Linezolid derivatives and applications thereof

Abstract

The present disclosure relates to linezolid derivatives, in particular of Formula (I), the definition of p, A, A1 and R1 are as defined in the specification; the process of preparing the same and uses thereof in treatment of brain cancers, in particular glioblastoma.

Description

LINEZOLID DERIVATIVES AND APPLICATIONS THEREOFFILED OF THE INVENTION

[0001] The present disclosure relates to the pharmaceutical field. Particularly, the present disclosure pertains to linezolid derivatives, a process of preparing the same and uses thereof.BACKGROUND OF THE INVENTION

[0002] In the decades after approval of temozolomide (TMZ), no other effective drugs have been developed. Undoubtedly, blood-brain barrier (BBB) penetration is a key issue to overcome in glioblastoma mutiforme (GBM) drug development. The blood-brain barrier, the gatekeeper between the brain and circulatory system, resisted penetration of small-molecule drugs through blood vessels into the brain by nearly 100%, which presented an enormous obstacle in GBM drug development.

[0003] To date, the existing clinical technology still lacks an effective strategy to counter brain tumors, especially against the most aggressive type, glioblastoma mutiforme. Following approval of TMZ, the first medical treatment for GBM, a significant enhancement in overall survival propelled TMZ to deployment as first line therapy. Nevertheless, the hypermethylation caused by TMZ often leads to a frustrated outcome. According to statistics, more than 50% of GBM patients generated TMZ resistance after treatment. This may be due to overexpression of 06-methylguanine-DNA alkyltransferase (MGMT) and / or alkylpurine- DNA-N-glycosylase (APNG), activation of base excision repair (BER), or DNA mismatch repair (MMR) silencing.

[0004] In recent decades, researchers have sought solutions to recurrent GBM. Several strategies have been approved, including bevacizumab, nitrogen mustards, and other tumor treatments. However, these therapies still suffer certain crucial defects, for which there is urgent need for solution, including a lack of significant improvement in overall survival, potential systemic toxicity, and the limitations imposed by lesion location.SUMMARY OF THE INVENTION

[0005] The present disclosure relates to a novel therapeutic system for treating a cancer or a recurrent or metastatic cancer; particularly, GBM or recurrent or metastatic GBM.

[0006] The present disclosure provides a compound of Formula (I):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: p is 0 or 1, with the proviso that when p is 0, A has the definition of Ai, and when p is 1, A is acetyl;Ai has the formulathe formula of *-L-Rb, wherein * denotes the linking position to N;-L- is selected from the group consisting of-C U-, -C(=O)- and -S(=O)2-;-Sp- is selected from the group consisting of Ci-ealkylene,wherein ** denotes the linking position to L and ***denotes the linking position to the carbonyl in the moiety -C(=O)NHRa;Ra is -OH or 2-aminophenyl;Rb is selected from the group consisting ofand, wherein ** denotes the linking position to L; andRi is H, methyl or ethyl.

[0007] The subject disclosure also provides a compound of formula (IA):(IA), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein the definition of Ai and Ri are as noted for Formula (I); preferably, Ai is, wherein * denotes the linking position to N, -L- iswherein ** denotes the linking position to L and ***denotes the linking position to the carbonyl in the moiety -C(=O)NHRa; or preferably, Ai is, wherein * denotes the linking position to N, -L- is -C(=O)-, -Sp- is Ci-ealkylene, preferably Cealkylene, more preferably 1,6-w-hexanylene;or preferably,, wherein * denotes the linking position to N, -L-is -S(=O)2-, -Sp- is , wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa; or preferably, Ai is *-L-Rb, wherein * denotes the linking position to N, and L is -C(=O)-.

[0008] The subject disclosure also provides a compound of formula (IB):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein the definition of Ai and Ri are as noted for Formula (I); preferably, Ai is, wherein * denotes the linking position to N, -L- is-CH2-, -Sp- is wherein ** denotes thelinking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa; or preferably,, wherein * denotes the linking position to N, -L- is -C(=0)-, -Sp- is Ci-ealkylene, preferably Cealkylene, more preferably 1,6-w-hexanylene; or preferably,, wherein * denotes the linking position to N, -L-is -S(=O)2-, -Sp- is , wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa.

[0009] In one embodiments of Formula (I), (IA) or (IB), Ai iswherein * denotes the linking position to N, -L- is -CH2-, -Sp- is wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=0)NHRa, Ra is -OH and Ri is H.

[0010] In one embodiments of Formulawherein * denotes the linking position to N, -L- is -CH2-, -Sp- is wherein ** denotes the linking position to L and *** denotes the linking position to the carbonylin the moiety -C(=O)NHRa, Ra is -OH or 2-aminophenyl, preferably 2-aminophenyl, and Ri is H.

[0011] In one embodiments of Formula (I), (IA) or (IB), Ai iswherein * denotes the linking position to N, -L- is -C(=O)-, -Sp- is 1,6-w-hexanylene, Rais -OH and Ri is H.

[0012] In one embodiments of Formula (I), (IA) or (IB), Ai iswherein * denotes the linking position to N, -L- is -S(=O)2-, -Sp- is, wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa, Rais -OH and Ri is H.

[0013] In one embodiments of Formula (I), (IA) or (IB), Ai is *-L-Rb, wherein * denotes the linking position, -L- is -C(=O) and Rb iswherein ** denotes the linking position to L, and Ri is H.

[0014] In one embodiments of Formula (I), (IA) or (IB), Ai is *-L-Rb, wherein * denotes the linking position, -L- is -C(=O) and Rb is, wherein ** denotes the linking position to L, and Ri is H.

[0015] In one embodiments of Formula (I), (IA) or (IB), Ai is *-L-Rb, wherein * denotes the linking position, -L- is -C(=O) and Rb is, wherein ** denotes the linking position to L, and Ri is H, methyl or ethyl.

[0016] The compounds of Formula (I) include, but are not limited to:Compound (4)Compound (12)Compound (22)or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof.

[0017] The subject disclosure also provides a process of preparing a compound of Formula (I), Formula (I A) or Formula (IB).

[0018] The subject disclosure also provides a pharmaceutical composition comprising a compound as described herein, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, and a pharmaceutically acceptable excipient.

[0019] The subject disclosure also provides a method of treating a cancer or a recurrent or metastatic cancer, comprising administering an effective amount of a compound of Formula (I), Formula (IA) or Formula (IB), or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of foregoing to a subject in need.

[0020] In one embodiment, the compounds of Formula (I), Formula (IA) or Formula (IB) exhibit inhibitory effects on HD AC, preferably Class 1 HD AC.

[0021] In any previous aspects or embodiments, the compounds of Formula (I), Formula (IA) or Formula (IB) exhibit inhibitory effects on heat shock protein 90 (Hsp90) inhibitors.

[0022] In any previous aspects or embodiments, the compounds of Formula (I), Formula (IA) or Formula (IB) act as antitubulin agents.

[0023] In any previous aspects or embodiments, the compounds of Formula (I), Formula (IA) or Formula (IB) act as anti-cyclooxygenase (COX) agents, preferably anti-COX-2 agents.

[0024] In one embodiment, the compounds of Formula (I), Formula (IA) or Formula (IB) can cross the blood-brain barrier (BBB).

[0025] In one embodiment, the cancer is a brain cancer or a recurrent or metastatic brain cancer.

[0026] In a further embodiment, the brain cancer is glioblastoma (GBM) or a recurrent or metastatic GBM.BRIEF DESCRIPTIONS OF THE DRAWINGS

[0027] Fig. 1 shows exemplified compounds of Formula (I) of the present disclosure.

[0028] Figs. 2A-2D show inhibition of GBM and TMZ-resistant GBM growth by the compounds: (A) Structural representation of MPT1A089-098 and its inhibitory effects on A172R TMZ-resistant GBM cell line assessed using the MTT assay. (B) Comparison of cytotoxic efficiency between MPT1A089 and Linezolid using the MTT assay. (C) Evaluation of IC50 values for different GBM, TMZ-resistant GBM, and (D) primary culture astrocytes using the CCK8 assay.

[0029] Figs. 3A-3C showed that Compound (4) (MPT1A089) induces acetylation of Histone H3 and H4: (A) Analysis of RNA expression levels of Class I HDACs in non-tumor, GBM, and recurrent GBM, along with Kaplan-Meier survival curves, based on TCGA and CGGA data. (B) Western blot analysis reveals a significant induction of acetylation in Histone H3 and H4 within one hour of treating U87MGR GBM cells with 10 pM of MPT1A089. (C) Analysis of acetylation patterns at different sites on Histone H3 and H4.

[0030] Figs. 4A-4E show that Compound (4) (MPT1A089) inhibits the growth of TMZ- resistant GBM by suppressing DNA physiological functions: (A) Differential gene expression in PT#3R GBM cells treated with 10 pM of MPT1A089 for 72 hours. (B) Analysis based on Gene Ontology (GO) biological processes and (C) Gene Set Enrichment Analysis (GSEA)reveals significant inhibition of DNA replication and mitotic nuclear division, key DNA physiological functions. (D) Cell cycle analysis demonstrating that MPT1 A089 treatment leads to G2 / M phase arrest and an increase in the sub-Gl cell population in PT#3R cells. (E) Heatmap displaying the suppressed expression of genes related to DNA replication.

[0031] Figs. 5A-5E show that Compound (MPT1 A089) induces DNA repair dysfunction by reducing the stability of DNA repair proteins: (A) Homologous recombination (HR) assay conducted after treating with 2 and 2.5 pM of MPT1 A089 or DMSO for 48 hours. (B) Analysis of HR efficiency post-treatment, calculated as the percentage of HR by comparing MPT1 A089 and DMSO treatments, quantified after three independent experiments. (C) Treatment of PT#3- R GBM cells with MPT1 A089 at different concentrations for 72 hours. Western blot analysis of the protein expression levels of phospho- ATM, CtIP, RPA32, and RAD51. (D) PT#3-R cells treated with Cycloheximide, assessing the protein stability of CtIP and Rad51 in the presence and absence of MPT1A089. (E) Quantification based on the results of (D) after three independent experiments.

[0032] Figs. 6A-6G show that Compound (4) (MPT1A089) induces ubiquitination and degradation ofRad51 protein: (A) Immunoprecipitation (IP) conducted using a Rad51 antibody after 24 hours of MPT1A089 treatment in PT#3R GBM cells, confirming the ubiquitination- mediated degradation of Rad51 protein. (B) Schematic representation illustrating truncated Rad51 fragments based on different functional regions of the protein. (C) and (D) Overexpression of truncated Rad51 fragments in PT#3-R GBM cells, followed by immunoprecipitation with GFP and detection of ubiquitination on the fragments. (E) Plasmid constructs of Rad51 fragment 3 with lysine mutations to alanine. (F) and (G) Overexpression of the mutated fragment 3 in PT#3-R GBM cells, followed by immunoprecipitation with GFP and detection of changes in ubiquitination.

[0033] Fig 6H shows schematic diagram of RAD51 amino acid sequence truncation and design (upper) and schematic diagram of RAD51 amino acid sequence mutagenesis design(lower).

[0034] Figs. 7A-7E show penetration of Compound (4) (MPT1A089) and its Efficacy Against TMZ-Resistant GBM Cells: (A) UPLC signals corresponding to TMZ or MPT1 A089 detected in plasma and brain tissue extract after intraperitoneal injection, confirmed by (B) mass spectrometry as MPT1A089 signals. (C) MPT1A089 concentration in the samples and the Brain / Plasma (B / P) ratio. (D) Randomized grouping of mice with orthotopic PT#3-R cell transplantation, followed by intraperitoneal injections of DMSO, 10 mg / kg TMZ alone, or TMZ plus 10 mg / kg or 20 mg / kg MPT1A089 every three days from day 7, with Kaplan-Meier survival results. (E) Evaluation of the growth status of orthotopically transplanted tumors after treatment with TMZ alone or TMZ combined with MPT1 A089, assessed using H&E staining.DETAILED DESCRIPTION OF THE INVENTION

[0035] In order to facilitate understanding of the disclosure herein, terms as used herein are hereby defined below.

[0036] In the context of the specification and the claims, the singular forms "a," "an" and "the" include plural referents, unless specifically indicated otherwise. Unless otherwise stated, any and all examples and exemplary language (e.g., "such as") provided herein are merely used for better illustration of the present invention, instead of limiting the scope of the present invention.

[0037] It is to be understood that any numerical range recited in this specification is intended to include all sub-ranges encompassed therein. For example, a range from "50 to 70°C" includes all sub-ranges and specific values between the stated minimum value of 50°C and the stated maximum value of 70°C, inclusive, e.g. from 58°C to 67°C, and from 53°C to 62°C, 60°C or 68°C. Since the numerical ranges disclosed are continuous, they contain each numericalvalue between the minimum and maximum value. Unless otherwise specified, the various numerical ranges indicated in this specification are approximate.

[0038] In the present invention, the term "about" refers to an acceptable deviation of a given value measured by a person of ordinary skill in the art, depending, in part, on how to measure or determine the value.

[0039] In the present invention, the term "alkyl" refers to a saturated, straight or branched alkyl, which comprises preferably 1-10 carbon atoms, and more preferably 1-4 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, secbutyl, isobutyl, tert-butyl, 2-ethylbutyl, n-pentyl, isopentyl, 1 -methylpentyl, 1,3 -dimethylbutyl, n-hexyl, 1 -methylhexyl, n-heptyl, isoheptyl, 1,1,3,3-tetramethylbutyl, 1 -methylheptyl, 3- methylheptyl, n-octyl, 2-ethylhexyl, 1,1,3-trimethylhexyl, 1,1,3,3-tetramethylpentyl, nonyl, decyl, 1,1,3,3,5,5-hexamethylhexyl, or the like.

[0040] In the present invention, the term "alkoxyl" or "alkoxy" as used herein means a group having a formula "-O-alkyl," wherein the definition of the "alkyl" in said formula has the meaning of "alkyl" as stated above.

[0041] In the present invention, the term "cycloalkyl" as used herein means a saturated or partially unsaturated cyclic carbon radical containing 3 to 10 ring carbon atoms and more preferably 3 to 8 ring carbon atoms, and optionally an alkyl substituent(s) on the ring. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclopropenyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-cyclohexen-l-yl, and the like.

[0042] In the present invention, the term "halogen" or "halo" denotes fluorine, chlorine, bromine or iodine.

[0043] In the present invention, the term "amino" as used herein means a functional group of the formula -NR'R", wherein R' and R" each independently represent hydrogen or a hydrocarbyl group as defined above.

[0044] In the present disclosure, novel and potent compounds for crossing BBB are provided, and these compounds and pharmaceutical derivatives thereof may have potential for use in treating brain cancers, in particular GBM. The established delivery system was inspired by the commercially available drug linezolid, a synthetic antibiotic, which has been used as late-line therapy of the infection from multi-resistance gram-positive bacteria. Clinically, linezolid has been widely applied in the treatment of central nervous system (CNS) infections, wherein it has demonstrated promising penetration against the barrier between blood and cerebrospinal fluid (CSF), especially in meningitis patients. On the other hand, through targeting on the 50S ribosome in bacteria, the acting mechanism of linezolid is independent from affecting any of the homeostatic enzymes in Homo sapiens. Hence, the further repurposing of linezolid is expected to retain lipophilic barrier penetration ability with more predictable pharmacokinetic properties and acceptable safety profile.

[0045] To investigate the penetration property of linezolid carrier, several bioactive moieties were selected to achieve the desired modification. Histone modification is the one factor which crucial in altering not only cell-cycle modulation, differentiation, and apoptosis but also in tumor growth, progression and drug -resistance, and thus it reveals a potent target for GBM drug development. The downregulation of HD AC activities was expected to abolish or reduce TMZ-resistance. The repurposing of linezolid-conjugated HDAC inhibitors was anticipated to achieve high brain concentration over existing HDAC inhibitors with zinc- binding motif.

[0046] Beyond HDAC, heat shock protein 90 (Hsp90), a notable chaperone while involved in protein folding, enzymes assembly, and ligand binding, was considered as another potent therapeutic target. It is believed that the inhibition of Hsp90 not only provide cell stress to induce apoptosis, but the tumor microenvironment modulation ability possessed a dramatic synergistic effect with other chemotherapy agents, according to the applicant previous research. Treating onalespib, a synthetic resorcinol class Hsp90 inhibitor, silenced EGFR and itsdownstream pathway, including, AKT, ERK1 / 2, and ribosomal protein S6 kinase, in human glioma cell lines, as EGFR mutant was also found in GBM and known to enhance tumorgenecity. HSP90 might be a promising target for developing novel GBM therapeutic strategies.

[0047] Precision medicine has become the trend, under which development of target therapy has flourished; however, conventional chemotherapy is still an effective and comprehensive strategy for cancer treatment. Since the TMZ alkylating manner induced resistance, different strategies should be applied in development. Excluding direct modification on DNA, antitubulin agents have received promising feedback in treating various cancers, especially the two notable taxane drugs docetaxel and paclitaxel; however, their low BBB penetration limited their application to brain tumors. Due to the macro structure of taxane, further structural modification is limited. A conspicuous antitublin agent, colchicine, is believed to exhibit efficient BBB penetration, and the pharmacophore, the 2, 3, 4-trimethoxy phenyl moiety, may be applied.

[0048] Neuroinflammation facilitated drug resistance and progression in GBM, and the major cause of neuroinflammation was found to be the enhancement of arachidonate metabolism to form prostaglandins. Among the arachidonate metabolism pathways, phospholipids were initially converted to arachidonic acid by PLA2G5, and further catalyzed through cyclooxygenase-2 (COX-2), also known as PTGS2, for the conversion of prostaglandin H2 (PGH2). The prostaglandin H2 was then derived to be converted into prostaglandin F2a, E2, or D2, which is important for the downstream signaling. As a controller for synthesizing key intermediate PGH2, PTGS2 seems to be a potent target, and thus 5-chlorosalicylic acid may be incorporated into the structure for cyclooxygenase inhibition.

[0049] The present disclosure thus provides a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof as described herein.

[0050] Embodiments of the compounds of Formula (I) include the compounds with any combinations of the groups in Formula (I) as listed in the table below:wherein L, Sp, Raand Rb can be any combinations of the groups as listed in the table below:compound of Formula (IA) or Formula (IB) or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, as described herein. Embodiments of the compounds of Formula (IA) or Formula (IB) include any combinations of the groups as listed in the tables above.

[0052] In one embodiment, the compounds of Formula (I) include, but are not limited to Compounds (4), (8), (11), (12), (17), (18), (19), (22), (24), (26), (31), (34), (36) and (38), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein the structure of these compounds are as depicted above.

[0053] Process of preparing the compounds

[0054] In one embodiment, the process of preparing the compound of Formula (I) may be generalized to Scheme 1 or 2, with optional minor modification(s) such as protection / deprotection, etc.

[0055] Scheme 1 :

[0056] Reaction (1) involves using corresponding diacids, carboxyl-acyl halides, formylcarboxylic acids or esters (with subsequent conversion to carboxylic acid moiety) having the moiety of-L-Sp; reaction (2) is optional (if L needs to be reduced from -C(=O) to -CH2-) and involves using a suitable reducing agent such as NaHH reaction (3) is conducted when Rais -OH and involves using tetrahydropyranyl (THP)-protected hydroxylamine followed by deprotection, and optional methylation / ethylation to introduce Ri; reaction (4) is conducted when Rais 2-aminophenyl and involves using o-phenylenediamine, and optional methylation / ethylation to introduce Ri. Reaction (5) is conducted when A is *-L-Rb and involves using corresponding substituted benzylic acid or substituted benzoyl chloride, having the substitution pattern corresponding to Rb. The definitions of -L-, -Sp-, Ra and Rb are as described above.

[0057] Scheme 2:

[0058] Reactions (1) to (5) are similar to those described for Scheme 1.

[0059] The initial reactant in Scheme 2 may be prepared from the initial reactant used inScheme 1 via the following Scheme 3.

[0060] Scheme 3 :

[0061] Reagents and conditions: (a) AC2O, PhMe, reflux, 16h (b) HNO3, H2SO4, 0 °C to r.t.,Ih (c) H2, Pd / C, DCM, r.t., 16h.

[0062] Pharmaceutical Compositions and Medical Uses

[0063] The present disclosure also provides pharmaceutical compositions and applications or uses of the compounds described herein for the treatment of a cancer or a recurrent or metastatic cancer. The cancer can be, but is not limited to, a brain cancer, in particular glioblastoma.

[0064] The compounds described herein can be prepared as pharmaceutical compositions by methods which are commonly used, using excipients commonly used in this field, that is, pharmaceutical excipients, pharmaceutical carrier, or the like.

[0065] As a solid composition for oral administration, a tablet, powder, granule and the like is used. In such solid composition, one or two or more kinds of active ingredients are mixed with at least one inert excipient. The composition may contain an inert additive, for example, a lubricant, a disintegrant, a stabilizer, a solubilizer, and the like, added by means of commonly used methods.

[0066] A liquid composition for oral administration includes an emulsion, a solution preparation, a suspension, a syrup or an elixir, and the like which is pharmaceutically acceptable, and includes a generally used inert diluent, for example, purified water or ethanol. The liquid composition may contain adjuvants such as a solubilizing agent, a wetting agent, and a suspension, a sweetener, a flavor, an aromatic, or a preservative in addition to the inert diluent.

[0067] The solid composition or the liquid composition may also be encapsulated to form capsules, including but not limited to soft capsules or hard capsules.

[0068] The pharmaceutical composition may also be used via injection to a subject. The injection for parenteral administration includes a sterile aqueous or non-aqueous solution preparation, a suspension or an emulsion. As the aqueous solvent, for example, distilled water for injection or physiological saline is included. As the non-aqueous solvent, for example, alcohols such as ethanol are included. Such a composition may further include a tonicity agent, a preservative, a wetting agent, an emulsifier, a dispersant, a stabilizer, or a solubilizer. These are sterilized by, for example, filtration through a bacteria-retaining filter, mixing of a germicide,or irradiation. In addition, these can also be used in a manner in which a sterile solid composition is prepared, and is dissolved or suspended in sterile water or a sterile solvent for injection before being used.

[0069] A transmucosal agent such as a transnasal agent and the like is used in a solid, liquid, or semi-solid form, and can be prepared according to methods known in the related art. For example, a known excipient, a pH adjuster, a preservative, a surfactant, a lubricant, a stabilizer, a thickener, and the like may be suitably added. In administration, it is possible to use an appropriate device for inhalation or insufflation.

[0070] The following examples are provided to make the present disclosure more comprehensible to those of ordinary skill in the art to which the present disclosure pertains, but are not intended to limit the scope of the invention.EXAMPLES

[0071] Experiments and Materials

[0072] Chemicals and analytical methods

[0073] All chemicals were purchased from commercial suppliers and used without purification, except for compound 20, which was synthesized in the laboratory and used in the preparation of Compound (22), (24) or (26).

[0074] Melting points were determined on a Buchi 545 melting point apparatus. Nuclear magnetic resonance (XH NMR and13C NMR) spectra were acquired in CDCh, MeOD, or DMSO-de as indicated using Fourier NMR Spectrometer (Bruker Avance, Germany). 'H NMR spectral data are presented as illustrated: chemical shift (5) in ppm, multiplicity [s (singlet), d (doublet), t (triplet), m (multiple over the range specified)], number of protons, (nH), coupling constants (J) in hertz, and assignment of protons. High-resolution mass spectra (HRMS) were collected using an AB Sciex QStar XL electrospray ionization quadrupole time-of-flight mass spectrometry. The purity of the final compounds was determined by Shimadzu 2030C-NT HPLC system equipped with SPD-20A UV detector detecting at the wavelength of 254 nm.The separation was performed at Agilent ZORBAX Eclipse XDB-C18 column (4.6 mm x 150 mm, 5 pm) with a gradient elution A: acetonitrile B: water (lOmM ammonium acetate and 0.1% formic acid). The gradient program was as follows: 10% A (initial), 90% A (45 min), 10% A (50 min), 10% A (60 min). The flow rate was set at 0.5 mL / min, and the injection volume was 20 pL. All of the purity results were found to be > 95%.

[0075] Cell Culture and treatment

[0076] The TMZ-resistant human glioblastoma cell lines U87MG (HTB-14 ™), T98G (CRL-1690 ™), and A172 (CRL-1620 ™) were provided from the American Type Culture Collection (Manassas, VA, USA). The patient-derived cell line PT#3 was purified under the approval of the Institutional Review Board of the Taipei Medical University (No. 201006011). TMZ-resistant cell lines were established as described previously1. All cell lines were cultured in Dulbecco’s Modified Eagle Medium basal medium (Cat# 10-013 -CM, CORNING, MA, USA), supplemented with 10% fetal bovine serum (Cat# 12676029, Thermo Fisher Scientific, MA, USA), 100 units / mL penicillin, and lOOpg / mL streptomycin (Cat# 15140122, Thermo Fisher Scientific) at 37°C in a 95% air and 5% carbon dioxide environment, and maintained resistant characteristics with 50-100pg of temozolomide (Cat# HY-17364, MedChemExpress, NJ, USA ).

[0077] MTT assay

[0078] MTT assay was employed to evaluate the cytotoxicity of the synthesized compound against resistant GBM cells. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Cat# AM0818-0005) powder was purchased from BIONOVAS (Ontario, Canada). The MTT was prepared at 10 mg / mL stock with ultrapure water and then stored at -20°C. All cells were seeded at 2.5* 104per well in the 24-well plate for 24h, followed by treatment with different concentrations of compounds for 72h. Before the experiment, MTT stock was diluted 10-fold with culture medium and then incubated with cells for 30 minutes. After the incubation, the MTT medium was discarded, and 300 pL of dimethyl sulfoxide (DMSO, Cat# D2650,Sigma-Aldrich, MO, USA) was added to dissolve the cells. Finally, 100 pL of the mixture was transferred to measure the OD 595nm absorbance.

[0079] Cell proliferation assay

[0080] Cell viability and drug cytotoxicity were determined by a CCK-8 kit (Cell Counting Kit-8, CK04-13, MD, USA) (CCK8, C0005, Target Molecule Corp., MA, USA) reagent. Cells were seeded at 2* 103per well in the 96-well plate for 24h, followed by treatment with different concentrations of compounds for 72h. Before the experiment, CCK8 was diluted 10-fold with culture medium and then incubated with cells for 30 minutes. Finally, the OD 450 nm absorbance was measured and used to calculate cell viability.

[0081] HDAC Enzymes Inhibition Assays

[0082] Enzyme inhibition assays were performed by the Reaction Biology Corporation, Malvern, PA (http: / / www.reactionbiology.com). The substrate for HDAC-1, -2, -8 and -6 is a fluorogenic peptide derived from p53 residues 379-382 [RHKK(Ac)]. Compounds were dissolved in DMSO and tested in a 10-dose IC50 mode with 3-fold serial dilution starting at 10 pM. Trichostatin A (TSA) was the reference.

[0083] Gene expression and survival rates analysis

[0084] For the analysis of the impact on gene expression and survival rates, the TCGA- GBM database was employed to analyze the expression levels in normal brain tissue and GBM. Furthermore, the CGGA-GBM database was utilized to analyze the expression of Class I HDAC in primary and resistant GBM. The analysis of HDAC and survival rates was conducted using the CGGA database. All these data were processed through the GlioVis website (http / . / / ^l io vissbi oinfo cni o . es / ) .

[0085] Western blotting

[0086] The experimental procedure was performed as described in our previous study. On the day prior to the experiment, 5* 102cells were seeded into each well of a 6-well plate. The following day, the cells were washed twice with PBS and treated with 10 pM MPT1 A089. Theacetylation status of histone H3 and H4, DNA repair, and protein stability assay were assessed at 0-24 hours post-treatment. The antibody models and dilution ratios used are detailed in Table SI. Protein images were collected and quantified using the ChemiDoc™ Touch Imaging System (Cat# 12676029, Bio-Rad Laboratories, Inc., CA, USA).

[0087] Bioinformatics analysis

[0088] After treating PT#3R cell lines with 10 pM of MPT1 A089 for 72 hours, cells were lysed using QIAzol Lysis Reagent (Cat# 79306, QIAGEN, Hilden, Germany), and the extraction and analysis were entrusted to BioTools (Taipei, Taiwan). RNA Purity and quantification were checked using SimpliNano™ - Biochrom Spectrophotometers (Biochrom, MA, USA). RNA degradation and integrity were monitored by Qsep 100 DNA / RNA Analyzer (BiOptic Inc., Taiwan). A total amount of 1 pg total RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using a KAPA mRNA HyperPrep Kit (KAPA Biosystems, Roche, Basel, Switzerland) following the manufacturer’s recommendations, and index codes were added to assign sequences to each sample. The strand marked with dUTP was not amplified, allowing strand-specific sequencing. Finally, PCR products were purified using the KAPA Pure Beads system, and the library quality was assessed on the Qsep 100 DNA / RNA Analyzer (BiOptic Inc). The library quality was assessed on the Qubit® 2.0 Fluorometer (Thermo Scientific) and Agilent Bioanalyzer 2100 system. Finally, the library was sequenced on an Illumina NovaSeq6000 platform and 150 bp paired-end reads were generated. The raw data, generated by high-throughput sequencing (Illumina NovaSeq 6000 platform), were initially converted into raw sequenced reads using CASAVA base calling and were stored in FASTQ format. Subsequently, differential expression genes (DEGs) under two conditions were identified using standard methods. The DEGs were then subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Simultaneously, the DOSE package was employed for Disease Ontology (DO) enrichment analysis using DO, DisGeNET, and NCG databases. Additionally, gene setenrichment analysis (GSEA) was performed to identify enriched biological functions and activated pathways using a molecular signature database (MSigDB) with 1,000 permutations. MSigDB provides annotated gene sets for use with the GSEA software, including hallmark gene sets, positional gene sets, curated gene sets, motif gene sets, computational gene sets, GO gene sets, oncogenic gene sets, and immunologic gene sets.

[0089] Cell cycle

[0090] To measure the cell cycle, cells were seeded at 5* 104per well in the 12-well plate for 24h, followed by treatment with different concentrations of compounds for 72h. Next, the cells were fixed with 70% ethanol and stored at -20°C for at least 24hr. After washing, the cells were stained with 30 pg / mL of Propidium Iodide (PI, Cat# P4170, Sigma-Aldrich) and treated with 10 ng / mL of RNAseA (Cat# R6148, Sigma-Aldrich) for 30 mins, followed by detection using flow cytometry Guava® easyCyte™ Flow Cytometers and subsequent analysis using Guava ® easyCyte System 3.3 (Cytec Industries Incorporated, NJ, USA).

[0091] Homologous recombination repair assays

[0092] For the DNA homologous recombination (HR) repair assay, the following protocol was adopted. U87MG cells were co-transfected at a 2: 1 ratio using the NHEJ reporter and DsRed-Monomer plasmids. Cells were harvested 2 days after transfection and subjected to flow cytometry analysis by the Guava EasyCyte System. Only DsRed-positive cells were analyzed for HR and NHEJ efficiency to circumvent possible differences in transfection efficiencies. Data were analyzed to reveal the percentage of GFP -positive cells relative to that of DsRed- positive cells. Data were set to 1% of the background level of GFP -positive cells in every internal control.

[0093] Immunoprecipitation and DNA ubiquitination assay

[0094] The mRNA coding sequence of RAD51 (NM_001164269.2) was referenced from the National Center for Biotechnology Information. Subsequently, truncated fragments (RAD51-T1, SEQ ID NO. l; RAD51-T2, SEQ ID NO.2; RAD51-T3, SEQ ID NO.3) weredesigned based on different functional domains. The entire gene synthesis and construction into the pEGFP-Cl vector were outsourced toMDBio, Inc. (New Taipei City, Taiwan). The pEGFP- RAD51-T3 plasmid (SEQ ID NO. 4) was further modified by mutating the codons from lysine to alanine at specific positions, resulting in Mutl-Mut4 (SEQ ID Nos. 5-8). Details regarding plasmid construction are provided in Figure 6H. After overexpressing the truncated or mutated plasmids in the U87MG cell line for 24 hours, the cells were treated with MPT1A089 for an additional 24 hours. Subsequently, cells were harvested, and protein lysate was extracted using I xRIPA Lysis Buffer (Cat# 20-188, Merck Millipore). Following protein quantification, 1 pg of protein lysate was combined with 1 pg of GFP antibody and subjected to immunoprecipitation (IP) using the Catch and Release® Reversible Immunoprecipitation System (Cat# 17-500, Merck Millipore) for 12 hours. The resulting products from the IP were then subjected to immunoblotting using an anti-ubiquitination antibody.

[0095] Extraction of plasma and brain tissue samples

[0096] Following anesthesia of mice with isoflurane, blood samples were collected via cardiac puncture. The blood was centrifuged at 2000 xg for 20 minutes at 4°C to obtain plasma. Plasma samples were then mixed with an equal volume of 1 M Zinc sulfate heptahydrate solution (Cat# 221376, Sigma-Aldrich) and allowed to stand for 30 minutes. Subsequently, the mixture was centrifuged at 12000 xg for 30 minutes at 4°C, and the supernatant was collected for analysis. Brain tissues were homogenized using a Tenbroeck tissue grinder at a ratio of 5 pL DMSO per mg tissue. The homogenates were then centrifuged at 12000 xg for 30 minutes at 4°C, and the supernatant was collected for analysis.

[0097] Pharmacokinetics and brain distribution study

[0098] The compounds were prepared in DMSO at a concentration of 180 mg / mL, and TMZ at 30 mg / mL. C57BL / 6 mice were intraperitoneally injected with MPT1A089 at 360 mg / kg, TMZ at 60 mg / mL, or DMSO at 2 uL / g as the control group. Mice were sacrificed byintraperitoneal injection after 30 minutes, and plasma as well as brain tissue extracts were collected following the aforementioned procedures.

[0099] Drug inhibition of orthotopic tumor growth experiment

[0100] For orthotopic inoculation, U87MGR cells (5* 105) were suspended in 4 pL of DMEM and implanted into the brains of SCID mice. After cell implantation, mice were monitored daily for survival, and their body weight was measured every 3 days starting from the seventh day. Intraperitoneal injections were administered with DMSO (solvent control), TMZ (10 pg / kg), TMZ (10 pg / kg),+MPTl A089 (10 pg / kg or 20 pg / kg). Upon the natural death of the mice, brains were extracted and entrusted to BioTools for sectioning and HE staining.

[0101] Statistical analyses

[0102] Statistical analyses of two groups of data from western blotting, micronucleus assay, HR / NHEJ DNA repair assays, MTT, CCK8 assay, tumor weight of subcutaneously inoculated mice, RNA-seq data of TCGA-GBM dataset, qPCR, etc., were carried out using Student’s t- test with a two-tailed distribution. Multiple groups of data from the MTT assay and western blotting were analyzed using two-way analysis of variance, followed by Tukey’s multiple comparison test. The Gehan-Breslow-Wilcoxon test was used to compare the survival curves (Kaplan-Meier curve) of orthotopic GBM mice. Quantitative data (bar chart) are shown as mean ± SEM. A value ofp < 0.05 was considered statistically significant (*p < 0.05; **p < 0.01; ***p < 0.001).

[0103] The following examples are provided for illustrating the concepts of the disclosure.

[0104] Synthetic Examples 1-15

[0105] Examples 1-10 and 12-15: Synthesis of Compounds (4), (8), (11), (12), (17), (18), (19), (22), (24), (26), (31), (34), (36) and (38)

[0106] The synthesis of Compound (4) (MPT1 A089) was illustrated in Example 1, starting with the linezolid related amine (1) followed by the reductive amination with 4-formylcinnamic acid to afford the carboxylic acid (2). The carboxylic acid moiety then underwent amidecoupling reaction with THP -protected hydroxylamine to yield compound (3). Lastly, the protection group was removed under acidic conditions to afford desired Compound (4) (MPT1 A089). The synthetic routes in Examples 2-5 for preparing Compounds (8), (11), (12) and (17) differed only in the installation of the linker. Examples 6 and 7 show that the Compounds (18) and (19) were synthesized via an amide coupling reaction. The titled compounds in Examples 8-10 were synthesized with the corresponding carboxylic acid 20, and with or without substitution on the amide moiety, followed by the removal of the benzyl protection group to afford Compound (22), (24), and (26).

[0107] Reference Example 11 shows the synthesis of the starting material compound (29), starting with the linezolid related amine (1) with the acetylation of amino group to afford linezolid (27), followed by nitration with nitric acid, and then reduction of the nitro group to yield aniline 29. The further synthetic route in Examples 12-15 used compound 29 as the starting material, which then underwent similar synthetic strategies. The characterization (NMR verification and HPLC purity analysis) is also provided in each example.

[0108] Example 1 - Compound (4)

[0109] (S,E)-3-(4-((((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)amino)me thy l)pheny I) acrylic acid (2)

[0110] Suspend linezolid related amine 1 (300 mg, 1.02 mmol, 1 eq) and 4-formyl cinnamic acid (176 mg, 1.02 mmol, 1 eq) in MeOH (25 mL). The resulting reaction mixture was heated up to reflux and stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was cooled down to 0 °C, following with the addition of sodium borohydride (58 mg, 1.53 mmol, 1.5 eq) portionwise. A pale-white solid precipitated immediately, the reaction mixture was stirred for additional 2 h. After reaction finished, collected the precipitate through filtration to afford the desired product as pale-white solid (440 mg, 95 %). 'H NMR (300 MHz, DMSO-tL) 8 7.52 (dd, J= 14.9, 3.3 Hz, 1H), 7.41 (d, J= 7.9 Hz, 2H), 7.29 (d, J= 8.0 Hz, 2H), 7.22 (dd, J= 8.8, 2.5 Hz, 1H), 7.08 (dd, J= 9.4, 9.1 Hz, 1H), 7.04 (d, J= 15.9 Hz, 1H), 6.33 (d, J= 15.8 Hz, 1H), 4.78-4.70 (m, 1H), 4.07 (t, J= 8.6 Hz, 1H), 3.83-3.74 (m, 6H), 2.98 (t, J= 4.4 Hz, 4H), 2.81-2.75 (m, 2H).

[0111] (E)-3-( 4-( ((( (S)-3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)a- mino)methyl)phenyl)-N-( tetrahydro-2H-pyran-2-yl)oxy)acrylamide (3)

[0112] Suspend compound 2 (400 mg, 0.88 mmol, 1 eq), HOBt (144 mg, 1.05 mmol, 1.2 eq), EDCi (254 mg, 1.32 mmol, 1.5 eq) in dry DMF (5 mL) then NMM ( 0.14ml, 1.32 mmol, 1.5 eq) was added. The reaction mixture was stirred for 10 min, then added <9-(tetrahydro-27 / - pyran-2-yl) hydroxylamine (124 mg, 1.05 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCh and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH=15: 1) to afford the desired product as pale-white solid (340 mg, 70 %).XH NMR (300 MHz, DMSO-tL) 5 11.19 (br s, 1H), 7.52-7.43 (m, 4H), 7.36 (d, J= 8.16 Hz, 2H), 7.19 (dd, J= 8.4, 1.9 Hz, 1H), 7.04 (dd, J= 9.6, 9.2 Hz, 1H), 6.47 (d, J= 16.1 Hz, 1H), 4.89 (s, 1H), 4.71 (m, 1H), 4.04 (t, J= 8.91 Hz, 1H), 3.95 (m, 1H), 3.80-3.71 (m, 3H), 3.72 (t,J= 4.5 Hz, 4H), 3.48-3.56 (m, 1H), 2.95 (t, J= 4.7 Hz, 4H), 2.77 (d, J= 5.19 Hz, 2H), 1.68 (br s, 3H), 1.52 (br s, 3H).

[0113] (S,E)-3-(4-((((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)ami- no)methyl)phenyl)-N-hydr oxyacrylamide (4) (the titled compound)

[0114] Dissolved 3 (340 mg, 0.62 mmol, 1 eq) in 5 % TFA in MeOH (10 mL) and stirred for 16 h at room temperature. The reaction is check by TLC. After the reaction finished, solvent was removed by reduced pressure. The residue was triturated with a mixture of diethyl ether and methanol, filtered, dried under vacuum to afford the desired product as pale-white solid. (290 mg, quant.); HPLC purity=95.14%; m.p.=162.2 °C; 'H NMR (300 MHz, DMSO-cf.) 8 10.80 (br s, 1H), 9.32 (br s, 1H), 9.11 (br s, 1H)7.63 (d, J= 8.2, 2H), 7.53 (d, J= 8.3 Hz, 2H), 7.52 (d, J= 14.9 Hz, 2H), 7.46 (dd, J= 14.9, 2.4 Hz, 1H), 7.17 (dd, J= 8.9, 2.1 Hz, 1H), 7.07 (dd, J= 9.3, 9.0 Hz, 1H), 6.50 (d, J= 15.8 Hz, 1H), 5.04-4.90 (m, 1H), 4.24 (br s, 2H), 4.15 (t, J= 9.3 Hz, 1H), 3.80 (dd, J= 9.3, 6.6 Hz, 1H), 3.72 (t, J= 4.8 Hz, 4H), 2.95 (t, J= 4.7 Hz, 4H).13C NMR (150 MHz, DMSO-t / e) 6 158.85, 158.23, 155.77, 153.93, 136.27, 133.51, 131.06, 128.10, 120.41, 119.70, 118.66, 116.68, 114.84, 107.41, 69.38, 66.57, 51.11, 51.09, 50.83, 49.42, 49.02, 47.93. HRMS calcd for C24H27FN4O5 [M + H]+471.2038, found 471.2044.

[0115] Example 2 - Compound (8)

[0116] Reagents and conditions :(a) 1. monomethyl suberate, oxalyl chloride, DCM, 0 °C to r.t., 2h 2. pyridine, DCM, 0 °C to r.t., 16h (b) IN LiOH(aq), dioxane, r.t., 16 h (c) NH2OTHP, EDCi, HOBt, NMM, DMF, r.t., 16 h (d) 5 % TFA(MeoH), r.t., 16 h.

[0117] methyl (S)-8-(((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)a- mino)-8-oxooctanoate (5)

[0118] Dissolve monomethyl suberate (192 mg, 1.02 mmol, 1 eq) in dry DCM (3 mL), then added oxalyl chloride (0.13 mL, 1.02 mmol, 1 eq) under 0 °C. The resulting reaction mixture was stirred for 2 h at room temperature under N2 atmosphere. After reaction finished, solvent and excess oxalyl chloride was removed under reduced pressure, the crude product was used in next step without further purification.

[0119] The linezolid related amine 1 (300 mg, 1.02 mmol, 1 eq) was taken up in dry DCM (3 mL) added dropwise into crude acyl chloride, followed by pyridine (0.12 mL, 1.53 mmol, 1.5 eq) under 0 °C. The reaction mixture was warmed up to ambient temperature stirred for 16 h under N2 atmosphere. The reaction was check by TLC. After reaction finished, the reaction mixture was washed with water three times and brine once. The organic layer was collected, dried over MgSCU and removed solvent under reduced pressure to get the crude product. Thecrude product was purified by column chromatography (DCM:MeOH=15: l) to afford the desired product as pale yellow solid (200 mg, 42 %).1H NMR (300 MHz, CDCh) 8 7.45 (dd, J= 14.5, 2.6 Hz, 1H), 7.07 (dd, J= 8.9, 2.6 Hz, 1H), 6.96 (dd, J= 8.9, 8.8 Hz, 1H), 6.02 (dd, J= 6.2, 5.8 Hz, 1H), 4.71-4.79 (m, 1H), 4.01 (t, J= 9.0 Hz, 1H), 3.87 (t, J= 4.7 Hz, 4H), 3.75 (dd, J= 9.1, 6.5 Hz, 1H), 3.67-3.63 (m, 2H), 3.65 (s, 3H), 3.06 (t, .7= 4,6 Hz, 4H), 2.36-2.16 (m, 4H), 1.63-1.54 (m, 2H), 1.36-1.33(m, 2H), 1.30-1.25 (m, 4H).

[0120] (S)-8-(((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)amino)-8- oxooctanoic acid (6)

[0121] To a solution of 5 (500 mg, 1.08 mmol, 1 eq) in dioxane(18 mL) ,1N LiOH(aq) (1.07 mL, 2.7 mmol, 2.5 eq) was added. The resulting reaction mixture was stirred for 16 h at room temperature. The reaction was checked by TLC. After starting material fully transformed to lithium salt, dioxane was removed under reduced pressure. Then adjusted the reaction mixture to pH<2 by using 3N HCl(aq), white solid precipitated. Filtered, collected the precipitate, dried under vacuum to afford the desired product as white solid (310 mg, 64 %).1H NMR (300 MHz, DMSO-tL) 6 8.19 (t, J= 6 Hz, 1H), 7.50 (dd, J= 15.1, 2.6 Hz, 1H), 7.18 (dd, J= 8.9, 2.6 Hz, 1H), 7.07 (dd, J= 9.3, 9.0 Hz, 1H), 4.78-4.69 (m, 1H), 4.09 (t, J= 92 Hz, 1H), 3.75 (t, J= 4.6 Hz, 4H), 3.72-3.69 (m, 1H), 3.54-3.38 (m, 2H), 2.97 (t, J= 4.7 Hz, 4H), 2.27- 2.06 (m, 4H), 1.46-1.42 (m, 4H), 1.25-1.17 (m, 4H).

[0122] N1-( ((S)-3-( 3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)-N8-((tetrahydro-2H-pyran-2-yl)oxy)octanediamide (7)

[0123] Suspend 6 (260 mg, 0.58 mmol, 1 eq), HOBt (93 mg, 0.69 mmol, 1.2 eq), EDCi (165 mg, 0.86 mmol, 1.5 eq) in dry DMF (5 mL) then NMM ( 0.1ml, 0.86 mmol, 1.5 eq) was added. The reaction mixture was stirred for 10 min, then added <9-(tetrahydro-2 / / -pyran-2-yl) hydroxylamine (118 mg, 0.69 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once.The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH=15: l) to afford the desired product as yellow solid (320 mg, quant.) 'H NMR (300 MHz, DMSO-t / e) 8 10.88 (s, 1H), 8.19 (t, J= 6.2 Hz, 1H), 7.50 (dd, J= 15.1, 2.5 Hz, 1H), 7.18 (dd, J= 9.1, 2.7 Hz, 1H), 7.08 (dd, J= 9.3, 9.1 Hz, 1H), 4.81 (br s, 1H), 4.78-4.70 (m, 1H), 4.10 (t, J= 9.0 Hz, 1H), 3.93-3.89 (m, 1H), 3.75 (t, J= 4.4 Hz, 4H), 3.72-3.69 (m, 1H), 3.53- 3.38 (m, 3H), 2.98 (t, J= 4.7 Hz, 4H), 2.11 (t, J= 7.3 Hz, 2H), 1.95 (t, J= 7.4 Hz, 2H), 1.66-1.43 (m, 10H), 1.18 (m, 4H).

[0124] (S -N1-( 3-( 3-fhioro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)-N8- hydroxyoctanediamide (8) (the titled compound)

[0125] Dissolved 7 (320 mg, 0.58 mmol, 1 eq) in 5 % TFA in MeOH (10 mL) and stirred for 16 h at room temperature. The reaction is check by TLC. After the reaction finished, solvent was removed by reduced pressure. The crude product was purified by column chromatography (DCM:MeOH=15: l) to afford the desired product as orange solid (190 mg, 70 %); HPLC purity=99.12%; m.p.=136.0 °C;XH NMR (300 MHz, DMSO-t / e) 6 10.30 (br s, 1H), 8.63 (br s, 1H), 8.17 (t, J= 5.9 Hz, 1H), 7.48 (dd, J= 15.1, 2.5 Hz, 1H), 7.16 (dd, J= 8.7, 2.3 Hz, 1H), 7.05 (dd, J= 9.3, 9.1 Hz, 1H), 4.74-4.68 (m, 1H), 4.07 (t, J= 8.9 Hz, 1H), 3.73 (t, J= 4.5 Hz, 4H), 3.70-3.68 (m, 1H), 3.50-3.38 (m, 2H), 2.95 (t, J= 4.6 Hz, 4H), 2.06 (t, J= 7.4 Hz, 2H), 1.89 (t, J= 7.4 Hz, 2H), 1.40 (br s, 4H), 1.15 (br s, 4H).13C NMR (150 MHz, DMSO-t / e) 6 173.53, 169.53, 155.83, 154.50, 135.96, 133.90, 119.70, 114.45, 107.09, 71.99, 66.59, 51.15, 49.03, 47.65, 41.70, 35.67, 32.65, 21.11, 28.77, 25.64. HRMS calcd for C22H32FN4O6 [M + H]+467.2300, found 467.2307.

[0126] Examples 3 and 4 - Compounds (11) and (12)

[0127] Reagents and conditions: (a) 1. 4-formyl benzoic acid, MeOH, reflux, 16 h 2. NaBH4, 0 °C to r.t., 2 h (b) NH2OTHP, EDCi, HOBt, NMM, DMF, r.t., 16 h (c) 5 % TFA(MeoH), r.t., 16 h (d) o-phenylenediamine, EDCi, HOBt, NMM, DMF, r.t., 16 h.

[0128] (S)-2-(4-((((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5- yl)methyl)amino)methyl)phenyl)benzoic acid (9)

[0129] Suspend linezolid related amine 1 (500 mg, 1.69 mmol, 1 eq) and 4-formylbenzoic acid (254 mg, 1.69 mmol, 1 eq) in MeOH (35 mL). The resulting reaction mixture was heated up to reflux and stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After starting material vanished, the reaction mixture was cooled down to 0 °C, added sodium borohydride (58 mg, 1.53 mmol, 1.5 eq), and stirred for additional 2 h. After reaction finished, the solvent was removed under reduced pressure to afford yellow solid (1.13 g, crude). The crude product was used in next step without further purification.

[0130] 4-(((((S)-3-(3-fhioro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)amin- o)methyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (10)

[0131] Suspend crude product 9 (1.13 g), HOBt (273 mg, 2.02 mmol, 1.2 eq), EDCi (487 mg, 2.54 mmol, 1.5 eq) in dry DMF (10 mL) then NMM ( 0.28ml, 2.54 mmol, 1.5 eq) was added. The resulting reaction mixture was stirred for 10 min, then added <9-(tetrahydro-27 / - pyran-2-yl) hydroxylamine (237 mg, 2.02 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSC and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH=15: 1) to afford the desired product as pale- white solid (720 mg, 80 %). 'H NMR (300 MHz, DMSO-tL) 8 11.61 (s, 1H), 7.74 (d, J= 8.2 Hz, 2H), 7.53 (dd, J= 14.9, 2.4 Hz, 1H), 7.44 (d, J= 8.2 Hz, 2H), 7.22 (dd, J= 8.4, 1.7 Hz, 1H), 7.08 (dd, J= 9.2, 9.0 Hz, 1H), 5.01 (br s, 1H), 4.79-4.70 (m, 1H), 4.08 (t, J= 8.9 Hz, 2H), 3.82 (t, J= 4.8 Hz, 3H), 3.76 (t, J= 4.9 Hz, 4H), 3.55-3.49 (m, 2H), 2.98 (t, J= 4.6 Hz, 1H), 2.80 (d, J= 5.6 Hz, 1H), 1.74 (br s, 3H), 1.57 (br s, 3H).

[0132] 5- VI-1.10 (S)-4-((((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidm-5- yl)methyl)amin-o)methyl)-N-hydroxybenzamide (11) (the titled compound)

[0133] Dissolved 10 (720 mg, 1.36 mmol, 1 eq) in 5 % TFA in MeOH (10 mL) and stirred for 16 h at room temperature. The reaction is check by TLC. After the reaction finished, solvent was removed by reduced pressure. The residue was triturated with a mixture of ethyl acetate and methanol, filtered, dried under vacuum to afford the desired product as pale-white solid (380 mg, 63 %); HPLC purity=96.55%; m.p =194.5 °C;1HNMR (300 MHz, DMSO-t / e) 6 11.29 (br s, 1H), 9.30 (br s, 1H), 9.09 (br s, 1H), 7.81 (d, J= 8.2 Hz, 2H), 7.57 (d, J= 8.2 Hz, 2H), 7.47 (dd, J= 15.0, 2.5 Hz, 1H), 7.18 (dd, J= 9.2, 2.3 Hz, 1H), 7.07 (dd, J= 9.3, 9.2 Hz, 1H), 5.04- 4.99 (m, 1H), 4.26 (br s, 2H), 4.16 (t, J= 14.9, 2.4 Hz, 1H), 3.80 (dd, J= 9.2, 7.9 Hz, 1H), 3.73(t, J= 4.8 Hz, 4H), 2.96 (t, J= 4.7 Hz, 4H).13C NMR (150 MHz, DMSO-cf.) 6 155.77, 154.15, 153.87, 136.28, 133.48, 130.59, 127.58, 119.70, 114.83, 107.41, 69.15, 66.57, 51.11, 50.64, 49.46, 47.88. HRMS calcd for C24H27FN4O5 [M + H]+445.1882, found 445.1887.

[0134] (S)-N-(2-aminophenyl)-4-((((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5- yl)methyl)amino)methyl)benzamide (12) (the title compound)

[0135] Suspend crude product 9 (670 mg), HOBt (162 mg, 1.21 mmol, 1.2 eq), EDCi (288 mg, 1.52 mmol, 1.5 eq) in dry DMF (10 mL) then NMM ( 0.28ml, 2.54 mmol, 1.5 eq) was added. The resulting reaction mixture was stirred for 10 min, then added o-phenylenediamine (130 mg, 1.21 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (EA) to afford the desired product as yellow solid (130 mg, 25 %); HPLC purity=95.07%; m.p.=136.4 °C1H NMR (300 MHz, DMSO-t / e) 8 9.61 (s, 1H), 7.93 (d, J= 8.2 Hz, 2H), 7.51 (dd, J=15.2, 2.6 Hz, 1H), 7.46 (d, .7= 8.3 Hz, 2H), 7.21 (ddd, J= 8.9, 2.5, 0.6 Hz, 1H), 7.16 (dd, J= 7.9, 1.4 Hz, 1H), 7.06 (dd, J= 10.0, 9.0 Hz, 1H), 6.97 (ddt, J= 7.9, 1.5, 0.6 Hz, 1H).13C NMR (75 MHz, DMSO-t / e) 6 170.55, 165.64, 156.69, 147.70, 144.45, 143.60, 142.24, 140.30, 133.51, 128.31, 127.40, 127.15, 126.92, 123.85, 116.74, 116.61, 115.98, 101.83, 72.93, 66.57, 55.38, 50.79, 49.42, 46.32, 42.05, 22.96. HRMS calcd for C28H31FN5O4 [M + H]+520.2355, found 520.2351.

[0136] Example 5 - Compound (17)

[0137] Reagents and conditions: (a) 3 -bromobenzenesulfonyl chloride, pyridine, DCM, 0 °C to r.t., 1 h (b) methyl acrylate, Pd(OAc)2, PPh3, NaHCO3, Et3N, DMF, 120 °C, 16 h (c) IN LiOH(aq), dioxane, r.t., 16 h (d) NH2OTHP, EDCi, HOBt, NMM, DMF, r.t., 16 h (e) 5 % TFA(MeOH), r.t., 16 h.

[0138] (R)-3 -bromo-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methy- l)benzeneesulfonamide (13)

[0139] To a solution of linezolid related amine 1 (600 mg, 2.04 mmol, 1 eq), pyridine (0.24 ml, 3.06 mmol, 1.5 eq) in dry DCM (20 mL), 3 -bromobenzenesulfonyl chloride (0.36 mL, 2.44 mmol, 1.2 eq) was added dropwise at 0 °C. Then warmed up to room temperature, the reaction mixture was stirred for 2 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with brine once. The organic layer was collected, dried over MgSC and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (EA:n-Hexane = 1 : 1) to afford the desired product as white solid (550 mg, 52 %). 'H NMR (300 MHz, DMSO-tL) 8 8.27 (br s, 1H), 7.95 (t, J= 1.8 Hz, 1H), 7.86 (ddd, J= 7.8, 1.9, 0.8 Hz, 1H), 7.81 (ddd, J= 7.8, 2.0, 1.1 Hz, 1H), 7.56 (t, J= 8.0 Hz, 1H), 7.47 (dd, J= 15.2, 2.6 Hz, 1H), 7.15 (dd, J= 8.9, 2.6 Hz, 1H), 7.06 (dd, J= 9.2, 8.9 Hz, 1H), 4.74-4.66 (m,1H), 4.07 (t, J= 9.2 Hz, 1H), 3.76-3.70 (m, 5H), 3.15 (dd, J= 6.8, 5.4 Hz, 2H), 2.96 (t, .7=4,6Hz, 4H).

[0140] methyl (R,E)-3-(3-(N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5- yl)methyl)sulfamoyl)phenyl)acrylate (14)

[0141] Suspend 13 (550 mg, 1.07 mmol, 1 eq), Pd(0Ac)2 (22 mg, 0.1 mmol, 0.1 eq), PPF13 (32 mg, 0.2 mmol, 0.2 eq), NaHCCh (90 mg, 1.07 mmol, leq) in dry DMF (10 ml), then added methyl acrylate (0.11 mL, 1.28 mmol, 1.2 eq), and EtsN (0.17 mL, 1.07 mmol, 1 eq). The resulting reaction mixture was heated up to 120 °C, stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was filtered through celite cake, washed with EA and collected the filtrate. The solvent was removed under reduced pressure and the residue was purified by flash column chromatography (DCM:MeOH = 15:1) to afford the product with slice methyl acrylate as brown solid (630 mg, crude). 'H NMR (300 MHz, DMSO-tL) 8 8.18 (br s, 1H), 8.13 (t, J= 1.6 Hz, 1H), 8.03 (d, J= 7.9 Hz, 1H), 7.86 (ddd, J= 7.8, 1.9, 1.1 Hz, 1H), 7.76 (d, J= 16.1 Hz, 1H), 7.67 (dd, J= 7.9, 7.8 Hz, 1H), 7.49 (dd, J= 15.1, 2.5 Hz, 1H), 7.16 (dd, J= 8.9, 2.6 Hz, 1H), 7.08 (dd, J= 9.2, 9.0 Hz, 1H), 6.77 (d, J= 16.1 Hz, 1H), 4.78-4.69 (m, 1H), 4.09 (t, J= 9.1 Hz, 1H), 3.77 (s, 3H), 3.77-3.73 (m, 5H), 3.21-3.17 (m, 2H), 2.98 (t, J= 4.7 Hz, 4H).

[0142] (R,E)-3-(3-(N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)su- lfamoyl)phenyl)acrylic acid (15)

[0143] To a solution of crude 14 (630 mg) in dioxane(20 mL), IN LiOH(aq) (3.13 mL, 3.13 mmol, 2.5 eq) was added. The resulting reaction mixture was stirred for 16 h at room temperature. The reaction was checked by TLC. After starting material fully transformed to lithium salt, dioxane was removed under reduced pressure. Then adjusted the reaction mixture to pH<2 by using 3N HCl(aq), white solid precipitated. Filtered, collected the precipitate, dried under vacuum to afford the desired product as yellow solid (240 mg, 44 %). 'H NMR (300 MHz, DMSO-tL) 6 8.23 (t, J= 6.1 Hz, 1H), 8.01 (s, 1H), 7.96 (d, J= 7.6 Hz, 1H), 7.82 (d, J=7.7 Hz, 1H), 7.64 (d, J= 16.0 Hz, 1H), 7.63 (t, J= 7.6 Hz, 1H), 7.47 (dd, J= 15.2, 2.4 Hz, 1H), 7.14 (dd, J= 8.8, 2.2 Hz, 1H), 7.08 (dd, J= 9.1, 9.0 Hz, 1H), 6.63 (d, J= 16.1 Hz, 1H), 4.73-4.68 (m, 1H), 4.06 (t, J= 9.2 Hz, 1H), 3.78-3.72 (m, 1H), 3.74 (t, J= 4.8 Hz, 4H), 3.18-3.12 (m, 2H), 2.97 (t, J= 4.6 Hz, 1H).

[0144] (E)-3-(3-(N-(((R)-3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)meth- yl)sulfamoyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (16)

[0145] Suspend 15 (240 mg, 0.47 mmol, 1 eq), HOBt (76 mg, 0.56 mmol, 1.2 eq), EDCi (136 mg, 0.71 mmol, 1.5 eq) in dry DMF (4 mL) then NMM (0.08 ml, 0.71 mmol, 1.5 eq) was added. The reaction mixture was stirred for 10 min, then added <9-(tetrahydro-27 / -pyran-2-yl) hydroxylamine (66 mg, 0.56 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH = 15: 1) to afford the desired product as yellow solid (240 mg, 84 %). 'HNMR (300 MHz, DMSO-t / e) 8 8.00 (s, 1H), 7.78-7.85 (m, 2H), 7.64 (d, J= 153 Hz, 1H), 7.52-7.64 (m, 1H), 7.47 (dd, J= 15.0, 2.4 Hz, 1H), 7.15 (dd, J= 8.8, 2.3 Hz, 1H), 7.06 (dd, J= 9.2, 8.9 Hz, 1H), 6.63 (d, J= 15.7 Hz, 1H), 4.93 (br s, 1H), 4.73-4.67 (m, 1H), 4.07 (t, J= 9.5 Hz, 1H), 3.99- 3.90 (m, 1H), 3.77-3.72 (m, 1H), 3.74 (t, J= 4.9 Hz, 4H), 3.54 (d, J= 12.4 Hz, 1H), 3.18-3.14 (m, 2H), 2.97 (t, J= 4.9 Hz, 4H), 1.70 (br s, 3H), 1.55 (br s, 1H).

[0146] (R,E)-3-(3-(N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)su- lfamoyl)phenyl)-N-hydr oxyacrylamide (17) (the titled compound)

[0147] Dissolved 16 (240 mg, 0.40 mmol, 1 eq) in 5 % TFA in MeOH (10 mL) and stirred for 16 h at room temperature. The reaction is check by TLC. After the reaction finished, solvent was removed by reduced pressure. The residue was triturated with a mixture of ethyl acetate and methanol, filtered, dried under vacuum to afford the desired product as orange solid (170mg, 82 %); HPLC purity=95.07%; m.p =123.0°C; 'HNMR (300 MHz, DMSO-cf.) 6 8.17 (t, J= 6.2 Hz, 1H), 7,97 (s, 1H), 7.80 (dd, J= 9.3, 8.8 Hz, 2H), 7.62 (t, J= 7.8 Hz, 1H), 7.51 (d, J= 15.1 Hz, 1H), 7.46 (dd, J= 15.1, 2.4 Hz, 1H), 7.14 (dd, J= 8.9, 2.3 Hz, 1H), 7.05 (dd, J= 9.2, 9.0 Hz, 1H), 6.56 (d, J= 15.1 Hz, 1H), 4.74-4.65 (m, 1H), 4.06 (t, J= 9.1 Hz, 1H), 3.76-3.71 (m, 1H), 3.72 (t, J= 5.0 Hz, 1H), 3.16-3.10 (m, 2H), 2.95 (t, J= 4.7 Hz, 4H).13C NMR (75 MHz, DMSO-t / e) 5 156.67, 154.38, 153.44, 141.62, 136.44, 136.08, 135.96, 133.89, 133.74, 132.14, 130.53, 127.43, 125.14, 119.77, 114.53, 107.24, 106.90, 77.67, 66.63, 60.23, 51.20, 46.35, 45.65, 21.23, 14.56. HRMS calcd for C23H26FN4O7S [M + H]+521.1501, found 521.1508.

[0148] Examples 6 and 7 - Compounds (18) and (19)MPT1A095 (19)

[0149] Reagents and conditions: (a) 3,4,5-trimethoxybenzoyl chloride, pyridine, DCM, 0 °C to r.t., 16 h (b) 5-chlorosalicylic acid, EDCi, HOBt, NMM, DMF, r.t, 16 h.

[0150] (S)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)-3,4,5- trimethoxybenzamide (18) (the titled compound)

[0151] To a solution of linezolid related amine 1 (300 mg, 1.02 mmol, 1 eq), pyridine (0.13 ml, 1.33 mmol, 1.3 eq) in dry DCM (5.7 mL), 3,4,5-trimethoxybenzoyl chloride (282 mL, 1.22 mmol, 1.2 eq) was added dropwise at 0 °C. The resulting reaction mixture was warmed up to room temperature stirred for 2 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with brine once. The organic layer was collected, dried over MgSC and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (EA:MeOH = 1 : 1) to afford the desired product as white solid (250 mg, 52 %); HPLC purity=99.89%; m.p =121.3; 'HNMR (300 MHz, DMSO-t / e) 8 8.81 (t, J= 5.9 Hz, 1H), 7.50 (dd, J= 15.0, 2.5 Hz, 1H), 7.21 (dd, J= 8.8, 2.3 Hz, 1H), 7.18 (s, 2H), 7.07 (dd, J= 9.4, 9.0 Hz, 1H), 4.91-4.80 (m, 1H), 4.17 (t, J= 9.1 Hz, 1H), 3.86 (dd, J= 9.2, 7.5 Hz, 1H), 3.82 (s, 6H), 3.75 (t, J= 4.8 Hz, 4H), 3.71 (s, 3H), 3.64 (t, J= 5.5 Hz, 2H), 2.97 (t, J= 4.6 Hz, 1H).13C NMR (75 MHz, DMSO-t / e) 6 166.95, 156.67, 154.59, 153.03, 140.57, 136.08, 135.96, 133.96, 133.82, 129.64, 119.73, 114.61, 107.30, 106.96, 105.37, 71.97, 66.63, 60.55, 56.43, 51.20, 47.99, 42.93. HRMS calcd for C24H29FN3O7 [M + H]+490.1984, found 490.1991.

[0152] (S)-5-chloro-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)- 2-hydroxybenzamide (19) (the titled compound)

[0153] Suspend 5-chlorosalicylic acid (211 mg, 1.22 mmol, 1.2 eq),m HOBt (165 mg, 1.22 mmol, 1.2 eq), EDCi (293 mg, 1.53 mmol, 1.5 eq) in dry DMF (4 mL) then NMM (0.13 ml, 1.22 mmol, 1.5 eq) was added. The reaction mixture was stirred for 10 min, then added linezolid related amine 1 (300 mg, 1.02 mmol, 1 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (EA:n- Hexane = 1 :2) to afford the desired product as yellow solid (230 mg, 50 %); HPLCpurity=99.69%; m.p.=239.0 °C; *H NMR (300 MHz, DMSO ) 8 12.22 (br s, 1H), 9.11 (t, J= 5.0 Hz, 1H), 7.91 (d, J= 2.7 Hz, 1H), 7.49 (dd, J= 15.1, 2.5 Hz, 1H), 7.45 (dd, J= 8.9, 2.6 Hz, 1H), 7.20 (dd, J= 8.6, 2.0 Hz, 1H), 7.07 (dd, J= 9.3, 9.0 Hz, 1H), 6.97 (d, J= 8.9 Hz, 1H), 4.93- 4.84 (m, 1H), 4.17 (t, J= 9.0 Hz, 1H), 3.84 (dd, J= 9.0, 7.6 Hz, 1H), 3.75 (t, J= 4.8 Hz, 4H), 3.69 (t, J= 5.6 Hz, 2H), 2.97 (t, J= 4.7 Hz, 1H).13C NMR (75 MHz, DMSO-t / e) 6 168.04, 158.38, 156.67, 154.47, 153.44, 136.12, 133.91, 128.32, 122.96, 119.70, 117.78, 114.65, 107.38, 107.03, 71.60, 66.63, 51.19, 48.00, 42.53. HRMS calcd for C21H22CIFN3O5 [M + H]+450.1227, found 450.1227.

[0154] Examples 8-10 - Compounds (22), (24) and (26)

[0155] Reagents and conditions: (a) EDCi, HOBt, NMM, DMF, r.t., 16 h (b) H2, Pd / C, MeOH, r.t., 16 h (c) Mel or EtI, NaH, DMF, 0 °C to r.t., 8 h.

[0156] (S)-2, 4-bis(benzyloxy)-N-( (3-( 3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5- yl )methyl) -5-isopropylbenzamide (21)

[0157] Suspend 20 (459 mg, 1.22 mmol, 1.2 eq),m HOBt (165 mg, 1.22 mmol, 1.2 eq), EDCi (293 mg, 1.53 mmol, 1.5 eq) in dry DMF (4 mL) then NMM (0.13 ml, 1.22 mmol, 1.5 eq) was added. The reaction mixture was stirred for 10 min, then added linezolid related amine1 (300 mg, 1.02 mmol, 1 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM) to afford the desired product as white solid (350 mg, 53 %).JH NMR (300 MHz, DMSO-tL) 5 8.36 (t, J= 5.8 Hz, 1H), 7.63 (s, 1H), 7.50-7.30 (m, 11H), 7.16 (dd, J= 8.7, 3.2 Hz, 1H), 7.04 (dd, J= 9.3, 9.1 Hz, 1H), 6.88 (s, 1H), 5.28 (s, 2H), 5.17 (s, 2H), 4.83-4.77 (m, 1H), 4.07 (t, J= 9.21 Hz, 1H), 3.80-3.70 (m, 1H), 3.73 (t, J= 5.0 Hz, 4H), 3.63 (q, J= 5.4 Hz, 2H), 3.21-3.12 (m, 1H), 2.95 (t, J= 4.5 Hz, 4H), 1.11 (d, J= 6.9 Hz, 6H).

[0158] (S)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)-2, 4- dihydroxy-5-isopropylbenzamide (22) (the titled compound)

[0159] Dissolve 21 (200 mg, 0.31 mmol, 1 eq) in MeOH (10 mL), then 5 % Pd / C (cat.) was added. After three times degas, the resulting reaction mixture was stirred for 16 h under H2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was filtered through celite cake, washed with EA and collected the filtrate. The solvent was removed under reduced pressure and the residue was purified by column chromatography (DCM:MeOH = 15: 1) to afford the desired product as colorless solid (150 mg, quant.); HPLC purity=98.98%; m.p =113.8 °C;XH NMR (300 MHz, DMSO ) 8 7.56 (s, 1H), 7.48 (dd, J= 15.0, 2.5 Hz, 1H), 7.18 (dd, J= 9.0, 2.1 Hz, 1H), 7.04 (dd, J= 9.3, 8.9 Hz, 1H), 6.26 (s, 1H), 4.87-4.80 (m, 1H), 4.15 (t, J= 8.9 Hz, 1H), 3.81 (dd, J= 9.1, 7.6 Hz, 1H), 3.73 (t, J= 4.8 Hz, 4H), 3.64-3.58 (m, 2H), 3.12-3.01 (m, 1H), 2.95 (t, J= 4.7 Hz, 4H), 1.12 (d, J= 6.9 Hz, 6H).13C NMR (75 MHz, DMSO-t / e) 170.36, 160.28, 154.53, 136.11, 133.94, 126.67, 125.97, 119.67, 114.66, 107.03, 106.68, 102.87, 71.85, 66.63, 51.20, 48.00, 42.41, 26.43, 23.16. HRMS calcd for C24H29FN3O6 [M + H]+474.2035, found 474.2045.

[0160] (S)-2,4-bis(benzyloxy)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5- yl)methyl)-5-isopropyl-N-methylbenzamide (23)

[0161] Suspend 21 (300 mg, 0.46 mmol, 1 eq), 60 % NaH in mineral oil (24 mg, 0.6 mmol, 1.3 eq) in dry DMF (3 mL). Then Mel (0.03 mL, 0.6 mmol, 1.3 eq) was added dropwise at 0 °C. The resulting reaction mixture was warmed up to room temperature stirred for 8 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH = 30: 1) to afford the desired product as colorless solid (300 mg, 98 %).JH NMR (300 MHz, CDCI3) 8 7.40 (dd, J= 14.6, 2.6 Hz, 1H), 7.38-7.20 (m, 10H), 7.04 (dd, J= 8.6, 2.1 Hz, 1H), 7.03 (s, 1H), 6.90-6.84 (m, 1H), 6.45 (s, 1H), 5.02-4.98 (m, 1H), 4.98 (s, 2H), 4.92 (s, 2H), 3.92-3.82 (m, 2H), 3.84 (t, J= 4.7 Hz, 4H), 3.30-3.17 (m, 2H), 3.04-2.98 (m, 5H), 3.01 (s, 3H), 1.15 (d, J= 6.9 Hz, 6H)

[0162] (S)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)-2, 4- dihydroxy-5-isopropyl-N-methylbenzamide (24) (the titled compound)

[0163] Dissolve 23 (300 mg, 0.45 mmol, 1 eq) in MeOH (10 mL), then 5 % Pd / C (cat.) was added. After three times degas, the resulting reaction mixture was stirred for 16 h under H2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was filtered through celite cake, washed with EA and collected the filtrate. The solvent was removed under reduced pressure and the residue was purified by column chromatography (DCM:MeOH = 60: 1) to afford the desired product as colorless solid (100 mg, 47 %); HPLC purity=99.56%; m.p =118.3 °C;XH NMR (300 MHz, DMSO-tL) 69.50-9.46 (m, 2H), 7.50 (dd, J= 15.5, 2.1 Hz, 1H), 7.20 (dd, J= 9.2, 2.5 Hz, 1H), 7.08 (dd, J= 9.3, 9.2 Hz, 1H), 6.78 (s, 1H), 6.36 (s, 1H), 4.90-5.01 (m, 1H), 4.11 (t, J= 9.3 Hz, 1H), 3.92-3.62 (m, 3H), 3.76 (t, J= 4.9 Hz, 4H), 3.10-2.99 (m, 1H), 2.98 (t, J= 4.6 Hz, 4H), 2.98 (s, 3H), 1.08 (d, J= 6.9 Hz, 6H).13C NMR(150 MHz, DMSO-tL) 6 156.52, 155.84, 154.37, 136.03, 133.83, 125.86, 119.71, 114.75, 107.19, 102.61, 66.59, 51.15, 49.03, 47.86, 31.12, 26.23, 23.03. HRMS calcd for C25H31FN3O6 [M + H]+488.2191, found 488.2197.

[0164] (S)-2,4-bis(benzyloxy)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5- yl)methyl)-5-isopropyl-N-ethylbenzamide (25)

[0165] Suspend 21 (250 mg, 0.38 mmol, 1 eq), 60 % NaH in mineral oil (20 mg, 0.6 mmol, 1.3 eq) in dry DMF (3 mL). Then EtI (0.04 mL, 0.5 mmol, 1.3 eq) was added dropwise at 0 °C. The resulting reaction mixture was warmed up to room temperature stirred for 8 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH = 30: 1) to afford the desired product as colorless solid (240 mg, 93 %).1H NMR (300 MHz, CDCI3) 8 7.45 (dd, J= 14.6, 2.4 Hz, 1H), 7.48-7.24 (m, 10H), 7.15-7.05 (m, 1H), 7.08 (s, 1H) , 6.99-6.90 (m, 1H), 6.57-6.61 (m, 1H), 5.10-4.93 (m, 5H), 4.17-1.07 (m, 1H), 4.02-3.77 (m, 3H), 3.90 (q, J= 4.2 Hz, 4H), 3.47-3.28 (m, 1H), 3.33 (q, J= 7.1 Hz, 2H), 3.13- 2.98 (m, 4H), 1.22 (d, J= 6.8 Hz, 6H), 1.08 (t, J= 7.1 Hz, 3H).

[0166] (S)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)-2, 4- dihydroxy-5-isopropyl-N-ethylbenzamide (26) (the titled compound)

[0167] Dissolve 25 (240 mg, 0.35 mmol, 1 eq) in MeOH (10 mL), then 5 % Pd / C (cat.) was added. After three times degas, the resulting reaction mixture was stirred for 16 h under H2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was filtered through celite cake, washed with EA and collected the filtrate. The solvent was removed under reduced pressure and the residue was purified by column chromatography (DCM:MeOH = 60: 1) to afford the desired product as colorless solid (160 mg, 91 %); HPLC purity=96.98%; m.p =121.0 °C; *HNMR (300 MHz, DMSO-tL) 6 9.44 (br s, 2H), 7.50 (dd, J=15.4, 2.6 Hz, 1H), 7.20 (dd, J= 9.5, 2.6 Hz, 1H), 7.08 (dd, J= 9.3, 9.2 Hz, 1H), 6.76 (s, 1H), 6.37 (s, 1H), 4.99-4.85 (m, 1H), 4.10 (t, J= 8.6 Hz, 1H), 3.80-3.70 (m, 3H), 3.76 (t, J= 4.9 Hz, 4H), 3.12-3.02 (m, 1H), 2.98 (t, J= 4.7 Hz, 4H), 1.09 (d, J= 6.9 Hz, 6H), 1.06 (t, J= 6.7 Hz, 3H).13C NMR (150 MHz, DMSO-tL) 8 156.17, 155.83, 154.41, 152.34, 136.02, 133.87, 125.73, 119.69, 115.19, 114.58, 107.22, 102.69, 72.29, 66.60, 51.16, 49.03, 47.98, 31.12, 26.17, 23.0. HRMS calcd for C26H33FN3O6 [M + H]+502.2348, found 502.2353.

[0168] Reference Example 11 - Compound (29)

[0169] Reagents and conditions: (a) AC2O, PhMe, reflux, 16h (b) HNO3, H2SO4, 0 °C to r.t.,Ih (c) H2, Pd / C, DCM, r.t., 16h.

[0170] (S)-N-((3-(3-fluoro-4-morpholinophenyl)-2-oxooxazolidin-5-yl)methyl)acetamide (27)

[0171] Dissolved linezolid related amine 1 (2.5 g, 8.47 mmol, 1 eq) in toluene (37.5 mL), then acetic anhydride (2.02 mL, 21.18 mmol, 2.5 eq) was added. The resulting mixture was heated to reflux and react for 16h. After reaction finished, the white precipitation was collected through filtration to afford the desired product as white solid (2.3 g, 80 %). 'HNMR (300 MHz, DMSO-tL) 6 8.23 (t, J=5.9, IH), 7.47 (dd, J=15.0, 2.5, IH), 7.17 (dd, J=6.2, 2.7, IH), 7.06 (dd, J=9.6, 9.6), 4.77-4.62 (m, IH), 4.07 (t, J=9.0, IH), 3.78-3.63 (m, 5H), 2.99-2.90 (m, 4H), 1.83 (s, 3H).

[0172] (S)-N-((3-(5-fluoro-4-morpholino-2-nitrophenyl)-2-oxooxazolidin-5- yl)methyl)acetamide (28)

[0173] Suspended linezolid 27 (2 g, 5.93 mmol, leq) in sulfuric acid (20 mL) then fuming nitric acid (0.4 mL) was added dropwise under iced bath, then he resulting mixture was stirred for additional Ih. After reaction finished, the reaction was quenched by water, then adjust to pH>9. The mixture was extracted with EA / H2O three time and brine once. The organic layer was collected dried over MgSCh and removed solvent under reduced pressure to get the crude product. The crude product was used in next step without further purification.JH NMR (300 MHz, CDCI3) 8 7.63 (d, J=8.3, IH), 7.11 (d, J=12.3, IH), 6.25 (t, J=6.1, IH), 4.98-4.85 (m, IH), 4.07 (t, J=8.6, IH), 3.95-3.86 (m, 4H), 3.86-3.73 (m, 2H), 3.73-3.61 (m, IH), 3.24-3.14 (m, 4H), 2.11 (s, 3H).

[0174] (S)-N-( (3-(2-amino-5-fluoro-4-morpholinophenyl) -2-oxooxazolidin-5- yl)methyl)acetamide (29)

[0175] Dissolved crude product 28 in MeOH (20 mL), then 10 % Pd / C (cat.) was added. After three times degas, the resulting reaction mixture was stirred for 16 h under H2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was filtered through celite cake, washed with MeOH and collected the filtrate. The solvent was removed under reduced pressure to afford the crude product. The crude product was used in next step without further purification. 'H NMR (300 MHz, DMSO-tL) 6 8.26 (t, .7=5.9, IH), 6.93 (d, 7=13.3, IH), 6.39 (d, 7=8.7, IH), 5.00 (br s, 2H), 4.78-4.64 (m, IH), 4.20-2.09 (m, 2H), 3.85 (t, 7=8.6, IH), 3.78-3.67 (m, 4H), 3.56-3.46 (m, IH), 3.01-2.90 (m, 4H), 1.88 (s, 3H).

[0176] Compound (29) can be used as the initial reactant in the following examples.

[0177] Examples 12 and 13 - Compounds (31) and (34)

[0178] Reagents and conditions: (a) corresponding carboxylic acid, EDCi, HOBt, NMM, DMF, r.t., 16 h (b) H2, Pd / C, MeOH, r.t., 16 h (c) 1. 4-formyl cinnamic acid, MeOH, reflux, 16 h 2. NaBH4, 0 °C to r.t., 2 h (d) NH2OTHP, EDCi, HOBt, NMM, DMF, r.t., 16 h (e) 5 % TFA(MeOH), r.t., 16 h.

[0179] (S)-Nl-(2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)-N8-(benzyloxy)octanediamide (30)

[0180] Suspend 29 (200 mg, 0.57 mmol, 1 eq), 3-((benzyloxy)amino)-3-oxopropanoic acid (191 mg, 0.683 mmol, 1.2 eq), HOBt (92 mg, 0.683 mmol, 1.2 eq), EDCi (164 mg, 0.85 mmol, 1.5 eq) in dry DMF (5.7 mL) then NMM (0.1 ml, 0.85 mmol, 1.5 eq) was added. The reaction mixture was stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSO4and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH=30: 1) to afford the desired product as palewhite solid (80 mg, 23 %). 'H NMR (300 MHz, CD3OD) 5 7.64 (d, J=8.4, 1H), 7.50-7.32 (m, 5H), 7.38 (d, .7=12.7, 1H), 4.87 (s, 2H), 4.18 (t, 7=8.7, 1H), 3.91-3.79 (m, 5H), 3.66-3.58 (m,2H), 3.21-3.13 (m, 4H), 2.30 (t, J=7.4, 2H), 2.08 (t, 7=7.3, 2H), 2.05 (s, 3H), 1.68-1.51 (m, 4H), 1.42-1.26 (m, 4H).

[0181] (S)-Nl-(2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)-N8-hydroxyoctanediamide (31) (the titled compound)

[0182] Dissolved 30 (80 mg, 0.13 mmol, 1 eq) in MeOH (10 mL), then 10 % Pd / C (cat.) was added. After three times degas, the resulting reaction mixture was stirred for 16 h under H2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was filtered through celite cake, washed with MeOH and collected the filtrate. The solvent was removed under reduced pressure to afford the crude product. The crude product was purified by column chromatography (DCM:MeOH=10: l) to afford the desired product as pale-white solid (14 mg, 21 %); HPLC purity=97.07%; m.p =141.2 °C; *H NMR (300 MHz, CD3OD) 5 7.26 (d, 7=9.0, 1H), 7.19 (d, 7=12.9), 4.05 (t, 7=9.0, 1H), 3.88-3.84 (m, 4H), 3.77-3.67 (m, 1H), 3.60-3.57 (m, 2H), 2.43 (t, 7=7.5, 2H), 2.14 (t, 7=7.2, 2H), 1.75-1.44 (m, 4H), 1.44-1.41 (m, 4H).13C NMR (150 MHz, CD3OD) 5 172.65, 157.69, 147.69, 146.12, 144.15, 141.59, 140.36, 130.76, 126.93, 115.55, 114.94, 102.22, 73.21, 66.52, 50.67, 49.31, 48.42, 46.47, 41.75, 29.23, 22.78, 21.06. HRMS calcd for C24H35FN5O7 [M + H]+524.2515, found 524.2520.

[0183] (S,E)-3-(4-(((2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)amino)methyl)phenyl)acrylic acid (32)

[0184] Suspend 29 (200 mg, 0.57 mmol, 1 eq) and 4-formyl cinnamic acid (100 mg, 0.57 mmol, 1 eq) in MeOH (5.7 mL). The resulting reaction mixture was heated up to reflux and stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was cooled down to 0 °C, following with the addition of sodium caynoborohydride (240 mg, 2.27 mmol, 4 eq) portionwise. A pale-white solid precipitated immediately, the reaction mixture was stirred for additional 2 h. After reaction finished, collected the precipitate through filtration to afford the desired product as pale-white solid (300 mg, quant.).XH NMR (300 MHz, DMSO-tL) 8 8.29 (t, 7=5.6, 1H), 7.49 (d, 7=8.2, 2H), 7.37 (d,J=8.2, 2H), 7.23 (d, .7=15.8, 1H), 7.00 (d, 7=13.1, 1H), 6.41 (d, 7=15.8, 1H), 6.07 (d, 7=8.1, 1H), 5.00 (br s, 1H), 4.84-4.65 (m, 1H), 4.35 (d, 7=6.2, 2H), 3.90 (t, 7=8.6, 1H), 3.76-3.70 (m, 2H), 3.70-3.63 (m, 4H), 3.56-3.49 (m, 1H), 2.98-2.90 (m, 2H), 3.88-3.78 (m, 4H), 1.89 (s, 3H).

[0185] tetrahydro-2H-pyran-2-yl (E)-3-(4-(((2-((S)-5-(acetamidomethyl)-2- oxooxazolidin-3-yl)-4-fluoro-5-morpholinophenyl)amino)methyl)phenyl)acrylate (33)

[0186] Suspend compound 32 (300 mg, 0.57 mmol, 1 eq), HOBt (92 mg, 0.68 mmol, 1.2 eq), EDCi (164 mg, 0.85 mmol, 1.5 eq) in dry DMF (5.7 mL) then NMM (0.1 ml, 0.85mmol, 1.5 eq) was added. The reaction mixture was stirred for 10 min, then added <7-(tetrahydro-27 / - pyran-2-yl) hydroxylamine (80 mg, 0.68 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH=15: 1) to afford the desired product as pale-white solid (200 mg, 58 %).1HNMR (300 MHz, DMSO-tL) 88.29 (t, 7=6.0, 1H), 7.59-7.39 (m, 5H), 7.01 (d, 7=13.2, 1H), 7.48 (d, 7=16.0, 1H), 6.15-6.01 (m, 2H), 4.91 (br s, 1H), 4.84-4.69 (m, 1H), 4.38 (d, 7=6.0, 2H), 4.04-3.84 (m, 2H), 3.72-3.62 (m, 4H), 3.61-3.48 (m, 3H), 2.89-2.75 (m, 4H), 1.89 (s, 3H), 1.78-1.63 (m, 3H), 1.62-1.46 (m, 3H).

[0187] (S,E)-3-(4-(((2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)amino)methyl)phenyl)-N-hydroxyacrylamide (34) (the titled compound)

[0188] Dissolved 3 (200 mg, 0.33 mmol, 1 eq) in 5 % TFA in MeOH (10 mL) and stirred for 16 h at room temperature. The reaction is check by TLC. After the reaction finished, solvent was removed by reduced pressure. The residue was triturated with a mixture of diethyl ether and methanol, filtered, dried under vacuum to afford the desired product as pale-white solid (151 mg, 87 %); HPLC purity=99.67%; m.p =187.5 °C; 'H NMR (300 MHz, DMSO-tL) 6 8.29 (t, 7=6.3, 1H), 7.52 (d, 7=8.4, 2H), 7.46-7.39 (m, 3H), 7.00 (d, 7=13.2, 1H), 6.43 (d, 7=15.9),6.08-6.04 (m, 2H), 4.78-4.74 (m, 1H), 4.37 (d, J=6.3, 2H), 3.90 (t, J=8.7, 1H), 3.68-3.64 (m, 4H), 2.85-2.83 (m, 4H), 1.89 (s, 3H).13C NMR (150 MHz, DMSO ) 8 171.36, 164.12, 157.44, 147.66, 146.10, 143.15, 140.99, 139.46, 134.53, 128.87, 119.72, 116.96, 102.62, 73.66, 67.31, 51.58, 50.15, 47.17, 42.80, 23.70. HRMS calcd for C26H31FN5O6 [M + H]+528.2253, found 524.2272.

[0189] Examples 14 and 15 - Compounds (36) and (38)

[0190] Reagents and conditions: (a) 1. 4-formyl benzoic acid, MeOH, reflux, 16 h 2. NaBH4, 0 °C to r.t., 2 h (b) o-phenylenediamine, EDCi, HOBt, NMM, DMF, r.t., 16 h. (c) NH2OTHP, EDCi, HOBt, NMM, DMF, r.t., 16 h (d) 5 % TFA(MeoH), r.t., 16 h.

[0191] (S)-4-(((2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)amino)methyl)benzoic acid (35)

[0192] Suspend 29 (300 mg, 0.85 mmol, 1 eq) and 4-formylbenzoic acid (128 mg, 0.85 mmol, 1 eq) in MeOH (8.5 mL). The resulting reaction mixture was heated up to reflux and stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After starting material vanished, the reaction mixture was cooled down to 0 °C, added sodium cyanoborohydride (214 mg, 3.4 mmol, 4 eq), stirred for additional 2 h. After reaction finished,the solvent was removed under reduced pressure to afford crude product as yellow solid. The crude product was used in next step without further purification.JH NMR (300 MHz, CO3OD) 5 7.93 (d, J=7.5, 2H), 7.41 (d, J=8.2, 2H), 6.94 (d, J=12.8, 1H), 6.14 (d, J=8.3, 1H), 4.46 (s, 2H), 4.06 (t, .7=8.9, 1H), 3.79-3.70 (m, 4H), 3.67-3.53 (m, 3H), 2.92-2.85 (m, 4H), 2.03 (s, 3H).

[0193] (S)-4-(((2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)amino)methyl)-N-(2-aminophenyl)benzamide (36) (the titled compound)

[0194] Suspend crude product 35 (300mg, 0.62 mmol, leq), HOBt (100 mg, 0.74 mmol, 1.2 eq), EDCi (180 mg, 0.93 mmol, 1.5 eq) in dry DMF (6 mL) then NMM (0.1 ml, 0.93 mmol, 1.5 eq) was added. The resulting reaction mixture was stirred for 10 min, then added o- phenylenediamine (80 mg, 0.74mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSCU and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (EA) to afford the desired product as yellow solid (224 mg, 63 %); HPLC purity=96.79%; m.p.=144.5 °C; 'HNMR (300 MHz, DMSO-tL) 89.65 (s, 1H), 8.30 (t, .7=5.7, 1H), 7.94 (d, J=8.1, 2H), 7.52 (d, .7=8.4, 2H), 7.27-7.24 (m, 2H), 7.20-6.90 (m, 2H), 6.81 (d, J=8.1, 1H), 6.63 (t, .7=7.2, 1H), 6.15-6.07 (m, 2H), 4.79-4.76 (m, 1H), 4.44 (s, 2H), 3.92 (t, .7=8.7, 1H), 3.76-3.66 (m, 5H), 2.96- 2.93 (m, 1H), 2.86-2.83 (m, 4H), 1.90 (s, 3H).13C NMR (150 MHz, DMSO-tL) 6 170.50, 165.58, 156.62, 146.87, 145.31, 144.39, 143.54, 142.19, 140.21, 133.46, 128.24, 127.34, 123.80, 116.69, 115.97, 101.76, 72.86, 66.57, 50.91, 49.38, 49.03, 46.26, 42.00, 31.11, 22.90. HRMS calcd for C30H34FN6O5 [M + H]+577.2569, found 577.2582.

[0195] 4-(((2-((S)-5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)amino)methyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (37)

[0196] Suspend crude product 35 (400 mg, 0.82 mmol, l.Oeq), HOBt (132 mg, 0.98 mmol, 1.2 eq), EDCi (240 mg, 1.23 mmol, 1.5 eq) in dry DMF (8 mL) then NMM (0.14 ml, 1.23 mmol,1.5 eq) was added. The reaction mixture was stirred for 10 min, then added < -(tetrahydro-27 / - pyran-2-yl) hydroxylamine (115 mg, 0.98 mmol, 1.2 eq) to the reaction mixture stirred for 16 h under N2 atmosphere. The reaction was monitored by TLC. After reaction finished, the reaction mixture was extracted with EA / H2O three times and washed with saturated NH4Cl(aq) once, brine once. The organic layer was collected dried over MgSC and removed solvent under reduced pressure to get the crude product. The crude product was purified by column chromatography (DCM:MeOH=15: 1) to afford the desired product as pale white solid (220 mg, 46 %) 'HNMR (300 MHz, CD3OD) 57.76 (d, J=8.2, 2H), 7.50 (d, J=8.2, 2H), 6.96 (d, J=12.8, 1H), 6.10 (d, J=8.2), 5.07 (s, 1H), 4.46 (s, 2H), 4.21-3.99 (m, 2H), 3.80-3.70 (m, 4H), 3.70-3.55 (m, 4H), 2.93-2.82 (m, 4H), 2.03 (s, 3H), 1.98-1.76 (m, 3H), 1.77-1.52 (m ,3H).

[0197] (S)-4-(((2-(5-(acetamidomethyl)-2-oxooxazolidin-3-yl)-4-fluoro-5- morpholinophenyl)amino)methyl)-N-hydroxybenzamide (38) (the titled compound)

[0198] Dissolved 37 (220 mg, 0.38 mmol, 1 eq) in 5 % TFA in MeOH (10 mL) and stirred for 16 h at room temperature. The reaction is check by TLC. After the reaction finished, solvent was removed by reduced pressure. The residue was triturated with a mixture of ethyl acetate and methanol, filtered, dried under vacuum to afford the desired product as orange solid (176 mg, 93 %); HPLC purity=97.19%; m.p.=190.2 °C; 'H NMR (300 MHz, CD3OD) 5 7.73 (d, J=8.1, 2H),7.51 (d, J=7.8, 2H), 6.96 (d, J=12.9, 1H), 6.11 (d, J=8.4, 1H), 4.50 (s, 2H), 4.05 (t, J=8.7, 1H), 3.78-3.74 (m, 4H), 3.70-3.59 (m, 3H), 2.91-2.87 (m, 4H), 2.03 (s, 3H).13C NMR (150 MHz, CD3OD) 5 173.52, 172.60, 171.52, 156.51, 153.41, 151.78, 139.60, 130.27, 124.94, 120.03, 116.07, 113.80, 73,24, 66.48, 50.49, 49.43, 41.85, 35.87, 32.25, 28.47, 25.21, 21.08. HRMS calcd for C24H29FN5O6 [M + H]+502.2096, found 524.2103.

[0199] Example 16: In vitro cytotoxicity studies - cytotoxicity of the compounds as disclosed against TMZ-Resistant GBM.

[0200] To confirm the cytotoxicity of the functional domains of the compounds against glioblastoma (GBM), the compounds as synthesized were administered to TMZ-resistant GBMcell lines A172R and U87MGR at concentrations ranging from 0 to 80 pM over a 72-hour period. The cytotoxic effects of these compounds were assessed using the MTT assay (Figure 2A)

[0201] The results indicated that Compound (4) (MPT1A089), Compound (12) (MPT1A092) and Compound (24) (MPT1A097) exhibited the capacity to induce cytotoxicity in TMZ-resistant GBM.

[0202] Given the remarkable cytotoxicity of Compound (4) (MPT1 A089), it was taken as the representative model compound. Nevertheless, other potential compounds may exhibit similar efficacy. To validate that the cytotoxic effect of Compound (4) (MPT1 A089) on GBM is not attributed to linezolid alone, the applicant separately treated TMZ-resistant GBM cell line PT#3-R with Compound (4) (MPT1 A089) and linezolid. The results revealed that linezolid, at concentrations ranging from 80-100 pM, did not induce cytotoxicity in resistant GBM cells. In contrast, Compound (4) (MPT1A089), at concentrations of 20 pM and above, exhibited a significant cytotoxic effect on resistant GBM cells, with cytotoxicity increasing proportionally with Compound (4) (MPT1A089) concentration. This confirms that the cytotoxicity of Compound (4) (MPT1A089) against GBM is primarily attributable to its specific structure (Figure 2B), not the linezolid moiety. Furthermore, the applicant compared the cytotoxic effects of Compound (4) (MPT1 A089) on various wild-type GBM cell lines, including U87MG, PT#3, and LN229, with TMZ-resistant GBM cell lines, including U87MG-R, PT#3-R, and T98G (MGMT-positive). Compound (4) (MPT1 A089) exhibited outstanding cytotoxicity, and the IC50 values remained consistent regardless of TMZ resistance (Figure 2C). Additionally, the applicant further confirmed the efficacy of Compound (4) (MPT1 A089) in GBM treatment. While the induced IC50 in primary cultures was 143.1, it displayed inhibitory effects on proliferation and induced cell death in GBM, TMZ-resistant GBM, and patient-derived TMZ- resistant GBM cells, with IC50 values ranging from 10.32 to 30.58. Notably, the IC50 valuefor primary mouse astrocytes was higher, indicating a greater selectivity of Compound (4) (MPT1A089) against GBM cells (Figure 2D).

[0203] Example 17 - Inhibitory effect on Class I HDACs

[0204] To assess the inhibitory potential of Compound (4) (MPT1A089) on Histone Deacetylases (HDACs), enzyme inhibition assays were conducted and they unequivocally revealed effective inhibition of Class I HDACs by Compound (4) (MPT1A089), specifically targeting HD AC 1, 2, and 8 (see, Table 1 below). Furthermore, a comprehensive analysis of The Cancer Genome Atlas Glioblastoma Multiforme (TCGA-GBM) and Chinese Glioma Genome Atlas (CGGA-GBM) databases unveiled notably elevated mRNA expression levels of Class I HDACs within GBM compared to normal brain tissue. Intriguingly, as primary GBM progresses to the recurrent stage, the abundance of Class I HDACs further escalates. Moreover, the abnormally heightened expression of Class I HDACs strongly correlates with unfavorable survival outcomes in GBM patients (Figure 3A). These collective findings underscore the pivotal role of Class I HDACs in fueling GBM growth and fostering resistance to temozolomide (TMZ). To validate inhibitory effects of Compound (4) (MPT1A089) on Class I HDACs and the resulting acetylation changes in GBM cells, we conducted an analysis of Histone H3 and H4 acetylation levels after treatment with Compound (4) (MPT1 A089). The results unveiled substantial acetylation modifications at distinct sites on Histone H3 and H4 following Compound (4) (MPT1 A089) treatment (Figure 3B-C).aThese assays were conducted by the Reaction Biology Corporation, Malvern, Pennsylvania. All compounds were dissolved in DMSO and tested in a 10-dose IC50 mode with 3-fold serial dilution starting at 10 pM.bN / A indicates that the tested compound could not fit into IC50 curve.Table 1

[0205] Example 18 - Inhibition of GBM cell growth (via DNA function inhibition)

[0206] To investigate the mechanism of action of Compound (4) (MPT1A089), the applicant conducted a RNA-seq on TMZ-resistant GBM cells treated with the compound. In comparison to treatment with linezolid, a 72-hour exposure to Compound (4) (MPT1A089) resulted in the upregulation of 1105 genes and the downregulation of 1191 differential expression genes (DEGs), indicating a significant impact on cellular proliferation dynamics (Figure 4A). Further exploration through gene set enrichment analysis (GSEA) unveiled a profound suppression of DNA replication and mitotic nuclear division capabilities, accompanied by a substantial reduction in the expression of genes associated with these processes (Figures 4B, 4C, 4E). To corroborate these findings, cell cycle experiments were conducted, confirming a decrease in the number of cells in the G0 / G1 phase upon Compound (4) (MPT1 A089) treatment, while there was an increase in cells in the G2 / M phase and the sub- G1 phase (Figure 4D). These results collectively suggest that Compound (4) (MPT1A089) treatment disrupts normal DNA physiological functions, induces cellular damage, and consequently inhibits cell growth.

[0207] Example 19 - Disruption of DNA repair mechanisms in GBM cells by suppressing DNA-repaired proteins.

[0208] To comprehensively evaluate the impact of Compound (4) (MPT1A089) treatment on DNA damage, the applicant conducted a DNA Homologous Recombination (HR) assay. Homologous recombination repair plays a crucial role in repairing DNA double-strand breaks (DSBs). However, upon exposure to Compound (4) (MPT1A089), the applicant observed a substantial decrease in DNA HR repair capability with increasing drug concentrations, indicating the inhibition of DNA repair post-compound treatment (Figure 5A-B). Further mechanistic analysis unveiled a significant increase in the phosphorylation of ataxia telangiectasia mutated kinase (ATM), a pivotal protein in the DSB repair mechanism. This suggests an activation of the physiological response to DSBs induced by Compound (4) (MPT1A089). Intriguingly, downstream proteins crucial for the repair process, including carboxy-terminal binding protein (CtIP), Replication protein A 32 kDa subunit (RPA32), and DNA repair protein RAD51 homolog 1 (RAD51), exhibited significant suppression, signifying that Compound (4) (MPT1A089) inhibits the downstream mechanisms of DNA repair (Figure 5C). Furthermore, when treated with Cycloheximide (CHX), a protein synthesis inhibitor disrupting the translocation step in the protein synthesis process, the applicant observed a less pronounced reduction in Rad51 and CtIP protein levels. In contrast, under Compound (4) (MPT1A089) treatment, Rad51 and CtIP protein expression rapidly and significantly decreased over time (Figure 5D-E). This result suggests that Compound (4) (MPT1 A089) induces the degradation of Rad51 and CtIP, ultimately leading to cellular death due to the compromised stability of DNA repair proteins.

[0209] Example 20 - Induction of ubiquitination and degradation of Rad51 proteins in GBM Cells.

[0210] To Ubiquitination, a pivotal physiological process responsible for tagging proteins for degradation, plays a crucial role in cellular regulation. Upon treatment with Compound (4) (MPT1A089), the applicant conducted immunoprecipitation using an anti-Rad51 antibody to investigate the ubiquitination status of the Rad51 protein. The results revealed a significantincrease in Rad51 ubiquitination induced by Compound (4) (MPT1 A089), subsequently leading to protein degradation (Figure 6A). To ascertain that the phenomenon was not an off-target effect, the applicant further dissected Rad51 into three functional fragments based on its sequence (Figure 6H). The applicant performed immunoprecipitation using a GFP protein to detect ubiquitinated proteins. The findings clearly demonstrated that overexpression of Rad51 fragment 3 in pEGFP-RAD51-T3 exhibited the most prominent ubiquitination when exposed to Compound (4) (MPT1A089) (Figure 6B-D). Moreover, through introducing mutations at ubiquitination sites within fragment 3, specifically changing lysine residues to alanine (Figure 6H), the applicant observed a reduction in protein ubiquitination at both mutation 1 and mutation 2 positions (Figure 6E-G). These compelling results substantiate that MPT1A089 indeed triggers ubiquitination of DNA repair proteins, leading to their subsequent degradation. Consequently, this loss of double-strand break repair capabilities culminates in GBM cell death.

[0211] Example 21 - Efficiency in treating in vivo GBM and assessing blood-brain barrier penetration.

[0212] The development of drugs for GBM has become increasingly challenging due to the selective control of substances entering the brain imposed by the blood-brain barrier (BBB). To further validate the effectiveness of Compound (4) (MPT1 A089) in in vivo GBM treatment, the applicant employed UPLC analysis. Fortunately, no significant overlap occurred comparing Compound (4) (MPT1A089) signal with either wild-type mice plasma or brain tissue extraction, which indicated the specificity of the established analysis method. Subsequently, the applicant conducted intraperitoneal injections of TMZ at a dose of 60 mg / kg and Compound (4) (MPT1 A089) at a concentration of 360 mg / kg. After a 30-minute interval, the applicant performed UPLC analysis on plasma and brain tissue extract, which revealed the presence of Compound (4) (MPT1A089) signals in both plasma and brain tissue extract, in contrast to the sham and solvent control mice (Figure 7A). The corresponding Compound (4)(MPT1A089) signal observed on LC spectrum was then further analyzed with mass detection at m / z=471.0 (M + H+) (Figure 7B).

[0213] The resulting retention time showed correspondence to the LC result, which, distinctly identifying the peak of Compound (4) (MPT1A089) in brain tissue extract as compared to the DMSO-injected control, thereby demonstrating the potential thereof to traverse the BBB and enter the brain as a potential therapeutic agent. To quantify the penetration ability of Compound (4) (MPT1A089) through the BBB and ensure precise analysis during sampling, the establishing bioanalysis method has been validated according to the request by US Food and Drug Administration (FDA) bioanalytical method validation, guidance for industry (FDA, 2018). The assessed validation, including specificity, linearity, lower limit of detection (LLOD), lower limit of quantification (LLOQ), precision, accuracy, and matrix effect has all reached the criteria (data not shown). The detected concentrations of Compound (4) (MPT1A089) in plasma and brain tissue were 891 ng / mL and 656 ng / mL, respectively, with a Brain / Plasma ratio of 1.27 (Figure 7C). In orthotopic GBM experiments using TMZ-resistant U87MG cells in mice, Compound (4) (MPT1A089) demonstrated potential to extend the survival rate in the in-situ transplantation model of GBM cells (Figure 7D). Post-mortem brain tissue sections further revealed the effective inhibition of TMZ-resistant GBM cell growth in the brain by MPT1A089 (Figure 7E). These compelling results provide substantial evidence that Compound (4) (MPT1A089) has the capability to penetrate the blood-brain barrier and holds promise as a candidate for treating TMZ-resistant GBM.

[0214] In conclusion, numerous compounds were prepared based on linezolid-structure for repurposing into the realm of GBM. The synthesized compounds were tested against either primary or resistance GBM cell lines. Among the same, Compound (4) (MPT1A089) displayed the best cell growth inhibition. The result indicated potential for further structural optimization. For the investigation of mechanism of action, Compound (4) (MPT1 A089) was further conducted with a comprehensive biological evaluation. Indeed, Compound (4)(MPT1A089) displayed HDAC enzyme inhibition and induced cell cycle arrest, which is similar to the reported HDAC inhibitors. Interestingly, RNA-seq result suggested that Compound (4) (MPT1 A089) disrupted the DNA repair, which was then investigated in detail. Through HR assay the ability of DNA repair disruption was validated and found to be caused by the absence of RAD51 and CtIP protein and lead to the ubiquitination of DNA repair proteins. Moreover, the orthotopic model displayed an effective therapeutic effect and satisfying BBB penetration, which was again validated through normal mouse models. Overall, Compound (4) (MPT1A089) with promising results in both in vitro and in vivo studies, proved highly potent to become a novel therapeutic strategy for GBM.

[0215] A person of ordinary skill in the art of the subject disclosure should understand that variations and modification may be made to the teaching and the disclosure of the subject disclosure without departing from the spirit and scope of the subject application. Based on the disclosure, the subject application intends to cover any variations and modification thereof with the proviso that the variations or modifications fall within the scope as defined in the appended claims or equivalents thereto.

Claims

We claim:

1. A compound of Formula (I),or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: p is 0 or 1, with the proviso that when p is 0, A has the definition of Ai, and when p is 1, A is acetyl;Ai has the formulathe formula of *-L-Rb, wherein * denotes the linking position to N;-L- is selected from the group consisting of-CH2-, -C(=O)- and -S(=O)2-;-Sp- is selected from the group consisting of Ci-ealkylene,and , wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety - C(=O)NHRa;Ra is -OH or 2-aminophenyl;Rb is selected from the group consisting ofand, wherein ** denotes the linking position to L; andRi is H, methyl or ethyl.

2. The compound of claim 1, wherein Formula (I) is Formula (IA):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein the definition of Ai and Ri are as defined in Claim 1.

3. The compound of claim 1, wherein Formula (I) is Formula (IB):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein the definition of Ai is as defined in Claim 1.

4. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, whereindenotes the linking position to N, -L- is -CH2-, -Sp- is, wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa.

5. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, whereinwherein * denotes the linking position to N, -L- is -C(=O)-, -Sp- is Ci-ealkylene.

6. The compound of claim 5, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein -Sp- is 1,6-w-hexanylene.

7. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, whereindenotes the linking position to N, -L- is -S(=O)2-, -Sp- is wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa.

8. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein Ai is *-L-Rb, wherein * denotes the linking position to N, and L is -C(=O)-.

9. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, whereinwherein *denotes the linking position to N, -L- is -CH2-, -Sp- is wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa, Ra is -OH and Ri is H.

10. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, whereinwherein *denotes the linking position to N, -L- is -CH2-, -Sp- is , wherein** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=0)NHRa, Rais -OH or 2-aminophenyl, preferably 2-aminophenyl, and Ri is H.

11. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt, hydrate,solvate, prodrug or isomer thereof, wherein Ai is, wherein * denotes the linking position to N, -L- is -C(=O)-, -Sp- is 1,6-w-hexanylene, Rais -OH and Ri is H.

12. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, whereinwherein *denotes the linking position to N, -L- is -S(=O)2-, -Sp- is wherein ** denotes the linking position to L and *** denotes the linking position to the carbonyl in the moiety -C(=O)NHRa, Ra is -OH and Ri is H.

13. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein Ai is *-L-Rb, wherein * denotes the linking position,-L- is C(=O) and Rb iswherein ** denotes the linking position toL, and Ri is H.

14. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein Ai is *-L-Rb, wherein * denotes the linking position,-L- is -C(=O) and Rb is, wherein ** denotes the linking position to L, and Ri is H.

15. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, hydrate, solvate,prodrug or isomer thereof, wherein Ai is *-L-Rb, wherein * denotes the linking position,-L- is -C(=0) and Rb isto L, and Ri is H, methyl or ethyl.

16. The compound of claim 1, selected from the group consisting of:Compound (22)or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof.

17. A pharmaceutical composition comprising a compound of any one of claims 1-16, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, and a pharmaceutically acceptable excipient.

18. A method of treating a cancer or a recurrent or metastatic cancer in a patient in need, comprising administering an effective amount of a compound of Formula (I), Formula (IA) or Formula (IB), or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of anyof foregoing to said subject.

19. The method according to Claim 18, wherein the cancer is a brain cancer or a recurrent or metastatic brain cancer.

20. The method according to Claim 18 or 19, wherein the cancer is recurrent or metastatic glioblastoma (GBM).

21. The method of any one of Claims 18-20, wherein the compounds of Formula (I), Formula (IA) or Formula (IB) exhibit inhibitory effects on HD AC, preferably Class 1 HD AC.

22. The method of any one of Claims 18-20, wherein the the compounds of Formula (I), Formula (IA) or Formula (IB) exhibit inhibitory effects on heat shock protein 90 (Hsp90) inhibitors.

23. The method of any one of Claims 18-20, wherein the compounds of Formula (I), Formula (IA) or Formula (IB) act as antitubulin agents.

24. The method of any one of Claims 18-20, wherein the compounds of Formula (I), Formula (IA) or Formula (IB) act as anti-cyclooxygenase (COX) agents, preferably anti-COX-2 agents.

25. The method of any one of Claims 18-20, wherein the the compounds of Formula (I), Formula (IA) or Formula (IB) can cross the blood-brain barrier (BBB).

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