EA038093B1 - METHOD OF TREATING A DISEASE CAUSED BY A MYCOBACTERIUM TUBERCULOSIS COMPLEX INFECTION USING (S)-(3-CHLORO-7,8-DIHYDRO-2H-1,6,9-TRIOXA-9a-BORABENZO[cd]AZULEN-2-YL)METHANAMINE OR A PHARMACEUTICALLY ACCEPTABLE SALT THEREOF - Google Patents

Abstract

Provided is a method of treating a disease caused by a Mycobacterium tuberculosis complex infection in an animal comprising: administering to the animal in need of such treatment an effective amount of a compound which is (S)-(3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo[cd]azulen-2-yl)methanamine or a pharmaceutically acceptable salt thereof.

Description

A priority

This application takes precedence of US Provisional Application No. 61/864496, filed August 9, 2013, and US Provisional Application No. 61/918976, filed December 20, 2013, the entire contents of both applications are incorporated herein by reference.

Scope of invention

This invention relates to a method for treating a disease caused by an infection of the Mycobacterium tuberculosis complex in an animal, comprising administering to an animal in need of such treatment an effective amount of a compound representing (S) - (3-chloro-7,8-dihydro-2H1,6 , 9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or a pharmaceutically acceptable salt thereof.

Prior art

Mycobacteria are a genus in a class of bacteria called Actinobacteria and belong to their own family known as Micobacteriacae. Mycobacteria contain a variety of obligate and opportunistic animal pathogens that can also be transmitted to humans and cause disease in humans, thus exhibiting significant zoonotic potential Over the past several decades, members of the Mycobacterium avium-intracellulare complex (MAIC) have emerged as pathogens for human diseases, including lymphadenitis in children, pulmonary tuberculosis-like disease and disseminated infections (occurring mainly in people with weakened immunity, in particular in patients with AIDS). Likewise, important diseases in animals are the result of infection of the animal by members of this group, for example avian tuberculosis and paratuberculosis in ruminants. MAIC includes M. intracellulare and 4 subspecies of M. avium, namely M. avium subsp. avium, M. avium subsp. hommissuis, M. avium subsp. silvaticum and M. avium subsp. paratuberculosis. While members of the M. tuberculosis complex are transmitted by direct contact with the host, MAIC species are acquired predominantly from environmental sources, including soil, water, dust, and food.

Mycobacterium tuberculosis (MTB) is a small, aerobic, immobile rod, high in guanine and cytosine (high-GC), with an outer membrane that is unusually thick, waxy, hydrophobic, rich in mycolic acids and extremely impermeable, making it difficult to treat mycobacterial infections. One third of the world's population is considered infected (including latent MTB), but this number rises to 80% of the population in many countries in Asia and Africa. Without treatment, mortality from active MTV infections is over 50%. In addition, the combination of HIV and MTB is lethal, and an increasing number of MTB strains are becoming resistant to standard drugs, with an estimated 300,000 new cases of multidrug resistance (MDR) of M. tuberculosis every year. Multidrug-resistant (MDR) M. tuberculosis is resistant to isoniazid and rifampicin, and extensively drug-resistant (XDR) M. tuberculosis is also resistant to at least one quinolone and one aminoglycoside. As can be seen in FIG. 1, XDR M. tuberculosis has been recorded over most of the world.

Add to these problems the ease of transmission, as shown in FIG. 2, the globalization of travel and the continuous movement and emigration of many segments of the world's population, it becomes apparent that MTV is becoming a global critical situation.

Synthetic drugs for the treatment of tuberculosis (TB) have been available for over half a century, but the number of cases of the disease continues to rise worldwide. Over 2 billion people are currently infected with M. tuberculosis, most of them latent cases, and it is estimated that over 9 million new cases occur worldwide each year, resulting in between 1.7 and almost 2 million deaths per year. In 2004 alone, approximately 24,500 new infections and almost 5,500 deaths were reported daily (see Zignol, Met al., M. Surveillance of antituberculosis drug resistance in the world an updated analysis, 2007-2010. Bull World Health Organ 2012, 90 (2) , 111-119D). Coinfection with HIV contributes to an increase in morbidity (Williams, BG, Dye, C. Science, 2003, 301, 1535) and the cause of death in 31% of AIDS patients in Africa can be attributed to TB (see Corbett, EL et al., Arch. Intl. Med., 2003, 163, 1009, Septkowitz, A. et al., Clin. Microbiol. Rev. 1995, 8, 180).

The limitations of therapy and prevention of tuberculosis are well known. The current BCG vaccine was released in 1921 and fails to protect most people beyond childhood. According to the 2006 report International Standards for Tuberculosis Care, a document developed by the Tuberculosis Coalition for Technical Assistance (TBCTA), which partners with the Centers for Disease Control, Thoracic Society of America, Tuberculosis Foundation, KNCV, World Health Organization and International Union against Tuberculosis and Lung Disease, infected patients with active disease are currently undergoing a two-month combination therapy with drugs introduced 50-60 years ago, i.e. isoniazid (1952), rifampin (1963), pyrazinamide (1954) and ethambutol (1961), followed by another 4 months of therapy with isoniazid and rifampin (also known as rifampicin). Alternatively, the continuation phase could include isoniazid and ethambutol for six months when adherence cannot be assessed but agreement

- 1 038093 but in this report, a longer continuation phase is associated with a higher rate of relapse and failure, especially in patients with HIV infection. Moreover, as detailed in this report, the doses of anti-tuberculosis drugs used must comply with international guidelines, and fixed dose combinations of two (isoniazid and rifampicin), three (isoniazid, rifampicin and pyrazinamide) and four (isoniazid, rifampicin, pyrazinamide and ethambutol) drugs, especially when it is impossible to monitor patient compliance with the treatment regimen.

Daily dosing is required during these phases of treatment, and poor adherence to treatment leads to the emergence and proliferation of multidrug-resistant strains that are problematic for treatment. Shorter courses of treatment with more active agents, which can be taken less frequently and which create a barrier to the emergence of resistance, i. E. agents that are effective against multi-drug resistant TB (MDRTB) strains. A March 2013 posting (http://www.AIDSmap.com/Once-weekly-continuation-phase-TBtreatment-equals-standard-of-care/page/2589498/) states that a combination of two drugs rifapentine ( a long-acting rifampicin derivative) with moxifloxacin (a fluoroquinolone antibiotic not previously used in TB treatment) can provide once-weekly treatment of TB (TB) during the four-month continuation phase and achieve the same treatment standard as continued treatment with a daily intake of isoniazid and rifampin. This phase of treatment can provide the ability to monitor treatment throughout the continuation phase, which increases adherence to treatment. However, moxifloxacin has not yet received approval for the treatment of TB, and the once-weekly treatment protocol has not yet been endorsed or approved as an alternative treatment standard. International and national peer review groups will need to review published evidence to determine whether this continuation protocol should be recommended and accepted. In addition, rifapentine is expensive, and interactions between rifapentine and antiretroviral drugs in the non-nucleoside reverse transcriptase inhibitor (NNRTI) and protease inhibitor classes may interfere with its use in TB patients who are also HIV positive and are taking antiretroviral drugs. Thus, the cost / benefit analysis of continued treatment with weekly rifapentine versus daily rifampicin is not fully understood at this time.

The anti-TB drug Sirturo ™ (bedaquiline) was approved and approved in the United States at the end of December 2012, and another drug, delamanid, is seeking regulatory approval in the EU. However, both are for drug-resistant tuberculosis, which accounts for only 5% of new cases. The 2007 Editorial and News Focus section of Nature Medicine discusses many aspects of TB, such as pathogenesis, epidemiology, drug discovery and current vaccine development (Nature Medicine, 2007, Focus on Tuberculosis, Vol 13 (3), pages 263 -312), and it is noted that 125 years after the discovery of Mycobacterium tuberculosis, more than one third of the world's people are infected with M. tuberculosis, and of these, more than 1 in 10 will develop a disease known as tuberculosis, formerly known as consumption, in their lifetime.

With the emergence of multidrug-resistant strains of Mycobacterium tuberculosis (MDR-TB), the problem is increasing. The global growth of bacteria and other microorganisms resistant to antibiotics and antibacterial agents in general poses a serious threat. The introduction of a huge number of antimicrobial agents into the ecosphere over the past 60 years has created powerful selective prerequisites for the emergence and spread of antimicrobial-resistant pathogens. Therefore, there is a need for the discovery and development of new chemicals for TB treatment (recent leaders reviewed in Grosset JH, Singer TG, Bishai WR New Drugs for the Treatment of Tuberculosis Hope and Reality Int J Tuberc Lung Dis 2012 Aug, 16 (8) 1005 -fourteen).

This document discloses tricyclic benzoxaborol compounds that exhibit unexpected selectivity for inhibiting replication of Mycobacterium tuberculosis (M. tuberculosis) in relation to inhibition (toxicity) of human cells compared to other benzoxaborol compounds and exhibiting submicromolar MIC values against mycobacterium species, in particular Mycobacterium tuberculosis and Mycobacterium tuberculosis complex (MTC), Mycobacterium avium and Mycobacterium avium complex (MAC) and Mycobacterium avium intracellulare complex (MAIC). Generally speaking, benzoxaborole has the following structure and numbering system for substituents:

he /

Some benzoxaborols that are substituted at the 7-position form a tricyclic compound

- 2 038093 benzoxaborol. When the resulting tricyclic benzoxaborole is further substituted with a halogen substituent at position 4 and an aminomethyl substituent at position 3, then such compounds are surprisingly selective for mycobacteria and effective against mycobacteria, including M. tuberculosis. The observed selectivity is assessed by comparing the MIC (Minimum Inhibitory Concentration) values for such compounds with respect to the inhibitory effect (toxic effect) of these compounds on human cells compared to other benzoxaborol compounds.

Boron-containing molecules such as benzoxaboroles, which are useful as antimicrobial agents, have been described previously (see, for example, Benzoxaboroles - Old compounds with new applications Adamczyk-Wozniak, A. et al., Journal of Organometallic Chemistry Volume 694, Issue 22, 15 October 2009, Pages 3533-3541, and US Patent Publications US 20060234981, US 20070155699, US 20090227541, WO 2012033858 and US 2013165411.

US20090227541 describes numerous compounds, including two tricyclic benzoxaborol compounds with distinct antibacterial activity against a panel of gram-negative bacteria (see, eg, Tables 1 and 2), but does not disclose tricyclic benzoxaborol compounds with halogen substitution on the benzoxaborole ring. WO 2012033858 discloses benzoxaborol compounds with activity against Mycobacterium tuberculosis, including some benzoxaborol compounds (see, for example, examples 1.A-1.Y), but again no tricyclic benzoxaborol compounds with halogen substitution on the benzoxaborole ring are disclosed. US 2013165411 discloses tricyclic benzoxaborol compounds showing activity against Acinetobacter baumannu, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae (see Table 1), but specifically states that the investigated halogen-substituted tricyclic compounds (examples 17, 18 and 19) have no activity against A. baumannii, having antibacterial activity with MIC values> 16 μg / μl (see Fig. 1B).

Summary of the invention

The present invention provides a method of treating a disease caused by an infection of the Mycobacterium tuberculosis complex in an animal, comprising administering to an animal in need of such treatment an effective amount of a compound representing (S) - (3-chloro-7,8-dihydro2H-1,6 , 9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or a pharmaceutically acceptable salt thereof, wherein said salt is preferably ((S) - (3-chloro-7,8-dihydro-2H-1 , 6,9-trioxa-9aborabenzo [cd] azulen-2-yl) methanamine dihydrosulfate-H ₂ O.

In one aspect of the invention, said compound is administered as a pharmaceutical composition with at least one pharmaceutically acceptable excipient.

In yet another aspect of the invention, said compound is administered in combination with a second therapeutic agent that is not the compound or a pharmaceutically acceptable salt thereof as defined above.

Preferably, said combination comprises a pharmaceutically acceptable excipient, adjuvant, or diluent.

In a preferred embodiment of the method according to the invention, the second therapeutic agent is independently selected from isoniazid, rifampin, pyrazinamide, ethambutol, moxifloxacin, rifapentin, clofazimine, bedaquiline (TMC207), nitroimidazooxazine PA-824, delamanid (OPC-67683), an analogue of ethazolautinone SQ109, benzothiazinone and dinitrobenzamide.

Preferably, the oxazolidinone is linezolid, tedizolid, radezolid, sutezolid (PNU-100480) or posizolid (AZD-5847).

In another preferred embodiment of the method of the invention, the second therapeutic agent is an antiviral agent.

Preferably, said antiviral agent is an antiretroviral agent.

Even more preferably, the anti-retroviral agent is zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxil, fozivudine, todoxil, emtricitabine, alovudine, amdoxovir, elvucitabine, delavirziviroval, efvirpravine , GSK2248761, TMS-278, TMS-125, etravirine, saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, brekanavir, darunavir, atazanavir, tipranavir, palinavir, lasinavirtide-2, enfu-12 , PRO-140, TNX-355, BMS-806, BMS-663068 and BMS-626529, 5-Helix, raltegravir, elvitegravir, GSK1349572, GSK1265744, vikrivirok (Sch-C), Sch-D, TAK779, maravirok, TAK449, didanosine, tenofovir, lopinavir, or darunavir.

In another preferred embodiment of the method of the invention, the second therapeutic agent is a therapeutic agent approved or recommended for the treatment of tuberculosis.

In another aspect of the invention, said disease is tuberculosis.

In another aspect of the invention, said animal is a human.

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Brief description of graphic materials

FIG. 1 is a world map showing where XDR-TB is geographically documented.

FIG. 2 shows the transmission of tuberculosis.

FIG. 3 is a graph of MIC values (from Tables 1A and 1B) for the compound of Example 4 G4Cl versus clinical isolates of M. tuberculosis.

FIG. 4 is a graph of MIC (from Tables 2A, 2B, 2C and 2D) for the compound of Example 2 and the compound of Example 4 (G2-Br and G4-C1, respectively) against clinical isolates of M. tuberculosis.

Table 1A and 1B show the MIC values for Example 4 compound G4-Cl tested against 97 clinical isolates of M. tuberculosis: susceptible (A) and resistant (B). Table 1A shows the MIC results for the compound of Example 4 in relation to M. tuberculosis strains susceptible to known anti-TB agents, and in table. 1B shows the MIC results for the compound of Example 4 against M. tuberculosis strains resistant to one or more known anti-TB agents. The resistance pattern of clinical isolates is shown by the following abbreviations: H isoniazid, R is rifampicin, T is ethionamide, S is streptomycin, E is ethambutol, Z is pyrazinamide, K is kanamycin, A is amikacin, and CP is capreomycin.

Table 2A and 2B show the MIC values for the compound of Example 4 G4-Cl tested against 40 strains of clinical isolates of M. tuberculosis: susceptible (A) and resistant (B). Table 2A shows the MIC results for the compound of Example 4 in relation to M. Tuberculosis strains susceptible to (Standard for TB Treatment Agents), and in Table. 2B shows the MIC results for the compound of Example 4 against M. tuberculosis strains resistant to one or more known anti-TB agents.

Table 2C and 2D show the MIC values of Example 2 compound G2-Br tested against 40 M. tuberculosis strains. Clinical isolates: sensitive (A) and resistant (B). Table 2C shows the MIC results for the compound of Example 2 in relation to M. Tuberculosis strains susceptible to known anti-TB agents, and in table. 2D shows the MIC results for the compound of Example 2 against M. tuberculosis strains resistant to one or more known anti-TB agents. The resistance pattern of clinical isolates is shown by the following abbreviations: H - isoniazid, R - rifampicin, T - ethionamide, S - streptomycin, E - ethambutol, Z - pyrazinamide, K - kanamycin, A - amikacin, and CP - capreomycin.

Table 3 shows the MIC values with respect to non-mycobacterial strains for compounds of formula II or formula IIa.

Table 4A shows the _{LeuRS inhibition IC 50} values, the MIC values relative to the standard M. tuberculosis Mtb H37Rv strain, the toxicity values to human HepG2 cells, and the selectivity values for several comparative benzoxaborol tricyclic compounds.

Table 4B presents the classification of the data shown in table. 4A, for compounds of Formula II or Formula IIa.

Detailed description of specific embodiments

This document discloses a tricyclic benzoxaborole compound or a salt thereof having the structure of Formula II

Formula II wherein X is selected from chlorine, fluorine, bromine and iodine, each of R ¹ and R ^{2 is} independently selected from H, —CH ₃ , —CH2CH3, —CH2CH2CH3, and —CH (CH3) 2;

and also a compound of formula IIa

Formula Pa where X is fluoro, chloro, bromo or iodo and R ¹ and R ^{2 are} independently H or —CH 3, or a pharmaceutically acceptable salt thereof.

- 4 038093

The compound used in the process according to the invention, which is (S) - (3-chloro-7,8dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, has a structure corresponding to the indicated structures of formulas II and IIa.

The inventors have surprisingly found that the tricyclic benzoxaborol compounds described herein exhibit an unexpected replication inhibition selectivity of Mycobacterium tuberculosis (M. tuberculosis) relative to the inhibitory effect (toxic effect) on human cells over other benzoxaborol compounds These tricyclic benzoxaborol compounds exhibit submicromolecular values MICs against M. tuberculosis that are comparable to or better than MIC values for current therapies available to inhibit M. tuberculosis. In addition, in other embodiments, the benzoxaborol tricyclic compounds described herein are intended for use in combination with current anti-tuberculosis compounds and are envisioned to be more effective in treating animals, including humans, infected with M. tuberculosis.

Resistance remains an issue in the treatment of tuberculosis (TB), and one clinical strategy is to focus on timely combination with other TB drugs and expedite the timely evaluation of the compounds' efficacy in patients. Compounds of Formula II or Formula IIa offer a unique opportunity to address serious problems that arise during TB treatment, such as multidrug resistance, extensive drug resistance, reactivity and / or deleterious interactions between therapeutic agents in a multidrug combination and duration of treatment, thereby satisfying the needs of potential patients.

Animal in this document means any of the kingdom (Animalia) of living things, including multicellular organisms, including livestock and pets, including cattle, sheep, goats, dogs and cats, or a person, including a person with a weakened immune system.

A combination of the invention, as used herein, refers to combinations of the compounds contemplated herein, salts (eg, pharmaceutically acceptable salts), prodrugs, solvates, and hydrates of these compounds.

A diastereomer, as used herein, refers to one of a pair of stereoisomers that is not a mirror image of the other stereoisomer.

An enantiomer, as used herein, refers to one of a pair of non-superimposed racemic compounds (racemates) that is a mirror image of the other enantiomer. Enantiomers in pure form have the property of rotating the plane of polarized light in one direction or in the other direction, but in the form of a racemic mixture, the mixture does not rotate the plane of polarized light.

An effective amount of a compound, a combination thereof or a composition thereof means the amount of a compound that is an active agent, including a combination or composition thereof, that is, an amount sufficient to provide the desired local or systemic effect. A therapeutically effective or pharmaceutically effective amount refers to an amount of a compound, including a combination or composition thereof, sufficient to achieve the desired therapeutic or pharmaceutical result.

The term pharmaceutically acceptable salt encompasses a salt of a compound described herein that is prepared with relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein. When the compounds described herein contain relatively acidic functional groups, then base addition salts can be prepared by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium salt, organic amine addition salt (such as choline or diethylamine, or amino acids such as d-arginine, l-arginine, d-lysine or l-lysine) or magnesium salt, or similar salt. When the compounds described herein contain relatively basic functional groups, then acid addition salts can be prepared by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts formed with inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrocarbon, phosphoric, monohydrophosphoric, dihydrophosphoric, sulfuric, monohydrosulfuric, hydroiodic or phosphorous acid, and the like, as well as salts, formed with relatively non-toxic organic acids such as acetic, propionic, isobutyric, maleic, malic, benzoic, succinic, cork, fumaric, lactic, almond, phthalic, benzenesulfonic, paratolylsulfonic, citric, tartaric, methanesulfonic, etc. Also encompassed are salts with amino acids such as arginate and the like, and salts with organic acids such as glucuronic or galacturonic acid and the like. (see, for example, Berge et al., Pharmaceutical Salts, Journal of Pharmaceutical Science 66 1-19 (1977)).

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Certain specific compounds described herein contain both basic and acidic functional groups, which make it possible to convert the compounds to either base addition salts or acid addition salts.

The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compounds in a conventional manner. The parent form of the compound differs from the various salt forms in some physical properties, such as solubility in polar solvents.

In addition to salt forms, the disclosed compounds may be in the form of prodrugs. Prodrugs of the compounds described herein readily undergo chemical transformations under physiological conditions to form the compounds described herein. Additionally, prodrugs can be converted to compounds of the invention by chemical or biochemical methods ex vivo.

Certain compounds of Formula II and Formula IIa may form acid addition salts with one or more acid equivalents.

The compounds of formula II and formula IIa can be obtained in crystalline or non-crystalline form and, if crystalline, can optionally be solvated, for example in the form of a hydrate. They can be stoichiometric solvates (eg hydrates) as well as compounds containing variable amounts of solvent (eg water). Also contemplated are isotopically labeled compounds that are identical to the compounds of Formula II and Formula IIa, but differ in that one or more atoms are replaced by an atom having an atomic mass or mass number other than the atomic mass or mass number normally found in nature. Examples of isotopes that can be introduced into the compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine, and chlorine, such as ³ H, C. ¹⁴ C, ¹⁸ F, ¹²³ I, or ¹²⁵ I.

The disclosed isotopically labeled compounds, for example compounds into which radioactive isotopes such as ³ H or ¹⁴ C have been incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, i.e. ³ H, and carbon-14, i.e. ¹⁴ C isotopes are particularly preferred for their ease of preparation and detection. The isotopes ^P C and ¹⁸ F are especially useful in PET (Positron Emission Tomography).

Since the compounds of Formula II and Formula IIa described herein are intended for use in pharmaceutical compositions, it will be understood that each of them is preferably provided in substantially pure form, for example, with a purity of at least 60 wt%, preferably with a purity of at least 75 wt.% and preferably with a purity of at least 85 wt.%, especially with a purity of at least 98 wt.%. Formulations of compounds with impurities can be used to obtain purer forms used in pharmaceutical compositions.

In one specific embodiment, treatment of a mycobacterial infection or condition occurs through inhibition of an aminoacyl tRNA synthetase editing domain by binding to an editing active site. In yet another illustrative embodiment, treatment of a mycobacterial infection or condition occurs by blocking an aminoacyl tRNA synthetase editing domain.

Pharmaceutical compositions.

Information regarding excipients for use in a pharmaceutical composition for use in a method according to the invention can be found in Remington The Science and Practice of Pharmacy, 21st Ed., Pharmaceutical Press (2011), which is incorporated herein by reference.

Combinations.

In an illustrative embodiment of the invention, the benzoxaborol tricyclic compound is (S) - (3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or a pharmaceutically acceptable salt thereof, as described herein, is administered in combination with a second therapeutic agent. In some embodiments, the second therapeutic agent is an antibacterial agent, more specifically an anti-tuberculosis agent, more specifically an anti-M. tuberculosis agent.

In an illustrative embodiment, the combination is part of a pharmaceutical composition described herein. Such circumstances are known to a person skilled in the art, and specific circumstances are set forth in the examples given in this description.

Combination dosage forms.

The individual components of the combinations for use in the method of the invention, for example the combination described herein, can be administered either simultaneously or sequentially in unit dosage form. The unit dosage form can be a single or multiple unit dosage form. In an illustrative embodiment of the invention, a combination is provided in a single unit dosage form. An example of a single unit dosage form is a capsule where the tricyclic benzoxaborol compound is (S) - (3-chloro-7,8-dihydro-2H-1,6,9-mrioxa-9a-borabenzo [cd] azulene-2- sludge) methane

- 6 038093 amine, or a pharmaceutically acceptable salt thereof, and the additional therapeutic agent are both contained within the same capsule. In an illustrative embodiment of the invention, a combination is provided in a dual unit dosage form. An example of a double unit dosage form is a first capsule that contains a benzoxaborol tricyclic compound, which is (S) - (3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene- 2-yl) methanamine, or a pharmaceutically acceptable salt thereof, and a second capsule that contains an additional therapeutic agent. Thus, the term single unit, or double unit, multiple unit refers to the object being swallowed by the patient and not to the internal components of the object. Suitable dosages of said tricyclic benzoxaborol compound will be readily determined by those skilled in the art. Suitable dosages of an additional therapeutic agent that is not a compound of Formula II or Formula IIa will be readily determined by those skilled in the art. In one specific embodiment, said tricyclic benzoxaborol compound is present in combination in a therapeutically effective amount. In one specific embodiment, the additional therapeutic agent, which is not a compound of Formula II or Formula IIa, is present in combination in an amount sufficient to kill or reduce the presence, amount, or growth rate of mycobacteria, including M. tuberculosis, by the substituted benzoxaborol.

Additional therapeutic agent (s) in combination.

The claimed method provides, in one aspect, the introduction of a combination containing a tricyclic benzoxaborol compound, which is (S) - (3-chloro-7,8-dihydro-2H-1,6,9-trioxa9a-borabenzo [cd] azulen-2-yl ) methanamine described herein, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent. In an illustrative embodiment, the additional therapeutic agent is an antimycobacterial agent.

The combinations may conveniently be presented for use in the form of a pharmaceutical composition. The individual components of such combinations can be administered either simultaneously or sequentially in separate pharmaceutical compositions or in a combined pharmaceutical composition in a convenient manner.

When an additional therapeutic agent is used with the combination described herein against the same disease state, then the dose of each compound may differ from the dose when the compound is used alone. Suitable dosages will be apparent to those skilled in the art. It will be apparent that the amount of a compound described herein required for use in treatment will vary depending on the nature of the condition being treated and the age and condition of the patient and will ultimately be at the discretion of the treating physician or veterinarian.

Obtaining boron-containing compounds.

The compounds disclosed herein can be prepared using commercially available starting materials, known intermediates, or using the synthetic methods described herein or published in sources of information indicated and included in this description by reference, such as US 7816344 and PCT Patent Publications WO 2010080558 and WO 2011127143. General procedures used to synthesize compounds of Formula II and Formula IIa are depicted in Reaction Schemes 1-5 and illustrated by examples.

Some tricyclic benzoxaborol compounds can be obtained by the Mitsunobu reaction to the conversion of the hydroxyl substituent 2-bromo-3-hydroxybenzaldehyde to tetrahydropyranyloxyethoxyether by reaction with tetrahydropyranyloxyethanol in triphenylphosphine (PPh ₃ ), tetrahydrofuranylation (THF) Miyyaura substituent is bromo, using bis (pinokolato) diboron diboron (B ₂ pin ₂₎ with a palladium catalyst (PdCl _2), and potassium acetate (KOAc), and subsequent reductive cyclization to form a tricyclic compound with sodium borohydride (NaBH ₄ ) in anhydrous methanol (MeOH), as shown in Scheme 1

Scheme 1

OTNR OTNR

Reaction fj \ Borylation / h I /

Mitsunobu Cr2 according to Miyaura θ 2θ X /

ТНРО (СН ₂ ) рн 'ργ ^ΒΓ _B2 pin ₂ ' ργ ^в '° ^^ sno ^^ sno

Reductive θ / ^- \ opening of the ring ι О -

THP-protected alkyl bromide can also be used to add a substituent to hydroxybenzaldehyde via SN2 reaction to give tricyclic benzoxaborol compounds. Examples of using the SN2 reaction to prepare tricyclic benzoxaborol compounds are given in the examples described below, such as example 1, step (b), and example 4, method B, step (c).

Other tricyclic benzoxaborol compounds described in this document may be

- 7 038093 obtained as shown in schemes 2 and 3, where the nitroaldol reaction is carried out on the substituent of the aldehyde, for example 3- (2-benzyloxy-ethoxy) -2- (4,4,5,5-tetramethyl- [1,3 , 2] dioxaborolan-2-yl) benzaldehyde, using nitromethane (MeNO ₂ ) with a base (NaOH) to obtain a nitro-substituted benzyl-substituted benzoxaborole compound, followed by cyclization and reduction of the nitro substituent to an amine using Pd (OH) ₂ / C in ice-cold acetic acid acid to form the target tricyclic benzoxaborol compound

Scheme 2

Scheme 3

Other tricyclic benzoxaborol compounds described herein can be prepared as shown in Scheme 4

Scheme 4

Other tricyclic benzoxaborol compounds described herein can be prepared as shown in Scheme 5

Scheme 5

intermediate connection

Alternatively, some tricyclic benzoxaborol compounds can be prepared as shown in Scheme 6.

A mixture of (S) -tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (13.25 kg) and NCS (8.75 kg) in dichloroethane (132.5 L) was heated at 70 ° C until the reaction was complete by HPLC. The mixture was concentrated under reduced pressure, cooled to 25 ° C and acetone (106 L) added. The suspension was filtered, rinsing with acetone (26.5 L). The wet filter cake was suspended in water (13.25 L) and 1,4-dioxane (66.25 L), heated to 50 ° C for 20-30 min, cooled to 15 ° C, filtered and the filter cake was washed 1,4-dioxane (26.5 L). The wet filter cake was dissolved in methanol (68.9 L), filtered, and the filtrate was concentrated under reduced pressure. Methyl tert-butyl ether (66.25 L) was added to the residue, and the mixture was concentrated under reduced pressure. Methyl tert-butyl ether (78.7 L), isopropanol (8.7 L) and sulfuric acid (4.6 L) were added, the mixture was heated to 50 ° C and stirred until the sulfate content reached 24.32-29 , 72%. The mixture was cooled to 25 ° C, stirred for 1 h, filtered, the filter cake was washed with methyl tert-butyl ether (17.5 L) and dried to give the desired product (42%).

- 8 038093

Scheme 6

As can be seen in Scheme 6, in this synthetic route, the 10/11 and 12/13 end-steps are replaced with alternative end-steps, excluding the deprotection / deprotection steps.

Composition and preparations.

The compounds described herein can be prepared for administration by any convenient route for human or veterinary use in analogy with the preparation of antimycobacterial agents or the preparation of other anti-tuberculosis agents.

Before administration to a patient, a compound that is (S) - (3-chloro-7,8-dihydro-2H-1,6,9trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or a pharmaceutically acceptable salt thereof, described herein, normally, but not necessarily, will be prepared in the form of pharmaceutical compositions, which may also contain one or more pharmaceutically acceptable carriers, excipients or diluents. The carrier, excipient or diluent must be acceptable in the sense of being compatible with the other ingredients of the composition and not hazardous to the recipient.

The pharmaceutical compositions described herein include compositions in a form suitable for oral or parenteral administration and can be used to treat mycobacterial infection in a mammal, including a human.

The pharmaceutical compositions described herein include compositions in a form suitable for oral, topical or parenteral administration and can be used to treat mycobacterial infection in a mammal, including a human.

The composition can be prepared for administration by any convenient route. For the treatment of tuberculosis, the compositions can be in the form of tablets, capsules, powders, granules, troches, aerosols, or liquid preparations such as oral or sterile parenteral solutions or suspensions.

Tablets and capsules for oral administration may be presented in unit dosage form and may contain conventional excipients such as binding agents such as syrup, gum arabic, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone, excipients such as lactose, sugar, maize starch, calcium phosphate , sorbitol or glycine, tabletting lubricants such as magnesium stearate, talc, polyethylene glycol or silica, disintegrants such as potato starch, or acceptable wetting agents such as sodium lauryl sulfate. Tablets can be coated by methods known in conventional pharmaceutical practice. Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a product for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example sorbitol, methylcellulose, glucose syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats, emulsifiers, for example lecithin, sorbitan monooleate, or which gum arabic may include edible oils), for example almond oil, oily esters such as glycerin, propylene glycol or ethyl alcohol, preservatives, for example methyl or propyl parahydroxybenzoate or sorbic acid, and, if desired, traditional flavoring or coloring agents.

Suppositories will contain conventional suppository bases such as cocoa butter or other glyceride.

For parenteral administration, liquid unit dosage forms are prepared using

9 038093 pricing the compound and sterile vehicle, with water being preferred. The compound, depending on the vehicle used and the concentration used, can be either suspended or dissolved in the vehicle. In preparing solutions, the compound can be dissolved in water for injection and sterilized by filtration, then filled into a suitable vial or ampoule and sealed.

In one aspect of the invention, agents such as local anesthetics, preservatives, and buffering agents can be dissolved in the carrier. To enhance stability, the composition can be frozen after filling into a vessel and the water removed under vacuum. The dry lyophilized powder is then sealed in a vessel, and an accompanying vessel of water for injection can be supplied to reconstitute the powder prior to use. Parenteral suspensions are prepared in substantially the same manner, except that the compound is not dissolved but suspended in a vehicle, and sterilization is optionally accomplished by filtration. The compound can be sterilized with ethylene oxide before being suspended in a sterile vehicle. Advantageously, a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the compound.

The compositions may contain from 0.1% by weight, preferably from 10 to 60% by weight, of active substance, depending on the route of administration. If the compositions contain dosage units, then each unit will preferably contain from 20 to 1000 mg of the active ingredient. The dosage used to treat an adult will usually be in the range of 50 to 300 mg per day, for example, 150 to 200 mg per day, depending on the route and frequency of administration. This dosage corresponds to 0.5-5 mg / kg per day. A preferred dosage is from 0.5 to 2 mg / kg per day, and a more preferred dose is less than 1 mg / kg per day.

A compound of formula II or formula IIa, which is (S) - (3-chloro-7,8-dihydro-2H-1,6,9trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or a pharmaceutically acceptable the salt may be the only therapeutic agent in the compositions described herein, or may be present in the composition in combination with one or more additional therapeutic agents.

The one or more additional therapeutic agents is, for example, an agent useful for treating tuberculosis in a mammal. Examples of such therapeutic agents include rifampin, pyrazinamide, ethambutol, moxifloxacin, rifapentine, clofazimine, bedaquiline (TMC207), nitroimidazooxazine PA-824, delamanid (ORS-67683), oxazolidinone such as linezolid, radezolidizolid, PNU-100 and posizolid (AZD-5847), an EMB SQ109 analogue, benzothiazinone, dinitrobenzamide and antiviral agent, including an antiretroviral agent, or any anti-TB agent in development for the treatment of TB, with a positive response in phase IIa EVA trials, or any agent against TB developed by the Global Alliance Against Tuberculosis.

When a compound of formula II or formula IIa, which is (S) - (3-chloro-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or pharmaceutically an acceptable salt is used in combination with one or more additional therapeutic agents, then the dose of the compound or agent may differ from the dose when only the compound or only the agent is used. Suitable dosages will be readily determined by those skilled in the art. It will be understood that the amount of a compound described herein and one or more additional therapeutic agents required for use in treatment will vary depending on the nature of the condition being treated and the age and condition of the patient and will ultimately be at the discretion of the attending physician or veterinarian.

The combinations may conveniently be presented for use in the form of a pharmaceutical composition. The combination may contain a compound of formula II which is (S) - (3-chloro7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or a pharmaceutically acceptable a salt together with one or more additional therapeutic agents and one or more pharmaceutically acceptable carriers, excipients or diluents. The individual components of such combinations can be administered either sequentially or simultaneously in separate pharmaceutical compositions or combined pharmaceutical compositions in any convenient way.

When the administration is sequential, either the compound being (S) - (3-chloro7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine, or pharmaceutically an acceptable salt, or one or more additional therapeutic agents may be administered first. When the administration is simultaneous, the combination can be administered either in the same pharmaceutical composition or in different pharmaceutical compositions. Obviously, when combined in the same composition, the compound and agents must be stable and compatible with each other and with other components of the composition. When prepared separately, they can be provided in a convenient composition in a manner known to such compounds in the art.

Abbreviations.

In the description of the invention, chemical elements are identified in accordance with the Periodic Table of the Elements. Abbreviations and symbols used in this document correspond to general

- 10 038093 accepted abbreviations and symbols used by specialists in chemical fields. The following abbreviations are used:

Ason acetic acid Ac ₂ O AIBN acetic anhydride 2-2'-azobutyronitrile VOS N-Crack Butoxycarbonyl BOC anhydride B ₂ pin ₂ di-tert-butyl dicarbonate bis (pinacolato) dibor dibor, also known as 4,4,4 ', 4', 5,5,5 ', 5'octamethyl-2,2'-bi (1,3,2-dioxaborolane) Celite® filter bed composed of acid washed diatomaceous silicon dioxide (trademark of Manville Corp, Denver, Colorado) STAV cetyltrimethylammonium bromide DCM dichloromethane DIAD diisopropyl azodicarboxylate DIBAL-H diisobutylaluminum hydride DME dimethoxyethane DCE dichloroethane DMF dimethylformamide DMSO-d6 deuterated dimethyl sulfoxide DMSO dimethyl sulfoxide ERI electrospray ionization ERI MC spray ionization mass spectrometry Et ₂ O diethyl ether EtOH ethanol EtOAc, EA ethyl acetate 4 watch HPLC high performance liquid chromatography KOAc potassium acetate LC / MS liquid chromatography / mass spectroscopy mCPBA meta-chloroperbenzoic acid MeNO ₂ nitromethane MeOH methanol NBS N-bromosuccinimide NCS NIS N-chlorosuccinimide N-iodosuccinimide NXS N-halosuccinimide NaBH (OAc) ₃ NMR sodium triacetoxyborohydride nuclear magnetic resonance spectroscopy PE petroleum ether PPh ₃ triphenylphosphine kt or kt room temperature RT retention time SFC supercritical fluid chromatography t-BuOMe methyl-ot-butyl ether TFA trifluoroacetic acid THF tetrahydrofuran Uv ultraviolet radiation

Examples of

The examples below illustrate the invention. These examples are not intended to limit the scope of the invention, but provide guidance to those skilled in the art in the preparation and use of the compounds, compositions and methods of the invention. While specific embodiments of the invention have been described, one skilled in the art will appreciate that various changes and modifications can be made. References to preparations made by analogy with other preparations or a general preparation method may cover variations in routine parameters such as time, temperature, operating conditions, minor changes in the amounts of reagents, etc.

Recorded proton nuclear magnetic resonance spectra ( ¹ H NMR), and chemical shifts

- 11 038093 gi are given in parts per million (δ) descending from the internal tetramethylsilane standard (TMS). Abbreviations for NMR data are as follows: s - singlet, d - doublet, t - triplet, q - quartet, m multiplet, dd - doublet of doublets, dt - doublet of triplets, app - visible, br - broadened. Mass spectra were obtained using electrospray ionization (ERI) methods. All temperatures are in degrees Celsius.

Reactions involving metal hydrides, including lithium hydride, lithium aluminum hydride, diisobutyl aluminum hydride, sodium hydride, sodium borohydride, and sodium triacetoxyborohydride, were performed under argon, unless otherwise indicated.

Synthesis.

Example 1 (comparative). 3-Bromo-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (G1-Br)

(a) 2-bromo-3-hydroxybenzaldehyde on he

D. Br ₂ / CH ₃ COONa B J _{/ O} Fe / AcOH

A suspension of 3-hydroxybenzaldehyde (5 g, 40 mmol), iron powder (172 mg, 3 mmol) and sodium acetate (6.72 g, 80 mmol) in acetic acid (40 ml) was heated until a clear solution was obtained and then cooled to room temperature. temperature. To this mixture was added a solution of bromine (7.2 g, 45 mmol) in glacial acetic acid (10 ml) dropwise over 15 minutes. After the addition, the reaction mixture was stirred for 2 h and then poured into ice-water. The resulting mixture was extracted with dichloromethane (3x50 ml). The combined extracts were dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was recrystallized from dichloromethane to give the product (2.3 g, 28%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.75 (s, 1H), 10.26 (s, 1H), 7.38-7.24 (m, 3H).

(b) 2-Bromo-3- (2 - ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) benzaldehyde

OH 1) DHP - ° ^THP

X.Br Br (Jl Q 2) ^K 2 ^C ° 3- KI- L JUo

DMF, 70 ° C

Dihydropyran (1.26 g, 15 mmol) was added dropwise at 0 ° C. to 2-bromoethanol (1.875 g, 15 mmol). The mixture was stirred for 30 min at 0 ° С and then for 2 h at rt. To this mixture was added 2-bromo-3-hydroxy benzaldehyde (2 g, 10 mmol), followed by potassium carbonate (1.518 g, 11 mmol), potassium iodide (332 mg, 2 mmol) and dry DMF (20 ml). The reaction mixture was stirred at 70 ° C overnight. The solution was cooled to rt and diluted with diethyl ether (100 ml). The inorganic salts were removed by filtration and the filtrate was diluted with hexanes (100 ml). The organic layer was washed with water (50 ml x 3) and then concentrated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel using ethyl acetate and petroleum ether as eluants to give the title compound (3 g, 92%) as a yellow oil. MS (ERI) m / z = 351 [M + 23] +, Rf = 0.7 (PE: EA = 3) ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.29 (s, 1H), 7.50- 7.41 (m, 3H), 4.75 (s, 1H), 4.31-4.28 (m, 2H), 4.00-3.94 (m, 1H), 3.82-3.75 (m, 2H), 3.47-3.43 (m, 1H), 1.73-1.50 (m, 6H).

(c) 3 - (2 - ((T tetrahydro-2H-pyran-2-yl) oxy) ethoxy) -2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl ) benzaldehyde _o ^ OTNP THPO ^ x V

T _Br Pin ₂ B ₂ , PdCI ₂ (dppf), ϊ 4 А ί | Υ KOАс, 90 ° С '0 ⁴

A solution of 2-bromo-3- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) benzaldehyde (160 g, 0.49 mol), 4.4.4 ', 4', 5.5.5 ' , 5'-octamethyl-2,2'-bi- (1,3,2-dioxaborolane) (249 g, 0.98 mol), Pd (dppf) Cl ₂ (20 g, 24.5 mmol) and KOAc ( 144 g, 1.47 mol) in DMF (2.0 L) was stirred at 90 ° C overnight. The reaction mixture was then treated with water (4 L) and then extracted with EtOAc (3x1.5 L). The combined organic layers were washed with brine (250 ml), dried over anhydrous Na2SO4 and concentrated to dryness in vacuo. The residue was purified by silica gel column chromatography (petroleum ether ethyl acetate = 10: 1 to 2: 1) to give the title compound as a yellow oil (88 g, 48% yield) MS (ERI) m / z = 317 [M + H] +, Rf = 0.4 (PE: EA = 3) ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 9.88 (s, 1H), 7.60-7.51 (m, 2H), 7.31-7.28 (d, 1H ), 4.64-4.63 (m, 1H), 4.16-4.13 (m, 2H), 4.00-3.94 (m, 1H), 3.82-3.75 (m, 2H), 3.47-3.43 (m, 1H), 1.73-1.50 (m, 6H), 1.29 (m, 12H).

(d) 2- (Nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene

- 12 038093

To a solution of NaOH (4.8 g, 0.12 mol) in water (100 ml) was added nitromethane (18.3 g, 0.3 mol) at 510 ° C. After stirring for 15 min at 5-10 ° C, cetyltrimethylammonium bromide (CTAB) (2.2 g, 6 mmol) was added to the reaction mixture and then 3- (2- (tetrahydro-2H-pyran-2yloxy) ethoxy) - 2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) benzaldehyde (45 g, 0.12 mol) at 510 ° C. The reaction mixture was stirred at rt for 5 h. The reaction mixture was acidified to pH 1 using dilute hydrochloric acid and stirred at rt overnight. The reaction mixture was filtered to give the title compound (14.5 g, 51%) as a white solid. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.50-7.45 (t, 1H), 7.16-7.13 (d, 1H), 6.91-6.88 (d, 1H), 5.91-5.88 (m, 1H), 5.37 -5.31 (m, 1H), 4.69-4.61 (m, 2H), 4.41-4.14 (m, 3H).

(f) (7,8-Dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride

A solution of 2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene (1.5 g, 6.4 mmol), Raney Ni (200 mg) and 2 M NH ₃ in EtOH (5 ml) in ethanol (40 ml) was shaken in an atmosphere of H ₂ for 2 h at rt. The mixture was filtered through a Celite pad and the filtrate was concentrated in vacuo. The crude amine was dissolved in EtOH (20 ml) and a saturated solution of HCl (gas) in Et ₂ O (30 ml) was immediately added. After 1 h, the suspension was filtered and the resulting solid washed with acetonitrile / hexanes (2: 1, 2x20 ml) to give the compound as a white solid (700 mg, 45%) MS (ERI) m / z = 206/224 [M + H] +.

(f) tert-Butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

In a mixture of (7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (700 mg, 2.9 mmol) and triethylamine (878.7 mg, 8.7 mmol) in dichloromethane (10 ml) at 0 ° C was added di-tert-butyl dicarbonate (948 mg, 4.35 mmol) and the mixture was stirred for 2 h at room temperature. The reaction was quenched with saturated NaHCO ₃ (15 ml) and the resulting mixture was extracted with EtOAc (3x20 ml). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by flash chromatography using ethyl acetate and petroleum ether as eluents to give the title product (500 mg, 56%) MS (ERI) m / z = 250 [M-56] ⁺ .

(g) tert-Butyl - ((3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

To a solution of tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate (0.5 mg, 1.64 mmol) and NBS (354 mg, 2.0 mmol) in acetonitrile (15 ml) was added AIBN (27 mg) and the mixture was stirred for 1 h at 90 ° C. The reaction mixture was then concentrated in vacuo and the residue was purified by preparative HPLC to give the desired product (300 mg, 50%). MS (ERI) m / z = 328/330 [M-56] ⁺ .

(h) Header Compound

A mixture of tert-butyl - ((3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (0.2 g, 0.522 mmol ) in a saturated solution of HCl (gas) in Et2O (10 ml) was stirred at rt for 1 h and concentrated to dryness (water bath temperature <30 ° C). The residue was triturated with acetonitrile (2x5 ml) and the white solid was dried under high vacuum to give the product (140 mg, 83%) as a white solid. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 8.36 (s, 3H), 7.64-7.61 (d, 1H), 6.93-6.90 (d, 1H), 5.51-5.49 (d, 1H), 4.69 (m , 1H), 4.36-4.23 (m, 3H), 3.62 (m, 1H), 3.05-3.01 (m, 1H). MS (ERI) m / z = 284/286 [M + H] +.

Example 2 (comparative). (S) - (3-Bromo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2yl) methanamine hydrochloride (G2-Br)

- 13 038093

Method A.

(a) Title compound

The racemic compound tert-butyl - ((3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo ^] azulen-2-yl) methyl) carbamate (example 1 (g)) was separated supercritical fluid chromatography (SFC) (CHIRALCEL OJ-H chiral column, elution with MeOH (15%) and CO ₂ (85%)), and two chiral compounds (S) -tert-butyl - ((3-bromo-7 , 8-dihydro-2H-1,6,9-trioxa-9aborabenzo [cd] azulen-2-yl) methyl) carbamate (isomer eluting second, RT = 3.8 min) and (R) -isomer of tert-butyl - ((3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate (isomer eluting first, RT = 3.3 min ). Each of these chiral compounds (1.2 g, 3.13 mmol) in a saturated solution of HCl (gas) in Et ₂ O (20 ml) was stirred at room temperature for 1 h and concentrated to dryness (water bath <30 ° C) ... The residue was washed with acetonitrile (2x5 ml) and the white solid was dried under high vacuum to give the product (900 mg, 90%) as a white solid. MS (ERI) m / z = 284/286 [M + H] +.

(S) - (3-Bromo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride. 'and NMR (300 MHz, DMSO-d ₆ ) δ 8.40 (s, 3H), 7.63-7.61 (d, 1H), 6.92-6.89 (d, 1H), 5.50-5.48 (d, 1H), 4.68 (m , 1H), 4.35-4.22 (m, 3H), 3.60 (m, 1H), 3.00 (m, 1H).

(R) - (3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride. 1H NMR (300 MHz, DMSO-d ₆ ) δ 8.30 (s, 3H), 7.64-7.61 (d, 1H), 6.93-6.90 (d, 1H), 5.51-5.49 (d, 1H), 4.68 (m, 1H), 4.36-4.23 (m, 3H), 3.61 (m, 1H), 3.05-3.01 (m, 1H).

Method B.

(a) (S) -tert-Butyl - ((3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate

• —NHBoc Br -—NHBoc

A mixture of (S) -tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (110.0 g, 360, 50 mmol) (example 4, method B (h)) and NBS (67.4 g, 378.53 mmol) in DCE (1.1 L) were heated at 50 ° C for 6 h. The solution was washed with hot water (1 k) three times and the organic solution was concentrated in vacuo to give the title product (132.0 g, crude) as a yellow gum (used in the next step without purification). 1H NMR (400 MHz, DMSO-d ₆ ) 7.57-7.55 (d, J = 8 Hz, 1H), 6.96 (s, 1H), 6.85-6.83 (d, J = 8 Hz, 1H), 5.25 (m, 1H), 4.71-4.69 (m, 1H), 4.34-4.07 (m, 3H), 3.76-3.69 (m, 1H), 3.17-3.16 (m, 1H), 1.33 (s, 9H). LC / MS: [M-55] = 327.8.

(b) Header Specified Compound

A solution of (S) -tert-butyl - ((3-bromo-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate (130.0 d, unrefined) and conc. HCl (100 ml) in 1,4-dioxane (500 ml) was stirred at rt for 8 h, the precipitated colorless solid was filtered and washed with 2 propanol (200 ml). The solid was dried in vacuo at 50 ° C for 6 hours to give the hydrochloride salt of the desired product (60.0 g, 51.9% total yield over two steps) as a colorless solid. 1H NMR (400 MHz, DMSO-d ₆ ) 8.45 (s, 3H), 7.64-7.62 (d, J = 8, 1H), 6.92-6.90 (d, J = 8, 1H), 5.52 (m, 1H) , 4.69 (m, 1H), 4.37-4.15 (m, 3H), 3.74-3.50 (m, 1H), 3.05-2.95 (m, 1H). ¹³ C NMR (400 MHz, DMSO-d ₆ ) 161.80, 151.28, 137.57, 118.64, 107.18, 80.04, 73.86, 69.18, 41.88 LC / MS: [M + 1] ⁺ = 283.9.

Example 3. 3-Chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (G3-C1)

(a) 3-Chloro-2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene

In a solution of 2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene (29 g, 123.4

- 14 038093 mmol) (example 1 (d)) in DMF (250 ml) at 80 ° C was added a solution of NCS (16.5 g, 123.4 mmol) in DMF (100 ml). The mixture was stirred for 30 min at 80 ° C. The reaction mixture was poured into ice-water and extracted with EtOAc (200 ml × 3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by recrystallization from petroleum ether / ethyl acetate (10 1) to give 24 g of the crude product MS (ERI) m / z = 270 [M + H] +. ¹ H NMR (300 MHz, DMSO-d6) δ 7.52-7.49 (d, 1H), 6.99-6.96 (d, 1H), 5.96-5.93 (m, 1H), 5.42-5.30 (m, 1H), 4.80- 4.61 (m, 2H), 4.43-4.17 (m, 3H).

(b) Header Specified Compound

A solution of 3-chloro-2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene (24 g, 89.22 mmol), Raney Ni (4.0 d) and 7M NH ₃ in MeOH (20 ml) in methanol (300 ml) were shaken in an atmosphere of H ₂ for 2 h at rt. The mixture was filtered through a pad of Celite and the filtrate was concentrated in vacuo. The crude amine was dissolved in MeOH (20 ml) and concentrated HCl (5 ml) was added. The resulting mixture was stirred at rt for 1 h and then concentrated under reduced pressure. The resulting solid was washed with acetonitrile / hexanes (2 1, 2x200 ml) to give the title product (12 g, 50%) as a white solid. 1H NMR (300 MHz, DMSO-d ₆ ) δ 8.19 (s, 3H), 7.517.48 (d, 1H), 6.99-6.96 (d, 1H), 5.56-5.54 (d, 1H), 4.69 (m, 1H), 4.36-4.23 (m, 3H), 3.58 (m, 1H), 3.05-3.01 (m, 1H). MS (ERI) m / z = 240 [M + H] ⁺ .

Example 4-I. (S) - (3-Chloro-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (G4-Cl)

Method A.

(a) tert-Butyl - ((3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

In a mixture of (3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (8.0 g, 33.7 mmol) (example 3 (b)) and triethylamine (10.2 g, 101.2 mmol) in dichloromethane (250 ml) at 0 ° C., di-tert-butyl dicarbonate (11 g, 50.6 mmol) was added and the mixture was stirred for 2 h at kt. The reaction was quenched with saturated NaHCO ₃ solution (150 ml) and the resulting mixture was extracted with EtOAc (2x200 ml). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient to give the desired product (4.6 g, 47%). MS (ERI) m / z = 284 [M-56] ⁺ .

(b) Header Specified Compound

The racemic compound tert-butyl - ((3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate was separated by SFC (CHIRALCEL chiral column OJ-H), eluted with EtOH (15%) and CO ₂ (85%)) to give two chiral compounds (S) -tert-butyl - ((3-chloro-7,8dihydro-2H-1,6,9- trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (isomer eluting second, RT = 2.9 min) and (R) -tert-butyl ((3-chloro-7,8-dihydro- 2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2yl) methyl) carbamate (first eluting isomer, RT = 2.6 min). Each of these chiral compounds (4.6 g, 13.6 mmol) was stirred at rt in 80 ml of a saturated solution of HCl (gas) in Et ₂ O for 1 h and concentrated to dryness (water bath temperature <30 ° C). The residue was triturated with acetonitrile (2x5 ml) and the white solid was dried under high vacuum to give two products, (S) - (3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (1.2 g) and (R) - (3chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (2.3 g) respectively as white solids. MS (ERI) m / z = 240 [M + H] ⁺ .

(S) - (3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride. 1H NMR (300 MHz, DMSO-d6) δ 8.30 (s, 3H), 7.51-7.48 (d, 1H), 6.99-6.96 (d, 1H), 5.59-5.57 (d, 1H), 4.68 (m, 1H ), 4.36-4.23 (m, 3H), 3.58 (s, 1H), 3.03-2.99 (m, 1H).

- 15 038093

G4-CL- (R) (R) - (3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride. 1H NMR (300 MHz, DMSO-dg) δ 8.28 (s, 3H), 7.51-7.48 (d, 1H), 6.99-6.96 (d, 1H), 5.58-5.56 (d, 1H), 4.69 (m, 1H ), 4.36-4.23 (m, 3H), 3.59 (m, 1H), 3.05-3.01 (m, 1H).

Method B.

(a) (Z) -1- (Pyridin-2-yl) -N - ((1R) -1,7,7-trimethylbicyclo [2.2.1] heptan-2-ylidene) methanamine

A mixture of (+) - camphor (371 g, 2.44 mol), pyridin-2-ylmethanamine (277 g, 2.56 mol) and BF ₃ Et ₂ O (17 g, 0.12 mol) in toluene (3, 7 L) was loaded into a 5 L round bottom flask equipped with a DeanStark trap, reflux condenser, thermometer and nitrogen inlet. The mixture was heated to reflux with azeotropic removal of water for 20 h. The mixture was cooled to 15 ° С and quenched with 5% aqueous sodium bicarbonate solution (2.5 L), the organic phase was separated and washed with water (1.25 Lx2) then the mixture was concentrated to 2 L in vacuo. The residue was used in the next step without purification. 1H NMR (400 MHz, DMSO-d6) 8.47-8.48 (d, J = 4.4 Hz, 1H), 8.77-8.74 (t, J = 7.6 Hz, 1H), 7.43-7.41 (d, J = 8.0 Hz, 1H ), 7.25-7.22 (dd, J = 4.8 Hz, 1H), 4.49-4.38 (dd, J = 16.4 Hz, 2H), 2.46-2.42 (m, 1H), 1.97-1.93 (m, 2H), 1.841. 79 (m, 1H), 1.71-1.64 (m, 1H), 1.33-1.22 (m, 2H), 0.93 (s, 3H), 0.92 (s, 3H), 0.73 (s, 3H) LC / MS: [ M + H] ⁺ = 243.

(b) (1R) -1,7,7-trimethyl-N- (pyridin-2-ylmethyl) bicyclo [2.2.1] heptane-2-amine / - ^ 7 РУС / Н ₂ / toluene _r

Z --- An ^ /> -------- ► Λ-An />

5% Pt / C (40 g) was charged into a 5 liter pressure vessel, then the solution (Z) -1 - (pyridin-2-yl) -N - ((1R) -1,7,7- trimethylbicyclo [2.2.1] heptan-2-ylidene) methanamine (2.44 mol) in toluene (2 L). The vessel was pressurized with hydrogen at 100 psi (690 kPa) over a period of 12 hours. The solid was filtered through Celite® and the filter cake washed with toluene (1 L). The filtrate was concentrated in vacuo to give the desired product (435 g, 73% overall yield over two steps) as a pale yellow oil. 1H NMR (400 MHz, DMSO-d ₆ ) 8.498.48 (d, J = 4.8 Hz, 1H), 7.75-7.71 (t, J = 7.6 Hz, 1H), 7.40-7.38 (d, J = 7 6 Hz , 1H), 7.24-7.21 (dd, J = 5.2 Hz, 1H), 3.79-3.64 (dd, J = 14.4 Hz, 2H), 2.53-2.49 (m, 1H), 1.99 (s, 1H), 1.68- 1.42 (m, 5H), 1.05 (s, 3H), 0.87 (s, 3H), 0.78 (s, 3H). LC / MS: [M + H] ⁺ = 245.

(c) 3 - (2- (Benzyloxy) ethoxy) benzaldehyde

To a solution of 3-hydroxybenzaldehyde (2.90 kg, 23.75 mol) and ((2-bromoethoxy) methyl) benzene (4.26 kg, 19.79 mol) in DMF (9.3 L) was added K ₂ CO ₃ (3.83 kg, 27.70 mol). The reaction mixture was stirred at rt for 24 h. Water (15 L) and tert-butyl methyl ether (23 L) were added to the reaction mixture. The organic phase was separated and washed sequentially with 1N NaOH (2x 15 L) and water (15 L) and then concentrated to a minimum. Ethanol (23 L) was added and the solution concentrated in vacuo to give the title product (4.7 kg, 93%) as a colorless oil. 1H NMR (400 MHz, DMSO-d ₆ ) 9.98 (s, 1H), 7.55-7.52 (m, 2H), 7.46 (s, 1H), 7.36-7.34 (m, 4H), 7.32-7.26 (m, 2H ), 4.57 (s, 2H), 4.25-4.22 (t, J = 4.4 Hz, 2H), 3.80-3.78 (t, J = 4.4 Hz, 2H). LC / MS: [M + Na] ⁺ = 279.

(d) (S) -1 - (3 - (2- (benzyloxy) ethoxy) phenyl) -2-nitroethanol

A mixture of copper (II) acetate (167 g, 0.92 mol), (1R) -1,7,7-trimethyl-N- (pyridin-2ylmethyl) bicyclo [2.2.1] heptane-2-amine (269 g, 1.10 mol) in ethanol (19 L) was stirred at rt for 1 h, then a solution of 3- (2- (benzyloxy) ethoxy) benzaldehyde (4.70 kg, 18.34 mol) in ethanol (5 L) was added ... The reaction mixture was cooled to a temperature in the range of -30 to -40 ° C and then nitromethane (9.9 L, 183.40 mol) was added dropwise keeping the temperature below -30 ° C, after which diisopropylethylamine (285 g, 2 , 20 mol). The reaction mixture was stirred at -30 ° C for 24 h and then quenched with trifluoroacetic acid (314 g, 2.75 mol). To the resulting solution were added 1N HCl (24 L) and TBME (47 L). The separated organic phase was washed with water (24 L) and then concentrated in vacuo. The residue was added to petroleum ether / ethyl acetate = 5: 1 (10 L). Then the precipitated yellow solid was collected by filtration using a Buchner funnel and dried in vacuo at 40 ° C for 6 h to give the title product (5.00 kg, 86%) as a white solid.

- 16 038093 substances. 1H NMR (400 MHz, DMSO-d6) 7.38-7.25 (m, 6H), 7.03 (s, 1H), 7.01-6.99 (d, J = 7.6 Hz, 1H), 6.90

6.87 (dd, J = 8.0 Hz, 1H), 6.09-6.08 (d, J = 5.2 Hz, 1H), 5.26-5.22 (m, 1H), 4.86-4.82 (dd, J = 12.4 Hz, 1H), 4.574 .51 (m, 3H), 4.15-4.13 (m, 2H), 3.78-3.76 (t, J = 4.8 Hz, 2H). LC / MS: [M + Na] + = 340.

(f) (S) -1- (3- (2- (benzyloxy) ethoxy) phenyl) -2- (dibenzylamino) ethanol hydrochloride

10% Pd / C (800 g) and 10% Pt / C (200 g) were charged into a pressure vessel, then a solution of (S) -1- (3- (2- (benzyloxy) ethoxy) phenyl) - 2-nitroethanol (5.00 kg, 15.76 mol) in ethanol (50 L). The vessel was pressurized with 100 psi (690 kPa) hydrogen over a period of 12 hours at rt. The solid was filtered through Celite® and the filter cake washed with ethanol (5 L). _{K 2} CO ₃ (4.80 kg, 34.67 mol) and benzyl bromide (5.93 kg, 34.67 mol) were sequentially added to the filtrate. The reaction mixture was stirred at rt for 24 h. The solid was filtered off and washed with ethanol (1 L). The filtrate was diluted with water (20 L) and then heated to 50 ° C. The solution was stirred at 50 ° C for 30 min and then conc. HCl (1.5 L) for 1 h. The mixture was cooled to 0 ° C and held at 0 ° C for an additional 30 min. The product was filtered and washed with 20% aqueous ethanol solution (1 L) to give the hydrochloride salt of the title product (5.00 kg, 63% over two steps) as a colorless solid. 1H NMR (400 MHz, DMSO-d ₆ ) 10.67 (s, 1H), 7.72-7.68 (m, 4H), 7.47-7.45 (m, 6H), 7.38-7.26 (m, 5H), 7.25-7.21 (t , J = 7.6 Hz, 1H), 6.86-6.84 (d, J = 8.0 Hz, 1H), 6.77 (s, 1H), 6.77-6.75 (d, J = 7.2 Hz, 1H), 6.36 (s, 1H) , 5.04-5.02 (d, J = 9.2 Hz, 1H), 4.58 (s, 2H), 4.51-4.38 (m, 4H), 4.09-4.07 (t, J = 4.0 Hz, 2H), 3.77-3.75 (t , J = 3 2 Hz, 2H), 3.13-2.96 (m, 2H). LC / MS: [M + H] + = 468.

(f) (S) -7- (2- (benzyloxy) ethoxy) -3 - ((dibenzylamino) methyl) benzo [c] [1,2] oxaborol-1 (3H) -ol

To a solution of (S) -1- (3- (2- (benzyloxy) ethoxy) phenyl) -2- (dibenzylamino) ethanol hydrochloride (3.85 kg, 7.64 mol) in dry toluene (39 L) at -30 ° C under N2, n-BuLi (15.3 L, 38.20 mol) was added dropwise over 6 h. After the addition, the mixture was stirred at -30 ° C for an additional 1 h and then cooled to -70 ° C. Trimethyl borate (3.97 kg, 38.20 mol) was added dropwise keeping the temperature below -60 ° C. After the addition, the reaction mixture was left to warm to rt and was stirred overnight. The reaction mixture was quenched with 5% aqueous NaHCO ₃ solution (20 L) and stirred vigorously for 15 min. The resulting suspension was filtered and the filtrate was separated. The organic layer was washed with water (20 L x 3) and concentrated in vacuo and the residue was purified by chromatography on silica gel eluting with 5: 1 petroleum ether / ethyl acetate to give the title product (1.80 kg, 48%) as a yellow solid. 1H NMR (400 MHz, DMSO-d ₆ ) 8.81 (s, 1H), 7.39-7.22 (m, 16H), 6.82-6.80 (d, J = 7.6 Hz, 1H), 6.72-6.70 (d, J = 7.6 Hz, 1H), 5.34-5.31 (dd, J = 7.6 Hz, 1H), 4.60 (s, 2H), 4.22-4.19 (t, J = 4.4 Hz, 2H), 3.80-3.72 (m, 6H), 2.88 -2.84 (dd, J = 13.6 Hz, 1H), 2.47-2.45 (dd, J = 10 Hz, 1H). LC / MS: [M + H] + = 494.

(g) (S) - (7,8-Dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride

10% Pd / C (180 g) was charged into a pressure vessel, then a solution of (S) -7- (2 (benzyloxy) ethoxy) -3 - ((dibenzylamino) methyl) -benzo [c] [1, 2] -oxaborol-1 (3H) -ol (1.80 kg, 3.65 mol) in methanol (18 L), toluene (3.6 L) and 1 g HCl (4 L). The vessel was pressurized with hydrogen at a pressure of 100 psi. inch over a period of 12 hours at 50 ° C. The solid was filtered through Celite and the filter cake was washed with methanol (1 L). The filtrate was concentrated in vacuo, the residue was treated with 2-propanol (3.6 L) and stirred at rt for 30 min. The resulting solid was collected by filtration, washed with 2-propanol (500 ml) and dried in vacuo at 50 ° C for 6 h to give the title product (680 g, 77%) as a pale yellow powder. ¹ H NMR (400 MHz, DMSO-d ₆ ) 8.38 (s, 3H), 7.52-7.48 (t, J = 8.0 Hz, 1H), 7.17-7.15 (d, J = 7.6 Hz, 1H), 6.92-6.90 (d, J = 7.6 Hz, 1H), 5.55 (m, 1H), 4.71-4.68 (m, 1H), 4.38-4.22 (m, 3H), 3.53-3.50 (m, 1H), 2.91-2.86 (m , 1H). LC / MS: [M + H] + = 206.

(h) (S) -tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

- 17 038093

In a solution of (S) - (7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (390 g, 1.62 mol) and Et ₃ N (163.4 g, 4.85 mol) in DCM (4 6 L) was added dropwise (Boc) ₂ O (353.0 g, 1.62 mol) over 2 h at rt. After the addition, the reaction mixture was stirred at rt for an additional 3 h. The reaction was quenched with 1 N HCl (4 L) and the organic phase was separated, washed with water (4 L) and concentrated in vacuo to give the desired product (460 g, 93%) as a pale white solid. ¹ H NMR (400 MHz, DMSO-d6) 7.46-7.42 (t, J = 7.6 Hz, 1H), 7.07 (s, 1H), 7.02-7.00 (d, J = 7.2 Hz, 1H), 6.876.85 ( d, J = 8.0 Hz, 1H), 5 27 (m, 1H), 4.68-4.65 (m, 1H), 4.34-4.18 (m, 3H), 3.41 (s, 1H), 3.14-3.08 (m, 1H ), 1.38 (s, 9H). LC / MS: [M-55] = 250.

(i) (S) -tert-butyl - ((3-chloro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

A mixture of (S) -tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate (315.0 g, 1, 03 mol) and NCS (144.5 g, 1.08 mol) in dichloroethane (3.5 L) were heated at 50 ° C for 24 h.

The solution was washed with hot water (50 ° C, 4 L x 3) and the organic phase was concentrated in vacuo to give the title product (400.0 g, crude) as a yellow solid, which was used in the next step without further purification. ¹ H NMR (400 MHz, DMSO-d ₆ ) 7.44-7.42 (d, J = 8.4 Hz, 1H), 6.99 (s, 1H), 6.91-6.89 (d, J = 8.4 Hz, 1H), 5.33 (m , 1H), 4.72-4.69 (m, 1H), 4.35-4.19 (m, 3H), 3.73-3.71 (m, 1H), 3.17-3.15 (m, 1H), 1.33 (s, 9H). LC / MS: [M-55] = 284.

(j) Header Specified Compound

A solution of (S) -tert-butyl - ((3-chloro-7,8-dihydro-2Н-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate (400, 0 g, unrefined) and conc. HCl (500 ml) in 1,4-dioxane (2 L) was stirred at rt for 8 h, the precipitated colorless solid was collected and washed with 2 propanol (200 ml). The solid was recrystallized from H ₂ O and dioxane (400 ml / 2000 ml) to give the hydrochloride salt of the title product (110.0 g, 39% over two steps). 1H NMR (400 MHz, DMSO-d ₆ ) 8.48-8.35 (br, 3H), 7.52-7.50 (d, J = 8.8 Hz, 1H), 7.00-6.97 (d, J = 8.4 Hz, 1H), 5.60 ( m, 1H), 4.71 (m, 1H), 4.38-4.21 (m, 3H), 3.64-3.55 (m, 1H), 3.04-2.99 (m, 1H). ¹³ C NMR (400 MHz, DMSO-d6) 161.22, 149.15, 134.61, 119.35, 118.31, 79.14, 73.92, 69.22, 41.88. LC / MS: [M + H] + = 240.

Example 4-II. (S) - (3-Chloro-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methanamine dihydrosulfate · H ₂ O (G4-CL)

T b H ₂ SO ₄ H ₂ O ^Cl nh ₂

A mixture of (S) -tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (13.25 kg) and NCS (8.75 kg) in dichloroethane (132.5 L) was heated at 70 ° C until the reaction was complete by HPLC. The mixture was concentrated under reduced pressure, cooled to 25 ° C and acetone (106 L) added. The suspension was filtered, rinsing with acetone (26.5 L). The wet filter cake was suspended in water (13.25 L) and 1,4-dioxane (66.25 L), heated to 50 ° C for 20-30 min, cooled to 15 ° C, filtered and the filter cake was washed 1,4-dioxane (26.5 L). The wet filter cake was dissolved in methanol (68.9 L), filtered, and the filtrate was concentrated under reduced pressure. Methyl tert-butyl ether (66.25 L) was added to the residue, and the mixture was concentrated under reduced pressure. Methyl tert-butyl ether (78.7 L), isopropanol (8.7 L) and sulfuric acid (4.6 L) were added, the mixture was heated to 50 ° C and stirred until the sulfate content was 24.32-29.72% ... The mixture was cooled to 25 ° C, stirred for 1 h, filtered and the filter cake was washed with methyl tert-butyl ether (17.5 L) and dried to give the title product (42%).

Example 5 (comparative). (3-Fluoro-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2yl) methanamine hydrochloride) (G5-F)

- 18 038093 (a) 1 - (2- (benzyloxy) ethoxy) -2-bromo-4-fluorobenzene

A solution of 2-bromo-4-fluorophenol (1.91 g, 10 mmol), ((2-bromoethoxy) methyl) benzene (2.6 g, 12 mmol) and

K ₂ CO ₃ (2.76 g, 20 mmol) in 40 ml DMF was stirred at 25 ° C for 16 h. Then the mixture was poured into 300 ml of water, extracted with ethyl acetate (200 ml), washed with water (200 ml) and brine (100 ml) and dried over anhydrous sodium sulfate. The solvent was evaporated at 40 ° C. under reduced pressure and the residue was purified by silica gel chromatography eluting with ethyl acetate and petroleum ether (1: 5) to give the product (3.1 g, 95%) as a colorless oil. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.55 (dd, 1H), 7.36-7.15 (m, 7H), 4.60 (s, 2H), 4.22-4.19 (m, 2H), 3.80-3.77 (m , 2H).

(b) 3- (2- (Benzyloxy) ethoxy) -2-bromo-6-fluorobenzαldehyde

A solution of 1- (2- (benzyloxy) ethoxy) -2-bromo-4-fluorobenzene (1.6 g, 4.9 mmol) in 30 ml THF was cooled to -70 ° C and LDA (2.0 M in THF, 3.5 ml, 7 mmol). The resulting mixture was stirred for 2 h at low temperature, after which a solution of DMF (1.1 g, 15 mmol) in THF (3 ml) was added. The mixture was stirred for 1 hour and then allowed to warm to 0 ° C. The mixture was quenched with saturated aqueous NH4Cl and the mixture was extracted with ethyl acetate (100 ml). The organic layer was washed with water (50 ml) and brine (50 ml) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and the residue was purified by silica gel chromatography eluting with ethyl acetate and petroleum ether (1-3) to give the product (1.2 g, 69%) as a colorless oil. 1H NMR (300 MHz,

DMSO-d6) δ 10.22 (s, 1H), 7.48-7.27 (m, 7H), 4.60 (s, 2H), 4.29-4.26 (m, 2H), 3.82-3.79 (m, 2H).

(c) 6- (2- (Benzyloxy) ethoxy) -3-fluoro-2-formylphenylboronic acid

Solution of 3- (2- (benzyloxy) ethoxy) -2-bromo-6-fluorobenzaldehyde (1 g, 2.8 mmol), Pin ₂ B ₂ (1 g, 4 mmol), KOAc (0.56 g, 6 mmol ) and Pd (dppf) Cl ₂ (0.05 g) in 30 ml THF were degassed using N ₂ six times. The mixture was then heated at 100 ° C (under microwave irradiation) for 4 hours. The reaction mixture was cooled to room temperature, filtered and concentrated under reduced pressure. The residue was purified by chromatography on silica gel, eluting with ethyl acetate and petroleum ether (1: 5). The fractions were combined and concentrated under reduced pressure. The residue was dissolved in THF (20 ml) and 6N HCl (4 ml) and the resulting mixture was stirred at room temperature for 1 hour. After extraction with ethyl acetate (20 ml × 3), the combined organic layer was concentrated under reduced pressure to give the crude product (0.5 d, 56%). It was used directly in the next step without further purification. LC / MS: 336.0 [M + H ₂ O] +.

(d) 7- (2- (Benzyloxy) ethoxy) -4-fluoro-3- (nitromethyl) benzo [c] [1,2] oxaborol-1 (3H) -ol

To a stirred solution of 6- (2- (benzyloxy) ethoxy) -3-fluoro-2-formylphenylboronic acid (0.5 g, 1.6 mmol) and CH3NO2 (0.2 g, 3.5 mmol) in 10 ml THF a solution of NaOH (0.028 g, 0.7 mmol) in 3 ml of water was added at room temperature. Then the mixture was stirred at room temperature for 16 h and acidified with conc. HCl to pH 1 at 0 ° C. The mixture was extracted with ethyl acetate (20 ml) and the organic layer was washed with water (10 ml) and brine (10 ml), then dried over anhydrous sodium sulfate. After removal of the solvent under reduced pressure, the residue was purified by silica gel chromatography eluting with ethyl acetate and petroleum ether (1:10) to give the crude product (0.5 g, 88%) as a colorless oil. LC / MS: 379.0 [M + H ₂ O] ⁺ .

(f) Header Compound

- 19 038093

A solution of 7- (2- (benzyloxy) ethoxy) -4-fluoro-3 - (nitromethyl) benzo [c] [1,2] -oxaborol-1 (3H) -ol (0.5 g, 1.4 mmol) and Pd / C (10%, 0.1 g) in 20 ml of methanol was hydrogenated under 1 atm of H ₂ at room temperature for 48 hours. It was then filtered through a Celite pad and the filtrate was concentrated under reduced pressure to give an oil. The crude product was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient. The collected fraction was concentrated under reduced pressure. The residue was dissolved in diethyl ether (5 ml) and 2N HCl (0.2 ml) was added. The resulting mixture was stirred at room temperature for 1 hour. The solid was collected by filtration and washed with diethyl ether (10 ml) to give the title compound (0.035 g, 10%) as a white solid. LC / MS: 223.9 [M + H] +. L NMR (400 MHz, DMSO-d ₆ ) δ 8.22 (brs, 3H), 7.33 (t, 1H), 6.97 (dd, 1H), 5.68 (d, 1H), 4.69 (brs, 1H), 4.37-4.23 (m, 3H), 3.43-3.40 (m, 1H), 3.03 (t, 1H).

Example 6 (comparative). (S) - (3-Iodo-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methanamine hydrochloride (G6-I)

CX> HCI

I 'NH ₂ (a) (S) -tert-butyl - ((3-iodo-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl ) carbamate θ ^ζ о р ⁷ о

A. in NIS, SPLAS K.T., 24 hours A. _in '

Co ---- Qo '-NHBoc I' -NHBoc

A solution of (S) -tert-butyl - ((7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (300 mg, 0.98 mmol ) (example 4, method B (h)) and NIS (265 mg, 1.18 mmol) in 6 ml of AcOH were stirred at room temperature for 24 hours. The solvent was evaporated at 40 ° C. under reduced pressure. The residue was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient to give the product (200 mg, 47%) as a light yellow oil. LC / MS: 432 [M + H] +.

(b) Header Specified Compound

A solution of (S) -tert-butyl - ((3-iodo-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (140 mg , 0.32 mmol) and TFA (0.5 ml) in 5 ml of DCM was stirred at room temperature for 2 hours. The solvent was evaporated at 40 ° C under reduced pressure. The residue was dissolved in diethyl ether (5 ml) and 2N HCl in water (0.2 ml) was added. The resulting mixture was stirred at room temperature for 15 minutes. The solid was collected by filtration and washed with diethyl ether (10 ml) to give the title compound (90 mg, 75%) as a white solid. LC / MS: 332.0 [M + H] ⁺ . L NMR (400 MHz, DMSO-d ₆ ) δ 8.47 (brs, 3H), 7.80 (d, 1H), 6.78 (d, 1H), 5.37 (m, 1H), 4.72-4.53 (m, 1H), 4.49 -4.08 (m, 3H), 3.78-3.51 (m, 1H), 3.06-2.78 (m, 1H).

Example 7 (comparative). (3-Chloro-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen2-yl) methanamine hydrochloride (G7-CI)

(a) 2-Bromo-3 - (2-hydroxypropoxy) benzaldehyde

A solution of 2-bromo-3-hydroxybenzaldehyde (6.0 g, 29.85 mmol), 1-chloropropan-2-ol (8.46 g, 89.55 mmol) and K ₂ CO ₃ (8.24, 59, 7 mmol) in DMF (100 ml) was stirred at 100 ° C overnight. The reaction mixture was then quenched by the addition of water (4 L) and then extracted with EtOAc (3x1.5 L). The combined organic layers were washed with brine (250 ml), dried over anhydrous Na ₂ SO ₄ and concentrated to dryness in vacuo. The residue was purified by silica gel column chromatography (petroleum ether ethyl acetate = 5: 1 to 2: 1) to obtain the target crude compound (8.77 g). MS (ERI) m / z = 259/261 [M + H] ⁺ .

(b) 3- (2-Hydroxypropoxy) -2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) benzaldehyde

- 20 038093 .on. On o-Λ o Pin ₂ B ₂ , KOAc T '

------ st ° '

Pd (dppf) CI ₂ dioxane Π _{/ 0}

A solution of 2-bromo-3- (2-hydroxypropoxy) benzaldehyde (8.77 g, 34 mmol) 4,4,4 ', 4', 5,5,5 ', 5'-octamethyl 2,2'-bi- ( 1,3,2-dioxaborolane) (17.27 g, 68 mmol), Pd (dppf) Cl ₂ (2.49 g, 3.4 mmol) and KOAc (9.99 g, 102 mmol) in dioxane (200 ml) was stirred at 100 ° C overnight. The reaction mixture was then quenched by the addition of water (200 ml) and then extracted with EtOAc (3x200 ml). The combined organic layers were washed with brine (250 ml), dried over anhydrous Na ₂ SO ₄ and concentrated to dryness in vacuo. The residue was purified by silica gel column chromatography (petroleum ether ethyl acetate = 5: 1 to 1: 1) to obtain the target crude compound (6 g). MS (ERI) m / z = 307 [M + H] ⁺ .

(c) 8-Methyl-2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene

H ₃ NO ₂ , NaOH thf / h ₂ o no ₂

To a solution of NaOH (261.4 mg, 6.54 mmol) in water (8 ml) was added nitromethane (1.2 g, 19.6 mmol) at 5-10 ° C. After stirring for 15 min at 5-10 ° C, CTAB (0.19 g, 0.52 mmol) was added to the reaction mixture and then 3- (2-hydroxypropoxy) -2- (4,4,5,5- tetramethyl-1,3,2-dioxaborolane-2yl) benzaldehyde (2.0 g, 6.54 mmol) at 5-10 ° C. The reaction mixture was stirred at rt for 5 h. The reaction mixture was acidified to pH 1 using dilute hydrochloric acid and stirred at rt overnight. The reaction mixture was filtered to give the title compound (541 mg, 33%).

(d) (8-Methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine acetate

A solution of 8-methyl-2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene (541 mg, 2.173 mmol) and palladium hydroxide (300 mg) in acetic acid (10 ml) was shaken under an atmosphere of H ₂ overnight at room temperature. The mixture was filtered through a Celite pad and the filtrate was concentrated in vacuo to give the crude compound (350 mg). MS (ERI) m / z = 220 [M + H] ⁺ .

(f) tert-Butyl - ((8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

Into a mixture of the crude compound (8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen2-yl) methanamine acetate (3.0 g, 10.75 mmol) and triethylamine (6.5 g, 64.5 mmol) in dichloromethane (100 ml) at 0 ° C was added di-tert-butyl dicarbonate (3.5 g, 16.13 mmol) and the mixture was stirred for 2 h at room temperature. The reaction was quenched with saturated NaHCO ₃ (15 ml) and the resulting mixture was extracted with EtOAc (3x80 ml). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient to give the product (700 mg). MS (ERI) m / z = 264 [M-56] ⁺ . Ή NMR (300 MHz, DMSO-d ₆ ) δ 7.44-7.39 (m, 1H), 7.01-6.98 (m, 2H), 6.886.85 (m, 1H), 5.24 (m, 1H), 4.52-4.44 ( m, 2H), 4.18-4.00 (m, 1H), 3.39-3.36 (m, 1H), 3.15-3 06 (m, 1H), 1.421.09 (m, 15H).

(f) tert-Butyl - ((3-chloro-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

solution of tert ^, -butyl - ((8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2 yl) methyl) carbamate (300 mg, 0.94 mmol ) and 1-chloropyrrolidine-2,5-dione (151.4 mg, 1.13 mmol) in CH ₃ CN (20 ml), 2,2'-azobis- (2-methylpropionitrile) (15.4 mg, 0.094 mmol) and the mixture was stirred for 2 h at 90 ° C. The reaction mixture was then concentrated under high vacuum and the residue was purified by preparative HPLC using a Gemini® 5 μ C18 column (150x21.2 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient to afford the desired product (150 mg, 45 %). MS (ERI) m / z = 298 [M-56] ⁺ .

- 21 038093 (g) Title Compound

tert-Butyl - ((3-chloro-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo ^] azulen-2-yl) methyl) carbamate (150 mg, 0.425 mmol ) in Et ₂ O / HCl and Et ₂ O (10 ml) was stirred at room temperature for 2 h and concentrated to dryness (water bath <30 ° C). The residue was washed with acetonitrile (2x5 ml) and the white solid was dried under high vacuum to give the product (120 mg, 97%) as a white solid. ¹ H NMR (300 MHz, DMSO-d6) δ 8.28 (s, 3H), 7.51-7.48 (d, 1H), 7.02-6.99 (d, 1H), 5.58-5.56 (d, 1H), 4.57 (m, 2H), 4.33-4.17 (m, 1H), 3.72-3.56 (m, 1H), 3.05-3.01 (m, 1H), 1.30-1.23 (m, 3H). MS (ERI) m / z = 254 [M + H] +.

Example 8 (comparative). (3-Bromo-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen2-yl) methanamine hydrochloride (G8-Br)

HCI nh ₂ (a) tert-Butyl - ((3-bromo-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate

In a solution of tert-butyl - ((8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2yl) methyl) carbamate (180 mg, 0.564 mmol) (example 5 (f)) and 1-bromopyrrolidine-2,5-dione (120 mg, 0.677 mmol) in CH ₃ CN (20 ml), 2,2'-azobis- (2-methylpropionitrile (9.2 mg, 0.056 mmol ) and the mixture was stirred for 2 h at 90 ° C. The reaction mixture was then concentrated in high vacuum and the residue was purified by preparative HPLC using a Gemini® 5μ C18 column (150x21.2 mm) and eluting with a water / acetonitrile gradient (0.05% TFA) to give the product (60 mg) MS (ESI) m / z = 342/344 [M-56] ⁺ .

(b) Header Specified Compound

tert-Butyl - ((3-bromo-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate (60 mg, 0.15 mmol) in a saturated solution of HCl (gas) in Et ₂ O (10 ml) was stirred at rt for 2 h and concentrated to dryness (water bath temperature <30 ° C). The residue was purified by preparative HPLC using a Gemini® 5 μ C18 column (150x21.2 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient to give the product (20 mg) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 8.12 (br, 3H), 7.65 (m, 1H), 6.96 (m, 1H), 5.45 (m, 1H), 4.58 (m, 2H), 4.29-4.16 ( m, 1H), 3.77-3.59 (m, 1H), 3.04 (m, 1H), 1.29-1.21 (d, 3H). MS (ERI) m / z = 298/300 [M + H] ⁺ .

Example 9 (comparative). (3-Bromo-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (G9-Br)

(a) 2-Bromo-3- (2-hydroxy-2-methylpropoxy) benzaldehyde

A solution of 2-bromo-3-hydroxybenzaldehyde (7.5 g, 37.3 mmol), 1-chloro-2-methylpropan-2-ol (9.4 g, 85.6 mmol) and Na ₂ CO ₃ (6, 7 g, 63.2 mmol) in 70 ml of DMSO was stirred at 140 ° C for 3 h. Then the mixture was cooled to room temperature, poured into 300 ml of water, extracted with ethyl acetate (600 ml), washed with water (300 ml), brine (50 ml), dried over anhydrous sodium sulfate. The solvent was evaporated at 40 ° C under reduced pressure and the residue was purified by silica gel chromatography eluting with a mixture of ethyl acetate and petroleum ether (1: 3) to give the title compound

- 22 038093 nia (9.2 g, 90.3%) as a colorless oil. Ή NMR (300 MHz, CDCl ₃ ) δ 10.43 (s, 1H), 7.54 (dd, 1H, J1 = 3.0, J2 = 7.5), 7.40 ~ 7.34 (m, 1H), 7.54 (dd, 1H, J1 = 3 , J2 = 7.5), 3.90 (s, 2H), 1.42 (s, 6H).

(b) 3- (2-Hydroxy-2-methylpropoxy) -2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) benzaldehyde —Loh —Loh

Ϊ _Br Pin ₂ B ₂ , KOAc ΐΑθΚ

Pd (dppf) CI ₂ UY o o

A solution of 2-bromo-3- (2-hydroxy-2-methylpropoxy) benzaldehyde (9.2 g, 33.7 mmol), Pin ₂ B ₂ (17.1 g, 67.4 mmol), KOAc (9.9 g, 101.1 mmol) and Pd (dppf) Cl ₂ (2.5 g) in 240 ml of 1,4-dioxane were degassed with N2 six times. Then the reaction mixture was stirred at 99 ° C under nitrogen atmosphere for 16 h. The reaction mixture was cooled, filtered, then evaporated at 40 ° C under reduced pressure and the residue was purified by chromatography on silica gel, eluting with a mixture of ethyl acetate and petroleum ether (1: 5), to give the title compound (10 g, crude) containing the de-Br by-product (used directly in the next step without further purification).

(c) 8,8-Dimethyl-2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene

Into a stirred solution of 3- (2-hydroxy-2-methylpropoxy) -2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) benzaldehyde (10 g, 31.3 mmol) and CH3NO2 (5.7 g, 93.8 mmol) in 100 ml of THF was added a solution of NaOH (1.25 g, 31.3 mmol) in 60 ml of water at room temperature. Then the reaction mixture was stirred at room temperature for 16 hours. Then the reaction mixture was acidified with conc. HCl to pH 1 at 0 ° C and stirred at room temperature for 1 h. The mixture was extracted with ethyl acetate (100 ml), washed with water (30 ml), then brine (30 ml) and dried over anhydrous sodium sulfate. The solvent was evaporated at 40 ° C. under reduced pressure and the residue was purified by silica gel chromatography eluting with a mixture of ethyl acetate and petroleum ether (1:10) to give the title compound (3 g, 36.5%) as a colorless oil.

(d) (8,8-Dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine acetate o ^z

A. In Pd (OH) ₂ , HOAc AxA ί | Ί Ъ ----------- ► [| | About ^AsON

AA - / on AAa '-no ₂ ^ -mn ₂

A solution of 8,8-dimethyl-2- (nitromethyl) -7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene (1 g, 3.8 mmol) and Pd (OH) ₂ (10%, 0.2 g) in 20 mL acetic acid was hydrogenated at 1 atm H ₂ at rt for 16 h. The mixture was then filtered and the solvent was evaporated at 40 ° C under reduced pressure to give the title compound (0 , 9 g, crude) as an oil (acetate salt). LC / MS: 234.1 [M + H] +.

(f) tert-Butyl - ((8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methyl) carbamate

In a stirred solution of (8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2yl) methanamine acetate (0.7 g, 2.39 mmol) in 70 ml CH ₂ Cl ₂ cooled to 0 ° C was added Et ₃ N (0.61 g, 6.0 mmol). _{Then Boc 2} O (0.98 g, 4.5 mmol) was added in one portion and the reaction mixture was stirred at room temperature for 16 h. The mixture was washed with 0.3 N HCl (30 ml), water (30 ml) and dried over anhydrous sodium sulfate. The solvent was evaporated at 40 ° C. under reduced pressure and the residue was purified by silica gel chromatography eluting with a mixture of ethyl acetate and petroleum ether (14) to give the title compound (0.63 g, 79%) as an oil. LC / MS: 234.1 [M + H100] +.

(f) tert-Butyl - ((3-bromo-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2 yl) methyl) carbamate

A solution of tert-butyl - ((8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methyl) carbamate (232 g, 0.70 mmol), NBS (143 mg, 0.80 mmol) and AIBN (20 mg) in 30 ml of acetonitrile

- 23 038093 was stirred at reflux for 1 h. The solvent was evaporated at 40 ° C under reduced pressure and the residue was purified by chromatography on silica gel eluting with a mixture of ethyl acetate and petroleum ether (1 4) to give the title compound (260 mg, 88 , 6%) as a solid. LC / MS: 312.0 / 314.0 [M + H-100] +.

(g) Title compound

A solution of tert-butyl - ((3-bromo-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methyl) carbamate (260 mg, 0.63 mmol) in a saturated solution of HCl in 1,4-dioxane (20 ml) was stirred at room temperature for 3 hours.The solvent was evaporated at 40 ° C under reduced pressure and the residue was purified by preparative HPLC using a Gemini® 5 μ C18 column (150x21.2 mm), eluting with a water / acetonitrile gradient (0.05% TFA) and treating with 0.1 ml concentrated HCl to give the title product (20 mg, 9.1%) as a white solid. LC / MS: 311.9 [M + H] ⁺ . 1H NMR (400 MHz, CD3OD) δ 7.63 (d, 1H, J = 8), 6.95 (d, 1H, J = 8), 5.52-5.45 (m, 1H), 4.41 (d, 1H, J = 12) , 4.17 (d, 1H, J = 16), 4.09 ~ 3.85 (m, 1H), 3.13-2.98 (m, 1H), 1.37-1.30 (m, 6H).

Example 10 (comparative). (S) - (3-Bromo-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9aborabenzo [cd] azulen-2-yl) methanamine hydrochloride (G10-Br)

(a) (S) -tert-Butyl - ((3-bromo-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methyl ) carbamate

A solution of tert-butyl - ((8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methyl) carbamate (5.5 g, 16 , 5 mmol) (example 9 (f)) and NBS (3.2 g, 18.2 mmol) in 100 ml of dichloroethane was stirred at 50 ° C for 18 h. The solvent was evaporated at 40 ° C under reduced pressure and the residue was purified chromatography on silica gel eluting with a mixture of ethyl acetate and petroleum ether (1-10) to give the title compound (5.9 g, 86.5%) as an oil. The racemic compound was separated by SFC (chiral column CHIRALPAK AD-H, elution with EtOH (20%) and CO ₂ (80%)) to give 2.2 g of the (S) -isomer (isomer eluting first, RT = 3.0 min ) and 2.2 g of the (R) -isomer (isomer eluting second, RT = 4.1 min). LC / MS: 312.0 / 314.0 [M + H-100] ⁺ .

(b) Header Specified Compound

Dry HCl gas was bubbled through a solution of (S) -tert-butyl - ((3-bromo-8,8-dimethyl-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] azulene-2- yl) methyl) carbamate (2.2 g, 5.34 mmol) in diethyl ether (150 ml) at room temperature for 3 h and the mixture was then stirred for 18 h. The solution was filtered and the filter cake dried in vacuo to give (S) -isomer (1.4 g, 76%) as a white solid. LC / MS 311.9 [M + H] ⁺ . 1H NMR (400 MHz, DMSO-d6) δ 8.23 (brs, 3H), 7.64 (d, 1H, J = 8), 6.96 (d, 1H, J = 8), 5.48-5.46 (m, 1H), 4.43 -4.40 (m, 1H), 4.21 ~ 4.10 (m, 1H), 3.75 ~ 3.55 (m, 1H), 3.05 ~ 2.95 (m, 1H), 1.36 ~ 1.27 (ds, 6H). Similarly, acid treatment with (R) -tert-butyl - ((3-bromo-8,8dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2- yl) methyl) carbamate gave the corresponding (R) -isomer as a white solid (1.4 g, 76%). LC / MS: 312.0 [M + H] ⁺ . 1H NMR (400 MHz, DMSO-d6) δ 8.29 (brs, 3H), 7.65 (d, 1H, J = 8), 6.96 (d, 1H, J = 8), 5.48 ~ 5.46 (m, 1H), 4.42 -4.39 (m, 1H), 4.22 ~ 4.10 (m, 1H), 3.75 ~ 3.50 (m, 1H), 3.03 ~ 2.93 (m, 1H), 1.36 ~ 1.27 (ds, 6H).

Example 11 (comparative). (3-Chloro-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine hydrochloride (G11-CI)

(a) mret-Butyl - ((3-chloro-8,8-dimethyl-7,8-dihydro-2H-1,6,9-mrioxa-9а-borabenzo [cd] αzulene-2-24 038093 yl) methyl ) carbamate

A solution of tert-butyl - ((8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo ^] azulen-2-yl) methyl) carbamate (519 mg, 1.56 mmol) (example 9 (f)), NCS (250 mg, 1.87 mmol) and AIBN (30 mg) in 50 ml of acetonitrile was stirred at reflux temperature for 1 hour. The solvent was evaporated at 40 ° C under reduced pressure and the residue purified by chromatography on silica gel eluting with a mixture of ethyl acetate and petroleum ether (1: 5) to give the title product (300 mg, 52.4% containing the 6Cl isomer) as a solid. LC / MS: 268.1 [M + H-100] +.

(b) Header Specified Compound

A solution of tert-butyl - ((3-chloro-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo ^] azulene-2yl) methyl) carbamate (300 mg, 0 , 82 mmol) in a saturated solution of HCl in 1,4-dioxane (30 ml) was stirred at room temperature for 3 h.The solvent was evaporated at 40 ° C under reduced pressure, and the residue was purified by preparative HPLC using a Gemini® 5μ X18 column (150x21 , 2 mm), eluting with a gradient mixture of water / acetonitrile (0.05% TFA), then treated with 0.1 ml of conc. HCl to give the title product (94 mg, 37.9%) as a white solid. LC / MS: 268.1 [M + H] +. 1H NMR (400 MHz, DMSO-d ₆ ) δ 8.40 (brs, 3H), 7.52 (d, 1H, J = 8), 7.02 (d, 1H, J = 8), 5.60-5.58 (m, 1H), 4.42-4.38 (m, 1H), 4.23-4.07 (m, 1H), 3.67-3.57 (m, 1H), 3.02-2.92 (m, 1H), 1.36-1.27 (m, 6H).

Example 12 (comparative). (S) - (3-Chloro-8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] αsulen-2-yl) methanamine hydrochloride (G12-Cl)

ci -nh ₂ (а) (S) -tert-Butyl - ((3-chloro-8,8-dimethyl-7,8-dihydro-2Н-1,6,9-trioxa-9а-borabenzo [cd] αsulene -2yl) methyl) carbamate

A solution of tert-butyl - ((8,8-dimethyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methyl) carbamate (4.1 g, 12, 3 mmol) (example 9 (e)) and NCS (1.73 g, 13 mmol) in 100 ml of dichloroethane was stirred at 50 ° C for 5 hours.The solvent was evaporated at 40 ° C under reduced pressure and the residue was purified by chromatography on silica gel eluting with a mixture of ethyl acetate and petroleum ether (1:10) to give the title compound (2.6 g, 58%) as an oil. The racemic compound was separated by SFC (chiral column CHIRALPAK AD-H, elution with EtOH (20%) and CO ₂ (80%)) to give 1.2 g of the (S) -isomer (isomer eluting first, RT = 2.6 min ) and 1.2 g of the (R) isomer (second eluting isomer, RT = 3.5 min). LC / MS: 268.0 [M + H-100] +.

(b) Header Specified Compound

Dry HCl was bubbled through a solution of (S) -tert-butyl - ((3-chloro-8,8-dimethyl-7,8-dihydro-2H-1,6,9trioxa-9a-borabenzo [cd] αzulen-2-yl ) methyl) carbamate (1.2 g, 3.27 mmol) in diethyl ether (150 ml) at room temperature for 3 h and the mixture was then stirred for 18 h. The solvent was filtered and the filter cake was dried in vacuo to give ( S) -isomer (0.8 g, 80%) as a white solid. LC / MS: 268 [M + H] +. 1H NMR (400 MHz, DMSO-d6) δ 8.34 (brs, 3H), 7.52 (d, 1H, J = 8), 7.02 (d, 1H, J = 8), 5.58-5.56 (m, 1H), 4.42 -4.39 (m, 1H), 4.22-4.07 (m, 1H), 3.67-3.53 (m, 1H), 3.03-2.95 (m, 1H), 1.36-1.27 (ds, 6H).

Similarly, acid treatment with (R) -tert-butyl - ((3-chloro-8,8-dimethyl-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] αzulene-2- yl) methyl) carbamate gave the corresponding (R) -isomer (G25-CI (R)) as a white solid (1.2 g, 80%). LC / MS: 268 [M + H] +. 1H NMR (400 MHz,

- 25 038093

DMSO-d ₆ ) δ 8.33 (bs, 3H), 7.52 (d, 1H, J = 8), 7.02 (d, 1H, J = 8), 5.58 (m, 1H), 4.42-4.39 (m, 1H) , 4.21-4.07 (m, 1H), 3.67-3.54 (m, 1H), 3.03-2.95 (m, 1H), 1.36-1.27 (ds, 6H).

Example 13 (comparative). ((2S, 8R) -2- (aminomethyl) -3-fluoro-7,8-dihydro-2H-1,6,9-trioxa-9aborabenzo [cd] azulen-8-yl) methanol hydrochloride (C15-F)

(a) (S) -5 - ((2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorobenzaldehyde

A solution of 2-fluoro-5-hydroxybenzaldehyde (1.9 g, 13.6 mmol), (R) - (2,2-dimethyl-1,3-dioxolan-4yl) methyl 4-methylbenzenesulfonate (4.3 g, 15 mmol) and K ₂ CO ₃ (2.37 g, 17.2 mmol) in 40 ml of DMSO was stirred at 70 ° C for 16 h. Then the mixture was poured into 300 ml of water, extracted with ethyl acetate (200 ml), washed with water ( 200 ml) and brine (100 ml) and dried over anhydrous sodium sulfate. The solvent was evaporated at 40 ° C. under reduced pressure and the residue was purified by silica gel chromatography eluting with ethyl acetate and petroleum ether (1: 5) to give the product (2.9 g, 84%) as a colorless oil. LC / MS: 255.1 [M + H] +. 1H NMR (300 MHz, CD ₃ OD) δ 10.30 (s, 1H), 7.31-7.28 (m, 1H), 7.16-7.05 (m, 2H), 4.49-4.45 (m, 1H), 4.18-3.85 (m , 4H), 1.45-1.40 (d, 6H).

(b) (S) -1 - (5 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorophenyl) -2-nitroethanol

A mixture of copper (II) acetate (0.2 g, 1.1 mmol), (1R) -1,7,7-trimethyl-N- (pyridin-2-ylmethyl) bicyclo [2.2.1] heptan-2-amine (0.3 g, 1.23 mmol) (example 4, method B (b)) in ethanol (30 ml) was stirred at rt for 1 h, then a solution of (S) -5 - ((2,2- dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorobenzaldehyde (2.9 g, 11.4 mmol) in ethanol (50 ml). The reaction mixture was cooled to -35 to -40 ° C and then nitromethane (7 g, 115 mmol) was added dropwise keeping the temperature below -35 ° C, after which diisopropylethylamine (0.32 g, 2.50 mmol) was added ... The reaction mixture was stirred at -35 ° C for 24 h and then quenched with trifluoroacetic acid (0.29 g, 2.5 mmol). EtOAc (200 ml) was added to the resulting solution. The separated organic phase was washed with water (200 ml) and then concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with ethyl acetate and petroleum ether (1:10) to give the product (3.3 g, 92%) as a colorless oil. LC / MS: 316.1 [M + H] ⁺ .

(c) (S) -2-Amuho- 1 - (5 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorophenyl) ethanol

A solution of (S) -1 - (5 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorophenyl) -2-nitroethanol (3.2 g, 10 , 2 mmol) and Pd / C (10%, 0.5 g) in 70 ml of methanol was hydrogenated under a pressure of H ₂ 1 atm at room temperature for 48 h. Then it was filtered through a Celite pad and the filtrate was concentrated under reduced pressure to give crude product (2.9 g, 100%) as a colorless oil. It was used directly in the next step without further purification. LC / MS: 286.2 [M + H] ⁺ .

(d) (S) -2- (Dibenzylamino) -1- (5 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorophenyl) ethanol

Into a stirred solution of (S) -2-amino-1- (5 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2

- 26 038093 fluorophenyl) ethanol (2.9 g, 10.2 mmol) in 50 ml EtOH were added K ₂ CO ₃ (2.8 g, 20.3 mmol) and BnBr (3.6 g, 21 mmol). The reaction mixture was stirred overnight at room temperature. The solvent was removed under reduced pressure and the residue was purified by silica gel chromatography eluting with ethyl acetate and petroleum ether (1:10) to give the product (3.8 g, 80%) as a colorless oil. LC / MS: 466.2 [M + H] +.

(f) (S) -3 - ((Dibenzylamino) methyl) -7 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -4-fluorobenzo [c] [ 1,2] oxaborol-1 (3H) -ol

Into a solution of (S) -2- (dibenzylamino) -1- (5 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -2-fluorophenyl) ethanol (3, 3 g, 7.1 mmol) in dry toluene (40 ml) at −30 ° C. under N ₂ was added nBuLi (2.5 M in hexane, 20 ml, 50 mmol) dropwise over 30 min. After the addition, the mixture was stirred at 0 ° C for an additional 2 h and then cooled to -70 ° C, trimethyl borate (5.2 g, 50 mol) was added dropwise keeping the temperature below -50 ° C. After the addition, the reaction mixture was left to warm to 40 ° C for 3 h and then warmed to rt and stirred overnight. The reaction was quenched with 5% aqueous NaHCO ₃ solution (20 ml) and stirred vigorously for 15 min. The resulting suspension was filtered and the filtrate was separated. The organic layer was washed with water (20 ml x 3) and concentrated in vacuo to give the crude product (3 g, 86%) as a yellow oil. LC / MS: 492.2 [M + 1] +.

(f) Header Compound

A solution of (S) -3 - ((dibenzylamino) methyl) -7 - (((S) -2,2-dimethyl-1,3-dioxolan-4-yl) methoxy) -4 fluorobenzo [c] [1,2 ] oxaborol-1 (3H) -ol (3 g, 6.1 mmol) and Pd / C (10%, 0.7 g) in 50 ml of methanol with 2 ml of conc. HCl was hydrogenated at 1 atm H ₂ at room temperature for 48 hours. It was then filtered through a Celite pad and the filtrate was concentrated under reduced pressure to give an oil. The crude product was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient. The collected fraction was concentrated under reduced pressure. The residue was dissolved in diethyl ether (30 ml) and a saturated solution of HCl (g) in diethyl ether (30 ml) and the mixture was stirred at room temperature for 1 h. The solid was collected by filtration and washed with diethyl ether to give the title compound (0 , 4 g, 23%) as a white solid. LC / MS: 254.2 [M + H] +. 1H NMR (400 MHz, D2O) δ 7.20-7.16 (m, 1H), 6.94-6.91 (m, 1H), 5.55-5.53 (m, 1H), 4.17-4.04 (m, 3H), 3.70-3.62 (m , 3H), 3.193.14 (m, 1H).

Example 14 (comparative). ((2S, 8R or 2R, 8S) -2- (aminomethyl) -3-chloro-8-methyl-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] azulen-8-yl ) methanol (d6-Cl).

Example 15 (comparative). ((2S, 8S or 2R, 8R) -2- (aminomethyl) -3-chloro-8-methyl-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] azulen-8-yl ) methanol (G17-Cl)

(a) ((2-Methylallyloxy) methyl) benzene

A solution of metallic alcohol (80 g, 1.1 mol) in THF (100 ml) was added dropwise to a suspension of NaH (66 g, 1.65 mol) in THF (800 ml) at 25 ° C. under argon. After 1 h, a solution of benzyl bromide (207 g, 1.2 mol) in THF (100 ml) was slowly added and the reaction mixture was stirred at room temperature for 12 h. The reaction mixture was quenched with saturated NH ₄ Cl solution (200 ml) and extracted with ethyl acetate ( 3x200 ml). The combined organic layers were washed with water (100 ml) and brine (100 ml) and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure. The residue was distilled to obtain the target product (134 g, 74%) as a colorless oil. ¹ H NMR (400 MHz, CDCl3) δ 7.40-7.29 (m, 5H), 5.05 (s, 1H), 4.97 (s, 1H), 4.54 (s, 2H), 3.98 (s, 2H), 1.82 (s , 3H).

- 27 038093 (2- (Benzyloxymethyl) -2-methyloxirane

I m-CPBA, DCM q /

[JX ^ OBn ((2-Methylallyloxy) methyl) benzene (41.5 g, 256 mmol) was dissolved in DCM (1200 ml) and cooled to 0 ° C. M-CPBA (69.7 g, 384 mmol) was added and the mixture was stirred overnight at room temperature for 12 h. After filtering off the white precipitate, the filtrate was washed with saturated Na ₂ CO ₃ solution (200 ml), H ₂ O (200 ml ) and brine. After removal of the solvent under reduced pressure, the crude residue was purified by chromatography on silica gel, eluting with ethyl acetate and petroleum ether (1:20) to give the pure product (20 g, 44%) as a colorless oil. ¹ H NMR (400 MHz, CDCl3) δ 7.40-7.29 (m, 5H), 4.60 (q, J = 12.0 Hz, 2H), 3.61 (d, J = 11.0 Hz, 1H), 3 48 (d, J = 11.0 Hz, 1H), 2.78 (d, J = 4.9 Hz, 1H), 2.66 (d, J = 4.9 Hz, 1H), 1.43 (s, 3H).

3- (3- (Benzyloxy) -2-hydroxy-2-methylpropoxy) -2-bromobenzaldehyde

To a solution of (2- (benzyloxymethyl) -2-methyloxyrane (26 g, 145.9 mmol) in DMF (700 ml) was added K ₂ CO ₃ (42 g, 304.3 mmol), then 2-bromo-3- hydroxybenzaldehyde (30 g, 149.3 mmol) The suspension was stirred at 90 ° C for 6 h. The mixture, cooled to room temperature, was diluted with brine and extracted with ethyl acetate (200 ml × 3). The organic solvent was removed in vacuo and the residue was purified by chromatography on silica gel eluting with ethyl acetate and petroleum ether (1:20) to afford the pure product (27 g, 49%) as a light yellow oil.1H NMR (400 MHz, DMSO-d ₆ ) δ 10.29 (s, 1H), 7.512-7.41 (m, 3H), 7.31-7.23 (m, 5H), 4.91 (s, 1H), 4.53 (dd, J1 = 12.4 Hz, J ₂ = 17.2 Hz, 2H), 4.06 (d, J = 9.2 Hz, 1H), 3.91 (d, J = 9.2 Hz, 1H), 3.54 (d, J = 9.3 Hz, 1H), 3.47 (d, J = 9.3 Hz, 1H), 1.27 (s, 3H).

3- (3- (Benzyloxy) -2-hydroxy-2-methylpropoxy) -2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2yl) benzaldehyde

A solution of 3- (3- (benzyloxy) -2-hydroxy-2-methylpropoxy) -2-bromobenzaldehyde (21.3 g, 56.2 mmol), Pin ₂ B ₂ (28.6 g, 112.4 mmol), KOAc (6.1 g, 61.9 mmol), PdCl ₂ (dppf) DCM (1.23 g, 1.7 mmol) in DMF (150 ml) was degassed 3 times with nitrogen. The mixture was heated at 90 ° C for 16 h. After treatment of the reaction mixture with ethyl acetate and brine, the residue was purified by chromatography on silica gel, eluting with ethyl acetate and petroleum ether (1:20) to give the desired product (15.3 g, 64%) as light yellow oil. LC / MS: 367.1 [344 + Na] +.

(3- (Benzyloxy) -2-hydroxy-2-methylpropoxy) -3- (nitromethyl) -benzo [c] [1,2] oxaborol-1 (3H) -ol

Into an ice-cooled solution of 3- (3- (benzyloxy) -2-hydroxy-2-methylpropoxy) -2- (4,4,5,5tetramethyl-1,3,2-dioxaborolan-2-yl) benzaldehyde (18, 8 g, 44.1 mmol) in THF was added a solution of NaOH (1.76 g, 44.1 mmol) in water (100 ml). After stirring for 15 min, CH ₃ NO ₂ (3.3 g, 53 mmol) was added and the mixture was stirred at room temperature for 15 h. The reaction solution was acidified with AcOH to pH 3-5. The suspension was extracted with ethyl acetate (50 ml x 3). The combined organic layer was evaporated in vacuo and the residue was purified by chromatography on silica gel eluting with ethyl acetate and petroleum ether (1:10) to give the pure product (6.8 g, 40%) as a colorless oil. LC / MS: 386.0 [M-1] ^- .

(2- (Aminomethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-8-yl) methanol acetate

Pd (OH) 2 / C (200 mg) was added to a solution of 7- (3- (benzyloxy) -2-hydroxy-2-methylpropoxy) -3 (nitromethyl) benzo [c] [1,2] oxaborol-1 (3H ) -ol (1 g, crude) in AcOH (20 ml). The solution was degassed 3 times with H ₂ and stirred at room temperature for 12 hours. The reaction mixture was filtered through Celite and the filtrate concentrated in vacuo to give the crude product (1 g, crude) as a yellow solid.

tert-Butyl - ((8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulene-2yl) methyl) carbamate

- 28 038093

NaHCO ₃ (437 mg, 5 2 mmol) was added to the solution (2- (aminomethyl) -8-methyl-7,8-dihydro-2H-1,6,9trioxa-9a-borabenzo [cd] azulen-8-yl) methanol acetate (650 mg, 2.1 mmol) in t-BuOH (10 ml) and H ₂ O (10 ml) at room temperature. After stirring for 15 min, (Boc) ₂ O (854 mg, 3.9 mmol) was added and the reaction mixture was stirred at room temperature for 2 h. The mixture was acidified with AcOH to pH 6-7 and extracted with DCM (30 ml × 3) ... The combined organic layers were evaporated in vacuo, and the residue was purified by chromatography on silica gel, eluting with ethyl acetate and petroleum ether (1: 3), to obtain the target product (400 mg, 55%) as an oil (courses). LC / MS: 294.1 [M-55] ⁺ .

tert-Butyl - ((3-chloro-8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo ^] azulen-2-yl) methyl) carbamate

In a solution of tert-butyl - ((8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate ( 200 mg, 0.57 mol) in ACN (10 ml), NCS (77 mg, 0.57 mmol) was added and the solution was stirred at 90 ° С for 16 h. The reaction mixture was quenched with NH4Cl solution and extracted with ethyl acetate (20 ml × 3) ... The organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo. The crude residue was purified by chromatography on silica gel, eluting with ethyl acetate and petroleum ether (1: 3), to give the crude product (240 mg, crude) as a yellow oil. LC / MS: 284.1 [283 + H] ⁺ .

Connections specified in the header

Example 14 Example 16 tert-Butyl - ((3-chloro-8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2- yl) methyl) carbamate (240 mg, crude) was dissolved in a solution of TFA (1 ml) in DCM (10 ml). The solution was stirred at room temperature for 1 hour and then concentrated in vacuo.

The crude product was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient. The collected fraction was concentrated under reduced pressure to give the title compounds. ((2S, 8R or 2R, 8S) -2 (aminomethyl) -3-chloro-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-8- sludge) methanol.

LC / MS: 284.0 [M + H] ⁺ . 1H NMR (400 MHz, DMSO-d ₆ ) δ 8.23 (s, 3H), 7.52 (d, J = 8.6 Hz, 1H), 7.03 (d, J = 8.5 Hz, 1H), 5.56 (dd, J = 8.5 , 2.1 Hz, 1H), 4.55 (s, 1H), 4.15 (s, 1H), 3.59 (s, 1H), 3.45 (s, 2H), 3.04 (s, 1H), 1.21 (s, 3H). ((2S, 8S or 2R, 8R) -2- (aminomethyl) -3-chloro-8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-8 -yl) methanol. LC / MS: 284.1 [M + H] ⁺ . 1H NMR (400 MHz, DMSO-d6) δ 8.12 (s, 2H), 7.51 (s, 1H), 7.02 (s, 1H), 5.55 (s, 1H), 4.54 (s, 1H), 4.24-3.92 ( m, 1H), 3.78-3.29 (m, 3H), 3.02 (s, 1H), 1.25 (s, 3H).

Example 16 (comparative). ((2S, 8R, or 2R, 8S) -2- (aminomethyl) -3-bromo-8-methyl-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] azulene-8- yl) methanol (C18-Br).

Example 17 (comparative). ((2S, 8S, or 2R, 8R) -2- (aminomethyl) -3-bromo-8-methyl-7,8-dihydro-2H1,6,9-trioxa-9a-borabenzo [cd] azulene-8- yl) methanol (G19-Br), R + S1S + R1 f> ^OHone

00 ° 0С ° y ^ S + RIS + R1

Br ^ NH ₂ Br ^ νη ₂

Example 16 Example 17 tert-Butyl - ((3-bromo-8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulene-2- yl) methyl) carbamate

In a solution of tert-butyl - ((8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9а-borabenzo [cd] azulen-2-yl) methyl) carbamate ( 200 mg, 0.57 mmol) in ACN (10 ml), NBS (102 mg, 0.57 mmol) was added and the solution was stirred at 90 ° C for 1 h. The reaction mixture was quenched with NH ₄ Cl solution and extracted with ethyl acetate (20 mlx3). The organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo. The crude residue was purified by chromatography on silica gel, eluting with ethyl acetate and petroleum ether (1: 3), to give the product (230 mg, crude) as a pale yellow solid. LC / MS: 328.1 [M-Boc + H] ⁺ .

- 29 038093

Connections specified in the header

Example 15 Example 17 tert-Butyl - ((3-bromo-8- (hydroxymethyl) -8-methyl-7,8-dihydro-2H-1,6,9-trioxa-9a-borabenzo ^] azulen-2-yl ) methyl) carbamate (230 mg, crude) was dissolved in a solution of TFA (1 ml) in DCM (10 ml). The solution was stirred at room temperature for 1 hour and then concentrated in vacuo. The crude product was purified by preparative HPLC using a Daisogel 10μ C18 column (250x50 mm) and eluting with a water / acetonitrile (0.05% TFA) gradient. The collected fraction was concentrated under reduced pressure to afford the title compounds ((2S, 8R or 2R, 8S) -2 (aminomethyl) -3-bromo-8-methyl-7,8-dihydro-2H-1,6,9- trioxa-9a-borabenzo [cd] azulen-8-yl) methanol LC / MS 328.0 [M + H] +. ¹ H NMR (400 MHz, DMSO-d6) δ 8.10 (s, 3H), 7.65 (d, J = 8.3 Hz, 1H), 7 07-6 88 (m, 1H), 5.56-5.39 (m, 1H) , 5.36-5.17 (m, 1H), 4.61-4.52 (m, 1H), 4.19-4.07 (m, 1H), 3.62 (d, J = 11.9 Hz, 1H), 3.513.39 (m, 2H), 3.04 (s, 1H), 1.18 (s, 3H). ((2S, 8S, or 2R, 8R) -2- (aminomethyl) -3-bromo-8-methyl-7,8-dihydro2H-1,6,9-mriokсα-9α-boronαbenzo [cd] azulene-8- yl) meta-nol LC / MS 328.0 [M + H] ⁺ . 'H NMR (400 MHz, DMSOd6) δ 8.13 (s, 2H), 7.65 (s, 1H), 6.98 (s, 1H), 5.47 (s, 1H), 5.26-5.06 (m, 1H), 4.53 (s , 1H), 4.19-3.97 (m, 1H), 3.83-3.56 (m, 1H), 3.51-3.26 (m, 2H), 3.01-2.93 (m, 1H), 1.25 (s, 3H).

In vitro assays.

Example 18. Determination of MIC against mycobacteria.

The measurement of the minimum inhibitory concentration (MIC) against M. tuberculosis strains for each compound tested was performed in 96-well flat-bottomed polystyrene microtiter plates in a final volume of 100 μl. Ten 2-fold dilutions were performed in undiluted DMSO, starting at 50 mM. Drug solutions were added to Middlebrook 7H9 medium (Difco) and isoniazid (INH) (Sigma Aldrich) was used as a positive control with 2-fold dilutions of INH starting at 160 μg / ml. The inoculum was standardized to approximately 1 x 10 ⁷ CFU / ml and diluted 1 in 100 in Middlebrook 7H9 broth (Difco). This inoculum (100 μl) was added to all wells of the plate, but wells G-12 and H-12 were used as blind controls. All plates were placed in a sealed box to prevent drying out of the peripheral wells and incubated at 37 ° C without shaking for six days. A resazurin solution was prepared by dissolving one resazurin tablet (Resazurin Milk Testing Tablet, Ref 330884Y 'VWR International Ltd) in 30 ml sterile PBS (phosphate buffered saline). 25 μl of this solution was added to each well. Fluorescence was measured (Spectramax M5 Molecular Devices, excitation wavelength 530 nm, emission wavelength 590 nm) after 48 hours to determine the MIC value.

Example 19. MIC against clinical strains.

The VASTEC MGIT 960 System (Becton Dickinson) was used to perform MIC determination in clinical isolates (Institute Carlos III) following the manufacturer's instructions. The resistance pattern of clinical isolates is indicated by the following abbreviations: H - isoniazid, R - rifampicin, T - ethionamide, S - streptomycin, E - ethambutol, Z - pyrazinamide, K - kanamycin, A - amikacin, and CP - capreomycin. The results for the compound of example 4 G4-Cl are shown in table. 1A, 1B, 2A and 2B and in FIG. 3 and 4. The results for the compound of example 2 G2-Br are shown in table. 2C and 2D and in FIG. 4.

Table 1

MIC values for Example 4 compound G4-Cl tested against susceptible (A) and resistant (B) clinical isolates of M. tuberculosis A

Strain 362 457 3 356 357 370 137 169 192 199 206 207 208223 ~

MIC 0.04 0.04 0.04 0.04 0.04 0.04 0.08 0.08 0.08 0.08 0.08 0.08 0.080.08 | (μM) ____________________________________________________________________ Strain 231 237 247 248 249 250 253 255 256 257 261 265 269281

MIC 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.080.08 (μM) ____________________________________________________________________ Strain 292 296 311 314 316 317 322 323 324 326 327 328 329332

MIC 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.080.08 (μM) ____________________________________________________________________ Strain 333 337 358 361 371 385 391 424 440 442 460 481 716729

MIC 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.080.08 (μM) ____________________________________________________________________ Strain 730 731 733 734 736 737 52 267 374 274 325 705161

MIC 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.16 0.16 0.160.31 (μM)

- 30 038093

Strain 1819 670 330 198 242 409 141 415 330 Sustainability HSRZ HRZ RSR H H HR HRT HRT Hs MIC (μM) <0.02 0.04 0.04 0.08 0.08 0.08 0.08 0.08 0.08 Strain 175 709 732 201 202 277 605 123 106 Sustainability S S S Hre S Η HSERZ HSERZ HR ACp KTACp MIC (μM) 0.08 0.08 0.08 0.08 0.08 0.08 0.16 0.16 0.16 Strain 562 139 514 1672 167 254 297 192 CR Sustainability HSERZ HSERZK HSRT HSRZ HSRZ Η HSR Hs HSERZ KTA TASr MIC (μM) 0.08 0.08 0.08 0.08 0.08 0.16 0.16 0.31 0.31

FIG. 3 is a graphical representation of the MIC data shown in table. 1A and 1B for the compound of Example 4 G4-Cl, expressed as the number of strains with a particular MIC (y) value versus a particular MIC (x) value obtained in μM. As can be seen in FIG. 3, G4-Cl (the compound of example 4) showed an MIC value of less than 1 μM against more than 85 clinical isolates of 97 strains tested (susceptible and resistant), indicating good activity of this compound against a significant number of clinical isolates of M. tuberculosis strains. The assay showed a measured MIC <0.2 μM versus 1 strain, measured MIC 0.04 μM versus 8 strains, measured MIC 0.08 μM versus 76 strains, measured MIC 0.16 μM versus 8 strains, and measured MIC 0.31 μM versus 3 strains.

Tables 2A and 2B

MIC values for Example 4 compound G4-Cl tested against susceptible (A) and resistant (B) clinical isolates of M. tuberculosis A

Strain 137 169 192 199 206 207 208 223 231 237 247 248 MIC (mkM) 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 Strain 249 255 261 265 269 281 292 314 316 317 322 323 MIC (mkM) 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 Strain 324 326 327 328 329 332 333 358 MIC (mkM) 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 I 3 Strain Resilience MIC (μM) Strain Sustainability MIC (μM) 175 198 242 330 141 S Η H HSR HRT 0.08 0.08 0.08 0.08 0.08 1819 1672 167 139 123 HSRZ HSRZ HSRZ HSERZKTACp HSERZKTACp 0.02 0.08 0.08 0.08 0.16

Tables 2C and 2D

MIC values for Example 2 compound G2-Br tested against susceptible (A) and resistant (B) clinical isolates of M. tuberculosis C

Strain 137 169 192 199 206 207 208 223 231 237 247 248 MIC (mkM) 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 Strain 249 255 261 265 269 281 292 314 316 317 322 323 MIC (mkM) 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08 Strain 324 326 327 328 329 332 333 358 MIC (mkM) 0.08 0.08 0.08 0.08 0.08 0.08 0.08 0.08

D

Strain Sustainability MIC (mkM) 175 S 0.08 198 H 0.08 242 H 0.08 330 141 HSR HRT 0.08 0.08 Strain 1819 1672 167 139 123 Sustainability HSRZ HSRZ HSRZ HSERZKTACp HSERZKTACp MIC (mkM) 0.04 0.08 0.08 0.08 0.08

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FIG. 4 is a graphical representation of the MIC data shown in table. 2A-2D for compound G2-Br (compound of example 2, open bar) and compound G4-Cl (compound of example 4, dark bar), expressed as the number of strains with a specific MIC value (y) versus a specific MIC value obtained (x) in μM. As can be seen in FIG. 4, G4-Cl (compound of Example 4) and G2-Br (compound of Example 2) showed an MIC value of less than 1 μM against all but 1 of the 40 clinical isolates of M. tuberculosis strains tested in this experiment. The analysis showed a measured MIC <0.2 μM against 1 strain (compound of example 4), measured MIC 0.04 μM against 1 strain (compound of example 2), measured MIC 0.08 μM against 40 strains (compound of example 2 and compound of example 4 ) measured MIC 0.16 μM against 1 strain (compound of example 2 and compound of example 4), and MIC 0.31 μM, measured neither for the compound of example 2 nor for the compound of example 4 against any strain.

Example 20. Analysis of general antibacterial activity.

Antibacterial activity on whole cells was determined by broth microdilution using the method recommended by the Clinical and Laboratory Standards Institute (CLSI) Document M7-A7, Methods for Dilution Susceptibility Tests for Bacteria that Grow Aerobically. ).

Table 3 shows the MIC values against bacterial strains K12, E. coli K12 tolC / Tn10, A. baumannii ATCC 17978, and P. aeruginosa PA01 for the compounds disclosed in the examples. As you can see, the compounds from the examples in most cases do not have significant activity against several pathogenic gram-negative bacteria, as well as efflux pump-deficient E. coli. But, as shown in table. 4 below, the compounds disclosed in the examples have significant activity against M. tuberculosis. Moreover, as can be seen, comparative tricyclic benzoxaborols lacking 4-halogen (for example, C2-H, C5-H and C12-H) have a higher activity against these bacterial strains, and tricyclic benzoxaborole compounds with a third ring, which is a seven-membered ring between the 1 and 7 positions of benzoxaborole, additionally having substitution with 4-halogen, 3-aminomethyl with (S) stereochemistry at the 3 position (eg G2-Br and G4-Cl) have very poor activity against these bacteria. This contrasts markedly with their respective activities against M. tuberculosis, where 4-halogen compounds usually show very good activity, but tricyclic benzoxaborols without 4-halogen are inferior (compare M. tuberculosis MIC values for such a set of compounds in Tables 4A and 4B) ...

Table 3

MIC values against non-mycobacterial _____ strains for compounds of formula II or IIa____

Compound MIC: E. coli K12 [μg / ml] MIC: E. coli K12 Y1C :: Tn10 [μg / ml] MIC: A. baumannii ATCC 17978 [μg / ml] MIC: R. aeruginosa PA01 [μg / ml] Example 1 Gl-Br > 64 > 64 > 64 > 64 Example 2 G2-Br 64 64 64 64 Example 3 G3-C1 > 64 > 64 > 64 > 64 Example 4 G4-C1 64 64 64 64 Example 5 G5-F 32 64 > 64 4 Example 6 G6-I - - - - Example 7 G7-C1 > 64 > 64 > 64 > 64 Example 8 G8-Br > 64 > 64 > 64 > 64 Example 9 G9-Br > 64 > 64 > 64 > 64 Example 10 G10-Br > 64 > 64 > 64 > 64 Example 11 G11-C1 > 64 > 64 > 64 > 64 Example 12 G12-C1 > 64 > 64 > 64 > 64 C1-H - - - - C2-H 2 4 2 2

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sz-n - - - - C4-Bg 64 64 64 64 C5-H - - - - C6-C1 64 64 64 64 C7-C12 - - - - C8-C1 - - - - C9-C1 - - - - S10-H - - - - C11-H 2 2 4 2 C12-H 4 2 4 16 С13-С1 - - - - C14-C12 - - - - C15-F - - - - C16-C1 GSK3 309930А AN12471 01 - - - - С17-С1 GSK3309934A AN12470.01 - - - - C18-Bg GSK3337512A AN12344 01 - - - - C19-Bg GSK3309932A AN12343 01 - - - -

For biochemical assays, N-terminal six-histidine-tagged LeuRS were overexpressed in Eschenchia coli, which were codon-optimized E. coli (GenScript, Piscataway NJ, USA), from human mitochondria and cytoplasm, and M. tuberculosis. The N-terminal six-histidine-tagged LeuRS proteins were overexpressed and purified according to Novagen (Madison, WI, USA) using an overexpressed E. coli strain BL21 (DE3) carrying the T7 RNA polymerase gene.

Example 22. Assay of aminoacylation.

Experiments were performed in 96-well microtiter plates using 80 μl reaction mixtures containing 50 mM HEPES-KOH (pH 8.0), 30 mM MgCl ₂ , 30 mM KCl, 13 μM L [ ¹⁴ C] leucine (306 mCi / mmol , Perkin-Elmer), 15 μM total E coli tRNA (transport RNA, tRNA) (Roche, Switzerland), 0.02% (w / v) BSA (bovine serum albumin), 1 mM DTT, 0.2 pM LeuRS and 4 mM ATP (adenosine triphosphate) at 30 ° C. Reactions were started by the addition of 4 mM ATP. After 7 min, the reactions were quenched and tRNA was precipitated by the addition of 50 μl of 10% (w / v). TCA (trichloroacetic acid) and transferred to 96-well nitrocellulose membrane filter plates (Millipore Multiscreen HTS, MSHAN4B50). Each well was then washed three times with 100 μl of 5% TCA. The filter plates were then dried under a heating lamp and the precipitated L- [ ¹⁴ C] leucine tRNALeu was quantified by liquid scintillation counting using a Wallac MicroBeta Trilux liquid scintillation counter, model 1450 (PerkinElmer, Waltham, MA, USA). The only difference was with human cytoplasmic LeuRS when using tRNA isolated from brewer's yeast (Roche Diagnostics GmbH)

Example 23. Determination of IC ₅₀ .

To determine the inhibitory concentration that reduces the enzyme activity by 50% (IC ₅₀ ), increasing concentrations of the compound (Anacor Pharmaceuticals Inc, Palo Alto, CA, USA) were incubated with LeuRS enzyme, tRNA and L-leucine for 20 minutes. Reactions were initiated by the addition of 4 mM ATP. The reactions were stopped after 7 min, then precipitated and the radioactivity was counted. _{IC 50} values were determined using the Graphpad Prism software package (Graphpad Software Inc (La Jolla, CA, USA).

Example 24 HepG2 Cytotoxicity Assay.

HepG2 cells (HB-8065) were plated in fresh medium (Eagle's Minimum Essential Medium (EMEM) supplemented with 5% fetal bovine serum and 2 mM L-glutamine) one day prior to subculture of the plates. On the day of plating the plates, a cell suspension of 100,000 cells / ml was prepared in culture medium. The cell suspension (100 μl) was added to each well of a black 96-well clear-bottomed microplate coated with collagen (Becton Dickinson), except for the wells of column 11, which received 100 μl of culture medium only. The plates were incubated for 24 hours. A 10 dose range of test substances was created by making serial 1: 2 dilutions from a stock solution in 100% DMSO and a 1: 200 dilution of each dose in medium was made to a final concentration of 0.5% DMSO. After 24 h, the culture medium was removed from the plate, and 150 μl of dilutions of the test compounds were added in duplicate and 150

- 33 038093 μl 0.5% DMSO in culture medium in the wells of columns 11 and 12 (blind control). The plates were incubated for 48 hours at 37 ° C, 5% CO ₂ , at a relative humidity of 95%. The medium was then removed and 200 μl of fresh culture medium and 50 μl of resazurin solution were added to each well and incubated for 1.5 hours. The plates were removed from the incubator and the fluorescence was stabilized at room temperature in the dark for 15 minutes. Resazurin (BDH) was used to determine cell viability. Resazurin has been used as a redox indicator that gives a colorimetric change and a fluorescent signal in response to metabolic activity. As cells grow, metabolic activity induces chemical reduction of resazurin, indicated by a color change from a non-fluorescent blue to a reduced fluorescent pink form. The degree of fluorescence of resazurin is therefore an indicator of the number of viable cells in the culture system. Fluorescence was measured at an excitation wavelength of 515 nm and an emission wavelength of 590 nm in a 1420 Multilabel HTS microplate reader, Victor 2, (Wallac).

The fluorescence value of each well was corrected by subtracting the background value (mean value for wells in column 11) from the absolute value. Percentage inhibition was calculated relative to DMSO control wells (mean for wells in column 12). For each compound, the mean of the duplicate samples was calculated and a sigmoidal dose response curve (variable slope) was constructed, which was flattened by non-linear regression (GraphPad) to calculate the IC ₅₀ (Tox50).

Example 25. Effect of the compounds described herein against Mycobacterium tuberculosis.

The compounds of the present invention were tested for antibacterial activity against the species Mycobacterium tuberculosis, and were also tested for toxicity to human liver cells HepG2. Exemplary compounds of the invention were compared to comparative compounds Ci-Η at C19-Br as shown in Table II. 4A and 4B.

Table 4A

_{IC 50} values for LeuRS inhibition, MIC values against the standard M. tuberculosis strain Mtb H37Rv, toxicity values for human HepG2 cells, and selectivity values for some comparative tricyclic benzoxaborol compounds

Cells

Connection structure

Connection designation

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C9-C1 С% Cl ^ nh ₂ 37.59 - 5.0 > 50 > 10 C10-H 0 Z > 300 - - > 5.0 > 50 ten Cll-H Z 0.51 - - 1.56 > 50 (40%) > 32 C12-H with with? n% n ₂ 1.33 - - > 5.0 24.5 > 4.9 C13-C1 Λ C ”' ⁰ 1 2.16 5.0 > 50 > 10 C14-C12 ^{with |} Dee Cl 4.67 > 5.0 > 50 > 10 EXAMPLE 13 C15-F ((2S, 8R) -2 (aminomethyl) -3fluoro-7,8dihydro2H-1,6,9trioxa-9aborabenzo [c <1] azulene-8yl) methanol χ- OH pi γ g hci F ~~ NH ₂ 0.48 - - 0.55 > 50 > 10 EXAMPLE 14 C16-C1 ((8R) -2 (aminomethyl) -3chloro-8methyl-7,8dihydro2H-1,6,9trioxa-9aborabenzo [c <1] azulene-8yl) methanol EXAMPLE 15 С17-С1 ((8S) -2 (aminomethyl) -3chloro-8methyl-7,8dihydro2Н-1,6,9trioxa-9abora6eH3o [cd] azulene-8yl) methanol k0 ^HO nh ₂ nh ₂ 4.17 3.13 - 1.25 0.93 > 50 > 50 > 4 > 50

- 35 038093

EXAMPLE 16 C18-Br ((8R) -2 (aminomethyl) -3 bromo-8methyl-7,8dihydro2H-1,6,9trioxa-9aborabenzo [cc!] Azulene-8yl) methanol -I nh ₂ 2.69 - - 1.25 > 50 > 40 EXAMPLE 17 C19-Bg ((8S) -2 (aminomethyl) -3 bromo-8methyl-7,8dihydro2H-1,6,9trioxa-9aborabenzo [c <1] azulene-8yl) methanol nh ₂ 1.97 - - 0.925 > 50 > 50

Table 4B

_{IC 50} values for LeuRS inhibition, MIC values against the standard M. tuberculosis strain Mtb H37Rv, toxicity values for human HepG2 cells, and selectivity values for compounds of formula II or formula IIa

Connection designation Connection structure Mtb LeuRS IC50 (μM) human cyto LeuRS IC50 (μM) Human mito LeuRS IC50 (μM) Mtb H37Rv MIC (μM) (V) HepG2 cells 48 h Tox50 (μM) (A) Selectivity index (A / B) EXAMPLE 1 Gl-Br 0.154 0.18 > 50 > 277 EXAMPLE 2 G2-Br ¢ 4 Br ^ NH ₂ 0.115 118 > 300 0.07 292.4 4177 EXAMPLE 3 G3-Br "I AM CI 0.244 0.47 > 50 > 106 EXAMPLE 4 G4-C1 О ¢ 6 Cl 'NH ₂ 0.148 94.7 > 300 0.08 > 1000 > 12500 EXAMPLE 5 G5-F Z 0.46 0.6 49.1 164

- 36 038093

As you can see from the table. 4B, the compounds of Examples 2, 4, 10 and 12 (G2-Br, G4-C1, G10-Br and G12-C1) show increased selectivity for inhibiting the growth of M. tuberculosis compared to toxicity to human HepG2 cells for the tricyclic benzoxaborol compound. in the third ring, which is a seven-membered ring between positions 1 and 7 of benzoxaborole, additionally having substituents 4-halogen and 3-aminomethyl with (S) stereochemistry at position 3.

Table 4A and 4B show a comparison of some tricyclic benzoxaborole compounds with and without a halogen substituent, some tricyclic benzoxaborol compounds with and without a halogen substituent at the 4-position of the benzoxaborole ring structure, and some bicyclic compounds. From the MIC values for Mtb H37Rv (B) and the Tox ₅₀ values for HepG2 cells 48 h (A), it is possible to determine the selectivity for inhibition of M. tuberculosis relative to the inhibitory effect (toxic effect) on human cells for these compounds (see the far right column in table. 4A and 4B)

The compounds of Example 2 G2-Br and Example 4 G4-Cl were found to have selectivity indices for M. tuberculosis of 4177 and> 12500, respectively (see Table 4B). Further, as can be seen from the table. 4B, the IC ₅₀ values for these compounds against M. tuberculosis were found to be submicromolar, 0.3 and 0.1, respectively. As can be seen, the selectivity index (SI) of the compounds of Example 2 G2-Br and Example 4 G4-Cl towards M. tuberculosis is surprisingly increased compared to other benzoxaborol compounds. The compounds of Example 2 G2-Br and Example 4 G4-Cl, which are tricyclic benzoxaborole compounds having a halogen substituent at the C-4 position of the benzoxaborole ring and an aminomethyl substituent at the C3 position of the benzoxaborole ring having (S) relative stereochemistry at this stereocenter, surprisingly are more selective for activity against M. tuberculosis than other benzoxaborol compounds lacking some of these features regarding the inhibitory effect (toxic effect) on human cells for these compounds. In addition, the MIC values against M. tuberculosis strain H37Rv for the compounds of example 2 G2-Br and example 4 G4-Cl are <0.1 μM, in contrast to other benzoxaborol compounds in this study.

Thus, as can be seen from the table. 4B, the compounds of Example 2 G2-Br and Example 4 G4-Cl have an SI for Mycobacterium tuberculosis of 4177 (Example 2 G2-Br) and> 12500 (Example 4 G4-Cl), respectively. These SI values are surprisingly better than any of the comparative compounds tested to date.

The addition of a chloro or bromo substituent at the C4 position of the benzoxaborole ring imparts an unexpected increase in the selectivity index. C2-H (racemic compound, no halogen substituent at the C4 position of the benzoxaborole ring) has a selectivity index> 26, while the compound of example 1 G1-Br (racemic, bromo substituent at the C-4 position of the benzoxaborole

- 37 038093 rings) has SI> 277. Similarly, Example 3 G3-C1 (racemic, chloro substituent at the C-4 position of the benzoxaborole ring) has an SI> 106 compared to C2-H with SI> 26.

The formation of a third ring involving the 1 and 7 positions of the benzoxaborole ring confers an unexpected increase in the selectivity index. C4-Br, the (S) enantiomer of the comparative non-tricyclic benzoxaborol compound with Br at the C4 position of the benzoxaborole ring, has SI 320, while the compound of Example 2 G2-Br, (S) the enantiomer of tricyclic benzoxaborol with Br at C-4 position, has SI 4177 Similarly, C6-C1, (S) the enantiomer of the non-tricyclic benzoxaborol reference compound with C1 at the C4 position of the benzoxaborole ring, has SI 363, while Example 4a is G4-C1, (S) the enantiomer of tricyclic benzoxaborol with Cl at the C-4 position, has SI> 12500.

If we compare the SI of the compound of example 2 G2-Br and the compound of example 4 G4-Cl c SI of the compound C5H, the (S) enantiomer of the non-tricyclic reference benzoxaborol compound with H at the C4 position of the benzoxaborole ring, it can be seen that the SI of such a compound without a halogen substituent at the position The C4 is only 3, indicating that such a compound has very little selectivity for inhibiting M. tuberculosis from killing human cells.

Certain substitutions on the 7-membered tricyclic ring impart an unexpected increase in the selectivity index. Table 4B shows the compound of example 9 G9-Br and the compound of example 11 G11-Cl with indices SI> 167 and> 185, respectively, while comparative compounds C9-a (tricyclic benzoxaborole with a chloro substituent at position C4 and -CH ₃ on R ³ and R ^{4 of the} 7-membered ring) and C10-H (tricyclic benzoxaborole with hydrogen at the C4 position and —CH ₃ on R ³ and R ^{4 of the} 7-membered ring) have indices SI 10. This most likely indicates that this substitution in positions R ³ and R ⁴ does not contribute to the selectivity for M. tuberculosis compared to the inhibitory effect (toxic effect) on human cells for these compounds. This also suggests that the presence of halogen substitution at the C4 position of the benzoxaborole ring (see C9-a) is insufficient to weaken the negative effect of methyl substitution both at the R ³ position and at the R position of the 7-membered tricyclic ring at the R3 / R ^{4 position.} ...

In other respects, the compounds of Example 2 G2-Br and Example 4 G4-Cl also have SI values surprisingly higher than related ring-opened benzoxaborols (substituted benzoxaborols) having no halogen substituent at the C4 position of the benzoxaborole ring. Compare the SI for C5H (5) with the SI for the compounds of Example 2 G2-Br and Example 4 G4-Cl. Benzoxaborols that are not tricyclic benzoxaborols, but which have a halogen at the C4 position of the benzoxaborole ring, show increased SI values compared to no halogen, but still show SI values significantly lower than SI values for the compounds of Example 2 G2-Br and Example 4. G4-Cl (compare C5-H with C3-Br and C6-a, but then compare all three compounds C5-H, C3-Br and C6-a with the SI values of Example 2 G2-Br and Example 4 G4-Cl) ...

Thus, the tricyclic benzoxaborols of the invention, in particular the compounds of Example 2 G2-Br and Example 4 G4-Cl, exhibit unexpectedly higher SI values compared to the SI values of related benzoxaborols for M. tuberculosis relative to human cells.

It should be borne in mind that the invention encompasses all combinations of aspects with all other suitable aspects and / or illustrative embodiments described herein. in this document.

It is understood that the examples and embodiments described in this document are presented for illustrative purposes only, and it is assumed that various modifications or changes in their light, which will be proposed by specialists in this field, are within the spirit and scope of this application and are included in the scope of the claims according to the appended claims. All publications, patents, and patent applications cited in this specification are incorporated by reference in their entirety for all purposes.

Claims (13)

CLAIM 1. A method of treating a disease caused by an infection of the Mycobacterium tuberculosis complex in an animal, comprising administering to an animal in need of such treatment an effective amount of a compound representing (S) - (3-chloro-7,8-dihydro-2H-1,6 , 9-trioxa-9a-borabenzo [cd] α-zulene-2yl) methanamine, or a pharmaceutically acceptable salt thereof. 2. The method of claim 1, wherein the pharmaceutically acceptable salt is ((S) - (3-chloro-7,8dihydro-2H-1,6,9-trioxa-9a-borabenzo [cd] azulen-2-yl) methanamine dihydrosulfate-H ₂ O. 3. The method of claim 1, wherein the compound is administered as a pharmaceutical composition with at least one pharmaceutically acceptable excipient. 4. The method of claim 1, wherein the compound is administered in combination with a second therapeutic agent that is not the compound or a pharmaceutically acceptable salt thereof according to claim 1. 5. The method of claim 4, wherein the second therapeutic agent is independently selected from isoniazid, rifampin, - 38 038093 pyrazinamide, ethambutol, moxifloxacin, rifapentin, clofazimine, bedaquiline (TMC207), nitroimidazooxazine RA-824, delamanide (ORS-67683), oxazolidinone, an analogue of EMB (ethambutol) SQ109 and benzothirobinide. 6. The method of claim 5, wherein the oxazolidinone is linezolid, tedizolid, radezolid, sutezolid (PNU-100480) or posizolid (AZD-5847). 7. The method of claim 4, wherein the second therapeutic agent is an antiviral agent. 8. The method of claim 7, wherein the antiviral agent is an antiretroviral agent. 9. The method according to claim 8, wherein the antiretroviral agent is zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxil, fozivudine, todoxil, emtricitabine, alovudine, amdoxovir, elviridinitabine, efvirudinaval , oltipraz, cappavirine, lersivirine, GSK2248761, TMS-278, TMS-125, etravirine, saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, brecanavir, darunavir, atazanavir, lavinavir-tipranavir, TMC-20 -1249, PRO-542, PRO-140, TNX-355, BMS-806, BMS-663068 and BMS626529, 5-Helix, raltegravir, elvitegravir, GSK1349572, GSK1265744, vicrivirok (Sch-C), Sch-D, TAK779, maraviroc, TAK449, didanosine, tenofovir, lopinavir, or darunavir. 10. The method of claim 4, wherein the second therapeutic agent is a therapeutic agent approved or recommended for the treatment of tuberculosis. 11. The method of claim 4, wherein the combination comprises a pharmaceutically acceptable excipient, adjuvant, or diluent. 12. The method of claim 1, wherein the disease is tuberculosis. 13. The method of claim 1, wherein the animal is a human.