The present invention provides novel dimeric and sulfonyl dimeric indolosesquiterpenes having antibacterial activities. These compounds can e.g. be isolated from a mangrove endophyte.

Indole terpenoids encompass a highly diverse group of natural products, including the anticancer agents vincristine and vinblastine, psychotropic agents such as lysergic acid derivatives, the aphrodisiac yohimbine, and the potassium channel blockers paxilline and lolitrem.

In light of the impressive number of known eukaryotic indole terpene metabolites, it is remarkable that only recently the first representatives of this group derived from bacteria were discovered, including oridamycin, xiamycin, indosespene, and sespenine (Takada 2010, Ding 2010, Ding 201 1). Xiamycin (1) was produced by two Streptomyces endophytes (HKI0576 and HKI0595) of widespread mangrove trees, Bruguiera gymnorrhiza and Kandelia candel, respectively (Ding 2010, Ding 2011).

From the two strains, the dimers of xiamycin have now been detected. Heterologous expression of the xiamycin biosynthesis gene cluster (Li 2012) in S. albus, follow-up fermentation and isolation yielded various N-N, and C-N dimers as well as sulfonyl bridged dimers of xiamycins (2a, 2b, 3a, 3b, 4, 5, 6, and 7). They all exhibit antibacterial activities and no cytotoxic effects indicating their use as antibacterial agents. Figure 1 shows the structures of xiamycin, bixiamycins and sulfonylbixiamycins.

The present invention provides a compound of formula (II)

X-A-Y (II) wherein A is a bond, a sulphur atom or a group of formula -SO- or -S0 ₂ -, wherein X has the following structure of formula (I) wherein one hydrogen atom is replaced by the bond to A or Y

(I) and wherein Y has the following structure of formula (I) wherein one hydrogen atom is replaced by the bond to A or X

(I) or a pharmaceutically acceptable salt, solvate or hydrate or a pharmaceutically acceptable formulation thereof.

Preferably, X is selected from the following structures:

Further preferably, Y is selected from the following structures:

(I) or a pharmaceutically acceptable salt, solvate or hydrate or a pharmaceutically acceptable formulation thereof. The present invention further provides pharmaceutical compositions comprising one or more compounds described herein or a pharmaceutically acceptable salt, solvate or hydrate thereof, optionally in combination with a pharmaceutically acceptable carrier.

It is a further object of the present invention to provide a compound as described herein or a pharmaceutical composition as defined herein for the preparation of a medicament for the treatment and/or prophylaxis of bacterial infections.

Examples of pharmacologically acceptable salts of sufficiently basic compounds are salts of physiologically acceptable mineral acids like hydrochloric, hydrobromic, sulfuric and phosphoric acid; or salts of organic acids like methanesulfonic, p-toluenesulfonic, lactic, acetic, trifluoroacetic, citric, succinic, fumaric, maleic and salicylic acid. Further, a sufficiently acidic compound may form alkali or earth alkali metal salts, for example sodium, potassium, lithium, calcium or magnesium salts; ammonium salts; or organic base salts, for example methylamine, dimethylamine, trimethylamine, triethylamine, ethylenediamine, ethanolamine, choline hydroxide, meglumin, piperidine, morpholine, tris-(2- hydroxyethyl)amine, lysine or arginine salts; all of which are also further examples of salts of the compounds described herein. The compounds described herein may be solvated, especially hydrated. The hydratization/hydration may occur during the process of production or as a consequence of the hygroscopic nature of the initially water free compounds. The solvates and/or hydrates may e.g. be present in solid or liquid form.

The therapeutic use of the compounds described herein, their pharmacologically acceptable salts, solvates and hydrates, respectively, as well as formulations and pharmaceutical compositions also lie within the scope of the present invention.

The pharmaceutical compositions according to the present invention comprise at least one compound described herein and, optionally, carrier substances and/or adjuvants.

The pharmaceutical composition may optionally comprise one or more further antibacterial compounds or may be administered in combination with one or more further antibacterial compounds. As mentioned above, therapeutically useful agents that contain compounds described herein, their solvates, salts or formulations are also comprised in the scope of the present invention. In general, the compounds described herein will be administered by using the known and acceptable modes known in the art, either alone or in combination with any other therapeutic agent.

For oral administration such therapeutically useful agents can be administered by one of the following routes: oral, e.g. as tablets, dragees, coated tablets, pills, semisolids, soft or hard capsules, for example soft and hard gelatine capsules, aqueous or oily solutions, emulsions, suspensions or syrups, parenteral including intravenous, intramuscular and subcutaneous injection, e.g. as an injectable solution or suspension, rectal as suppositories, by inhalation or insufflation, e.g. as a powder formulation, as microcrystals or as a spray (e.g. liquid aerosol), transdermal, for example via an transdermal delivery system (TDS) such as a plaster containing the active ingredient or intranasal. For the production of such tablets, pills, semisolids, coated tablets, dragees and hard, e.g. gelatine, capsules the therapeutically useful product may be mixed with pharmaceutically inert, inorganic or organic excipients as are e.g. lactose, sucrose, glucose, gelatine, malt, silica gel, starch or derivatives thereof, talc, stearinic acid or their salts, dried skim milk, and the like. For the production of soft capsules one may use excipients as are e.g. vegetable, petroleum, animal or synthetic oils, wax, fat, polyols. For the production of liquid solutions, emulsions or suspensions or syrups one may use as excipients e.g. water, alcohols, aqueous saline, aqueous dextrose, polyols, glycerin, lipids, phospholipids, cyclodextrins, vegetable, petroleum, animal or synthetic oils. Especially preferred are lipids and more preferred are phospholipids (preferred of natural origin; especially preferred with a particle size between 300 to 350 nm) preferred in phosphate buffered saline (pH = 7 to 8, preferred 7.4). For suppositories one may use excipients as are e.g. vegetable, petroleum, animal or synthetic oils, wax, fat and polyols. For aerosol formulations one may use compressed gases suitable for this purpose, as are e.g. oxygen, nitrogen and carbon dioxide. The pharmaceutically useful agents may also contain additives for conservation, stabilization, e.g. UV stabilizers, emulsifiers, sweetener, aromatizers, salts to change the osmotic pressure, buffers, coating additives and antioxidants. In general, in the case of oral or parenteral administration to adult humans weighing approximately 80 kg, a daily dosage of about 1 mg to about 10,000 mg, preferably from about 5 mg to about 1 ,000 mg, should be appropriate, although the upper limit may be exceeded when indicated. The daily dosage can be administered as a single dose or in divided doses, or for parenteral administration, it may be given as continuous infusion or subcutaneous injection.

EXAMPLES

Streptomyces albus carrying the xiamycin biosynthesis gene cluster was fermented in a large scale (50 L). Both mycelia and culture filtrate were extracted by ethyl acetate to afford a crude extract. After fractionation on flash chromatography on silica, followed by a Sephadex LH-20 column and final purification by HPLC, various dimers were obtained as pure substances. The NMR spectra of all dimers indicated that they have the xiamycin A backbone, yet according to HRESIMS data their molecular formula is C4 ₆ H ₄ 8N ₂ 0 ₆ (2a, 2b, 3a, 3b, and 4) and C ₄₆ H ₄₈ N ₂ S0 ₈ (5, 6 and 7), respectively. Consequently, they are dimers and sulfonyl dimers of xiamycin, respectively. After examination of NMR spectra respecially COSY and HMBC correlations, their structures were established. Figure 2 shows selected COSY and HMBC correlations for bixiamycins and sulfonylbixiamycins.

The Streptomyces albus strain has been deposited at the DSMZ (Inhoffenstr. 7B, 38124 Braunschweig, Germany) by the Leibniz Institute for Natural Product Research and Infection Biology e.V. - Hans-Knoll-Institute (HKI) (Beutenbergstr. 11a, 07745 Jena, Germany) on 20.08.2012 with the deposition number of DSM26342.

Compounds 2a/2b were a pair of atropoisomers. ¹³ C NMR spectra exhibited a highly conserved pattern as xiamycin. 1H NMR spectrum exhibited twelve aromatic protons. However, there were some differences in the chemical shifts for all protons in the Ή NMR spectrum. Based on the HRESIMS data, an only possibility is that 2a/2b were N-N coupled isomers. COSY and HMBC established the structure for the halve of the dimer. Compounds 3a/3b were another pair of atropoisomers. Their 1H NMR spectra showed two sets of signals, indicating that the two halves of the molecular were not identical. Only eleven aromatic protons were visible in the 1H NMR spectrum. As only a signal for H-21 was visible whereas the counterpart of Η-2 was missing in the 1H NMR spectrum, a N-C coupling between N-l and C-21 ' was proposed. COSY and HMBC established the structure for the two halves of the dimers.

Compound 4 was another C-N dimer. The 1H NMR spectra showed two sets of signals, indicating that the two halves of the molecular were not identical. Eleven aromatic signals were visible. COSY and HMBC spectra established the structures for both halves of the isomer. A N-C coupled isomer between N-2' and C-6 was proposed and fully established.

HRESIMS of Compound 5, 6 and 7 indicated the presence of a sulfonyl group. Compared to the above mentioned bixiamycins, there were dramatic changes in the UV spectra for 5, 6 and 7 compared to other bixiamycins, a substitution of sulfonyl group in the aromatic chromophore was proposed. Ή NMR spectrum of compound 5 exhibited eleven aomatic protons. Further analysis of COSY spectrum indicated a substitution at C-6. Thus, a sulfonyl group was located between N-2' and C-6. IR spectrum exhibited the characteristic absorption for sulfonyl group at 1244 and 1167 cm ^"1 , respectively.

Both 1H NMR and ¹³ C NMR spectra from compound 6 exhibited a similar pattern as xiamycin. However, only six aromatic protons (S 8.82, 7.97, 7.93, 7.53, 7.05) were observed. Based on the coupling constant and COSY spectra, the connection between H-5 (δ 8.82, d, 1.5 Hz), H-7 δ 7.97, dd, 8.6, 1.8 Hz) and H-8 δ 7.53, d, 8.6 Hz) were confirmed. Two other singlets (δ 7.93, 7.05) were from the carbazole ring H-10 and H-21, respectively. Based on the HRESIMS, a sulfonyl group between C(6)-C(6') was proposed. HMBC correlations fully confirmed the structure. IR spectrum exhibited the characteristic absorption for sulfonyl group at 1286 and 1126 cm ^"1 , respectively.

Compound 7 represents another sulfonyl bridged dimer of xiamycin as only ten aromatic protons were observed. Based on COSY and 1H NMR spectra, a sulfonyl function was located between C-6 and Η-2 and confirmed by HMBC correlations. IR spectrum exhibited the characteristic absorption for sulfonyl group at 1286 and 1121 cm ^"1 , respectively.

General Experimental Procedures. NMR spectra were recorded on a Bruker Avance III 500 MHz or Avance III 600 MHz spectrometers. IR spectra were recorded on a Bruker FT-IR (IFS 55) spectrometer. UV spectra were recorded on a Cary 1 Bio UV- visible spectrophotometer (Variant). Optical rotation was recorded on a Propol digital automatic polarimeter (Dr. Wolfgang Kernchen GmbH, Seelze, Germany). CD spectra were recorded on a J-810-150s spectropolarimeter (JASCO, GroB Umstadt, Germany). ESIMS data were obtained on a triple quadrupole mass spectrometer (Quattro; VG Biotech, Cheshire, UK). HRESIMS were recorded on a Finnigan TSQ Quantum Ultra AM Thermo Electron. Open column chromatography was performed on silica gel 60 (Merck, 0.04-0.063 mm, 230-400 mesh ASTM) and Sephadex LH-20 (Pharmacia). TLC analysis was performed on silica gel plates (Sil G/UV254, 0.20 mm, Macherey-Nagel). Preparative HPLC was performed on a Waters HPLC system using a Nucleosil 100-5 C ₁₈ column (5 μπι, 250 x 16 mm) with a UV detector.

Fermentation and Isolation. S. albus carrying the xia gene cluster was fermented in a 50 L scale fermentor for five days. The fermentation broth was separated into culture filtrate and mycelia by centrifugation. Both were extracted by ethyl acetate, evaporated into dryness and combined into a crude extract. Separation of the crude extract by flash silica gel chromatography (column 50 x 3 cm, CH ₂ C1 ₂ -CH ₃ 0H (0-100%) gradient, yielding ten fractions 1-10. Fraction 8 was fractionated by HPLC (RP-C ₁₈ , MeOH-H ₂ 0 as gradient) to yield four dimers: 2a (10 mg), 2b (5 mg), 3a (3 mg) and 3b (1.0 mg). Fraction 9 was fractionated by HPLC (RP-C ₁₈ , MeOH-H ₂ 0 as gradient) to yield three dimers: 4 (23.1 mg), 5 (0.5 mg) and 6 (0.8 mg). Fraction 10 containing 7 (1.5 mg) was purified to afford a pure substance.

Antimicrobial assays.

MIC assays were carried out to test all dimers and xiamycin against five bacterial test strains, Bacillus subtilis ATCC 6633, Staphylococcus aureus SG511, Mycobacterium vaccae IMET 10670, Staphylococus aureus 134/94, and Enterococcus faecalis 1528 R10. The assay was carried out by the standardized broth microdilution method (Clinical and Laboratory Standards Institute, 2006). Results are shown in Table 1.

Table 1. Antimicrobial activity of xiamcin, bixiamycins and sulfonylbixiamycins (MIC data, in μg ml/ ¹ ).

B. sub.: Bacillus subtilis ATCC 6633; S. aur. : Staphylococus aureus SG511; MRS A: Staphylococus aureus 134/94; VRE: vancomycin-resistant Enterococcus faecalis 1528 R10; M. vac: Mycobacterium vaccae IMET 10670; Cip: ciprofloxacin.

Antiproliferative and cytotoxic assays. All compounds were assayed against L-929 mouse fibroblasts (DSM ACC 2) and K562 human chronic myeloid leukemia cells (DSM ACC 10) for their antiproliferative effects (GI ₀ ), and against HeLa human cervix carcinoma cells (DSM ACC 57) for their cytotoxic (CC ₅₀ ) effects. Inhibitory concentrations are provided as 50% inhibition of cell growth (GI ₅₀ ; the concentration needed to reduce the growth of treated cells to half that of untreated cells) or 50% cytotoxic concentration (CC ₅₀ ; the concentration that kills 50% of treated cells). No activity was observed for all compounds (GI ₅₀ and CC ₅₀ >30 μg/mL). 1: pale yellow crystal; 137.6 (5.3 mg/mL, MeOH); UV/vis: ηαχ nm (log ε): 300 (4.02), 261 (4.05), 238 (4.34), 215 (4.23); CD (0.73 mg/mL, MeOH): Δε ₂₀₆ -9.56, Δε ₂₁₈ 8.47, Δε ₂₄₀ 21.74, Δε ₂₆₃ 8.17, Δε ₃₀₁ 4.83; IR(film): v _max : 3402.3, 3244.8, 2937.2, 1703.4, 1648.1, 1612.4, 1560.6, 1491.9, 1466.9, 1442.8, 1378.4, 1320.2, 1244.7, 1170.8, 1066.5, 1032.7, 1016.8, 940.8, 877.3, 767.8, 746.3, 717.7, 673.6, 666.5, 661.3, 652.2, 640.5 cm ^"1 ; ESIMS: [M - H] ^" = 362.2, [2M - H] ^" = 724.8, [M + H] ⁺ = 364.1, [M + NH ₄ ] ⁺ = 380.8, [2M + NH ₄ ] ⁺ = 743.9, [2M + Na] ⁺ = 749.0, HRESIMS: found [M - H] ^" 362.177 (calculated 362.1757 for C ₂₃ H ₂₄ N0 ₃ ).

2a: pale yellow solid; [a]" 42.8 (6.67 mg/mL, MeOH); UV/vis: η _¾χ nm (log ε): 333 (3.73), 319 (3.84), 296 (4.46), 250 (4.48), 233 (4.80), 217 (4.56); CD (0.11 mg/mL, MeOH): Δε ₂₀₃ - 4.00, Δε ₂₁₀ 3.90, Δε ₂ ι ₉ 3.58, Δε ₂₃₆ 10.46, Δε ₂₅₀ 2.92, Δε ₂₆₀ 2.87; IR (film) v _max 3399.9, 2935.1, 2875.3, 1703.8, 1627.6, 1609.3, 1468.5, 1455.9, 1232.3, 1067.4, 1019.2, 997.9 cm ^"1 ; ESIMS [M - H] ^" 723.3, [M + H] ⁺ 725.4; HRESIMS m/z 725.3602 [M + H] ⁺ (calcd. for C ₄₆ H ₄₉ N ₂ 0 ₆ 725.3585); NMR data, see Table 2.

2b: pale yellow solid; [c¾ ³ 82.9 (3.3 mg/mL, MeOH); UV/vis: ^nax nm (log ε): 332 (3.90), 319 (4.02), 296 (4.52), 250 (4.56), 234 (4.80), 217 (4.90); CD (0.11 mg/mL, MeOH): Δε ₂₀₇ - 9.67, Δε ₂₁₉ 9.00, Δε ₂₄₀ 19.73, Δε ₂₅₃ 8.57, Δε ₂₉₈ 11.65; IR (film) v _max 3337.2, 2944.7, 2833.9, 1652.7, 1449.2, 1416.5, 1 1 15.6, 1076.1, 1020.2 cm ^"1 ; ESIMS [M - H] ^" 723.3, [M + H] ⁺ 725.4, [M + NH ₄ ] ⁺ 742.4, [M + Na] ⁺ 747.3, NMR data, see Table 2.

Table 2. NMR data for compounds 2a and 2b (MeOD).

'H NMR 'H NMR ¹³ C NMR ^l3 C NMR

Position δ (Jin Hz) (600 MHz) δ (Jin Hz) (600 MHz) <5 (150 MHz) «5 (125 MHz)

2a 2b 2a 2b

1/Γ - - - -

2/2' - - 139.7 139.7

3/3' - - 122.1 121.4

4/4' - - 123.5 123.6

5/5' 8.01 (d, 7.7) 8.17 (d, 7.7) 121.8 121.8

6/6' 7.14 (t, 6.8) 7.27 (t, 6.8) 121.3 122.1 7/7' 7.12 (t, 6.8) 7.25 (t, 6.8) 127.2 127.2

8/8' 6.57 (d, 7.0) 6.73 (d, 6.8) 109.6 109.7

9/9' - - 144.4 144.5

10/10' 8.05 (s) 8.16 (s) 117.4 1 17.6

11/11 ' - - 141.4 141.6

12/12' - - 38.4 38.5

13/13' 2.54 (m); 1.71 (m) 2.69 (td, 13.1, 3.0); 1.80 (m) 38.8 38.9

14/14' 1.83 (m) 1.93 (m) 28.5 28.6

15/15' 4.04 (dd, 8.1 , 8.1) 4.09 (dd, 8.6, 7.1) 76.2 76.2

16/16' - - 54.7 54.8

17/17' 2.02 (dd, 12.5, 2.2) 2.13 (dd, 12.5, 2.0) 47.5 47.7

18/18' 1.72 (m); 1.30 (m) 1.92 (m); 1.45 (m) 22.2 22.3

19/19' 2.56 (m); 2.48 (m) 2.86 (m); 2.83 (m) 31.6 31.8

20/20' - - 135.7 135.8

21/21 ' 6.32 (s) 6.45 (s) 109.1 109.1

22/22' 1.16 (s) 1.33 (s) 26.3 26.2

23/23' 1.13 (s) 1.22 (s) 11.3 1 1.4

24/24' - 181.0 181.2

3a: pale yellow solid; [c¾ ³ 149.4 (0.55 mg/mL, MeOH); UV/vis: ^nax nm (log ε): 343 (3.83), 329 (3.87), 300 (4.38), 261 (4.39), 239 (4.69); CD (0.33 mg/mL, MeOH): Δε ₂₀₆ -29.07, Δε ₂₂₀ 12.27, Δε ₂₃₉ 24.50, Δε ₂₆₄ 16.72, Δε ₃₀₃ 22.67, Δε ₃₄₆ -5.97; IR (film) v _max 2930.3, 1715.4, 1698.0, 1457.0, 1236.2, 1063.6, 1018.2, 744.4 cm ^"1 ; ESIMS [M - H] ^" 723.3, NMR data, see Table 3.

Table 3. NMR data for 3a (MeOD).

'H NMR 'H NMR ¹³ C NMR ^,3 C NMR

Position δ (Jin Hz) (600 MHz) part δ (Jin Hz) (600 MHz) part <S (125 MHz) δ (125 MHz)

A B part A part B

2/2' - - 138.0 140.5

3/3' - - 124.7 123.4

4/4' - - 124.7 125.0 5/5' 8.05 (d, 7.8) 8.15 (d, 7.7) 120.8 121.1

6/6' 7.13 (t, 6.8) 7.19 (t, 7.0) 119.9 120.4

7/7' 7.26 (t, 6.8) 7.27 (t, 6.8) 126.6 126.6

8/8' 7.19 (d, 8.0) 6.79 (d, 8.0) 112.1 1 10.5

9/9' - - 142.1 142.2

10/10' 8.23 (s) 8.12 (s) 117.4 1 17.0

11/1 1 ' - - 143.0 143.1

12/12' - - 39.1 39.2

13/13' 2.69 (m); 1.77 (m) 2.74 (m); 1.95 (m) 38.4 38.6

14/14' 1.83 (m) 1.83 (m) 28.6 28.7

15/15' 4.07 (dd, 9.1 , 7.3) 4.11 (dd, 9.1 , 7.3) 76.3 76.2

16/16' - - 54.8 54.9

17/17' 2.12 (dd, 12.5, 2.1) 2.16 (dd, 12.5, 1.9) 47.6 47.9

18/18' 1.85 (m); 1.30 (m) 1.98 (m); 1.46 (m) 21.8 22.4

19/19' 2.42 (m); 2.37 (m) 2.94 (dd, 167, 6.4); 2.86 (m) 26.9 31.9

20/20' - - 133.2 135.0

21/21 ' - 6.53 (s) 1 18.7 109.7

22/22' 1.39 (s) 1.33 (s) 26.3 26.2

23/23' 1.23 (s) 1.19 (s) 1 1.3 11.4

24/24' _ _ 181.2 181.2

3b: pale yellow solid; [org 25.2 (0.55 mg/mL, MeOH); UV/vis: nm (log ε): 343 (3.83), 329 (3.85), 300 (4.38), 261 (4.38), 239 (4.68), 218 (4.51); CD (0.73 mg/mL, MeOH): Δε ₂₁₂ 7.48, Δ¾ ₂₀ -1.33, Δε ₂₃₇ 18.21, Δε ₂₅₂ 4.86, Δε ₂₆₄ -5.32, Δε ₂₈₈ 1.42, Δε ₃₀₄ -6.56; IR (film) v _max 2925.5, 2853.2, 1464.7, 1068.4, 1018.2, 999.0, 743.4 cm ; ESIMS [M - H] ^" 723.5, NMR data, see Table 4.

Table 4. NMR data for 3b (MeOD).

'H NMR 'H NMR ¹³ C NMR ¹³ C NMR

Position S (J in Hz) (500 MHz) part δ (J in Hz) (500 MHz) part δ (150 MHz) <5 (150 MHz)

A B part A part B

1/Γ

2/2' 138.0 140.4 3/3' - - 124.7 123.3

4/4' - - 124.7 125.0

5/5' 8.09 (d, 7.8) 8.15 (d, 7.8) 120.8 121.0

6/6' 7.13 (t, 7.9) 7.20 (t, 7.0) 119.9 120.5

7/7' 7.26 (t, 6.8) 7.27 (t, 6.8) 126.6 126.7

8/8' 7.20 (d, 7.7) 6.80 (d, 8.1) 112.0 110.3

9/9' - - 142.1 142.3

10/10' 8.23 (s) 8.12 (s) 117.5 117.1

11/11 ' - - 143.2 142.9

12/12' - - 38.6 38.4

13/13' 2.76 (m); 1.95 (m) 2.69 (m); 1.85 (m) 39.0 39.2

14/14' 1.93 (m) 1.93 (m) 28.6 28.6

15/15' 4.11 (dd, 9.1, 7.5) 4.11 (dd, 9.1, 7.5) 76.3 76.2

16/16' - - 54.9 54.8

17/17' 2.16 (d, 12.5) 2.16 (d, 12.5) 47.5 47.8

18/18' 1.83 (m); 1.27 (m) 1.93 (m); 1.48 (m) 22.5 21.8

19/19' 2.37 (m) 2.92 (m) 26.7 32.0

20/20' - - 133.3 135.0

21/21 ' - 6.51 (s) 118.7 109.7

22/22' 1.40 (s) 1.31 (s) 26.4 26.3

23/23' 1.22 (s) 1.19 (s) 11.4 11.4

24/24' - - 181.5 181.5

4: pale yellow solid; [af* 133.9 (1.63 mg/mL, MeOH); UV/vis: Λ™ _χ nm (log ε)\ 333 (3.78),

301 (4.33), 265(4.39), 240 (4.69), 219 (4.42); CD (0.38 mg/mL, MeOH): Δε ₂₀₆ -9.70, Δε ₂₂₅ 9.64, Δε ₂₄₂ 19.74, Δε ₂₆₆ 8.41, Δε ₃₀₃ 4.68, Δε ₃₅₃ -2.86; IR (film) v _max 2938.0, 2833.8, 1697.1, 1494.6, 1468.5, 1235.2, 1018.2, 745.4 cm ^"1 ; ESIMS [M - H] ^" 723.5, NMR data, see Table 5.

Table 5. NMR data for 4 (MeOD).

'H NMR 'H NMR ¹³ C NMR ^I3 C NMR

Position δ (Jin Hz) (600 MHz) part δ (J in Hz) (600 MHz) part «5 (125 MHz) <5 (125 MHz)

A B part A part B 2/2' - - 140.9 142.0

3/3' - - 122.7 122.9

4/4' - - 125.4 124.5

5/5' 7.97 (d, 2.0) 7.99 (d, 8.2) 120.2 120.7

6/6' - 7.08 (t, 7.8) 129.8 120.2

7/7' 7.21 (dd, 8.3, 2.0) 7.16 (t, 7.3) 125.6 126.4

8/8' 7.39 (d, 8.4) 7.1 1 (d, 8.0) 1 12.4 1 10.5

9/9' - - 140.7 143.7

10/10' 7.81 (s) 7.96 (s) 1 16.7 1 16.5

11/11 ' - - 142.2 142.5

12/12' - - 38.2 38.3

13/13' 2.29 (d, 11.4); 1.45 (m) 2.56 (d, 13.1); 1.65 (m) 38.7 38.9

14/14' 1.62 (m) 1.83 (m) 28.6 28.4

15/15' 3.96 (dd, 10.9, 4.8) 4.05 (dd, 8.2, 7.9) 76.2 76.2

16/16' - - 54.7 54.8

17/17' 2.06 (t, 14.2) 2.06 (t, 14.2) 47.6 47.7

18/18' 1.95 (m); 1.52 (m) 1.77 (m); 1.34(m) 22.4 22.4

19/19' 2.98 (m) 2.74 (m) 31.9 31.9

20/20' - - 134.9 134.5

21/21 ' 6.99 (s) 6.85 (s) 111.2 109.9

22/22' 1.08 (s) 1.18 (s) 26.2 26.3

23/23' 1.17 (s) 1.14 (s) 1 1.3 11.4

24/24' _ _ 181.2 181.3

5: pale yellow solid; [a * 134.1 (0.50 mg/mL, MeOH); UV/vis: ^nax nm (log ε): 285 (4.46), 224 (4.58); CD (0.33 mg/mL, MeOH): Δε ₂₃₀ 19.12, Δε ₂₉₀ 9.50; IR (film) v _max 2927.4, 2853.2, 1652.7, 1243.9, 1 166.7, 1 136.8, 1016.3, 937.2 cm ^"1 ; HRESIMS m/z 789.3217 [M + H] ⁺ , (calcd. for C ₄₆ H ₄₉ N ₂ S0 ₈ 789.3204); NMR data, see Table 6.

Table 6. NMR data for 5 (MeOD).

, _{H NMR} 'H NMR ^I3 C NMR ¹³ C NMR

Position δ (J in Hz) (500 MHz) part δ (Jin Hz) (500 MHz) part δ (150 MHz) <5 (150 MHz)

A B part A part B ι/r

2i r - - 140.7 138.3

3/3' - - 122.2 126.2

4/4' - - 124.1 128.2

5/5' 8.56 (s) 7.89 (d, 8.6) 120.5 120.8

6/6' - 7.29 (t, 7.4) 127.8 124.9

7/7' 7.65 (d, 8.8) 7.45 (t, 7.4) 124.1 127.8

8/8' 7.89 (d, 7.6) 8.35 (d, 8.4) 111.7 116.3

9/9' - - 144.7 140.1

10/10' 7.82 (s) 7.82 (s) 116.8 116.9

11/11 ' - - 143.6 146.9

12/12' - - 38.3 38.3

13/13' 2.49 (m); 1.63 (m) 2.32 (m); 1.53 (m) 38.4 38.4

14/14' 1.82 (m) 1.75 (m) 28.4 28.4

15/15' 4.03 (m) 4.03 (m) 76.2 76.2

16/16' - - 54.8 54.8

17/17' 2.08 (m) 2.08 (m) 47.5 47.4

18/18' 1.92 (m); 1.53 (m) 1.92 (m); 1.53 (m) 22.4 22.4

19/19' 2.85 (m) 3.15 (m); 3.05 (m) 31.9 32.3

20/20' - - 136.2 136.3

21/21 ' 7.03 (s) 8.08 (s) 111.5 116.0

22/22' 1.14 (s) 1.14 (s) 26.0 26.0

23/23' 1.19 (s) 1.19 (s) 11.5 11.5

24/24' 181.7 181.7

6: pale yellow solid; [a] ² 65.2 (0.90 mg/mL, MeOH); UV/vis: η _3Χ nm (log ε): 293 (4.38), 272 (4.31), 220 (4.37); CD (0.68 mg/mL, MeOH): Δ¾ ₃₆ 18.95, Δε ₂₇₅ 8.25; IR (film) v _max 3335.3, 2931.3, 1602.6, 1286.3, 1242.9, 1141.7, 1126.2, 875.5, 804.2 cm ⁴ ; ESIMS [M - H] ^" 787.3, NMR data, see Table 7.

Table 7. NMR data for 6 (MeOD).

! _{H NMR} ¹³ C NMR

Position

δ (J in Hz) (600 MHz) δ (150 MHz) 1/Γ

2/2' - 140.8

3/3' - 122.5

4/4' - 124.8

5/5' 8.82 (d, 1.5) 120.8

6/6' - 133.5

7/7' 7.97 (dd, 8.6, 1.8) 125.2

8/8' 7.47 (d, 8.6) 112.0

9/9' - 144.2

10/10' 7.93 (s) 117.1

1 1/11 ' - 143.4

12/12' - 38.3

13/13' 2.36 (m); 1.54 (m) 38.6

14/14' 1.82 (m) 28.5

15/15' 4.01 (dd, 11.4, 5.0) 76.3

16/16' - 54.8

17/17' 2.05 (dd, 12.5, 2.0) 47.6

18/18' 1.93 (m); 1.52(m) 22.4

19/19' 3.04 (dd, 16.7, 6.6); 2.96 (m) 31.9

20/20' - 135.7

21/21 ' 7.05 (s) 1 1 1.4

22/22' 1.07 (s) 26.0

23/23' 1.19 (s) 1 1.4

24/24' _ 181.6

7: pale yellow solid; [af 68.0 (0.80 mg/mL, MeOH); UV/vis: Anax nm (log έ): 360 (3.94), 346 (3.95), 290 (4.58), 222 (4.76); CD (0.50 mg/mL, MeOH): Δ¾π 6.73, Δ¾ ₃₈ 18.21 , Δε ₂₇ ι 9.97, Δε ₂₈₉ -7.12, Δε ₃₁₁ 2.36; IR (film) v _raax 2931.3, 2840.6, 1652.7, 1467.6, 1286.3, 1241.0, 1 121.4, 1015.3 cm ^"1 ; ESIMS [M - H] ^" 787.3, NMR data, see Table 8.

Table 8. NMR data for 7 (MeOD).

'H MR 'H NMR ¹³ C NMR ¹³ C NMR

Position

δ (Jin Hz) (600 MHz) δ (Jin Hz) (600 MHz) δ (150 MHz) δ (150 MHz) part A part B part A part B

2/2' - - 140.9 138.2

3/3' - - 122.4 125.7

4/4' - - 124.3 123.2

5/5' 8.61 (d, 1.7) 8.12 (d, 8.9) 120.5 120.9

6/6' - 7.24 (dd, 7.8, 7.3) 132.9 120.8

7/7' 7.75 (dd, 8.7, 1.7) 7.45 (t, 7.3) 124.7 127.7

8/8' 7.43 (d, 8.7) 7.65 (d, 8.1) 111.8 112.5

9/9' - - 144.4 141.7

10/10' 8.00 (s) 8.42 (s) 117.2 123.4

11/11 ' - - 143.6 143.0

12/12' - - 38.8 39.2

13/13' 2.57 (m); 1.67 (m) 2.61 (m); 1.67 (m) 38.4 39.0

14/14' 1.84 (m) 1.84 (m) 28.5 28.5

15/15' 4.03 (dd, 8.9, 8.5) 4.05 (dd, 9.1, 7.3) 76.3 75.9

16/16' - - 54.7 54.6

17/17' 2.12 (dd, 11.5, 2.0) 1.92 (dd, 12.6, 1.5) 47.7 46.6

18/18' 2.00 (m); 1.55 (m) 1.79 (m); 1.45 (m) 22.4 21.8

19/19' 3.11 (dd, 6.6, 6.4); 3.03 (m) 3.49 (dd, 8.2, 7.9); 2.89 (m) 32.0 29.2

20/20' - - 136.1 134.5

21/21 ' 7.13(s) - 111.6 121.4

22/22' 1.25 (s) 1.24 (s) 26.0 26.2

23/23' 1.21 (s) 1.13 (s) 11.5 11.4

24/24' - - 181.6 181.6

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