, 138.2, 133.1, 126.7, 126.6, 122.6, 115.0, 112.6, 93.8, 61.8, 50.6, 48.2, 44.1, 38.5, 36.0, 31.7, 29.9, 26.7, 26.1, 21.7, 14.0; FT-IR (cm^–1) 3091, 2933, 2864, 1732, 1607, 1494, 377, 1245, 1025, 820, 514; EIMS m/z (%) 553 (M⁺, 4), 497 (6), 361 (6), 280 (7), 270 (55), 256 (38), 213 (6), 185 (12), 129 (100), 102 (19), 76 (34); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₈O₂N₃I 553.1221, found 553.1223.

3-((1-(4-Azidophenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (12)

White solid, mp 201–203 °C, yield (89%); ¹H NMR (600 MHz, DMSO) δ = 8.95 (s, 1H), 7.94–7–7.94 (dt, 2H, J 10.2, 3.0), 7.69–7.66 (dt, 2H, J 10.2, 3.0), 7.19 (d, 1H, J 9.0), 6.83–6.81 (dd, 1H, J 8.4, 2.4), 6.78 (d, 1H, J 3.0), 5.17 (s, 2H), 2.84–2.82 (m, 2H), 2.45–1.92 (m, 6H), 1.76–1.33 (m, 7H), 0.82 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.8, 144.3, 137.5, 135.3, 132.9, 132.2, 129.8, 126.2, 122.7, 121.7, 114.4, 112.2, 60.8, 49.5, 47.2, 43.4, 37.7, 35.3, 31.3, 29.1, 26.0, 25.4, 21.1, 13.4; FT-IR (cm^–1) 3136, 2942, 2881, 1744, 1606, 1501, 1310, 1229, 1096, 1033, 1011, 828, 514; EIMS m/z (%) 468 (M⁺, 10), 405 (7), 270 (52), 213 (7), 185 (10), 164 (100), 111 (26); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₈O₂N₆ 468.2268, found 468.2265.

13-Methyl-3-((1-(4-nitrophenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (13)

Yellow solid, mp 220–222 °C, yield (83%); ¹H NMR (600 MHz, DMSO) δ = 9.14 (s, 1H), 8.45 (d, 2H, J 8.4), 8.24 (d, 2H, J 8.4), 7.19 (d, 1H, J 8.4), 6.82 (d, 1H, J 8.4), 6.78 (s, 1H), 5.20 (s, 2H), 2.84–2.82 (m, 2H), 2.45–1.93 (m, 6H), 1.75–1.33 (m, 7H), 0.82 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.7, 146.7, 144.8, 140.7, 137.5, 132.2, 126.2, 125.5, 123.1, 120.6, 116.6, 114.4, 114.1, 112.2, 60.7, 49.5, 47.2, 43.4, 37.7, 35.3, 31.3, 31.2, 29.1, 25.9, 25.4, 21.1, 13.4; FT-IR (cm^–1) 3130, 2931, 2878, 1732, 1696, 1527, 1341, 1235, 1039, 850, 748, 683; EIMS m/z (%) 472 (M⁺, 12), 443 (10), 416 (9), 292 (6), 280 (10), 270 (100), 251 (4), 213 (12), 185 (18), 175 (46), 145 (39), 129 (42), 91 (21), 76 (11); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₈O₄N₄ 472.2105, found 472.2103.

3-((1-(4-Acetylphenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (14)

Yellow solid, mp 185–187 °C, yield (82%); ¹H NMR (600 MHz, DMSO) δ = 9.08 (s, 1H), 8.18–8.17 (dt, 2H, J 9.0, 1.8), 8.12–8.10 (dt, 2H, J 9.0, 2.4), 7.21 (d, 1H, J 8.4), 6.85–6.83 (dd, 1H, J 8.4, 2.4), 6.80 (d, 1H, J 3.0), 5.21 (s, 2H), 2.86–2.83 (m, 2H), 2.65 (s, 3H), 2.46–1.93 (m, 6H), 1.77–1.36 (m, 7H), 0.83 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.8, 144.3, 138.5, 137.5, 136.2, 132.2, 126.2, 122.6, 121.9, 114.4, 112.2, 94.4, 60.8, 49.5, 47.3, 43.4, 40.1, 37.7, 35.3, 31.3, 29.1, 26.0, 25.4, 21.1, 13.4; FT-IR (cm^–1) 3146, 2930, 2867, 1734, 1685, 1603, 1496, 1358, 1248, 1029, 842, 590; EIMS m/z (%) 469 (M⁺, 12), 440 (8), 413 (14), 289 (8), 270 (100), 213 (9), 185 (12), 172 (88), 130 (42), 77 (6); HRMS (ESI) [m]⁺ calcd for C₂₉H₃₁O₃N₃ 469.23, found 469.2361.

4-(4-(((13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1h-1,2,3-triazol-1-yl)­benzenesulfonamide (15)

White solid, mp 220–222 °C, yield (86%); ¹H NMR (600 MHz, DMSO) δ = 9.04 (s, 1H), 8.16 (d, 2H, J 12), 8.04 (d, 2H, J 12), 7.54 (s, 2H), 7.20 (d, 1H, J 6.0), 6.84 (d, 1H, J 12), 6.79 (s, 1H), 5.20 (s, 2H), 2.84–2.83 (m, 2H), 2.45–2.34 (m, 2H), 2.18–1.93 (m, 4H), 1.76–1.33 (m, 7H), 0.83 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.8, 144.5, 143.8, 138.5, 137.5, 132.2, 127.4, 126.2, 122.9, 120.3, 114.4, 112.2, 60.8, 49.5, 47.2, 43.4, 37.7, 35.3, 31.3, 29.1, 26.0, 25.4, 21.1, 13.4; FT-IR (cm^–1) 3226, 2927, 2864, 1732, 1595, 1499, 1345, 1230, 1162, 1054, 824, 612, 549; EIMS m/z (%) 506 (M⁺, 8), 424 (6), 308 (8), 270 (100), 226 (6), 213 (18), 185 (28), 172 (32), 146 (24), 130 (56), 115 (12), 77 (16); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₀O₄N₄S 506.1982, found 506.1983.

13-Methyl-3-((1-(4-(trifluoromethyl)­phenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (16)

White solid, mp 188–190 °C, yield (65%); ¹H NMR (600 MHz, DMSO) δ = 9.09 (s, 1H), 8.21 (d, 2H, J 8.4), 8.01 (d, 2H, J 10.2, 3.0), 7.19 (d, 1H, J 9.0), 6.83–6.81 (dd, 1H, J 9.0), 7.21 (d, 1H, J 8.4), 6.85–6.83 (dd, 1H, J 8.4, 2.4), 6.80 (d, 1H, J 2.4), 5.21 (s, 2H), 2.85–2.83 (m, 2H), 2.46–1.93 (m, 6H), 1.77–1.34 (m, 7H), 0.83 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.8, 144.5, 139.3, 137.5, 132.2, 127.23, 127.21, 126.2, 122.9, 120.5, 114.5, 112.3, 60.8, 49.5, 47.2, 43.4, 37.7, 35.3, 31.2, 29.1, 26.0, 25.4, 21.1, 13.4; FT-IR (cm^–1) 2929, 2867, 1736, 1609, 1497, 1321, 1249, 1165, 1116, 1066, 1029, 846, 595; EIMS m/z (%) 495 (M⁺, 14), 466 (8), 439 (12), 316 (9), 303 (14), 270 (100), 242 (4), 213 (10), 198 (94), 185 (16), 145 (42), 129 (8), 91 (5); HRMS (ESI) [m]⁺ calcd for C₂₈H₂₈O₂N₂F₃ 495.2128, found 495.2128.

2-(4-(((13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1h-1,2,3-triazol-1-yl)­benzoic Acid (17)

White solid, mp 190–192 °C, yield (83%); ¹H NMR (600 MHz, DMSO) δ = 8.59 (s, 1H), 7.92 (d, 1H, J 7.2), 7.74 (t, 1H, J 7.8), 7.68 (t, 1H, J 7.6), 7.61 (d, 1H, J 7.8), 7.20 (d, 1H, J 8.4), 6.83–6.81 (dd, 1H, J 8.4, 2.4), 6.78 (d, 1H, J 2.4), 5.14 (s, 2H), 2.84–2.81 (m, 2H), 2.45–1.92 (m, 6H), 1.76–1.34 (m, 7H), 0.82 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 156.1, 143.0, 137.6, 135.2, 132.4, 132.3, 130.0, 126.39, 126.34, 126.0, 114.5, 112.46, 112.43, 78.0, 61.0, 49.6, 47.4, 43.5, 37.9, 37.8, 35.4, 31.4, 29.2, 26.1, 26.0, 25.59, 25.56, 21.2, 13.6; FT-IR (cm^–1) 2926, 2879, 1729, 1720, 1604, 1500, 1240, 1057, 800, 762, 703, 638; EIMS m/z (%) 471 (M⁺, 8), 424 (6), 370 (8), 307 (4), 270 (52), 213 (10), 185 (14), 146 (16), 130 (100), 103 (8), 77 (25); HRMS (ESI) [m]⁺ calcd for C₂₈H₂₉O₄N₃ 471.2153, found 471.2158.

13-Methyl-3-((1-(naphthalen-1-Yl)-1h-1,2,3-triazol-4-yl)­methoxy)-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (18)

White solid, mp 170–172 °C, yield (90%); ¹H NMR (400 MHz, CDCl₃) δ = 8.05–8.00 (m, 1H), 8.00 (s, 1H), 7.99 (d, 1H, J 7.2), 7.65–7.55 (m, 5H), 7.27 (d, 1H, J 8.4), 6.90–6.88 (dd, 1H, J 8.8, 2.8), 6.83 (d, 1H, J 2.8), 5.38 (s, 2H), 2.95–2.92 (m, 2H), 2.56–2.40 (m, 2H), 2.29–1.97 (m, 5H), 1.65–1.44 (m, 6H), 0.93 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.5, 138.2, 134.3, 133.8, 133.0, 130.7, 128.6, 128.5, 128.1, 127.3, 126.7, 125.6, 125.1, 123.8, 122.5, 115.1, 112.6, 62.3, 50.6, 48.2, 44.2, 38.5, 36.0, 31.8, 29.8, 26.7, 26.1, 21.8, 14.0; FT-IR (cm^–1) 3143, 2925, 2861, 2827, 1725, 1609, 1572, 1498, 1371, 1248, 1158, 1008, 805, 772; EIMS m/z (%) 477 (M⁺, 5), 421 (8), 367 (6), 313 (6), 270 (10), 239 (4), 180 (100), 152 (8), 127 (18), 98 (6), 71 (11), 57 (21); HRMS (ESI) [m]⁺ calcd for C₃₁H₃₁O₂N₃ 477.2411, found 477.2409.

3-((1-Benzyl-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (19)

White solid, mp 185–187 °C, yield (78%); ¹H NMR (400 MHz, CDCl₃) δ = 7.55 (s, 1H), 7.41–7.38 (m, 3H), 7.31–7.29 (m, 2H), 7.22 (d, 1H, J 8.8), 6.80–6.77 (dd, 1H, J 8.4, 2.8), 6.73 (d, 1H, J 2.8), 5.55 (s, 2H), 5.19 (s, 2H), 2.92–2.88 (m, 2H), 2.55–2.39 (m, 2H), 2.26–1.95 (m, 5H), 1.64–1.45 (m, 6H), 0.92 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.3, 144.9, 138.1, 132.9, 129.4, 129.1, 128.3, 126.6, 122.8, 114.9, 112.6, 62.1, 54.6, 50.6, 48.2, 44.1, 38.5, 36.0, 31.7, 29.8, 26.7, 26.0, 21.7, 14.0; FT-IR (cm^–1) 2922, 1732, 1611, 1497, 1452, 1279, 1246, 1218, 1064, 1047, 903, 817, 730, 579; EIMS m/z (%) 441 (M⁺, 28), 412 (4), 384 (5), 270 (19), 213 (4), 172 (6), 144 (100), 115 (8), 91 (97), 65 (5); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₁O₂N₃ 441.2411, found 441.2411.

3-((1-(3,4-Dimethylphenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (20)

Yellow solid, mp 189–191 °C, yield (75%); ¹H NMR (600 MHz, DMSO) δ = 8.84 (s, 1H), 7.71 (d, 1H, J 1.8), 7.61–7.59 (dd, 1H, J 7.8, 1.8), 7.34 (d, 1H, J 8.4), 7.19 (d, 1H, J 8.4), 6.83–6.81 (dd, 1H, J 8.4, 3.0), 6.77 (d, 1H, J 2.4), 5.16 (s, 2H), 2.45–2.33 (m, 2H), 2.31 (s, 3H), 2.28 (s, 3H), 2.19–1.74 (m, 5H), 1.56–1.34 (m, 6H), 0.82 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.8, 143.9, 138.1, 137.5, 137.0, 134.4, 132.1, 130.5, 126.2, 122.4, 120.9, 119.3, 117.3, 116.5, 116.0, 114.5, 112.9, 112.2, 110.9, 60.9, 49.5, 47.3, 43.4, 37.7, 35.3, 31.3, 29.1, 26.0, 25.4, 21.1, 19.4, 18.9, 13.4; FT-IR (cm^–1) 2951, 2923, 2856, 1735, 1613, 1577, 1500, 1465, 1280, 1225, 1062, 1040, 877, 824, 702; EIMS m/z (%) 455 (M⁺, 6), 399 (10), 270 (8), 158 (100), 105 (8), 77 (8); HRMS (ESI) [m]⁺ calcd for C₂₉H₃₃O₂N₃ 455.2567, found 455.2568.

3-((1-Mesityl-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (21)

White solid, mp 200–202 °C, yield (81%); ¹H NMR (400 MHz, CDCl₃) δ = 7.64 (s, 1H), 7.24 (d, 1H, J 8.4), 7.00 (s, 2H), 6.85–6.83 (dd, 1H, J 8.4, 2.8), 6.78 (d, 1H, J 2.8), 5.31 (s, 2H), 2.91–2.89 (m, 2H), 2.56–2.54 (m, 1H), 2.40 (s, 3H), 2.37–2.14 (m, 5H), 1.97 (s, 6H), 1.64–1.49 (m, 7H), 0.93 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.5, 144.5, 140.2, 138.0, 135.2, 132.9, 129.2, 126.5, 124.6, 115.2, 112.8, 62.6, 50.6, 48.1, 44.1, 38.5, 36.0, 31.7, 29.8, 26.7, 26.0, 21.7, 21.3, 17.4, 14.0; FT-IR (cm^–1) 2925, 2862, 1733, 1608, 1495, 1375, 1280, 1240, 1040, 851, 579; EIMS m/z (%) 469 (M⁺, 8), 398 (2), 270 (2), 172 (100), 157 (10), 119 (4), 91 (4); HRMS (ESI) [m]⁺ calcd for C₃₀H₃₅O₂N₃ 469.2724, found 469.2724.

3-((1-(4-Iodo-2-methylphenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (22)

White solid, mp 207–209 °C, yield (73%); ¹H NMR (400 MHz, CDCl₃) δ = 7.79 (s, 1H), 7.77 (d, 1H, J 1.2), 7.71–7.68 (ddd, 1H, J 8.4, 2.0, 0.4), 7.25 (d, 1H, J 8.4), 7.10 (d, 1H, J 8.4), 6.86–6.83 (dd, 1H, J 8.8, 2.8), 6.79 (d, 1H, J 2.4), 5.30 (s, 2H), 2.94–2.91 (m, 2H), 2.56–2.40 (m, 2H), 2.30–1.96 (m, 7H), 1.71–1.46 (m, 7H), 0.93 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.4, 144.8, 140.6, 138.1, 136.3, 136.2, 136.0, 133.0, 127.6, 126.6, 124.2, 155.0, 112.6, 95.8, 62.3, 50.6, 48.2, 44.2, 38.5, 36.0, 31.7, 29.8, 26.7, 26.1, 21.8, 17.8, 14.0; FT-IR (cm^–1) 3087, 2978, 2925, 1734, 1607, 1492, 1376, 1242, 1028, 817, 558; EIMS m/z (%) 567 (M⁺, 10), 510 (9), 374 (12), 307 (3), 270 (100), 213 (4), 178 (17), 143 (100), 90 (16); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₀O₂N₃I 567.1377, found 567.1378.

3-((1-(3,5-Bis­(trifluoromethyl)­phenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-6,7,8,9,11,12,13,14,15,16-decahydro-17H-cyclopenta­[a]­phenanthren-17-one (23)

White solid, mp 190–192 °C, yield (62%); ¹H NMR (600 MHz, DMSO) δ = 9.26 (s, 1H), 8.66 (s, 2H), 8.26 (s, 1H), 7.20 (d, 1H, J 8.4), 6.83–6.81 (dd, 1H, J 8.4, 1.8), 6.78 (s, 1H), 5.22 (s, 2H), 2.83–2.82 (m, 2H), 2.45–1.93 (m, 6H), 1.76–1.33 (m, 7H), 0.82 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.8, 144.7, 137.8, 137.5, 132.3, 132.0, 131.6, 126.2, 124.1, 123.3, 122.0, 121.4, 120.87, 120.84, 114.5, 112.2, 60.9, 49.5, 47.3, 43.4, 37.7, 35.5, 31.3, 29.1, 26.0, 25.4, 21.1, 13.4; FT-IR (cm^–1) 3063, 2931, 2864, 1729, 1607, 1495, 1356, 1277, 1137, 1036, 895, 815, 681; EIMS m/z (%) 563 (M⁺, 10), 497 (11), 382 (8), 369 (12), 341 (7), 270 (100), 246 (12), 213 (48), 185 (21), 146 (16), 133 (13), 69 (8); HRMS (ESI) [m]⁺ calcd for C₂₉H₂₇O₂N₃F₆ 563.2002, found 563.2002.

2-(4-(((13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)-N-(3-nitrophenyl)­benzamide (24)

Pale yellow solid, mp 250–252 °C, yield (91%); ¹H NMR (600 MHz, DMSO) δ = 10.99 (s, 1H), 8.63 (s, 1H), 8.59 (t, 1H, J 1.8), 7.95–7.94 (dd, 1H, J 7.8, 1.2), 7.87 (d, 1H, J 7.8), 7.83 (d, 1H, J 7.2), 7.77–7.69 (m, 3H), 7.60 (t, 1H, J 8.4), 7.15 (d, 1H, J 8.4), 6.77–6.76 (dd, 1H, J 8.4, 2.4), 6.72 (d, 1H, J 1.8), 5.12 (s, 2H), 2.79–2.78 (m, 2H), 2.44–1.90 (m, 6H), 1.76–1.34 (m, 7H), 0.81 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 165.4, 156.0, 147.9, 143.4, 140.0, 137.6, 134.0, 132.2, 132.1, 131.6, 130.3, 129.7, 129.2, 126.3, 125.7, 125.4, 125.2, 118.4, 114.5, 113.8, 112.3, 67.1, 60.9, 49.6, 47.4, 43.5, 37.9, 35.4, 31.4, 29.2, 26.1, 25.5, 25.2, 21.2, 13.5; FT-IR (cm^–1) 2927, 2864, 1732, 1681, 1604, 1528, 1497, 1429, 1350, 1246, 1161, 819, 737, 672; EIMS m/z (%) 591 (M⁺, 10), 496 (4), 470 (8), 423 (8), 293 (24), 270 (100), 213 (22), 185 (34), 146 (36), 120 (32), 92 (16), 76 (24); HRMS (ESI) [m]⁺ calcd for C₃₄H₃₃O₅N₅ 591.2476, found 591.2475.

2-(4-(((13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)-N-(4-(trifluoromethyl)­phenyl)­benzamide (25)

Pale yellow solid, mp 230–233 °C, yield (94%); ¹H NMR (600 MHz, DMSO) δ = 10.84 (s, 1H), 8.61 (s, 1H), 7.80 (d, 1H, J 7.8), 7.63–7.68 (m, 5H), 7.66 (d, 2H, J 7.8), 7.16 (d, 1H, J 8.4), 6.78–6.76 (dd, 1H, J 8.4, 1.8), 6.73 (s, 1H), 5.12 (s, 2H), 2.80–2.79 (m, 2H), 2.45–1.91 (m, 6H), 1.75–1.32 (m, 7H), 0.81 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 165.3, 156.0, 143.4, 142.4, 137.6, 134.0, 132.3, 132.2, 131.4, 129.7, 129.2, 126.3, 126.1, 126.0, 125.5, 125.3, 125.2, 119.7, 114.6, 112.4, 67.1, 60.9, 49.6, 47.4, 43.5, 37.9, 35.4, 31.4, 29.2, 26.1, 25.5, 21.2, 13.5; FT-IR (cm^–1) 3272, 2928, 2860, 1734, 1605, 1532, 1498, 1408, 1320, 1252, 1161, 113, 1064, 840, 761; EIMS m/z (%) 614 (M⁺, 12), 558 (8), 425 (14), 356 (17), 315 (32), 299 (72), 271 (65), 213 (38), 172 (45), 156 (100), 146 (68), 77 (28); HRMS (ESI) [m]⁺ calcd for C₃₅H₃₃O₃N₄F₃ 614.2499, found 614.2500.

2-(4-(((13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)-N-(naphthalen-1-yl)­benzamide (26)

Yellow solid, mp 215–217 °C, yield (78%); ¹H NMR (600 MHz, DMSO) δ = 10.52 (s, 1H), 8.63 (s, 1H), 7.96–7.92 (m, 3H), 7.83 (d, 1H, J 8.4), 7.77–7.73 (m, 3H), 7.58–7.49 (m, 4H), 7.15 (d, 1H, J 9.0), 6.81–6.79 (dd, 1H, J 8.4, 2.4), 6.75 (d, 1H, J 2.4), 5.16 (s, 2H), 2.79–2.78 (m, 2H), 2.44–2.03 (m, 6H), 1.77–1.31 (m, 7H), 0.81 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 165.9, 156.0, 143.4, 137.6, 134.1, 133.7, 133.1, 132.2, 131.0, 129.7, 129.2, 128.6, 128.0, 126.3, 126.2, 126.1, 126.0, 125.59, 125.53, 125.2, 123.2, 122.9, 114.5, 112.3, 60.9, 54.9, 49.6, 47.4, 43.5, 37.9, 35.4, 31.4, 29.2, 26.1, 25.5, 21.2, 13.5; FT-IR (cm^–1) 3301, 3155, 3056, 2936, 2825, 1737, 1679, 1538, 1498, 1344, 1250, 1027, 771; EIMS m/z (%) 596 (M⁺, 11), 528 (24), 423 (18), 322 (6), 299 (100), 242 (9), 213 (21), 185 (33), 156 (61), 127 (58), 77 (19); HRMS (ESI) [m]⁺ calcd for C₃₈H₃₆O₃N₄ 596.2782, found 596.2780.

13-Methyl-3-((1-phenyl-1h-1,2,3-triazol-4-yl)­methoxy)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (27)

White solid, mp 145–147 °C, yield (83%); ¹H NMR (400 MHz, CDCl₃) δ = 8.33 (s, 1H), 7.80 (d, 2H, J 7.2), 7.56 (t, 2H, J 7.2), 7.48 (t, 1H, J 7.4), 7.26 (d, 1H, J 7.4), 6.87 (d, 1H, J 7.2), 6.80 (s, 1H), 5.28 (s, 2H), 3.76 (t, 1H, J 7.8), 2.90–1.89 (m, 5H), 1.75–1.19 (m, 11H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.2, 138.3, 138.2, 133.6, 130.1, 129.0, 126.6, 121.0, 115.0, 114.9, 112.4, 82.0, 55.9, 50.2, 44.1, 43.4, 38.9, 36.8, 31.4, 30.7, 30.0, 27.3, 26.4, 23.3, 11.2; FT-IR (cm^–1) 3306, 2923, 2867, 1601, 1499, 1465, 1231, 1040, 1010, 758, 688; EIMS m/z (%) 429 (M⁺, 11), 400 (8), 310 (7), 272 (41), 235 (9), 159 (8), 130 (100), 103 (7), 77 (32); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₁O₂N₃ 429.2411, found 429.2414.

3-((1-(4-Chlorophenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (28)

White solid, mp 204–206 °C, yield (87%); ¹H NMR (400 MHz, CDCl₃) δ = 8.06 (s, 1H), 7.73–7.70 (dt, 2H, J 9.2, 2.4), 7.54–7.51 (dt, 2H, J 9.2, 2.4), 7.26 (d, 1H, J 8.4), 6.85–6.82 (dd, 1H, J 8.4, 2.4), 6.77 (d, 1H, J 2.4), 5.29 (s, 2H), 3.75 (t, 1H, J 8.8), 2.88–2.86 (m, 2H), 2.32–2.14 (m, 3H), 2.13–1.53 (m, 4H), 1.52–1.21 (m, 7H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.2, 138.4, 134.8, 133.7, 130.1, 126.6, 121.9, 114.9, 112.4, 82.1, 62.2, 50.2, 44.1, 43.4, 39.0, 36.9, 30.8, 30.0, 27.4, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3409, 3139, 2926, 2870, 1608, 1502, 1229, 1031, 1010, 328, 512; EIMS m/z (%) 463 (M⁺, 16), 407 (19), 309 (4), 282 (6), 272 (100), 240 (4), 213 (16), 164 (78), 128 (17), 111 (22), 75 (8); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₀O₂N₃Cl 463.2021, found 463.2022.

3-((1-(4-Bromophenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (29)

White solid, mp 198–200 °C, yield (90%); ¹H NMR (400 MHz, CDCl₃) δ = 8.07 (s, 1H), 7.70–7.64 (m, 4H), 7.26 (d, 1H, J 8.8), 6.85–6.82 (dd, 1H, J 8.8, 2.4), 6.77 (d, 1H, J 2.4), 5.29 (s, 2H), 3.75 (t, 1H, J 8.8), 2.90–2.86 (m, 2H), 2.35–2.12 (m, 3H), 1.99–1.52 (m, 3H), 1.51–1.22 (m, 8H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.2, 138.4, 133.7, 133.1, 126.7, 122.7, 122.1, 114.9, 112.4, 82.1, 62.2, 50.2, 44.1, 43.4, 39.0, 36.9, 30.8, 30.0, 27.4, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3393, 3138, 3106, 2921, 2869, 1608, 1498, 1250, 1228, 1030, 1009, 823, 699; EIMS m/z (%) 507 (M⁺, 8), 451 (11), 315 (8), 272 (100), 254 (3), 208 (31), 172 (7), 157 (14), 129 (73), 102 (8); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₀O₂N₃Br 507.1516, found 507.1517.

3-((1-(4-Iodophenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (30)

White solid, mp 216–218 °C, yield (90%); ¹H NMR (400 MHz, CDCl₃) δ = 8.36 (s, 1H), 7.89 (d, 2H, J 7.8), 7.56 (d, 2H, J 7.8), 7.26 (d, 1H, J 7.2), 6.86 (d, 1H, J 6.5), 6.78 (s, 1H), 5.27 (s, 2H), 3.75 (t, 1H, J 8.4), 2.88–2.84 (m, 2H), 2.35–2.10 (m, 3H), 1.99–1.89 (m, 2H), 1.76–1.18 (m, 9H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.1, 139.2, 138.4, 133.7, 126.6, 122.4, 114.9, 112.4, 93.8, 82.1, 61.9, 50.2, 44.1, 43.4, 39.0, 36.9, 30.8, 30.0, 27.4, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3371, 3129, 2919, 2869, 1607, 1496, 1310, 1249, 1231, 1052, 2032, 1011, 816, 509; EIMS m/z (%) 555 (M⁺, 14), 499 (18), 399 (42), 330 (13), 310 (26), 281 (100), 208 (30), 150 (46), 71 (34); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₀O₂N₃I 555.1377, found 555.1377.

3-((1-(4-Azidophenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (31)

White solid, mp 198–200 °C, yield (85%); ¹H NMR (400 MHz, CDCl₃) δ = 8.08 (s, 1H), 7.73 (d, 2H, J 8.8), 7.54–7.51 (dt, 2H, J 8.8, 2.8), 7.26 (d, 1H, J 8.4), 6.84 (d, 1H, J 8.4), 6.77 (s, 1H), 5.29 (s, 2H), 3.75 (t, 1H, J 8.4), 2.88–2.86 (m, 2H), 2.31–1.58 (m, 6H), 1.54–1.21 (m, 8H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.0, 138.2, 133.5, 129.9, 126.5, 121.7, 114.7, 112.2, 81.9, 62.0, 50.0, 43.9, 43.2, 38.8, 36.7, 30.6, 29.8, 27.2, 26.3, 23.1, 11.0; FT-IR (cm^–1) 3292, 2923, 2868, 1684, 1605, 1498, 1229, 1050, 1024, 828, 632, 513; EIMS m/z (%) 470 (M⁺, 4), 463 (6), 434 (10), 407 (9), 282 (4), 272 (92), 240 (4), 213 (12), 186 (5), 164 (100), 146 (10), 128 (14), 111 (22), 75 (8); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₀O₂N₆ 470.2425, found 470.2424.

13-Methyl-3-((1-(4-nitrophenyl)-1h-1,2,3-triazol-4-yl)­methoxy)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (32)

Yellow solid, mp 215–217 °C, yield (82%); ¹H NMR (400 MHz, CDCl₃) δ = 8.46 (d, 2H, J 8.8), 8.29 (s, 1H), 8.03 (d, 2H, J 8.8), 7.27 (d, 1H, J 8.0), 6.85 (d, 1H, J 8.4), 6.77 (s, 1H), 5.31 (s, 2H), 3.75 (t, 1H, J 8.4), 2.90–2.86 (m, 2H), 2.36–1.88 (m, 6H), 1.73–1.71 (m, 1H), 1.55–1.21 (m, 7H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.0, 147.4, 138.5, 133.8, 126.7, 125.8, 120.8, 114.9, 112.3, 82.0, 61.9, 50.2, 44.1, 43.4, 38.9, 36.8, 30.8, 30.0, 27.3, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3586, 3083, 2919, 2872, 1595, 1572, 1519, 1501, 1339, 1254, 1238, 1029, 980, 852, 797, 749, 685, 508; EIMS m/z (%) 474 (M⁺, 12), 429 (8), 293 (4), 272 (100), 253 (4), 213 (19), 186 (8), 175 (38), 145 (20), 129 (39), 107 (8), 76 (11); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₀O₄N₄ 474.2262, found 474.2261.

1-(4-(4-(((17-Hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)­phenyl)­ethan-1-one (33)

Yellow solid, mp 193–194 °C, yield (80%); ¹H NMR (400 MHz, CDCl₃) δ = 8.26 (s, 1H), 8.16 (d, 2H, J 8.4), 7.92 (d, 2H, J 8.4), 7.26 (d, 1H, J 8.8), 6.86 (d, 1H, J 8.0), 6.78 (s, 1H), 5.30 (s, 2H), 3.75 (t, 1H, J 8.0), 2.86–2.85 (m, 2H), 2.68 (s, 3H), 2.35–1.89 (m, 5H), 1.73–1.18 (m, 9H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 196.7, 156.1, 138.4, 137.0, 133.7, 130.3, 126.6, 120.3, 114.9, 112.3, 82.0, 62.0, 50.2, 44.1, 43.4, 38.9, 36.8, 30.7, 29.9, 27.3, 26.8, 26.4, 23.3, 11.2; FT-IR (cm^–1) 3375, 2922, 2867, 1684, 1604, 1498, 1443, 1232, 1049, 1023, 830, 591; EIMS m/z (%) 471 (M⁺, 6), 381 (6), 270 (78), 252 (33), 213 (26), 185 (38), 172 (52), 132 (83), 95 (36), 81 (49), 69 (100); HRMS (ESI) [m]⁺ calcd for C₂₉H₃₃O₃N₃ 471.2516, found 471.2518.

4-(4-(((17-Hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)­benzenesulfonamide (34)

White solid, mp 218–221 °C, yield (79%); ¹H NMR (600 MHz, DMSO) δ = 9.03 (s, 1H), 8.15–8.13 (dt, 2H, J 6.0, 0.6), 8.04–8.01 (dt, 2H, J 6.0, 0.6), 7.53 (s, 2H), 7.19 (d, 1H, J 6.0), 6.82–6.80 (dd, 1H, J 6.0, 0.6), 6.75 (d, 1H, J 0.6), 5.19 (s, 2H), 4.49 (d, 1H, J 0.6), 3.52–3.33 (m, 1H), 2.79–2.49 (m, 2H), 2.28–1.10 (m, 13H), 0.66 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.7, 144.5, 143.8, 138.5, 137.5, 132.7, 127.4, 126.2, 122.8, 120.3, 114.4, 112.1, 80.0, 60.8, 49.5, 43.5, 42.7, 38.5, 36.5, 29.8, 29.2, 26.8, 25.9, 22.7, 11.2; FT-IR (cm^–1) 3511, 3278, 3099, 2921, 2867, 1595, 1497, 1346, 1234, 1166, 1054, 1022, 836, 615, 548; EIMS m/z (%) 508 (M⁺, 5), 474 (10), 398 (6), 309 (12), 272 (100), 254 (10), 209 (46), 172 (42), 160 (28), 130 (62), 92 (13), 77 (18); HRMS (ESI) [m]⁺ calcd for C₂₇H₃₂O₄N₄S 508.2139, found 508.2138.

13-Methyl-3-((1-(4-(trifluoromethyl)­phenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (35)

White solid, mp 186–188 °C, yield (63%); ¹H NMR (400 MHz, DMSO) δ = 9.07 (s, 1H), 8.19 (d, 2H, J 8.8), 8.01 (d, 2H, J 8.8), 7.20 (d, 1H, J 8.8), 6.83–6.80 (dd, 1H, J 8.8, 2.0), 6.76 (s, 1H), 5.19 (s, 2H), 3.55 (t, 1H, J 8.4), 2.78–2.75 (m, 2H), 2.28–1.79 (m, 5H), 1.62–1.15 (m, 9H), 0.66 (s, 3H); ¹³C­{¹H} NMR (100 MHz, DMSO) δ = 155.7, 144.6, 137.6, 132.8, 129.7, 127.28, 127.24, 126.2, 122.9, 120.6, 120.2, 119.7, 114.4, 112.2, 80.0, 60.8, 49.5, 43.5, 42.8, 36.5, 31.3, 29.8, 29.2, 26.8, 26.0, 22.7, 11.2; FT-IR (cm^–1)­3376, 2932, 2870, 1611, 1523, 1497, 1322, 1246, 1164, 1125, 1067, 1021, 845, 595; EIMS m/z (%) 497 (M⁺, 14), 477 (16), 441 (10), 316 (8), 303 (12), 272 (100), 198 (73), 172 (14), 145 (31), 91 (4); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₀O₂N₃F₃ 497.2285, found 497.2284.

2-(4-(((17-Hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)­benzoic Acid (36)

Pale yellow solid, mp 233–235 °C, yield (90%); ¹H NMR (600 MHz, DMSO) δ = 8.58 (s, 1H), 7.92 (d, 1H, J 6.6), 7.74 (t, 1H, J 7.8), 7.68 (t, 1H, J 7.2), 7.61 (d, 1H, J 7.2), 7.19 (d, 1H, J 9.0), 6.81–6.80 (dd, 1H, J 8.4, 2.4), 6.75 (d, 1H, J 1.8), 5.13 (s, 2H), 4.51 (s, 1H), 2.78–2.77 (m, 2H), 2.27–1.57 (m, 8H), 1.38–1.09 (m, 7H), 0.65 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 155.9, 143.1, 137.6, 132.8, 132.5, 130.6, 130.1, 126.6, 126.3, 126.0, 114.5, 112.3, 80.1, 67.1, 60.9, 49.6, 43.6, 42.9, 38.6, 36.6, 29.9, 29.3, 26.9, 26.1, 25.2, 22.8, 11.3; FT-IR (cm^–1) 3400, 3085, 2924, 2870, 2616, 1686, 1607, 1496, 1458, 1281, 1249, 1134, 1054, 1018, 756, 645, 570; EIMS m/z (%) 473 (M⁺, 19), 426 (4), 398 (10), 309 (3), 272 (96), 254 (10), 213 (41), 172 (32), 146 (42), 130 (100), 77 (34); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₁O₄N₃ 473.2309, found 473.2309.

13-Methyl-3-((1-(naphthalen-1-yl)-1H-1,2,3-triazol-4-yl)­methoxy)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (37)

Yellow oil, yield (84%); ¹H NMR (400 MHz, CDCl₃) δ = 8.06–8.03 (m, 1H), 8.02 (s, 1H), 7.99 (d, 1H, J 8.4), 7.66–7.54 (m, 5H), 7.27 (d, 1H, J 8.8), 6.89–6.86 (dd, 1H, J 8.8, 2.8), 6.81 (d, 1H, J 2.8), 5.38 (s, 2H), 3.75 (t, 1H, J 8.4), 2.90–2.87 (m, 2H), 2.35–2.11 (m, 3H), 1.99–1.71 (m, 3H), 1.53–1.20 (m, 8H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.3, 144.7, 138.4, 134.3, 133.8, 133.6, 130.6, 128.6, 128.4, 128.1, 127.3, 126.6, 125.5, 125.1, 123.7, 122.4, 115.0, 112.5, 82.0, 62.3, 50.2, 44.1, 43.4, 39.0, 36.9, 30.7, 30.0, 27.4, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3394, 2924, 2867, 1728, 1602, 1496, 1379, 1233, 1020, 800, 771, 570; EIMS m/z (%) 479 (M⁺, 4), 450 (6), 423 (5), 280 (4), 272 (6), 213 (2), 180 (100), 152 (9), 127 (16), 77 (2); HRMS (ESI) [m]⁺ calcd for C₃₁H₃₃O₂N₃ 479.2567, found 479.2567.

3-((1-Benzyl-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (38)

White solid, mp 182–184 °C, yield (71%); ¹H NMR (400 MHz, CDCl₃) δ = 7.56 (s, 1H), 7.42–7.38 (m, 3H), 7.31–7.29 (m, 2H), 7.22 (d, 1H, J 8.8), 6.79–6.76 (dd, 1H, J 8.8, 2.8), 6.71 (d, 1H, J 2.8), 5.56 (s, 2H), 5.20 (s, 2H), 3.75 (t, 1H, J 8.4), 2.87–2.83 (m, 2H), 2.41–2.11 (m, 5H), 1.99–1.87 (m, 2H), 1.71–1.70 (m, 1H), 1.52–1.20 (m, 6H), 0.78 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.2, 144.9, 138.3, 134.3, 133.6, 129.4, 129.1, 128.4, 126.6, 122.9, 114.9, 112.4, 82.1, 62.1, 54.7, 50.2, 44.1, 43.4, 39.0, 36.9, 30.8, 29.9, 27.4, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3553, 3131, 2943, 2907, 2870, 1603, 1498, 1461, 1382, 1311, 1229, 1125, 1068, 1050, 1013, 731, 708, 570; EIMS m/z (%) 443 (M⁺, 28), 414 (4), 386 (3), 324 (6), 272 (28), 213 (8), 172 (6), 144 (100), 117 (7), 91 (90), 65 (4); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₃O₂N₃ 443.2567, found 443.2567.

3-((1-(3,4-Dimethylphenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (39)

Yellow solid, mp 189–191 °C, yield (78%); ¹H NMR (400 MHz, CDCl₃) δ = 8.19 (s, 1H), 7.55 (s, 1H), 7.46 (d, 1H, J 8.0), 7.28 (d, 1H, J 8.0), 7.24 (d, 1H, J 7.2), 6.86 (d, 1H, J 8.0), 6.78 (s, 1H), 5.27 (s, 2H), 3.75 (t, 1H, J 8.4), 2.87–2.85 (m, 2H), 2.36 (s, 3H), 2.33 (s, 3H), 2.30–1.70 (m, 6H), 1.53–1.18 (m, 7H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.1, 138.5, 138.2, 137.7, 130.7, 126.5, 121.8, 118.0, 114.8, 112.3, 81.9, 62.1, 50.1, 44.0, 43.3, 38.9, 38.8, 36.8, 30.6, 29.9, 27.3, 26.4, 23.2, 20.0, 19.6, 11.2; FT-IR (cm^–1) 3274, 2926, 2876, 1692, 1604, 1495, 1450, 1282, 1229, 1045, 1020, 815, 729, 574; EIMS m/z (%) 457 (M⁺, 8), 401 (10), 310 (6), 272 (8), 263 (3), 213 (2), 158 (100), 105 (6), 78 (9); HRMS (ESI) [m]⁺ calcd for C₂₉H₃₅O₂N₃ 457.2724, found 457.2724.

3-((1-Mesityl-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (40)

White solid, mp 199–200 °C, yield (77%); ¹H NMR (400 MHz, CDCl₃) δ = 7.64 (s, 1H), 7.24 (d, 1H, J 8.4), 7.00 (s, 2H), 6.84–6.81 (dd, 1H, J 8.8, 2.8), 6.75 (d, 1H, J 2.8), 5.31 (s, 2H), 3.75 (t, 1H, J 8.4), 2.87–2.84 (m, 2H), 2.37 (s, 3H), 2.35–1.99 (m, 5H), 1.97 (s, 6H), 1.91–1.20 (m, 9H), 0.80 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.3, 144.6, 140.2, 138.3, 135.2, 133.5, 129.2, 126.5, 124.6, 115.2, 112.6, 82.0, 62.6, 50.2, 44.1, 43.4, 39.0, 36.9, 30.7, 29.9, 27.4, 26.4, 23.3, 21.3, 17.4, 11.2; FT-IR (cm^–1) 3493, 3141, 2919, 2864, 1709, 1605, 1495, 1380, 1248, 1225, 1049, 1018, 854, 802, 579; EIMS m/z (%) 471 (M⁺, 8), 400 (4), 277 (5), 172 (100), 157 (13), 119 (5), 91 (6); HRMS (ESI) [m]⁺ calcd for C₃₀H₃₇O₂N₃ 471.2880, found 471.2880.

3-((1-(4-Iodo-2-methylphenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (41)

Yellow oil, yield (76%); ¹H NMR (400 MHz, CDCl₃) δ = 7.83 (s, 1H), 7.75 (d, 1H, J 2.4), 7.67–7.65 (dd, 1H, J 8.4, 2.0), 7.23 (d, 1H, J 8.8), 7.08 (d, 1H, J 8.4), 6.83–6.81 (dd, 1H, J 8.4, 2.0), 6.75 (s, 1H), 5.27 (s, 2H), 3.74 (t, 1H, J 8.8), 2.86–2.85 (m, 2H), 2.2.35–2.08 (m, 6H), 1.97–1.87 (m, 2H), 1.74–1.27 (m, 9H), 0.79 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 156.0, 140.4, 138.1, 136.1, 136.0, 135.7, 133.5, 127.4, 126.4, 114.9, 112.2, 95.6, 81.7, 62.0, 50.0, 43.9, 43.2, 38.8, 36.7, 30.5, 29.8, 27.2, 26.3, 23.1, 17.7, 11.2; FT-IR (cm^–1) 3377, 2922, 2868, 1702, 1605, 1494, 1445, 1280, 1232, 1131, 1042, 1021, 815, 731, 561; EIMS m/z (%) 569 (M⁺, 8), 540 (13), 513 (8), 448 (4), 388 (8), 375 (12), 272 (84), 244 (6), 213 (16), 159 (20), 143 (100), 115 (21); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₂O₂N₃I 569.1534, found 569.1529.

3-((1-(3,5-Bis­(trifluoromethyl)­phenyl)-1H-1,2,3-triazol-4-yl)­methoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-17-ol (42)

White solid, mp 193–195 °C, yield (88%); ¹H NMR (600 MHz, CDCl₃) δ = 8.25 (s, 2H), 7.94 (s, 1H), 7.22 (d, 1H, J 7.8), 6.81 (d, 1H, J 7.8), 6.73 (s, 1H), 5.27 (s, 2H), 3.71 (t, 1H, J 8.4), 2.86–2.83 (m, 2H), 2.31–1.85 (m, 5H), 1.69–1.16 (m, 10H), 0.76 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 156.0, 138.5, 134.3, 134.0, 133.9, 133.8, 126.7, 123.6, 122.4, 121.8, 120.7, 114.9, 112.3, 82.1, 50.2, 44.1, 43.4, 39.0, 36.9, 30.8, 30.0, 27.4, 26.5, 23.3, 11.2; FT-IR (cm^–1) 3360, 2915, 2870, 1608, 1576, 1495, 1355, 1281, 1175, 1136, 1039, 890, 824, 682; EIMS m/z (%) 565 (M⁺, 9), 524 (6), 369 (8), 341 (6), 286 (10), 266 (100), 246 (12), 213 (34), 160 (12), 133 (12), 91 (8); HRMS (ESI) [m]⁺ calcd for C₂₉H₂₉O₂N₃F₆ 565.2158, founf 565.2159.

2-(4-(((17-Hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)-N-(3-nitrophenyl)­benzamide (43)

Pale yellow solid, mp 268–270 °C, yield (89%); ¹H NMR (600 MHz, DMSO) δ = 10.99 (s, 1H), 8.76 (s, 1H), 8.64 (t, 1H, J 1.8), 7.99–7.93 (dq, 1H, J 8.4, 1.8), 7.88–7.87 (dd, 1H, J 7.8, 1.2), 7.84–7.82 (dd, 1H, J 7.2, 1.2), 7.79–7.66 (m, 3H), 7.61 (t, 1H, J 8.4), 7.15 (d, 1H, J 9.0), 6.77–6.75 (dd, 1H, J 8.4, 2.4), 6.72 (d, 1H, J 1.8), 5.12 (s, 2H), 4.56 (d, 1H, J 4.8), 3.60 (t, 2H, J 4.2), 2.75–2.74 (m, 2H), 1.87–1.09 (m, 12H), 0.66 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 165.4, 155.9, 147.9, 143.5, 140.0, 137.6, 134.0, 132.7, 132.1, 131.6, 130.2, 129.7, 129.2, 126.2, 125.7, 125.4, 125.2, 118.4, 114.5, 113.8, 112.2, 80.1, 67.1, 60.9, 49.6, 43.6, 42.8, 38.6, 36.6, 29.9, 29.3, 26.9, 26.0, 25.2, 22.8, 11.3; FT-IR (cm^–1) 3274, 2921, 2867, 1670, 1604, 1528, 1497, 1429, 1350, 1249, 1050, 1022, 760, 737, 671, 529; EIMS m/z (%) 593 (M⁺, 8), 498 (2), 425 (5), 389 (6), 293 (10), 272 (100), 213 (54), 172 (40), 160 (44), 120 (70), 92 (36), 76 (18); HRMS (ESI) [m]⁺ calcd for C₃₄H₃₅O₅N₅ 593.2456, found 593.2455.

2-(4-(((17-Hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)-N-(4-(trifluoromethyl)­phenyl)­benzamide (44)

White solid, mp 251–254 °C, yield (92%); ¹H NMR (600 MHz, DMSO) δ = 10.83 (s, 1H), 8.61 (s, 1H), 7.80 (d, 1H, J 7.2), 7.75–7.65 (m, 5H), 7.65 (d, 2H, J 8.4), 7.15 (d, 1H, J 8.4), 6.76 (d, 1H, J 8.4), 6.70 (s, 1H), 5.11 (s, 2H), 4.55 (d, 1H, J 4.8), 3.59 (s, 2H), 2.75–2.74 (m, 2H), 2.25–1.07 (m, 12H), 0.65 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 165.7, 156.2, 143.8, 142.8, 138.0, 134.4, 133.1, 132.7, 131.8, 130.0, 129.5, 126.6, 126.45, 126.42, 125.8, 125.6, 120.1, 114.9, 112.6, 80.5, 67.4, 61.2, 49.9, 43.9, 43.2, 39.0, 37.0, 30.3, 29.6, 27.2, 26.4, 25.5, 23.2, 11.6; FT-IR (cm^–1) 3273, 2925, 2868, 1657, 1604, 1527, 1497, 1407, 1320, 1251, 1115, 1065, 1018, 831, 760, 508; EIMS m/z (%) 616 (M⁺, 12), 575 (10), 472 (14), 425 (26), 356 (48), 317 (93), 289 (82), 272 (78), 247 (19), 172 (46), 145 (100), 77 (19); HRMS (ESI) [m]⁺ calcd for C₃₅H₃₅O₃N₄F₃ 616.2656, found 616.2654.

2-(4-(((17-Hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1H-1,2,3-triazol-1-yl)-N-(naphthalen-1-yl)­benzamide (45)

White solid, mp 217–219 °C, yield (86%); ¹H NMR (600 MHz, DMSO) δ = 10.51 (s, 1H), 8.62 (s, 1H), 7.95–7.91 (m, 3H), 7.81 (d, 1H, J 8.4), 7.75–7.71 (m, 3H), 7.57 (d, 1H, J 7.8), 7.53–7.48 (m, 3H), 7.14 (d, 1H, J 8.4), 6.79–6.77 (dd, 1H, J 8.4, 2.4), 6.72 (d, 1H, J 2.4), 5.14 (s, 2H), 4.57 (d, 1H, J 4.8), 3.60–3.58 (m, 4H), 2.74–2.72 (m, 2H), 2.24–1.08 (m, 10H), 0.66 (s, 3H); ¹³C­{¹H} NMR (150 MHz, DMSO) δ = 165.9, 155.9, 143.5, 137.6, 134.1, 133.7, 133.1, 132.8, 131.0, 129.7, 129.2, 128.6, 128.0, 126.3, 126.2, 126.1, 126.0, 125.57, 125.53, 125.2, 123.2, 122.9, 114.5, 112.2, 80.1, 67.1, 61.0, 49.6, 43.6, 42.9, 38.6, 36.6, 29.9, 29.3, 26.9, 26.1, 25.2, 22.8, 11.3; FT-IR (cm^–1) 3410, 2936, 2868, 1678, 1607, 1538, 1497, 1344, 1254, 1131, 794, 772, 600, 593; EIMS m/z (%) 598 (M⁺, 12), 558 (4), 449 (3), 425 (8), 323 (14), 298 (20), 286 (24), 272 (100), 255 (13), 213 (28), 159 (30), 146 (35), 120 (54), 77 (12); HRMS (ESI) [m]⁺ calcd for C₃₈H₃₈O₃N₄ 598.2938, found 598.2936.

4-(((13-Methyl-17-octyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-3-yl)­oxy)­methyl)-1-(4-(trifluoromethyl)­phenyl)-1H-1,2,3-triazole (46)

White solid, mp 174–176 °C, yield (69%); ¹H NMR (600 MHz, CDCl₃) δ = 8.06 (s, 1H), 7.80 (d, 2H, J 8.4), 7.66 (d, 2H, J 8.4), 3.43–3.37 (m, 1H), 2.41 (t, 2H, J 2.4), 2.02–0.87 (m, 41H), 0.69 (s, 3 H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 144.4, 129.1, 126.64, 126.61, 126.56, 126.50, 119.9, 119.5, 79.5, 72.0, 61.7, 56.9, 56.3, 50.3, 42.5, 40.0, 39.7, 39.2, 37.4, 37.0, 36.4, 36.0, 32.1, 31.8, 28.4, 28.2, 24.5, 24.0, 23.0, 22.7, 21.3, 19.6, 18.9, 12.0; FT-IR (cm^–1) 3351, 2932, 2867, 2410, 2129, 1657, 1603, 1530, 1501, 1408, 1322, 1258, 1066, 1018, 837, 759, 634, 594; EIMS m/z (%) 611 (M⁺, 5), 384 (64), 367 (100), 352 (78), 275 (74), 255 (54), 213 (74), 145 (88), 107 (72), 81 (89); HRMS (ESI) [m]⁺ calcd for C₃₇H₅₂ON₃F₃ 611.4057, found 611.4058.

13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl 1-Phenyl-1H-1,2,3-triazole-4-carboxylate (47)

White solid, mp 264–265 °C, yield (91%); ¹H NMR (600 MHz, CDCl₃) δ = 8.68 (s, 1H), 7.83 (d, 2H, J 7.8), 7.62 (t, 2H, J 7.4), 7.55 (t, 1H, J 7.4), 7.38 (d, 1H, J 8.4), 7.07–7.06 (dd, 1H, J 8.4, 2.4), 7.03 (s, 1H), 2.98–2.97 (m, 2H), 2.57–2.00 (m, 6H), 1.69–1.49 (m, 7H), 0.82 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 159.4, 148.3, 138.4, 138.1, 136.5, 130.2, 129.9, 126.7, 126.5, 121.8, 121.1, 118.9, 50.6, 48.17, 44.4, 44.3, 38.1, 36.0, 31.7, 29.9, 29.6, 26.5, 25.9, 21.8, 14.0; FT-IR (cm^–1) 3148, 2930, 2865, 2114, 1728, 1597, 1544, 1491, 1244, 1203, 1152, 1027, 907, 758, 683; EIMS m/z (%) 441 (M⁺, 40),397 (55), 369 (39), 270 (62), 213 (15), 185 (22), 172 (59), 144 (100), 116 (44), 77 (34); HRMS (ESI) [m]⁺ calcd for C₂₇H₂₇O₃N₃ 441.2047, found 441.2047.

13-Methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta­[a]­phenanthren-3-yl 1-(4-(Trifluoromethyl)­phenyl)-1H-1,2,3-triazole-4-carboxylate (48)

White solid, mp 195–197 °C, yield (65%); ¹H NMR (400 MHz, CDCl₃) δ = 8.79 (s, 1H), 8.01 (d, 2H, J 8.4), 7.88 (d, 2H, J 8.8), 7.36 (d, 1H, J 8.4), 7.05–7.03 (dd, 1H, J 8.4, 2.8), 7.00 (d, 1H, J 2.4), 2.96–2.94 (m, 2H), 2.57–1.97 (m, 6H), 1.69–1.27 (m, 7H), 0.94 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 159.1, 148.1, 140.7, 138.8, 138.4, 138.1, 127.55, 127.51, 126.7, 126.4, 121.6, 121.0, 118.8, 50.6, 48.1, 44.3, 38.1, 36.0, 31.7, 29.5, 26.4, 25.9, 21.7, 14.0; FT-IR (cm^–1) 3143, 2929, 1764, 1737, 1616, 1545, 1495, 1329, 1243, 1220, 1160, 1112, 1070, 1024, 843, 761, 592; EIMS m/z (%) 509 (M⁺, 82), 465 (86), 437 (19), 270 (24), 212 (100), 184 (45), 145 (42), 133 (14), 97 (9); HRMS (ESI) [m]⁺ calcd for C₂₈H₂₆O₃N₃F₃ 509.1921, found 509.1923.

2,3,4,7,8,9,10,11,12,13,14,15,16,17-Tetradecahydro-1H-cyclopenta­[a]­phenanthren-3-yl 1-(4-Chlorophenyl)-1H-1,2,3-triazole-4-carboxylate (49)

White solid, mp 218–220 °C, yield (85%); ¹H NMR (400 MHz, CDCl₃) δ = 8.55 (s, 1H), 7.76 (d, 2H, J 8.8), 7.57 (d, 2H, J 8.8), 5.49 (d, 1H, J 4.8), 5.03–4.95 (m, 1H), 2.60–2.46 (m, 3H), 2.18–1.87 (m, 7H), 1.73–1.22 (m, 9H), 1.11 (s, 3H), 0.92 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 162.5, 139.8, 135.6, 130.3, 122.5, 122.2, 75.3, 51.9, 50.3, 47.7, 38.2, 37.1, 36.9, 36.0, 31.67, 31.60, 31.0, 27.8, 22.0, 20.5, 19.5, 13.7; FT-IR (cm^–1) 3143, 3075, 2946, 2851, 1743, 1720, 1546, 1502, 1261, 1248, 1154, 1093, 1035, 1007, 827, 768, 532; EIMS m/z (%) 493 (M⁺, 26), 270 (100), 255 (18), 224 (24), 199 (7), 178 (9), 121 (28), 105 (10), 79 (6); HRMS (ESI) [m]⁺ calcd for C₂₈H₂₃O₃N₃Cl 493.2127, found 493.2125.

10,13-Dimethyl-17-oxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-3-yl 1-(4-Acetylphenyl)-1H-1,2,3-triazole-4-carboxylate (50)

White solid, mp 194–196 °C, yield (87%); ¹H NMR (400 MHz, CDCl₃) δ = 8.61 (s, 1H), 8.19 (d, 2H, J 8.8), 7.94–7.92 (dd, 2H, J 8.8, 2.0), 5.50 (d, 1H, J 4.4), 5.03–4.95 (m, 1H), 2.69 (s, 3H), 2.60–2.46 (m, 3H), 2.17–1.85 (m, 6H), 1.73–1.23 (m, 7H), 1.12 (s, 3H), 0.92 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 196.6, 160.5 139.8, 137.7, 130.4, 122.5, 120.7, 75.4, 51.9, 50.3, 47.7, 38.2, 37.1, 37.0, 36.0, 31.69, 31.62, 31.0, 27.9, 26.9, 22.1, 20.5, 19.5, 13.7; FT-IR (cm^–1) 3125, 2942, 2860, 1715, 1683, 1604, 1541, 1359, 1263, 1161, 1035, 837, 774, 590; EIMS m/z (%) 501 (M⁺, 10), 270 (100), 255 (26), 232 (21), 159 (10), 145 (14), 121 (40), 91 (24), 79 (14); HRMS (ESI) [m]⁺ calcd for C₃₀H₃₅O₄N₃ 501.2673, found 501.2671.

17-Acetyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-3-yl 1-Phenyl-1H-1,2,3-triazole-4-carboxylate (51)

White solid, mp 255–257 °C, yield (90%); ¹H NMR (600 MHz, CDCl₃) δ = 8.54 (s, 1H), 7.79 (d, 2H, J 7.8), 7.59 (t, 2H, J 8.4), 7.52 (t, 1H, J 7.2), 5.47 (d, 1H, J 3.6), 5.00 (s, 1H), 2.59–2.52 (m, 3H), 2.44–2.03 (m, 7H), 1.98–1.86 (m, 2H), 1.76–1.46 (m, 7H), 1.30–1.06 (m, 7H), 0.67 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 209.7, 160.5, 139.6, 130.1, 129.7, 122.9, 121.1, 75.3, 63.9, 57.0, 50.1, 44.2, 39.0, 38.2, 37.2, 36.8, 32.05, 32.01, 31.7, 31.4, 27.9, 24.7, 23.0, 21.2, 19.5, 13.4; FT-IR (cm^–1) 3129, 2922, 2892, 2853, 1717, 1687, 1539, 1355, 1261, 1247, 1153, 775, 759, 686, 595; EIMS m/z (%) 487 (M⁺, 8), 368 (11), 298 (100), 283 (22), 236 (16), 190 (54), 121 (22), 97 (38), 83 (45), 69 (58); HRMS (ESI) [m]⁺ calcd for C₃₀H₃₇O₃N₃ 487.2829, found 487.2828.

17-Acetyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-3-yl 1-(4-(Trifluoromethyl)­phenyl)-1H-1,2,3-triazole-4-carboxylate (52)

Pale yellow solid, mp 240–242 °C, yield (65%); ¹H NMR (600 MHz, CDCl₃) δ = 8.59 (s, 1H), 7.89 (d, 2H, J 7.8), 7.67 (d, 2H, J 8.4), 5.47 (s, 1H), 5.00–4.98 (m, 1H), 2.59–2.52 (m, 3H), 2.44–2.03 (m, 7H), 1.98–1.86 (m, 2H), 1.76–1.46 (m, 7H), 1.30–1.06 (m, 7H), 0.67 (s, 3H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 209.8, 164.5, 141.6, 129.4, 126.62, 126.60, 126.57, 125.55, 120.8, 120.2, 75.6, 63.8, 57.0, 50.0, 44.2, 38.9, 38.2, 37.1, 36.8, 32.0, 31.9, 31.7, 27.9, 24.6, 23.0, 21.2, 19.5, 13.4; FT-IR (cm^–1) 2941, 1704, 1682, 1605, 1532, 1321, 1258, 1159, 1112, 1066, 1033, 841, 593, 507; EIMS m/z (%) 554 (M⁺, 4), 451 (3), 331 (9), 298 (56), 269 (18), 213 (26), 159 (33), 144 (100), 91 (46); HRMS (ESI) [m]⁺ calcd for C₃₁H₃₆O₃N₃F₃ 555.2803, found 555.2806.

10,13-Dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-17-yl 1-Phenyl-1H-1,2,3-triazole-4-carboxylate (53)

White solid, mp 131–133 °C, yield (95%); ¹H NMR (600 MHz, CDCl₃) δ = 8.49 (s, 1H), 7.79–7.77 (dt, 2H, J 7.2, 1.2), 7.58 (t, 2H, J 7.8), 7.53–7.50 (td, 1H, J 7.2, 1.2), 5.76 (s, 1H), 4.97 (t, 1H, J 7.8), 2.45–2.32 (m, 5H), 2.06–2.04 (m, 3H), 1.94–1.89 (m, 2H), 1.78–1.61 (m, 5H), 1.48–1.17 (m, 10H), 1.02–1.00 (m, 4H); ¹³C­{¹H} NMR (150 MHz, CDCl₃) δ = 199.6, 171.0, 160.8, 141.1, 136.6, 130.1, 129.7, 125.5, 124.1, 121.0, 83.6, 53.8, 53.6, 50.5, 43.1, 38.8, 36.8, 35.9, 35.6, 34.1, 32.9, 31.6, 31.4, 27.7, 23.7, 20.7, 17.6, 12.4; FT-IR (cm^–1) 3135, 2942, 2853, 1735, 1657, 1503, 1232, 1163, 1041, 773, 738, 692; EIMS m/z (%) 459 (M⁺, 100), 415 (8), 308 (4), 288 (29), 271 (62), 253 (48), 188 (19), 172 (52), 116 (5); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₃O₃N₃ 459.2516, found 459.2514.

10,13-Dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-17-yl 1-(4-Chlorophenyl)-1H-1,2,3-triazole-4-carboxylate (54)

White solid, mp 197–198 °C, yield (78%); ¹H NMR (400 MHz, CDCl₃) δ = 8.59 (s, 1H), 7.76 (d, 2H, J 8.8), 7.57 (d, 2H, J 8.4), 5.77 (s, 1H), 5.00 (t, 1H, J 8.0), 2.43–2.33 (m, 5H), 2.10–2.07 (m, 1H), 1.95–1.29 (m, 13H), 1.23 (s, 3H), 1.03 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 171.0, 160.9, 135.6, 135.1, 130.4, 125.7, 124.2, 122.2, 83.7, 53.9, 50.5, 43.1, 38.8, 36.8, 35.9, 35.6, 32.9, 31.6, 27.7, 23.7, 20.7, 17.6, 12.4; FT-IR (cm^–1) 3134, 2939, 2853, 1709, 1674, 1542, 1502, 1344, 1269, 1169, 1035, 842, 775, 516; EIMS m/z (%) 493 (M⁺, 14), 288 (9), 253 (10), 229 (4), 195 (6), 178 (100), 147 (17), 133 (8), 91 (11); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₂O₃N₃Cl 493.2127, found 493.2127.

10,13-Dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1cyclopenta­[a]­phenanthren-17-yl 1-(4-Bromophenyl)-1H-1,2,3-triazole-4-carboxylate (55)

White solid, mp 185–187 °C, yield (84%); ¹H NMR (400 MHz, CDCl₃) δ = 8.58 (s, 1H), 7.72 (d, 2H, J 8.8), 7.70 (d, 2H, J 9.2), 5.77 (s, 1H), 4.99 (t, 1H, J 8.0), 2.43–2.30 (m, 5H), 2.09–2.06 (m, 1H), 1.94–1.29 (m, 13H), 1.22 (s, 3H), 1.02 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 171.0, 160.7, 135.9,133.3, 133.0, 123.5, 122.4, 83.7, 53.8, 50.5, 43.1, 38.8, 36.8, 35.9, 35.6, 32.9, 31.6, 31.4, 27.7, 23.7, 20.7, 17.6, 12.4; FT-IR (cm^–1) 3133, 2937, 2857, 1708, 1672, 1499, 1268, 1168, 1034, 839, 774, 513; EIMS m/z (%) 537 (M⁺, 22), 288 (12), 253 (13), 222 (100), 197 (8), 147 (21), 133 (10), 91 (14), 79 (10); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₂O₃N₃Br 537.1622, found 537.1620.

10,13-Dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta­[a]­phenanthren-17-yl 1-(Naphthalen-1-yl)-1H-1,2,3-triazole-4-carboxylate (56)

White solid, mp 175–177 °C, yield (67%); ¹H NMR (400 MHz, CDCl₃) δ = 8.44 (s, 1H), 8.08–8.06 (m, 1H), 7.98 (d, 1H, J 7.6), 7.64–7.57 (m, 5H), 5.74 (s, 1H), 5.01–4.97 (td, 1H, J 8.0, 1.2), 2.43–2.32 (m, 5H), 2.06–2.02 (m, 1H), 1.97–1.26 (m, 13H), 1.21 (s, 3H), 1.02 (s, 3H); ¹³C­{¹H} NMR (100 MHz, CDCl₃) δ = 199.6, 171.0, 160.8, 140.5, 134.2, 133.0, 131.1, 130.0, 128.5, 128.4, 128.3, 127.4, 125.1, 124.1, 123.9, 122.0, 83.6, 53.8, 50.4, 43.1, 38.7, 36.8, 35.8, 35.5, 34.0, 32.8, 31.6, 27.7, 23.7, 20.7, 17.5, 12.4; FT-IR (cm^–1) 3137, 2936, 2847, 1714, 1668, 1538, 1296, 1245, 1153, 1030, 800, 771, 569; EIMS m/z (%) 509 (M⁺, 5), 481 (10), 238 (6), 211 (10), 194 (100), 167 (38), 127 (8), 91 (6); HRMS (ESI) [m]⁺ calcd for C₂₈H₃₅O₃N₃ 509.2673, found 509.2671.

Antibacterial Activity

The Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli) were inoculated overnight in LB broth medium at 37 °C in the shaker. The cultures were then diluted 10⁵-fold using LB broth and seeded in 96-well plate (200 μL/well) and treated with 2 μL of 10, 1, and 0.1 mM stock solutions of each compound prepared in DMSO, negative control (DMSO), and 2 μL of 10, 1, and 0.1 mg/mL of the positive control (kanamycin). The plate was then incubated at 37 °C overnight (16–20h/static). Following the incubation period, OD600 was measured using SpectraMax plate reader. The hits identified from the initial screen were validated by retesting the active compounds in a dose–response format (2-fold serial dilution starting at 100 μM final concentration of test compounds and 100 μg/mL (final concentration) of the positive control kanamycin). Each treatment was done in triplicate. The minimum inhibitory concentration (MIC) was reported as the lowest concentration of compound at which no observed bacterial growth.

To assess the inhibition of biofilm, the Gram-positive bacteria (S. aureus) was inoculated overnight in LB broth medium at 37 °C in the shaker. The culture was then diluted 10⁵-fold using LB broth and seeded in 96-well plates (200 μL/well) and treated in dose response format (2-fold serial dilution) with the 0.1–100 μM (final concentration) for compounds 46, 47, and 52; and 0.1–100 μg/mL (final concentration) for kanamycin. The plate was then incubated at 37 °C overnight (16–20h/static). Following the incubation period, the medium was discarded and the plates were washed with distilled water. The wells were then stained with 0.1% aqueous solution of crystal violet (100 μL per well) and allowed to stand at room temperature for 30 min. After that plates were washed again with distilled water and remaining stains were solubilized with 95% EtOH (200 μL per well). 125 μL from each well was transferred to a new plate and OD540 was measured using SpectraMax plate reader.

Anticancer Activity

The cells (MDA-MB-231) were propagated and maintained in advanced Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) supplemented with 5% fetal bovine serum (FBS; HyClone, Logan, UT, USA), 1% antibiotic–antimycotic (Invitrogen), 1% essential amino acids and 1% l-glutamine, at 37 °C humidified air and 5% CO₂.

The cells were seeded in 96-well plates (10,000 cells/well). After 24-h of incubation, the cells were treated with different concentrations of each test compound prepared in DMSO or solvent control (DMSO). Each treatment was done in triplicate. Following 48-h of incubation, cell viability was measured using 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide according to the manufacturer’s instructions (Promega).

The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsomega.5c11199.

• X-ray data of three samples; NMR and HRMS spectral data of steroid ethers and esters (1–7) and steroid-triazole (8–56); and single-crystal X-ray diffraction studies of 19, 38, and 47 (PDF)

The authors declare no competing financial interest.