Aminoglycoside derivatives and their use in treatment of genetic disorders

Abstract

The present invention provides novel aminoglycosides (e.g., compounds of formulae A, B, I, I *, III, or III *, including compounds represented by formulae Ia, I * *, I * a, I * b, IIIa, III * *, III * a, and III * b) designed to exhibit termination codon mutation readout activity represented by the formulae defined in the specification. The invention also provides pharmaceutical compositions comprising the novel aminoglycosides, and their use in the treatment of genetic diseases and disorders, such as diseases and disorders associated with termination codon mutations.

Description

[0001] This application is a divisional application of the Chinese national phase patent application with application number 201880051071.0 filed on June 5, 2018 and entitled “Aminoglycoside derivatives and their use in the treatment of genetic disorders”.

[0002] FIELD AND BACKGROUND OF THE INVENTION

[0003] The present invention, in some embodiments thereof, relates to aminoglycosides, and more particularly, but not exclusively, to novel aminoglycoside derivatives and their use in increasing the expression of a gene having a stop codon mutation and / or in treating a genetic disorder.

[0004] Many human genetic disorders are caused by nonsense mutations, in which one of the three stop codons (UAA, UAG or UGA) replaces the amino acid coding codon, resulting in premature termination of translation and ultimately a truncated inactive protein. Currently, hundreds of such nonsense mutations are known, and several have been shown to be the cause of cases of certain fatal diseases, including, for example, cystic fibrosis (CF), nephrotic cystinosis, Duchenne muscular dystrophy (DMD), ataxia-telangiectasia, Hurler syndrome, hemophilia A, hemophilia B, Tay-Sachs, Rett syndrome, Usher syndrome, severe epidermolysis bullosa, etc. For many of these diseases, there is currently no effective treatment.

[0005] Some aminoglycoside compounds have been shown to have therapeutic value in the treatment of various genetic diseases because they are able to induce ribosome readout stop codon mutations, producing full-length proteins from a portion of the mRNA molecule.

[0006] Aminoglycosides are highly potent broad-spectrum antibiotics that are commonly used to treat life-threatening infections. It is generally believed that the mechanism of action of aminoglycoside antibiotics, such as paromomycin (see Figure 1), involves interaction with prokaryotic ribosomes, and more specifically, involves binding to the decoding A-site of 16S ribosomal RNA, resulting in inhibition of protein translation and interference with translation fidelity.

[0007] Several results of bacterial ribosome structure determination and crystallographic and NMR structures of bacterial A-site oligonucleotide models have provided useful information for understanding the decoding mechanism in prokaryotic cells and how aminoglycosides cause deleterious misreading of the genetic code. These and other studies have proposed the hypothesis that the affinity of the A-site for noncognate mRNA-tRNA complexes increases upon aminoglycoside binding, thereby preventing the ribosome from effectively discriminating between noncognate and cognate complexes.

[0008] The enhancement of termination inhibition by aminoglycosides in eukaryotes is thought to occur by a mechanism similar to the activity of aminoglycosides in prokaryotes to interfere with translation fidelity during protein synthesis, i.e., binding of specific aminoglycosides to the ribosomal A site may induce conformational changes that stabilize near-cognate mRNA-tRNA complexes rather than inserting release factors. Aminoglycosides have been shown to inhibit various stop codons with significantly different efficiencies (UGA>UAG>UAA), and the inhibitory effect was found to also depend on the identity of the fourth nucleotide immediately downstream of the stop codon (C>U>A≥G) and the local sequence context surrounding the stop codon.

[0009] Desirable features of an effective readout drug are oral administration and minimal or no effect on bacteria. As with any unnecessary use of antibiotics, antibacterial activity of a readout drug is undesirable, especially for the gastrointestinal (GI) biota, as it can have adverse effects by disturbing the GI biota balance and developing resistance. In this regard, in addition to the above limitations, most clinical aminoglycosides are extremely selective for bacterial ribosomes and do not significantly affect the cytoplasmic ribosomes of human cells.

[0010] To circumvent the above limitations, the biopharmaceutical industry is seeking new stop codon mutation suppressors by screening large chemical libraries for nonsense readout activity.

[0011] The first experiments to demonstrate the aminoglycoside-mediated inhibition of the cystic fibrosis transmembrane conductance regulator (CFTR) stop codon mutation demonstrated that the premature stop codon mutation found in the CFTR gene can be inhibited by members of the gentamicin family as well as (G-418) (see Figure 1), as measured by the appearance of full-length functional CFTR in a human bronchial epithelial cell line.

[0012] Inhibition experiments with intestinal tissue from CFTR- / - transgenic mouse mutants carrying the human CFTR-G542X transgene have shown that treatment with gentamicin and, to a lesser extent, tobramycin has resulted in the appearance of human CFTR protein in the glands of treated mice. Most importantly, clinical studies using double-blind, placebo-controlled, crossover trials have shown that gentamicin can inhibit stop codon mutations in affected patients, and gentamicin treatment improved transmembrane conductance in the nasal mucosa of 19 patients carrying CFTR stop codon mutations. The therapeutic potential of aminoglycosides has been tested in in vitro systems, cultured cell lines, or animal models for other genetic disorders, including DMD, Hurler syndrome, nephrogenic diabetes insipidus, nephrotic cystinosis, retinitis pigmentosa, and ataxia-telangiectasia.

[0013] However, one of the major limitations of using aminoglycosides as drugs is their high toxicity to mammals, often expressed in kidney (nephrotoxicity) and ear-related (ototoxicity) diseases. The origin of this toxicity is presumed to be caused by a combination of different factors and mechanisms, such as interaction with phospholipids, inhibition of phospholipases and formation of free radicals.

[0014] Although considered selective for bacterial ribosomes, most aminoglycosides also bind to the A site of eukaryotes, but with lower affinity than the bacterial A site. Inhibition of translation in mammalian cells is also a possible reason for the high toxicity of these agents. Another factor contributing to their cytotoxicity is their binding to mitochondrial ribosomes at the 12S rRNA A site, which is very close in sequence to the bacterial A site.

[0015] Many studies have been attempted to understand and provide methods to mitigate the toxicity associated with aminoglycosides, including the use of antioxidants to reduce free radical levels, and the use of poly-L-aspartic acid and daptomycin to reduce the ability of aminoglycosides to interact with phospholipids. The role of megalin, a multiligand endocytic receptor that is particularly abundant in the proximal tubules of the kidney and the inner ear, in the uptake of aminoglycosides has recently been demonstrated. The administration of agonists that antagonize the binding of aminoglycosides to megalin also results in a reduction in aminoglycoside uptake and toxicity. In addition, changes in the administration schedule and / or the mode of administration of aminoglycosides have been studied as a means of reducing toxicity.

[0016] Despite extensive efforts to reduce aminoglycoside toxicity, few results have been matured into standard clinical practice and procedures for aminoglycoside administration to suppress stop codon mutations, except for changes in dosing regimens. For example, the use of sub-toxic doses of gentamicin in clinical trials may result in reduced readout efficiency in in vivo experiments compared to in vitro systems. (also known as G-418 sulfate or simply G-418, as shown below) showed the best termination inhibition activity in an in vitro translation-transcription system; however, aminoglycoside Its use as a therapeutic agent is unlikely because it is lethal even at very low concentrations. For example, the LD of G-418 to human fibroblasts 50 is 0.04 mg / ml. In comparison, the LD of gentamicin, neomycin and kanamycin 50 2.5-5.0mg / ml.

[0017] Because G-418 is highly toxic even at very low concentrations, gentamicin is currently the only aminoglycoside that has been tested in multiple animal models and clinical trials, while some studies have shown that amikacin and paromomycin can replace gentamicin for stop codon mutation suppression therapy.

[0018]

[0019] To date, almost all inhibition experiments have been performed using clinically available aminoglycosides, however, only a limited number of aminoglycosides, including gentamicin, amikacin, and tobramycin, are clinically used as antibiotics for internal administration in humans. Among them, tobramycin does not have stop codon mutation inhibition activity, and gentamicin is the only aminoglycoside tested for stop codon mutation inhibition activity in animal models and clinical trials. Recently, a group of neomycin derivatives have been shown to promote the readout of SMN protein in fibroblasts from patients with spinal muscular atrophy (SPA); however, these compounds were originally designed as antibiotics, and no conclusions were drawn to further improve the readout activity of these derivatives.

[0020] WO 2007 / 113841 and WO 2012 / 066546 disclose a variety of paromomycin-derived aminoglycosides designed to exhibit high readout activity of premature stop codon mutations while exerting low cytotoxicity and low antibacterial activity in mammalian cells, and thus can be used for the treatment of genetic diseases. The design of this class of paromomycin-derived aminoglycosides is achieved by introducing certain manipulations into the core of paromomycin, which can enhance the readout activity and reduce the toxicity and antibacterial activity. The manipulations are performed at several positions in the core of paromomycin.

[0021]

[0022] Exemplary such manipulations of the paromomycin core that have been taught in these disclosures include a hydroxyl group at the 6' position of the aminoglycoside core; introduction of one or more monosaccharide moieties or an oligosaccharide moiety at the 3', 4', 3, 4, 5, and / or 6 positions of the aminoglycoside core; introduction of a (S)-4-amino-2-hydroxybutyryl (AHB) moiety at the N1 position of the paromomycin core; replacement of the hydrogen at the 6' position with an alkyl group, such as a methyl substituent; and introduction of an alkyl group at the 5" position if a monosaccharide moiety is introduced into the paromomycin core.

[0023] Studies have shown that treatment with the exemplary compound NB84 disclosed in WO 2007 / 113841 (as shown in the figure below) for 2 weeks (D. Wang et al., Molecular Genetics and Metabolism, 105, 116-125 (2012); KM Keeling et al., PLoS ONE 8(4), e60478 (2013)) and 28 weeks (G. Gunn et al., Molec. Genet. Metabol. 111, 374-381 (2014)) restored sufficient α-L-iduronidase function through PTC inhibition, thereby reducing Idua tmlKmke Tissue GAG ​​accumulation in a mucopolysaccharidosis type IH (MPS IH) mouse model carrying a PTC homologous to the human IDUA-W402X nonsense mutation. The study also showed that 28 weeks of treatment with NB84 revealed significant moderation of the disease in multiple tissues resistant to current MPS IH therapies, including brain, heart, and bone. These data demonstrate that long-term nonsense suppression therapy using aminoglycosides featuring a modified paromomycin core can moderate the progression of genetic diseases.

[0024]

[0025] WO 2017 / 037717 and WO 2017 / 037718 disclose other classes of aminoglycosides derived from paromomycin, which are designed to exhibit high readout activity of premature stop codon mutations by introducing other treatments to the core of paromomycin, while exerting low cytotoxicity and low antibacterial activity in mammalian cells, which can enhance readout activity and reduce toxicity and antibacterial activity. Exemplary such treatments of the paromomycin core taught in these disclosures can be supplemented or substituted for the treatments taught in WO2007 / 113841 and WO 2012 / 066546, including further substitution of the hydroxyl group at the 6' position of the aminoglycoside core; introduction of various groups (e.g., alkyl, aryl, alkaryl, acyl, or cell permeable groups, such as guanidinium) at the N1 position of the paromomycin core; and introduction of a cell permeable group at the 5" position (if a monosaccharide is attached to the core of paromomycin).

[0026] WO 2017 / 118968 discloses other classes of aminoglycosides derived from paromomycin, which are designed to exhibit high readout activity of premature stop codon mutations by introducing other treatments to the core of paromomycin, while exerting low cytotoxicity and low antibacterial activity in mammalian cells, which can enhance readout activity and reduce toxicity and antibacterial activity. Exemplary such treatments of the paromomycin core taught in these disclosures can be supplemented or substituted for the treatments taught in WO 2007 / 113841, WO 2012 / 066546, WO 2017 / 037717 and WO 2017 / 037718, including the introduction of a hydroxyl group at the 6' position; substitution of ring I with an unsaturated ring having a double bond between the 4' and 5' positions; and the introduction of acyl groups at multiple positions.

[0027] WO 2017 / 037719 further discloses other types of paromomycin-derived aminoglycosides, which are designed to exhibit high readout activity of premature stop codon mutations by introducing other treatments into the core of paromomycin, while exerting low cytotoxicity and low antibacterial activity in mammalian cells, thereby enhancing the readout activity and reducing toxicity and antibacterial activity.

[0028] Huth et al. (J. Clin. Invest. 125, 583-92 (2015)) have hypothesized that mechanotransducer (MET) channels are present on hair cells and act as cation channels, directly involved in the entry of aminoglycosides in the cochlea (see Background Art Figure 1), and have shown that preventing / inhibiting the entry of AG through these channels can significantly reduce the ototoxic side effects of the drug. To test this hypothesis, the total positive charge of AG sisomicin was reduced by acylation of one or two amino groups of sisomicin at the same time, i.e., effectively acylation of N1 of ring II, N3 of ring III, and simultaneous acylation of N1 and N3, and the antibacterial activity and inhibition of MET channels of 9 different compounds were evaluated. It has been found that N1-mesyl modified sisomicin exhibits significantly reduced ototoxicity while maintaining antibacterial activity similar to that of the parent antibiotic sisomicin.

[0029] Other background technologies include Sabbavarapu et al., Med. Chem. Commun., 2018, 9, 503; Nudelman, I. et al., Bioorg Med Chem Lett, 2006. 16(24): p. 6310-5; Hobbie, SN et al., Nucleic Acids Res, 2007. 35(18): p. 6086-93; Kondo, J. et al., Chembiochem, 2007. 8(14): p. 1700-9; Rebibo-Sabbah, A. et al., Hum Genet, 2007. 122(3-4): p. 373-81; Azimov, R. et al., Am J Physiol. Renal Physiol, 2008.295(3):p.F633-41; Hainrichson, M. et al., Organic and Biomolecular Chemistry (OrgBiomol Chem), 2008.6(2):p.227-39; Hobbie, SN et al., Proc Natl Acad Sci USA, 2008.105(52):p.20888-93; Hobbie, SN et al., Proc Natl Acad Sci USA, 2008.105(9):p.3244-9; Nudelman, I. et al., Advanced Synthesis and Catalysis (Adv. Synth. Catal.), 2008.350:p.1682-1688; Nudelman, I. et al., J Med Chem, 2009.52(9):p.2836-45; Venkataraman, N. et al., PLoS Biol, 2009.7(4):p.e95; Brendel, C. et al., J Mol Med (Berl), 2010.89(4):p.389-98; Goldmann, T. et al., Invest Ophthalmol Vis Sci, 2010.51(12):p.6671-80; Malik, V. et al., Ther Adv Neurol Disord, 2010.3(6):p.379-89; Nudelman, I. et al., Bioorg Med Chem, 2010.18(11):p.3735-46; Warchol, ME, Curr Opin Otolaryngol Head Neck Surg, 2010.18(5): p.454-8; Lopez-Novoa, JMet al., Kidney Int., 2011.79(1): p.33-45; Rowe, SM et al., J Mol Med (Berl.), 2011.89(11): p.1149-61; Vecsler, M. et al., PLoS One, 2011.6(6): p.e20733; U.S. Patent Nos. 3,897,412, 4,024,332, 4,029,882, and 3,996,205; Greenberg et al., J. Am. Chem. Soc., 1999, 121, 6527-6541; Kotra et al., Antimicrobial Agents and Chemotherapy. chemotherapy), 2000, p.3249-3256; Haddad et al., J. Am. Chem. Soc., 2002, 124, 3229-3237; Kandasamy, J. et al. , J.Med.Chem.2012, 55, pp.10630-10643; Duscha, S. et al., MBio, 2014, 5(5), p.e01827-14; Shulman, E. et al., J Biol Chem., 2014, 289(4), pp.2318-30; Simonson et al., ChemBioChem 3, 1223-28, 2002; Shalev et al., PNAS 110, 13333-338, (2013), M. Yusopov et al., Nature 513, 517-22 (2014); Perez-Fernandez, D. et al. at. Commun. 5, 3112 (2014); Akbergenov, R. et al. Am. Soc. Microbiol. 5, 1-10 (2014); Kato, T. et al. ACS Infect. Dis. 1, 479-486 (2016); Schalev et al. Nucleic Acids Research, 43 (17), 8601-8613 (2015); Sabbavarapu et al. ACS Med. Chem. Lett. 7, 418-423 (2016); Bidou et al.RNA Biology 14, 378-388 (2017); French Patent No. 2,427,341; Japanese Patent No. 04046189; Keeling et al., PLoS ONE 8(4): e60478, 2013; and Alroy et al., Abstracts / Molecular genetics and metabolism 2018, 123(2): S18. The teachings of all of these documents are incorporated herein by reference and are as if fully set forth herein. . SUMMARY OF THE INVENTION

[0030] The present invention relates to aminoglycosides which can be beneficially used in the treatment of genetic diseases by exhibiting high premature stop codon mutation readout activity, low toxicity and low antibacterial activity in mammalian cells, and improved bioavailability and / or cell permeability. The aminoglycosides disclosed herein are characterized by a core structure based on ring I, ring II and optionally ring III of paromomycin.

[0031] According to an aspect of some embodiments of the present invention, there is provided any compound collectively represented by formula A or B in the respective embodiments as described herein.

[0032] According to one aspect of some embodiments of the present invention, there are provided compounds collectively represented by general formula I:

[0033]

[0034] in:

[0035] The dotted line indicates the stereo configuration of the 6′ position of the R configuration or the S configuration;

[0036] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl;

[0037] R2 is selected from hydrogen, substituted or unsubstituted alkyl and ORx, wherein Rx is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, or alternatively, R2 is said ORx and together with R3 forms dioxane;

[0038] R3 is selected from hydrogen, substituted or unsubstituted alkyl and ORy, wherein Ry is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, or alternatively, R3 is said ORy and together with R2 forms dioxane;

[0039] R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and

[0040] R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl,

[0041] Provided that at least one of R7-R9 is a sulfonyl group.

[0042] According to some embodiments of any of the embodiments described herein, R7 is the sulfonyl group, and the compounds are collectively represented by Formula Ia:

[0043]

[0044] in:

[0045] R1-R6, R8 and R9 are as defined in Formula I; and

[0046] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0047] According to some embodiments of any of the embodiments described herein, R' is selected from unsubstituted alkyl and unsubstituted aryl.

[0048] According to some embodiments of any of the embodiments described herein, R' is methyl.

[0049] According to some embodiments of any of the embodiments described herein, R8 and R9 are each hydrogen.

[0050] According to some embodiments of any of the embodiments described herein, R2 is ORx, and Rx is selected from hydrogen and substituted or unsubstituted alkyl.

[0051] According to some embodiments of any of the embodiments described herein, R3 is ORy, and Ry is selected from hydrogen and substituted or unsubstituted alkyl.

[0052] According to some embodiments of any of the embodiments described herein, R2 and R3 together form dioxane.

[0053] According to some embodiments of any of the embodiments described herein, there are provided compounds collectively represented by Formula Ic or Id in any corresponding embodiment as described herein.

[0054] According to one aspect of some embodiments of the present invention, there are provided compounds collectively represented by the general formula I*:

[0055]

[0056] in:

[0057] The dotted line indicates the stereo configuration of the 6′ position of the R configuration or the S configuration;

[0058] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl;

[0059] R2 is ORx, wherein Rx is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl;

[0060] R3 is ORy, wherein Ry is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl;

[0061] R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and

[0062] R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl,

[0063] And wherein the ORx and the ORy are connected to each other so that R2 and R3 together form dioxane.

[0064] According to some embodiments of any of the embodiments described herein, the dioxane is a substituted or unsubstituted 1,3-dioxane.

[0065] According to some embodiments of any of the embodiments described herein, the compounds are collectively represented by Formula I*a:

[0066]

[0067] in:

[0068] R1, R4-R6 and R7-R9 are as defined in Formula I*; and

[0069] Rw is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0070] According to some embodiments of any of the embodiments described herein, Rw is selected from substituted or unsubstituted alkyl and substituted or unsubstituted aryl.

[0071] According to some embodiments of any of the embodiments described herein, R7-R9 are each hydrogen.

[0072] According to some embodiments of any of the embodiments described herein, R8 and R9 are each hydrogen, and wherein R7 is selected from hydrogen, acyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aryl, amino substituted α-hydroxy acyl and sulfonyl.

[0073] According to some embodiments of any of the embodiments described herein, R7 is acyl.

[0074] According to some embodiments of any of the embodiments described herein, R7 is the sulfonyl group, and the compound is collectively represented by Formula I*b:

[0075]

[0076] in:

[0077] Rw, R1, R4-R6, R8 and R9 are as defined in Formula I*; and

[0078] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0079] According to some embodiments of any of the embodiments described herein, each of R4-R6 is ORz.

[0080] According to some embodiments of any of the embodiments described herein, each of R4-R6 is ORz, and in each of said R4-R6, Rz is hydrogen.

[0081] According to some embodiments of any of the embodiments described herein, at least one of R4-R6 is ORz and Rz is the monosaccharide moiety or the oligosaccharide moiety.

[0082] According to some embodiments of any of the embodiments described herein, R5 is ORz and Rz is the monosaccharide moiety.

[0083] According to some embodiments of any of the embodiments described herein, the monosaccharide moiety is represented by Formula II:

[0084]

[0085] The curves represent the locations of the attachments.

[0086] The dotted line indicates the stereo configuration of the 5″ position of the R configuration or the S configuration;

[0087] R 10 and R 11 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, and acyl;

[0088] R 12 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl; and

[0089] R 14 and R 15 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, sulfonyl and a cell-permeable group, or alternatively, R 14 and R 15 Together they form a heterocyclic ring.

[0090] According to some embodiments of any of the embodiments described herein, R5 is ORz and Rz is the monosaccharide moiety represented by formula II, and the compounds are collectively represented by formula III:

[0091]

[0092] in:

[0093] R1-R4 and R6-R9 are each as defined in Formula I or Formula Ia; and

[0094] R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in Formula II.

[0095] According to some embodiments of any of the embodiments described herein, R7 is the sulfonyl group, and the compound is collectively represented by Formula IIIa:

[0096]

[0097] in:

[0098] R1-R4, R6, R8 and R9 are as defined in Formula I or Formula Ia;

[0099] R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and

[0100] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0101] According to some embodiments of any of the embodiments described herein, R2 is ORx, and Rx is selected from hydrogen and substituted or unsubstituted alkyl.

[0102] According to some embodiments of any of the embodiments described herein, R3 is ORy, and Ry is selected from hydrogen and substituted or unsubstituted alkyl.

[0103] According to some embodiments of any of the embodiments described herein, R2 and R3 together form dioxane.

[0104] According to some embodiments of any of the embodiments described herein, R4 and R6 are each independently ORz.

[0105] According to some embodiments of any of the embodiments described herein, R4 and R6 are each ORz and Rz is hydrogen.

[0106] According to some embodiments of any of the embodiments described herein, R8 and R9 are each hydrogen.

[0107] According to some embodiments of any of the embodiments described herein, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0108] According to some embodiments of any of the embodiments described herein, R 10 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl.

[0109] According to some embodiments of any of the embodiments described herein, R 12 is a substituted or unsubstituted alkyl group.

[0110] According to some embodiments of any of the embodiments described herein, R 12It's methyl.

[0111] According to some embodiments of any of the embodiments described herein, the compound is selected from NB74-MeS; NB74-PhS; NB124-MeS; and NB124-PhS, as shown below.

[0112] According to some embodiments of any of the embodiments described herein, R5 is ORz and Rz is the monosaccharide moiety represented by formula II, and the compounds are collectively represented by formula III*:

[0113]

[0114] in:

[0115] R1-R4 and R6-R9 are each defined as in formula I* or I*a or I*b; and

[0116] R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in Formula II.

[0117] According to some embodiments of any of the embodiments described herein, the dioxane is a substituted or unsubstituted 1,3-dioxane, and the compound is collectively represented by Formula III*a:

[0118]

[0119] in:

[0120] R1, R4, R6, and R7-R9 are as defined in formula I* or I*a or I*b;

[0121] R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and

[0122] Rw is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0123] According to some embodiments of any of the embodiments described herein, Rw is selected from substituted or unsubstituted alkyl and substituted or unsubstituted aryl.

[0124] According to some embodiments of any of the embodiments described herein, R7-R9 are each hydrogen.

[0125] According to some embodiments of any of the embodiments described herein, R8 and R9 are each hydrogen, and wherein R7 is selected from hydrogen, acyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aryl, amino substituted α-hydroxy acyl and sulfonyl.

[0126] According to some embodiments of any of the embodiments described herein, R7 is acyl.

[0127] According to some embodiments of any of the embodiments described herein, R7 is the sulfonyl group, and the compound is collectively represented by Formula III*b:

[0128]

[0129] in:

[0130] Rw, R1, R4, R6, R8 and R9 are as defined in formula I*b;

[0131] R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and

[0132] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0133] According to some embodiments of any of the embodiments described herein, R4 and R6 are each independently ORz.

[0134] According to some embodiments of any of the embodiments described herein, R4 and R6 are each ORz and Rz is hydrogen.

[0135] According to some embodiments of any of the embodiments described herein, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0136] According to some embodiments of any of the embodiments described herein, R 10 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl.

[0137] According to some embodiments of any of the embodiments described herein, R 12 is a substituted or unsubstituted alkyl group, such as methyl.

[0138] According to some embodiments of any of the embodiments described herein, R1 is substituted or unsubstituted alkyl.

[0139] According to some of any of the embodiments described herein, R1 is methyl.

[0140] According to one aspect of some embodiments of the present invention, there is provided a compound collectively represented by formula IV:

[0141]

[0142] in:

[0143] Y is selected from oxygen and sulfur;

[0144] R 16 is selected from hydrogen, amine and ORq;

[0145] Rq is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, and a substituted or unsubstituted alkaryl group;

[0146] R3-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and

[0147] R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl.

[0148] According to some embodiments of any of the embodiments described herein, Y is oxygen.

[0149] According to some embodiments of any of the embodiments described herein, R 16 It's an amine.

[0150] According to some embodiments of any of the embodiments described herein, R 16 is ORq and Rq is hydrogen.

[0151] According to some embodiments of any of the embodiments described herein, R3-R6 are each independently ORz.

[0152] According to some embodiments of any of the embodiments described herein, in each of R3-R6, Rz is hydrogen.

[0153] According to some embodiments of any of the embodiments described herein, R7-R9 are each hydrogen.

[0154] According to some embodiments of any of the embodiments described herein, the compound is selected from NB160 and NB161, such as Fig.10 shown.

[0155] According to some embodiments of any of the embodiments described herein, at least one of R3-R6 is ORz, wherein Rz is a monosaccharide moiety or an oligosaccharide moiety.

[0156] According to some embodiments of any of the embodiments described herein, the Rz is a monosaccharide moiety represented by Formula II as defined in any of the corresponding embodiments.

[0157] According to some embodiments of any of the embodiments described herein, R5 is ORz, and Rz is the monosaccharide moiety represented by formula II, and the compounds are collectively represented by formula IVa:

[0158]

[0159] in:

[0160] The dotted line indicates the stereo configuration of the 5" position, which is independently R configuration or S configuration;

[0161] Y, R3, R4 and R6-R9 are each defined as Formula IV;

[0162] R 10 and R 11 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, and acyl;

[0163] R 12 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl; and

[0164] R 14 and R 15 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, sulfonyl and a cell-permeable group, or alternatively, R 14 and R 15 Together they form a heterocyclic ring.

[0165] According to some embodiments of any of the embodiments described herein, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0166] According to some embodiments of any of the embodiments described herein, R 10 , R 11 , R 14 and R 15 are each hydrogen, and wherein R 12 It is an alkyl group.

[0167] According to some embodiments of any of the embodiments described herein, the compound is selected from NB162, NB163, NB164 and NB165, such as Fig.10 shown.

[0168] According to some embodiments of the present invention, there are provided methods of preparing any of the compounds as described herein, the methods being performed according to the general description and exemplary procedures described below.

[0169] According to one aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound described herein in any embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof, and including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b), and a pharmaceutically acceptable carrier.

[0170] According to some embodiments of any of the embodiments described herein, the pharmaceutical composition is for treating a genetic disorder associated with a premature stop codon truncating mutation and / or a protein truncation phenotype.

[0171] According to some embodiments of any of the embodiments described herein, the pharmaceutical composition is packaged in packaging material and labeled in printed form in or on the packaging material for use in treating a genetic disorder associated with a premature stop codon truncating mutation and / or a protein truncation phenotype.

[0172] According to one aspect of some embodiments of the present invention, there is provided a method for treating a genetic disorder associated with a premature stop codon truncating mutation and / or a protein truncation phenotype, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as described herein in any embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof, and including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b).

[0173] According to one aspect of some embodiments of the present invention, there is provided a compound described herein in any embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof, and including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) for use in treating genetic disorders associated with premature stop codon truncating mutations and / or protein truncation phenotypes.

[0174] According to one aspect of some embodiments of the present invention, there is provided a compound described herein in any embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof, as well as compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) for the manufacture of a medicament for treating a genetic disorder associated with a premature stop codon truncating mutation and / or a protein truncation phenotype.

[0175] According to some embodiments of any of the embodiments described herein, the genetic disorder is selected from cystic fibrosis (CF), Duchenne muscular dystrophy (DMD), ataxia-telangiectasia, Hurler syndrome, hemophilia A, hemophilia B, Usher syndrome, Tay-Sachs, Becker muscular dystrophy (BMD), congenital muscular dystrophy (CMD), factor VII deficiency, familial atrial fibrillation, Hailey-Hailey disease, McArdle disease, mucopolysaccharidosis, nephropathic cystinosis, polycystic kidney disease, Rett syndrome, spinal muscular atrophy (SMA), cystinosis, severe epidermolysis bullosa, Dravet syndrome, X-linked nephrotic diabetes insipidus (XNDI), and X-linked retinitis pigmentosa, and cancer.

[0176] According to one aspect of some embodiments of the present invention, there is provided a method for increasing the expression level of a gene having a premature stop codon mutation, the method comprising translating the gene into a protein in the presence of a compound described herein in any corresponding embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiment of the compound and any combination thereof, and including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b).

[0177] According to one aspect of some embodiments of the present invention, there is provided a compound described herein in any corresponding embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiment of the compound and any combination thereof, and including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) for increasing the expression level of a gene having a premature stop codon mutation.

[0178] According to one aspect of some embodiments of the present invention, there is provided a use of a compound described herein in any corresponding embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiment of the compound and any combination thereof, as well as a compound represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) in the manufacture of a medicament for increasing the expression level of a gene having a premature stop codon mutation.

[0179] According to some of any of the embodiments described herein, the premature stop codon mutation has an RNA codon selected from the group consisting of UGA, UAG, and UAA.

[0180] According to some of any of the embodiments described herein, the protein is translated in a cytoplasmic translation system.

[0181] According to some of any of the embodiments described herein, the compound is used in a mutation inhibiting amount.

[0182] According to some of any of the embodiments described herein, the IC of the translation inhibition of the compound in the cytoplasmic translation system of a eukaryotic organism 50 Greater than the IC of translation inhibition of the compound in the ribosomal translation system 50 .

[0183] According to one aspect of some embodiments of the present invention, there is provided a compound described herein in any corresponding embodiment and any combination thereof (e.g., a compound of formula A, B, I, I*, III, III*, IV or IVa, preferably a compound of formula A, B, I, I*, III or III*, including any corresponding embodiment of the compound and any combination thereof, and including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) for use in attenuating nonsense mutation mRNA decay (NMD) and / or for treating a disease or disorder (e.g., cancer) in which attenuating NMD is beneficial.

[0184] Unless otherwise defined, all technical terms and / or scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the implementation or testing of embodiments of the present invention, exemplary methods and / or materials are described below. In the event of a conflict, the patent specification including definitions shall prevail. In addition, materials, methods, and examples are illustrative only and are not necessarily intended to be limiting.

[0185] BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0186] Some embodiments of the present invention are described herein by way of example only and with reference to the accompanying drawings. With specific reference now to the drawings in detail, it should be emphasized that the details shown are by way of example and for the purpose of illustrative discussion of embodiments of the present invention. In this regard, the description in conjunction with the drawings enables those skilled in the art to understand how to implement embodiments of the present invention.

[0187] In the attached picture:

[0188] Figure 1 (background art) shows the structure of the aminoglycoside sisomicin (left) and a schematic diagram showing the MET channel as a cation channel (A, right), as reported by Huth et al., J. Clin. Invest. 125, 583-92 (2015).

[0189] Figure 2 Shown are chemical structures of exemplary compounds having N1 substitution according to some embodiments of the present invention, also referred to herein as Set 1 and Set 2 structures.

[0190] Figure 3 Shown is a scheme of general synthetic routes for preparing exemplary compounds having N1 substitution according to some embodiments of the present invention, also referred to herein as Group 1 and Group 2 structures.

[0191] Figure 4 A scheme describing the synthesis of an exemplary receptor with a substitution at the N1 position is shown.

[0192] Figure 5 A scheme describing an exemplary synthesis of a pseudotriose aminoglycoside having a substitution at the N1 position according to an exemplary embodiment of the present invention is shown.

[0193] Figure 6 A scheme describing an exemplary synthesis of pseudo-amino disaccharides having N1-position substitution according to an exemplary embodiment of the present invention is shown.

[0194] Fig. 7A -D shows the Figure 2 Shown is a comparative graph of in vitro readout activity data for exemplary compounds having N1 substitution according to some embodiments of the present invention compared to their respective parent compounds (lacking the N1 substitution). Fig. 7A and 7B It is shown that in the R3X mutation ( Fig. 7A ) and G542X mutation ( Figure 7B ) readout of the activities of NB74, NB74-MeS, NB74-PhS and NB74-Ac. Figure 7C and 7D showed that in R3X mutation ( Figure 7C ) and G542X mutation ( Fig.7D ) readout. Mutations R3X and G542X represent nonsense mutation context constructs (UGAC and UGAG, respectively) for the genetic diseases Usher syndrome and CF, respectively.

[0195] Figure 8Dose-response curves for loss of outer hair cells are shown depending on the aminoglycoside tested; NB74-MeS (upper panel, left), NB74-Ac (upper panel, middle), NB74-PhS (upper panel, right), NB124-Ac (lower panel, left), and NB124-MeS (lower panel, right). Hair cell loss was quantified along the entire length of the cochlear explants and the concentration of hair cells that resulted in 50% loss (LC50 Coch ).

[0196] Fig. 9A -D shows hair cell loss in cochlear explants in the presence of an exemplary aminoglycoside compound according to some embodiments of the invention. Explants of the mouse organ of Corti were incubated with the drug for 72 hours and then stained for actin. Untreated control explants ( Fig. 9A ), and 15 μM NB124 ( Fig. 9B ), 150 μM NB124-Ac ( Fig. 9C ) and 15 μM NB124-MeS ( Fig.9D ) treated explants and showed that NB124-MeS and NB124-Ac displayed essentially normal morphology. Fig.9D Arrows in the images point to small areas where outer hair cells are missing.

[0197] Fig.10 Shown are chemical structures of exemplary compounds featuring carboxyl-containing (eg, carboxylate and amide) substitution at the 6′ position according to some embodiments of the present invention.

[0198] Fig.11 Shown are schemes describing exemplary synthetic routes for preparing exemplary acceptor compounds according to some embodiments of the present invention and exemplary pseudodisaccharide compounds featuring carboxyl-containing (eg, carboxylate and amide) substitution at the 6' position.

[0199] Fig.12 Shown is a scheme describing an exemplary synthesis of pseudotrisaccharide aminoglycosides featuring carboxyl-containing (eg, carboxylate and amide) substitution at the 6' position according to some embodiments of the present invention.

[0200] Fig.13 Shown are chemical structures of exemplary compounds featuring substitution at the 4' position or at the 4' and 6' positions, also referred to herein as Group 3 and Group 4 structures, according to some embodiments of the present invention.

[0201] Fig.14Shown are chemical structures of exemplary compounds featuring substitution at the 4' position or at the 4' and 6' positions, and substitution at the N1 position, also referred to herein as Group 5 and Group 6 structures, according to some embodiments of the present invention.

[0202] Fig.15 A scheme showing a general synthetic route for preparing exemplary compounds featuring substitutions at the 4' and 6' positions according to some embodiments of the present invention, also referred to herein as Group 3.

[0203] Fig.16 Shown is a scheme describing an exemplary synthetic route for preparing acceptor compounds featuring substitutions at the 4' and 6' positions according to some embodiments of the present invention; shown in the inset are structures of exemplary donor compounds according to some embodiments of the present invention.

[0204] Fig.17 A scheme showing a general synthetic route for preparing exemplary compounds featuring substitution at the 4' position according to some embodiments of the present invention, also referred to herein as Group 4, is shown.

[0205] Fig.18A -B shows a scheme describing an exemplary synthetic route for preparing acceptor compounds featuring substitution at the 4' position according to some embodiments of the present invention.

[0206] Fig.19 Schemes showing general synthetic routes for preparing exemplary compounds featuring substitution at the 4' position or at the 4' and 6' positions, and substitution at the N1 position, also referred to herein as Group 5 and Group 6 structures, according to some embodiments of the present invention.

[0207] Fig. 20 Shown is a scheme describing an exemplary synthesis of acceptor compounds featuring substitutions at the 4' and 6' positions, as well as substitutions at N1, according to some embodiments of the present invention.

[0208] Description of the Specific Embodiments of the Invention

[0209] The present invention, in some embodiments thereof, relates to aminoglycosides, and more particularly, but not exclusively, to novel aminoglycoside derivatives and their use in increasing the expression of a gene having a stop codon mutation and / or in treating genetic disorders.

[0210] Specifically, the present invention, in some embodiments thereof, relates to novel aminoglycoside compounds derived from paromomycin, exhibiting high readout activity of premature stop codon mutations, and characterized by reduced toxicity (e.g., ototoxicity) in mammalian cells. Embodiments of the present invention also relate to pharmaceutical compositions comprising these compounds, and their use in treating genetic disorders. Embodiments of the present invention also relate to methods for preparing these compounds.

[0211] The principles and operation of the present invention may be better understood with reference to the drawings and accompanying descriptions.

[0212] Before explaining in detail at least one embodiment of the present invention, it should be understood that the application of the present invention is not limited to the details as described below or illustrated by the examples. The present invention may have other embodiments or be implemented or executed in a variety of ways. Moreover, it should be understood that the wording and terminology used herein are for descriptive purposes and should not be considered as limiting.

[0213] As discussed above, the use of aminoglycosides as therapeutic agents is limited primarily due to their high toxicity. In the context of treating genetic disorders, such use is also limited by the antibacterial activity exhibited by aminoglycosides, which can also translate into toxicity.

[0214] Other limitations associated with aminoglycosides include low bioavailability, which generally requires intravenous or subcutaneous administration, and poor eukaryotic cell permeability, which generally requires high-dose administration that is associated with adverse side effects. The high aqueous solubility and polarity of aminoglycosides are hypothesized to limit their absorption through intestinal tissues and their permeability through cell membranes.

[0215] As discussed further above, several structural manipulations of the paromycin structure have resulted in synthetic aminoglycosides that have been shown to exert improved readout activity for premature stop codon mutations while exerting low toxicity in mammalian cells. WO 2007 / 113841, WO 2012 / 066546, WO 2017 / 037717, WO 2017 / 037718, WO 2017 / 037719 and WO 2017 / 118968 describing such aminoglycosides are all incorporated herein by reference and as if fully set forth herein. Two promising drug candidates disclosed in these documents are NB74 and NB124, whose structures are shown below.

[0216]

[0217] While further deciphering the structure-activity relationship of this class of aminoglycosides, in order to further improve its therapeutic effect in the context of genetic diseases, the inventors have designed a number of other modifications at different positions of the paromomycin structure, collectively represented herein by formulas A, B, I, I*, III, III*, IV and IVa. The inventors have studied the effects of these modifications on the readout activity and toxicity of the designed compounds, particularly when compared to previously disclosed modified aminoglycosides having a paromomycin core (e.g., NB74 and NB124), with the goal of discovering compounds with improved therapeutic index, i.e., exhibiting a readout activity at least as high as the previously disclosed modified aminoglycosides, but showing reduced toxicity (e.g., ototoxicity).

[0218] While simplifying the practice of the present invention, exemplary novel aminoglycoside structures were designed and successfully implemented. As demonstrated in the Examples section below, these compounds have been shown to exhibit high readout activity for disease-causing nonsense mutations and reduced toxicity.

[0219] More specifically, it has been demonstrated that Figure 2 The exemplary compounds shown have a sulfonyl substitution at the N1 position of the pseudodisaccharide and pseudotrisaccharide characterized by the paromomycin core (optionally in addition to the aforementioned modifications of the paromomycin core), at least maintaining Fig. 7A -D shows high readout activity for previously disclosed aminoglycosides (e.g., NB74 and NB124 described above), but exhibits Figure 8 and 9A -D shows significantly reduced ototoxicity.

[0220] like Fig.10 and 13 As shown, the design and practice of exemplary compounds featuring modifications at the 4' and / or 6' positions have also been demonstrated, such as Fig.14 The above is optionally combined with a sulfonyl substitution at the N1 position.

[0221] Accordingly, embodiments of the present invention relate to novel aminoglycoside (AMG) compounds (also referred to herein as "aminoglycoside derivatives" or "modified aminoglycosides"), collectively represented by Formula A, B, I, I*, III, III*, IV or IVa, methods for their preparation and their use as readout inducers of premature stop codons and / or protein truncation mutations, and thereby their use in treating genetic diseases and disorders associated with such mutations.

[0222] Compound:

[0223] As described above, the novel aminoglycoside derivatives of this embodiment are characterized by a paromomycin core, and modifications are introduced into at least one or more of its C4', C6' and N1 positions, optionally combined with other modifications, such as modifications of C6', C5, N1 and 5" as described above when the aminoglycoside is a pseudotrisaccharide.

[0224] According to some embodiments of the invention, the AMG derivatives as described herein are characterized by modification at the N1 position, and according to some of these embodiments, the amine at the N1 position is substituted with an acyl or sulfonyl group. Such AMG compounds are also referred to herein as N1-substituted compounds. Exemplary such compounds are represented herein as Group 1 and Group 2 compounds (e.g., see Figure 2 ).

[0225] According to some embodiments of the present invention, the AMG derivatives described herein have modifications at one or more of the N1 position, the N2' position, the N3 position, and optionally the N5" position in the case of a pseudotrisaccharide, and according to some of these embodiments, one or more amines at these positions are substituted with an acyl or sulfonyl group. Such AMG compounds are also referred to herein as amine-substituted compounds.

[0226] According to some embodiments of the invention, the AMG derivatives as described herein are characterized by a modification at the C4' position, and according to some of these embodiments, the AMG compounds are characterized by an alkoxy or aryloxy group at the C4' position. Such AMG compounds are also referred to herein as C4' modified compounds. Exemplary such compounds are represented herein as Group 4 and Group 6 compounds (e.g., see each Fig.13 and 14 ).

[0227] According to some embodiments of the invention, the AMG derivatives as described herein are characterized by modifications at the C4' and C6' positions, and according to some of these embodiments, C4' and C6' form part of a dioxane ring, as defined herein. Such AMG compounds are also referred to herein as C4', C6' modified compounds. Exemplary such compounds are represented herein as Group 3 and Group 5 compounds (e.g., see each Fig.13 and 14 ).

[0228] According to some embodiments of the invention, the AMG derivatives as described herein are characterized by a modification at the C6' position, and according to some of these embodiments, the AMG is characterized by a carboxyl-containing group (e.g., a carboxylate or amide as defined herein) at the C6' position. Such AMG compounds are also referred to herein as C6'-modified compounds.

[0229] According to some embodiments of the invention, the AMG derivatives as described herein are characterized by modifications at position C4', C4' and C6', or C6', as described herein, in combination with modifications at one or more of position N1, position N2', position N3, and optionally position N5" in the case of pseudotrisaccharides. Exemplary such compounds are denoted herein as Group 5 and Group 6 compounds (e.g., see Fig.14 ).

[0230] According to some of any of the embodiments described herein, the AMG derivatives as described herein further feature modification of the C6' position of the paromomamine core, when applicable, by introduction of an alkyl, cycloalkyl or aryl substituent at this position as described above.

[0231] According to some of any of the embodiments described herein, the AMG derivatives as described herein are pseudodisaccharides.

[0232] According to some of any embodiments described herein, the AMG derivatives as described herein are pseudotrisaccharides and thus feature further modification of the paromomamine core by introducing a monosaccharide moiety thereto. In some of these embodiments, AMG is further characterized by modification of the 5' position by introducing an alkyl, cycloalkyl or aryl substituent at this position as described above.

[0233] According to one aspect of some embodiments of the present invention, there is provided an AMG compound collectively represented by formula A:

[0234]

[0235] in:

[0236] The dotted line represents the stereo configuration of the 6' position for the R configuration and the S configuration (if both R1 and R2 are not hydrogen);

[0237] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl;

[0238] R2 is selected from hydrogen, substituted or unsubstituted alkyl and ORx, wherein Rx is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, or alternatively, R2 and R3 together form a dioxane ring as described herein in any corresponding embodiment;

[0239] R3 is selected from hydrogen, substituted or unsubstituted alkyl, and ORy, wherein Ry is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, or alternatively, R2 and R3 together form dioxane, as described herein in any corresponding embodiment;

[0240] R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and

[0241] R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl,

[0242] The conditions are:

[0243] (i) at least one of R7-R9 is a sulfonyl group; and / or

[0244] (ii) at least one of R7 to R9 is an acyl group,

[0245] The AMG compound is modified at one or more of the amines at the N1, N3 and N2' positions as described herein,

[0246] and / or the conditions are:

[0247] (iii) R2 and R3 together form a dioxane ring, as defined herein in any corresponding embodiment; or

[0248] (iv) R3 is ORz as defined herein, wherein Rz is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl,

[0249] Such AMG compounds may feature modifications at the C4' position, or at both the C4' and C6' positions as described herein.

[0250] According to some of any of the embodiments described herein, the AMG compounds according to this embodiment are collectively represented by Formula A as described herein, with the proviso that:

[0251] (i) at least one of R7-R9 is a sulfonyl group; and / or

[0252] (iii) R2 and R3 together form a dioxane ring as defined herein in any corresponding embodiment.

[0253] According to some of any of the embodiments described herein, the AMG compounds according to this embodiment are collectively represented by Formula A as described herein, with the proviso that:

[0254] (i) at least one of R7 to R9 is an acyl group,

[0255] And the conditions are:

[0256] (iii) R2 and R3 together form a dioxane ring, as defined herein in any corresponding embodiment; or

[0257] (iv) R3 is ORz as defined herein, wherein Rz is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl.

[0258] According to some of any of the embodiments described herein, the AMG compounds according to this embodiment are collectively represented by Formula A as described herein, with the proviso that:

[0259] (i) at least one of R7-R9 is a sulfonyl group; and / or

[0260] (ii) at least one of R7 to R9 is an acyl group,

[0261] And the conditions are:

[0262] (iv) R3 is ORz as defined herein, wherein Rz is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl.

[0263] According to some of any of the embodiments described herein, the AMG compounds according to this embodiment are collectively represented by Formula A as described herein, with the proviso that:

[0264] (ii) at least one of R7 to R9 is an acyl group,

[0265] And the conditions are:

[0266] (iv) R3 is ORz as defined herein, wherein Rz is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl.

[0267] Throughout this document, in the context of embodiments where one or more of R4-R9 is a sulfonyl group, the term "sulfonyl" describes a -S(=O)2-R' group, wherein R' in the one or more sulfonyl groups is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted alkaryl.

[0268] In some of any embodiments described herein, the sulfonyl group is an alkylsulfonyl group, such that R' is a substituted or unsubstituted alkyl group. In some of these embodiments, the alkyl group is an unsubstituted alkyl group. In some of any of these embodiments, the length of the alkyl group is 1 to 10, 1 to 8, or 1 to 6, or 1 to 4 carbon atoms. In some embodiments, the alkyl group is methyl.

[0269] In some of any of the embodiments described herein, the sulfonyl group is an arylsulfonyl group, such that R' is a substituted or unsubstituted aryl group (eg, phenyl). In some of these embodiments, the aryl group is an unsubstituted aryl group (eg, unsubstituted phenyl).

[0270] Throughout this document, in the context of embodiments involving one or more of R7-R9 being a sulfonyl group, when two or more of R7-R9 are each a sulfonyl group, the sulfonyl groups may be the same (R' in each sulfonyl group is the same) or different (R' in the two or more sulfonyl groups is different).

[0271] Throughout this document, the term "dioxane", also referred to herein as a "dioxane ring" or "dioxane moiety", describes a heteroalicyclic group or moiety as defined herein, which contains at least two oxygen atoms forming part of a ring. In the context of any embodiment involving R2 and R3 forming a dioxane, the ring preferably has at least 6 atoms and can be a 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, or higher ring. In the context of any embodiment involving R2 and R3 forming a dioxane, the dioxane is 1,3-dioxane, wherein the two oxygen atoms are separated by one carbon atom. Alternatively, the two oxygen atoms in the dioxane can be separated by 2, 3, 4, 5 or more carbon atoms.

[0272] Throughout this document, the phrase "R2 and R3 together form dioxane" means that R2 is ORx as defined herein, R3 is ORy as defined herein, and Rx and Ry are connected to each other to form dioxane. When R2 and R3 form dioxane, neither Rx nor Ry is hydrogen. When R2 and R3 form dioxane, at least two oxygen atoms of the dioxane are derived from ORx and ORy. The number of carbon atoms separating the oxygen atoms in the dioxane can be determined by Rx and / or Ry. In the case where ORx and ORy are connected together to form 1,3-dioxane, one of Rx and Ry is methyl and the other is not present.

[0273] In some of any of the embodiments involving R2 and R3 forming a dioxane, Rz and Ry form a hydrocarbon group or moiety as defined herein that links at least two oxygen atoms derived from ORx and ORy.

[0274] In this article, the term "hydrocarbon" or "hydrocarbon radical" describes an organic portion that includes a carbon atom chain mainly replaced by hydrogen atoms as its basic skeleton, also referred to herein as a backbone. Hydrocarbons can be saturated or unsaturated, contain aliphatic, alicyclic and / or aromatic moieties, and can be optionally substituted with one or more substituents (other than hydrogen). Substituted hydrocarbons can have one or more substituents, each of which can independently be, for example, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxyl, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxyl, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine, as well as any other substituents as described herein.

[0275] The hydrocarbon moiety may be optionally interrupted by one or more heteroatoms other than oxygen, including but not limited to one or more nitrogen (substituted or unsubstituted, as defined herein for -NR'-) and / or sulfur atoms.

[0276] In some of any of the embodiments described herein involving hydrocarbons, the hydrocarbon is not interrupted by any heteroatoms, nor does it contain heteroatoms in its backbone, and can be an alkylene chain, or contain alkyl, cycloalkyl, aryl, alkene and / or alkyne groups, covalently linked to each other in any order.

[0277] In some of any of the embodiments described herein, R2 and R3 form 1,3-dioxane, also referred to herein as a heterocyclic acetal.

[0278] Throughout this document, the phrase "heterocyclic acetal", also referred to herein as "cyclic acetal", describes a heterocyclic ring in which one acetal carbon and two oxygen atoms attached to the acetal carbon form part of a heteroalicyclic ring, or in other words, this phrase describes a heteroalicyclic ring comprising at least two oxygen atoms attached to each other via one carbon atom.

[0279] As used herein, the phrase "1,3-dioxane" generally describes a dioxane as defined herein, wherein at least the oxygen atoms are attached to the same carbon atom. As used herein, this term encompasses any dioxane ring.

[0280] In some of any embodiments described herein, the dioxane is a six-membered ring, and in some embodiments, it is a 6-membered 1,3-dioxane (heterocyclic acetal). In these embodiments, Rx and Ry together form a substituted or unsubstituted hydrocarbon as defined herein of one carbon atom.

[0281] Throughout this article, in the context of embodiments involving one or more of R7-R9 being acyl, the term "acyl" describes a -C(=O)-R' group, wherein R' is as described herein. In some of these embodiments, R' of the acyl group is a substituted or unsubstituted alkyl or a substituted or unsubstituted aryl. In some of these embodiments, R' is an unsubstituted alkyl, preferably a short alkyl having a length of 1-4 carbon atoms. In some of these embodiments, the acyl group is an unsubstituted aryl, such as an unsubstituted phenyl.

[0282] In some of any of the embodiments described herein, the compound is a pseudodisaccharide having Ring I and Ring II as depicted in Formula A.

[0283] In these embodiments, none of R4-R6 is ORz, wherein Rz is a monosaccharide or oligosaccharide moiety.

[0284] In some of these embodiments, one or more, or all, of R4-R6 are ORz, and Rz is not a monosaccharide or a polysaccharide.

[0285] In some of these embodiments, one or more, or all, of R4-R6 are ORz, and each of the Rz in R4-R6 is hydrogen. In these embodiments, one or more, or all, of R4-R6 are hydroxyl.

[0286] Alternatively, one or more, or all, of R4-R6 are ORz, and one or more, or all, of R4-R6 are not hydrogen.

[0287] For example, in some embodiments, one or more, or all of R4-R6 are ORz, and one or more, or all of Rz are independently substituted or unsubstituted alkyl. In these embodiments, one or more, or all of R4-R6 are alkoxy as defined herein.

[0288] For example, in some embodiments, one or more, or all of R4-R6 are ORz, and one or more, or all of Rz are independently substituted or unsubstituted aryl. In these embodiments, one or more, or all of R4-R6 are aryloxy as defined herein.

[0289] In some of these embodiments, the aryl group is unsubstituted, such that, as non-limiting examples, one or more, or all, of R4-R6 can independently be phenoxy, 1-anthryloxy, 1-naphthyloxy, 2-naphthyloxy, 2-phenanthrenyloxy, and 9-phenanthrenyloxy.

[0290] In some of these embodiments, one or more of the aryl groups in one or more of ORz is a substituted aryl group, so that, as a non-limiting example, one or more of R4-R6, or all, independently can be aryloxy, wherein aryl is 2-(N-ethylamino)phenyl, 2-(N-hexylamino)phenyl, 2-(N-methylamino)phenyl, 2,4-dimethoxyphenyl, 2-acetamidophenyl, 2-aminophenyl, 2-carboxyphenyl, 2-chlorophenyl, 2-ethoxyphenyl, 2-fluorophenyl, 2-hydroxymethylphenyl, 2-hydroxyphenyl, 2-hydroxyphenyl, 2-methoxycarbonylphenyl, 2- methoxyphenyl, 2-methylphenyl, 2-N,N-dimethylaminophenyl, 2-trifluoromethylphenyl, 3-(N,N-dibutylamino)phenyl, 3-(N,N-diethylamino)phenyl, 3,4,5-trimethoxyphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3-aminophenyl, 3-biphenyl, 3-carboxyphenyl, 3-chloro-4-methoxyphenyl, 3-chlorophenyl, 3-ethoxycarbonylphenyl, 3-ethoxyphenyl, 3-fluorophenyl, 3-hydroxymethylphenyl, 3-hydroxyphenyl, 3-isopentyloxyphenyl, 3-isobutoxyphenyl, 3- Isopropoxyphenyl, 3-methoxyphenyl, 3-methylphenyl, 3-N,N-dimethylaminophenyl, 3-tolyl, 3-trifluoromethylphenyl, 4-(benzyloxy)phenyl, 4-(isopropoxycarbonyl)phenyl, 4-(N,N-diethylamino)phenyl, 4-(N,N-dihexylamino)phenyl, 4-(N,N-diisopropylamino)phenyl, 4-(N,N-dimethylamino)phenyl, 4-(N,N-di-n-pentylamino)phenyl, 4-(n-hexyloxycarbonyl)phenyl, 4-(N-methylamino)phenyl, 4-(trifluoromethyl)phenyl, 4-aminophenyl, 4-benzyloxybenzene 4-nitrophenyl, 4-nitrophenyl, 4-propionylaminophenyl, 4-tolyl, 4-trifluoromethylphenyl, and / or 4-pentanoyloxycarbonylphenyl.

[0291] In some of these embodiments, one or more, or all, of R4-R6 are ORz, and Rz is independently substituted or unsubstituted heteroaryl. In these embodiments, one or more, or all, of R4-R6 are heteroaryloxy as defined herein.

[0292] In some embodiments, as non-limiting examples, one or more, or all of R4-R6 can independently be 2-anthryloxy, 2-furanyloxy, 2-indoleoxy, 2-naphthyloxy, 2-pyridyloxy, 2-pyrimidyloxy, 2-pyrrolyloxy, 2-quinolyloxy, 2-thienyloxy, 3-furanyloxy, 3-indoleoxy, 3-thienyloxy, 4-imidazolyloxy, 4-pyridyloxy, 4-pyrimidyloxy, 4-quinolyloxy, 5-methyl-2-thienyloxy, and 6-chloro-3-pyridyloxy.

[0293] In some of any of the embodiments described herein, R3 is aryloxy or heteroaryloxy as described herein.

[0294] In some of any of the embodiments described herein, R3 is ORy, and Ry is substituted or unsubstituted alkyl or alkenyl, such as methyl, ethyl, propyl, butyl, pentyl, propenyl, 2-hydroxyethyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, and methoxymethyl.

[0295] In some of any of the embodiments described herein, R3 is ORy and Ry is hydrogen.

[0296] In some of any of the embodiments described herein, R4 is ORz and Rz is hydrogen.

[0297] In some of any of the embodiments described herein, R6 is ORz and Rz is hydrogen.

[0298] In some of any of the embodiments described herein, one or more, or all of R4-R6 are ORz.

[0299] In some of any embodiments described herein, one or more, or all, of R4-R6 are ORz, and when one or more, or all, of the Rz moieties are not hydrogen, Rz may be the same or different for each R4-R6.

[0300] In some of these embodiments, when Rz in one or more, or all, of R4-R6 is not hydrogen, Rz can independently be, for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or heteroaryl, each optionally substituted as described herein.

[0301] In some of any embodiments described herein, when one or more of R4-R6, or all of them are ORz, Rz is independently acyl as defined herein, forming ester (carboxylate) at the corresponding position. In some of these embodiments, one or more of R4-R9 is sulfonyl as defined herein.

[0302] In some of any of the embodiments described herein, when one or more of R4-R9 is acyl, R' of the acyl is alkyl or cycloalkyl or aryl, each of which is optionally substituted with one or more amine substituents.

[0303] In some of these embodiments, R' in the acyl group is a substituted alkyl group, and in some embodiments, R' is substituted by a hydroxyl group at the alpha position relative to the carbonyl group, so that the acyl group is an alpha-hydroxy-acyl group. In some of these embodiments, at least one of R4-R9 is a sulfonyl group or R3 is ORy and Ry is not hydrogen (e.g., is an alkyl group, an aryl group, a cycloalkyl group, or an alkaryl group, each optionally substituted) or R2 and R3 together form a dioxane ring as described herein in any corresponding embodiment.

[0304] In some embodiments, the α-hydroxy-acyl group is further substituted with one or more amine groups and is an amino-substituted α-hydroxy-acyl group.

[0305] In some embodiments of acyl groups as described herein, the amine substituent can be located, for example, at one or more positions β, γ, δ, and / or ω relative to the R′ portion of the acyl group.

[0306] Exemplary amino-substituted α-hydroxy-acyl groups include, but are not limited to, the residue (S)-4-amino-2-hydroxybutyryl, also referred to herein as AHB. According to some embodiments of the present invention, an alternative to the AHB residue may be an α-hydroxy-β-aminopropionyl (AHP) residue. Other exemplary amino-substituted α-hydroxy-acyl groups include, but are not limited to, L-(-)-γ-amino-α-hydroxybutyryl, L(-)-δ-amino-α-hydroxypentanoyl, L-(-)-β-benzyloxycarbonylamino-α-hydroxypropanoyl, L-(-)-δ-benzyloxycarbonylamino-α-hydroxypentanoyl.

[0307] It should be noted herein that according to some embodiments of the present invention, other moieties involving combinations of carbonyl, hydroxyl, and amino groups and lower alkyl groups exhibiting any stereochemistry are contemplated as optional substituents replacing AHB and / or AHP, including, for example, 2-amino-3-hydroxybutyryl, 3-amino-2-hydroxypentanoyl, 5-amino-3-hydroxyhexanoyl, and the like.

[0308] In some of any of the embodiments described herein, one or more of R4-R6 is not ORz. In some of any of the embodiments described herein, one or more of R4-R6 is hydrogen.

[0309] In some of any of the embodiments described herein, R3 is hydrogen.

[0310] In some of any of the embodiments described herein, R4 is hydrogen.

[0311] In some of any of the embodiments described herein, R3 and R4 are each hydrogen.

[0312] In some of any of the embodiments described herein, one or more of R4-R6 is ORz, and Rz is independently a monosaccharide moiety or an oligosaccharide moiety as defined herein, such that the compound is a pseudotrisaccharide, pseudotetrasaccharide, pseudopentasaccharide, pseudohexasaccharide, etc.

[0313] Whenever one or more of R4-R6 is ORz and Rz is a monosaccharide moiety or an oligosaccharide moiety, and one or more of R4-R6 is not ORz wherein Rz is a monosaccharide moiety or an oligosaccharide moiety, one or more of R4-R6 that is not ORz wherein Rz is a monosaccharide moiety or an oligosaccharide moiety may be as described herein for any corresponding embodiment of R4-R6.

[0314] As used herein, the term "monosaccharide" is well known in the art and refers to a simple form of sugar consisting of a single sugar molecule that cannot be further broken down by hydrolysis. The most common examples of monosaccharides include glucose (dextrose), fructose, galactose, and ribose. Monosaccharides can be classified according to the number of carbon atoms of the carbohydrate, i.e., trisaccharides with 3 carbon atoms, such as glyceraldehyde and dihydroxyacetone; tetrasaccharides with 4 carbon atoms, such as erythrose, threose and erythrulose; pentoses with 5 carbon atoms, such as arabinose, lyxose, ribose, xylose, ribulose and xylulose; hexoses with 6 carbon atoms, such as allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose and tagatose; heptoses with 7 carbon atoms, such as mannoheptulose, sedoheptulose; octoses with 8 carbon atoms, such as 2-keto-3-deoxy-manno-octonate; nonoses with 9 carbon atoms, such as sialose; and decoses with 10 carbon atoms. Monosaccharides are the building blocks of oligosaccharides such as sucrose (common sugar) and other polysaccharides such as cellulose and starch.

[0315] As used herein, the term "oligosaccharide" refers to a compound comprising two or more monosaccharide units as defined herein linked to each other by a glycosyl bond (-O-). Preferably, the oligosaccharide comprises 2-6 monosaccharides, more preferably, the oligosaccharide comprises 2-4 monosaccharides, and most preferably, the oligosaccharide is a disaccharide moiety having two monosaccharide units.

[0316] In some of any embodiments described herein, the monosaccharide is a pentose moiety, such as represented by Formula II. Alternatively, the monosaccharide moiety is a hexose. Further alternatively, the monosaccharide moiety is not a pentose or hexose, such as a hexose moiety described in U.S. Patent No. 3,897,412.

[0317] In some of any of the embodiments described herein, the monosaccharide moiety is ribose represented by Formula II:

[0318]

[0319] The curves represent the locations of the attachments.

[0320] The dotted line indicates the stereo configuration of the 5" position of the R or S configuration:

[0321] R 10 and R 11 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, and acyl, as defined herein;

[0322] R 12 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl;

[0323] R 14 and R 15 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, sulfonyl and a cell-permeable group, or alternatively, R 14 and R 15 Together they form a heterocyclic ring.

[0324] In some of any of the embodiments described herein, R 12 For hydrogen.

[0325] In some of any of the embodiments described herein, R 12 is not hydrogen. In some of these embodiments, R 12 is alkyl, cycloalkyl or aryl, and in some embodiments, R 12 is alkyl, preferably lower alkyl, such as methyl. Alkyl, cycloalkyl or aryl may be substituted as defined herein, or unsubstituted, preferably unsubstituted.

[0326] In some of any embodiments, when one or more of R4-R6 is ORz and Rz is a monosaccharide or oligosaccharide moiety, one or more hydroxyl groups in the monosaccharide or oligosaccharide moiety / moieties are substituted with an acyl group to form an ester (carboxylate) as described herein in any corresponding embodiment.

[0327] In some of these embodiments, R 10 and R 11 One or both of the groups are acyl groups, forming an ester at the corresponding positions as described herein.

[0328] In some of any of the embodiments described herein, one of R4-R6 is ORz, and Rz is a monosaccharide moiety, such that the compound is a pseudotrisaccharide.

[0329] In some of any of the embodiments described herein for a pseudotrisaccharide, R 10 and R 11 One or more, or all, of may be acyl groups, as described herein.

[0330] In some of any of the embodiments described herein for a pseudotrisaccharide, one or more, or all, of R4-R6 are ORz, such that in one of R4-R6, Rz is a monosaccharide moiety, and in the others, Rz is as defined herein (e.g., hydrogen).

[0331] In some of any of the embodiments described herein, R5 is ORz wherein Rz is a monosaccharide moiety.

[0332] In some of these embodiments, the compound is represented by Formula B:

[0333]

[0334] wherein the variables are as described herein for Formula A, including any combination thereof.

[0335] In some of any of the embodiments described herein for Formulas A and B, R1 is hydrogen.

[0336] In some of any of the embodiments described herein for Formulas A and B, R1 is not hydrogen.

[0337] In some of any of the embodiments described herein for Formulas A and B, R1 is alkyl, and in some embodiments, R1 is lower alkyl having 1-4 carbon atoms, including but not limited to methyl, ethyl, propyl, butyl, isopropyl, and isobutyl.

[0338] In some of any of the embodiments described herein for Formulas A and B, R1 is unsubstituted alkyl.

[0339] In some of any of the embodiments described herein for Formulas A and B, R1 is methyl (eg, unsubstituted methyl).

[0340] Alternatively, in some of any of the embodiments described herein for Formulas A and B, R1 is cycloalkyl, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0341] Alternatively, in some of any of the embodiments described herein for formulas A and B, R1 is aryl, such as substituted or unsubstituted phenyl. Non-limiting examples include unsubstituted phenyl and toluene.

[0342] Alternatively, in some of any of the embodiments described herein for Formulas A and B, R1 is alkaryl, such as substituted or unsubstituted benzyl.

[0343] In some of any of the embodiments described herein for Formulas A and B, R1 is alkyl, alkenyl, or alkynyl, each substituted or unsubstituted.

[0344] In some of any embodiments described herein for Formulas A and B, R1 is or comprises an aryl group which may be substituted or unsubstituted. In some embodiments of Formulas A and B, R1 is an unsubstituted aryl group and, as non-limiting examples, may be phenyl, 1-anthryl, 1-naphthyl, 2-naphthyl, 2-phenanthrenyl or 9-phenanthrenyl.

[0345] In some embodiments of formula A and B, R1 is a substituted aryl group, and as a non-limiting example, can be 2-(N-ethylamino)phenyl, 2-(N-hexylamino)phenyl, 2-(N-methylamino)phenyl, 2,4-dimethoxyphenyl, 2-acetylaminophenyl, 2-aminophenyl, 2-carboxyphenyl, 2-chlorophenyl, 2-ethoxyphenyl, 2-fluorophenyl, 2-hydroxymethylphenyl, 2-hydroxyphenyl, 2-hydroxyphenyl, 2-methoxycarbonylphenyl, 2-methoxyphenyl, 2-methylphenyl, 2-N,N-dimethylaminophenyl, 2-trifluoromethylphenyl, phenyl, 3-(N,N-dibutylamino)phenyl, 3-(N,N-diethylamino)phenyl, 3,4,5-trimethoxyphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3-aminophenyl, 3-biphenyl, 3-carboxyphenyl, 3-chloro-4-methoxyphenyl, 3-chlorophenyl, 3-ethoxycarbonylphenyl, 3-ethoxyphenyl, 3-fluorophenyl, 3-hydroxymethylphenyl, 3-hydroxyphenyl, 3-isopentyloxyphenyl, 3-isobutoxyphenyl, 3-isopropoxyphenyl, 3-methoxyphenyl, 3-methyl phenyl, 3-N,N-dimethylaminophenyl, 3-tolyl, 3-trifluoromethylphenyl, 4-(benzyloxy)phenyl, 4-(isopropoxycarbonyl)phenyl, 4-(N,N-diethylamino)phenyl, 4-(N,N-dihexylamino)phenyl, 4-(N,N-diisopropylamino)phenyl, 4-(N,N-dimethylamino)phenyl, 4-(N,N-di-n-pentylamino)phenyl, 4-(n-hexyloxycarbonyl)phenyl, 4-(N-methylamino)phenyl, 4-(trifluoromethyl)phenyl, 4-aminophenyl, 4-benzyloxyphenyl, 4-biphenyl, 4- butoxyphenyl, 4-butyramidophenyl, 4-carboxyphenyl, 4-chlorophenyl, 4-ethoxycarbonylphenyl, 4-hexanoylaminophenyl, 4-hydroxymethylphenyl, 4-hydroxyphenyl, 4-iodophenyl, 4-isobutylphenyl, 4-isobutyramidophenyl, 4-isopropoxyphenyl, 4-isopropylphenyl, 4-methoxyphenyl, 4-methylphenyl, 4-n-hexanoylaminophenyl, 4-n-hexyloxyphenyl, 4-n-hexylphenyl, 4-nitrophenyl, 4-nitrophenyl, 4-propionamidophenyl, 4-tolyl, 4-trifluoromethylphenyl, or 4-pentanoyloxycarbonylphenyl.

[0346] In some of any of the embodiments described herein for Formulas A and B, R1 is or comprises a substituted or unsubstituted heteroaryl, and as non-limiting examples, can be 2-anthryl, 2-furyl, 2-indolyl, 2-naphthyl, 2-pyridyl, 2-pyrimidinyl, 2-pyrrolyl, 2-quinolyl, 2-thienyl, 3-furyl, 3-indolyl, 3-thienyl, 4-imidazolyl, 4-pyridyl, 4-pyrimidinyl, 4-quinolyl, 5-methyl-2-thienyl, and 6-chloro-3-pyridyl.

[0347] In some of any embodiments described herein for Formulas A and B, R1 is or comprises an amine as defined herein, and as non-limiting examples, may be -NH2, -NHCH3, -N(CH3)2, -NH-CH2-CH2-NH2, -NH-CH2-CH2-OH, and -NH-CH2-CH(OCH3)2. In some of any embodiments described herein for Formulas III or IIIa, R1 is hydroxyalkyl, such as hydroxymethyl.

[0348] In some of any of the embodiments described herein for Formulas A and B, R1 comprises a hydroxy substituent that forms a "diol-like" structure on Ring I when R2 is ORx and Rx is hydrogen.

[0349] For example, in some embodiments, R1 is configured such that the hydroxy substituent is 1-6, or 1-4, or 1-3, or 2 carbon atoms away from the hydroxy ORx group of R2.

[0350] In some of any of the embodiments described herein for Formulas A and B, R1 is hydroxy-substituted alkyl, hydroxy-substituted alkenyl, hydroxy-substituted cycloalkyl, or hydroxy-substituted aryl.

[0351] In some of any of the embodiments described herein for Formulas A and B, R1 is hydroxy-substituted alkyl, or hydroxy-substituted alkenyl, and the hydroxy substituent is a terminal substituent.

[0352] In some of any of the embodiments described herein for formulas A and B, the alkyl or alkenyl is 1-10, or 1-8, preferably 1-6, or 1-4.

[0353] In some of any embodiments of R1 in formulas A and B, R1 is hydroxyalkyl, wherein the alkyl group may be further substituted or unsubstituted.

[0354] In some of any of the embodiments described herein, R1 is hydroxymethyl.

[0355] In some of any of the embodiments described herein for Formulas A and B, the hydroxy-substituted alkyl, alkenyl, cycloalkyl or aryl group may be further substituted or unsubstituted, and may, for example, include 2 or more hydroxy groups.

[0356] In some of any of the embodiments described herein for Formulas A and B, R2 is hydrogen. In some of any of the embodiments described herein for Formulas A and B, R2 is ORx, and Rx is hydrogen.

[0357] In some of any of the embodiments described herein for Formulas A and B, R2 is ORx, and Rx is not hydrogen.

[0358] In some of any of the embodiments described herein for Formulas A and B, R2 is ORx and Rx is acyl, forming an ester at this position as described herein.

[0359] In some embodiments, R2 is ORx and Rx is alkyl, preferably selected from methyl, ethyl and propyl.

[0360] In some of any of the embodiments described herein, R2 is alkyl, and in some of these embodiments, R2 is substituted alkyl, for example, alkyl substituted with one or more amine groups (aminoalkyl).

[0361] In some of any of the embodiments described herein, R2 is substituted or unsubstituted alkyl as defined herein, or substituted or unsubstituted cycloalkyl as defined herein.

[0362] In some of any of the embodiments described herein, R2 is substituted or unsubstituted aryl as defined herein.

[0363] In some of any of the above embodiments involving R2, wherein R2 does form a dioxane ring with R3, at least one of R7-R9 is sulfonyl, and / or R3 is ORy, wherein Ry is not hydrogen, and / or at least one of R7-R9 is sulfonyl.

[0364] According to some embodiments of any of formulas A and B, the aminoglycoside structure 1 (R7), 3 (R9), 2' (R8), or 5" (R 14 and / or R 15 , if present) is modified with one or two amine substituents such that R7-R9 and R 14 and R 15 One or more of (if present) is not hydrogen.

[0365] Throughout this document, amines with substituents other than hydrogen are referred to herein as "modified amine substituents" or simply "modified amines."

[0366] According to some embodiments of the present invention, the aminoglycoside structure 1 (R7), 3 (R9), 2' (R8), or 5" (R 14 and / or R 15One or both amine substituents at the amine position (if present) are modified to include a hydrophobic moiety, such as an alkyl, cycloalkyl, alkaryl, and / or aryl group, or to include a group that is positively charged at physiological pH and that can increase the cell permeability of the compound (also interchangeably referred to herein as a "cell-permeable group" or a "cell-permealizing group"), such as a guanine or guanidine group as defined herein, or a hydrazine, hydrazide, thiohydrazide, urea and thiourea.

[0367] In some of any of the embodiments described herein, the amine substituent at position 1 (ring II) in Formula I is a modified amine as defined herein such that R7 is not hydrogen. Alternatively or additionally, one or more of R8 and R9 is not hydrogen.

[0368] In some of these embodiments, if present, R7-R9 and R 14 and R 15 One or more of are independently an alkyl group, such as a cell permeable group as described herein, or an acyl group, such as described herein in any corresponding embodiment.

[0369] If present, R7-R9 and R 14 and R 15Exemplary moieties representing one or more of include, but are not limited to, hydrogen, (R / S)-4-amino-2-hydroxybutyryl (AHB), (R / S)-3-amino-2-hydroxypropionyl (AHP), 5-aminopentanoyl, 5-hydroxypentanoyl, formyl, -C(=O)-O-methyl, -C(=O)-O-ethyl, -C(=O)-O-benzyl, -β-amino-α-hydroxypropionyl, -δ-amino-α-hydroxypentanoyl, -β-Benzyloxycarbonylamino-α-hydroxypropionyl, -δ-Benzyloxycarbonylamino-α-hydroxypentanoyl, methylsulfonyl, benzenesulfonyl, benzoyl, propyl, isopropyl, -(CH2)2NH2, -(CH2)3NH2, -CH2CH(NH2)CH3, -(CH2)4NH2, -(CH2)5NH2, -(CH2)2NH-ethyl, -(CH2)2NH(CH2)2NH2, -(CH2 )3NH(CH2)3NH2, -(CH2)3NH(CH2)4NH(CH2)3NH2, -CH(-NH2)CH2(OH), -CH(-OH)CH2(NH2), -CH (-OH)-(CH2)2(NH2), -CH(-NH2)-(CH2)2(OH), -CH(-CH2NH2)-(CH2OH), -(CH2)4NH(CH2)3NH2, -(CH2)2NH(CH2)2NH(CH2)2NH2, -(CH2)2N(CH2CH2NH2)2, -CH2-C(=O)NH2, -CH(CH3)-C(=O)NH2, -CH2-phenyl, -CH(isopropyl)-C(=O)NH2, -CH(benzyl)-C(=O)NH2, -(CH2)2OH, -(CH2)3OH, and -CH(CH2OH)2.

[0370] In some of any of the embodiments described herein, R7 is hydrogen, (R / S)-4-amino-2-hydroxybutyryl (AHB), (R / S)-3-amino-2-hydroxypropanoyl, 5-aminopentanoyl, 5-hydroxypentanoyl, formyl, -C(=O)-O-methyl, -C(=O)-O-ethyl, -C(=O)-O-benzyl, -β-amino-α-hydroxypropanoyl, -δ-amino-α-hydroxypentanoyl, -β-benzyloxy , -(CH2)2NH2, -(CH2)3NH2, -CH2CH(NH2)CH3, -(CH2)4NH2, -(CH2)5NH2, -(CH2)2NH-ethyl, -(CH2)2NH(CH2)2NH2, -(CH2)3NH H(CH2)3NH2, -(CH2)3NH(CH2)4NH(CH2)3NH2, -CH(-NH2)CH2(OH), -CH(-OH)CH2(NH2), -CH(- OH)-(CH2)2(NH2), -CH(-NH2)-(CH2)2(OH), -CH(-CH2NH2)-(CH2OH), -(CH2)4NH(CH2)3NH2,- (CH2)2NH(CH2)2NH(CH2)2NH2, -(CH2)2N(CH2CH2NH2)2, -CH2-C(=O)NH2, -CH(CH3)-C(=O)NH2, -CH2-phenyl, -CH(isopropyl)-C(=O)NH2, -CH(benzyl)-C(=O)NH2, -(CH2)2OH, -(CH2)3OH, or -CH(CH2OH)2.

[0371] In some of any of the embodiments described herein, one or both of R8 and R9 are independently hydrogen, (R / S)-4-amino-2-hydroxybutyryl (AHB), (R / S)-3-amino-2-hydroxypropionate (AHP), (R / S)-3-amino-2-hydroxypropionyl, 5-aminopentanoyl, 5-hydroxypentanoyl, formyl, -COO-methyl, -COO-ethyl, -COO-benzyl, -β-amino-α-hydroxypropionyl, - δ-amino-α-hydroxypentanoyl, -β-benzyloxycarbonylamino-α-hydroxypropionyl, -δ-benzyloxycarbonylamino-α-hydroxypentanoyl, methylsulfonyl, benzenesulfonyl, benzoyl, propyl, isopropyl, -(CH2)2NH2, -(CH2)3NH2, -CH2CH(NH2)CH3, -(CH2)4NH2, -(CH2)5NH2, -(CH2)2NH-ethyl, -(CH2)2NH(CH2)2 NH2, -(CH2)3NH(CH2)3NH2, -(CH2)3NH(CH2)4NH(CH2)3NH2, -CH(-NH2)CH2(OH), -CH(-OH)CH2(N H2), -CH(-OH)-(CH2)2(NH2), -CH(-NH2)-(CH2)2(OH), -CH(-CH2NH2)-(CH2OH), -(CH2)4NH(CH2) 3NH2, -(CH2)2NH(CH2)2NH(CH2)2NH2, -(CH2)2N(CH2CH2NH2)2, -CH2-C(=O)NH2, -CH(CH3)-C(=O)NH2, -CH2-phenyl, -CH(isopropyl)-C(=O)NH2, -CH(benzyl)-C(=O)NH2, -(CH2)2OH, -(CH2)3OH, or -CH(CH2OH)2.

[0372] In some of any of the embodiments described herein, the amino-substituted α-hydroxy acyl group is (S)-4-amino-2-hydroxybutyryl (AHB).

[0373] In some of any of the embodiments described herein, if present, R7-R9 and R 14 and R 15 One or more of is a cell permeable group as defined herein, and in some embodiments is a guanidyl group as defined herein.

[0374] In some of any of the embodiments described herein, if present, R7-R9 and R 14 and R 15 One or more of is a hydrophobic moiety, such as an alkyl group, a cycloalkyl group, an alkaryl group, and / or an aryl group.

[0375] In some of any of the embodiments described herein, if present, R4-R9 and R 14 and R 15 One or more of is an acyl group as defined herein in the corresponding embodiments, and in some of these embodiments, the acyl group may independently be an amino-substituted α-hydroxy acyl group as defined herein.

[0376] In some embodiments, wherein R4-R9, if present, and R 14 and R 15 One or more of the groups is an alkyl group, which can be, for example, a lower alkyl group of 1-4 carbon atoms, such as, but not limited to, methyl, ethyl, propyl, butyl, isopropyl, and isobutyl, each of which is optionally substituted as described herein.

[0377] In some of these embodiments, the alkyl groups are independently unsubstituted alkyl groups such as, but not limited to, ethyl, propyl, and isopropyl.

[0378] In some of these embodiments, the alkyl groups are independently substituted methyl groups, such as, but not limited to, alkaryl groups, eg, benzyl groups.

[0379] Or, if present, R4-R9 and R 14 and R 15 One or more of are independently cycloalkyl, and the cycloalkyl may be, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0380] Cross or, if present, R7-R9 and R 14 and R 15 One or more of are independently aryl, and the aryl can be, for example, substituted or unsubstituted phenyl. Non-limiting examples include unsubstituted phenyl and toluene.

[0381] In some of any of the embodiments described herein, the amine substituent at position 1 (R7, Ring II) in Formula A or B is a modified amine as described herein such that R7 is not hydrogen.

[0382] In some of these embodiments, R7 can be alkyl, cycloalkyl, alkaryl, aryl, acyl, or amino substituted α-hydroxyacyl as defined herein, for example, (S)-4-amino-2-hydroxybutyryl (AHB), or (S)-4-amino-2-hydroxypropanoyl (AHP).

[0383] In some embodiments, where R7 is alkyl, the alkyl group can be, for example, a lower alkyl group of 1-4 carbon atoms, such as, but not limited to, methyl, ethyl, propyl, butyl, isopropyl, and isobutyl, each optionally substituted as described herein.

[0384] In some of these embodiments, the alkyl groups are independently unsubstituted alkyl groups such as, but not limited to, ethyl, propyl, and isopropyl.

[0385] In some of these embodiments, the alkyl groups are independently substituted methyl groups, such as, but not limited to, alkaryl groups, eg, benzyl groups.

[0386] Alternatively, R7 is a cycloalkyl group, and the cycloalkyl group may be, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0387] Alternatively, R7 is an aryl group, and the aryl group can be, for example, a substituted or unsubstituted phenyl group. Non-limiting examples include unsubstituted phenyl and toluene.

[0388] In some of any of the embodiments described herein, R7 is alkyl, cycloalkyl, or aryl as described herein.

[0389] In some of any of the embodiments described herein, R7 is a cell permeable group as defined herein, and in some embodiments, R7 is guanidine.

[0390] In some of any embodiments of Formula B, R 14 and R 15 In some of these embodiments, R 14 and R 15 One or both of R is a cell permeable group, such as guanidine. 14 and R 15 One or both of are, for example, alkyl, cycloalkyl, or aryl as defined for any embodiment of R7.

[0391] In some of any of the embodiments described herein, if present, between R4-R9 and R 14 and R 15 When neither is sulfonyl, then R2 and R3 form a dioxane ring as described herein.

[0392] In some of any of the embodiments described herein, if present, between R4-R9 and R 14 and R 15 When both are not sulfonyl, then R3 is ORy and Ry is not hydrogen, as described herein in any corresponding embodiment.

[0393] In some of any of the embodiments described herein throughout, whenever a variable is defined as unsubstituted aryl, the unsubstituted aryl can be, for example, phenyl, 1-anthryl, 1-naphthyl, 2-naphthyl, 2-phenanthrenyl, and / or 9-phenanthrenyl.

[0394] In some of any of the embodiments described herein, whenever a variable is defined as substituted or unsubstituted heteroaryl, the heteroaryl can be, for example, 2-anthryl, 2-furanyl, 2-indolyl, 2-naphthyl, 2-pyridyl, 2-pyrimidinyl, 2-pyrrolyl, 2-quinolyl, 2-thienyl, 3-furanyl, 3-indolyl, 3-thienyl, 4-imidazolyl, 4-pyridyl, 4-pyrimidinyl, 4-quinolyl, 5-methyl-2-thienyl, and / or 6-chloro-3-pyridyl.

[0395] In some of any of the embodiments described herein, whenever a variable is defined as substituted aryl, the aryl can be, for example, 2-(N-ethylamino)phenyl, 2-(N-hexylamino)phenyl, 2-(N-methylamino)phenyl, 2,4-dimethoxyphenyl, 2-acetylaminophenyl, 2-aminophenyl, 2-carboxyphenyl, 2-chlorophenyl, 2-ethoxyphenyl, 2-fluorophenyl, 2-hydroxymethylphenyl, 2-hydroxyphenyl, 2-hydroxyphenyl, 2-methoxycarbonylphenyl, 2-methoxyphenyl, 2-methylphenyl, 2-N,N-dimethylamino phenyl, 2-trifluoromethylphenyl, 3-(N,N-dibutylamino)phenyl, 3-(N,N-diethylamino)phenyl, 3,4,5-trimethoxyphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3-aminophenyl, 3-biphenyl, 3-carboxyphenyl, 3-chloro-4-methoxyphenyl, 3-chlorophenyl, 3-ethoxycarbonylphenyl, 3-ethoxyphenyl, 3-fluorophenyl, 3-hydroxymethylphenyl, 3-hydroxyphenyl, 3-isopentyloxyphenyl, 3-isobutoxyphenyl, 3-isopropoxyphenyl, 3-methoxyphenyl, 3-methylphenyl, 3-N,N-dimethylaminophenyl, 3-tolyl, 3-trifluoromethylphenyl, 4-(benzyloxy)phenyl, 4-(isopropoxycarbonyl)phenyl, 4-(N,N-diethylamino)phenyl, 4-(N,N-dihexylamino)phenyl, 4-(N,N-diisopropylamino)phenyl, 4-(N,N-dimethylamino)phenyl, 4-(N,N-di-n-pentylamino)phenyl, 4-(n-hexyloxycarbonyl)phenyl, 4-(N-methylamino)phenyl, 4-(trifluoromethyl)phenyl, 4-aminophenyl, 4-benzyloxyphenyl, 4-biphenylyl , 4-butoxyphenyl, 4-butyrylaminophenyl, 4-carboxyphenyl, 4-chlorophenyl, 4-ethoxycarbonylphenyl, 4-hexanoylaminophenyl, 4-hydroxymethylphenyl, 4-hydroxyphenyl, 4-iodophenyl, 4-isobutylphenyl, 4-isobutyrylaminophenyl, 4-isopropoxyphenyl, 4-isopropylphenyl, 4-methoxyphenyl, 4-methylphenyl, 4-hexanoylaminophenyl, 4-n-hexyloxyphenyl, 4-n-hexylphenyl, 4-nitrophenyl, 4-nitrophenyl, 4-propionylaminophenyl, 4-tolyl, 4-trifluoromethylphenyl, and / or 4-pentanoyloxycarbonylphenyl.

[0396] Sulfonyl compounds:

[0397] According to some of any embodiments described herein, compounds represented by formula A or B are provided, wherein at least one of R7-R9 is a sulfonyl group as defined herein. Compounds according to these embodiments are also referred to herein as "sulfonyl-containing compounds".

[0398] According to some of any of the embodiments described herein, the sulfonyl-containing compound is collectively represented by Formula I:

[0399]

[0400] in:

[0401] The dotted line indicates the stereo configuration of the 6' position which is the R configuration or the S configuration as described herein;

[0402] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl, and / or as described herein in any corresponding embodiment of Formula A or B;

[0403] R2 is selected from hydrogen, substituted or unsubstituted alkyl and ORx, wherein Rx is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, and / or as described herein in any corresponding embodiment of formula A or B, or alternatively, R2 is ORx and together with R3 forms dioxane, as described herein in any corresponding embodiment;

[0404] R3 is selected from hydrogen, substituted or unsubstituted alkyl and ORy, wherein Ry is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, and / or as described herein in any corresponding embodiment of formula A or B, or alternatively, R3 is ORy and together with R2 forms dioxane;

[0405] R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and acyl, and / or as described herein in any corresponding embodiment of Formula A or B; and

[0406] R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl, and / or as described herein in any corresponding embodiment of formula A or B,

[0407] Provided that at least one of R7-R9 is a sulfonyl group.

[0408] According to some of these embodiments, R7 is the sulfonyl group, and the compounds are collectively represented by Formula Ia:

[0409]

[0410] in:

[0411] R1-R6, R8 and R9 are as defined in formula A, B or I; and

[0412] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl, as defined herein, and sulfonyl.

[0413] In some of any of the embodiments described herein, the sulfonyl group is an alkylsulfonyl group, such that R' is a substituted or unsubstituted alkyl group. In some of these embodiments, the compound is represented by Formula Ia, and R' in Formula Ia is a substituted or unsubstituted alkyl group. In some embodiments, the sulfonyl group is a methylsulfonyl group, and R' is a methyl group.

[0414] In some of any embodiments described herein, the sulfonyl group is an arylsulfonyl group, such that R' is a substituted or unsubstituted aryl group. In some of these embodiments, the compound is represented by Formula Ia, and in Formula Ia, R' is a substituted or unsubstituted aryl group (e.g., phenyl). In some embodiments, the sulfonyl group is a phenylsulfonyl group, and R' is a phenyl group (e.g., unsubstituted phenyl).

[0415] Alternatively, in any of the embodiments described herein for Formula I, R' of the sulfonyl group is an alkaryl group, such as a substituted or unsubstituted benzyl group, or a cycloalkyl group, or an alkenyl group, or an alkynyl group, or a heteroalicyclic group, or a heteroaryl group, each of which may be optionally substituted.

[0416] According to some of any of the embodiments described herein for Formula Ia, R8 and R9 are as described herein for Formula A.

[0417] According to some of any of the embodiments described herein for Formula Ia, R8 and R9 are each hydrogen.

[0418] According to some of any of the embodiments described herein for Formula I or Ia, R2 is as described herein in any of the embodiments of Formula A or B.

[0419] According to some of any of the embodiments described herein for Formula I or Ia, R2 is ORx, and Rx is as described herein in any of the corresponding embodiments of Formula A or B.

[0420] According to some of any of the embodiments described herein for Formula I or Ia, Rx is hydrogen, such that R2 is hydroxy.

[0421] According to some of any of the embodiments described herein for Formula I or Ia, Rx is substituted or unsubstituted alkyl, such that R2 is alkoxy.

[0422] According to some of any of the embodiments described herein for Formula I or Ia, Rx is substituted or unsubstituted aryl, such that R2 is aryloxy.

[0423] According to some of any of the embodiments described herein for Formula I or Ia, R3 is as described herein in any of the corresponding embodiments of Formula A or B.

[0424] According to some of any of the embodiments described herein for Formula I or Ia, R3 is ORy, and Ry is as described herein in any of the corresponding embodiments of Formula A or B.

[0425] According to some of any of the embodiments described herein for Formula I or Ia, Ry is hydrogen, such that R3 is hydroxyl.

[0426] According to some of any of the embodiments described herein for Formula I or Ia, Ry is a substituted or unsubstituted alkyl group, such that R3 is an alkoxy group.

[0427] According to some of any of the embodiments described herein for Formula I or Ia, Ry is substituted or unsubstituted aryl, such that R3 is aryloxy.

[0428] According to some of any of the embodiments described herein for Formula I or Ia, R2 and R3 together form dioxane, as described herein in any corresponding embodiment. In some embodiments, such compounds are represented by Formula I*b as described below.

[0429] According to some of any of the embodiments described herein for Formula I or Ia, R1 is not hydrogen, as described herein in any corresponding embodiment and any combination thereof, and in some of these embodiments, R1 is alkyl, such as methyl.

[0430] According to some of any of the embodiments described herein for Formula I or Ia, each of R4-R6 is independently as described herein in any of the corresponding embodiments of Formula A or B.

[0431] According to some of any of the embodiments described herein for Formula I or Ia, each of R4-R6 is independently ORz.

[0432] According to some of any of the embodiments described herein for Formula I or Ia, each of R4-R6 is ORz, and in each of R4-R6, Rz is hydrogen, such that each of R4-R6 is hydroxyl.

[0433] According to some of any of the embodiments described herein for Formula I or Ia, at least one of R4-R6 is ORz, and Rz is a monosaccharide moiety or an oligosaccharide moiety, as described herein in any corresponding embodiment.

[0434] According to some of these embodiments, at least one of R4-R6 is ORz, and Rz is a monosaccharide moiety represented by formula II, as described herein in any corresponding embodiment.

[0435] According to some of the embodiments described herein for Formula I or Ia, R5 is ORz and Rz is a monosaccharide moiety, as described herein in any corresponding embodiment. Such compounds can be collectively represented by the general formula III:

[0436]

[0437] in:

[0438] R1-R4 and R6-R9 are each as defined herein in any corresponding embodiment and any combination thereof; and

[0439] R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in Formula II in any corresponding embodiment and any combination thereof.

[0440] According to some of any of the embodiments described herein for Formula III, R7 is the sulfonyl group, and such compounds are collectively represented by Formula IIIa:

[0441]

[0442] in:

[0443] R1-R4, R6, R8 and R9 are as defined herein in any corresponding embodiment and any combination thereof;

[0444] R 10 , R 11 , R 12 , R 14 and R 15 As defined in the formula in any corresponding embodiment and any combination thereof; and

[0445] R' is as defined herein for a sulfonyl group in any corresponding embodiment and any combination thereof.

[0446] According to some of any of the embodiments described herein for Formula III or IIIa, R2 is ORx, and Rx is selected from hydrogen and substituted or unsubstituted alkyl.

[0447] According to some of any of the embodiments described herein for Formula III or IIIa, R3 is ORy, and Ry is selected from hydrogen and substituted or unsubstituted alkyl.

[0448] According to some of any of the embodiments described herein for Formula III or IIIa, R2 and R3 together form dioxane, as described herein in any corresponding embodiment. In some embodiments, such compounds are represented by Formula III*b as described below.

[0449] According to some of any of the embodiments described herein for Formula III or IIIa, R4 and R6 are each independently ORz, as described herein in any corresponding embodiment.

[0450] According to some of any of the embodiments described herein for Formula III or IIIa, R4 and R6 are each ORz and Rz is hydrogen.

[0451] According to some of any embodiments described herein for Formula III or IIIa, R8 and R9 are each hydrogen. Alternatively, R8 and R9 are each independently as described herein in any embodiment involving modified amines.

[0452] According to some of any of the embodiments described herein for Formula III or IIIa, R1 is not hydrogen, as described herein in any corresponding embodiment and any combination thereof, and in some of these embodiments, R1 is alkyl, such as methyl.

[0453] According to some of any of the embodiments described herein for Formula IIIa, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0454] According to some of any of the embodiments described herein for Formula IIIa, R 10 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl.

[0455] According to some of any of the embodiments described herein for Formula IIIa, R 12 is a substituted or unsubstituted alkyl group.

[0456] According to some of any embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R1 is not hydrogen, as described herein in any corresponding embodiment. In some of these embodiments, R1 is alkyl, such as methyl.

[0457] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R2 is ORx and Rx is hydrogen.

[0458] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R3 is ORy and Ry is hydrogen.

[0459] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R3 is ORy and Ry is alkyl.

[0460] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R4 is ORz and Rz is hydrogen.

[0461] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R6 is ORz and Rz is hydrogen.

[0462] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R8 and R9 are each hydrogen.

[0463] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0464] According to some of any of the embodiments described herein, the sulfonyl-containing compound is represented by Formula IIIa, and according to some of these embodiments, R 10 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is not hydrogen, as described herein in any corresponding embodiment. In some of these embodiments, R 12 is an alkyl group, such as methyl.

[0465] Exemplary compounds represented by Formula IIIa according to this embodiment include, but are not limited to:

[0466]

[0467]

[0468] NB74-MeS may also be referred to herein interchangeably as NB74-N1MeS or NB74-N1-MeS; NB74-PhS may also be referred to herein interchangeably as NB74-N1PhS or NB74-N1-PhS; NB124-MeS may also be referred to herein interchangeably as NB124-N1MeS or NB124-N1-MeS; and NB124-PhS may also be referred to herein interchangeably as NB124-N1PhS or NB124-N1-PhS.

[0469] Compounds containing dioxane:

[0470] According to some of any embodiments described herein, compounds represented by general formula A or B are provided, wherein R2 and R3 together form a dioxane ring as defined herein in any corresponding embodiment. Compounds according to these embodiments are also referred to herein as "dioxane-containing compounds", or "C4' and C6' modified compounds", or "C4' / C6' modified compounds".

[0471] According to some of any of the embodiments described herein, the dioxane-containing compounds are collectively represented by the general formula I*:

[0472]

[0473] or a pharmaceutically acceptable salt thereof,

[0474] in:

[0475] The dotted line represents the stereo configuration of the 6' position which is the R configuration or the S configuration (when R1 is not hydrogen);

[0476] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl, and / or as described herein in any corresponding embodiment of Formula A or B;

[0477] R2 is ORx, wherein Rx is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl;

[0478] R3 is ORy, wherein Ry is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl;

[0479] R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and acyl, and / or as described herein in any corresponding embodiment of Formula A or B; and

[0480] R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl, and / or as described herein in any corresponding embodiment of Formula A or B or Formula I or Ia,

[0481] and wherein ORx and ORy together form dioxane as described herein in any corresponding embodiment.

[0482] In some of any of the embodiments described herein for Formula I*, ORx and ORy together form a dioxane, such that Rz and Ry are linked together to form a hydrocarbon as defined herein in any corresponding embodiment in some of any of the embodiments described herein for Formula I*, linking two oxygen atoms, such that the dioxane-containing compound can be represented by Formula I**:

[0483]

[0484] wherein A is a hydrocarbon as defined herein, and all other variables are as defined herein for formula A, B or I*.

[0485] The number of carbon atoms in the main chain of the hydrocarbon determines the number of carbon atoms in the dioxane ring. For example, when the length of the hydrocarbon is 1 carbon atom (e.g., substituted or unsubstituted methylene), the dioxane ring is a 6-membered ring. When the length of the hydrocarbon is 2 carbon atoms (e.g., substituted or unsubstituted ethylene), the dioxane ring is a 7-membered ring. When the length of the hydrocarbon is 3 carbon atoms (e.g., substituted or unsubstituted propylene), the dioxane ring is an 8-membered ring, and so on.

[0486] In some of any of the embodiments described herein for Formula I*, the dioxane is substituted or unsubstituted 1,3-dioxane.

[0487] In some of any of the embodiments described herein for Formula I**, A is substituted or unsubstituted methylene, and dioxane is substituted or unsubstituted 1,3-dioxane.

[0488] Compounds wherein the dioxane is 1,3-dioxane are also referred to herein as being characterized by a heterocycle or cyclic acetal and are collectively represented by Formula I*a:

[0489]

[0490] in:

[0491] R1, R4-R6 and R7-R9 are as defined in Formula I*, A and B; and

[0492] Rw is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

[0493] In some of any of the embodiments described herein for Formula I*a, Rw is substituted or unsubstituted alkyl, such as methyl, ethyl, propyl, butyl, isopropyl, isobutyl, pentyl, and the like, each of which is optionally substituted.

[0494] In some of any of the embodiments described herein for Formula I*a, Rw is unsubstituted alkyl as defined herein, and in some of these embodiments, alkyl is methyl.

[0495] In some of any of the embodiments described herein for Formula I*a, Rw is substituted or unsubstituted aryl, for example, unsubstituted phenyl or substituted phenyl.

[0496] In some of any of the embodiments described herein for Formula I*a, Rw is unsubstituted aryl as defined herein, and in some of these embodiments, aryl is phenyl.

[0497] In some of any of the embodiments described herein for Formula I*a, Rw is substituted aryl as defined herein, and in some of these embodiments, aryl is phenyl.

[0498] When substituted, phenyl may include one or more substituents. In some embodiments, the substituted phenyl is substituted at the para position relative to the point of attachment of the 1,3-dioxane moiety. In some embodiments, the phenyl substituent may be an alkyl or alkoxy group as defined herein. In an exemplary embodiment, Rw is p-methoxyphenyl (PMP).

[0499] According to some of any of the embodiments described herein for Formula I*, I** or I*a, R1 is not hydrogen, as described herein in any corresponding embodiment and any combination thereof, and in some of these embodiments, R1 is alkyl, such as methyl.

[0500] According to some of any of the embodiments described herein for Formula I*, I** or I*a, each of R7-R9 is independently as described herein in any of the corresponding embodiments of Formula A or B.

[0501] According to some of any of the embodiments described herein for Formula I*, I** or I*a, each of R7-R9 is hydrogen.

[0502] According to some of any of the embodiments described herein for Formula I*, I** or I*a, each of R8 and R9 is hydrogen, and R7 is not hydrogen, as described herein in any of the corresponding embodiments of Formula A.

[0503] According to some of any of the embodiments described herein for Formula I*, I** or I*a, R8 and R9 are each hydrogen, and wherein R7 is selected from hydrogen, acyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aryl, amino substituted α-hydroxy acyl and sulfonyl.

[0504] According to some of any of the embodiments described herein for Formula I*, I** or I*a, R7 is acyl as described herein in any of the corresponding embodiments.

[0505] According to some of any of the embodiments described herein for Formula I*, I** or I*a, R7 is a sulfonyl group as described herein in any corresponding embodiment, such compounds can be collectively represented by Formula I*b:

[0506]

[0507] in:

[0508] Rw, R1, R4-R6, R8 and R9 are as defined in Formula I*; and

[0509] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl, as described herein in any of the corresponding embodiments of sulfonyl.

[0510] According to some of any of the embodiments described herein for Formula I*, I**, I*a or I*b, each of R4-R6 is independently as described herein in any corresponding embodiment of Formula A or B.

[0511] According to some of any of the embodiments described herein for Formula I*, I**, I*a or I*b, each of R4-R6 is independently ORz.

[0512] According to some of any of the embodiments described herein for Formula I*, I**, I*a or I*b, each of R4-R6 is ORz, and in each of said R4-R6, Rz is hydrogen, such that each of R4-R6 is hydroxyl.

[0513] According to some of any of the embodiments described herein for Formula I*, I**, I*a or I*b, at least one of R4-R6 is ORz and Rz is a monosaccharide moiety or an oligosaccharide moiety as described herein in any corresponding embodiment.

[0514] According to some of any of the embodiments described herein for Formula I*, I**, I*a or I*b, at least one of R4-R6 is ORz and Rz is a monosaccharide moiety represented by Formula II as described herein in any corresponding embodiment.

[0515] According to some of the embodiments described herein for Formula I*, I**, I*a or I*b, R5 is ORz and Rz is a monosaccharide moiety as described herein in any corresponding embodiment. Such compounds can be collectively represented by the general formula III*:

[0516]

[0517] in:

[0518] R1-R4 and R6-R9 are each defined as Formula I* or I*a or I*b in any corresponding embodiment and any combination thereof; and

[0519] R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in Formula II in any corresponding embodiment and any combination thereof.

[0520] The compound of III* can also be represented by the formula III**:

[0521]

[0522] in:

[0523] R1, R4, R6, and R4-R9 are each defined as Formula I* or I*a or I*b in any corresponding embodiment and any combination thereof;

[0524] A is a hydrocarbon as defined herein for Formula I*; and

[0525] R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in Formula II in any corresponding embodiment and any combination thereof.

[0526] According to some of any of the embodiments described herein, the dioxane is a substituted or unsubstituted 1,3-dioxane as described herein in any corresponding embodiment, and in some of these embodiments, the compound can be collectively represented by Formula III*a:

[0527]

[0528] in:

[0529] R1, R4, R6, and R7-R9 are as defined in formula I* or I*a or I*b;

[0530] R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and

[0531] Rw is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl, as described herein in any corresponding embodiment and any combination thereof.

[0532] According to some of any of the embodiments described herein for Formula III*, III*** and III*a, R7 is sulfonyl, and the compound is represented by Formula III*b:

[0533]

[0534] in:

[0535] Rw, R1, R4, R6, R8 and R9 are as defined in formula III*a;

[0536] R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and

[0537] R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl, as described herein in any corresponding embodiment and any combination thereof.

[0538] According to some of any of the embodiments described herein for Formula III*, III**, III*a or III*b, R4 and R6 are each independently ORz as described herein in any corresponding embodiment.

[0539] According to some of any of the embodiments described herein for Formula III*, III**, III*a or III*b, R4 and R6 are each independently ORz as described herein in any corresponding embodiment, R4 and R6 are each ORz and Rz is hydrogen.

[0540] According to some of any embodiments described herein for formula III*, III**, III*a or III*b, R8 and R9 are hydrogen. Alternatively, R8 and R9 are each independently as described herein in any embodiment involving modified amines.

[0541] According to some of any of the embodiments described herein for Formula III*, III**, or III*a, R7 is hydrogen, or acyl, or as described herein in any of the embodiments involving modified amines.

[0542] According to some of any of the embodiments described herein for Formula III*, III**, III*a or III*b, R1 is not hydrogen as described herein in any corresponding embodiment and any combination thereof, and in some of these embodiments, R1 is alkyl, such as methyl.

[0543] According to some of any of the embodiments described herein for Formula III*, III**, III*a or III*b, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0544] According to some of any of the embodiments described herein for Formula III*, III**, III*a or III*b, R 10 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl.

[0545] According to some of any of the embodiments described herein for Formula III*, III**, III*a or III*b, R 12 is a substituted or unsubstituted alkyl group.

[0546] According to some of any embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R1 is not hydrogen, as described herein in any corresponding embodiment. In some of these embodiments, R1 is alkyl, such as methyl.

[0547] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R7 is acyl.

[0548] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R7 is sulfonyl.

[0549] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R4 is ORz and Rz is hydrogen.

[0550] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R6 is ORz and Rz is hydrogen.

[0551] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R8 and R9 are each hydrogen.

[0552] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R 10 , R 11 , R 12 , R 14 and R 15 Each is hydrogen.

[0553] According to some of any of the embodiments described herein, the dioxane-containing compound is represented by Formula III*a, and according to some of these embodiments, R 10 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is not hydrogen, as described herein in any corresponding embodiment. In some of these embodiments, R 12 is an alkyl group, such as methyl.

[0554] Compounds containing carboxyl groups:

[0555] According to some of any of the embodiments described herein, there are provided compounds generally represented by the general formula A or B as defined herein in any corresponding embodiment, except comprising a carboxyl-containing group at the C6′ position, thus lacking the moieties represented by R1 and R2 in formulae A and B. Compounds according to these embodiments are also referred to herein as “carboxyl-containing compounds” or “C6′-modified compounds”.

[0556] Throughout this document, when used in the context of carboxyl-containing compounds, the term "carboxyl" encompasses -C(=O)-R 16 Group and -C(=S)-R 16 , where R 16 may be hydrogen, such that the carboxyl-containing group is an aldehyde or thialdehyde; or wherein R 16 may be an amine (substituted or unsubstituted) such that the carboxyl-containing group is an amide or thioamide; or R 16 It can be ORq, Rq is hydrogen, such that the carboxyl-containing group is a carboxylic acid or a thiocarboxylic acid; or Rq is alkyl, cycloalkyl, aryl, alkaryl, heteroaryl, etc. as defined herein, each optionally substituted, such that the carboxyl-containing group is a carboxylate (e.g., ester) or a thiocarboxylate (e.g., thioester).

[0557] According to some of any of the embodiments described herein, the carboxyl-containing compound is collectively represented by the general formula IV:

[0558]

[0559] Y is selected from oxygen and sulfur;

[0560] R 16 is selected from hydrogen, amine and ORq;

[0561] Rq is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, and a substituted or unsubstituted alkaryl, as defined herein in any corresponding embodiment;

[0562] R3-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and

[0563] R7-R9 are each independently selected from hydrogen, acyl, amino substituted α-hydroxy acyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl, as defined herein in any corresponding embodiment.

[0564] According to some of any of the embodiments described herein for Formula IV, Y is oxygen.

[0565] According to some of any of the embodiments described herein for Formula IV, R 16 is an amine, -NR'R", wherein R' is as described herein and R" is as described herein for R'.

[0566] According to some of any of the embodiments described herein for Formula IV, R 16 is ORq and Rq is hydrogen.

[0567] According to some of any of the embodiments described herein for Formula IV, each of R7-R9 is independently as described herein in any of the corresponding embodiments of Formula A or B.

[0568] According to some of any of the embodiments described herein for Formula IV, each of R7-R9 is hydrogen.

[0569] According to some of any of the embodiments described herein for Formula IV, each of R7-R9 is hydrogen, and R7 is not hydrogen, as described herein in any of the corresponding embodiments of Formula A.

[0570] According to some of any of the embodiments described herein for Formula IV, R8 and R9 are each hydrogen, and wherein R7 is selected from hydrogen, acyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aryl, amino substituted α-hydroxy acyl, and sulfonyl.

[0571] According to some of any of the embodiments described herein for Formula IV, R7 is acyl as described herein in any of the corresponding embodiments.

[0572] According to some of any of the embodiments described herein for Formula IV, R7 is sulfonyl as described herein in any of the corresponding embodiments.

[0573] According to some of any of the embodiments described herein for Formula IV, each of R4-R6 is independently as described herein in any of the corresponding embodiments of Formula A or B.

[0574] According to some of any of the embodiments described herein for Formula IV, each of R4-R6 is independently ORz.

[0575] According to some of any of the embodiments described herein for Formula IV, each of R4-R6 is ORz, and in each of R4-R6, Rz is hydrogen, such that each of R4-R6 is hydroxyl.

[0576] According to some of any of the embodiments described herein for Formula IV, R3 is ORy, and Ry is hydrogen.

[0577] According to some of any of the embodiments described herein for Formula IV, R3 is ORy, and Ry is as described herein in any of the corresponding embodiments of Formula A.

[0578] Exemplary compounds represented by Formula IV include NB160 and NB161 (see, e.g., Fig.10 ).

[0579] According to some of any of the embodiments described herein for Formula IV, at least one of R4-R6 is ORz, and Rz is a monosaccharide moiety or an oligosaccharide moiety, as described herein in any corresponding embodiment.

[0580] According to some of any of the embodiments described herein for Formula IV, at least one of R4-R6 is ORz, and Rz is a monosaccharide moiety represented by Formula II, as described herein in any corresponding embodiment.

[0581] According to some of any of the embodiments described herein for Formula IV, R5 is ORz and Rz is a monosaccharide moiety, as described herein in any corresponding embodiment. Such compounds can be collectively represented by the general formula IVa:

[0582]

[0583] in:

[0584] The dotted line indicates the stereo configuration of the 5" position, which is independently R configuration or S configuration;

[0585] Y, R3, R4 and R6-R9 are each defined as Formula IV;

[0586] R 10 and R 11 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, and acyl;

[0587] R 12 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl; and

[0588] R 14 and R 15 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, sulfonyl and a cell-permeable group, or alternatively, R 14 and R 15 Together they form a heterocyclic ring,

[0589] As described herein in any corresponding embodiment of Formula B.

[0590] According to some of any of the embodiments described herein for Formula IVa, R 10 , R 11 , R 12 , R 14 and R15 Each is hydrogen.

[0591] According to some of any of the embodiments described herein for Formula IVa, R 10 , R 11 , R 14 and R 15 are each hydrogen, and wherein R 12 It is an alkyl group.

[0592] Exemplary compounds represented by Formula IVa include NB162, NB163, NB164 and NB165 (see Fig.10 ).

[0593] General description:

[0594] For any of the compounds described herein and represented by Formula A, B, I, Ia, I*, I**, I*a, I*b, III, IIIa, III*, III**, III*a, III*b, IV and IVa, it should be noted that whenever optional substituents may be present at one or more carbon positions of the aminoglycoside, for example, C6', C4', C3', C2', C1', C3, C2, C1, C6, C5, and / or C1", C2", C3", C4", and C5" positions (if present), and these substituents are not specified, these substituents may each be hydrogen, or alternatively, each independently selected from alkyl, alkenyl, alkynyl, aryl, heteroaryl and cycloalkyl, each substituted or unsubstituted, or alternatively, each may be as defined herein for R7-R9.

[0595] Embodiments of the present invention also encompass compounds as described herein and represented by formula A, B, I, Ia, III and IIIa, wherein ring I is an unsaturated ring and may be referred to as "unsaturated glucosamine (ring I)-containing" compounds. Such compounds may be represented by formula Ic or Id, as shown below:

[0596]

[0597] All variables therein are individually as defined herein for the corresponding variables in Formulas A, B, I, Ia, III and IIIa.

[0598] For any of the embodiments described herein and any combination thereof, the compound can be in the form of a salt, such as a pharmaceutically acceptable salt.

[0599] As used herein, the phrase "pharmaceutically acceptable salt" refers to a charged species of a parent compound and its counter ion, which is generally used to modify the solubility characteristics of the parent compound and / or reduce any significant irritation of the parent compound to an organism without abolishing the biological activity and properties of the administered compound. Alternatively, a pharmaceutically acceptable salt of a compound as described herein may be formed during the synthesis of the compound, for example, during the isolation of the compound from a reaction mixture or during the recrystallization of the compound.

[0600] In the context of some of the present embodiments, a pharmaceutically acceptable salt of a compound described herein may optionally be an acid addition salt comprising at least one basic (e.g., amine and / or guanidine) group of the compound, wherein the basic group is in a positively charged form (e.g., wherein the basic group is protonated), combined with at least one counter ion derived from a selected base to form a pharmaceutically acceptable salt.

[0601] Thus an acid addition salt of a compound described herein may be a complex formed between one or more basic groups of the compound and one or more equivalents of an acid.

[0602] Acid addition salts can be mono- or poly-addition salts, depending on the stoichiometric ratio between the charged groups in the compound and the counterions in the salt.

[0603] As used herein, the phrase "mono-addition salt" refers to a salt wherein the stoichiometric ratio between the counterion and the charged form of the compound is 1:1, such that for every 1 molar equivalent of the compound, the addition salt contains 1 molar equivalent of the counterion.

[0604] As used herein, the phrase "poly-addition salt" refers to a salt in which the stoichiometric ratio between the counter ion and the charged form of the compound is greater than 1:1, and is, for example, 2:1, 3:1, 4:1, etc., such that for every 1 molar equivalent of the compound, the addition salt contains 2 molar equivalents or more of the counter ion.

[0605] Non-limiting examples of pharmaceutically acceptable salts are ammonium or guanidinium cations and acid addition salts thereof.

[0606] Acid addition salts can include a variety of organic and inorganic acids such as, but not limited to, hydrochloric acid to provide hydrochloric acid addition salts, hydrobromic acid to provide hydrobromic acid addition salts, acetic acid to provide acetic acid addition salts, ascorbic acid to provide ascorbic acid addition salts, benzenesulfonic acid to provide benzenesulfonate addition salts, camphorsulfonic acid to provide camphorsulfonic acid addition salts, citric acid to provide citric acid addition salts, maleic acid to provide maleic acid addition salts, malic acid to provide malic acid addition salts, methanesulfonic acid to provide methanesulfonic acid (mesylate) addition salts, naphthalenesulfonic acid to provide naphthalenesulfonic acid addition salts, oxalic acid to provide oxalic acid addition salts, phosphoric acid to provide phosphoric acid addition salts, toluenesulfonic acid to provide p-toluenesulfonic acid addition salts, succinic acid to provide succinic acid addition salts, sulfuric acid to provide sulfuric acid addition salts, tartaric acid to provide tartaric acid addition salts, and trifluoroacetic acid to provide trifluoroacetic acid addition salts. Each of these acid addition salts may be a mono-addition salt or a poly-addition salt as defined herein.

[0607] The present embodiments also encompass any enantiomer, diastereomer, prodrug, solvate, hydrate and / or pharmaceutically acceptable salt of the compounds as described herein.

[0608] As used herein, "enantiomer" refers to a stereoisomer of a compound that is superimposable with its counterpart only by complete inversion / reflection (mirror image) of each other. Enantiomers are referred to as having "chirality" because they correspond to each other like the right and left hands. Enantiomers have the same chemical and physical properties, except when present in an environment that is itself chiral (e.g., all living systems). In the context of the present embodiment, a compound may exhibit one or more chiral centers, each of which exhibits R-configuration or S-configuration and any combination, and according to a plurality of compounds of some embodiments of the present invention, any chiral center thereof may exhibit R-configuration or S-configuration.

[0609] As used herein, the term "diastereomer" refers to stereoisomers that are not enantiomers of each other. Diastereomerism occurs when two or more stereoisomers of a compound have different configurations at one or more but not all identical (related) stereocenters and are not mirror images of each other. When two diastereomers differ from each other only at one stereocenter, they are epimers. Each stereocenter (chiral center) produces two different configurations, thus producing two different stereoisomers. In the context of the present invention, embodiments of the present invention include compounds with multiple chiral centers, which appear in any combination of stereoconfigurations, i.e., any diastereomers.

[0610] According to some of any of the embodiments described herein, the stereo configuration of each of the 6' position and the 5" position (if present) is independently the R configuration or the S configuration.

[0611] According to some of any of the embodiments described herein, the stereo configuration at the 6' position is the R configuration.

[0612] According to some of any of the embodiments described herein, the stereoconfiguration at the 5" position, if present, is the S configuration.

[0613] According to some of any of the embodiments described herein, the stereo configuration at the 6' position is R configuration, and if present, the stereo configuration at the 5" position is R configuration or S configuration.

[0614] According to some of any of the embodiments described herein, the stereoconfiguration at the 6' position is the R configuration, and the stereoconfiguration at the 5" position, if present, is the S configuration.

[0615] In the structural formula given in the full text of this article, whenever a chiral carbon has a determined (defined) R-configuration or S-configuration, its chirality is represented by a triangle dotted line or a bold line, as accepted in the art, depending on the stereo configuration shown, and there is no specification. However, it should be noted that, in addition to being reflected by the triangle dotted line or the bold line shown, the present embodiment also encompasses stereo configuration. Whenever a chiral carbon has a configuration that can be an R-configuration or an S-configuration, it is represented by a rectangular dotted line and described as such. A chiral carbon atom or its racemic mixture that can adopt an R-configuration or an S-configuration can be represented herein by a simple line or a curved (wavy) line.

[0616] In the formulae given throughout this document, substituents for 6-membered rings are shown as either axial or equatorial, but each of these substituents may adopt other configurations and all such combinations are contemplated.

[0617] The term "prodrug" refers to an agent that is converted into an active compound (active parent drug) in vivo. Prodrugs can generally be used to promote the use of parent drugs. For example, prodrugs can be bioavailable by oral administration, while parent drugs cannot. Compared with parent drugs, the solubility of prodrugs in pharmaceutical compositions can also be improved. Prodrugs are also often used to achieve sustained release of active compounds in vivo. An example of a prodrug is, but is not limited to, a compound of the present invention having one or more carboxylic acid moieties, which is administered in the form of an ester ("prodrug"). This prodrug is hydrolyzed in vivo to provide a free compound (parent drug). The selected ester can affect the solubility characteristics and hydrolysis rate of the prodrug.

[0618] The term "solvate" refers to a complex with variable stoichiometry (e.g., di-, tri-, tetra-, penta-, hexa-, etc.) formed by a solute (compound of the present invention) and a solvent, wherein the solvent does not interfere with the biological activity of the solute. Suitable solvents include, for example, ethanol, acetic acid, etc.

[0619] The term "hydrate" refers to a solvate as defined above wherein the solvent is water.

[0620] According to some of any embodiments of the present invention, for formulas A, B, I, Ia, I*, I**, I*a, I*b, III, IIIa, III*, III**, III*a, and III*B, compounds known in the art that are outside the scope of the present invention are excluded, including any documents cited in the background section of this application, which are covered by these formulas.

[0621] Exemplary compounds excluded from the scope of this embodiment include, but are not limited to, gentamicin, geneticin, fortimycin, apramycin, arbekacin, dibekacin, geneticin (G-418, G418), habekacin, kanamycin, rivamycin, paromomycin, streptomycin and tobramycin.

[0622] As used herein, the term "hydroxyl" or "hydroxy" refers to an -OH group.

[0623] As used herein, the term "amine" describes a -NR'R" group, wherein each of R' and R" is independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heteroalicyclic, aryl, heteroaryl, alkaryl, alkheteroaryl, or acyl, as these terms are defined herein. Alternatively, one or both of R' and R" can be, for example, hydroxy, alkoxy, hydroxyalkyl, trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, carbonyl, C-carboxylate, O-carboxylate, N-thiocarbamate, O-thiocarbamate, urea, thiourea, N-carbamate, O-carbamate, C-amide, N-amide, guanyl, guanidine, and hydrazine.

[0624] As used herein, the term "alkyl" describes aliphatic hydrocarbons including straight and branched groups. The alkyl group may have 1-20 carbon atoms, or 1-10 carbon atoms, and may be branched or unbranched. According to some embodiments of the present invention, the alkyl group is a lower (or lower) alkyl group (i.e., methyl, ethyl, propyl, and butyl) having 1-4 carbon atoms.

[0625] When numerical ranges are stated herein, for example, "1-10" means that the group, in this case an alkyl group, can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. In some embodiments, the alkyl group is a lower alkyl group, including 1-6 or 1-4 carbon atoms.

[0626] Alkyl groups may be substituted or unsubstituted. When substituted, the substituent may be, for example, one or more of alkyl (forming a branched alkyl group), alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halogen, trihaloalkyl, hydroxy, alkoxy, and hydroxyalkyl, as these terms are defined below. Alkyl groups substituted with aryl groups are also referred to herein as "alkaryl groups," an example of which is benzyl.

[0627] When describing "alkyl", it may also be replaced by alkenyl or alkynyl. The term "alkyl" as used herein also encompasses saturated or unsaturated hydrocarbons, thus the term also encompasses alkenyl and alkynyl.

[0628] The term "alkenyl" describes an unsaturated alkyl group as defined herein having at least two carbon atoms and at least one carbon-carbon double bond, such as allyl, vinyl, 3-butenyl, 2-butenyl, 2-hexenyl, and isopropenyl. As mentioned above, alkenyl may be substituted or unsubstituted by one or more substituents.

[0629] The term "alkynyl" as defined herein is an unsaturated alkyl group having at least two carbon atoms and at least one carbon-carbon triple bond. As noted above, an alkynyl group may be substituted with one or more substituents or may be unsubstituted.

[0630] The term "cycloalkyl" refers to a full carbon monocyclic or condensed ring (i.e., a ring sharing a pair of adjacent carbon atoms), a branched or unbranched group containing 3 or more carbon atoms, wherein one or more rings do not have a completely conjugated π electron system, and may be further substituted or unsubstituted. Exemplary cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cyclododecyl. Cycloalkyl may be substituted or unsubstituted. When substituted, the substituent may be, for example, one or more of alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halogen, trihaloalkyl, hydroxyl, alkoxy, and hydroxyalkyl, as defined below by these terms.

[0631] The term "aryl" describes an all-carbon monocyclic or fused-ring polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group with a completely conjugated π electron system. The aryl group may be unsubstituted or substituted with one or more substituents. When substituted, the substituent may be, for example, one or more of an alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halogen, trihaloalkyl, hydroxyl, alkoxy, and hydroxyalkyl, as defined below for these terms.

[0632] The term "heteroaryl" describes a monocyclic or fused ring (i.e., a ring sharing a pair of adjacent atoms) having one or more atoms (e.g., nitrogen, oxygen, and sulfur) in the ring and having a completely conjugated π electron system. Examples of heteroaryl include, but are not limited to, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, and purine. Representative examples are thiadiazole, pyridine, pyrrole, oxazole, indole, purine, etc. Heteroaryl may be unsubstituted or substituted with one or more substituents. When substituted, the substituent may be, for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halogen, trihaloalkyl, hydroxy, alkoxy, and one or more of hydroxyalkyl, as defined below for these terms.

[0633] As used herein, the term "heteroalicyclic" describes a monocyclic or fused ring group having one or more atoms (e.g., nitrogen, oxygen, and sulfur) in the ring. The ring may also have one or more double bonds. However, the ring does not have a completely conjugated π electron system. Representative examples are morpholine, piperidine, piperazine, tetrahydrofuran, tetrahydropyran, etc. Heteroalicyclic may be substituted or unsubstituted. When substituted, the substituent may be, for example, one or more of an alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halogen, trihaloalkyl, hydroxy, alkoxy, and hydroxyalkyl, as these terms are defined below.

[0634] The term "halide" as used herein refers to anions of halogen atoms, namely, F-, Cl-, Br-, and I-.

[0635] The term "halo" refers to F, Cl, Br, and I atoms as substituents.

[0636] The term "alkoxide" refers to an R'-O- anion, wherein R' is as defined above.

[0637] The term "alkoxy" refers to a -OR' group, wherein R' is alkyl or cycloalkyl as defined herein.

[0638] The term "aryloxy" refers to a -OR' group, where R' is aryl as defined herein.

[0639] The term "heteroaryloxy" refers to a -OR' group where R' is heteroaryl as defined herein.

[0640] The term "thioalkoxy" refers to a -SR' group, where R' is alkyl or cycloalkyl as defined herein.

[0641] The term "thioaryloxy" refers to a -SR' group, where R' is aryl as defined herein.

[0642] The term "thiaheteroaryloxy" refers to a -SR' group where R' is heteroaryl as defined herein.

[0643] As used herein, the term "hydroxyalkyl" refers to an alkyl group, as defined herein, substituted with one or more hydroxy groups, for example, hydroxymethyl, 2-hydroxyethyl, and 4-hydroxypentyl.

[0644] The term "aminoalkyl" as used herein refers to an alkyl group, as defined herein, substituted with one or more amino groups.

[0645] The term "alkoxyalkyl" as used herein refers to an alkyl group substituted with one or more alkoxy groups, for example, methoxymethyl, 2-methoxyethyl, 4-ethoxybutyl, n-propoxyethyl, and tert-butylethyl.

[0646] The term "trihaloalkyl" refers to -CX3, wherein X is a halogen, as defined herein. An exemplary haloalkyl is CF3.

[0647] "Guandino" or "guanidine" or "guanidinyl" or "guanidyl" refers to a -RaNC(=NRd)-NRbRc group where each Ra, Rb, Rc, and Rd may each be as defined herein for R' and R".

[0648] A "guanyl" or "guanine" group refers to a RaRbNC(=NRd)- group, where Ra, Rb, and Rd are each as defined herein for R' and R".

[0649] In some of any of the embodiments described herein, the guanidine is NH—C(═NH)—NH 2 .

[0650] In some of any of the embodiments described herein, the guanosyl group is a H2N-C(=NH)- group.

[0651] Any of the amines (including modified amines), guanidine groups, and guanine groups described herein exist in their free base form, but are intended to encompass their ionized forms at physiological pH and / or in their salts, e.g., pharmaceutically acceptable salts thereof, as described herein.

[0652] Whenever alkyl, cycloalkyl, aryl, alkaryl, heteroaryl, heteroalicyclic, acyl, and any other moiety described herein is substituted, it includes one or more substituents, each of which independently can be, but is not limited to, hydroxy, alkoxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, alkaryl, alkenyl, alkynyl, sulfonate, sulfoxide, thiosulfate, sulfate, sulfite, thiosulfite, phosphonate, cyano, nitro, azo, sulfonamide, carbonyl, thiocarbonyl, C-carboxylate, O-carboxylate, N-thiocarbamate, O-thiocarbamate, oxo, thiooxo, oxime, acyl, acyl halide, azo, azide, urea, thiourea, N-carbamate, O-carbamate, C-amide, N-amide, guanosyl, guanidine, hydrazine, and hydrazide as those terms are defined herein.

[0653] The term "cyano" describes a -C≡N group.

[0654] The term "nitro" describes a -NO2 group.

[0655] The term "sulfate" describes a terminal -OS(=O)2-OR' group, as that term is defined above, or a -OS(=O)2-O- linker, as those phrases are defined above, wherein R' is as defined above.

[0656] The term "thiosulfate" describes a -OS(=S)(=O)-OR' terminal group or a -OS(=S)(=O)-O- linker, as these phrases are defined above, wherein R' is as defined above.

[0657] The term "sulfite" describes a -OS(=O)-O-R' terminal group or a -OS(=O)-O- group linker, as these phrases are defined above, wherein R' is as defined above.

[0658] The term "thiosulfite" describes a terminal -OS(=S)-O-R' group or a linking -OS(=S)-O- group, as these phrases are defined above, wherein R' is as defined above.

[0659] The term "sulfinate" describes a -S(=O)-OR' terminal group or a -S(=O)-O- group linker, as these phrases are defined above, wherein R' is as defined above.

[0660] The term "sulfoxide" or "sulfinyl" describes a -S(=O)R' terminal group or a -S(=O)- linker, as those phrases are defined above, where R' is as defined above.

[0661] The term "sulfonate" or "sulfonyl" describes a terminal -S(=O)2-R' group or a -S(=O)2- linker, as those phrases are defined above, where R' is as defined herein.

[0662] The term "S-sulfonamide" describes a -S(=O)2-NR'R" terminal group or a -S(=O)2-NR'- linking group, as these phrases are defined above, where R' and R" are as defined herein.

[0663] The term "N-sulfonamide" describes a R'S(=O)2-NRR"-terminal group or a -S(=O)2-NR'-linking group, as these phrases are defined above, where R' and R" are as defined herein.

[0664] The term "carbonyl" or "carbonate" is used herein to describe a terminal -C(=O)-R' group or a -C(=O)- linker, as those phrases are defined above, wherein R' is as defined herein.

[0665] The term "thiocarbonyl" is used herein to describe a terminal -C(=S)-R' group or a -C(=S)- linker, as those phrases are defined above, wherein R' is as defined herein.

[0666] The term "oxo" as used herein describes a (=0) group wherein the oxygen atom is attached to the atom (eg, carbon atom) at the indicated position via a double bond.

[0667] The term "thiooxo" as used herein describes a (=S) group in which the sulfur atom is attached to the atom (eg, carbon atom) at the indicated position via a double bond.

[0668] The term "oxime" describes a =N-OH terminal group or a =NO- linking group, as those phrases are defined above.

[0669] The term "acyl halide" describes a -(C=O)R"" group, where R"" is a halide as defined above.

[0670] The terms "azo" or "diazo" describe a -N=NR' terminal group or a -N=N- linking group, as those phrases are defined above, where R' is as defined above.

[0671] The term "azide" describes a -N3 terminal group.

[0672] As used herein, the term "carboxylate" encompasses both C-carboxylates and O-carboxylates.

[0673] The term "C-carboxylate" describes a -C(=O)-OR' terminal group or a -C(=O)-O- linker, as these phrases are defined above, wherein R' is as defined herein.

[0674] The term "O-carboxylate" describes a -OC(=O)R' terminal group or a -OC(=O)- linker, as those phrases are defined above, wherein R' is as defined herein.

[0675] Carboxylates can be linear or cyclic. When cyclic, R' and the carbon atom are linked together to form a ring in the C-carboxylates, which is also referred to as a lactone. Alternatively, Ra and O are linked together to form a ring in the O-carboxylates. Cyclic carboxylates can be used as a linking group, for example, when one atom in the formed ring is linked to another group.

[0676] As used herein, the term "thiocarboxylate" encompasses both C-thiocarboxylates and O-thiocarboxylates.

[0677] The term "C-thiocarboxylate" describes a -C(=S)-OR' terminal group or a -C(=S)-O- linker, as those phrases are defined above, wherein R' is as defined herein.

[0678] The term "O-thiocarboxylate" describes a terminal -OC(=S)R' group or a -OC(=S)- linker, as those phrases are defined above, wherein R' is as defined herein.

[0679] Thiocarboxylates can be straight chain or cyclic. When cyclic, R' and carbon atom are linked together to form a ring in C-thiocarboxylates, which is also referred to as thiolactone. Alternatively, R' and O are linked together to form a ring in O-thiocarboxylates. Cyclic thiocarboxylates can be used as a linking group, for example, when an atom in the formed ring is linked to another group.

[0680] As used herein, the term "carbamate" encompasses both N-carbamates and O-carbamates.

[0681] The term "N-carbamate" describes an R"OC(=O)-NR'-terminal group or a -OC(=O)-NR'-linking group, as these phrases are defined above, where R' and R" are as defined herein.

[0682] The term "O-carbamate" describes a -OC(=O)-NR'R" terminal group or a -OC(=O)-NR'- linking group, as these phrases are defined above, wherein R' and R" are as defined herein.

[0683] Carbamates can be linear or cyclic. When cyclic, R' and the carbon atom are linked together to form a ring in an O-carbamate. Alternatively, R' and O are linked together to form a ring in an N-carbamate. Cyclic carbamates can be used as linking groups, for example, when one atom in the formed ring is linked to another group.

[0684] As used herein, the term "carbamate" encompasses both N-carbamates and O-carbamates.

[0685] As used herein, the term "thiocarbamate" encompasses both N-thiocarbamates and O-thiocarbamates.

[0686] The term "O-thiocarbamate" describes a -OC(=S)-NR'R" terminal group or a -OC(=S)-NR'- linking group, as those phrases are defined above, wherein R' and R" are as defined herein.

[0687] The term "N-thiocarbamate" describes an R"OC(=S)NR'-terminal group or a -OC(=S)NR'-linking group, as these phrases are defined above, where R' and R" are as defined herein.

[0688] The thiocarbamates may be linear or cyclic as described herein for carbamates.

[0689] As used herein, the term "dithiocarbamate" encompasses both S-dithiocarbamates and N-dithiocarbamates.

[0690] The term "S-dithiocarbamate" describes a -SC(=S)-NR'R" terminal group or a -SC(=S)NR'- linking group, as those phrases are defined above, where R' and R" are as defined herein.

[0691] The term "N-dithiocarbamate" describes a R"SC(=S)NR'-terminal group or a -SC(=S)NR'-linking group, as these phrases are defined above, where R' and R" are as defined herein.

[0692] The term "urea", also referred to herein as "ureido", describes a -NR'C(=O)-NR"R"' terminal group or a -NR'C(=O)-NR"- linking group, as these phrases are defined above, wherein R' and R" are as defined herein and R"' is as defined herein for R' and R".

[0693] The term "thiourea", also referred to herein as "thioureido", describes a -NR'-C(=S)-NR"R"' terminal group or a -NR'-C(=S)-NR"- linking group, wherein R', R", and R'" are as defined herein.

[0694] As used herein, the term "amide" encompasses both C-amides and N-amides.

[0695] The term "C-amide" describes a -C(=O)-NR'R" terminal group or a -C(=O)-NR'- linker, as these phrases are defined above, where R' and R" are as defined herein.

[0696] The term "N-amide" describes an R'C(=O)-NR"-terminal group or an R'C(=O)-N-linking group, as these phrases are defined above, wherein R' and R" are as defined herein.

[0697] The term "hydrazine" describes a -NR'-NR"R"' terminal group or a -NR'-NR"- linking group, as these phrases are defined above, where R', R", and R'" are as defined herein.

[0698] The term "hydrazide" as used herein describes a -C(=O)-NR'-NR"R"' terminal group or a -C(=O)-NR'-NR"- linker, as those phrases are defined above, where R', R", and R'" are as defined herein.

[0699] The term "thiohydrazide" as used herein describes a -C(=S)-NR'-NR"R"' terminal group or a -C(=S)-NR'-NR"- linker, as those phrases are defined above, wherein R', R", and R'" are as defined herein.

[0700] method:

[0701] Further in accordance with embodiments of the present invention, there are provided methods of preparing the compounds as described herein.

[0702] These methods are generally accomplished by providing a paromomamine derivative and introducing the desired modifications thereto to obtain a pseudodisaccharide compound as described herein.

[0703] The methods for preparing pseudotrisaccharide compounds as described herein are generally accomplished by designing appropriate acceptor aminoglycoside molecules and corresponding donor molecules, as is known in the aminoglycoside art.

[0704] Generally, the synthesis of pseudotrisaccharide compounds according to some embodiments of the present invention is accomplished using appropriate acceptor and donor molecules and reaction conditions that allow protected derivatives of the donor and / or acceptor to react and remove protecting groups to obtain the desired pseudotrisaccharide.

[0705] The term "acceptor" is used herein to describe a scaffold structure derived from paromomamine that has an available (unprotected) hydroxyl group at position C3', C4', C6 or C5, preferably C5, which is reactive during the glycosylation reaction and can accept a sugar group.

[0706] The term "donor" is used herein to describe the sugar group that reacts with the acceptor to form the final pseudotrisaccharide compound.

[0707] As used herein, the term "glycosyl" refers to a chemical group obtained by removing a hydroxyl group from the hemiacetal functional group of a monosaccharide.

[0708] According to some embodiments of the present invention, donors and acceptors are designed to form desired compounds. Some embodiments of this aspect of the present invention are described below, showing exemplary methods for preparing exemplary subsets of compounds as described herein. The following examples and accompanying drawings show more detailed methods for preparing exemplary compounds according to some embodiments of the present invention.

[0709] The synthesis of pseudotrisaccharide compounds according to some embodiments of the present invention generally comprises (i) preparing an acceptor compound by selectively protecting one or more hydroxyl groups and amine groups present at selected positions on the paromycin scaffold, leaving the selected position (e.g., C5) unprotected and thus capable of freely accepting a donor (sugar) compound as defined herein; (ii) preparing a donor compound by selectively protecting one or more hydroxyl groups and amine groups present at selected positions on the sugar group, leaving one position unprotected and thus capable of freely coupling with an acceptor compound as defined herein; (iii) subjecting the donor and the acceptor to a coupling reaction; and (iii) removing the protecting group to obtain the desired compound.

[0710] As used herein, the phrase "protected group" refers to a group that is substituted or modified to block its functionality and protect it from reacting with other groups under reaction conditions (e.g., coupling reactions as described herein). The protected group is regenerated by removing the substituent or by re-modification.

[0711] When "amino-protected group" or "hydroxyl-protected group" is used, it means that the protecting group is attached or used to modify the corresponding group to generate the protected group.

[0712] As used herein, the phrase "protecting group" refers to a substituent or modification that is typically used to block or protect a particular functionality while reacting other functional groups on the compound. Protecting groups are selected to release a substituent or be remodified to generate the desired unprotected group.

[0713] For example, an "amino-protecting group" or "amine-protecting group" is a substituent attached to an amino group, or a modification of an amino group, which blocks or protects the amino functional group in a compound and prevents it from participating in chemical reactions. The amino-protecting group is removed by removal of the substituent or by modification to regenerate an amine group.

[0714] Suitable amino protecting groups include azide (azido), N-phthalimidyl, N-acetyl, N-trifluoroacetyl, N-tert-butyloxycarbonyl (BOC), N-benzyloxycarbonyl (CBz) and N-9-fluorenylmethoxycarbonyl (Fmoc).

[0715] "Hydroxy-protecting group" or "hydroxyl-protecting group" refers to a substituent or modification of a hydroxy group that blocks or protects the hydroxyl functionality and prevents the hydroxyl functionality from participating in chemical reactions. The hydroxyl-protecting group is removed by removal of the substituent or by the modification to regenerate the hydroxyl group.

[0716] Suitable hydroxy protecting groups include isopropylidene ketal and cyclohexanone dimethyl ketal (forming 1,3-dioxane with two adjacent hydroxyl groups), 4-methoxy-1-methylbenzene (forming 1,3-dioxane with two adjacent hydroxyl groups), O-acetyl, O-chloroacetyl, O-benzoyl, and O-silyl (e.g., O-trimethylsilyl; O-TMS).

[0717] For a general description of protecting groups and their use, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0718] According to some embodiments, the amino protected group includes azide (N3-) and / or N-phthalimido, and the hydroxyl protecting group includes O-acetyl (AcO-), O-benzoyl (BzO-), O-TMS (TMSO-) and / or O-chloroacetyl.

[0719] It should be noted herein that "protected group", when used, refers to a residue in which one reactive functional group on a compound is protected or more than one reactive functional group is protected simultaneously, such as in the case of two adjacent functional groups, for example, in the case of two hydroxyl groups that can be protected at once via isopropylidene ketal.

[0720] In some embodiments, the donor compound is a protected monosaccharide represented by the general formula II*, as defined herein, having a leaving group at its 1" position, represented by L, and optionally having a substituent R at the 5" position. 12 :

[0721]

[0722] in:

[0723] L is a leaving group;

[0724] OT is the donor protected hydroxyl group;

[0725] R 12 As defined herein for Formula Ib (the 5" configuration shown in Formula III is a non-limiting example);

[0726] as well as

[0727] D is NR as defined in formula III, IIIa, III*, III*a, ITI*b, ITI** and IVa 14 R 15 The protected or unprotected form of formula III, IIIa, III*, III*a, III*b, III** and IVa wherein R 14 and R 15 When both are hydrogen, D is a donor protected amine group.

[0728] As used herein, phrase "leaving group" describes an unstable atom, group, or chemical moiety that is easily separated from an organic molecule during a chemical reaction, and separation is usually promoted by the relative stability of the leaving atom, group, or residue thereon. Usually, any group as the conjugate base of a strong acid can serve as a leaving group. Representative examples of suitable leaving groups according to some of the present embodiments include but are not limited to trichloroacetimido esters, acetates, tosylates, trifluoromethanesulfonates, sulfonates, azides, halides, hydroxyls, thiohydroxyls, alkoxyls, cyanates, thiocyanates, nitro, and cyano groups.

[0729] According to some embodiments of the invention, each donor hydroxyl protecting group is O-benzoyl, although other protecting groups are contemplated, and R 15 or R 17 The donor amino protecting group in is an azide group.

[0730] It should be noted that when R 14 and R 15 When one of R6 and R7 is not hydrogen, it may be protected or unprotected. Generally, when one of R6 and R7 is guanine or guanidine, a protecting group suitable for the reaction conditions (e.g., a coupling reaction with a receptor) may be used. Optionally, guanine or guanidine is unprotected. When R 14 and R 15 When one of them is alkyl, aryl or cycloalkyl, usually D in formula II* is NR 14 R 15 unprotected form.

[0731] According to some embodiments of the present invention, the structure of the donor compound sets the absolute structure of ring III in the resulting compound, i.e., the stereo configuration of the 5" position and R in formula III, IIIa, III*, III*a, III*b, III** and IVa. 14 , R 15 and R 12 Type.

[0732] An exemplary receptor molecule suitable for preparing compounds of Formula IIIa as described herein is represented by Formula V:

[0733]

[0734] in:

[0735] The dashed line indicates the S-configuration or R-configuration at the 6′ position;

[0736] OP is the protected hydroxyl group of the acceptor;

[0737] AP is the protected amine group of the receptor;

[0738] R1 is as defined herein for Formula I or Ia;

[0739] A is an acceptor protected hydroxyl group (OP); or, depending on the chemical nature of these groups and the reaction conditions, may be one of the other groups that define OR2, protected or unprotected; and

[0740] B is a protected or unprotected form of a group defining R7.

[0741] According to some embodiments of the present invention, the acceptor hydroxyl protecting group is an O-acetyl group.

[0742] According to some embodiments of the invention, the acceptor hydroxyl protecting group is O-TMS.

[0743] Other hydroxy protecting groups are also contemplated.

[0744] According to some embodiments of the invention, the acceptor amino protecting group is an azido group, although other protecting groups are contemplated.

[0745] The receptor hydroxyl-protected groups and the receptor amino-protected groups at multiple positions of the receptor may be the same or different at each position.

[0746] In some embodiments, the acceptor is prepared by generating moiety B prior to reaction with the donor.

[0747] According to some embodiments of the invention, the structure of the acceptor compound sets the absolute structure of Ring I and Ring II in the resulting compound.

[0748] The Examples section below and the Figure 3-6 and 15-20 show exemplary receptors suitable for preparing compounds of Formula III or IIIa.

[0749] In some embodiments, the synthesis of the pseudodisaccharide compound of formula Ia according to some embodiments of the present invention is accomplished using an amino protected compound of formula VI:

[0750]

[0751] in:

[0752] The dashed line indicates the S-configuration or R-configuration at the 6′ position;

[0753] AP is the protected amine group of the receptor;

[0754] R1 is as defined herein for Formula Ia;

[0755] A is an acceptor protected hydroxyl group (OP) as described herein; or it may be one of the other groups defining OR2, protected or unprotected, depending on the chemical nature of these groups and the reaction conditions.

[0756] The compound of formula IV is converted to the group defining R7 in formula Ia using methods known in the art.

[0757] The Examples section and Figures below show exemplary amino-protected compounds suitable for preparing compounds of Formula I or Ia (see, e.g. Figure 3-5 ).

[0758] An exemplary receptor molecule suitable for preparing compounds of Formula III*a as described herein is represented by Formula VII:

[0759]

[0760]

[0761] in:

[0762] The dashed line indicates the S-configuration or R-configuration at the 6′ position;

[0763] OP is the protected hydroxyl group of the acceptor;

[0764] AP is the protected amine group of the receptor;

[0765] R1 is as defined herein;

[0766] If R7 in Formula Ia is hydrogen, then B is an acceptor protected amine group; or may be a protected or unprotected form of a group defining R7; and

[0767] K is a protected or unprotected form of a group defining Rw.

[0768] According to some embodiments of the present invention, the acceptor hydroxyl protecting group is an O-acetyl group.

[0769] According to some embodiments of the invention, the acceptor hydroxyl protecting group is O-TMS.

[0770] Other hydroxy protecting groups are also contemplated.

[0771] According to some embodiments of the invention, the acceptor amino protecting group is an azido group, although other protecting groups are also contemplated.

[0772] The receptor hydroxyl-protected groups and the receptor amino-protected groups at multiple positions of the receptor may be the same or different at each position.

[0773] In some embodiments, the acceptor is prepared by generating moiety B prior to reaction with the donor, if applicable.

[0774] According to some embodiments of the invention, the structure of the receptor compound sets the absolute structure of Ring I and Ring II in the resulting compound.

[0775] The Examples section and Figures below show exemplary receptors suitable for preparing compounds of Formula III*a (see e.g. Fig.15 and 16 ).

[0776] In the context of methods of preparing the compounds of the present embodiments, embodiments of the present invention further encompass any intermediate compounds as described herein.

[0777] Therapeutic Uses:

[0778] As is known in the art, about one-third of alleles causing genetic diseases carry premature termination (termination) (stop) codons (PTCs), resulting in the production of truncated proteins. A possible treatment method involves inducing and / or promoting the readout of such PTCs to enable the synthesis of full-length proteins. PTCs originate from mutations, such as nonsense mutations, frameshift deletions and insertions, or from abnormal splicing of mRNA isoforms that produce truncated reading frames. Due to major negative effects or gain-of-function effects, these mutations can result in the production of truncated, non-functional or harmful proteins.

[0779] Typically, readout of PTCs can be achieved by suppressor transfer RNAs (tRNAs), factors that reduce translation termination efficiency, such as small interfering RNAs (siRNAs) against translation termination factors, and RNA antisense targeting nonsense mutation regions. It is an object of the present invention to provide a pharmacological treatment method, the purpose of which is to achieve sufficient levels of functional protein in a subject suffering from at least one genetic disorder associated with at least one premature stop codon mutation.

[0780] According to some embodiments of the invention, provided therapeutic methods are directed to inducing and / or promoting the translational readout of the disease causing PTC, so as to enable the synthesis and expression of a full-length functional protein.

[0781] As described above, a method that has been extensively studied has been carried out to clinical trials, which is based on the readout performed using drugs (such as aminoglycoside antibiotics) that affect ribosomal decoding sites. However, when used in high concentrations and / or used for a long time, aminoglycosides have serious adverse side effects. The compounds shown herein are designed to show the ability to induce and / or promote the readout of premature termination codon mutations in organisms with such mutations, while showing minimal adverse effects. This activity makes these compounds suitable for use as therapeutic agents for the treatment of genetic disorders associated with premature termination codon mutations.

[0782] As described in the Examples section below, it is shown that the exemplary compounds covered by the present embodiment do exhibit premature stop codon mutation inhibitory activity and can be used to induce the readout of genes characterized by pathogenic premature stop codon mutations, and thus play a role in treating corresponding genetic diseases or disorders associated with premature stop codon mutations.

[0783] Another recently proposed approach to inhibiting stop codon mutations (PTCs) involves attenuating nonsense-mediated mRNA decay (NMD), a conserved eukaryotic cell pathway that targets PTC-containing mRNAs for degradation. Attenuation of NMD has been shown to increase the abundance of PTC-containing mRNAs and thereby restore higher levels of functional protein produced by PTC inhibition [see, e.g., Keeling et al., PLoS ONE 8(4): e60478, 2013; Bidou et al., RNA Biology 14(3): 378-388, 2017]. Additional studies have shown that aminoglycosides (e.g., NB124) described in, for example, WO 2012 / 066546 exhibit attenuation of NMD, suggesting that the aminoglycoside compounds of the present embodiments may also attenuate NMD [see, e.g., Alroy et al., Abstracts / Molecular genetics and metabolism 2018, 123(2): S18].

[0784] According to one aspect of some embodiments of the present invention, any compound shown herein, for example, having formula A, B, I, I*, III, III*, IV or IVa, preferably formula A, B, I, I*, III or III*, including any corresponding embodiment of the compound and any combination thereof (as well as compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) is used to reduce nonsense-mediated mRNA decay (NMD), and / or for the manufacture of drugs that reduce nonsense-mediated mRNA decay (NMD), and / or for the treatment of diseases or disorders associated with unregulated (irregulated) nonsense-mediated mRNA decay (NMD), and / or diseases or disorders that can be treated by reducing nonsense-mediated mRNA decay (NMD). In some embodiments, the disease or disorder is a genetic disease or disorder as described herein in any corresponding embodiment. In some embodiments, the disease or disorder is a cancer as described herein in any corresponding embodiment.

[0785] According to one aspect of some embodiments of the present invention, any compound shown herein, for example having formula A, B, I, I*, III, III*, IV or IVa, preferably formula A, B, I, I*, III or III*, including any corresponding embodiments of the compound and any combination thereof (as well as compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) is used to treat cancer as defined herein, or for the manufacture of a drug for treating cancer as defined herein, or for a method of treating cancer as defined herein. According to some embodiments, the compounds described herein are used to induce and / or promote the readout of premature stop codon (nonsense) mutations in tumor suppressor genes (e.g., p53). According to some embodiments, the compounds described herein are used to treat cancer by attenuating NMD.

[0786] According to one aspect of some embodiments of the present invention, any compound shown herein, for example having formula A, B, I, I*, III, III*, IV or IVa, preferably formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof (as well as including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) is used to induce and / or promote the readout of premature stop codon mutations and / or for increasing the expression of genes having premature stop codon mutations, and / or for the manufacture of a drug for inducing and / or promoting the readout of premature stop codon mutations and / or for increasing the expression of genes having premature stop codon mutations.

[0787] According to one aspect of some embodiments of the present invention, any compound shown herein, for example having formula A, B, I, I*, III, III*, IV or IVa, preferably formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof (as well as including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) is used to treat a genetic disorder associated with a premature stop codon mutation, or for the manufacture of a medicament for the treatment of a genetic disorder associated with a premature stop codon mutation.

[0788] Any premature stop codon mutation is contemplated. In some embodiments, the mutations are those with RNA codons of UGA, UAG or UAA.

[0789] According to some of any of the embodiments described herein, the protein is translated in a cytoplasmic translation system.

[0790] According to some of any of the embodiments described herein, the compound is used in a mutation inhibiting amount.

[0791] According to some of any of the embodiments described herein, the translation inhibition IC of the compound in the cytoplasmic translation system of a eukaryotic organism 50 Greater than the IC of translation inhibition of the compound in the ribosomal translation system 50 .

[0792] According to some of any of the embodiments described herein, the translation inhibition IC of the compound in the cytoplasmic translation system of a eukaryotic organism 50 Greater than the IC of translation inhibition of the compound in a prokaryotic translation system 50 .

[0793] According to one aspect of some embodiments of the present invention, any compound shown herein, for example having formula A, B, I, I*, III, III*, IV or IVa, preferably formula A, B, I, I*, III or III*, including any corresponding embodiments of the compounds and any combination thereof (as well as including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b) is used to treat a genetic disorder associated with a premature stop codon mutation, or for the manufacture of a medicament for the treatment of a genetic disorder associated with a premature stop codon mutation.

[0794] According to one aspect of some embodiments of the present invention, a method for treating a genetic disorder associated with a premature stop codon mutation is provided. The method according to this aspect of the present invention is achieved by administering a therapeutically effective amount of one or more compounds proposed herein to a subject in need thereof, for example, having formula A, B, I, I*, III, III*, IV or IVa, preferably formula A, B, I, I*, III or III*, including any corresponding embodiment of the compound and any combination thereof (as well as including compounds represented by formula Ia, I**, I*a, I*b, IIIa, III**, III*a, and III*b).

[0795] As used herein, the term "treating" includes abrogating, substantially inhibiting, slowing, or reversing the progression of a condition, substantially ameliorating the clinical or aesthetical symptoms of a condition, or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.

[0796] As used herein, the phrase "therapeutically effective amount" describes an amount of the polymer being administered that will relieve to some extent one or more of the symptoms of the condition being treated.

[0797] As used herein, the phrase "genetic disorder" refers to a chronic disorder caused by one or more defective genes that are often inherited from both parents and occur unexpectedly when two healthy carriers of a defective recessive gene reproduce or when the defective gene becomes dominant. Genetic disorders can occur in different inheritance patterns, including autosomal dominant inheritance patterns, in which only one mutant copy of a gene is required to affect offspring, and autosomal recessive inheritance patterns, in which mutations must occur in both copies of a gene to affect offspring.

[0798] As used herein, the phrase "genetic disorder" encompasses a genetic disorder, a genetic disease, a genetic condition, or a genetic syndrome.

[0799] According to some of any embodiments of the present invention, genetic disorders, genetic diseases, genetic conditions, or genetic syndromes relate to genes with premature stop codon mutations (also referred to herein as truncated mutations and / or nonsense mutations) that cause improper translation thereof. Improper translation can produce dysfunctional essential proteins, or cause a reduction or cancellation of essential protein synthesis. In the context of some embodiments of the present invention, the genetic disorders considered within the scope of this embodiment are referred to as genetic disorders associated with premature stop codon mutations and / or protein truncation phenotypes.

[0800] According to some of any embodiments of the invention, genetic disorders associated with premature stop codon mutations and / or protein truncation phenotypes can be treated by inducing and / or promoting the readout of mutations in intact but defective transcripts (mRNA), or in other words, by inducing and / or promoting the suppression of nonsense mutations (premature stop codon mutations and / or truncating mutations). In the context of embodiments of the invention, therefore, a genetic disorder is a disorder that can be treated by a compound that induces and / or promotes readout.

[0801] Methods for identifying genetic disorders associated with premature stop codon mutations and / or protein truncation phenotypes are well known in the art and include full or partial genome elucidation, genetic biomarker testing, phenotypic classification, and genetic information analysis.

[0802] Such methods typically result in pairs of mutant / wild-type (WT) sequences, and these pairs can be used in known methods to identify whether a genetic disorder is associated with a premature stop codon mutation and / or a protein truncation phenotype.

[0803] The readout inducing and / or promoting activity of compounds useful in treating such genetic disorders can be established by methods well known in the art.

[0804] For example, in a whole cell or cell-free system, a plasmid containing two reporter genes interrupted by a mutant gene (a gene causing a genetic disorder) sequence is transected into a protein expression platform, and the ratio between the expression levels of the two genes in the presence of a test compound is measured, usually over a series of concentrations and duplications, and compared with the ratio of the gene expression levels of the wild type and / or with the ratio of the expression levels measured in a control sample that does not contain the test compound.

[0805] It is noteworthy that the experimental model used for readout activity, i.e., the nucleotide sequence of a gene containing a premature stop codon mutation, is a byproduct of a process for identifying genetic disorders associated with premature stop codon mutations and / or protein truncation phenotypes, and it is also noteworthy that with the tremendous advances in genomic data acquisition, this process is now well within the skill of those skilled in the art, and once the mechanism of action of a candidate drug is established, such as in the case of a genetic disorder that has been demonstrated to be associated with a premature stop codon mutation and / or protein truncation phenotype, it is within the skill of those skilled in the art to identify, characterize and evaluate the efficacy, selectivity and safety of any of the readout-inducing compounds presented herein. It is also within the skill of those skilled in the art to further employ the readout-inducing compounds presented herein throughout the routine process of drug development.

[0806] Methods for testing the readout of premature stop codon mutations and / or truncated mutations, referred to herein as readout activity, are known in the art, and several exemplary experimental methods are provided in the Examples section below, whereby compounds induced by readout according to some embodiments of the present invention can be characterized. It should be understood that other methods can be used to characterize compounds induced by readout, and these methods are also contemplated within the scope of the present invention. The methods provided herein can also be applied to high-throughput screening techniques, which can evaluate thousands of compounds in a relatively short period of time.

[0807] Those skilled in the art will appreciate that many in vitro methods can be used to characterize the safety of the readout-inducing compounds provided herein when used as drugs, and to assess the cytotoxicity of candidate drugs relative to their efficacy. The skilled person will also appreciate that many in vitro methods can be used to characterize the selectivity of the readout-inducing compounds provided herein for eukaryotic versus prokaryotic organisms, and that such methods can also be adapted for use in high-throughput screening techniques that can evaluate thousands of compounds in a relatively short period of time.

[0808] Non-limiting examples of genetic disorders, diseases, conditions, and syndromes associated with the presence of at least one premature stop codon or other nonsense mutation include cancer, Rett syndrome, cystic fibrosis (CF), Becker muscular dystrophy (BMD), congenital muscular dystrophy (CMD), Duchenne muscular dystrophy (DMD), factor VII deficiency, familial atrial fibrillation, Hailey-Hailey disease, hemophilia A, hemophilia B, Hurler syndrome, Louis-Bar syndrome (ataxia-telangiectasia, AT), McArdle disease, mucopolysaccharidoses, nephrotic cystinosis, polycystic kidney disease, spinal muscular atrophy type I, II, III (SMA), Tay-Sachs, Usher syndrome, cystinosis, severe epidermolysis bullosa, Dravet syndrome, X-linked nephrotic diabetes insipidus (XNDI), and X-linked retinitis pigmentosa.

[0809] Kim M. Keeling, KM Bedwell, DM, “Suppression of nonsense mutations as a therapeutic approach to treat genetic diseases” in Wiley Interdisciplinary Reviews: RNA, 2011, 2(6), pp.837-852; Bordeira- Other genetic disorders, diseases, conditions, and syndromes associated with the presence of at least one premature stop codon or other nonsense mutation are listed in "Cancer syndromes and therapy by stop-codon readthrough" in Trends in Molecular Medicine, 2012, 18(11), p. 667-678, and in any reference cited herein, all of which are incorporated herein by reference in their entirety.

[0810] As used herein, the terms "cancer," "cancerous disease," and "tumor" are used interchangeably. The terms refer to malignant growths and / or tumors caused by abnormal and uncontrolled cell proliferation (cell division). The term "cancer" encompasses tumor metastases. The term "cancer cell" describes cells that form malignant growths or tumors.

[0811] Non-limiting examples of cancer and / or tumor metastasis include any solid or non-solid cancer and / or tumor metastasis, including, but not limited to, gastrointestinal tumors (e.g., colon cancer, rectal cancer, colorectal carcinoma, colorectal cancer, colorectal adenoma, hereditary nonpolyposis type 1, hereditary nonpolyposis type 2, hereditary nonpolyposis type 3, hereditary nonpolyposis type 6; colorectal cancer, hereditary nonpolyposis type 7, small and / or large intestine cancer, esophageal cancer, tylosis with esophageal cancer, gastric cancer, pancreatic cancer, pancreatic endocrine tumors), endometrial cancer, dermatofibrosarcoma protuberans, gallbladder cancer, biliary tract tumors, prostate cancer, prostate adenocarcinoma, kidney cancer (e.g., Wilms' tumor type 2 or type 1), liver cancer (e.g., hepatoblastoma, hepatocellular carcinoma, hepatocellular carcinoma cancer), bladder cancer, embryonal rhabdomyosarcoma, germ cell tumor, trophoblastic tumor, testicular germ cell tumor, ovarian immature teratoma, uterus, ovarian epithelial cancer, sacrococcygeal tumor, choriocarcinoma, placental site trophoblastic tumor, adult epithelial tumor, ovarian cancer, serous ovarian cancer, ovarian sex cord tumor, cervical cancer, cervical cancer, small cell and non-small cell lung cancer, nasopharyngeal carcinoma, breast cancer (e.g., ductal breast cancer, invasive intraductal breast cancer, sporadic breast cancer, predisposition breast cancer, breast cancer type 4, breast cancer-1, breast cancer-3, breast-ovarian cancer), squamous cell carcinoma (e.g., head and neck), neurogenic tumors, astrocytoma, ganglioblastoma, neuroblastoma, lymphoma (e.g., Hodgkin's disease, non-Hodgkin's lymphoma, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), Burkitt's lymphoma, cutaneous T-cell lymphoma, histiocytic lymphoma, lymphoblastic lymphoma, T-cell lymphoma, thymic lymphoma, tumours), gliomas, adenocarcinomas, adrenal gland tumours, hereditary adrenocortical carcinomas, brain malignancies (tumours), various other carcinomas (e.g. bronchial large cell carcinoma, ductal carcinoma, Ehrlich's ascites carcinoma, epidermoid carcinoma, large cell carcinoma, Lewis lung carcinoma, medullary carcinoma, mucoepidermoid carcinoma, oat cell carcinoma, small cell carcinoma, spindle cell carcinoma, spinocerebellar carcinoma, transitional cell carcinoma, undifferentiated carcinoma, carcinosarcoma, choriocarcinoma, cystadenocarcinoma), epithelial cell tumours, epithelioma, erythroleukaemias (e.g. , Friend, lymphoblastic), fibrosarcoma, giant cell tumor, glial tumor, glioblastoma (e.g., multiforme, astrocytoma), glioma hepatocellular carcinoma, heterohybridoma, hybrid myeloma, histiocytoma, hybridoma (e.g., B cell), adrenal tumor, insulinoma, islet tumor, keratoma, leiomyoma, leiomyosarcoma, leukemia (e.g., acute lymphoblastic leukemia, acute lymphoblastic leukemia, acute lymphoblastic pre-B cell leukemia,Acute lymphocytic T-cell leukemia, acute megakaryocytic leukemia, monocytic leukemia, acute myeloid leukemia, acute myeloid leukemia, acute myeloid leukemia with eosinophilia, B-cell leukemia, basophilic leukemia, chronic myeloid leukemia, chronic B-cell leukemia, eosinophilic leukemia, Friend's leukemia, granulocytic or myeloid leukemia, hairy cell leukemia, lymphocytic leukemia, megakaryoblastic leukemia, monocytic leukemia, mononuclear macrophage Myeloblastic leukemia, myeloblastic leukemia, myelomonocytic leukemia, plasma cell leukemia, pre-B cell leukemia, promyelocytic leukemia, subacute leukemia, T-cell leukemia, lymphoid tumors, prone to myeloid malignancies, acute non-lymphoid leukemia), lymphosarcoma, melanoma, breast tumors, mast cell tumors, medulloblastoma, mesothelioma, metastatic tumors, monocytic tumors, multiple myeloma, myelodysplastic syndrome, myeloma, Wilms tumor, neural tissue glioma, neuroblastoma Histioneuroma, Schwannoma, Neuroblastoma, Oligodendroglioma, Osteochondroma, Myeloma, Osteosarcoma (e.g., Ewing's), Papilloma, Transitional cell, Pheochromocytoma, Pituitary tumor (invasive), Plasmacytoma, Retinoblastoma, Rhabdomyosarcoma, Sarcomas (e.g., Ewing's, Histiocytic, Jensen, Osteogenic, Reticular cell), Schwannoma, Subcutaneous tumors, Malignant malformations (e.g., Multipotent), Teratoma, Testicular tumors, Thymoma and Trichoepithelioma, Gastric cancer, Fibrosarcoma , glioblastoma multiforme, multiple glomus tumors, Li-Fraumeni syndrome, liposarcoma, lynch cancer family syndrome II, male germ cell tumor, mast cell leukemia, medullary thyroid, multiple meningiomas, endocrine adenoma myxosarcoma, paraganglioma, familial non-phenotypic cytoma, pilomatricoma, papillary carcinoma, familial versus sporadic carcinoma, rhabdoid predisposition syndrome, familial carcinoma, rhabdoid tumor, soft tissue sarcoma, and Turcot syndrome with glioblastoma. ,

[0812] In any of the methods and uses described herein, the compounds described herein may be used per se or form part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier as defined herein.

[0813] According to an aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising as an active ingredient any novel compound described herein and a pharmaceutically acceptable carrier.

[0814] As used herein, "pharmaceutical composition" refers to a preparation of a compound presented herein with other chemical components such as pharmaceutically acceptable and suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0815] Hereinafter, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. Examples of carriers include, but are not limited to, propylene glycol, saline, emulsions and mixtures of organic solvents and water, and solid (e.g., powdered) and gaseous carriers.

[0816] The term "excipient" herein refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of a compound. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0817] Techniques for formulation and administration of drugs may be found in the most recent edition of "Remington's Pharmaceutical Sciences", Mack Publishing Co., Easton, PA, which is incorporated herein by reference.

[0818] The pharmaceutical composition of the present invention can be manufactured by methods well known in the art, for example, by conventional mixing, dissolving, granulating, dragee-making, leaching, emulsifying, encapsulating, embedding or lyophilizing methods.

[0819] Thus, the pharmaceutical compositions used according to the present invention can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the compounds set forth herein into pharmaceutically usable preparations. Suitable formulations depend on the chosen route of administration.

[0820] According to some embodiments, the administration is completed by oral administration. For oral administration, the compounds provided herein can be easily prepared by mixing the compounds with pharmaceutically acceptable carriers known in the art. This carrier enables the compounds proposed herein to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by patients. Solid excipients can be used, the resulting mixture is optionally ground, and the granular mixture is processed after adding suitable adjuvants when necessary to prepare tablets or dragee cores, thereby preparing pharmaceutical preparations for oral administration. Suitable excipients are especially fillers, such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose; and / or physiologically acceptable polymers, such as polyvinyl pyrrolidone (PVP). If necessary, disintegrants such as cross-linked polyvinyl pyrrolidone, agar, or alginic acid or its salts, such as sodium alginate, can be added.

[0821] Pharmaceutical compositions that can be used orally include push-fit capsules made of gelatin and sealed soft capsules made of gelatin and plasticizers (such as glycerol or sorbitol). Push-fit capsules can contain active ingredients mixed with fillers (such as lactose), binders (such as starches), lubricants (such as talc or magnesium stearate), and optional stabilizers. In soft capsules, the compounds presented herein can be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers can be added. The dosage of all preparations for oral administration should be suitable for the selected route of administration.

[0822] For injection, the compounds presented herein can be formulated in aqueous solutions, preferably in physiologically compatible buffers (eg, Hank's solution, Ringer's solution, or physiological saline buffer) with or without organic solvents (eg, propylene glycol, polyethylene glycol).

[0823] For transmucosal administration, penetrants are used in the formulation. Such penetrants are well known in the art.

[0824] The dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol, titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablet or dragee coating to identify or characterize different combinations of active aminoglycoside compound dosages.

[0825] For oral administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0826] For administration by inhalation, the compounds presented herein are conveniently delivered in the form of an aerosol spray delivery (generally including a powdered, liquid, and / or gaseous carrier) from a pressurized pack or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, or carbon dioxide). In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of a compound presented herein and a suitable powder base (e.g., but not limited to, lactose or starch).

[0827] The compounds presented herein can be formulated for parenteral administration, for example by bolus injection or continuous infusion. Injectable formulations can be in unit dosage form, for example in ampoules or in multi-dose containers, optionally with the addition of preservatives. The composition can be a suspension, solution, or emulsion in an oily or aqueous vehicle, and can contain preparatants, such as suspending agents, stabilizers, and / or dispersants.

[0828] Pharmaceutical compositions for parenteral administration include aqueous solutions of compound preparations in water-soluble form. In addition, the suspension of the compound proposed herein can be prepared into suitable oily injection suspensions and emulsions (e.g., water-in-oil, oil-in-water, or water-in-oil in oil emulsions). Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate, triglycerides, or liposomes. Aqueous injection suspensions may include substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also include a suitable stabilizer or reagent that increases the solubility of the compound proposed herein to allow the preparation of a high concentration solution.

[0829] Alternatively, the compounds presented herein may be in powder form for mixing with a suitable vehicle (eg, sterile, pyrogen-free water) before use.

[0830] The compounds presented herein may also be formulated in rectal compositions such as suppositories or retention enemas, using, eg, conventional suppository bases such as cocoa butter or other glycerides.

[0831] The pharmaceutical compositions described herein may also include a suitable solid gel phase carrier or excipient. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers, such as polyethylene glycol.

[0832] Pharmaceutical compositions suitable for use in the context of the present invention include compositions containing active ingredients in an amount effective to achieve the intended purpose. More specifically, a therapeutically effective amount refers to an amount of the compound presented herein that is effective to prevent, alleviate or improve symptoms of a disorder, or prolong the survival of the treated subject.

[0833] Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0834] For any compound presented herein used in the methods of the present embodiment, the therapeutically effective amount or dosage can be first estimated from an activity assay in an animal. For example, a dose can be formulated in an animal model to achieve a circulating concentration range (e.g., a concentration of a test compound that achieves a basic readout of a truncated mutation) that includes the level of mutation inhibition determined by an activity assay. Such information can be used to more accurately determine a dose useful for humans.

[0835] The results can be obtained by standard pharmaceutical procedures in experimental animals, for example by determining the EC 50 (the concentration of the compound at which 50% of its maximal effect is observed) and LD 50(lethal dose, resulting in death of 50% of the test animals), to determine the toxicity and therapeutic efficacy of the compounds presented herein. The data obtained from these activity assays and animal studies can be used to formulate a dosage range for use in humans.

[0836] The dosage may vary depending on the dosage form employed and the route of administration employed. The specific formulation, route of administration, and dosage may be selected by the individual physician based on the patient's condition. (e.g., Fingl et al., 1975, in "The Pharmacological Basis of Therapeutics", Chapter 1, page 1).

[0837] Dosage and interval can be adjusted individually to provide a plasma level of the compound presented herein sufficient to maintain the desired effect, referred to as the minimum effective concentration (MEC). The MEC of each preparation will vary, but can be estimated based on in vitro data; for example, the concentration of the compound necessary to achieve 50-90% expression of the entire gene with a truncated mutation (i.e., the readout of the mutant codon). The dosage required to reach the MEC will depend on individual characteristics and route of administration. HPLC assays or bioassays can be used to determine plasma concentrations.

[0838] Dosage intervals can also be determined using the MEC. Formulations should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably 30-90% of the time, and most preferably 50-90% of the time.

[0839] Depending on the severity and responsiveness of the chronic disease to be treated, administration may also be a single cyclical administration of the above-described sustained-release composition, wherein the course of cyclical treatment lasts for several days to several weeks, or until sufficient improvement is achieved during the cyclical treatment, or until a significant reduction in the disorder can be achieved during the cyclical treatment.

[0840] Of course, the amount of the composition to be administered depends on the subject being treated, the severity of the disease, the mode of administration, the judgment of the prescribing physician, etc. The composition of the present invention can be provided in the form of a package or dispenser device (e.g., a kit approved by the FDA (U.S. Food and Drug Administration)) if necessary, which may include one or more unit dosage forms containing active ingredients. The packaging can, for example, include metal or plastic film, such as but not limited to blister packaging or pressurized containers (for inhalation). The packaging or dispenser device can be accompanied by instructions for use. The packaging or dispenser may also be accompanied by a reminder associated with the container, which is a form specified by a government agency that regulates drug production, use or sales, and the notice reflects the agency's approval of the form of human or veterinary compositions. For example, such a notice can be a prescription drug label approved by the U.S. Food and Drug Administration, or it can be an approved product insert. As described in detail above, a composition of a compound according to the present embodiment formulated in a compatible drug carrier can be prepared and placed in a suitable container and marked for the treatment of a specified illness or diagnosis.

[0841] Thus, in some embodiments, the pharmaceutical composition is packaged in packaging material and is labeled in printed form in or on the packaging material for use in treating a genetic disorder as defined herein, and / or for any of the uses described herein.

[0842] In some embodiments, the pharmaceutical composition is for use in treating a genetic disorder as defined herein, and / or for any of the uses described herein.

[0843] In any of the compositions, methods, and uses described herein, the compounds can be used in combination with other agents for treating genetic disorders, as described herein, and / or for inducing or promoting the readout activity of premature stop codon mutations and / or for increasing the expression of genes having premature stop codon mutations.

[0844] Exemplary such agents include, but are not limited to, CFTR potentiators, such as ivacaftor (VX-770) (see X. Xue et al., Am. J. Respir. Cell Mol. Biol. 50 (4), 805-816 (2014)); and agents that reduce nonsense-mediated mRNA decay (NMD), such as NMDI-1, caffeine and other agents that disrupt the UPF1 phosphorylation cycle (see KMKeeling et al., PLoS ONE 8 (4), e60478 (2013)). Any other agent is also contemplated.

[0845] The compounds or pharmaceutical compositions comprising the compounds proposed in the present invention are mainly aimed at the treatment of genetic disorders defined as chronic diseases, and it is expected that they will be administered throughout the life cycle of the subject being treated. Therefore, the mode of administration of the pharmaceutical composition containing the compound should make the administration easy and comfortable, preferably self-administered, so as to minimize the loss of the patient's health and life cycle.

[0846] Repeated and periodic administration of the compounds presented herein or pharmaceutical compositions containing the compounds can be, for example, performed on a daily basis, i.e., once a day, more preferably, the administration is performed on a weekly basis, i.e., once a week, more preferably, the administration is performed on a monthly basis, i.e., once a month, and most preferably, once every several months (e.g., once every 1.5 months, 2 months, 3 months, 4 months, 5 months, or even 6 months).

[0847] As discussed above, some of the limitations to using currently known aminoglycosides as readout drugs for truncated mutations are associated with the fact that they are primarily antibacterial (used as antibiotics). Long-term use of any antibacterial agent is difficult to guarantee safety and can even be life-threatening because it alters the intestinal microbial flora, which may cause or aggravate other medical conditions, such as flare-ups of inflammatory bowel disease, and may lead to resistance in certain pathological microbial strains.

[0848] In some embodiments, the compounds presented herein have substantially no antibacterial activity. "No antibacterial activity" means that the minimum inhibitory concentration (MIC) for a particular strain is much higher than the concentration of the compound that is considered to have an antibiotic effect on the strain. In addition, the MIC of these compounds is significantly higher than the concentration required to exert the inhibitory activity of the truncated mutation.

[0849] The compounds proposed herein are essentially non-bactericidal and do not produce the above-mentioned adverse effects, and can therefore be administered via absorption routes that may contain non-targeted benign and / or beneficial microorganisms, and therefore may even need to be retained. This important property of the compounds proposed herein makes these compounds particularly effective in combating chronic diseases, as they can be administered repeatedly and over the lifespan without causing any adverse cumulative effects associated with antimicrobials, and can further be administered orally or rectally, i.e., through the gastrointestinal tract (CI tract), a very useful and important property for drugs treating chronic disorders.

[0850] According to some embodiments, the compounds proposed herein are selected and / or designed to be selective for the prokaryotic translation system relative to the eukaryotic translation system, i.e., the compound exhibits higher activity in eukaryotic cells (e.g., in mammals (humans)) compared to the activity in prokaryotic cells (e.g., bacteria). Without being bound by any particular theory, it is believed that the compounds proposed herein are known to act by binding to the A site of 16S ribosomal RNA, and the ribosome is involved in the translation of genes, so that the compounds proposed by the present invention have a higher affinity for the eukaryotic ribosome A site, or are selective for the eukaryotic A site relative to the prokaryotic ribosome A site and the mitochondrial ribosome A site similar to its prokaryotic counterpart.

[0851] As used herein, the term "about" refers to ± 10%.

[0852] The terms "comprises," "comprising," "includes," "including," "having" and their conjugations mean "including but not limited to."

[0853] The term "consisting of" means "including and limited to."

[0854] The term "consisting essentially of" means that the composition, method, or structure may include additional ingredients, steps, and / or parts, but only if the additional ingredients, steps, and / or parts do not materially change the basic and novel characteristics of the claimed composition, method, or structure.

[0855] As used herein, the singular forms "a", "an", and "the" include plural references unless the context clearly indicates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof. Throughout the application, various embodiments of the present invention may be presented in a range format. It should be understood that the description in range format is for convenience and brevity only and should not be interpreted as an inflexible limitation on the scope of the present invention. Therefore, the description of the range should be considered to have specifically disclosed all possible sub-ranges and individual numerical values ​​within the range. For example, the description of a range from 1 to 6 should be considered to have explicitly disclosed sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual numbers within the range, such as 1, 2, 3, 4, 5, and 6. This has nothing to do with the breadth of the range.

[0856] When a numerical range is indicated herein, it is intended to include any cited numeral (fractional or integer) within the indicated range. The phrases "range / range between" a first indicated numeral and a second indicated numeral and "range / range from" a first indicated numeral "to" a second indicated numeral are used interchangeably herein and are intended to include the first and second indicated numerals and all fractions and integers therebetween.

[0857] As used herein, the term "method" refers to manners, means, techniques, and steps for accomplishing a given task, including but not limited to those manners, means, techniques, and steps known to practitioners in the chemical, pharmacological, biological, biochemical, and medical fields or readily developable from known manners, means, techniques, and steps.

[0858] It is expected that within the term of this patent application, many related genetic diseases and disorders will be discovered, and the scope of the term is intended to include a priori all such new disorders and diseases.

[0859] It should be understood that certain features of the invention described in the context of separate embodiments for clarity may also be provided in combination in a single embodiment. Conversely, various features of the invention described in the context of a single embodiment for brevity may also be provided individually or in any suitable subcombination or in any other described embodiment of the invention as appropriate. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiment cannot function without those elements.

[0860]

[00136] Various embodiments and aspects of the present invention as delineated hereinabove and in the claims section below find experimental support in the following examples. Example

[0861] Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non-limiting fashion.

[0862] Materials and methods

[0863] On Bruker AvanceTM 500 / 400 (excluding 1D TOCSY) / 200 (only 1 H NMR) spectrometer 1 H-NMR, 13 C-NMR, DEPT, COSY, 1D TOCSY, HMQC, and HMBC Unless otherwise stated, chemical shifts reported (in ppm) are relative to internal Me4Si in CDCl3 (δ = 0.0) and HOD in D2O (δ = 4.63).

[0864] Mass spectrometric analyses were performed on a Bruker Daltonix Apex 3 mass spectrometer under electron spray ionization (ESI), on a TSQ-70B mass spectrometer (Finnigan Mat) or on MALDI-TOF on an α-cyano-4-hydroxycinnamic acid matrix on a MALDI Micromass mass spectrometer.

[0865] The reaction was monitored by TLC on silica gel 60 F254 (0.25 mm, Merck) and by using a 10% H2SO4 solution containing (NH4)6Mo7O 24 • Carbonization of yellow solution of 4H2O (120 gr) and (NH4)2Ce(NO3)6 (5 gr) was used to visualize the spot.

[0866] Flash column chromatography was performed on silica gel Gel 60 (70-230 mesh).

[0867] Unless otherwise stated, all reactions were carried out using anhydrous solvents.

[0868] Unless otherwise stated, all chemicals were obtained from commercial sources.

[0869] Example 1

[0870] Chemical Synthesis of Some Exemplary N1-Substituted Compounds According to the Present Embodiments

[0871] A library of newly designed aminoglycoside derivatives featuring acyl and sulfinyl groups as substituents at the N1 position was prepared. The initial library was based on the previously described lead candidate NB74 (see Table 1 ) with modifications at the N1 position (see Figure 2 , Group 1) such as acetate (NB74-N1Ac), benzoate (NB74-N1Bz), methanesulfonate (NB74-N1MeS) and phenylsulfonate (NB74-N1PhS). Then, similar modifications were made at the N1 position of the previously described lead candidate NB124 (see Table 1) to give compounds NB124-N1Ac, NB124-N1Bz and NB124-N1MeS (see Figure 2 , group 2).

[0872] according to Figure 3 The general synthetic routes shown prepare exemplary newly designed compounds.

[0873] In brief, commercially available G418 was converted into a known common intermediate A in four chemical steps according to the previously reported synthetic steps (see Nudelman et al., Bioorg. Med. Chem. 18, 3735-46 (2010)). Intermediate A was then acetylated or sulfonated at the free amine (N1 position) and then selectively acetylated to provide a series of common acceptors B, wherein N1 was modified with different amide or sulfonamide moieties, and the free hydroxyl group at the C-5 position was ready for coupling of ring III when needed. The coupling step of ring III was carried out according to the previously reported synthetic steps, using the corresponding trichloroacetimidate donor, followed by two steps of deprotection to obtain the desired group 1 and group 2 structures.

[0874] The following are methods for preparing exemplary compounds according to some embodiments of the present invention, which are described in Tables 1 and 2 above. Figure 2 Shown in.

[0875] Figure 4 The synthetic pathway for the conversion of G418 to acceptor compounds 5, 6, 7 and 9 is shown.

[0876] Figure 5 The synthetic pathways for converting receptors 5-7 and 9 to the corresponding N1-modified NB-74 and NB-124 are shown.

[0877] 2′,3-diazide-1-N-benzamide-6′-(R)-methyl-paromomycinamine (compound 3; Figure 4 ):

[0878]

[0879] Compound 2 (2 g, 5.13 mmol), prepared as described in Nudelman, I. et al. Bioorganic Med. Chem. 18, 3735-3746 (2010), was dissolved in anhydrous pyridine (10 ml). The stirred mixture was cooled to 0°C on an ice bath, and then benzoyl chloride (12 eq., 0.061 mol, 7 ml) was added. The propagation of the reaction was monitored by TLC (EtOAc / hexane 3:7). The mixture was diluted with EtOAc and washed with HCl 1M, NaHCO3 (saturated) and brine. The combined organic layers were dried over MgSO4, and the mixture was evaporated to dryness and treated with MeNH2 (33% solution in EtOH). The propagation of the reaction was monitored by TLC (EtOAc / MeOH, 1:1), indicating completion after 12 hours. The reaction mixture was evaporated to dryness and purified by silica gel chromatography (100% EtOAc) to produce compound 3 (2 g, 79%).

[0880] 1H NMR (500MHz, MeOD): Ring I: δ=5.77 (d, 1H, J=3.5Hz, H-1), 4.06 (dd, 1H, J=6.8, 3.6Hz, H-6), 4.02-3.93 (m, 2H, H-3, H-5), 3.40 ( dd, 1H, J=10.0, 8.7Hz, H-4), 3.11 (dd, 1H, J=10.6, 4.2Hz, H-2), 1.28 (d, 3H, J=6.1Hz, CH3-C-6). Ring II: δ=4.04 (ddd, 1H, J=11. 9, 10.1, 4.4 Hz, H-1), 3.66-3.54 (m, 3H, H-3, H-6, H-5), 3.45 (dd, 1H, J = 10.1, 9.0 Hz, H-4), 2.34-2.29 (m, 1H, H-2), 1.58 (dd, 1H, J = 25.1, 12.3 Hz, H-2). The additional peaks in the spectrum were identified as follows: δ = 7.85 (d, J = 7.2 Hz, 2H, Ph), 7.53 (t, J = 7.4 Hz, 1H, Ph), 7.45 (t, J = 7.6 Hz, 2H, Ph).

[0881] 13 C NMR (126MHz, MeOD): δ = 169.02 (carbonyl), 134.28 (Ph), 131.24 (Ph), 128.04 (Ph), 126.98 (Ph), 97.31 (C-1′), 78.93 (C-4), 77.31 (C-5), 74.59 ( C-6), 73.76(C-3′), 72.83(C-4′), 70.91(C-5′), 67.91(C-1), 63.26(C-2′), 60.11(C-3), 49.70(C-6′), 32.45(C-2), 16.58(CH3-C-6′).

[0882] MALDI TOF MS: C 20 H27N7O8([M+H] + )m / e 493.47; measured value is m / e 494.2.

[0883] 2′,3-diazide-1-N-methanesulfonamide-6′-(R)-methyl-paromomycinamine (compound 4; Figure 4 ) Preparation:

[0884]

[0885] Compound 2 (2.5 g, 6.42 mmol) prepared as described by Nudelman, I. et al. 2010 (supra) was dissolved in DMF, then methanesulfonyl chloride (1 eq., 0.5 ml) was added, and the resulting mixture was stirred at room temperature for 24 hours. The propagation of the reaction was monitored by TLC (MeOH / EtOAc 1:9). The mixture was then evaporated in vacuo and purified by silica gel chromatography. The product was eluted with 100% EtOAc. The fractions containing the product were combined and evaporated in vacuo to give compound 4 (1 g, 33%).

[0886] 1 H NMR (500MHz, MeOD): Ring I: δ=5.70 (d, 1H, J=3.5Hz, H-1), 4.04-4.00 (m, 1H, H-6), 3.96-3.88 (m, 2H, H-3, H-5 ), 3.35 (t, 1H, J=9.5Hz, H-4), 3.07 (dd, 1H, J=10.4, 4.0Hz, H-2), 1.24 (d, 1H, J=3.4Hz, CH3-C-6). Ring II: δ=4 .88-4.87 (m, 1H, H-6), δ3.54-3.44 (m, 2H, H-1, H-4), 3.24 (dd, 1H, J = 13.4, 6.6 Hz, H-3), 3.15 (t, 1H, J = 7.7 Hz, H-5), 2.34-2.24 (m, 1H, H-2eq), 1.51-1.43 (m, 1H, H-2ax). The additional peaks in the spectrum are identified as follows: δ = 3.01 (s, 3H, NHSO2-CH3).

[0887] 13 C NMR (126MHz, MeOD): δ=95.73(C-1′), 77.24(C-4), 75.50(C-1), 73.70(C-5), 72.24(C-5′), 71.30(C-4′), 69.43(C -3′), 66.37(C-6′), 61.74(C-2′), 58.29(C-6), 51.86(C-3), 38.70(NHSO2-CH3), 33.14(C-2), 15.06(CH3-C-6′).

[0888] MALDI TOFMS: C 14 H 25 N7O9S([M+H] + )m / e 467.47; measured value is m / e 668.29.

[0889] 3′,4′,6′,6-Tetra-O-acetate-2′,3-diazide-1-N-benzamide-6′-(R)-methyl-paromomycinamine (Compound 5; Figure 4 ) Preparation:

[0890]

[0891] Compound 3 (0.380 g, 0.770 mmol) was dissolved in anhydrous pyridine (5 ml), the solution was cooled at -18 °C, and then acetic anhydride (4.5 eq., 0.3 ml, 3 mmol) was added. The reaction temperature was maintained at -18 °C, and the progress of the reaction was monitored by TLC (EtOAc / hexane 7:3), indicating that the reaction was complete after 12 hours. The reaction mixture was diluted with EtOAc (15 ml) and extracted with HCl 1N aqueous solution, saturated NaHCO3 aqueous solution, and brine. The combined organic layers were dried over MgSO4 and concentrated. The crude product was purified by silica gel chromatography (EtOAc / hexane 1:1) to give compound 5 (0.387 g, 76% yield).

[0892] 1 H NMR (500 MHz, CDCl3): Ring I: δ = 5.55-5.49 (m, 1H, H-3), 5.37 (d, 1H, J = 3.5 Hz, 4H, H-1), 5.02-4.96 (m, 2H, H-4, H-6), 4.35 (dd, 1H, J = 10.4, 1.8 Hz, H-5), 3.66-3.62 (m, 1H, H-2), 1.26 (d, 3H, J = 6.0 Hz, CH3-C-6). Ring II: δ = 6.66 (d, 1H, J = 7.4 Hz, amide), 4.92-4.87 (m, 1H, H-6), 4.27-4.20 (m, 1H, H-1), 3.84 (t, 1 H, J = 9.2 Hz, H-5), 3.55-3.47 (m, 1H, H-3), 3.46-3.39 (m, 1H, H-4), 2.67 (dt, 1H, J = 12.4, 4.2 Hz, H-2eq), 1.55-1.47 (m, 1H, H-2ax). Additional peaks in the spectrum were identified as follows: δ = 7.73-7.70 (m, 2H, Ph), 7.53 (dd, 1H, J = 4.8, 3.7 Hz, Ph), 7.43 (t, 2H, J = 7.6 Hz, Ph), 2.11 (s, 3H, Ac), 2.10 (s, 3H, Ac), 2.09 (s, 3H, Ac), 2.06 (s, 3H, Ac).

[0893] 13C NMR (126 MHz, CDCl3): δ = 172.61 (carbonyl), 170.12 (carbonyl), 169.95 (carbonyl), 169.94 (carbonyl), 167.06 (amide), 133.37 (Ph), 131.91 (Ph), 128.68 (Ph), 126.86 (Ph), 98.64 (C-1′), 83.50 (C-4), 74.67 (C-6), 74. 07(C-5), 71.34(C-3′), 70.90(C-5′), 69.08(C-4′), 68.85(C-6′), 61.64(C-2′), 58.41(C-3) , 48.65(C-1), 32.89(C-2), 21.01(Ac), 20.91(Ac), 20.69(Ac), 20.62(Ac), 14.01(CH3-C-6′).

[0894] MALDI TOFMS: C 23 H 33 N7O 12 ([M+H] + )m / e 661.1; measured value is m / e 622.1.

[0895] 3′,4′,6′,6-Tetra-O-acetate-2′,3-diazide-1-N-methanesulfonamide-6′-(R)-methyl-paromomycinamine (Compound 6; Figure 4 ) Preparation:

[0896]

[0897] Compound 6 was prepared as described for the preparation of compound 5 using compound 4 (0.8 g, 1.711 mmol) as starting material, and pyridine (10 ml) and acetic anhydride (5 eq, 0.8 ml, 9 mmol) to give 0.576 g (53%).

[0898] 1H NMR (500MHz, MeOD): Ring I: δ=5.89 (d, 1H, J=3.6Hz, H-1), 5.49-5.44 (m, 1H, H-3), 4.99 (dd, 2H, J=10.5, 9.0Hz, H-4, H-6), 4.44 (dd , 1H, J=10.6, 2.2Hz, H-5), 3.43-3.39 (m, 1H, H-2), 1.29 (d, 3H, J=5.9Hz, CH3-C-6). Ring II: δ=4.76-4.71 (m, 1H, H-6), 3.77 (t, 1H, J =9.5 Hz, H-5), 3.68 (ddd, 1H, J = 11.9, 10.0, 4.9 Hz, H-1), 3.57-3.48 (m, 2H, H-3, H-4), 2.39 (dt, 1H, J = 12.5, 4.5 Hz, H-2eq), 1.60 (dd, 1H, J = 25.7, 12.5 Hz, H-2ax). Additional peaks in the spectrum were identified as follows: δ = 2.97 (s, 3H, NHSO2-CH3), 2.13 (s, 3H, Ac), 2.07 (s, 6H, Ac), 2.06 (s, 3H, Ac).

[0899] 13 C NMR (126 MHz, MeOD): δ = 171.01 (carbonyl), 170.62 (carbonyl), 170.18 (carbonyl), 170.10 (carbonyl), 97.53 (C-1′), 78.80 (C-4), 75.69 (C-6), 74.53 (C-5), 70.05 (C-3′), 70.00 (C-5′), 69.34(C-4′), 68.66(C-6′), 60.54(C-2′), 59.47(C-1), 50.91(C-3), 40.42(NH SO2-CH3), 19.74(C-2), 19.70(Ac), 19.18(Ac), 19.15(Ac), 12.33(CH3-C-6′).

[0900] MALDI TOFMS: C 22 H 33 N7O 13 S([M+Na] + )m / e 635.60; measured value is m / e 658.05.

[0901] 3′,4′,6′,6-Tetra-O-acetate-2′,3-diazide-1-N-acetamide-6′-(R)-methyl-paromomycinamine (Compound 7; Figure 4 ) Preparation:

[0902]

[0903] Compound 7 was prepared as described for the preparation of compound 3 using compound 2 (0.1 g, 0.526 mmol) as starting material, and pyridine (3 ml) and acetic anhydride (6.4 eq, 0.2 ml, 1.6 mmol) to give 0.1 g (66%).

[0904] 1 H NMR (500MHz, MeOD): Ring I: δ=5.90 (d, 1H, J=3.5Hz, H-1), 5.50-5.44 (m, 1H, H-3), 5.02-4.96 (m, 2H, H-4, H-6), 4.45 (dd, 1H , J=10.5, 1.7Hz, H-5), 3.43-3.38 (m, 1H, H-2) 1.29 (d, 3H, J=5.2Hz, CH3-C-6). Ring II: δ=4.76 (t, 1H, J=10.1Hz, H-4), 4.09- 4.00 (m, 1H, H-1), 3.80 (t, 1H, J = 9.4 Hz, H-5), 3.71-3.62 (m, 1H, H-3), 3.55 (t, 1H, J = 9.7 Hz, H-6), 2.24-2.17 (m, 1H, H-2eq), 1.58 (q, 1H, J = 12.7 Hz, H-2ax). Additional peaks in the spectrum were identified as follows: δ = 2.08 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.06 (s, 3H, Ac), 1.90 (s, 3H, Ac).

[0905] 13 C NMR (126 MHz, MeOD): δ = 171.43 (amide), 171.07 (carbonyl), 170.57 (carbonyl), 170.16 (carbonyl), 170.10 (carbonyl), 97.55 (C-1′) 78.90 (C-3), 76.19 (C-6), 74.47 (C-1), 70.04 (C-3′), 70. 00(C-5′), 69.35(C-4′), 68.67(C-6′), 60.53(C-2′), 59.68(C-5), 32.23(C-2) , 21.21(Ac), 19.73(Ac), 19.39(Ac), 19.18(Ac), 19.16(Ac), 12.35(CH3-C-6′);

[0906] MALDI TOFMS: C 23 H 33 N7O 12 ([M+Na] + )m / e 599.55; measured value is m / e 622.08.

[0907] 3′,4′,6′,6-Tetra-O-acetate-2′,3-diazide-1-N-[(tert-butyloxy)carbonyl]-6′-(R)-methyl-paromomycinamine (Compound 8; Figure 4 ) Preparation:

[0908]

[0909] Compound 8 was prepared as described for the preparation of compound 5 using compound 1 (0.3 g, 0.610 mmol) as starting material, and pyridine (5 ml) and acetic anhydride (4.5 eq, 0.3 ml, 3 mmol) to give 0.060 g (15%).

[0910] 1 H NMR (400MHz, CDCl3): δ=5.56-5.45 (m, 1H), 5.35 (d, 1H, J=3.2Hz, H-1′), 4.97 (dd, J=13.2, 6 .4Hz, 2H), 4.71 (dd, J=19.8, 9.5Hz, 2H), 4.33 (dd, J=10.5, 1.6Hz, 1H), 3.79-3.67 (m, 2H), 3. 62(dd, J=10.5, 3.4Hz, 2H), 3.46-3.33(m, 2H), 2.49-2.38(m, 1H), 2.12(s, 3H, Ac), 2.09(s, 3 H, Ac), 2.07 (s, 3H, Ac), 2.05 (s, 3H, Ac), 1.42 (s, 11H, Boc), 1.25 (d, 3H, J=6.6Hz, CH3-C-6).

[0911] 13 C NMR (101MHz, CDCl3): δ = 171.58 (COOAc), 170.19 (COOAc), 170.00 (COOAc), 169.99 (COOAc) 155.22 (NHCOOC (CH3)3), 98.66(C-1′), 84.17, 83.74, 80.15, 74.54, 74.44, 71.44, 70.92, 69.15, 68.87, 61.74, 58.62, 48.99, 33.37, 28.27.

[0912] 3′,4′,6′,6-Tetra-O-acetate-2′,3-diazide-1-N-phenylsulfonamide-6′-(R)-methyl-paromomamide (Compound 9; Figure 4 ) Preparation:

[0913]

[0914] Compound 8 (0.460 grams, 0.7 mmol) was dissolved in dichloromethane (5 ml), and trifluoroacetic acid (1.5 ml) was added at ambient temperature. The progress of the reaction was monitored by TLC (EtOAc / hexane 1: 1), indicating that the reaction was complete after 1.5 hours. The reaction mixture was concentrated to dryness under reduced pressure to obtain 385 mg of N1-free product. The crude product was dissolved in chloroform (5 ml), and N, N-diisopropylethylamine (2.5 equivalents, 0.25 ml) and benzenesulfonyl chloride (1.5 equivalents, 0.1 ml) were then added. The progress of the reaction was monitored by TLC (EtOAc / hexane 3: 7), indicating that the reaction was complete after 24 hours. The reaction mixture was then concentrated to dryness under reduced pressure and purified by silica gel chromatography to obtain compound 9 (135 mg, 28%).

[0915] 1 H NMR (500MHz, CDCl3): Ring I: δ=5.46 (dd, 1H, J=10.4, 9.6Hz, H-3), 5.37 (d, 1H, J=3.5Hz, H-1), 4.98-4.93 (m, 2H, H-4, H-6), 4.30 (dd, 1H, J=10.5, 1. 8Hz, H-5), 3.56-3.52 (m, 1H, H-2), 1.22 (d, 3H, J=6.0Hz, CH3-C-6). Ring II: δ=5.53 (dd, 1H, J=8.2, 0.6Hz, H-4), 4.71 (t, 1H, J=10.0Hz, H-6), 3.67- 3.60 (m, 1H, H-5), 3.43-3.34 (m, 1H, H-1), 3.34-3.26 (m, 1H, H-3), 2.27 (dt, J = 12.6, 4.1 Hz, 1H, H-2eq), 1.58 (m, 1H, H-2ax). Additional peaks in the spectrum were identified as follows: δ = 7.86-7.83 (m, 2H, Ph), 7.62-7.58 (m, 1H, Ph), 7.53 (dd, 2H, J = 10.4, 4.7 Hz, Ph), 2.07 (s, 3H, Ac), 2.06 (s, 3H, Ac), 2.03 (s, 3H, Ac), 1.81 (s, 3H, Ac).

[0916] 13C NMR (126 MHz, CDCl3): δ = 172.01 (carbonyl), 170.19 (carbonyl), 170.01 (carbonyl), 169.94 (carbonyl), 140.88 (Ph 4°), 132.83(Ph), 129.31(Ph), 126.69(Ph), 98.50(C-1′), 82.60(C-4), 74.23(C-6), 74.13(C-5), 71.16(C-3′), 70.82(C-5′), 69.01(C-6 ′), 68.72(C-4′), 61.51(C-2′), 58.29(C-3), 52.05(C-1), 34.29(C-2), 21.01(Ac), 20.69(Ac), 20.66(Ac), 20.63(Ac), 13.87(CH3-C-6′).

[0917] MALDI TOFMS: C 27 H 35 N7O 13 S([M+Na] + )m / e 697.67; measured value is, m / e 720.048.

[0918] 5-O-(5″-azido-2″,3″-O-dibenzoyl-5″-deoxy-β-D-ribofuranose)-3′,4′,6′,6-tetra-O-acetate-2′,3-diazide-1-N-phenylsulfonamide-6′-(R)-methyl-paromomamide (Compound 12; Figure 5 ) Preparation:

[0919]

[0920] Flame dried powder Freshly distilled CH2Cl2 (5 ml) was added over 1% zeolite, followed by compound 9 (358 mg, 0.513 mmol) and donor compound 10 (1 gram, 2.05 mmol) prepared as described in Fridman, M. et al., Angew. Chemie, Int. Ed. 44, 447-452 (2005). The mixture was stirred at room temperature for 10 minutes and then cooled to -30°C. A catalytic amount of BF3.Et2O was added and the mixture was allowed to reach room temperature with stirring. The propagation of the reaction was monitored by TLC (EtOAc / hexane 3:7), which indicated completion after 2 hours. The reaction mixture was then diluted with EtOAc and filtered through Filter. Wash thoroughly with EtOAc After that, the washings were combined and concentrated. The crude product was purified by flash chromatography to give compound 12 (314 mg, 57%).

[0921] 1 H NMR (500MHz, CDCl3): Ring I: δ=5.90 (d, 1H, J=3.8Hz, H-1), 5.42 (dd, 1H, J=10.9, 9.2Hz, H-3), 5.00-4.93 (m, 1H, H-4), 4.43 (dd, 1H, J=10.5, 1.8Hz, H-5), 3.44-3.38 (m, 1H, H-2), 1.24 (d, 3H, J=5.9Hz, CH3-C-6 ). Ring II: δ=5.24 (d, 1H, J=8.7Hz, R1NHSO2R2), 4.81 (dd, 1H, J=9.9, 9.3Hz, H-5), 3.90-3.86 (m, 1H, H-4) , 3.72-3.67 (m, 1H, H-6), 3.52-3.39 (m, 2H, H-1, H-3), 2.33 (dt, 1H, J=11.8, 3.8Hz, H-2eq), 1.56-1.46 (m, 1H, H-2ax). Ring III: δ = 5.59 (d, 1H, J = 1.5 Hz, H-1), 5.49 (dd, 1H, J = 5.0, 1.1 Hz, H-2), 5.41 (dd, 1H, J = 7.1, 4.7 Hz, H-3), 4.48-4.44 (m, 1H, H-4), 3.61-3.58 (m, 1H, H-5). The additional peaks in the spectrum are identified as follows: δ = 7.93-7.90 (m, 2H, P h), 7.86-7.81 (m, 4H, Ph), 7.59 (dd, 2H, J=10.6, 4.3Hz, Ph), 7.55-7.49 (m, 3H, Ph), 7.41 (t, 2H, J=7.8 Hz, Ph), 7.33 (t, 2H, J=7.8Hz, Ph), 2.08 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.04 (s, 3H, Ac), 1.82 (s, 3H, Ac).

[0922] 13C NMR (126 MHz, CDCl3): δ = 171.62 (carbonyl), 170.15 (carbonyl), 170.00 (carbonyl), 169.92 (carbonyl), 165.24 (carbonyl), 165.16 (carbonyl), 140.84 (Ph 4°), 133.75(Ph), 133.63(Ph), 132.92(Ph), 129.68(Ph), 129.63(Ph), 129.37(Ph), 128.68(Ph), 128.57(Ph), 128 .50(Ph), 128.43(Ph), 126.68(Ph), 107.22(C-1″), 96.26(C-1′), 80.24(C-4), 80.20(C-4″), 76.96(C-6), 74.62( C-2″), 73.48(C-5), 71.62(C-3″), 70.53(C-3′), 70.24(C-5′), 68.94(C-6′), 68.53(C-4′), 61.38(C-2′), 58.73( C-1), 52.78(C-5″), 52.55(C-3), 34.68(C-2), 21.17(Ac), 20.72(Ac), 20.67(Ac), 20.57(Ac), 13.44(CH3-C-6′).

[0923] MALDI TOFMS: C 46 H 50 N 10 O 18 S([M+Na] + )m / e 1063.01; measured value is m / e 1085.13.

[0924] 5-O-(5″-azido-2″,3″-O-dibenzoyl-5″-deoxy-β-D-ribofuranose)-3′,4′,6′,6-tetra-O-acetate-2′,3-diazide-1-N-acetamide-6′-(R)-methyl-paromomycinamine (Compound 13; Figure 5 ) Preparation:

[0925]

[0926] Compound 13 was prepared as described for the preparation of compound 12 using compound 7 as starting material (398 mg, 0.663 mmol), and CH2Cl2 (10 ml), and donor 10 (1 gram, 1.722 mmol) to give 485 mg (75%).

[0927] 1 H NMR (500MHz, CDCl3): Ring I: δ=5.94 (d, 1H, J=3.8Hz, H-1), 5.47-5.41 (m, 1H, H-3), 5.02-4.94 (m, 2H, H-5, H-6), 4.48 (dd, 1H, J=10.6, 1.8Hz, H-4), 3.47 -3.41 (m, 1H, H-2), 1.26 (d, 3H, J=6.2Hz, CH3-C-6). Ring II: δ=5.79 (d, 1H, J=8.0Hz, RNHCO), 4.89-4.84 (m, 1H, H-4), 4.09 (ddd, 1H, J=9.5, 8.3, 5.2Hz, H-1), 3.98 (t, 1H, J=9.1Hz, H-5), 3.75-3.69 (m, 1H, H-6), 3.64-3.55 (m, 1 H, H-3), 2.47 (dt, 1H, J=8.7, 4.6Hz, H-2eq), 1.38 (t, 1H, J=12.8Hz, H-2ax ). Ring III: δ = 5.64 (d, 1H, J = 1.5Hz, H-1), 5.60 (d, 1H, J = 5.9Hz, H-2), 5.48 (d d, 1H, J=7.3, 4.8Hz, H-3), 4.52-4.48 (m, 1H, H-4), 3.64-3.60 (m, 2H, H-5).

[0928] 13 C NMR (126 MHz, CDCl3): δ = 172.05 (amide), 170.09 (carbonyl), 170.02 (carbonyl), 169.88 (carbonyl), 169.79 (carbonyl), 165.24 (carbonyl), 165.21 (carbonyl), 133.72 (Ph), 133.64 (Ph), 129.72 (Ph), 129.66 (Ph), 128.70 (Ph), 128.56 (Ph), 128.53 (Ph), 128.44 (Ph), 107.46 (C-1″), 96.30 (C-1′), 80.44 (C-5), 80.15 (C-4″), 77.20(C-6), 74.65(C-2″), 74.06(C-4), 71.56(C-3″), 70.62(C-4′), 70.21(C-3′), 68.98(C-5′), 68.62(C-6′), 61.44(C-2′), 58.84(C-3), 52.75(C-5″), 48.24(C-4), 32.94(C-2), 23.18(RNHAc), 21.19(Ac), 20.88(Ac), 20.75(Ac), 20.68(Ac), 13.58(CH3-C-6′).

[0929] MALDI TOFMS: C 42 H 48 N 10 O 17 ([M+Na] + )m / e 964.89; measured value is m / e 987.15.

[0930] 5-O-(5″-azido-2″,3″-O-dibenzoyl-5″-deoxy-β-D-ribofuranose)-3′,4′,6′,6-tetra-O-acetate-2′,3-diazide-1-N-methanesulfonamide-6′-(R)-methyl-paromomycinamine (Compound 14; Figure 5 ):

[0931]

[0932] Compound 14 was prepared as described for the preparation of compound 12 using compound 6 as starting material (0.680 g, 0.663 mmol), as well as CH2Cl2 (10 ml), and donor 10 (2 g, 4 mmol) to give 0.73 g (75%).

[0933] 1H NMR (500MHz, CDCl3): Ring I: δ=5.91 (d, 1H, J=3.7Hz, H-1), 5.46-5.40 (m, 1H, H-3), 5.01-4.94 (m, 2H, H-4, H- 6), 4.44 (dd, 1H, J=10.5, 1.6Hz, H-5), 3.45 (dd, 1H, J=10.7, 4.1Hz, H-2), 1.25 (d, 1H, J=5.7Hz, CH3-C-6) . Ring II: δ=5.06 (d, 1H, J=9.5Hz, NHSO2-CH3), 4.92 (t, 1H, J=9.7Hz, H-6), 3.95 (t, 1H, J=8.9Hz, H-5), 3.79- 3.73 (m, 1H, H-4), 3.61-3.49 (m, 2H, H-1, H-3), 2.47 (dt, 1H, J=12.3, 4.0Hz, H-2), 1.60 (dd, 1H, J=26.6, 1 2.0 Hz, H-2). Ring III: δ = 5.64 (d, 1H, J = 0.9 Hz, H-1), 5.59 (d, 1H, J = 5.7 Hz, H-2), 5.45 (dd, 1H, J = 7.1, 4.7 Hz, H-3), 4.51 (d, 1H, J = 7.0 Hz, H-4), 3.62 (d, 2H, J = 4.5 Hz, H-5, H-5). Additional peaks in the spectrum were identified as follows: δ = 7.93 (d, 2H, J = 7.4 Hz, Ar), 7.86 (d, 2H, J=7.4Hz, Ar), 7.55 (dt, 2H, J=23.6, 7.5Hz, Ar), 7.41 (t, 2H, J=7.8Hz, Ar), 7.34 (t, 2H, J =7.8Hz, Ar), 2.97 (s, 3H, NHSO2-CH3), 2.25 (s, 2H, Ac), 2.08 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.05 (s, 3H, Ac).

[0934] 13C NMR (126 MHz, CDCl3): δ = 171.45 (carbonyl), 170.12 (carbonyl), 169.97 (carbonyl), 169.86 (carbonyl), 165.33 (carbonyl), 165.19 (carbonyl), 133.73 (Ar), 133.63 (Ar), 129.70 (Ar), 129.65 (Ar), 128.71 (Ar), 128.56 (Ar), 128.54 (Ar), 128.45 (Ar), 107.40 (C-1″), 96.28 (C-1′), 80.39 (C-4), 80.05 (C-6′), 7 7.09(C-6), 74.66(C-3), 73.47(C-2″), 71.71(C-5′), 70.61(C-4′), 70.30(C-3′), 68.99(C-3″), 68.56(C-4″), 61.46(C-2′), 58.84( C-1), 52.83(C-3), 52.14(C-5″), 42.08(NHSO2-CH3), 34.82(C-2), 21.15(Ac), 21.06(Ac), 20.70(Ac), 20.64(Ac), 13.50(CH3-C-6′).

[0935] MALDI TOFMS: C 41 H 48 N 10 O 18 S([M+Na] + )m / e 1000.94; measured value is m / e 1023.31.

[0936] 5-O-(5″-azido-2″,3″-O-dibenzoyl-5″-deoxy-β-D-ribofuranose)-3′,4′,6′,6-tetra-O-acetate-2′,3-diazide-1-N-dibenzamide-6′-(R)-methyl-paromomycinamine (Compound 15; Figure 5 ) Preparation:

[0937]

[0938] Compound 15 was prepared as described for the preparation of compound 12 using compound 5 as starting material (0.35 g, 0.529 mmol), anhydrous CH2Cl2 (10 ml), and donor 10 (0.864 g, 2.11 mmol) to give 0.4 g (73%).

[0939] 1H NMR (500MHz, CDCl3): Ring I: δ=5.98 (d, 1H, J=3.6Hz, H-1), 5.49-5.43 (m, 1H, H-3), 5.04-4.96 (m, 2H, H-4, H-6), 4.51 (d, 1H, J=9.8Hz, H-5), 3.48-3.43 (m, 1H, H-2), 1.27 (d, 1H, J=5.7Hz, CH3-C-6). Ring II: δ=6.59 (d, 1H, J=7.6Hz, amide), 5.02 (t, 1H, J=9.8Hz, H-6), 4.31-4.24 (m, 1H, H-1), 4.06 (t, 1H, J=8.9Hz, H-5), 3 .81-3.73 (m, 1H, H-4), 3.66 (tt, 1H.J=9.7, 4.8Hz, H-3), 2.64 (dd, 1H, J=10.1, 3.1Hz, H-2eq), 1.51-1.4 2 (m, 1H, H-2ax). Ring III: δ = 5.69 (s, 1H, H-1), 5.64 (d, 1H, J = 4.7 Hz, H-2), 5.51 (dd, 1H, J = 6.5, 4.8 Hz, 1H), 4.51 (d, 1H, J = 7.4 Hz, H-3), 3.64 (m, 2H, H-5). Additional peaks in the spectrum were identified as follows: δ = 7.94 (d, 2H, J = 7.6 Hz, Ph), 7.85 (d, 2H, J = 7.7 Hz, Ph). =7.7Hz, Ph), 7.71 (d, 2H, J = 7.5Hz, Ph), 7.58 (t, 1H, J = 7.3Hz, Ph), 7.55-7.49 (m, 2H, Ph), 7.47-7.39 (m , 4H, Ph), 7.33 (t, 2H, J=7.7Hz, Ph), 2.15 (s, 3H, Ac), 2.09 (s, 3H, Ac), 2.08 (s, 3H, Ac), 2.06 (s, 3H, Ac).

[0940] 13C NMR (126 MHz, CDCl3): δ = 172.55 (amide), 170.08 (carbonyl), 170.01 (carbonyl), 169.85 (carbonyl), 166.80 (carbonyl), 165.22 (carbonyl), 165.21 (carbonyl), 133.69 (Ph), 133.61 (Ph), 133.21 (Ph), 131.96 (Ph), 129.71 (Ph), 129.65 (Ph), 128.74 (Ph), 128.55 (Ph), 128.42 (Ph), 126.83 (Ph), 107.55 (C-1″), 96.37 (C-1′) , 80.64(C-5), 80.13(C-4″), 77.32(C-4), 74.69(C-2″), 74.23(C-6), 71.56(C-3″), 70.66(C-5′), 70.25(C-3′), 69.03(C-6′), 68.63(C-4 ′), 61.50(C-2′), 58.94(C-3), 52.77(C-5″), 48.97(C-1), 33.04(C-2), 21.17(Ac), 20.92(Ac), 20.73(Ac), 20.66(Ac), 13.58(CH3-C-6′).

[0941] MALDI TOFMS: C 47 H 50 N 10 O 17 ([M+H] + )m / e 1026.34; measured value is m / e 1027.28.

[0942] 5-O-(5″-(S)-azido-2″,3″-O-dibenzoyl-5″-deoxy-β-D-ribofuranose)-3′,4′,6′,6-tetra-O-acetate-2′,3-diazide-1-N-methanesulfonamide-6′-(R)-methyl-paromomamide (Compound 16; Figure 5 ) Preparation:

[0943]

[0944] Compound 16 was prepared as described for the preparation of compound 12 using compound 6 (0.4 g, 0.629 mmol) as starting material, anhydrous CH2Cl2 (15 ml) and donor compound 11 (1.4 g, 2.5 mmol) prepared as described in Kandasamy, J. et al., Medchemcomm 2, 165-171 (2011), yielding 0.37 g (58%).

[0945] 1 H NMR (500MHz, CDCl3): Ring I: δ=5.92 (d, 1H, J=3.8Hz, H-1), 5.44-5.37 (m, 1H, H-3), 4.97 (dd, 2H, J=17.2, 7. 6Hz, H-4, H-6), 4.43 (d, 1H, J=10.3Hz, H-5), 3.53 (dd, 1H, J=10.8, 3.9Hz, H-2), 1.24 (d, 3H, J=5.6Hz, CH3 -C-6). Ring II: δ=5.11 (d, 1H, J=9.4Hz, NHSO2-CH3), 4.92 (t, 1H, J=9.6Hz, H-5), 3.92 (t, 1H, J=8.9Hz, H-6), 3.76 (t, 1H, J=9.1Hz, H-4), 3.55 (ddd, 2H, J=17.4, 10.2, 5.3Hz, H-1, H-3), 2.48 (dd, 1H, J=8.1, 4.0Hz, H- 2eq), 1.61 (dd, 1H, J=26.0, 12.3Hz, H-2ax). Ring III: δ=5.61 (d, 2H, J=4.5Hz, H-1, H-2), 5.48 (dd, 1H, J=7.8 , 4.7Hz, H-3), 4.31 (t, 1H, J=6.8Hz, H-4), 3.72 (p, 1H, J=6.9Hz, H-5), 1.28 (d, 3H, J=7.0Hz, CH3-C-5′).Fig. Additional peaks in the spectrum were identified as follows: δ = 7.89 (dd, 2H, J = 27.3, 7.7 Hz, Ar), 7.55 (dt, 2H, J = 20.8, 7.4 Hz, Ar), 7.37 (dt, 2H, J = 30.8, 7.7 Hz, Ar), 2.98 (s, 3H, NHSO2-CH3), 2.32 (s, 3H, Ac), 2.08 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.05 (s, 3H, Ac).

[0946] 13C NMR (126 MHz, CDCl3): δ = 171.65 (carbonyl), 170.17 (carbonyl), 170.03 (carbonyl), 169.86 (carbonyl), 165.35 (carbonyl), 165.01 (Ar), 133.73 (Ar), 133.64 (Ar), 129.71 (Ar), 129.63 (Ar), 128.74 (Ar), 128.68 (Ar), 128.56 (Ar), 128.51 (Ar), 128.45 (Ar), 107.60 (C-1″), 96.14 (C-1′), 83.99 (C-4″), 79.92 (C-6), 77. 38(C-4), 74.72(C-2″), 73.15(C-5), 71.62(C-3″), 70.78(C-3′), 70.2 2(C-5′), 69.00(C-4′), 68.56(C-6′), 61.67(C-2′), 59.02(C-5″), 58. 72(C-1), 52.30(C-3), 42.04(NHSO2-CH3), 34.85(C-2), 21.17(Ac), 21.11(Ac), 20.72(Ac), 20.66(Ac), 15.49(CH3-C-5′), 13.48(CH3-C-6′).

[0947] MALDI TOFMS: C 42 H 50 N 10 O 18 S([M+Na] + )m / e 1014.97; measured value is m / e 1037.28.

[0948] 5-O-(5"-(S)-azido-2",3"-O-dibenzoyl-5"-deoxy-β-D-ribofuranose)-3',4',6',6-tetra-O-acetate-2',3-diazide-1-N-acetamide-6'-(R)-methyl-paromomycinamine (Compound 17; Figure 5 ) Preparation:

[0949]

[0950] Compound 17 was prepared as described for the preparation of compound 12 using compound 7 as starting material (1.5 g, 2.5 mmol), anhydrous CH2Cl2 (15 ml), and donor 11 (4.5 g, 10 mmol) to afford 1.6 g (64%).

[0951] 1H NMR (500MHz, CDCl3): Ring I: δ=5.95 (d, 1H, J=3.6Hz, H-1), 5.41 (dd, 1H, J=10.6, 8.8Hz, H-3), 5.01-4.93 (m, 2H, H-6, H-4), 4.46 (dd, 1H, J=10.6, 1.3Hz, H-5), 3.50 (dd, 1H, J=10.6, 3.8Hz, H-2), 1.25 (d, 3H, J= 6.0Hz, CH3-C-6). Ring II: δ=5.93 (d, 1H, J=8.3Hz, NH-Ac), 4.87 (t, 1H, J=9.8Hz, H-6), 4.14-4.04 (m, 1H, H-1), 3.94 (t, 1H, J=8.9Hz, H-5), 3.79-3.68 (m, 1H, H-4), 3.60 (dt, 1H, J=11.6, 5.2Hz, H-3), 2.48-2.4 1(m, 1H, H-2eq), 1.39 (dd, 1H, J=26.8, 13.1Hz, H-2ax). Ring III: δ=5.61 (d, 2H, J=4.4Hz, H-1, H-2), 5.51 (dd, 1H, J=7.9, 4.5Hz, H-3), 4.31-4.27 (m, 1H, H-4), 3.70 (m, 1H, H-5), 1.29 (d, 3H, J=6.9Hz, CH3-C-5′ Additional peaks in the spectrum were identified as follows: δ = 7.88 (dd, 4H, J = 33.7, 7.4 Hz, Ar), 7.54 (dt, 2H, J = 24.3, 7.4 Hz, Ar), 7.36 (dt, 4H, J = 36.9, 7.8 Hz, Ar), 2.22 (s, 3H, Ac), 2.08 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.04 (s, 3H, Ac), 1.94 (s, 3H, Ac).

[0952] 13C NMR (126 MHz, CDCl3): δ = 172.16 (carbonyl), 170.09 (carbonyl), 170.01 (carbonyl), 169.82 (carbonyl), 169.7 (carbonyl), 165.23 (carbonyl), 165.04 (carbonyl), 133.68 (Ar), 133.60 (Ar), 129.71 (Ar), 129.67 (Ar), 129.63 (Ar), 128.69 (Ar), 128.54 (Ar), 128.42 (Ar), 107.60 (C-1″), 96.13 (C-1′), 83.64 (C-4″), 80.41 ( C-5), 77.54(C-4), 74.78(C-2″), 73.82(C-6), 71.47(C-3″), 70.82(C-5′), 70.17(C-3′), 69.03(C-6′), 68.63(C-4′), 61.64(C-2′), 58 .82(C-5″), 48.23(C-1), 32.80(C-2), 23.15(Ac), 21.14(Ac), 20.89(Ac), 20.71(Ac), 20.65(Ac), 15.51(CH3-C-5″), 13.57(CH3-C-6′).

[0953] MALDI TOFMS: C 43 H 50 N 10 O 17 ([M+Na] + )m / e 978.91; measured value is m / e 1001.32.

[0954] 5-O-(5″-(S)-azido-2″,3″-O-dibenzoyl-5″-deoxy-β-D-ribofuranose)-3′,4′,6′,6-tetra-O-acetate-2′,3-diazide-1-N-benzamide-6′-(R)-methyl-paromomycinamine (Compound 18; Figure 5 ) Preparation:

[0955]

[0956] Compound 18 was prepared as described for the preparation of compound 12 using compound 5 as starting material (0.323 g, 0.488 mmol), anhydrous CH2Cl2 (15 ml), and donor 11 (1.4 g, 1 mmol) to give 0.422 g (83%).

[0957] 1H NMR (500MHz, CDCl3): Ring I: δ=5.99 (d, 1H, J=3.8Hz, H-1), 5.44 (t, 1H, J=10.3Hz, H-3), 5.00 (dd, 2H, J=19.0, 8.7Hz , H-5, H-6), 4.49 (dd, 1H, J=10.3, 1.3Hz, H-4), 3.53 (dd, 1H, J=10.6, 4.1Hz, H-2), 1.26 (d, 3H, J=5.8Hz, CH3-C-6) . Ring II: δ=6.62 (d, 1H, J=7.8Hz, NHCOBz), 5.03 (d, 1H, J=10.1Hz, H-6), 4.33-4.23 (m, 1H, H-1), 4.02 (t, 1H, J=8.9H z, H-5), 3.77 (t, 1H, J=9.2Hz, H-4), 3.69-3.62 (m, 1H, H-3), 2.64 (dt, 1H, J=7.9, 5.1Hz, H-2eq), 1.46 (q, 1H, J=12 .2 Hz, H-2ax). Ring III: δ = 5.67 (d, 2H, J = 4.5 Hz, H-1, H-2), 5.54 (dd, 1H, J = 7.8, 4.9 Hz, H-3), 4.34-4.27 (m, 1H, H-4), 3.72 (p, 1H, J = 7.0 Hz, H-5), 1.31 (d, 3H, J = 6.9 Hz, CH3-C-5′). Additional peaks in the spectrum were identified as follows: δ = 7.92 (d, 2H, J = 7.5 Hz, Ar), 7.84 (d , 2H, J=7.6Hz, Ar), 7.72 (d, 2H, J=7.4Hz, Ar), 7.57 (t, 1H, J=7.4Hz, Ar), 7.52 (t, 2H, J=7.4Hz, Ar), 7.43 (dt, 4H, J =19.0, 7.6Hz, Ar), 7.33 (t, 2H, J = 7.8Hz, Ar), 2.22 (s, 3H, Ac), 2.09 (s, 3H, Ac), 2.08 (s, 3H, Ac), 2.06 (s, 3H, Ac).

[0958] 13C NMR (126 MHz, CDCl3): δ = 172.81 (carbonyl), 170.10 (carbonyl), 170.03 (carbonyl), 169.82 (carbonyl), 166.79 (carbonyl), 165.21 (carbonyl), 165.02 (carbonyl), 133.66 (Ar), 133.58 (Ar), 133.25 (Ar), 131.95 (Ar), 129.72 (Ar), 129.63 (Ar), 128.74 (Ar), 128.53 (Ar), 128.41 (Ar), 126.84 (Ar), 107.74 (C-1″), 96.23 (C-1′), 83.67 (C-1′). -4″), 80.55(C-5), 77.64(C-4), 74.84(C-2″), 73.91(C-6), 71.46(C-3 ″), 70.85(C-4′), 70.20(C-4′), 69.05(C-6′), 68.66(C-5′), 61.70(C-2 ′), 58.90(C-5″), 58.88(C-3), 49.09(C-1), 32.96(C-2), 21.17(Ac), 20.97(Ac), 20.73(Ac), 20.66(Ac), 15.53(CH3-C-5′), 13.60(CH3-C-6′).

[0959] MALDI TOFMS: C 47 H 50 N 10 O 17 ([M+H2O] + )m / e 1046.98; measured value is m / e 1065.6.

[0960] 6′-(R)-methyl-5-O-(5″-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-phenylsulfonamide paromomamide (Compound 19; Figure 5 ) Preparation:

[0961]

[0962] Compound 12 (314 mg, 0.295 mmol) was treated with a solution of MeNH2 (33% solution in EtOH, 5 ml). The propagation of the reaction was monitored by TLC (EtOAc / MeOH, 7:3), indicating completion after 12 hours. The reaction mixture was then evaporated to dryness and purified by silica gel chromatography (100% EtOAc) to give compound 19 (0.148 g, 73%).

[0963] 1H NMR (500MHz, MeOD): Ring I: δ=6.00 (d, 1H, J=3.6Hz, H-1), 4.02 (dd, 1H, J=6.8, 3 .5Hz, H-5), 3.94-3.86 (m, 2H, H-3, H-6), 3.32 (dd, 1H, J=10.0, 8.7Hz, H-4), 3 .11 (dd, 1H, J=10.6, 4.3Hz, H-2), 1.21 (d, 1H, J=6.0Hz, CH3-C-6). Ring II: δ=3. 60 (m, 2H, J=17.3, 9.0Hz, H-4, H-5), 3.43-3.35 (m, 1H, H-1), 3.27-3.13 (m, 2H , H-3, H-6), 1.85 (dt, 1H, J = 12.3, 4.0 Hz, H-2eq), 1.33-1.20 (m, 1H, H-2ax). Ring III: δ = 5.29 (d, 1H, J = 0.8 Hz, H-1), 4.14 (d, 1H, J = 5.1 Hz, H-2), 4.02 (dd, 1H, J = 7.5, 4.1 Hz, H-3), 3.98 (td, 1H, J = 6.3, 2.9 Hz, H-4), 3.55 (dd, 1H, J = 13.3, 2.7 Hz, H-5), 3.46 (dd, 1H, J = 13.2, 6.2 Hz, H-5). The additional peaks in the spectrum were identified as follows: δ = 7.92 (dd, 2H J=5.2, 3.4Hz, Ph), 7.63 (ddd, 1H, J=8.6, 2.4, 1.2Hz, Ph), 7.60-7.54 (m, 2H, Ph).

[0964] 13 C NMR (126MHz, MeOD): δ = 142.82 (Ph 4°), 133.63(Ph), 130.16(Ph), 127.95(Ph), 111.21(C-1″), 97.10(C-1′), 85.77(C-5), 82.13(C-4″), 76.39(C-4), 76.24(s), 76.00(C-6), 75.01(C- 3′), 74.10(C-4′), 72.57(C-5′), 72.46(C-6′), 68.97(C-3″), 64.85(C-2′ ), 61.38(C-1), 54.82(C-3), 54.39(C-5″), 34.40(C-2), 17.65(CH3-C-6′).

[0965] 6′-(R)-methyl-5-O-(5″-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-acetamide paromycinamine (Compound 20; Figure 5) Preparation:

[0966]

[0967] Compound 20 was prepared as described for the preparation of compound 19 using compound 13 as starting material (485 mg, 0.824 mmol) and MeNH 2 (33% solution in EtOH, 10 ml) to give 278 mg (93%).

[0968] 1 H NMR (500MHz, MeOD): Ring I: δ=5.99 (d, 1H, J=3.6Hz, H-1), 4.00 (dd, 1H, J=6.8, 3.6Hz, H-6), 3 .93 (dd, 1H, J=9.9, 4.1Hz, H-4), 3.88 (dd, 1H, J=10.1, 9.1Hz, H-3), 3.28 (dd, 1H, J=10.1, 8 .7Hz, H-5), 3.07 (dd, 1H, J=10.6, 4.3Hz, H-2), 1.19 (d, 1H, J=6.0Hz, CH3-C-6). Ring II: δ=3. 75(ddd, 1H, J=12.0, 10.4, 4.5Hz, H-1), 3.69-3.63 (m, 1H, H-4), 3.60 (t, 1H, J=8.9Hz, H-5) , 3.49 (ddd, 1H, J=12.3, 9.7, 4.8Hz, H-3), 3.28 (dd, 1H, J=10.4, 8.9Hz, m, 1H, H-6), 2.09 ( dt, 1H, J=12.5, 4.3Hz, H-2eq), 1.32 (dd, 1H, J=25.7, 12.6Hz, H-2ax). Ring III: δ=5.29 (d, 1H, J=1.1Hz, H-1), 4.12 (d, 1H, J=5.3Hz, H-2), 3.99 (dd, 1H, J=7.5, 4.2Hz, H-3), 3.95 (dd, 1H, J=7.0, 3.4Hz, H-4), 3.52 (dd, 1H, J=13.2, 2.7Hz, H-5), 3.44 (dd, 1H, J=13.2, 6.1Hz, H-5).

[0969] 13C NMR (126MHz, MeOD): δ=173.48 (amide), 111.24 (C-1″), 97.25 (C-1′), 86.33 (C- 5), 82.27(C-4″), 76.66(C-2″), 76.36(C-5′), 76.14(C-4), 75.14(C-4′), 74 .20(C-3″), 72.67(C-6), 72.52(C-3′), 69.03(C-6′), 64.97(C-2′), 61.96(C -3), 54.48(C-5″), 50.71(C-1), 33.85(C-2), 22.80(Ac), 17.63(CH3-C-6′).

[0970] MALDI TOF MS: C 20 H 32 N 10 O 11 ([M+Na] + )m / e 588.53; measured value is m / e 611.11.

[0971] 6′-(R)-methyl-5-O-(5″-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-methanesulfonamide paromomamide (Compound 21; Figure 5 ) Preparation:

[0972]

[0973] Compound 21 was prepared as described for the preparation of compound 19 using compound 14 (0.73 g, 0.729 mmol) and MeNH2 (33% solution in EtOH, 10 ml) to give 0.426 g (93%).

[0974] 1H NMR (500MHz, MeOD): Ring I: δ=6.03 (d, 1H, J=3.2Hz, H-1), 4.06 (dd, 1H, J=7.0, 3.3Hz, H-6), 4.00-3.91 (m, 2H, H-5, H-3), 3.39-3.32 (m, 1H , H-4), 3.14 (dd, 1H, J=10.7, 4.8Hz, H-2), 1.26 (d, 3H, J=4.8Hz, CH3-C-6). Ring II: δ=3.74-3.64 (m, 2H, H-4, H-6), 3.60-3.52 (m, 1H, H-1) , 3.30 (dd, 1H, J = 9.5, 4.6 Hz, H-3), 2.28 (dd, 1H, J = 8.4, 2.9 Hz, H-2eq), 1.46 (dd, 1H, J = 26.2, 12.4 Hz, H-1ax). Ring III: δ = 5.37 (s, 1H, H-1), 4.19 (d, 1H, J = 4.2 Hz, H-2), 4.06 (t, 2H, J = 5.5 Hz, H-3, H-4), 3.62-3.48 (m, 2H, H-5, H-5), the additional peaks in the spectrum are identified as follows: δ = 3.06 (s, 3H, NHSO2-CH3).

[0975] 13 C NMR (126MHz, MeOD): δ=109.65(C-1″), 95.82(C-1′), 84.58(C-4), 80.93(C -6′), 75.09(C-6), 74.92(C-3), 74.89(C-2″), 73.70(C-5′), 72.81(C-4′), 71.20(C-3′), 71.10(C-3″), 67.68(C-4″), 63.52(C-2′), 60.33(C-1), 53.5 0(C-3), 53.03(C-5″), 40.26(NHSO2-CH3), 34.46(C-2), 16.31(CH3-C-6′).

[0976] MALDI TOF MS: ([MH] + )m / e 624.58; measured value is m / e 623.19.

[0977] 6′-(R)-methyl-5-O-(5″-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-benzamide paromomamide (Compound 22; Figure 5 ) Preparation:

[0978]

[0979] Compound 22 was prepared as described for the preparation of compound 19 using compound 15 (0.4 g, 0.389 mmol) and a solution of MeNH2 (33% solution in EtOH, 10 ml) to give 0.25 g (95%).

[0980] 1 H NMR (500MHz, MeOD): Ring I: δ=6.08 (d, 1H, J=3.3Hz, H-1), 4.07 (dd, 1H, J=7.1, 3.4Hz, H-6) , 4.03-3.95 (m, 2H, H-3, H-5), 3.42-3.37 (m, 1H, H-4), 3.16 (dd, 1H, J=10.6, 4.7Hz, H-2) , 1.28 (d, 1H, J = 4.8 Hz, CH3-C-6). Ring II: δ = 8.35 (d, 1H, J = 7.9 Hz, amide), 4.08 (td, 2H, J = 11.1, 4.8 Hz, H-1, H-5), 3.81-3.70 (m, 1H, H-4), 3.59 (dt, 2H, J = 18.7, 7.0 Hz, H-6, H-3), 2.2 5 (dd, 1H, J=8.0, 4.0Hz, H-2eq), 1.57 (dd, 1H, J=26.2, 12.8Hz, H-2ax). Ring III: δ=5.41 (s , 1H, H-1), 4.22 (d, 1H, J=5.2Hz, H-2), 4.09 (dd, 1H, J=7.8, 3.8Hz, H-3), 4.06-4.02 (m, 1 H, H-4), 3.59 (dd, 1H, J = 13.1, 2.3 Hz, H-5), 3.51 (dd, 1H, J = 13.1, 6.3 Hz, H-5). The additional peaks in the spectrum were identified as follows: δ = 7.89 (d, 2H, J = 7.4 Hz, Ph), 7.56 (t, 1H, J = 7.3 Hz, Ph), 7.48 (t, 2H, J = 7.6 Hz, Ph).

[0981] 13C NMR (126 MHz, MeOD): δ = 169.01 (amide), 134.16 (Ph), 131.39 (Ph), 128.11 (Ph), 127.06 (Ph), 109.87 (C-1″), 95.88 (C-1′), 84.94 (C-6), 80.91 (C-4″), 75.39 (s), 74.99 (C- -2″), 74.51(C-4), 73.71(C-5′), 72.94(C-4′), 71.25(C-3″), 71.11(C-3′), 67.86 (C-6″), 63.52(C-2′), 60.67(C-3), 53.10(C-5″), 32.30(C-2), 16.54(CH3-C-6′).

[0982] 6′-(R)-methyl-5-O-(5″-(S)-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-methanesulfonamide paromomamide (Compound 23; Figure 5 ) Preparation:

[0983]

[0984] Compound 23 was prepared as described for the preparation of compound 19 using compound 16 (0.37 g, 0.364 mmol) and a solution of MeNH2 (33% solution in EtOH, 10 ml) to give 0.230 g (98%).

[0985] 1H NMR (500MHz, MeOD): Ring I: δ=6.04 (d, 1H, J=3.6Hz, H-1), 4.02 (dd, 1H, J=6.8, 3. 3Hz, H-6), 3.95-3.89 (m, 2H, H-3, H-5), 3.29 (dd, 1H, J=10.0, 8.7Hz, H-4), 3.0 8 (dd, 1H, J=10.6, 4.4Hz, H-2), 1.21 (d, 3H, J=6.0Hz, CH3-C-6). Ring II: δ3.68 (t, 1H, J=9.6Hz, H-4), 3.61 (t, 1H, J=8.5Hz, H-5), 3.54-3.47 (m, 1H, H-1), 3.29-3 .20 (m, 2H, H-3, H-6), 2.27-2.19 (m, 1H, H-2eq), 1.42 (dd, 1H, J = 27.1, 11.5 Hz, H-2ax). Ring III: δ = 5.30 (d, 1H, J = 0.5 Hz, H-1), 4.14 (d, 1H, J = 5.4 Hz, H-2), 4.08 (dd, 1H, J = 7.7, 4.2 Hz, H-3), 3.72 (t, 1H, J = 6.4 Hz, H-4), 3.67-3.60 (m, 1H, H-5), 1.33 (d, 3H, J = 6.9 Hz, CH3-C-5′). The additional peaks in the spectrum were identified as follows: δ = 3.01 (s, 3H, NHSO2-CH3).

[0986] 13 C NMR (126MHz, MeOD): δ=109.27(C-1″), 95.81(C-1′), 84.65(C-4″), 84.53(C-5) , 75.12(C-2″), 74.94(C-4), 74.83(C-6), 73.73(C-3′), 72.77(C-4′), 71.27(C- 3″), 71.15(C-5′), 67.51(C-6″), 63.41(C-2′), 60.37(C-1), 59.27(C-5″), 53.5 0(C-3), 40.23(NHSO2-CH3), 34.47(C-2), 16.10(CH3-C-5′), 14.66(CH3-C-6′).

[0987] MALDI TOFMS: C 20 H 34 N 10 O 12 S([M+H] - )m / e 638.61; measured value is m / e 637.5.

[0988] 6′-(R)-methyl-5-O-(5″-(S)-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-acetamide paromycinamine (Compound 24; Figure 5 ) Preparation:

[0989]

[0990] Compound 24 was prepared as described for the preparation of compound 19 using compound 17 (1.6 g, 1.6 mmol) and a solution of MeNH2 (33% solution in EtOH, 10 ml) to give 0.820 g (81%).

[0991] 1 H NMR (500MHz, MeOD): Ring I: δ=6.07 (d, 1H, J=3.4Hz, H-1), 4.06-4.00 (m, 1H, H-6), 3.98-3.91 (m, 2H, H-3, H-5), 3.31 (dd, 1H, J=10.0, 8.8Hz, H-4), 3.09 (d d, 1H, J=10.6, 4.5Hz, H-2), 1.23 (d, 3H, J=5.0Hz, CH3-C-6). Ring II: δ=3.81-3 .74(m,1H,H-1),3.73-3.68(m,1H,H-4,),3.68-3.61(m,1H,H-3),3.56-3.4 8(m, 1H, H-3), 3.32 (dd, 1H, J=10.0, 9.4Hz, H-6), 2.12 (dt, 1H, J=12.1, 4.1 Hz, H-2eq), 1.35 (dd, 1H, J=25.9, 12.6Hz, H-2ax). Ring III: δ=5.30 (s, 1H, H-1 ), 4.14 (d, 1H, J = 5.3Hz, H-2), 4.11-4.06 (m, 1H, H-3), 3.73 (t, 1H, J = 5.8Hz , H-4), 3.66 (dd, 1H, J=11.5, 3.6Hz, H-5), 1.34 (d, 3H, J=6.7Hz, CH3-C-5′).

[0992] 13C NMR (126MHz, MeOD): δ=108.70(C-1″), 95.10(d, C-1′), 84.38(C-5), 83.97(C-4″ ), 74.56(C-5″), 74.51(C-2″), 74.04(C-4), 73.12(C-3′), 72.21(C-6), 70.70(C- 3″), 70.52(C-6′), 66.96(C-6″), 62.81(C-2′), 59.97(C-3), 58.70(s), 48.67(C- 1), 47.23(C-4′), 31.82(C-2), 20.80(Ac), 15.51(CH3-C-6′), 14.05(CH3-C-5″).

[0993] MALDI TOFMS: C 21 H 40 N4O 11 ([M+Na] + )m / e 602.56; measured value is m / e 625.23.

[0994] 6′-(R)-methyl-5-O-(5″-(S)-azido-5″-deoxy-β-D-ribofuranose)-2′,3-azido-1-N-benzamide paromomamide (Compound 25; Figure 5 ) Preparation:

[0995]

[0996] Compound 25 was prepared as described for the preparation of compound 19 using compound 18 (0.422 g, 0.405 mmol) and a solution of MeNH2 (33% solution in EtOH, 10 ml) to give 0.260 g (96%).

[0997] 1H NMR (500MHz, MeOD): Ring I: δ=6.11 (d, 1H, J=3.5Hz, H-1), 4.07-4.03 (m, 1H, H-6), 4.01-3 .93 (m, 2H, H-3, H-5), 3.33 (dd, 1H, J=10.0, 8.8Hz, H-4), 3.14-3.07 (m, 1H, H-2), 1.24 (d, 3H, J=6.1Hz, CH3-C-6). Ring II: δ=4.04 (ddd, 1H, J=11.1, 10.0, 3.8Hz, H-1), 3.80-3. 72 (m, 1H, H-4), 3.72-3.65 (m, 1H, H-5), 3.65-3.55 (m, 1H, H-3), 3.51 (dd, 1H, J=10.6, 8.2Hz, H-6), 2.23 (dt, 1H, J=11.8, 3.6Hz, H-2eq), 1.57-1.47 (m, 1H, H-2ax). Ring III: δ= 5.33 (s, 1H, H-1), 4.17 (d, 1H, J=5.2Hz, H-2), 4.14-4.07 (m, 1H, H-3), 3.76-3.72 (m, 1 H, H-4), 3.66 (dd, 1H, J = 6.4, 5.6 Hz, H-5), 1.35 (d, 3H, J = 7.0 Hz, CH3-C-5′). Additional peaks in the spectrum were identified as follows: δ = 7.84 (d, 2H, J = 8.6 Hz, Ar), 7.54 (t, 1H, J = 7.4 Hz, Ar), 7.46 (t, 2H, J = 7.6 Hz, Ar).

[0998] 13 C NMR (126MHz, MeOD): δ=169.04 (carbonyl), 134.16 (Ar), 131.53 (Ar), 128.05 (Ar), 126.97 (Ar), 10 9.54(C-1″), 95.76(C-1′), 85.02(C-5), 84.82(C-4″), 75.23(C-2″), 75.15(C-4), 74.46(C -3′), 73.72(C-4′), 72.86(C-6), 71.32(C-3″), 71.16(C-5′), 67.61(C-6′), 63.44(C-2′), 60.69(C-3), 59.34(C-5″), 49.85(C-1), 32.36(C-2), 16.16(CH3-C-6′), 14.63(CH3-C-5′).

[0999] MALDI TOFMS: C 26 H 36 N 10 O11 ([M+Na] + )m / e 664.62; measured value is m / e 687.25.

[1000] 6′-(R)-methyl-5-O-(5″-amino-5″-deoxy-β-D-ribofuranose)-2′,3-amino-1-N-phenylsulfonamide paromycinamine (NB74-N1PhS; Figure 2 and 5 ) Preparation:

[1001]

[1002] Compound 19 (0.148 g, 0.243 mmol) was dissolved in a mixture of THF (5 ml) and aqueous NaOH solution (0.1 M, 1.5 ml). The mixture was stirred at room temperature for 10 minutes, after which PMe3 (1 M solution in THF, 4 ml) was added. The propagation of the reaction was monitored by TLC [CH2Cl2 / MeOH / H2O / MeNH2 (30% solution in EtOH), 10:15:6:15], indicating completion after 5 hours. The reaction mixture was then purified by flash chromatography on a silica gel column. The column was washed with EtOAc, THF, and MeOH. The product was eluted with a mixture of 20% MeNH2 solution in 80% MeOH (30% solution in EtOH). The fractions containing the product were combined and evaporated under vacuum; redissolved in a small amount of water and passed through Amberlite CG50 (NH4 + The column was first washed with H2O, MeOH / H2O, MeOH, and H2O. The product was then eluted with a mixture of MeOH / H2O / NH4OH (80:10:10) to give compound NB74-N1PhS (67 mg, 51%). For storage and biological testing, NB74-N1PhS was converted to its sulfate form: the free base was dissolved in water, the pH was adjusted to 7 with H2SO4 (0.1N) and lyophilized.

[1003] 1H NMR (500MHz, MeOD): Ring I: δ=5.16 (d, 1H, J=3.3Hz, H-1), 4.04 (dd, 1H, J=5.1, 2.2Hz, H-1 6), 3.74 (dd, 1H, J=9.9, 3.1Hz, H-5), 3.52-3.45 (m, 1H, H-3), 3.19-3.13 (m, 1H, H-4), 2.59 (q, 1H, J=6.2Hz, H-2), 1.15 (d, 3H, J=5.9Hz, 1H, CH3-C-6). Ring II: δ3.49 (t, 1H, J=9 .1Hz, H-5), 3.35-3.27(m, 2H, H-4, H-6), 3.17-3.08(m, 1H, H-1), 2.74-2.64(m, 1H, H- 3), 1.84 (dd, 1H, J=8.7, 4.0Hz, H-2), 1.23 (m, 1H, H-2) Ring III: δ=5.17 (d, 1H, J=2.2Hz, H -1), 4.06 (dd, 1H, J=6.6, 3.1Hz, H-2), 3.91-3.88 (m, 1H, H-3), 3.88-3.83 (m, 1H, H-4) , 2.96 (dd, 1H, J = 13.2, 3.5 Hz, 1H, H-5), 2.77 (dd, 1H, J = 13.2, 7.5 Hz, H-5). Additional peaks in the spectrum were identified as follows: δ = 7.89-7.86 (m, 2H, Ph), 7.60-7.55 (m, 1H, Ph), 7.52 (dd, 2H, J = 10.2, 4.7 Hz, Ph).

[1004] 13 C NMR (126MHz, MeOD): δ = 143.12 (Ph 4°), 133.54(Ph), 130.15(Ph), 127.96(Ph), 111.18(C-1″), 101.60(C-1′), 85.81(C-5), 85.69(C-4), 83.47(C-4″), 76.73(C-6), 76.45(C-5′), 76.34(s ), 74.92(C-3′), 73.53(C-4′), 72.69(C-3″), 67.75(C-6′), 57.54(C-2′), 55 .53(C-1), 51.95(C-3), 44.78(C-5″), 36.75(C-2), 16.55(CH3-C-6′).MALDI TOFMSC 22 H 33 N7O 13 S([M+H] + )m / e 608.66; measured value is m / e 609.06).

[1005] 6′-(R)-methyl-5-O-(5″-amino-5″-deoxy-β-D-ribofuranose)-2′,3-amino-1-N-acetamidobaromomycin (NB74-N1Ac; Figure 2 and 5 ) Preparation:

[1006]

[1007] Compound NB74-N1Ac was prepared as described for the preparation of compound NB74-N1PhS using compound 20 (278 mg, 0.544 mmol), THF (5 ml), NaOH (0.1 M, 1.5 ml), and PMe3 (1 M solution in THF, 4 ml) to afford 74 mg (29%).

[1008] 1 H NMR (500MHz, MeOD): Ring I: δ=5.15 (d, 1H, J=3.0Hz, H-1), 4.09-4.01 (m, 1H, H-6), 3.77 (dd, 1H, J=9.9, 3.1Hz, H-5), 3.49 (td, 1H, J=10.0, 1.5Hz, H-3), 3 .15 (dd, 1H, J=9.5, 9.1Hz, H-4), 2.62-2.55 (m, 1H, H-2), 1.14 (d, 3H, J=5. 5Hz, CH3-C-6). Ring II: δ = 3.75-3.67 (m, 1H, H-1), 3.53-3.45 (m, 1H, H-5), 3.3 7-3.28 (m, 2H, H-4, H-6), 2.81-2.73 (m, 1H, H-3), 1.91-1.90 (m, 1H, H-2), 1.25-1.13 (m, 1H, H-2). Ring III: δ = 5.19 (d, 1H, J = 2.8 Hz, H-1), 4.04 (dd, 1H, J = 5.1, 2.3 Hz, H-2), 3.92-3.87 (m, 1H, H-3), 3.87-3.82 (m, 1H, H-4), 2.98-2.92 (m, 1H, H-5), 2.80-2.73 (m, 1H, H-5). The additional peaks in the spectrum are identified as follows: δ = 1.90 (s, 3H, Ac).

[1009] 13C NMR (126 MHz, MeOD): δ = 173.41 (amide), 111.13 (C-1″), 101.75 (C-1′), 86.18 (C-4), 86.05 (C-5), 83.65 (C-4″), 83.55 (C-3′), 76.82 (C-6), 76.49 (C-2″), 76. 29(C-5′), 73.55(C-4′), 72.63(C-3″), 67.68(C-6′), 57.62(C-2′), 52.26(C -3), 51.18(C-1), 44.73(C-5″), 30.78(C-2), 18.38(Ac), 16.47(CH3-C-6′).

[1010] MALDI TOF MS: C 20 H 32 N 10 O 11 ([M+H] + )m / e 510.54; measured value is m / e 511.14.

[1011] 6′-(R)-methyl-5-O-(5″-amino-5″-deoxy-β-D-ribofuranose)-2′,3-amino-1-N-methanesulfonamide paromycinamine (NB74-N1MeS; Figure 2 and 5 ) Preparation:

[1012]

[1013] Compound NB74-N1MeS was prepared as described for the preparation of compound NB74-N1PhS using compound 21 (0.426 g, 0.68 mmol), THF (10 ml), aqueous NaOH (0.1 M, 2 ml), and PMe3 (1 M solution in THF, 2 ml) to give 150 mg (40%).

[1014] 1H NMR (500MHz, MeOD): Ring I: δ=5.23 (d, 1H, J=2.8Hz, H-1), 4.12 (dd, 1H, J=6.2, 2.6Hz, H-6), 3.82 (dd, 1H, J=10.0, 2.9Hz, H-5), 3.58-3.52 (m, 1H, H-3), 3.25-3.16 (m, 1H, H-4, 2.69-2.64 (m, 1H, H-4), 1.20 (d, 3H, J=1.0Hz, CH3- C-6). Ring II: δ=3.58-3.52 (m, 1H, H-5), 3.34 (dt, 2H, J=18.2, 8.3Hz, H-4, H-6 ), 3.28-3.21 (m, 1H, H-1), 2.90-2.78 (m, 1H, H-3), 2.15-2.09 (m, 1H, H-2eq), 1.38-1.29 (m, 1H, H-2, ax). Ring III: δ = 5.29 (d, 1H, J = 2.4 Hz, H-1), 4.13-4.10 (m, 1H, H-2), 3.98-3.94 (m, 2H, H-3, H-4), 3.09 (dd, 1H, J = 12.6, 2.6 Hz, H-5), 2.92-2.84 (m, 1H, H-5). The additional peaks in the spectrum are identified as follows: δ = 3.02 (s, 3H, NHSO2-CH3).

[1015] 13 C NMR (126MHz, MeOD): δ=109.99(C-1″), 100.09(C-1′), 84.92(C-5), 84.21(C -4), 81.48(C-3″), 75.33(C-3), 75.19(C-6′), 75.06(C-5′), 73.17(C-3′), 72.08(C-4′), 71.20(C-4″), 66.21(C-2″), 55.94(C-2′), 54.03(C-5″), 50. 58(C-1), 48.16(C-6), 42.93(NHSO2-CH3), 36.49(C-2), 15.03(CH3-C-6′).

[1016] MALDI TOFMS: C 19 H 38 N4O 12 S([M+H] + )m / e 546.59; measured value is m / e 547.2.

[1017] 6′-(R)-methyl-5-O-(5″-amino-5″-deoxy-β-D-ribofuranose)-2′,3-amino-1-N-benzamide paromycinamide (NB74-N1Bz; Figure 2 and 5 ) Preparation:

[1018]

[1019] Compound NB74-N1Bz was prepared as described for the preparation of compound NB74-N1PhS using compound 22 (0.25 g, 0.38 mmol), THF (5 ml), NaOH (0.1 M, 1.5 ml), and PMe3 (1 M solution in THF, 2 ml) to afford 114 mg (52%).

[1020] 1 H NMR (500MHz, MeOD) Ring I: δ=5.26 (d, 1H, J=2.8Hz, H-1), 4.16 (dd, 1H, J=7.0, 2.0Hz, H-6), 3.87 (dd, 1H, J=9.9, 3.0Hz, H-5), 3.60 (t, 1H, J=9.5Hz, H-3), 3.30-3.23 (m, 1H, H-4), 2.67 (dd, 1H , J=10.3, 4.7Hz, H-2), 1.23 (d, 1H, J=5.1Hz, CH3-C-6). Ring II: δ=4.10-4.02 (m, 1H, 3-H), 3.63 (ddd, 2H, J=5.9, 2.0, 1.0Hz, H-4, H-5), 3.47-3.41 (m, 1H, H-6), 2.97-2.88 (m, 1H, H-1), 2.09 (ddd, 1H, J=3.8, 2.5, 0.7Hz, H-2eq), 1.43 (dt, 1H, J=26.9, 6.7Hz, H-2ax). Ring III: δ=5.32 (d, 1H, J=2.0Hz, H-1), 4.14 (dd, 1H, J=5.3, 2.1Hz, H-2), 4.00-3.96 (m, 1H, H-3), 3.93 (dd, 1H, J= 5.4, ​​3.8 Hz, H-4), 3.01 (dd, 1H, J = 13.4, 4.0 Hz, H-5), 2.84 (dd, 1H, J = 12.8, 7.7 Hz, H-5). Additional peaks in the spectrum were identified as follows: δ = 7.84 (d, 2H, J = 7.4 Hz, Ph), 7.52 (t, 1H, J = 7.4 Hz, Ph), 7.44 (t, 2H, J = 7.5 Hz, Ph).

[1021] 13C NMR (126 MHz, MeOD): δ = 168.98 (amide), 134.34 (Ph), 131.24 (Ph), 128.07 (Ph), 127.02 (Ph), 109.66 (C-1″), 100.42 (C-1′), 84.93 (C-4), 84.66 (C-6), 82.41 (C-4″), 75.39 (C-5′) , 75.04(C-2″), 74.60(C-4), 73.65(C-3′), 72.19(C-4′), 71.26(C-3″), 66.35(C-6′), 56.26(C-2′), 50.93(C-1), 50.36(C-3), 43.50(C-5″), 34.53(C-2), 15.18(CH3-C-6′).

[1022] MALDI TOFMS: C 25 H 40 N4O 11 ([M+H] + )m / e 572.27; measured value is m / e 573.22.

[1023] 6′-(R)-methyl-5-O-[5″-(S)-amino-5″-deoxy-β-D-ribofuranose]-2′,3-amino-1-N-benzamide paromycinamine (NB124-N1MeS; Figure 2 and 5 ):

[1024]

[1025] Compound NB124-N1MeS was prepared as described for the preparation of compound NB74-N1PhS using compound 23 (0.230 g, 0.358 mmol), THF (5 ml), aqueous NaOH (0.1 M, 2 ml), and PMe3 (1 M solution in THF, 2 ml) to give 117 mg (58%).

[1026] 1H NMR (500MHz, MeOD): Ring I: δ=5.22 (d, 1H, J=3.2Hz, H-1), 4.16-4.04 (m, 1H, H-4), 3.81 (dd, 1H, J=9.9, 3.1Hz, 1HH-5), 3.53-3.48 (m, 1H, H-3), 3.21 (dd, 1H, J=10.2, 8.7Hz, H-6), 2.62 (dd, 1H, J=10.4, 4.4Hz, H-2), 1.20 (d, 3H, J=5.8Hz, CH3-C-6). Ring II: δ=3. 54(t, 1H, J=9.1Hz, H-5), 3.35 (dt, 2H, J=13.4, 9.3Hz, H-4, H-6), 3.28-3.20 (m, 1H, H -1), 2.86-2.77 (m, 1H, H-3), 2.11 (dd, 1H, J=8.5, 4.1Hz, H-2eq), 1.33 (dd, 1H, J=26 .2, 12.4Hz, H-2ax). Ring III: δ = 5.26 (d, 1H, J = 2.6Hz, H-1), 4.08 (dd, 1H, J = 5.4, 2.4Hz, H-2), 4.04-3.98 (m, 1H, H-3), 3.59 (dd, 1H, J = 7.3, 6.3 Hz, H-4), 2.99 (t, 1H, J = 7.1 Hz, H-5), 1.20 (d, 3H, J = 7.1 Hz, CH3-C-5′). The additional peaks in the spectrum are identified as follows: δ = 3.03 (s, 3H, NHSO2-CH3).

[1027] 13 C NMR (126MHz, MeOD): δ=108.75(C-1″), 100.26(C-1′), 86.48(C-4″), 84.55(C-4) , 83.75(C-5), 75.31(C-6), 74.99(C-2″), 74.97(C-5′), 73.88(C-3′), 72.15(C- 6′), 71.07(C-3″), 66.36(C-4′), 56.22(C-2′), 54.15(C-1), 50.65(C-3), 49.56 (C-5″), 40.39(NHSO2-CH3), 36.69(C-2), 17.29(CH3-C-5′), 15.14(CH3-C-6′).

[1028] MALDI TOFMS: C 20 H 40 N4O 12 S([M+H2O] + ), m / e 560.24; measured value, m / e 561.2.

[1029] 6′-(R)-methyl-5-O-(5-(R)-amino-5-deoxy-β-D-ribofuranose)-2′,3-amino-1-N-acetamidobaromomycin (NB124-N1Ac; Figure 2 and 5 ) Preparation:

[1030]

[1031] Compound NB124-N1Ac was prepared as described for the preparation of compound NB74-N1PhS using compound 24 (0.820 g, 1.36 mmol), THF (10 ml), aqueous NaOH (0.1 M, 2 ml), and PMe3 (1 M solution in THF, 2 ml) to afford 370 mg (52%).

[1032] 1 H NMR (500MHz, MeOD): Ring I: δ=5.21 (d, 1H, J=4.8Hz, H-1), 4.07 (qd, 1H, J=6.4, 2.8H z, H-6), 3.77 (d, 1H, J=12.1Hz, H-5), 3.47 (dd, 1H, J=10.4, 8.9Hz, H-3), 3.15 (t, 1H, J=9.4Hz, H-4), 2.60 (dd, 1H, J=10.0, 3.2Hz, H-2), 1.14 (d, 3H, J=6.7Hz, CH3- C-6). Ring II: δ=3.71 (td, 1-H, J=12.3, 4.1Hz, H-5), 3.58-3.50 (m, 1H, H-6), 3.36 (dt , 2H, J=19.3, 9.4Hz, H-1, H-4), 2.82 (ddd, 1H, J=13.3, 9.7, 4.0Hz, H-3), 1.93 (dt , 1H, J=7.4, 4.0Hz, H-2eq), 1.23 (q, 1H, J=12.5Hz, H-2ax). Ring III: δ=5.22 (s, 1H, H- 1), 4.06 (dd, 1H, J=4.3, 2.3Hz, H-2), 4.00-3.95 (m, 1H, H-3), 3.64 (dd, 1H, J=8.9 , 5.8Hz, H-4), 3.11 (dd, 1H, J=7.5, 6.4Hz, H-5), 1.24 (d, 3H, J=8.0Hz, CH3-C-5′).

[1033] 13C NMR (126 MHz, MeOD): δ = 172.08 (amide), 109.46 (C-1″), 99.80 (C-1′), 85.02 (C-4), 84.22 (C-4″), 83.10 (C-3′), 75.41 (C-6′), 75.36 (C-5′), 74.93 (C-1), 73.27 (C-6) , 72.01(C-3″), 71.75(C-5″), 66.09(C-2″), 55.70(C-2′), 50.65(C-3), 50.29(C- 4′), 49.77(C-5), 34.16(C-2), 21.37(Ac), 15.49(CH3-C-5″), 14.93(CH3-C-6′).

[1034] MALDI TOFMS: C 21 H 40 N4O 11 ([M+H] + )m / e 524.56; measured value is m / e 525.26.

[1035] 6′-(R)-methyl-5-O-[5″-(S)-amino-5″-deoxy-β-D-ribofuranose]-2′,3-amino-1-N-benzamide paromycinamine (NB124-N1Bz; Figure 2 and 5 ) Preparation:

[1036]

[1037] Compound NB124-N1Bz was prepared as described for the preparation of compound NB74-N1PhS using compound 25 (0.260 g, 0.390 mmol), THF (10 ml), aqueous NaOH (0.1 M, 2 ml), and PMe3 (1 M solution in THF, 2 ml) to afford 160 mg (70%).

[1038] 1H NMR (500MHz, MeOD): Ring I: δ=5.22 (d, 1H, J=3.1Hz, H-1), 4.09 (dd, 1H, J=19.0, 8.6Hz, H-3), 3 .82 (dd, 1H, J=10.0, 3.0Hz, H-5), 3.53-3.47 (m, 1H, H-4), 3.19 (dd, 1H, J=10.1, 8.8Hz, H-6 ), 2.60 (dd, 1H, J=11.1, 5.1Hz, H-2), 1.18 (d, 3H, J=5.3Hz, CH3-C-6). Ring II: δ=4.05-3.95 (m , 1H, H-1), 3.57 (dd, 2H, J=7.6, 4.3Hz, H-6, H-5), 3.39 (dd, 1H, J=12.6, 6.1Hz, H-4), 2.92-2 .82 (m, 1H, H-3), 2.03 (dd, 1H, J=8.4, 4.0Hz, H-2eq), 1.37 (dd, 1H, J=25.9, 12.5Hz, H-2ax) .Ring III: δ=5.25 (d, 1H, J=2.3Hz, H-1), 4.07 (dd, 1H, J=5.4, 2.2Hz, H-2), 4.00-3.95 (m, 1H, H- 3), 3.58-3.53 (m, 1H, H-4), 2.98-2.91 (m, 1H, H-5), 1.16 (d, 3H, J = 7.2 Hz, CH3-C-5′). The additional peaks in the spectrum were identified as follows: δ = 7.81 (d, 2H, J = 7.3 Hz, Ar), 7.50 (t, 1H, J = 7.3 Hz, Ar), 7.43 (t, 2H, J = 7.5 Hz, Ar).

[1039] 13 C NMR (126MHz, MeOD): δ=168.99 (carbonyl), 134.36 (Ar), 131.22 (Ar), 128.05 (Ar), 126.98 (Ar), 10 8.81(C-1″), 100.33(C-1′), 86.56(C-4″), 84.92(C-4), 83.87(C-6), 75.36(C-5′), 75.05(C -2″), 74.46(C-5), 73.96(C-4′), 72.14(C-6′), 71.14(C-3″), 66.30(C-3′), 56.30(C-2′), 50.91(C-3), 50.36(C-1), 49.61(C-5″), 34.59(C-2), 17.34(CH3-C-5′), 15.06(CH3-C-6′).

[1040] MALDI TOFMS: C26 H 42 N4O 11 ([M+H] + )m / e 586.63; measured value is m / e 587.29.

[1041] An alternative synthetic route to prepare N1-sulfonyl-modified pseudodisaccharides, which can further serve as a catalyst for the synthesis of Figure 6 The intermediate A shown (see Figure 3 ) or compound 2 (see Figure 4 ) The acceptor compound of group 1 and group 2 compounds (when the hydroxyl protecting group is introduced at the corresponding position described herein) is represented as acceptor B* herein. The overall yield is 36%. The structures of intermediate compounds 82 and 83, and the obtained compound 84 were verified using NMR and MS measurements described herein.

[1042] Example 2

[1043] Activity and toxicity of the compound of Example 1

[1044] All aminoglycosides tested were used in the form of their sulfate salts. The molecular weights (g / mol) of the sulfate salts are as follows:

[1045] NB74-N1PhS-726.40, NB74-N1Ac-581.62, NB74-N1MeS-608.74, NB74-N1Bz-634.23, NB124-N1MeS-640.76, NB124-N1Ac-559.28, NB124-N1Bz-667.75.

[1046] Readout activity; in vitro studies:

[1047] The readout activities of Group 1 and Group 2 compounds in Example 1 above were tested by using the dual luciferase reporter assay system as described below.

[1048] DNA fragments derived from PCDH15, CFTR, and IDUA cDNAs, including the tested nonsense mutations or the corresponding WT codons, and 4 to 6 upstream and downstream flanking codons, were created by annealing the following complementary oligonucleotide pairs:

[1049] Usher syndrome (PCDH15):

[1050] p.R3Xmut / wt:

[1051] 5′-GATCCCAGAAGATGTTTT / CGACAGTTTTATCTCTGGACAGAGCT-3′; and

[1052] 5′-CTGTCAGAGATAAAACTGTCA / GAAACATCTTCTG-3′.

[1053] p.R245Xmut / wt:

[1054] 5′-GATCCAAAATCTGAATGAGAGGT / CGAACCACCACCACCACCCTCGA-GCT-3′; and

[1055] 5′-CGAGGGTGGTGGTGGTTGTTCG / ACCTCTCATTCAGATTTTG-3′.

[1056] Cystic Fibrosis (CFTR):

[1057] p.G542Xmut / wt:

[1058] 5′-TCGACCAATATAGTTCTTT / GGAGAAGGTGGAATCGAGCT-3′; and

[1059] 5'-CGATTCCACCTTCTCA / GAAGAACTATATTGG-3'.

[1060] The fragments were inserted in frame into the polylinker of the p2Luc plasmid between the restriction sites of BamHI and SacI (p.R3X and p.R245X) or SalI and SacI (all others). For in vitro readout experiments, the resulting plasmids supplemented with the aminoglycosides tested were transcribed and translated using the TNT Reticulocyte Lysate Quick Coupled Transcription / Translation System. After incubation at 30°C for 90 minutes, the luciferase reporter assay system (Dual Luciferase Reporter Assay System (Promega) was used. TM ) Luciferase activity was measured. Stop codon readout was calculated as described in Grentzmann et al., RNA 4, 479-486 (1998).

[1061] The data were presented in the R3X nonsense mutation (premature UGA C stop codon) implicated in Usher syndrome and the G542X (premature UGA G stop codon) implicated in cystic fibrosis. Fig. 7A -D.

[1062] like Fig. 7A-D, all tested aminoglycosides induced a readout, except for NB74-Bz and NB124-Bz (data not shown). Among the aminoglycosides tested, NB124-MeS induced the highest readout level, followed by NB74-MeS, at all tested concentrations. For the R3X nonsense mutation, NB74-MeS showed about 1.5-fold better readout activity than NB74; and NB124-MeS also exhibited superior readout activity to NB124. For the G542X mutation, compounds NB74-MeS and NB124-MeS showed better activity than NB74 and NB124, respectively, confirming the substantial impact of the methanesulfonyl moiety.

[1063] The data obtained indicate that methanesulfonyl substitution at the N1 position facilitates the interaction of NB compounds with the eukaryotic A site and induces their readout potency.

[1064] Protein translation inhibition test:

[1065] The ability of Group 1 and Group 2 compounds as described in Example 1 above to inhibit eukaryotic translation was further tested as follows.

[1066] The luciferase T7 control DNA plasmid (Promega) was used according to the manufacturer's protocol. The T7 fast coupled transcription / translation system quantifies eukaryotic in vitro translation inhibition. Translation reactions (25 μL) containing variable concentrations of the tested aminoglycosides were incubated at 30°C for 60 minutes, cooled on ice for 5 minutes, diluted with diluent, and transferred to 96-well plates. In prokaryotic and eukaryotic systems, luminescence was measured immediately after the addition of luciferase assay reagent (50 μL; Promega), and light emission was recorded with a FLx800 fluorescence microplate reader (Biotek). Half-maximal inhibitory concentration (IC50) values ​​were obtained by fitting the data of at least two independent experiments from concentration response curves using Grafit 5 software.

[1067] The obtained data are shown in 1.

[1068] Table 1

[1069]

[1070] As shown in Table 1, NB74-MeS (IC 50 =8.35μM) and its parent structure NB74 (IC 50 =14.31μM) 50 The values ​​indicate that NB74-MeS is 1.7 times more specific for eukaryotic ribosomes, while NB74-Bz (IC 50=754.77 μM) is 54-fold less specific for eukaryotic ribosomes than NB74. This overall data suggests that the observed effect of the methylsulfonyl group on the enhanced readout activity of NB74-MeS is associated with an increase in its specificity for eukaryotic ribosomes.

[1071] Activity readout for cell-free experiments:

[1072] Rabbit reticulocytes (TNT mixture) were used to transcribe and translate the dual luciferase plasmids carrying the cystic fibrosis transmembrane conductance regulator (CFTR) mutation, S466X, or wild-type sequence between Renilla and firefly luciferase. The WT plasmid expresses both firefly and Renilla luciferase, while the mutant plasmid only expresses Renilla luciferase due to the stop codon in the insertion sequence. By adding the compound to the in vitro transcription / translation reaction mixture, the test compound and the control were read out.

[1073] Cystic Fibrosis (CFTR):

[1074] p.S466Xmut / wt:

[1075] 5′-GGCAAGACTTGACTTCTAATGGTG-3′; and

[1076] 5'-GGCAAGACTTCACTTCTAATGGTG-3'.

[1077] The readout activities induced by NB74-MeS and NB124-MeS were calculated from the dual-luciferase assay data using the results from both WT and mutant plasmids and the following equation:

[1078]

[1079] The following parameters were also calculated from the dual luciferase assay results for WT and mutant plasmids:

[1080] (i) Translation inhibition (TI; μM): The degree of translation inhibition of ribosomes by NB74-MeS and NB124-MeS was calculated by nonlinear regression curve fitting on the plot of relative light units (RLU) versus log NB74-MeS and NB124-MeS molar concentrations using the reduction in Renilla bioluminescence values ​​of the WT plasmid (Find ECanything analysis, GraphPad PRISM, version 7).

[1081] (ii) NB74-MeS and NB124-MeS readout potency (EC50, μM): The readout activity of NB74-MES and NB124-MES is presented as firefly luciferase activity normalized to the Renilla luciferase activity in the mutant plasmid. Normalized firefly values ​​were plotted against the log molar NB74-MeS and NB124-MeS concentrations, and the curves were fitted using four-parameter nonlinear regression (GraphPad PRISM software, version 7). NB74-MeS and NB124-MeS cause translational inhibition at high concentrations, thus not allowing further dose increases. Therefore, to enable EC50 calculations, the range of values ​​was NB74-MeS and NB124-MeS concentrations less than TI80 (determined as above).

[1082] (iii) Readout efficiency is presented as the fold increase in firefly luciferase activity between NB74-MeS and NB124-MeS treated and untreated mutant plasmids. The fold increase was calculated as the concentration of NB74-MeS and NB124-MeS that showed ≤20% inhibition of steady-state Renilla activity on the wild-type control plasmid.

[1083] (iv) The % WT readout was calculated by dividing the percentage of FF / Renilla of the mutant plasmid by the percentage of FF / Renilla of the WT plasmid.

[1084] Incubation of NB74-MES and NB124-MES with nonsense mutations of cystic fibrosis S466X resulted in dose-response inhibition in cell-free experiments (data not shown). The calculated TI80 values ​​were 16 μM for NB74-MeS and 1 μM for NB124-MeS, respectively. The calculated TI50 values ​​were >16 μM for NB74-MeS and 2 μM for NB124-MeS, respectively.

[1085] Table 2 below shows the results of a dose-response cell-free assay for cystic fibrosis S466X nonsense mutation inhibition for two exemplary compounds NB74-MeS and NB124-MeS at concentrations ranging from 0-16 μM according to an embodiment of the present invention. The data shown in Table 2 show that incubation of increasing doses of NB74-MeS and NB124-MeS resulted in a substantial increase in readout (about 2-fold to about 10-fold) compared to the control (untreated cell extract), and 9-17% readout compared to the wild-type control plasmid.

[1086] Table 2

[1087]

[1088] *Relative to MUT control

[1089] **Relative to WT

[1090] These data further support the finding that the inclusion of a sulfonyl moiety at the N1 position has a significant impact on the biological activity of aminoglycosides.

[1091] Toxicity test on cochlear explants:

[1092] The ototoxic potential of Group 1 and Group 2 compounds was tested using cochlear explants as described, for example, in E. Shulman et al., J. Biol. Chem. 289(4), 2318-2330 (2014). Ototoxicity was measured as the concentration of AG required to shed 50% of the hair from cochlear outer hair cells, as described below.

[1093] The auditory hair cell toxicity of the test compounds was screened in explants of the organ of Corti of postnatal day 2 to 3 CBA / J mice. The excised tissue was placed on a collagen-coated incubation dish with a flat surface preparation of 1 ml of serum-free basal medium Eagle (Sigma Aldrich, St. Louis, MO) containing serum-free supplement (Invitrogen, Eugene, OR), 1% bovine serum albumin, 2 mM glutamine, 5 mg glucose / ml and 10 units of penicillin / ml. The explants were incubated for 5 hours (37°C, 5% CO2), and an additional 1 ml of medium was added to immerse the explants. After 48 hours of continuous preincubation, the medium was replaced with new medium containing the test compound at the specified concentration. After an additional 72 h of incubation, explants were washed three times with PBS, fixed in 4% (v / v) paraformaldehyde overnight at 4°C, and then permeabilized with 0.3% (v / v) Triton X-100 in PBS for 30 min. Specimens were washed three times with PBS for 10 min each at room temperature and then incubated with rhodamine phalloidin (1:100; Life Technologies, Eugene, OR) for 1 h at room temperature or overnight at 4°C.

[1094] After rinsing several times with PBS, the explants were mounted on microscope slides with Fluoro-Gel (Electron MicroscopySciences, Hatfield, PA) and imaged on a Leica SP5 confocal TCS microscope (Leica, Wetzlar, Germany). Hair cells were identified by phalloidin staining of ciliary hair bundles and peripheral F-actin rings. Their presence or absence was quantified using a 50x oil immersion objective on an epifluorescence Leitz Orthoplan microscope (Leica, Wetzlar, Germany) equipped with a calibrated ruler (0.19 mm) superimposed on the field of view. All hair cell rows were longitudinally oriented in each frame and counted from the apex to the base of the cochlea. Cell counts were entered into a computer program and compared with a standard database (KHRI Cytocochleogram, version 3.0.6, Kresge Hearing Research Institute, University of Michigan, Ann Arbor, MI, USA) and reported as the average percentage of hair cell loss over the entire length of the explant.

[1095] Figure 8 Table 3 below shows a comparative dose-response curve, indicating that for NB124-MeS, a 50% loss of hair cells (LC 50 Coch ), and for NB74-MeS, it is 61.77 μM. The value obtained for NB124-MeS is better than that for NB124 (LC 50 Coch =11.1 μM). The ototoxicity of NB124-MeS was almost two times lower than that of NB124.

[1096] It was found that both NB74-Ac and NB124-Ac exhibited incredibly low ototoxic potential (LC 50 Coch For example, in the case of acetate substitution of NB74, ototoxicity was reduced by three orders of magnitude (NB74-Ac LC 50 Coch =2.4mM; NB74LC 50 Coch =112 μM).

[1097] These data also show that for NB74-PhS, NB74-Ac, and NB124-Ac, the ototoxicity data correlates directly with the readout activity and inhibition of eukaryotic translation data. These compounds have low affinity for eukaryotic ribosomes and therefore have significantly reduced ototoxicity.

[1098] Table 3

[1099]

[1100] Overall, these data suggest that there should be a delicate balance between electronic and stearic interactions of substituents with the target MET channel that influences cochlear toxicity. However, the observed increased readout activity and reduced ototoxicity demonstrated for NB124-MeS are significant, suggesting it is a promising drug candidate.

[1101] The ototoxicity of NB124-MeS, NB124-Ac, and NB124 was further examined by measuring cochlear cell loss, and the resulting data are shown in Fig. 9A -D. As shown, staining for F-actin reveals an organized outline of three rows of outer hair cells (OHCs) and one row of inner hair cells (IHCs) starting from the root of the cochlea (see Fig. 9A In the presence of 15 μM NB124-MeS, the structure of the epithelium remained normal and was almost indistinguishable from the control, with an average damage of 10% (see Fig.9D In contrast, in the presence of 15 μM NB124, hair cell loss was almost complete, showing over 80-90% cell loss (see Fig. 9B ). For NB124-Ac at a concentration of 150 μM, the epithelium remained normal and indistinguishable from the control (see Fig. 9C ).

[1102] The data obtained in these studies indicate that the substitution of the methanesulfonyl group at the N1 position contributes significantly to the readout activity of the compound. In addition, NB124-MeS exhibits a significant reduction in the level of ototoxicity compared to the parent NB124.

[1103] Example 3

[1104] Some exemplary 6'-modified compounds according to the present embodiments

[1105] The inventors have performed some modeling studies based on the recently solved structure of G418 bound to the leishmania A site rRNA sequence (Shalev et al. PNAS 110, 13333-338, (2013)), and the structure of G418 bound to the yeast ribosome (M. Yusopov et al. Nature 513, 517-22 (2014)). These studies suggest that the conversion of the C6'-OH of ring I to the corresponding carboxylic acid or amide (C(=O)OH and / or C(=O)NH2) may bring the ring I of the aminoglycoside closer to G1408 in mammalian ribosomes through strong electrostatic and / or H-bonding interactions between the carboxylate / amide and the G1408 residue. This hypothesis is further supported by experimental data showing that compounds with 6'-NH2 (such as neomycin and gentamicin) are inactive against Leishmania and that their activity / binding affinity to mammalian ribosomes is significantly lower than that of compounds with 6'-OH (such as paromomycin and G418) (Shalev et al. PNAS 110, 13333-338, (2013)), which is explained by the electrostatic repulsion between 6'-NH3+ (ammonium) and G1408; and by experimental data showing that such pseudodisaccharide compounds characterized by C(=O)OH or C(=O)NH2 at the C6' position were previously synthesized (Simonson et al., ChemBioChem 3, 1223-28, 2002) and lack any antibacterial activity, with an IC50 value against prokaryotic ribosomes that is about two orders of magnitude weaker than the parent paromomycin (6'-OH).

[1106] Although such compounds have not been studied in the context of mammalian ribosomes, the present inventors have designed and synthesized pseudodisaccharide and pseudotrisaccharide compounds, e.g. Fig.10 As shown, their interaction with residue G1408 in the mammalian cytoplasmic A site was investigated.

[1107] Fig.11 The general synthetic pathways for the pseudodisaccharide aminoglycosides NB160 and NB161 are shown, as Fig.10 shown.

[1108] Down Fig.12 The general synthetic route of the newly designed pseudotrisaccharide aminoglycosides NB162-165 is shown, as Fig.10 shown.

[1109] As described in Example 2 above, Fig.10 Preliminary readout activity testing of the indicated compounds on the R3X mutation indicated that these compounds exhibited low readout activity (data not shown).

[1110] This was also confirmed by experiments with eukaryotic translation systems performed as described above in Example 2. The IC50 values ​​measured for the tested compounds on the mammalian translation system showed poor inhibition (data not shown).

[1111] In a series of modeling, docking and molecular simulation studies, we found that in compounds featuring a carboxylic acid at the C6' position, an internal salt bridge is formed between the carboxylate and the N5"-ammonium, which significantly transforms the binding interaction of the ligand with the RNA host.

[1112] Example 4

[1113] Some exemplary 4'-modified compounds according to the present embodiments

[1114] While continuously trying new structural designs of aminoglycosides in order to eliminate the ototoxic effect while retaining its effective readout activity, the inventors have considered the reduced ototoxicity of recently reported paromomycin derivatives, which are modified at the O4' or O4' and O6' positions (see Perez-Fernandez, D. Commun. 5, 3112 (2014); Akbergenov R. et al. Am. Soc. Microbiol. 5, 1-10 (2014)). It is noteworthy that similar 4'-O-alkylation of kanamycin did not result in increased ribosomal selectivity and reduced toxicity, as in the case of paromomycin (see Kato T. et al. ACS Infect. Dis. 1, 479-486 (2016)).

[1115] The present inventors have designed a series of compounds based on the previously described NB74 and NB124 pseudotrisaccharides, which are characterized by modifications at the O4' position (also referred to herein as Group 4 compounds) or modifications at the O4' and O6' positions (also referred to herein as Group 3 compounds), such as Fig.13 As shown, each is optionally combined with the N1-substituent described in Example 1 above (e.g., N1-methylsulfonyl substitution), such as Fig.14 (also referred to herein as Group 5 and Group 6 compounds).

[1116] like Fig.13 As shown, the newly designed structures include, for example, modifications of the parent NB74 and NB124 having one of the 4',6'-ethylene groups (compounds 61 and 63), or 4',6'-benzylidine (compounds 62 and 64) in group 3, or 4'-O-ethyl groups (compounds 65 and 67), or 4'-O-propyl groups (compounds 66 and 68) in group 4.

[1117] Fig.15 A general synthetic route to Group 3 compounds is shown.

[1118] Briefly, for the synthesis of Group 3 structures, commercially available G418 was converted to intermediate A (see Figure 3 ). After the formation of cyclic acetals at the 4',6'-hydrozyls, selective acetylation was performed to give acceptor C. After coupling with a trichloroacetamide donor as described previously (Nudelman et al., 2006, supra), two deprotection steps were performed to give Group 3 structures.

[1119] Fig.16 An exemplary method for preparing an exemplary acceptor C, wherein Rw is Ph (Compound 72) or p-methoxyphenyl (PMP; Compound 74), is shown and described below.

[1120] Briefly, the common intermediate 2 was obtained from commercially available G418 in two steps (acid hydrolysis and perazidation) as described previously [Nudelman et al., Bioorganic Med. Chem. 18, 3735-3746 (2010)]. The cyclic acetal was then formed at the 4', 6'-hydrozyls by selective acetylation with benzaldehyde dimethyl acetal or p-methoxybenzaldehyde dimethyl acetal to give selectively protected 71 or 73, respectively. These two acetals were then converted to the corresponding common acceptors 72 and 74, respectively, by reaction with acetic anhydride at low temperature.

[1121] Donors 75 and 76 (such as Fig.16 ) was prepared as described previously [Joseph et al., Chem. Commun. 51, 104-106 (2015)].

[1122] The coupling of each acceptor 72 and 74 to the donor 75 or donor 76 is then completed using the aforementioned steps.

[1123] A similar sequence of steps was used for the assembly of group 4 structures, such as Fig.17 shown.

[1124] Briefly, intermediate A was first converted to intermediate D by using a sequence of steps similar to those reported for paromomycin (Perez-Fernandez, D. et al. Nature Commun. 5, 3112 (2014); Akbergenov, R. et al. Am. Soc. Microbiol. 5, 1-10 (2014)). Intermediate D was then converted to the common acceptor E by various alkylations at the C4′ position using CAN or DDQ and selective removal of PMB protection, followed by acetylation to give the desired acceptor containing C5-OH without ring III attached. Coupling of a trichloroacetimidate donor and two deprotection steps then gave Group 4 compounds.

[1125] exist Fig.18A and 18B An alternative synthetic route is shown in . Briefly, cyclic acetals 71 and 73 (see Fig.16 ) were first converted into fully protected intermediate acetals 77 and 79, which were then subjected to selective ring-opening of the cyclobenzylidene acetal to afford compounds 78 and 80, respectively.

[1126] In a fork-displacement step, cyclic acetals 71 and 73 are removed to generate the corresponding 4',6'-diols, followed by selective protection of the exocyclic 6'-OH with a bulky protecting group that is stable under basic conditions required for introduction of the alkyl group at the 4' position.

[1127] Fig.14 Chemical structures are presented that combine structural modifications introduced into Group 1 and Group 2 as described herein (e.g., N1-SMe modifications) with structural modifications in Group 3 and Group 4 as described herein, thereby providing compounds collectively referred to herein as Group 5 and Group 6 structures.

[1128] Fig.19 General synthetic routes for preparing Group 5 and Group 6 compounds are given.

[1129] Briefly, commercially available G418 is first converted to intermediate F, which contains a methanesulfonyl substitution at the N1 position of ring II, as described above in Example 1. Next, intermediate F is further alkylated to produce the common acceptor G and / or H series, which is then coupled with a trichloroacetimidate donor of the third ring, as described previously. This two-step deprotection then provides compounds of Groups 5 and 6.

[1130] Fig. 20 An exemplary synthetic route for preparing an exemplary receptor G, compound 86, is shown.

[1131] In short, as mentioned above and Figure 6As shown, amine 2 was prepared and then converted to the corresponding N1-methanesulfonate 84. Benzylydene protection was performed to give compound 85, followed by selective protection of the alcohol to give compound 86, with good isolated yield as exemplary receptor G. Receptor H was similarly prepared from, for example, compound 84. Using compounds NB74 and NB124 as parent reference compounds, compounds of Groups 3, 4, 5 and 6 (stored as their sulfate salts, as described in Example 1 above) were subjected to comparative readout activity experiments, metastasis inhibition experiments and toxicity tests, as described in Example 2 above. Compounds that exhibited good readout activity and low cytotoxicity were subsequently subjected to cochlear toxicity tests, as described in Example 2 above.

[1132] Although the present invention has been described in conjunction with specific embodiments thereof, it is apparent that many alternatives, modifications and variations will be apparent to those skilled in the art. Therefore, it is intended to encompass all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

[1133] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety, to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application should not be construed as an admission that the reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

Claims

1. A compound represented by the general formula I: or a pharmaceutically acceptable salt thereof, in: The dotted line indicates the stereo configuration of the 6′ position of the R configuration or the S configuration; R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl; R2 is selected from hydrogen, substituted or unsubstituted alkyl and ORx, wherein Rx is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, or alternatively, R2 is said ORx and together with R3 forms dioxane; R3 is selected from hydrogen, substituted or unsubstituted alkyl and ORy, wherein Ry is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkaryl, and acyl, or alternatively, R3 is said ORy and together with R2 forms dioxane; R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl, Provided that at least one of R7-R9 is a sulfonyl group.

2. The compound according to claim 1, wherein R7 is the sulfonyl group, and the compound is represented by Formula Ia: in: R1-R6, R8 and R9 are as defined in Formula I; and R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

3. A compound represented by the general formula I*: or a pharmaceutically acceptable salt thereof, in: The dotted line indicates the stereo configuration of the 6′ position of the R configuration or the S configuration; R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl; R2 is ORx, wherein Rx is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl; R3 is ORy, wherein Ry is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted alkaryl; R4-R6 are each independently selected from hydrogen, substituted or unsubstituted alkyl, and ORz, wherein Rz is selected from hydrogen, a monosaccharide moiety, an oligosaccharide moiety, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted alkaryl, and an acyl group; and R7-R9 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl and sulfonyl, And wherein the ORx and the ORy are connected to each other so that R2 and R3 together form dioxane.

4. The compound according to claim 3, represented by formula I*a: in: R1, R4-R6 and R7-R9 are as defined in Formula I*; and Rw is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

5. The compound according to claim 3, wherein R7 is the sulfonyl group, and the compound is represented by formula I*b: in: Rw, R1, R4-R6, R8 and R9 are as defined in Formula I*; and R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

6. The compound according to any one of claims 1-5, wherein at least one of R4-R6 is ORz and Rz is the monosaccharide moiety or the oligosaccharide moiety.

7. The compound according to any one of claims 1-5, wherein R5 is ORz and Rz is the monosaccharide moiety.

8. The compound according to any one of claims 1 to 7, wherein the monosaccharide moiety is represented by Formula II: The curved lines indicate the locations of the connections; The dotted line indicates the stereo configuration of the 5″ position of the R configuration or the S configuration; R 10 and R 11 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, and acyl; R 12 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl; and R 14 and R 15 are each independently selected from hydrogen, acyl, amino-substituted α-hydroxyacyl, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted alkaryl, sulfonyl and a cell-permeable group, or alternatively, R 14 and R 15 Together they form a heterocyclic ring.

9. The compound according to any one of claims 1-2, wherein R5 is ORz and Rz is the monosaccharide moiety represented by formula II, and the compound is represented by formula III: in: R1-R4 and R6-R9 are each as defined in Formula I or Formula Ia; and R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in formula II in claim 21.

10. The compound according to claim 9, wherein R7 is the sulfonyl group, and the compound is represented by formula IIIa: in: R1-R4, R6, R8 and R9 are as defined in Formula I or Formula Ia; R 10 , R 11 , R 12 , R 14 and R 15 As defined in formula II of claim 21; and R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

11. The compound according to claim 10, wherein R 20 , R 11 , R 14 and R 15 Each is hydrogen and R 12 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted aryl.

12. A compound according to any one of claims 3 to 5, wherein R5 is ORz and Rz is the monosaccharide moiety represented by formula II, and the compound is represented by formula III*: in: R1-R4 and R6-R9 are each defined as in formula I* or I*a or I*b; and R 10 , R 11 , R 12 , R 14 and R 15 Each is as defined in formula II in claim 21.

13. The compound according to claim 12, wherein the dioxane is a substituted or unsubstituted 1,3-dioxane, and the compound is represented by formula III*a: in: R1, R4, R6, and R7-R9 are as defined in formula I* or I*a or I*b; R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and Rw is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

14. The compound according to claim 13, wherein Rw is selected from substituted or unsubstituted alkyl and substituted or unsubstituted aryl.

15. A compound according to any one of claims 13-14, wherein R7 is the sulfonyl group, and the compound is represented by formula III*b: in: Rw, R1, R4, R6, R8 and R9 are as defined in formula I*b; R 10 , R 11 , R 12 , R 14 and R 15 As defined in Formula II; and R' is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkaryl, and substituted or unsubstituted aryl.

16. The compound according to any one of claims 1 to 15, wherein R1 is substituted or unsubstituted alkyl.

17. A pharmaceutical composition comprising the compound according to any one of claims 1 to 16 and a pharmaceutically acceptable carrier.

18. A compound according to any one of claims 1 to 16 or a composition according to claim 17 for use in treating a genetic disorder.

19. The compound or composition of claim 18, wherein the genetic disorder is associated with a premature stop codon mutation and / or a protein truncation phenotype.

20. The compound of any one of claims 1-16 or the composition of claim 17, for use in increasing the expression level of a gene having a stop codon mutation.

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