EA039955B1 - MACROCYCLIC DEAZA-OXYPURINES AND THEIR USE FOR THE TREATMENT OF VIRAL INFECTIONS - Google Patents

Description

The present invention relates to macrocyclic deaza-oxypurine derivatives, pharmaceutical compositions and their use in the treatment of viral infections.

The present invention relates to the use of macrocyclic deazaoxypurine derivatives in the treatment of viral infections, immune or inflammatory disorders involving the modulation or agonism of toll-like receptors 7 (TLR7). Toll-like receptors are major transmembrane proteins characterized by an extracellular leucine-rich domain and a cytoplasmic extension that contains a conserved region. The innate immune system can recognize pathogen-associated molecular patterns through TLR data expressed on the cell surface of certain types of immune cells. When foreign pathogens are recognized, the production of cytokines is activated and the expression of costimulatory molecules on phagocytes increases. This leads to modulation of T cell behavior.

It has been established that most mammalian species have ten to fifteen types of toll-like receptors. A total of thirteen TLRs (named TLR1-TLR13) have been identified in humans and mice, and equivalent forms of many have been found in other mammalian species. However, equivalents of certain TLRs found in humans are not present in all mammals. For example, a gene encoding a protein similar to TLR10 in humans is present in mice but appears to have been damaged by a retrovirus at some point in the past. On the other hand, mice express TLRs 11, 12 and 13, none of which are present in humans. Other mammals may express TLRs that are not found in humans. Other non-mammalian species may have different TLRs than mammals, as evidenced by TLR14 found in puffer fish of the genus Takifugu. This may complicate the procedure for using experimental animals as models of human innate immunity.

For a detailed review of toll-like receptors, see the following journal articles. Hoffmann, J.A., Nature, 426, p33-38, 2003; Akira, S., Takeda, K., and Kaisho, T., Annual Rev. Immunology, 21, p335-376, 2003; Ulevitch, R. J., Nature Reviews: Immunology, 4, p512-520, 2004.

Compounds having activity at toll-like receptors have previously been described, such as purine derivatives in WO 2006/117670, adenine derivatives in WO 98/01448 and WO 99/28321, and pyrimidines in WO 2009/067081.

However, there is a strong need for new toll-like receptor modulators having advantageous selectivity, higher potency, higher metabolic stability, and an improved safety profile compared to prior art compounds.

In the treatment of certain viral infections, regular injections of interferon (IFN-alpha) may be given, as is the case with hepatitis C virus (HCV). For more information, see Fried et. al. Peginterferon-alfa plus ribavirin for chronic hepatitis C virus infection, N Engl J Med 2002; 347:975-82. Small molecule IFN inducers available for oral administration offer the potential benefits of reduced immunogenicity and ease of administration. Thus, new IFN inducers represent a potentially effective new class of drugs for the treatment of viral infections. For an example of a small molecular weight IFN inducer with an antiviral effect, see De Clercq, E.; Descamps, J.; De Somer, P. Science 1978, 200, 563-565.

IFN-alpha is also administered in combination with other drugs in the treatment of certain types of cancer (Eur. J. Cancer 46, 2849-57, and Cancer Res. 1992, 52, 1056). TLR 7/8 agonists are also of interest as vaccine adjuvants due to their ability to induce a pronounced Th1 response (Hum. Vaccines 2010, 6, 322-335, and Hum. Vaccines 2009, 5, 381-394).

1) According to the present invention, there is provided a compound selected from the group consisting of:

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hh ₂ NH ₂ N ^ X _N 'TG ^ nh ₂ ^HO ^ XX w 1 o o o X o O O Λ _ζ δ zA ² A _z A _{z Z} A _Z ZI Ο ZI Ο Ή /° °\ Ή )° ° /° θ P °\ W w H SA nh ₂ n^X _n ^h °XI if L ⁿ /X u nh ₂ a ^N ^N noD 1 A v °\ nh ₂ Ν>Λ ho—PL if XX nh ₂ noD Y l Xr nh ₂ ^h °—LX Yx : ¹ nh ₂ ^Η0 ^ ^/Ν Χχ V ⁵ F nh ₂ <Q°X> nh ₂ ^HO X ^/N Xx XX nh ₂ ⁿ °^xY U _F nh ₂ N-^Xn ^H °x ^z Xjl , HCI [I nh ₂ ^Η °X ^/Ν Χχ N and nh ₂ .ns ^k Q°^ F nh ₂ but-^ D /1 nh ₂ ^HO \ X F nh ₂ HQ—c L if ^{hci ι} χχ ^ο< ^ F nh ₂ ^but -/ X j F nh ₂ ^HO ^\ X v ^A F nh _2HO ^XX JL νη ₂ noD LX nh ₂ N^'N ho—L if

and its pharmaceutically acceptable salts.

Also part of the present invention is a pharmaceutical composition comprising said compounds or a pharmaceutically acceptable salt thereof together with one or more pharmaceutically acceptable excipients, diluents or carriers.

In addition, the present invention also relates to the above compound or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition for use as a medicine.

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The present invention also relates to the above compound, or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition, for use in the treatment of a disorder in which TLR7 modulation is involved.

Pharmaceutically acceptable salts of the compounds of formula (I) include their acid addition salts and base addition salts. Suitable acid addition salts are formed from acids which form non-toxic salts. Suitable base salts are formed from bases which form non-toxic salts.

The compounds of the present invention may be present in the form of so-called tautomer(s), which refers to isomers of organic compounds that readily undergo interconversion by a chemical reaction called tautomerization. The result of this reaction is the symmetrical migration of a hydrogen atom or proton, accompanied by a shift of the single bond and the adjacent double bond.

The compounds of the present invention may be administered as crystalline or amorphous products. They can be obtained, for example, in the form of a solid compact mass, powders or films by methods such as precipitation, crystallization, freeze drying, spray drying or evaporation drying. They can be administered alone or in combination with one or more other compounds according to the present invention, or in combination with one or more other drugs. Typically, they will be administered as a formulation in combination with one or more pharmaceutically acceptable excipients. The term excipient is used herein to describe any ingredient other than the compound(s) of the present invention. The choice of excipient largely depends on such factors as the particular route of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

The compounds of the present invention, or any subgroup thereof, may be formulated into various pharmaceutical forms for purposes of administration. As suitable compositions there may be mentioned all compositions commonly used for the systemic administration of drugs. To prepare pharmaceutical compositions according to the present invention, an effective amount of a particular compound, optionally in the form of an addition salt, as an active ingredient is combined in a homogeneous mixture with a pharmaceutically acceptable carrier, the carrier may take a wide variety of forms depending on the form of preparation required for administration. . These pharmaceutical compositions are preferably presented in unit dosage form suitable, for example, for oral, rectal or transdermal administration. For example, when preparing compositions in the form of an oral dosage form, any conventional pharmaceutical medium can be used, such as, for example, water, glycols, oils, alcohols, and the like, in the case of oral liquid preparations, such as suspensions, syrups, elixirs, emulsions. and solutions; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrants and the like, in the case of powders, pills, capsules and tablets. Because of their ease of administration, tablets and capsules are the most preferred oral unit dosage forms, in which case, of course, solid pharmaceutical carriers are used. Also included are solid form preparations which can be converted shortly before use to liquid form preparations. In compositions suitable for transdermal administration, the carrier optionally comprises a penetrating agent and/or a suitable wetting agent, optionally in combination with suitable additives of any nature in minimal proportions, without the additives having a significant deleterious effect on the skin. Said additives may facilitate administration to the skin and/or may be useful in preparing the desired compositions. These compositions can be administered in various ways, for example in the form of a transdermal patch, in the form of a spot application, in the form of an ointment. Compounds in accordance with the present invention can also be administered by inhalation or insufflation using methods and formulations used in the art for administration by this route. Thus, in general, the compounds of the present invention can be administered to the lungs in the form of a solution, suspension or dry powder.

It is particularly preferred to formulate the above pharmaceutical compositions in unit dosage form for ease of administration and uniform dosing. The unit dosage form used herein refers to physically separate units suitable as unit doses, each unit containing a predetermined amount of the active ingredient, calculated to produce the desired therapeutic effect, in combination with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including divisible or coated tablets), capsules, pills, powder sachets, cachets, suppositories, injectable solutions or suspensions, and the like, as well as their separate sets.

Those skilled in the art of treating infectious diseases will be able to determine the effective amount based on the results of the tests presented later in this document. In general, they believe that

- 3 039955 an effective daily amount will be from 0.01 to 50 mg/kg of body weight, more preferably from 0.1 to 10 mg/kg of body weight. It may be appropriate to administer the required dose in two, three, four or more dose portions at appropriate intervals throughout the day. Said dose portions may be formulated as unit dosage forms, for example containing from 1 to 1000 mg and in particular from 5 to 200 mg of the active ingredient per dosage unit.

The exact dosage and frequency of administration will depend upon the particular compound used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient, as well as other medical treatments the individual may be receiving, as is well known to those skilled in the art. areas. Moreover, it is obvious that the effective amount can be reduced or increased depending on the response of the subject being treated and/or depending on the judgment of the physician prescribing the compounds in accordance with the present invention. Thus, the above effective amount ranges are only guidelines and are not intended to limit in any way the scope or application of the present invention.

General scheme for obtaining final products: method 1.

F1

G1 B1

Synthesis of intermediate B1.

SOCl ₂ (80 ml; 1.11 mol) was added dropwise to a mixture of G1 (19.1 g; 111 mmol) in CH ₂ Cl ₂ (230 ml) at rt. The mixture was stirred at rt. within 16 hours the Solvent was evaporated, and the residue was solubilized in CH ₂ Cl ₂ and treated with saturated aqueous NaHCO ₃ to basic pH. The layers were separated and the aqueous layer was extracted with CH ₂ Cl ₂ (twice). The combined organic layers were dried over MgSO4, filtered and concentrated in vacuo to give 20.1 g of a brown oil. The crude compound was used in the next step without further purification.

C1

Synthesis of intermediate C1.

The reaction was carried out in parallel on two batches (13.6 mmol and 27.1 mmol Al).

To a solution of A1 (5.0 g; 13.6 mmol) and K ₂ CO ₃ (3.75 g; 27.1 mmol) in acetone (100 ml) was added B1 (4.46 g; 24.4 mmol) and NaI (2.23 g; 14.9 mmol) at RT The mixture was stirred at rt. for 16 hours. The mixture was filtered and the filtrate was evaporated in vacuo to give a brown oil. Two batches were combined and purified by preparative LC (irregular SiOH 15-40 µm, 220 g Grace, mobile phase, gradient: CH ₂ Cl ₂ /EtOAc 100/0 to 80/20). Fractions containing the product were combined and the solvent was removed in vacuo to give 18.6 g of intermediate C1 (89% yield).

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Synthesis of intermediate D1.

To a solution of C1 (18.6 g; 36.1 mmol) in THF (300 ml) was added an aqueous solution of NH ₃ (30%) (290 ml) at rt. and the mixture was stirred at rt. for 16 hours. The mixture was absorbed with EtOAc and saturated NaCl solution, the layers were separated and the organic layer was dried over MgSO 4 , filtered and the solvent was removed under reduced pressure to give 16.7 g of a yellow-orange oil. The crude product was dried under high vacuum to give 16.5 g of a sticky yellow-orange solid which was used directly in the next step.

Synthesis of intermediate compound E1.

NaH (60% in oil) (1.75 g; 43.7 mmol) was added in portions to allyl alcohol (50 ml) at rt. The mixture was stirred at rt. for 30 min before adding dropwise to a solution of D1 (5 g; 11.1 mmol) in THF (124 ml) at 0°C. The resulting mixture was then stirred at rt. for 1 h and poured into a saturated aqueous solution of NH ₄ Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated and the aqueous layer was extracted with EtOAc (once). The combined organic layers were dried over MgSO ₄ , filtered and the solvent removed under reduced pressure to give a yellow oil. The crude compound was purified by preparative LC (15-40 μm irregular SiOH, 120 g Grace, liquid injection, mobile phase, gradient: heptane/EtOAc 100/0 to 50/50) to give 4.04 g of Intermediate E1 in as a yellow oil (79% yield).

Synthesis of intermediate compound F1.

The reaction was performed in 2 batches of 850 mg and 2 batches of 1 g of E1.

The procedure for one batch of 850 is given.

In a Schlenk flask, a solution of E1 (0.85 g; 1.98 mmol) and chlordicyclohexylborane (1M solution in hexane) (400 µl; 400 µmol) in dichloroethane (570 ml) were stirred at 80°C under N ₂ atmosphere for 1 h Grubbs-Hoveyda 2nd generation catalyst (124 mg; 198 µmol) was added and the mixture was stirred at ₁₂₀ ° C. for 16 h. generation (124 mg; 198 µmol) and chlordicyclohexylborane (1M solution in hexane) (400 µl; 400 µmol). The mixture was stirred at 120°C for 20 h.

2 batches were mixed and ruthenium scavenger (DMT SiliaBond from SiliCycle) (10.4 g; 6.35 mmol) was added and the mixture was stirred at rt. for 20 hours. The reaction mixture was filtered through a celite pad and the solvent was removed under reduced pressure to give a brown residue.

The residue was combined with the residue from two batches of 1 g E1. The resulting brown residue was purified by preparative LC (SiOH 15-40 µm irregular grains, 120 g Grace, dry loading, mobile phase, gradient: heptane/EtOAc 100/0 to 0/100) to give 1.19 g of a brown solid substances. The brown solid was further purified by preparative LC (stationary phase: open surface irregular silica gel 40 g, mobile phase gradient: CH ₂ Cl ₂ /EtOAc 90/10 to 80/20) to give 705 mg of a yellow solid. The yellow solid was further purified with achiral SFC (stationary phase: Amino 6 μm 150x21.2 mm), mobile phase: gradient from 85% CO ₂ , 15% MeOH to 65% CO ₂ , 35% MeOH) to give 660 mg of Intermediate F1 as a yellow solid (19% yield, E-isomer).

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Synthesis of final compound 1.

A mixture of F1 (570 mg; 1.42 mmol) and iron (795 mg; 14.2 mmol) in AcOH (21 ml) and water (4.2 ml) was stirred at 50° C. for 2 hours. The mixture was concentrated to dryness. DMF was added, the mixture was sonicated, heated and filtered through a Celite pad, and the Celite was washed with hot DMF. Iron scavenger (SiliaBond imidazole from SiliCycle) (25.4 g; 29.5 mmol) was added to the filtrate and the mixture was stirred at rt. for 16 hours. The mixture was filtered through celite, the celite was washed with DMF and the filtrate was concentrated in vacuo to give 620 mg of a brown solid. The crude material was purified by preparative LC (irregular SiOH, 15-40 µm, 30 g Merck, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq. 98/2/0.2 to 85/15/ 1.5) to give 360 mg of final compound 1 as an off-white solid (75% yield).

C1

Alternative Synthesis of Intermediate C1 At 0°C, diisopropyl azodicarboxylate (DIAD) (3.0 ml, 15.0 mmol) was added dropwise to a mixture of A1 (3.70 g, 10.028 mmol), G1 (1.98 g, 12.0 mmol ) and PPh ₃ (3.94 g, 15.0 mmol) in THF (70 ml). The mixture was stirred at rt. within 12 hours Added EtOAc and water. The layers were decanted. The organic layer was washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated. The crude material was purified by preparative LC (irregular SiOH 20-45 µm 450 g Matrex), mobile phase (85% heptane, 15% AcOEt) to give 4.5 g of intermediate C1 (87% yield).

General scheme for obtaining final products: method 2.

F1 H1

Synthesis of intermediate H1.

Wilkinson's catalyst (44 mg; 47.5 µmol) was added to a solution of F1 (190 mg; 475 µmol) in THF/MeOH (50/50) (50 mL) purged with N ₂ sparge for 15 min. The mixture was hydrogenated (8 bar) at rt. for 16 hours. The mixture was purged for 15 minutes and additional Wilkinson's catalyst (44 mg; 47.5 µmol) was added. The reaction mixture was hydrogenated (8 bar) at rt. for 4 h. The mixture was concentrated in vacuo to give a brown oil. The oil was purified by preparative LC (irregular SiOH 15-40 µm, 12 g Grace, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /EtOAc 100/0 to 80/20) to give 150 mg of Intermediate H1 as a yellow solid (79% yield).

Synthesis of final compound 2.

Compound 2 was obtained using the procedure for obtaining compound 1 (54 mg, 44% yield).

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General scheme for obtaining final products: method 3.

Vg

Graobs Hoveida

Synthesis of intermediate J1.

I1 (5.9 g; 35.6 mmol) was added to a solution of A1 (7.3 g; 19.8 mmol), K ₂ CO ₃ (5.5 g; 39.6 mmol) and NaI (3.3 g ; 21.8 mmol) in acetone (145 ml). The mixture was stirred at rt. for 20 hours. The mixture was filtered through a Celite pad and the filtrate was evaporated in vacuo to give an orange solid. The residue was taken up in CH2Cl2. The precipitate was filtered and the filtrate was concentrated in vacuo to give 13 g of a yellow oil.

The crude compound was purified by preparative LC (irregular SiOH 15-40 µm, 300 g Interchim, mobile phase, gradient: heptane/EtOAc 100/0 to 80/20). Fractions containing product were combined and the solvent was removed in vacuo to give 7.1 g of Intermediate J1 (72% yield) as a yellow oil.

Synthesis of intermediate compound K1.

In a Schlenk flask, a solution of J1 (7.1 g; 14.2 mmol) in THF (130 ml) and an aqueous solution of NH ₃ (30%) (130 ml) were stirred at rt. within 16 hours the Mixture was absorbed with EtOAc and saturated aqueous NaCl solution, the layers were separated. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo to give 6.4 g of a yellow oil (100% yield).

Synthesis of intermediate compound L1.

NaH (2.2 g; 54.2 mmol) was added in portions at rt. and under N ₂ to 3-buten-1-ol (76 ml). The mixture was stirred at rt. for 30 minutes before adding it dropwise at 0° C. to a solution of K1 (5.9 g; 13.6 mmol) in THF (150 ml). The resulting mixture was stirred at 0° C. for 1 h. The mixture was poured into an aqueous saturated solution of NH ₄ Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated. The organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo to give a yellow residue which was azeotropically distilled off with toluene (once) to give 6.6 g of a yellow oil. The crude compound was purified by preparative LC (irregular SiOH 15-40 µm, 220 g Grace, mobile phase, gradient: heptane/EtOAc 100/0 to 50/50). Fractions containing

The product was combined and the solvent was removed in vacuo to give 4.46 g of Intermediate L1 (77% yield) as a yellow oil.

Synthesis of the intermediate compound Ml.

The reaction was performed in 2 batches.

Typical procedure for one batch.

A solution of L1 (2.45 g; 5.75 mmol) in dry CH ₂ Cl ₂ (1.7 L) was degassed by bubbling N ₂ for 15 minutes. Grubbs 2nd generation catalyst (488 mg; 574 µmol) was added and the mixture was stirred at rt. over 72 h. DMT SiliaBond (7.66 g; 4.59 mmol) was added and the mixture was stirred at rt. within 16 hours Combined 2 batches and filtered through celite. The filtrate was concentrated in vacuo to give a black solid. The crude compound was purified by preparative LC (irregular SiOH 15-40 µm, 150 g Merck, mobile phase, gradient: heptane/EtOAc 100/0 to 50/50). Fractions containing the product were combined and the solvent was removed in vacuo to give 230 mg of fraction 1 and 2.3 g of fraction 2. Fraction 2 was previously purified by preparative LC (stationary phase: SiOH with irregular grains of 40 μm 120 g, mobile phase: heptane/ CH ₂ Cl ₂ /MeOH 55/43/2). The isolated compound was combined with fraction 1 and purified with achiral SFC (stationary phase: Chiralpak IC 5 μm 250x20 mm, mobile phase: 70% CO ₂ , 30% iPrOH) to give 1.51 g of intermediate M1 (33% yield, isomer E) as a yellow solid.

Synthesis of final compound 3.

Iron (631 mg; 11.3 mmol) was added to a solution of M1 (750 mg; 1.88 mmol) in AcOH (150 ml) and water (25 ml). The mixture was stirred at 80°C for 16 hours. Iron (315 mg; 5.65 mmol) was added and the mixture was stirred at 80°C for 2 hours. Iron (315 mg; 5.65 mmol) was added and the mixture was stirred at 80 °C for 4 hours. Iron (315 mg; 5.65 mmol) was added and the mixture was stirred at 80°C for 16 hours. The mixture was concentrated to dryness. DMF was added, the mixture was filtered through Celite and the Celite was washed with hot DMF. SiliaBond imidazole (48.7 g; 56.5 mmol) was added to the filtrate and the mixture was stirred at rt. for 16 hours. The mixture was filtered through celite, the celite was washed with DMF and the filtrate was concentrated in vacuo. The crude compound was purified by preparative LC (irregular SiOH, 15-40 µm, 25 g Merck, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq 98/2/0.2 to 85/15 /1.5) to obtain 2 fractions. Fraction 1 was absorbed with EtOH and filtered to give fraction 3 and fraction 2 was absorbed with MeCN and filtered to give fraction 4. Fractions 3 and 4 were combined in EtOH, filtered and dried in vacuo to give 199 mg of final compound 3 (33% exit).

General scheme for obtaining final products: method 4.

Synthesis of intermediate N1.

Wilkinson's catalyst (46 mg; 50.2 µmol) was added to a solution of M1 (200 mg; 502 µmol) in THF/MeOH (50/50) (50 ml) purged by bubbling N ₂ for 15 min. The mixture was hydrogenated (7 bar) at rt. for 20 h. The mixture was purged by bubbling N ₂ for 15 min, additional Wilkinson's catalyst (46 mg; 50.2 µmol) was added and the reaction mixture was hydrogenated (7 bar) at rt. within 16 hours the Reaction mixture was concentrated in vacuo to obtain a green oil. The oil was purified by preparative LC (SiOH 15-40 μm irregular grains, 25 g Merck, dry loading, mobile phase, gradient: heptane/EtOAc 100/0 to 70/30) to give 130 mg of intermediate N1 as yellow solids (66% yield).

Synthesis of intermediate compound 01.

In a high pressure reactor, N1 (110 mg; 275 µmol) was hydrogenated in EtOH (5 ml) with Pd/C

- 8 039955 (10%) (30 mg; 28.5 µmol) as catalyst at 40°C (3 bar) for 6 hours. The catalyst was removed by filtration through celite, the celite was washed with EtOH and the filtrate was evaporated under vacuum with obtaining 100 mg of a yellow residue (98% yield). Intermediate 01 was used in the next step without further purification.

Synthesis of final compound 4.

In a sealed test tube 01 (100 mg; 270 µmol) in pure acetic acid (5 ml) was stirred at room temperature. within 90 min. The solvent was removed under reduced pressure to give a yellow residue. The residue was absorbed with CH ₂ Cl ₂ and the solvent was removed under reduced pressure (twice) to give 87 mg of a yellow-green solid. The solid was azeotropically distilled off with toluene (four times) and then triturated and sonicated in Et2O. The mixture was filtered (glass frit n°5) to give 75 mg of final compound 4 (77% yield, acetate salt).

General scheme for obtaining final products: method 5.

Q1

Synthesis of intermediate compound Q1.

To a solution of A1 (3.52 g; 9.54 mmol) and K ₂ CO ₃ (2.64 g; 19.1 mmol) in acetone (80 ml) was added P1 (1.93 g; 10.5 mmol) and NaI (1.57 g; 10.5 mmol) at RT The mixture was stirred at rt. for 16 h, then P1 (1.5 g; 8.17 mmol) was added and the mixture was stirred at rt. for 24 hours. The reaction mixture was filtered through a Celite pad and the filtrate was evaporated in vacuo to give a black residue. The residue was purified by preparative LC (irregular SiOH 15-40 µm, 80 g Grace, dry loading, mobile phase, gradient: heptane/EtOAc 100/0 to 50/50) to give 3.28 g of intermediate Q1 in orange oil (67% yield).

^Q1 _R1

Synthesis of intermediate R1.

In a Schlenk flask, to a solution of Q1 (3.28 g; 6.36 mmol) in THF (52 ml) was added an aqueous solution of NH ₃ (30%) (52 ml) at rt. The mixture was stirred at rt. for 26 h and an additional aqueous solution of NH ₃ (10 ml) was added and the mixture was stirred at rt. for 4 h. The mixture was absorbed with EtOAc and saturated aqueous NaCl solution, the layers were separated and the organic layer was dried over MgSO ₄ , filtered and the solvent was removed under reduced pressure to obtain 2.74 g of intermediate R1 as a yellow oil (87% yield ).

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Synthesis of intermediate S1.

NaH (60% in oil) (888 mg; 22.2 mmol) was added in portions to 3-buten-1-ol (30 mL; 354 mmol) at rt. The mixture was stirred at rt. for 30 min before adding dropwise to a solution of R1 (2.74 g; 5.63 mmol) in THF (62 ml) at 0°C. The resulting mixture was stirred at rt. for 1 h and poured into a saturated aqueous solution of NH ₄ Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated and the aqueous layer was extracted with EtOAc (once). The combined organic layers were dried over MgSO ₄ , filtered and the solvent removed under reduced pressure to give a yellow oil. The oil was purified by preparative LC (15-40 µm irregular grain SiOH, 80 g Grace, dry loading, mobile phase, gradient: heptane/EtOAc 100/0 to 20/80) to give 1.06 g of intermediate S1 in as a yellow residue (42% yield).

Synthesis of intermediate T1.

The reaction was performed in two batches of 480 mg of intermediate S1.

The following is the procedure for one batch.

In a Schlenk flask, a solution of S1 (480 mg; 1.08 mmol) and chlordicyclohexylborane (1M in hexane) (216 μl; 216 μmol) in dry dichloroethane (300 ml) was stirred at 80°C and under N ₂ atmosphere for 1 h. Grubbs-Hoveyda 2nd generation catalyst (68 mg; 108 µmol) was added and the mixture was stirred at 120°C for 2 hours.

Two batches were mixed, DMT SiliaBond (2.84 g; 1.73 mmol) was added and the mixture was stirred at rt. within 20 hours

The mixture was filtered through a pad of celite, the celite was washed with EtOAc and the filtrate was evaporated in vacuo to give a brown solid. The brown solid was purified by preparative LC (SiOH 15-40 μm irregular grains, 40 g Grace, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /EtOAc 100/0 to 20/80) to give 610 mg yellow residue (mixture of E and Z isomers, intermediate U1). 310 mg of intermediate U1 was purified by reverse phase chromatography (stationary phase: Nucleodur-Sphinx rp 5 μm 21x150 mm, mobile phase: gradient from 70% formic acid 0.1%, 30% MeCN to 0% formic acid 0.1%, 100% MeCN) to give 195 mg of Intermediate T1 (E-isomer) as a yellow solid (22% yield).

^T1 5

Synthesis of final compound 5.

A mixture of T1 (160 mg; 385 µmol) and iron (129 mg; 2.31 mmol) in acetic acid (21 ml) and water (2.4 ml) was stirred at 80°C for 7 h. Additional iron (129 mg; 2.31 mmol) and the mixture was stirred at 80° C. for 16 h. Additional iron (129 mg; 2.31 mmol) was added and the mixture was stirred at 80° C. for 3 h. The mixture was concentrated in vacuo to give a residue . The residue was taken up in DMF and filtered through a Celite pad. SiliaBond imidazole (12.7 g; 14.7 mmol) was added to the filtrate and the mixture was stirred at rt. for 16 hours. The mixture was filtered through a Celite pad and the filtrate was evaporated in vacuo to give a brown solid. The brown solid was purified by preparative LC (irregular SiOH 15-40 µm, 12 g Grace, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq 97/3/0.3 to 80/20/2) to give 65 mg of an off-white solid. The solid was purified by reverse phase chromatography (stationary phase: X-Bridge-C18 5 μm 30x150 mm, mobile phase gradient: H ₂ O (0.5% NH ₄ CO ₃ )/MeOH 70/30 to 0/100) to give 43 mg of final compound 5 as a white solid (31% yield, E-isomer).

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General scheme for obtaining final products: method 6.

Synthesis of intermediate V1.

Wilkinson's catalyst (58 mg; 62.6 µmol) was added to a solution of U1 (Z/E mixture, 260 mg; 626 µmol) in THF/MeOH (50/50) (66 ml) purged by bubbling N ₂ for 15 min. . The mixture was hydrogenated (7 bar) at rt. over 16 hours. Additional Wilkinson's catalyst (58 mg; 62.6 µmol) was added and the mixture was hydrogenated (7 bar) at rt. for 6 h. The reaction mixture was concentrated in vacuo to give a brown solid. The solid was purified by preparative LC (15-40 µm irregular grain SiOH, 25 g Merck, dry loading, mobile phase, gradient: heptane/EtOAc 100/0 to 50/50) to give 250 mg of intermediate V1 as yellow oil (54% yield).

Synthesis of final compound 6.

A mixture of V1 (238 mg; 359 µmol) and iron (120 mg; 2.16 mmol) in acetic acid (20 ml) and water (2.2 ml) was stirred at 80°C for 6 h. Additional iron (120 mg; 2.16 mmol) and the mixture was stirred at 80° C. for 20 h. Additional iron (120 mg; 2.16 mmol) was added and the mixture was stirred at 80° C. for 5 h. The mixture was concentrated in vacuo to give a residue . The residue was taken up in DMF and filtered through a Celite pad. SiliaBond imidazole (11.1 g; 12.9 mmol) was added to the filtrate and the mixture was stirred at rt. for 16 hours. The mixture was filtered through a Celite pad and the filtrate was evaporated in vacuo to give a brown solid. The solid was purified by preparative LC (irregular SiOH 15-40 µm, 12 g Grace, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq 97/3/0.3 to 80 /20/2) to give 32 mg of an off-white solid. The solid was absorbed with water, triturated and sonicated. The resulting suspension was filtered (glass frit n°5) and washed with Et ₂ O (twice) to give 19 mg of final compound 6 as an off-white solid (15% yield).

General scheme for obtaining final products: method 7.

D1 W1

Synthesis of intermediate compound W1.

NaH (60% in oil) (2.1 g; 52.1 mmol) was added in portions to 3-buten-1-ol (74 ml) at rt. The mixture was stirred at rt. for 30 min before adding dropwise to a solution of D1 (5.97 g; 13.2 mmol) in THF (150 ml) at 0°C. The resulting mixture was then stirred at rt. for 2 h 30 min and poured into a saturated aqueous solution of NH ₄ Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated and the aqueous layer was extracted with EtOAc (once). The combined organic layers were dried over MgSO ₄ , filtered and the solvent removed under reduced pressure to give 6.77 g of a yellow oil. The crude material was purified by preparative LC (irregular SiOH 15-40 µm, 120 g Grace, liquid injection, mobile phase, gradient: heptane/EtOAc 100/0 to

- 11 039955

50/50) to give 5.12 g of intermediate W1 as a yellow oil (83% yield).

Synthesis of intermediate compound X1.

To a solution of W1 (3 g; 6.78 mmol) in CH ₂ Cl ₂ (1.3 L) degassed by bubbling N ₂ for 15 min was added Grubbs' 2nd generation catalyst (578 mg; 678 µmol) at .T. The solution was stirred at rt. over 20 h. DMT SiliaBond (8.89 g; 5.42 mmol) was added and the mixture was stirred at rt. for 20 hours. The reaction mixture was filtered through a Celite pad and the solvent was removed under reduced pressure to give a brown residue which was combined with another batch (0.226 mmol). The combined residue was taken up in MeOH, sonicated and heated to give a precipitate which was filtered off to give 3.2 g of a brown solid. The crude material was purified by preparative LC (irregular SiOH, 15-40 µm, 220 g Grace, liquid injection, mobile phase, gradient: CH ₂ Cl ₂ /EtOAc 100/0 to 50/50) to give 1.7 g fraction 1 as a light brown solid. Fraction 1 was absorbed with MeOH, sonicated and heated to give a precipitate which was filtered off to give 820 mg of fraction 2 as a light brown solid.

The filtrate was concentrated in vacuo to give 590 mg of fraction 3 as a brown residue (crude product X1). Fraction 2 was purified by preparative LC (stationary phase: uncoated spherical silica beads 5 μm 150x30.0 mm, mobile phase, gradient: heptane/EtOAc 85/15 to 0/100) to give 435 mg of intermediate X1 as a yellow solid substances (E-isomer, 15% yield).

Fraction 3 was purified with another batch.

Synthesis of final compound 10.

A mixture of X1 (430 mg; 1.04 mmol) and iron (579 mg; 10.4 mmol) in acetic acid (43 ml) and water (3 ml) was stirred at 50° C. for 4 hours. The mixture was concentrated to dryness. Added DMF. The mixture was sonicated, heated and filtered through a Celite pad and the Celite was washed with hot DMF. SiliaBond imidazole (17.9 g; 20.8 mmol) was added to the filtrate and the mixture was stirred at rt. for 16 hours. The mixture was filtered through celite, the celite was washed with DMF and the filtrate was concentrated in vacuo to give 670 mg of the crude compound. The crude material was purified by preparative LC (irregular SiOH, 15-40 µm, 25 g Merck, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq. 98/2/0.2 to 85/15/ 1.5) to give an off-white solid. The solid was dried at 40° C. under reduced pressure for 20 hours to give 295 mg of final compound 10 as an off-white solid (84% yield).

General scheme for obtaining final products: method 8.

XI _Y1 14

Synthesis of intermediate compound Y1.

Wilkinson's catalyst (103 mg; 111 µmol) was added to a solution of X1 (230 mg; 0.555 mmol) in THF/MeOH (50/50) (40 ml) purged by bubbling N ₂ for 15 min. The mixture was hydrogenated (8 bar) at rt. within 24 hours the Reaction mixture was concentrated in vacuo to obtain a brown residue. The solid was purified by preparative LC (irregular SiOH, 15-40 μm, 12 g Grace, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /EtOAc 100/0 to 90/10) to give 55 mg of intermediate compound Y1 as a yellow residue (24% yield).

Synthesis of final compound 14.

A mixture of Y1 (55 mg; 0.132 mmol) and iron (74 mg; 1.32 mmol) in acetic acid (5.5 ml) and water (0.4 ml) was stirred at 50°C for 20 h. More iron was added (37 mg; 0.66 mmol) and a mixture of

- 12 039955 was stirred at 50° C. for 3 hours. More iron (37 mg; 0.66 mmol) was added and the mixture was stirred at 50° C. for 20 hours. The mixture was filtered through a pad of celite and the celite was washed with acetic acid. More iron (74 mg; 1.32 mmol) was added to the filtrate and the mixture was stirred at 50°C for 88 hours. More iron (74 mg; 1.32 mmol) was added to the filtrate and the mixture was stirred at 80°C for 24 hours Cyclization was not carried out. The mixture was concentrated in vacuo to give a brown solid.

TiCl ₃ (8.60 ml; 10.0 mmol) was added dropwise to a solution of the brown solid in C (19 ml). The mixture was stirred at rt. during the night. The mixture was alkalized by adding K ₂ CO ₃ powder at 0°C. The resulting mixture was filtered through a pad of celite and the celite was washed with a solution of AcOEt/MeOH (8:2). The filtrate was concentrated in vacuo. The crude solid was purified by preparative LC (irregular SiOH, 15-40 µm, 10 g Merck, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq. from 98/2/0.2 up to 85/15/1.5). Fractions containing product were combined and the solvent was removed in vacuo to give 20 mg of final compound 14 (12% yield) as an off-white solid.

General scheme for obtaining final products: method 9.

Synthesis of intermediate compound A2.

Methanesulfonyl chloride (8.4 ml; 108 mmol) was added to a solution of Z1 (14 g; 72.1 mmol), NEt ₃ (20 ml; 144 mmol) and LiCl (4.6 g; 108 mmol) in dry CH ₂ Cl ₂ (980 ml). The mixture was stirred at rt. within 1 hour 30 minutes. Water was added and the layers were separated. The organic layer was washed with water (once), dried over MgSO ₄ , filtered and concentrated in vacuo to give 18.8 g of A2 (96%) as a green oil.

AT 2

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Synthesis of intermediate B2.

Intermediate B2 was prepared using the procedure described for Intermediate C1 (78% yield as a yellow oil).

AT 2

C2

Synthesis of intermediate compound C2.

intermediate

Intermediate C2 was obtained using the procedure described for compound D1 (quantitative yield as a yellow oil).

Synthesis of intermediate compound D2.

Intermediate D2 was prepared using the procedure described for Intermediate W1 (64% yield as a yellow solid).

Synthesis of intermediate compound E2.

A solution of D2 (1 g; 2.12 mmol) in dry CH ₂ Cl ₂ (400 ml) was degassed by bubbling N ₂ for 15 minutes. Grubbs 2nd generation catalyst (181 mg; 212 µmol) was added and the mixture was stirred at rt. over 16 h. DMT SiliaBond (2.78 g; 1.69 mmol) was added and the mixture was stirred at rt. for 16 hours. The mixture was filtered through a Celite pad and the filtrate concentrated in vacuo to give 1.11 g of a brown oil. The crude material was purified by preparative LC (irregular SiOH 15-40 μm, 50 g, Merck, mobile phase, gradient: CH ₂ Cl ₂ /EtOAc 100/0 to 90/10). Fractions containing product were combined and solvent was removed in vacuo to give 386 mg of intermediate E2 (41%, isomer E (96.2%) + isomer Z (3.8%)) as a yellow foam.

Synthesis of final compound 15.

Iron (291 mg, 5.21 mmol) was added to a solution of E2 (386 mg, 0.869 mmol) in acetic acid (36 ml) and water (3 ml). The mixture was stirred at 80° C. for 6 hours. Iron (146 mg; 2.61 mmol) was added and the mixture was stirred at 80° C. for 16 hours. Iron (146 mg; 2.61 mmol) was added again and the mixture was stirred at 80° C. for 5 hours. The mixture was concentrated to dryness. DMF was added, the mixture was filtered through Celite and the Celite was washed with DMF. Siliabond imidazole (18 g; 20.9 mmol) was added to the filtrate and the mixture was stirred at rt. for 72 hours. The mixture was filtered through celite, the celite was washed with DMF and the filtrate was concentrated in vacuo to give 428 mg of a brown solid. The solid was absorbed into CH3CN causing it to precipitate. The precipitate was filtered to give 267 mg of a brown solid. The solid was purified by preparative LC (irregular SiOH 15-40 µm, 10 g Merck, dry loading, mobile phase, gradient: CH ₂ Cl ₂ /MeOH/NH ₃ aq 95/5/0.5 to 85 /15/1.5). Fractions containing product were combined and the solvent was removed in vacuo to give 124 mg of an off-white solid. The solid was purified by reverse phase chromatography (stationary phase: Sunfire-C18 5 µm 19x150 mm, mobile phase, gradient: CH3CN/H2O (formic acid 0.1%) from 5/95 to 50/50) to give 72 mg co- 14 039955 final compound 15 (23% yield) as a white solid.

General scheme for obtaining final products: method 10.

Synthesis of intermediate compound F2.

Intermediate F2 was prepared using the procedures described for Intermediate F1 (E-isomer).

Synthesis of final compound 16.

At k.t. TiCl ₃ (12.3 ml; 14.341 mmol) was added dropwise to a mixture of F2 (300 mg; 0.717 mmol) in THF (30 ml). The mixture was stirred at rt. within 2 hours the Mixture was cooled to 0°C and alkalized with powder K ₂ CO ₃ . The resulting hazy mixture was filtered through a pad of celite and the celite was washed with an AcOEt/CH ₃ OH 8/2 solution. The filtrate was partially evaporated to give 175 mg of the final compound 16 h after filtration of a white solid and drying under reduced pressure at 85° C. (71% yield).

Synthesis of final compound 17.

Received hydrochloride salt with 10 eq. HCl 4 N in dioxane, which was added to a suspension of compound 16 (100 mg; 0.292 mmol) in CH ₃ OH (10 ml). The precipitate was stirred for 3 h, filtered and dried in vacuo at 90° C. overnight. The solid was solubilized in MeOH/CH ₂ Cl ₂ 50/50, CH3CN was added and the solvent was evaporated to give a precipitate of a white solid which was filtered and dried in vacuo at 90° C. to give 47 mg of final compound 17 as the HCl salt (0. 93 HCl, 0.51 H2O, 42% yield).

General scheme for obtaining final products: method 11.

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Synthesis of intermediate H2.

G2 (22.0 g; 72.04 mmol) in THF (100 ml) was added dropwise to a suspension of LiAlH4 (3.28 g; 86.45 mmol) in THF (120 ml) at -20° C. under a stream of N ₂ . The mixture was stirred at 0° C. for 1 hour. 3.5 ml of water was added dropwise, followed by the addition of 3.5 ml of NaOH, 3 H, and 10 ml of water. The resulting mixture was filtered through a Celite pad and the Celite was washed with EtOAc. The filtrate was concentrated under reduced pressure to give (quantitative).

g intermediate H2 as a yellow oil

Synthesis of intermediate 12.

At 0°C, diisopropyl azodicarboxylate (4.0 ml; 20.32 mmol) was added dropwise to a mixture of A1 (5.0 g; 13.55 mmol), H2 (4.28 g; 16.26 mmol) and PPh ₃ (5 .33 g; 20.327 mmol) in THF (100 ml). The mixture was stirred at rt. within 12 hours Added EtOAc and water. The layers were decanted. The organic layer was washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated. The crude material was dissolved in heptane/EtOAc 80/20, the precipitate was filtered off (mainly POPh ₃ ) and the filtrate was purified by chromatography. Purification was performed by flash chromatography through silica gel (15-40 μm, 220 g, heptane/EtOAc 80/20). The pure fractions were collected and evaporated to dryness to give 8.2 g of intermediate 12 (99% yield).

Synthesis of intermediate J2.

(8.2 g; 13.349 mmol) was stirred in NH4OH (100 ml) and THF (100 ml) at RT. within 24 hours the Mixture was evaporated by half under reduced pressure. The residue was absorbed with EtOAc. The organic layer was washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated to give 8.15 g of intermediate J2 (quantitative). The crude material was used directly in the next step.

Synthesis of intermediate compound K2.

NaH (60% in oil) (1.15 g; 28.64 mmol) was added in portions to allyl alcohol (35 ml) at rt under N ₂ flow. The mixture was stirred at rt. for 30 min before adding dropwise to a solution of J2 (4.0 g; 7.26 mmol) in THF (80 ml) at 0°C. The resulting mixture was then stirred at rt. for 2 h 30 min and poured into a saturated solution of NH ₄ Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated and the aqueous layer was extracted with EtOAc (once). The combined organic layers were dried over MgSO ₄ , filtered and the solvent removed under reduced pressure to give 4.7 g of a yellow oil. Purification was performed by flash chromatography on silica gel (15-40 μm, 80 g, CH ₂ Cl ₂ /heptane 65/35). The pure fractions were collected and evaporated to dryness to give 2.65 g of intermediate K2 (69% yield).

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L2

Synthesis of intermediate compound L2.

Before the reaction, the dichloroethane was degassed by bubbling N ₂ through it.

In a Schlenk tube, a solution of K2 (1.3 g; 2.464 mmol) and chlordicyclohexylborane (1M in hexane) (493 µl; 0.493 mmol) in dichloroethane (600 ml) was stirred at 80° C. under N ₂ atmosphere for 1 h. Catalyst was added Grubbs-Hoveyda 2nd generation (609 mg; 0.493 mmol) and the mixture was stirred at 120° C. for 16 h. DMT Siliabond (2.98 g; 1.82 mmol) was added and the mixture was stirred at rt. for 16 hours. The reaction mixture was filtered through Celite and the filtrate was evaporated in vacuo to give 1.6 g, which was combined with another reaction mixture (2.46 mmol) before purification (total weight before purification 3.2 g). Purification was performed by flash chromatography on silica gel (15-40 μm, 80 g, CH ₂ C1 ₂ /CH ₃ OH: 99.5/0.5). The pure fractions were collected and evaporated to dryness to give 0.99 g F1 (E/Z mixture of putative compound, 40% yield) and 0.65 g F2 (starting material K2).

F1 was further purified with achiral SFC (stationary phase: NH ₂ 5 μm 150x30 mm), mobile phase: 92% CO ₂ , 8% MeOH to give 664 mg of intermediate L2 (E-isomer, 27% yield).

Synthesis of intermediate compound M2.

Iron (1.45 g, 26.025 mmol) was added to a mixture of L2 (0.65 g, 1.301 mmol) in acetic acid (15 ml) and water (1.5 ml). The mixture was stirred at 50° C. for 3 h and then filtered through celite with CH ₂ Cl ₂ /MeOH. The filtrate was concentrated under reduced pressure. The compound was purified by flash chromatography through a silica gel column (15-40 µm; 80 g, eluent CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH 96/4/0.5) to give 640 mg. A second purification was carried out by flash chromatography through silica gel (15-40 μm, 40 g, CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 97/3/0.2). The pure fractions were collected and evaporated to dryness to give 240 mg of intermediate M2 (38% yield).

Synthesis of final compound 18.

At 0° C., CF ₃ CO ₂ H (0.455 ml) was added dropwise to a mixture of M2 (100 mg, 0.236 mmol) in CH ₂ Cl ₂ (1 ml). The mixture was stirred at rt. overnight, and then alkalized with a 10% solution of K ₂ CO ₃ in water. The precipitate was filtered off, washed with water and CH3CN and finally dried in vacuo to give 35 mg of final compound 18 (E-isomer, 46% yield).

General scheme for obtaining final products: method 12.

M2 _N2 23

Synthesis of intermediate N2.

A mixture of M2 (140 mg, 0.331 mmol) in THF/CH ₃ OH (50/50) (30 ml) was hydrogenated at 10 bar with Wilkinson's catalyst (61.2 mg, 0.0661 mmol) for 72 h. DMT was added Siliabond (441 mg, 0.264 mmol) and the mixture was stirred at rt. for 18 hours. The mixture was filtered through a pad of celite and the celite was washed with CH ₂ C1 ₂ /CH ₃ OH 95/5. The filtrate was concentrated under reduced pressure. Purification was performed by flash chromatography on silica gel (15-40 μm, 10 g, CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 97/3/0.1). The pure fractions were collected and evaporated to dryness to give 62 mg of intermediate N2 (44% yield) which was used without further purification in the next step.

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Synthesis of final compound 23.

At 0° C., CF ₃ CO ₂ H (0.281 ml, 3.643 mmol) was added dropwise to a mixture of N2 (62 mg, 0.146 mmol) in CH ₂ Cl ₂ (1 ml). The mixture was stirred at rt. during the night. The mixture was alkalized with 10% K2CO3 solution in water. The mixture was extracted twice with CH ₂ Cl ₂ and CH 3 OH (80/20). The organic layer was dried over MgSO ₄ , filtered and the solvent was evaporated. The crude material was absorbed with DMF, 2 g SiO ₂ , 60-200 µm was added and the resulting suspension was evaporated to dryness. This residue was applied to a chromatographic column (to precipitate a solid). Purification was performed by flash chromatography on silica gel (15-40 μm, 25 g, CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 95/5/0.5). Pure fractions were collected and evaporated to dryness to give 20 mg. The fraction was absorbed with CH3CN, the precipitate was filtered off and dried in vacuo to give 18 mg of final compound 23 (38% yield).

General scheme for obtaining final products: method 13.

22 (HC1 salt)

Synthesis of intermediate compound O2.

Intermediate O2 was prepared using the procedures described for Intermediate X1 (E-isomer).

Synthesis of final compound 21.

At k.t. TiCl ₃ (51.5 ml; 60.128 mmol) was added dropwise to a mixture of O2 (1.3 g; 3.006 mmol) in THF (130 ml). The mixture was stirred at rt. for 2 hours. The mixture was cooled to 0°C and then alkalized with K2CO3 powder. The resulting hazy mixture was filtered through a pad of celite and the celite was washed with an AcOEt/CH ₃ OH 8/2 solution. The filtrate was partially evaporated to give 380 mg of final compound 21 (35% yield) after filtration of a white solid and drying in vacuo at 85°C.

Synthesis of final compound 22.

Compound 21 (118 mg, 0.331 mmol) in CH 3 OH (2 ml)+CH ₃ CN (2 ml) was cooled to 10°C. HCl (6 M in isopropanol) (0.16 mL, 0.993 mmol) was added dropwise and the mixture was stirred at rt. within 1 h. The precipitate was filtered off, washed with Et ₂ O and dried in vacuo to give 109 mg of final compound 22 as the HCl salt (0.76 HCl 0.81 H ₂ O, 83% yield).

General scheme for obtaining final products: method 14.

⁰² R2 19

(salt HC1)

Synthesis of intermediate P2.

A mixture of O2 (320 mg; 0.74 mmol), Wilkinson's catalyst (137 mg; 0.148 mmol) in THF/CH3OH (50/50) (45 ml) was hydrogenated at 10 bar at rt. within 20 hours the Solvent was evaporated in vacuum. The crude material was purified by flash chromatography through a silica gel column (15-40 μm; 24 g) in heptane/AcOEt 80/20 to give 310 mg of intermediate P2 (96% yield).

Synthesis of final compound 19.

At k.t. TiCl ₃ (9.5 ml; 11.049 mmol) was added dropwise to a mixture of P2 (0.24 g; 0.552 mmol) in THF (25 ml). The mixture was stirred at rt. for 2 hours. The mixture was cooled to 0°C and then alkalized with K2CO3 powder. The resulting cloudy mixture was filtered through a pad of celite and the celite was washed with an 8/2 AcOEt/CH3OH solution. The filtrate was partially evaporated to give 100 mg of final compound 19 (50% yield) after filtration of a white solid and drying in vacuo at 85°C.

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Synthesis of final compound 20.

Compound 19 (58 mg; 0.162 mmol) in CH ₃ OH (2 ml)+CH ₃ CN (4 ml) was cooled to 5°C. HCl (6 M in isopropanol) (81 μl; 0.486 mmol) was added dropwise and the mixture was stirred at rt. for 1 h. The precipitate was filtered off, washed with diisopropylene ether and dried in vacuo at 90° C. to give 57 mg of final compound 20 as the HCl salt (0.88 HCl 0.04 H2O, 89% yield).

General scheme for obtaining final products: method 15.

Vg aq.

R2

Grubbs-Hoveid catalyst 2nd generation

Synthesis of intermediate compound R2.

At 0°C, diisopropyl azodicarboxylate (3.8 ml; 19.107 mmol) was added dropwise to a mixture of A1 (4.7 g; 12.738 mmol), Q2 (2.27 g; 12.738 mmol) and PPh3 (5 g; 19.107 mmol) in THF (100 ml). The mixture was stirred at rt. within 12 hours Added EtOAc and water. The layers were decanted. The organic layer was washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated.

The crude material was purified by column chromatography through silica gel (15-40 μm; 220 g) in heptane/AcOEt 85/15 to give 5.3 g of intermediate R2 (79% yield).

Synthesis of intermediate compound S2.

Stir R2 (5.3 g; 10.015 mmol) in THF (80 ml) and NH4OH (80 ml) at RT. within 24 hours the Mixture was concentrated under reduced pressure. The residue was absorbed with CH ₂ Cl, the precipitate (inorganic) was filtered off and the filtrate was concentrated under reduced pressure. The crude material was purified by column chromatography through silica gel (15-40 μm; 220 g) in heptane/AcOEt 85/15 to give 3.65 g of intermediate S2 (78% yield).

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Synthesis of intermediate T2.

NaH (1.35 g; 33.88 mmol) was added in portions to allyl alcohol (41 ml) at rt. The mixture was stirred at rt. for 30 min before adding dropwise to a solution of S2 (4 g; 8.597 mmol) in THF (100 ml) at 0°C. The resulting mixture was then stirred at rt. for 2 h 30 min and poured into a saturated aqueous solution of NH4Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated and the aqueous layer was extracted with EtOAc (once). The combined organic layers were dried over MgSO ₄ , filtered and the solvent removed under reduced pressure to give a yellow oil. The crude material was purified by preparative LC (SiOH 15-40 µm irregular grains, 120 g Grace, liquid injection, mobile phase, gradient: heptane/EtOAc 85/15) to give 3.2 g of intermediate T2 as a yellow oil (84 % exit).

Synthesis of intermediate compound U2.

A solution of T2 (1 g; 2.26 mmol) and chlordicyclohexylborane (1 M in hexane) (904 µl; 904.013 µmol) in dry dichloroethane (540 ml) was stirred at 80° C. and under N ₂ atmosphere for 1 h. The mixture was degassed by sparging N ₂ for 15 min, Grubbs-Hoveyda 2nd generation catalyst (141.6 mg; 226 μmol) was added, the mixture was degassed again by sparging N ₂ for 15 min, and then stirred at 120°C for 16 hours 0.25 eq. catalyst and the mixture was stirred at 120° C. for 16 h. DMT Siliabond (5.9 g; 3.616 mmol) was added and the mixture was stirred at rt. for 16 hours. The mixture was filtered through a Celite pad and the filtrate was concentrated in vacuo to give a black oil. The crude compound was purified by preparative LC (irregular SiOH 15-40 μm 80 g, Merck, mobile phase: CH ₂ Cl ₂ /AcOEt 97/3). Fractions containing product were combined and the solvent was removed in vacuo to give 335 mg of intermediate U2 (E-isomer, 36% yield).

Synthesis of final compound 25.

Iron (0.45 g, 8.084 mmol) was added to a mixture of U2 (0.335 g, 0.808 mmol) in acetic acid (24 ml) + water (5 ml). The mixture was vigorously stirred at 50°C for 5 hours.

CH ₂ Cl ₂ was added and the reaction mixture was filtered through a Celite pad and then washed with acetic acid. The solvent was removed under reduced pressure. The crude material was purified by silica gel column chromatography (SiO ₂ 15-40 μm, 25 g) in CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH 96/4/0.5 to give 154 mg of final compound 25 (56% yield) . The compound was crystallized in CH ₃ OH, filtered and dried in vacuo at 90° C. to give 70 mg (25% yield).

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General scheme for obtaining final products: method 16.

J2

Synthesis of intermediate V2.

Intermediate V2 was synthesized using the procedure described for Intermediate K2, starting from 3-butenol (3.9 g, 44% yield).

W2 X2

Synthesis of intermediate compounds W2 and X2.

Added Grubbs' catalyst 2nd generation (236 mg, 0.277 mmol) to a mixture of V2 (1.5 g, 2.77 mmol) in dry CH ₂ Cl ₂ (400 ml). The mixture was stirred at rt. under N ₂ flow for 24 h. DMT Siliabond (3.6 g, 2.216 mmol) was added and the mixture was stirred at rt. for 12 hours. The mixture was filtered through Celite, the Celite was washed with CH ₂ Cl ₂ and the filtrate was evaporated. Purification was performed by flash chromatography on silica gel (15-40 μm, 80 g, CH ₂ C1 ₂ /CH ₃ OH: 99.5/0.5), pure fractions were collected and evaporated to dryness to obtain 0.98 g of a mixture of W2 and X2. The two isomers were separated by achiral SFC (stationary phase: CHIRALPAK IC 5 μm 250x20 mm), mobile phase: 70% CO ₂ , 30% CH3OH) to give 0.805 g of intermediate W2 (E-isomer, 57% yield) and 0.14 g intermediate compound X2 (Z-isomer, 10% yield).

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Synthesis of final compound 26.

Final compound 26 was synthesized according to the procedures described for final compound 18 (1st step: Y2, 0.68 g, 99% yield; 2nd step: 52 mg, 27% yield).

Synthesis of final compound 29.

Final compound 29 was synthesized according to the procedures described for final compound 18 (1st step: Z2, 0.12 g, 100% yield; 2nd step: 8 mg, 9% yield).

General scheme for obtaining final products: method 17.

HCl (6M in 1Ρ1ΌΗ

_CH4OH / _CH4CN

Catalyst Grabos

2nd generation

Synthesis of the intermediate compound VZ.

Allyl bromide (13 ml, 0.15 mmol) was added dropwise to a mixture of A3 (23 g, 0.135 mmol) and K ₂ CO ₃ (28 g, 0.2 mmol) in CH3CN (460 ml). The mixture was stirred at reflux for 4 hours and then concentrated under reduced pressure. The residue was taken up in water and extracted with EtOAc. The organic layers were combined, washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated. The crude material (27 g, 95% yield) was used directly in the next step.

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Synthesis of intermediate C3.

Under N2 atmosphere, DIBAL-H (1.2 M in toluene) (97 ml; 116.5 mmol) was added to a solution of B3 (9.8 g; 46.6 mmol) in dry CH ₂ Cl ₂ (250 ml) at 0 °C. The reaction mixture was stirred at 0°C for 1 h, and then 1 h at room temperature. Water was added, the organic layer was separated from the aqueous layer, dried over MgSO ₄ , filtered and concentrated in vacuo to give 8.4 g of intermediate C3 (99% yield). The crude material was used directly in the next step.

Synthesis of intermediate compound D3.

Intermediate D3 was synthesized using the procedure described for Intermediate R2, starting with C3 (1.9 g, 88% yield).

Synthesis of intermediate compound E3.

Intermediate E3 was synthesized using the procedure described for Intermediate S2, starting from D3 (1.8 g, 93% yield).

Synthesis of intermediate compound F3.

Intermediate F3 was synthesized using the procedure described for Intermediate W1, starting with E3 (0.65 g, 66% yield).

Synthesis of intermediate compound G3.

Intermediate G3 was synthesized using the procedure described for Intermediate X1, starting from Intermediate F3 (E-isomer, 520 mg, 19% yield).

Synthesis of final compound 27.

Final compound 27 was synthesized using the procedure described for final compound 10, starting from intermediate G3 (174 mg, 42% yield).

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Synthesis of final compound 28.

Final compound 28 was synthesized using the procedure described for final compound 20, starting from compound 27 (1.01 HCl, 0.89 H ₂ O, 95 mg, 69% yield).

General scheme for obtaining final products: method 18.

Synthesis of intermediate H3.

A mixture of G3 (600 mg, 1.39 mmol), Wilkinson's catalyst (257 mg; 0.278 mmol) in THF/CH3OH (50/50) (120 mL) was hydrogenated at 12 bar at rt. within 20 hours the Solution was concentrated under reduced pressure. Purification was performed by flash chromatography on silica gel (15-40 μm, 30 g, CH ₂ C1 ₂ /CH ₃ OH: 99.5/0.5). The pure fractions were collected and evaporated to dryness and then crystallized from CH3CN to give 150 mg of intermediate H3 (25% yield).

Synthesis of final compound 32.

A mixture of H3 (150 mg; 0.345 mmol) and iron (190 mg; 3.45 mmol) in acetic acid (13 ml) and water (1.5 ml) was stirred at 50°C for 5 h. CH ₂ Cl ₂ was added and the reaction mixture was filtered through a Celite pad and concentrated in vacuo. The crude material was taken up in DMF, filtered through a celite pad and concentrated. The solid was previously purified by silica gel column chromatography (SiO ₂ 63-200 μm, 80 g) in CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH (98/2/0.1 to 90/10/0, 5). A second purification with achiral SFC (stationary phase: Whelk 01 (c,S) 5 μm 250x21.1 mm, mobile phase: 60% CO ₂ , 40% CH ₃ OH (0.3% iPrNH ₂ )) gave 27 mg of final compound 32 (22% yield).

General scheme for obtaining final products: method 19.

Synthesis of intermediate compound I3.

Intermediate I3 was synthesized using the procedure described for Intermediate T2 (4.2 g, 83%).

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Synthesis of intermediate compound J3.

Intermediate J3 was synthesized using the procedure described for Intermediate F1 (isomer E, 125 mg, 17%).

Synthesis of final compound 30.

Final compound 30 was synthesized using the procedure described for final compound 21 (72 mg, 44% yield).

Synthesis of final compound 31.

Final compound 31 was synthesized using the procedure described for final compound 22 (0.98 HCl, 0.15 H ₂ O, 72 mg, 59% yield).

General scheme for obtaining final products: method 20.

Synthesis of intermediate compound L3.

Allyl bromide (1.7 ml; 19.6 mmol) was added to a solution of K3 (3 g; 17.8 mmol) and K ₂ CO ₃ (2.7 g;

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19.6 mmol) in CH3CN (90 ml). The mixture was stirred at 90°C for 20 h and then filtered. The filtrate was concentrated in vacuo. The crude product was absorbed with CH ₂ Cl ₂ and NaOH aqueous solution, 5%. The layers were separated and the organic layer was dried over MgSO4, filtered and the solvent was removed under reduced pressure to give 3.9 g of intermediate L3 (quantitative). The crude material was used directly in the next step.

Synthesis of intermediate compound M3.

At 0°C, diisopropyl azodicarboxylate (4.8 ml; 24.36 mmol) was added dropwise to a mixture of A1 (6 g; 16.2 mmol), L3 (3.2 g; 15.36 mmol) and PPh ₃ (6.4 g; 24.36 mmol) in THF (120 ml). The mixture was stirred at rt. within 12 hours Added EtOAc and water. The layers were decanted. The organic layer was washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated. Added 20 ml of heptane/AcOEt 70/30 to precipitate most of the formed PPh ₃ O, which is removed by filtration. The crude product was purified by preparative LC (irregular SiOH 15-40 µm, 120 g Interchim, mobile phase: heptane/EtOAc 80/20) to give 8 g of intermediate M3 (88% yield).

Synthesis of intermediate compound N3.

M3 (8.8 g; 15.7 mmol) was stirred in THF (120 ml) and NH4OH (120 ml) at rt. within 24 hours the Mixture was concentrated under reduced pressure. The residue was taken up in CH ₂ Cl ₂ , the precipitate (inorganic) was filtered off and the filtrate was dried over MgSO ₄ , filtered through a Celite pad and concentrated in vacuo. The crude product was purified by preparative LC (irregular SiOH 15-40 µm, 120 g Interchim, mobile phase: heptane/EtOAc 80/20) to give 3 g of intermediate N3 (38% yield).

Synthesis of intermediate compound 03.

NaH (60% in oil) (0.93 g; 23 mmol) was added in portions to allyl alcohol (28 ml) at rt.

The mixture was stirred at rt. for 30 min before adding dropwise to a solution of N3 (2.9 g; 5.85 mmol) in THF (70 ml) at 0°C. The resulting mixture was then stirred at rt. for 2 h 30 min and poured into a saturated aqueous solution of NH ₄ Cl. Added EtOAc and saturated aqueous NaCl solution, the layers were separated and the aqueous layer was extracted with EtOAc (once). The combined organic layers were dried over MgSO ₄ , filtered and the solvent removed under reduced pressure to give a yellow oil. The crude product was purified by preparative LC (irregular SiOH 15-40 µm, 120 g Grace, liquid injection, mobile phase: heptane/EtOAc 80/20) to give 2.4 g of intermediate O3 (87% yield).

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Synthesis of intermediate P3.

The reaction was carried out in three batches.

A solution of 03 (0.8 g; 1.7 mmol) and chlorodicyclohexylborane (1 M in hexane) (0.68 ml; 0.68 mmol) in dry dichloroethane (400 ml) was stirred at 80°C and under N2 for 1 h. The mixture was degassed by N 2 sparging for 15 min, Grubbs-Hoveyda 2nd generation catalyst (110 mg; 0.17 mol) was added, the mixture was degassed again by N ₂ sparging for 15 min, and then stirred at 120 °C for 16 hours. Once again, 0.050 eq. catalyst (49 mg, 0.084 mmol) and the mixture was stirred at 120° C. for 7 hours. DMT Siliabond (3.3 g; 2.03 mmol) was added and the mixture was stirred at rt. for 16 hours. The mixture was filtered through a Celite pad and the filtrate was concentrated in vacuo to give a black oil. The crude product was purified by preparative LC (irregular SiOH 15-40 µm, 80 g Interchim, mobile phase: heptane/EtOAc 65/35) to give 190 mg of intermediate P3 (isomer E, 25% yield).

Synthesis of final compound 36.

At k.t. TiCl ₃ (19.3 ml; 22.5 mmol) was added dropwise to a mixture of P3 (500 mg; 1.125 mmol) in THF (90 ml). The mixture was stirred at rt. for 2 hours At 0°C the mixture was alkalized with powder K ₂ CO ₃ . The resulting hazy mixture was filtered through a pad of celite and the celite was washed with CH ₂ Cl ₂ /CH ₃ OH (90/10) solution. The filtrate was concentrated under reduced pressure. The residue was taken up in MeOH. The white solid was filtered off and dried in vacuo. The product was purified by preparative LC (irregular SiOH 15-40 µm, 40 g Interchim, mobile phase CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH 98/2/0.1) to give 140 mg of final compound 36 (34 % exit).

LCMS methods.

General procedure VDR2 (for methods V300xV30xx.olp).

LC measurements were performed using an Acquity (Waters) UPLC system containing a binary pump with a degasser, an autosampler, a diode array detector (DAD) and a column as indicated in the respective methods below, the column being maintained at 40°C. The flow from the column was directed to the MS detector. The MS detector was equipped with an electrospray ionization source. The capillary needle voltage was 3 kV and the source temperature in the Quattro (a triple quadrupole mass spectrometer from Waters) was maintained at 130°C. Nitrogen was used as the nebulizer gas. Data collection and processing was performed using the MassLynx-Openlynx data acquisition and processing system from Waters-Micromass.

Method V3018V3001.

In addition to the general VDR2 procedure: reverse phase UPLC was run on a Waters Acquity BEN (bridged ethylsiloxane/silica gel) C18 column (1.7 µm, 2.1 x 100 mm) with a flow rate of 0.343 ml/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were used to run under gradient conditions from 84.2% A and 15.8% B (hold for 0 .49 min) to 10.5% A and 89.5% B in 2.18 min with a hold of 1.94 min and return to initial conditions in 0.73 min with a hold of 0.73 min. An injection volume of 2 μl was used. The cone voltage was 20 V for the positive and negative ionization modes. Mass spectra were obtained by scanning from 100 to 1000 in 0.2 s using a delay time between scans of 0.1 s.

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Table 1. Compounds of formula (I)

No. 1 2 3 _{THE STRUCTURE} ^_ _{_ _} χ^ Mass, exact value 324.1 326.1 322.1 The resulting mass value [M+H] 325 327 323 Retention time for LCMS, method 1.96V3018V3001 2.02, V3018V3001 2.3V3018V3001 Synthesis method Method 1 Method 2 Method 3 NMR X NMR (500 MHz, DMSO-dg) δ 10.06 (s, 1H), 7.36 (s, 1H), 7.20 (t, J=7.9 Hz, 1H) , 6.766.93 ( m, 2H), 6.05 (s, 1H), 5.94 (dt, J=6, 0, 16.1 Hz, 1H), 5.43-5.64 (m, ZN), 4.84 (s, 2H) , 4.40-4.66 (m, 4H) ² H NMR (400 MHz, dmso-d _b ) δ 10.04 (br. s, 1H) , 7.49 (s, 1H ) , 7.17 (t, J=7.9 Hz, 1H) , 6, 69-6.83 (m, 2H) , 6.15 (s, 1H) , 5.54 (s, 2H) , 4 .89 (s, 2H) , 4.28 (t, J=6.3 Hz, 2H) , 4.14 (t, J=6.6 Hz, 2H) , 1.341.62 (m, 4H) (500 MHz, DMSO-s1 ₆ ) δ 10.11 (br.s, 1H) , 7.22 (t, J=7.6 Hz, 1H) , 7.12 (br.s, 1H) , 7, 09 (d, J=7.6 Hz, 1H) , 7.05 (d, 0=7.6 Hz, 1H) , 5.74 (s, 1H) , 5.51-5.63 (m, ZN ) , 5.03 (td, J=7.2, 14.9 Hz, 1H) , 4.93 (s, 2H) , 4.15-4.26 (m, 2H) , 3.15-3, 23 (m, 2H), 2.242.33 (m, 2H) 4 νη ₂ νΑν ^Η0 X 1 1 Λ Г'|ХХс \ 324.2 325 2.36V3018V3001 Method 4 41 NMR (500 MHz, DMSO-ds) δ 10.20 (br. s, 1H) , 7.34 (s, 1H) , 7.21 (t, J=7.3 Hz, 1H) . 7.06 (d, A7.3 Hz, 1H), 6.97 (d, J=7.3 Hz, 1H), 5.59 (s, 2H), 4.93 (s, 2H), 3.91-4.03 (m, 2H), 2.55-2.62 (m, 2H), 1.59-1.71 (m, 2H), 1.12-1.27 (m, 4H) 5 νη ₂ ^ΗΟ Χ L 1 Ν X/ 339.1 340 1.85V3018V3001 Method 5 41 NMR (400 MHz, DMSO-b ₆ ) δ 9.91 (br. s, 1H) , 8.05 (d, 0=4.6 Hz, 1H) , 7.38 (d, 0^=8, 6 Hz, 1H) , 7.27 (dd, 0=4.6, 8.6 Hz, 1H) , 6.20 (s, 1H) , 5.82 (dt, 0=7.0, 15.6 Hz, 1H) , 5.68 (dt, J=7,l, 15.6 Hz, 1H) , 5.46 (s, 2H) , 4.91 (s, 2H) , 4.33 (d, 0 \u003d 7.1 Hz, 2H), 4.12-4.27 (m, 2H), 2.36-2.46 (m, 2H) 6 νη ₂ ,ΝχΧ non* 1 η Ν'Χ/'Ό οΧ 341.1 342 1.85V3018V3001 Method 6 41 NMR (400 MHz, DMSO-ds) δ 9.92 (s, 1H), 8.09 (d, J=4.0 Hz, 1H), 7.43 (d, J=8,l Hz, 1Н), 7.30 (dd, J=4.0, 8.1 Hz, 1H), 6.40 (s, 1H), 5.51 (s, 2H), 4.96 (s, 2H), 4.15 (t, 0=6.1 Hz, 2H), 3.97-4.10 (m, 2H), 1.81-1.91 (m, 2H), 1.681.78 (m, 2H), 1.52-1.65 (m, 2H)

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7 8 10 eleven 12 13 nh ₂ 'G ^N m La lI° nh ₂ n^An iox LL ? nh ₂ N^n ho^ I if N^-^nX i ^н x X ' nh ₂ A^^Ln ^H0 XL ll V nh ₂ N^^m ^H °x/Ot \ Chu ' nh ₂ X nh ₂ AX^ ^{n H0} X JU l ¢¢0 φ o ₄ 354.1 356.1 323.2 338.1 352.2 354.1 356.1 355 357 324 339 353 355 357 1.84V3018V3001 1.86V3018V3001 2.36V3018V3001 2.22V3018V3001 2.33V3018V3001 2.04, V3018V3001 2.1, V3018V3001 Method 1 Method 2 Method 1 Method 2 Method 7 Method 1 Method 1 Method 2 X NMR (400 MHz, DMSO-s1 ₆ ) δ 10.03 (br. s, 1H) , 7.36 (d, J=2.0 Hz, 1H) , 6.91 (d, J=8, l Hz, 1H) , 6.80 (dd, J=2.0, 8.1 Hz, 1H) , 6.06 (dt, J=6.0, 16.2 Hz, 1H) , 5.93 ( s, 1H) , 5.54 (s, 2H) , 5.41 (dt, J=5.6, 16.2 Hz, 1H) , 4.78 (s, 2H) , 4.504.67 (m, 2H ) , 4.28-4.48 (m, 2H) , 3.71 (s, 3H) X NMR (400 MHz, DMSO-dg) δ 10.03 (br. s, 1H) , 7.54 ( br.s, 1H) , 6.87 (d, J=8.l Hz, 1H) , 6.76 (d, J=8.l Hz, 1H) , 6.24 (br.s, 1H) , 5.53 (br.s, 2H) , 4.82 (br.s, 2H) , 4.19-4.32 (m, 2H) , 4.01-4.16 (m, 2H) , 3. 68 (br. s, 3H), 1.34-1.60 (m, 4H) - H NMR (400 MHz, DMSO-dg) δ 9.73 (s, 1H), 7.57 (s, 1H) , 7.16 (t, J=7.6 Hz, 1H) , 7.02 (d, J=7.6 Hz, 1H) , 6.89 (d, J=7.6 Hz, 1H) , 5 .33 (t, J=6.6 Hz, 1H), 5.25 (s, 1H), 5.15 (s, 2H) , 4.85 (s, 2H) , 2.91 (sq., J \u003d 6.6 Hz, 2H) , 2.58-2.72 (m, 2H) , 1.59-1.85 (m, 2H) , 0.96-1.21 (m, 4H) - H NMR (400 MHz, DMSO-dg) δ 10.05 (br. s, 1H), 7.20 (t, J=7.8 Hz, 1H), 6.92 (d, J=7.8 Hz, 1H ) , 6.89 (s, 1H) , 6.77 (d, J=7.8 Hz, 1H) , 6.12 (s, 1H) , 5.50-5.64 (m, 3H) .5 .35 (dt, J=4.5, 16.2 Hz, 1H), 4.87 (s, 2H), 4.58 (d, J=4.5 Hz, 2H), 4.24 (t, J=5.1 Hz, 2H), 2.21-2.32 (m, 2H) ² H NMR (500 MHz, DMSO-dg) δ 10.07 (s, 1H) , 7.32 (s, 1H ) , 7.18 (t, J=7.7 Hz, 1H) , 6.82 (d, J=7.7 Hz, 1H), 6.79 (dd, J=l.7, 7.7 Hz , 1H) , 5.98 (dt, J=7.7, 15.5 Hz, 1H) , 5.80 (s, 1H) , 5.56 (s, 2H) , 5.51 (td, J= 5.8, 15.5 Hz, 1H) , 4.84 (s, 2H) , 4.71 (d, C=5.8 Hz, 2H) , 3.96 (t, J=7.7 Hz, 2H), 1.922.07 (m, 2H), 1.49-1.64 (m, 2H) - H NMR (400 MHz, DMSO-dg) δ 10.05 (br. s, 1H), 6.94 (s, 1H) , 6.47 (s, 1H) , 6.39 (s, 1H) , 6.05 (s, 1H) , 5.87 (dt, J=6, 0, 15.7 Hz, 1H), 5.45-5.68 (m, 3H) , 4.79 (s, 2H) , 4.41-4.61 (m, 4H) , 3.64 (s, 3H) X NMR (500 MHz, DMSO-dg) δ 10.05 (s, 1H) , 7.08 (s, 1H) , 6.24-6.44 (m, 2H) , 6.16 (s, 1H), 5.55 (s, 2H), 4.84 (s, 2H) , 4.25 (t, U=6.3 Hz, 2H) , 4.13 (t, J=6.8 Hz, 2H) , 3.64 (s, ZN) , 1.47-1.62 (m, 2H) , 1.29-1.46 (m, 2H) 14 nh ₂ ^Η °χ ^Ν χ5 _ kgAX 340.2 341 2.23V3018V3001 Method 8 ² H NMR (400 MHz, DMSO-Mb) δ 10.06 (s, 1H), 7.10-7.30 (m, 2H), 6.89 (d, J=7.6 Hz, 1H) , 6.81 (dd, J=2.0, 7.6 Hz, 1H) , 6.25 (s, 1H), 5.58 (s, 2H), 4.86 (s, 2H) , 3, 99-4.12 (m, 4H), 1.33-1.51 (m, 6H)

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16 17 18 19 20 21 nh ₂ ^h °%xS V ³ nh ₂ a T^N ^H °—Jl if -a: ¹ nh ₂ l ^h °—G if - xr ³ nh ₂ N ^h °—L nh ₂ ^but % D J. nh ₂ ^ΗΟ %' ^Ν Χχ _ .0 ³ nh ₂ ho^ X Λ V ³ 368.1 342.1 342.1 323, 1 358.1 358.1 356.1 369 343 343 324 359 359 357 2.25V3018V3001 2.04, V3018V3001 2.04, V3018V3001 1.78V3018V3001 2.27V3018V3001 2.27V3018V3001 2.21, V3018V3001 Method 9 Method 10 Method 10 Method 11 Method 14 Method 14 Method 13 ² H NMR (400 MHz, dmso-ds) δ 10.07 (s, 1H) , 6.52 (s, 1H) , 6.45 (s, 1H) , 6.33 (s, 1H) , 6 .09 (s, 1H) , 5.46-5.68 (m, ZN) , 5.34 (dt, 0=5.2, 15.9 Hz, 1H) , 4.83 (s, 2H) , 4.54 (d, 0=5.2 Hz, 2H) , 4.24 (t, 0=5.3 Hz, 2H) , 3.68 (s, ZN) , 2.21-2.31 (m , 2H) ² H NMR (500 MHz, DMSO-ds) δ 10.13 (br.s, 1H), 7.58 (dd, u=1.Ί, 7.9 Hz, 1H) , 7.14 ( dd, 0=8.3, 10.9 Hz, 1H), 6.73-6.94 (m, 1H), 5.88-6.13 (m, 2H), 5.41-5.75 ( m, 3H), 4.83 (s, 2H) , 4.61 (d, 0=5.4 Hz, 2H) , 4.56 (d, 0=6.3 Hz, 2H) 10 NMR (500 MHz , dmso-a ₆ ) δ 11.15 (br. s, 1H), 7.70 (dd, 0=1.9, 7.9 Hz, 1H), 6.96-7.53 (m, 2H) , 6.80-6.93 (m, 1H), 6.37 (s, 1H) , 6.24 (dt, 0=6.3, 15.8 Hz, 1H) , 5.63 (dt, 0 =5.9, 15.8 Hz, 1H), 4.77-4.99 (m, 4H), 4.58 (d, 0=6.3 Hz, 2H) ² H NMR (500 MHz, dmso- ds) δ 10.01 (s, 1H) , 6.93 (t, 0=7.7 Hz, 1H) , 6.74 (br.s, 1H) , 6.40-6.58 (m, 2H ) , 6.30 (s, 1H) , 6.02 (br.s, 1H) , 5.83 (dt, 0=5.4, 16.0 Hz, 1H), 5.58 (br.s, 2H), 5.36-5.50 (m, 1H), 4.74 (s, 2H), 4.49 (d, 0=5.4 Hz, 2H), 3.56-3.87 (m , 2H) 10 NMR (5 00 MHz, DMSO-ds) δ 9.49 (br. s, 1H), 7.44 (d, 0=7.9 Hz, 1H), 7.14 (dd, 0=8.2, 11.4 Hz, 1H) , 6.84-6.97 (m, 1H) , 6.34 (s, 1H) , 5.69 (s, 2H) , 4.84 (s, 2H) , 4.034.17 (m , 4H), 1.31-1.57 (m, 6H) S NMR (500 MHz, DMSO-ds) δ 11.04 (br. s, 1H), 7.46 (dd, 0=1.9, 8.2 Hz, 1H) , 6.86-7.32 (m, ZN), 6.78 (s, 1H), 4.94 (s, 2H) , 4.41 (t, 0=6.6 Hz, 2H), 4.09 (t, 0=6.9 Hz, 2H), 1.63-1.73 (m, 2H), 1.53-1.62 (m, 2H), 1.36 -1.49 (m, 2H) 41 NMR (500 MHz, DMSO-ds) δ 10.09 (br. s, 1H), 6.99-7.23 (m, 2H), 6.92 (br. s, 1H) , 6.15 (s, 1H), 5.53-5.73 (m, 3H), 5.23-5.48 (m, 1H) , 4.85 (s, 2H) , 4 .67 (d, 0=4.4 Hz, 2H) , 4.24 (t, 0=4.6 Hz, 2H) , 2.212.35 (m, 2H) 22 nh ₂ ^HCI 356.1 357 2.21, V3018V3001 Method 13 41 NMR (500 MHz, DMSO-ds) δ 11.11 (br. s, 1H), 7.10-7.42 (m, ZN) , 7.07 (dd, 0=1.9, 8.2 Hz, 1H) , 6.85-6.99 (m, 1H) . 6.64 (s, 1H) , 5.80 (dt, 0=7.2, 15.6 Hz, 1H) , 5.59 (dt, 0=5.7, 15.6 Hz, 1H) , 4 .98 (s, 2H) , 4.68 (d, 0=5.7 Hz, 2H) , 4.414.59 (m, 2H), 2.38-2.50 (m, 2H)

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23 nh ₂ ^Xn ^but % 325.2 326 1.83V3018V3001 Method 12 H NMR (500 MHz, DMSO-a ₆ ) δ 9.97 (br. s, 1H) , 7.05 (s, 1H) , 6.90 (t, J=7.7 Hz, 1H) , 6, 42 (d, J=7.7 Hz, 1H), 6.38 (d, J=7.7 Hz, 1H) , 6.35 (s, 1H) , 5.33-5.58 (m, W ), 4.77 (s, 2H), 4.17 (t, o=6.8 Hz, 2H), 3.20 (sq., J=6.4 Hz, 2H), 1.42-1, 52 (m, 2H), 1.32-1.41 (m, 2H) 24 O YH· -P OO I 344.1 345 2.08.V3018V3001 Method 14 ^: H NMR (500 MHz, dmso-a ₆ ) δ 10.04 (br. s, 1H), 7.77 (d, o=7.6 Hz, 1H), 7.10 (dd, o=8, 7, 11.2 Hz, 1H) , 6.78 (br.s, 1H) , 6.28 (s, 1H) , 5.56 (s, 2H) , 4.86 (s, 2H) , 4.284, 47 (m, 2H), 4.04-4.23 (m, 2H) , 1.52-1.66 (m, 2H) , 1.311.50 (m, 2H) 25 nh ₂ N^Xm but^ XJ X 338.1 339 2.13V3018V3001 Method 15 ² H NMR (500 MHz, DMSO-s1 _b ) δ 10.07 (br. s, 1H), 7.18 (t, J-7.9 Hz, 1H) , 7.14 (s, 1H) , 6 .73-6.87 (m, 2H), 5.96 (dt, o=5.0, 15.7 Hz, 1H) , 5.90 (s, 1H) , 5.57 (s, 2H) . 5.37 (dt, o=5.8, 15.7 Hz, 1H) , 4.86 (s, 2H) , 4.58 (d, o=5.8 Hz, 2H) , 4.22 (t , J=5, 0 Hz, 2H) , 2.272.42 (m, 2H) 26 nh ₂ ^Η °Α ^Ζ X l X 337.2 338 2.02, V3018V3001 Method 16 ² H NMR (500 MHz, dmso-a ₆ ) δ 10.04 (br. s, 1H), 6.95 (t, o=7.2 Hz, 1H), 6.51 (d, J=7, 2 Hz, 1H) , 6.46 (dd, J=l.3, 7.2 Hz, 1H) , 6.40 (s, 1H) , 6.16 (s, 1H), 5.97 (t, 0=6.3 Hz, 1H), 5.59 (s, 2H), 5.43 (dt, J=6.3, 15.5 Hz, 1H), 5.22 (dt, o=5.0 , 15.5 Hz, 1H) , 4.76 (s, 2H) , 4.23 (t, o=5.2 Hz, 2H) , 3.56 (t, J=5, 0 Hz, 2H) , 2.182.30 (m, 2H) 27 νη ₂ Ν-Χ > _ν ηοΑ ^ζ X Λ V ⁵ F 356.1 357 2.30V3018V3001 Method 17 ^: H NMR (500 MHz, DMSO-ds) δ 10.11 (br. s, 1H), 6.77 (d, o=9.1 Hz, 1H), 6.71 (s, 1H) , 6, 63 (dt, 0=2.2, 9.1 Hz, 1H) , 6.15 (s, 1H) , 5.64 (s, 2H) , 5.57 (dt, J=6.6, 15, 8 Hz, 1H), 5.34 (dt, J=5.3, 15.8 Hz, 1H), 4.89 (s, 2H), 4.59 (d, J=4.7 Hz, 2H) , 4.25 (t, 0=5.3 Hz, 2H), 2.20-2.34 (m, 2H) 28 νη ₂ Ν~Χ^ _ν ^η °4 ^ζ X ι ' ^hci F 356.1 357 2.31, V3018V3001 Method 17 ² H NMR (500 MHz, DMSO-dg) δ 11.09 (br. s, 1H) , 7.15 (br. s, 2H) , 6.82 (d, o=8.8 Hz, 1H) . 6.56-6.70 (m, ZN), 5.71 (dt, o=6.3, 15.6 Hz, 1H), 5.52 (dt, J=5.4, 15.6 Hz, 1H), 4.99 (s, 2H) , 4.62 (d, u=5.4 Hz, 2H) , 4.50 (t, J=4 _r l Hz, 2H) , 2.422.47 (m, 2H)

- 31 039955

29 thirty 31 32 33 nh ₂ N^A _m ^H0 D D JL AY^ nh ₂ ^but D JL AV ^A F nh ₂ Ν'-L but </ AA Ay ⁴ ^ F nh ₂ yiA 4 F nh ₂ N ~ηΛ _Ν but </ AA Ν '^^'ο-η v ^A F 337.2 342.1 342.1 358.1 344.1 338 343 343 359 345 2.05, V3018V3001 2.08, V3018V3001 2.08, V3018V3001 2.32V3018V3001 2.14V3018V3001 Method 16 Method 19 Method 19 Method 18 Method 14 41 NMR (500 MHz, DMSO-s1 ₆ ) δ 10.08 (br. s, 1H), 6.96 (t, D=7.7 Hz, 1H), 6.45-6.57 (m, 2H ), 6.42 (s, 1H), 6.20 (s, 1H), 5.92 (t, J=6.0 Hz, 1H), 5.65 (s, 2H), 5.31-5 .48 (m, 1H), 5.08-5.26 (m, 1H), 4.75 (s, 2H), 4.31 (t, J=5.8 Hz, 2H), 3.41- 3.48 (m, 2H) , 2.12-2.28 (m, 2H) ^X H NMR (500 MHz, DMSO-ds) δ 10.10 (br. s, 1H) , 7.23 (s, 1H) , 6.78 (d, J=8.8 Hz, 1H) , 6.62 (d, J=8.8 Hz, 1H) , 6.08 (s, 1H) , 5.8 4 (dt , J=5.0, 15.8 Hz, 1H), 5.71 (dt, J=5.0, 15.8 Hz, 1H), 5.60 (s, 2H), 4.86 (s, 2H), 4.63 (d, D=5.0 Hz, 2H), 4.54 (d, u=5.0 Hz, 2H) 41 NMR (500 MHz, DMSO-dJ δ 11.12 (br. s, 1H) , 7.34 (s, 1H) , 7.15 (br.s, 2H), 6.85 (d, J=10.4 Hz, 1H) , 6.64 (d, J=10 .4 Hz, 1H) , 6.44 (s, 1H) , 6.07 (dt, J=5.9, 15.8 Hz, 1H) , 5.70 (dt, J=5.9, 15, 8 Hz, 1H), 4.95 (s, 2H) , 4.84 (d, J=5.9 Hz, 2H) , 4.64 (d, J=5.9 Hz, 2H) 41 NMR (500 MHz, DMSO-Mb) δ 10.13 (br. s, 1H), 7.05 (s, 1H), 6.71 (d, J=9.1 Hz, 1H), 6.67 (d, 4 = 9.1 Hz, 1H), 6.28 (s, 1H), 5.63 (s, 2H), 4.87 (s, 2H), 3.85-4.18 (m, 4H), 1 .21-1.59 (m , 6H) ^g H NMR (500 MHz, flMCO-d ₆ ) δ 10.09 (br. s, 1H), 7.33 (s, 1H), 6.66 (d, 4=9.1 Hz, 1H ) , 6.59 (d, 4=9.1 Hz, 1H), 6.17 (s, 1H), 5.58 (s, 2H), 4.90 (s, 2H), 4.29 (t , 4=6.5 Hz, 2H) , 4.15 (t, 4=6.5 Hz, 2H) , 1.48-1.59 (m, 2H), 1.36-1.46 (m, 2H) 34 nh ₂ ho% ^/NxS 339.2 340 2.06, V3018V3001 Method 12 ^g H NMR (500 MHz, flMCO-d ₆ ) δ 10.05 (s, 1H), 6.83-7.00 (m, 1H), 6.62 (s, 1H), 6.50 (d, 4=8.2 Hz, 1H), 6.43 (dd, 4=1.6, 8.2 Hz, 1H) , 6.36 (s, 1H) , 5.72 (t, J=6.6 Hz, 1H), 5.60 (s, 2H), 4.76 (s, 2H), 4.07 (t, J=5.7 Hz, 2H) , 2.95 (sq., J=6, 6 Hz, 2H), 1.12-1.50 (m, 8H)

- 32 039955

Biological activity of the compounds of formula (I).

Description of biological activity assays.

Assessment of TLR7 and TLR8 activity.

The ability of the compounds to activate human TLR7 and TLR8 was evaluated in a reporter gene assay using HEK293 cells transiently transfected with a TLR7 or TLR8 expression vector and an NFkB-Iuc reporter construct. Briefly, HEK293 cells were grown in culture medium (DMEM enriched with 10% FCS and 2 mM glutamine). For transfection of cells in 15 cm dishes, cells were detached with trypsin-EDTA, transfected with a mixture of CMV-TLR7 or TLR8 plasmids (1,700 ng), NFkB-Iuc plasmids (850 ng), and transfection reagent, and incubated for 48 h at 37°C in a humidified atmosphere 5 % CO ₂ . The transfected cells were then washed in PBS, separated with trypsin-EDTA and resuspended in medium at a density of 1.25x105 cells/ml. Forty microliters of cells were then dispensed into each well in 384-well plates already containing 200 nl of the compound in 100% DMSO. After 6 hours incubation at 37° C., 5% CO ₂ , luciferase activity was determined by adding 15 μl of Steady Lite Plus substrate (Perkin Elmer) to each well and readings were taken on a ViewLux ultraHTS microplate reader (Perkin Elmer). Dose-response curves were constructed based on quadruple measurements. For each compound, lowest effective concentration (LEC) values were determined, defined as the concentration that produces an effect that is at least twice the assay tolerance.

In parallel, a similar compound dilution series (200 nl of compound in 100% DMSO) was used with 40 μl per well of cells transfected with the NFkB-Iuc reporter construct alone (1.25x105 cells/ml). Six hours after incubation at 37° C., 5% CO ₂ , luciferase activity was determined by adding 15 μl of Steady Lite Plus substrate (Perkin Elmer) to each well and readings were taken on a ViewLux ultraHTS microplate reader (Perkin Elmer).

Back-screen data were recorded as LEC.

Measurement of Interferon Production in Human PBMCs Activation of human TLR7 results in increased production of interferon by plasmacytoid dendritic cells present in human blood. The ability of a compound to induce interferon production was assessed by determining interferon in conditioned media from peripheral blood mononuclear cells (PBMCs). The presence of interferon in the samples was determined using an interferon detection reporter cell line stably expressing the reporter construct with interferon-stimulated response elements (ISRE)-luc. The ISRE element with the sequence TAGTTTCACTTTCCC is highly sensitive to the transcription factor STAT1-STAT2-IRF9, which is activated when IFN-I binds to the IFN receptor. Briefly, PBMCs were prepared from buffy coats from at least two donors using a standard Ficoll centrifugation protocol. The isolated PBMCs were resuspended in RPMI medium supplemented with 10% human AB serum and 2x10 ⁵ cells/well were dispensed into 384-well plates containing compounds (total volume 70 μl). After overnight incubation, 10 μl of the supernatant was transferred to 384-well plates containing 5x10 ³ HEK-ISRE-luc cells/well in 30 μl (inoculated the day before). After 24 hours of incubation, activation of ISRE elements was determined by performing a luciferase activity assay using 40 μl/well Steady Lite Plus substrate (Perkin Elmer) and detected using a ViewLux ultraHTS microplate reader (Perkin Elmer). The stimulatory activity for each compound on HEK-ISRE-luc cells was presented as LEC. LEC, in turn,

- 33 039955 indicates the degree of activation of ISRE when transferring a certain amount of culture medium PBMC.

Recombinant interferon alfa-2a (Roferon-A) was used as a standard control compound.

The LEC values for the compounds in Table. 2 on HEK293 TLR8-NFB-luc and HEK293 NFkB-Iuc were greater than the highest concentration tested (>10 μM for compound 4 and >25 μM for all other compounds).

Table 2. Compounds of formula (I) n is the number of experiments performed

No. STRUCTURE HEK293 TLR7-NFBluc (LEC; µm) n PBMC HEK-ISREluc (LEC; µm) n 1 L/o 2.88 8 0.30 8 2 nh ₂ N-Χ Chu 4.47 1 0.93 2 3 about t 0.27 2 0.033 4 4 nh ₂ n^A _n aul Chg^ 2.38 1 2.56 2 5 νη ₂ N^A _n ⁿ °у X у oL/ 2.14 1 0.082 2 6 nh ₂ ^H0 Y 1 LL OU 1 1 0.16 2 7 <4 o o o— yy O 3, 88 1 0.29 2 8 nh ₂ nXn ° XX 1 7.1 1 0.58 2 9 nh ₂ N-An ^H0 1A _n a _x L ^H 4X^ 11, 09 1 10, 87 2

- 34 039955

10 nh ₂ ^H0 ^ 1 i /x V 0.78 2 0.25 4 eleven nh ₂ ξ V 1.25 1 0.45 2 12 nh ₂ but/ L ϊ V/ °\ 1, 08 1 0.3 2 13 nh ₂ ^HO ^ JA V ^J 1.71 2 0.15 2 14 nh ₂ N-vAm but/ XA 4^4/°^>\ and 7.01 2 2.4 2 15 T o o w 0.18 1 0.04 2 16 nh ₂ n-^n but-r x 2.02 2 0.47 2 17 nh ₂ Nr^N io—( L £ N'^^'0-I ¹¹⁰¹ 4 1.88 2 0.39 4 18 nh ₂ ^ho ^ Uo-, L η I V ⁵ 8.02 1 1, 11 2 19 nh ₂ ^HO A ^Z D A I? ³ 14.96 2 2.22 4

- 35 039955

20 nh ₂ ^Η °^ώ - 4 ^s 4.99 1 1, 61 2 21 nh ₂ ⁿ °h JA %? 0.9 1 0.25 3 22 nh ₂ XJ 1^°4 , HCI 1, 83 1 0.39 2 23 O 16, 74 2 8.98 4 24 nh ₂ ^h °—L ϊ AX 2.14 4 25 nh ₂ N^^-n HO^ X 1 I? ³ 1.92 1 0.53 2 26 nh ₂ HO—XJ X"5 1.88 2 0.37 4 27 5 Y-Ι Oh oh 1.35 1 0.14 2 28 ^Ν -Χ^ Μ ^ho ^ X a N-%^0^ .on ιΓ^Γ F 0.91 1 0.15 2 29 nh ₂ N-^Λμ ^H °X ^Z J i N'^^'O^zA χΰ 1.14 2 0.48 2

- 36 039955

thirty nh ₂ ho—LX ν° ^Λ F 0.64 1 0.15 2 31 νη ₂ ^Η °—L if F 1.19 1 0.15 2 32 νη ₂ ^ΗΟ Α ^Ζ X JL n4^ ₀ Q> F 2.92 2 0.49 2 33 νη ₂ Ν^Λ ^Η °—Jl if ν ^Λ F 2.77 1 0.45 2 34 νη ₂ HO—JJ V ^J 8.1 2 2.11 2 35 ν-^ν but—Jl if At 17.43 1 1.98 2 36 nh ₂ Λ io—1 if ? l 13, 75 1 1, 63 4 U5

Claims (5)

CLAIM 1. A compound selected from the group consisting of: nh ₂ N^'n ^HO X L 1 N^^O > V nh ₂ n^X _n ^HO X 1 1 - Lx X oyy o nh ₂ n^X _n ^ηο ΛΧα _ο ^ LI 1 nh ₂ N^X _n ^H0 XL 1 I ^H Jx V νη ₂ ^η °Χ;Δ χχ> nh ₂ hoX ^/N ^X nh ₂ // ^N Xn ^HO XJA V ^J °\ νη ₂ ^Η °ΧΧχ YyX nh ₂ ^h °—Г if - w nh ₂ nX'n ^ho XX XyS νη ₂ Ν-^Χμ ⁿ ° X' X j •ϊ nh _{2 but} xD X ^s nh ₂ bx νη ₂ ^Η °ΧΧχ . . Dz nh ₂ Xn ho—JL if N^^Ol 1/ nh ₂ Y'-T^M ⁿ °—E if a: ¹ νη ₂ N-^Χν ^η °χ ^ζ X j nh ₂ no^D Dz nh ₂ N-^Xn ^h °a ^z 11 V ⁵ F νη ₂ ^Η °χ ^/Ν χ5ι , ^{ns |} ιΓχ ^ο F nh ₂ ^Η °χ ^/Ν χ5ί De nh ₂ n-Xn h°—JL Ш v ¹ F νη ₂ ν-Χν η°—L if ^Ηα F nh ₂ N^X _n ^h °x X j F nh ₂ nXn ^h °—JL if v ¹ F νη ₂ ⁿ °xxY Yes nh ₂ N -Xn ^H °—JL if N^^O^J nh ₂ N^N ^h °—Jl if and a pharmaceutically acceptable salt thereof. - 38 039955 2. A pharmaceutical composition for the treatment of a disorder involving toll-like receptor 7 (TLR7) modulation, comprising a compound of formula according to claim 1, or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable excipients, diluents, or carriers. 3. The use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof as a drug which is a TLR7 modulator. 4. Application of the connection by π. 1 or a pharmaceutically acceptable salt thereof in the treatment of a disorder in which TLR7 modulation is involved. 5. Use of a compound according to claim 4, wherein the disorder is a viral infection. ^jgj) Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky per., 2