HK40043214B - Compound acting as antibiotics - Google Patents

Description

Compounds as antibiotics

Technical Field

This invention belongs to the field of pharmaceutical technology and relates to compounds as antibiotics, their pharmaceutically acceptable salts, esters, their prodrugs, their solvates, deuterated derivatives or stereoisomers thereof, methods for preparing these compounds, and pharmaceutical compositions containing these compounds; this invention also relates to the use of said compounds for the treatment and/or prevention of infectious diseases, especially for the treatment and/or prevention of infectious diseases caused by Gram-negative bacteria.

Background Technology

In recent years, the widespread use of highly effective broad-spectrum antibacterial drugs has created selective pressure on bacteria, leading to a continuous increase in drug-resistant and drug-resistant strains of common Gram-negative bacilli. Clinically, Enterobacteriaceae bacteria that produce extended-spectrum β-lactamases (ESBLs) have even emerged, as well as multidrug-resistant Pseudomonas aeruginosa and Acinetobacter aeruginosa that have shown resistance to all β-lactam and quinolone antibacterial drugs.

Lipid A is the membrane-bound region of lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria (such as Pseudomonas aeruginosa and Acinetobacter baumannii). It is a crucial component protecting bacteria against external factors (such as antibiotics). Furthermore, it is a potent endotoxin that can cross the intestinal mucosal barrier into the bloodstream, causing fatal septic shock and is a significant contributing factor to Gram-negative bacterial infections. Therefore, it is generally believed that inhibiting lipid A biosynthesis can control diseases caused by Gram-negative bacteria. UDP-3-O-(R-hydroxytetradecanoyl)-N-acetylglucosamine deacetylase (LpxC) is a zinc metalloenzyme that catalyzes the second step of lipid A biosynthesis in Gram-negative bacteria. It is an essential enzyme for the survival of Gram-negative bacteria; therefore, inhibiting LpxC can suppress bacterial growth and thus exert a therapeutic effect on diseases caused by Gram-negative bacteria.

Patents WO2004062601 A2 and others disclose a series of LpxC inhibitors such as CHIR-090, and WO2008154642A2 also discloses some LpxC inhibitors such as ACHN-975. Patents WO2011132712 A1 and others also disclose a series of LpxC inhibitors, some of which have the following structures:

These existing compounds exhibit some antibacterial activity against bacteria, particularly Gram-negative bacteria; however, there is still room for improvement in their antibacterial activity.

Summary of the Invention

Through in-depth and diligent research, the inventors have discovered a novel antibiotic compound, its pharmaceutically acceptable salt, ester, prodrug, solvate, deuterated derivative, or stereoisomer thereof (hereinafter also referred to as the compound of this invention), which is an inhibitor of UDP-3-O-(R-hydroxytetradecanoyl)-N-acetylglucosamine deacetylase (LpxC). The compound of this invention exhibits excellent antibacterial activity, particularly against Gram-negative bacteria.

Specifically, the present invention provides compounds represented by the following general formula (I), pharmaceutically acceptable salts thereof, esters thereof, prodrugs thereof, solvates thereof, deuterated derivatives thereof, or stereoisomers thereof:

in,

W indicates

R ¹ is selected from -(CH ₂ ) _0-4 C(R ^1a , R ^1b )(CH ₂ ) _0-4 OR ³ , -(CH ₂ ) _0-4 C(O)NR ⁴ R ⁵ , -(CH ₂ ) _0-4 C(R ^1a , R ^1b )NR ⁴ R ⁵ , -(CH ₂ ) _0-4 C(R ^1a , R ^1b )(CH ₂ ) _0-4 S(O) _0-2 R ⁶ and -(CH ₂ ) _0-4 C(R ^1a , R ^1b )(CH ₂ ) _0-4 SC(O)R ⁷ ,

Among them, ^R1a , ^R1b , ^R3 , ^R4 , ^R5 , ^R6 and ^R7 are each independently selected from H, _C1-6 alkyl and OH;

R ^1′ is selected from hydrogen and _C1-6 alkyl groups;

_A1 , _A2 , and _A3 are each independently selected from CH and heteroatoms;

X and Y are each independently selected from benzene ring groups, 3-8 membered unsaturated heterocyclic groups, alkenyl groups, alkynyl groups, and...

^R2 is

B is absent or represents -( _C1-8 alkyl)-, wherein the 0-2 carbon atoms of the -( _C1-8 alkyl)- are optionally replaced by O or NR ⁸ .

C is selected from _C1-6 alkyl, ^-OR9 , ^{-NR9R9 '} , phenyl, and 3-8 membered saturated and/or unsaturated heterocyclic groups.

^R8 , ^R9 , and R9 ^' are each independently selected from hydrogen atoms, _C1-6 alkyl groups, and _C3-6 cycloalkyl groups;

m can be 0, 1, 2, or 3.

In another embodiment of the invention, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof, a prodrug thereof, a solvate thereof, a deuterated thereof, or a stereoisomer thereof is provided, wherein,

W indicates

^R1 is selected from -( _CH2 ) _O- 4C( ^R1a , ^R1b )( _CH2 )O ^-4OR3 _, - ⁽ _CH2 ) _O- ^4C (O) ^NR4R5 and -( _CH2 )O- _4C ( ^R1a , ^R1b ) ^NR4R5 , wherein ^{R1a, R1b} , ^R3 , ^R4 and ^R5 are each independently selected from H, _C1-4 alkyl and OH;

R ^1′ is selected from hydrogen and _C1-6 alkyl groups;

_A1 , _A2 , and _A3 are each independently selected from CH and heteroatoms;

X and Y are each independently selected from benzene ring groups, 5-8 membered unsaturated heterocyclic groups, alkenyl groups, alkynyl groups, and...

^R2 is

B is absent or represents -( _C1-8 alkyl)-, wherein the 0-2 carbon atoms of the -( _C1-8 alkyl)- are optionally replaced by O or NR ⁸ .

C is selected from _C1-6 alkyl, ^-OR9 , ^{-NR9R9 '} , phenyl, and 3-8 membered saturated and/or unsaturated heterocyclic groups.

^R8 , ^R9 , and R9 ^' are each independently selected from hydrogen atoms, _C1-6 alkyl groups, and _C3-6 cycloalkyl groups;

m can be 0, 1, or 2.

In another embodiment of the invention, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof, a prodrug thereof, a solvate thereof, a deuterated thereof, or a stereoisomer thereof is provided, wherein,

W indicates

^R1 is selected from -C ^{( R1a} , ^R1b ) ^OR3 and -C(O) ^NR4R5 , wherein ^R1a , ^R1b , ^R3 , ^R4 and ^R5 are each independently selected from hydrogen atoms, methyl and hydroxyl groups;

R1 ^′ is selected from hydrogen and _C1-4 alkyl;

_A1 , _A2 , and _A3 are each independently selected from CH and N;

X and Y are each independently selected from benzene ring groups, 5-6 membered unsaturated heterocyclic groups, alkenyl groups, alkynyl groups, and...

^R2 is

B either does not exist or represents -( _C1-4 alkyl)-.

C is selected from -OR ⁹ and -NR ⁹ R ^9' .

^R9 and R9 ^' are each independently selected from hydrogen atoms, methyl, ethyl, and isopropyl groups;

m can be 0, 1, or 2.

In another embodiment of the invention, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof, a prodrug thereof, a solvate thereof, a deuterated thereof, or a stereoisomer thereof is provided, wherein,

W indicates

^R1 is selected from -C ^{( R1a} , ^R1b ) ^OR3 and -C(O) ^NR4R5 , wherein ^R1a , ^R1b , ^R3 , ^R4 and ^R5 are each independently selected from hydrogen atoms, methyl and hydroxyl groups;

R ^1' is selected from hydrogen and _C1-4 alkyl;

_A1 , _A2 , and _A3 are each independently selected from CH and N;

X and Y are each independently selected from the following groups: benzene ring group, pyrrole ring group, imidazole ring group, pyrazole ring group, 1,2,3-triazole ring group, 1,2,4-triazole ring group, tetrazolium ring group, thiophene ring group, thiazole ring group, isothiazole ring group, 1,2,4-thiadiazole ring group, furan ring group, oxazole ring group, isoxazole ring group, 1,2,4-oxadiazole ring group, pyridine ring group, alkenyl group, alkynyl group.

^R2 is ( _CH2 ) _1-4OH ;

m can be 0, 1, or 2.

In another embodiment of the invention, a compound of general formula (I), a pharmaceutically acceptable salt thereof, an ester thereof, a prodrug thereof, a solvate thereof, a deuterated thereof, or a stereoisomer thereof is provided, wherein,

W indicates

^R1 is selected from -C(H, _CH3 )OH and -C(O) _NHCH3 ;

R ^1' is selected from hydrogen, methyl, and ethyl;

_A1 , _A2 , and _A3 are each independently selected from CH and N;

X and Y are each independently selected from benzene ring groups, pyrrole ring groups, imidazole ring groups, pyrazole ring groups, 1,2,3-triazole ring groups, 1,2,4-triazole ring groups, tetrazolium ring groups, furan ring groups, oxazole ring groups, isoxazole ring groups, 1,2,4-oxadiazole ring groups, pyridine ring groups, alkenyl groups, alkynyl groups, etc.

^R2 is _-CH2OH ;

m is 0 or 1.

In another embodiment of the invention, compounds of general formula (II), pharmaceutically acceptable salts thereof, esters thereof, prodrugs thereof, solvates thereof, deuterated derivatives thereof, or stereoisomers thereof are provided:

Wherein, W represents -( _C0-8 alkyl group optionally substituted with a substituent)-C(O)-N(R1 ^′ )-( _C1-8 alkyl group optionally substituted with ^R1 )-C(O)-N(H)-OH;

^R1 is independently selected from _C1-8 alkyl, -( _{C0-8 alkyl)C(O)NR4R5, -(C0-8} ^alkyl )S(O) _1-2R3 and -( _C0-8 alkyl)S ^{(O)1-2NR4R5 , wherein R3 , R4} and ^R5 are independently selected from hydrogen atom, halogen atom, _{C1-8 alkyl} , _{C2-8 alkenyl} , and _C2-8 alkynyl;

R ^1′ is selected from hydrogen atom, hydroxyl group, halogen atom, carboxyl group, optional substituent-substituted _C1-8 alkyl group, optional substituent-substituted _C2-8 alkenyl group and optional substituent-substituted _C2-8 ynyl group, or R ^1′ is connected to a carbon atom on an optional substituent-substituted _C0-8 alkyl group in the W group and together with the N, C(O) group to form a 5-6 membered heterocyclic group;

_A1 , _A2 , _A3 , _A4 , _A5 and _A6 are each independently selected ^{from CRa} , ^NRC , O and S;

X and Y are each independently selected from a bond, a 5-14 membered heteroaryl group substituted by a substituent, a 6-14 membered aromatic group substituted by a substituent, -(C＝C)-, -(C≡C)-, =N-, -C(O)-NR ^c- , wherein at least one of X and Y is not a bond;

_P1 , _P2 , _P3 , _P4 and _P5 are each independently selected ^{from CRaRa} , ^NRC , O and S;

m represents an integer between 0 and 4;

Each occurrence of R ² is represented independently.

Each occurrence of B independently represents a bond, or is independently selected from C1-8 alkyl groups in which at least one carbon atom is substituted by at least one group of S, O and NR ^c and/or optionally substituted by a substituent, _C2-8 alkenyl groups in which at least one carbon atom is substituted by at least one group of S, O and NR ^c and/or optionally substituted by a substituent, and _C2-8 alkynyl groups in which at least one carbon atom is substituted by at least one group of S, O _and NR ^c and/or optionally substituted by a substituent;

Each C independently represents a hydrogen atom, cyano, mercapto, halogen atom, carboxyl, nitro, optional substituted _C1-8 alkyl, optional substituted _C1-8 alkoxy, -OR ^c , -NR ^c R ^c , optional substituted 3-12 membered cycloalkyl, optional substituted 3-12 membered heterocyclic, optional substituted 5-14 membered heteroaryl, and optional substituted 6-14 membered aryl.

^Ra may or may not exist, or each of them may be independently selected from hydrogen, cyano, mercapto, halogen, carboxyl, nitro, -OR ^c , -NR ^c R ^c , -N(OH)R ^c , -C(O)R ^d , -C(O)OR ^c , -C(O)NR ^c R ^c , -OC(O)NR ^c R ^c , -NR ^c C(O)OR ^c , -NR ^c C(O)R ^d , -S(O) _1-2 -NR ^c R ^c , -S(O) _1-2 R ^d , -NR ^c S(O) _1-2 R ^d , -S(O) _1-2 -OR ^c , _C1-8 alkyl groups optionally substituted with substituents, -( _C1-8 alkyl groups optionally substituted with substituents)OR ^c , -(C1-8 alkyl groups optionally substituted with substituents)NR ^c R ^c , _C1-8 alkyl groups optionally substituted with substituents _1-8 alkoxy, optional substituent-substituted _C2-8 alkenyl, optional substituent-substituted _C2-8 alkynyl, optional substituent-substituted 3-12 cycloalkyl, optional substituent-substituted 3-12 heterocyclic, optional substituent-substituted 6-14 aryl and optional substituent-substituted 5-14 heteroaryl.

R c  may be absent individually, or each Rc may be independently selected from hydrogen atom, halogen atom, carboxyl group, -C(O)R d , -C(O)OR  b  , -C(O)NR  b R b , -S(O) 1-2 -NR bRb , -S(O) 1-2 -Rd, -S(O) 1-2 -OR b , optional substituted C 1-8  alkyl, -(optionally substituted C 1-8 alkyl)ORb, -(optionally substituted C 1-8 alkyl)NR b  R b, optional substituted C 2-8  alkenyl, optional substituted C2-8 alkynyl, optional substituted 3-12-membered cycloalkyl, optional substituted 3-12-membered heterocyclic, optional substituted 6-14-membered aryl, and optional substituted 5-14-membered heteroaryl; or two Rc When c is attached to the same atom, the two R c together with the attached atom form a 3-12 membered heterocycle that can be optionally substituted by a substituent.

Each ^Rb , when appearing, is independently selected from a hydrogen atom, a halogen atom, a carboxyl group, a sulfonic acid group, a _C1-8 alkyl group optionally substituted with a substituent, a _C1-8 alkylsulfonyl group optionally substituted with a substituent, a _C1-8 alkylsulfinyl group optionally substituted with a substituent, a 3-12 membered cycloalkyl group optionally substituted with a substituent, a 3-12 membered heterocyclic group optionally substituted with a substituent, a 6-14 membered aryl group optionally substituted with a substituent, and a 5-14 membered heteroaryl group optionally substituted with a substituent; or, when two ^Rb are attached to the same atom, the two ^Rb together with the attached atom form an optionally substituted 3-12 membered heterocycle.

Each time R ^d appears, it is independently selected from hydrogen atom, hydroxyl group, mercapto group, halogen atom, carboxyl group, nitro group, amino group, optional substituent-substituted _C1-8 alkylamino group, (optionally substituent-substituted _C1-8 alkyl) _2amino group, sulfonic acid group, optional substituent-substituted _C1-8 alkyl group, optional substituent-substituted _C1-8 alkoxy group, optional substituent-substituted _C1-8 alkylsulfonyl group, optional substituent-substituted _C1-8 alkylsulfinyl group, optional substituent-substituted 3-12 membered cycloalkyl group, optional substituent-substituted 3-12 membered heterocyclic group, optional substituent-substituted 6-14 membered aryl group, and optional substituent-substituted 5-14 membered heteroaryl group.

The substituents in "optionally substituted" are each independently selected from: hydroxyl, mercapto, carboxyl, cyano, nitro, amino, halogen atom, sulfonic acid group, _C1-8 alkyl, _C1-8 alkoxy- _C1-8 alkyl, hydroxy- _C1-8 alkyl, amino- _C1-8 alkyl, carboxyl- _C1-8 alkyl, ester- _C1-8 alkyl, _C1-8 alkoxy, hydroxy- _C1-8 alkoxy, amino- _C1-8 alkoxy, carboxyl- _C1-8 alkoxy, _C1-8 alkyl- _C1-8 alkoxy, _C1-8 alkoxy- _C1-8 alkoxy, ester- _C1-8 alkoxy, _C1-8 alkylamino, ( _C1-8 alkyl) _2amino , amino- _C1-8 alkylamino, (amino- _C1-8 alkyl) _2amino , _C1-8 alkyl ester, aminocarbonyl, _C1-8 alkylaminocarbonyl, ( _C1-8 alkyl) _2- aminocarbonyl, _C1-8 alkylcarbonyl, C1-8 alkylcarbonyloxy, _C1-8 alkylcarbonylamino, _C1-8 alkylsulfonylamino, halo _-C1-8 alkyl, halo _-C1-8 alkoxy, _C1-8 alkylsulfonyl, _C1-8 alkylsulfinyl, _C1-8 alkylthio, _3-12 membered cycloalkyl, 6-14 membered aryl, 3-12 membered heterocyclic, 5-14 membered heteroaryl and oxo,

The double bonds in the five-membered ring (the ring marked with P in general formula (II), and hereinafter referred to by the same symbol) and the six-membered ring (the ring marked with A in general formula (II), and hereinafter referred to by the same symbol) are double bonds that are arbitrarily present in the ring;

The condition is that ^R1 represents a _C1-8 alkyl group substituted by a substituent and the substituent contains a hydroxyl group, wherein each carbon atom of the _C1-8 alkyl group carries at least one hydrogen atom;

When ^R1 represents a _C1-8 alkyl group substituted by a substituent and the substituent contains a hydroxyl group, the five-membered ring does not represent an imidazole ring group;

^R1 represents a _C1-8 alkyl group substituted by a substituent, and the substituent contains an amino group, a _C1-8 alkylamino group, or a ( _C1-8 alkyl) _2amino group; at least one of X and Y represents a 6-14 alkyl aromatic group optionally substituted by a substituent.

When the five-membered ring is a thiophene ring group, m is not 0;

When a five-membered ring contains one nitrogen atom, the five-membered ring is bonded to the nitrogen atom through a non-nitrogen atom.

^R1 represents -(optionally substituted _C0-8 alkyl)C(O) ^NR4. When ^R5 is used, neither X nor Y is a bond, the five-membered ring represents a furan ring group, and m is not 0.

In another embodiment of the invention, a compound of general formula (II), a pharmaceutically acceptable salt thereof, an ester thereof, a prodrug thereof, a solvate thereof, a deuterated thereof, or a stereoisomer thereof is provided, which is a compound of general formula (III) below:

In another embodiment of the invention, compounds of general formula (II), pharmaceutically acceptable salts thereof, esters thereof, prodrugs thereof, solvates thereof, deuterated derivatives thereof, or stereoisomers thereof, are provided, which are compounds of general formula (IV) below:

In another embodiment of the invention, compounds of general formula (II), pharmaceutically acceptable salts thereof, esters thereof, prodrugs thereof, solvates thereof, deuterated derivatives thereof, or stereoisomers thereof are provided, which are compounds of general formula (V) or general formula (VI) below:

Where Z represents a _C1-8 alkyl group optionally substituted with ^R1 , and represents a single or double bond.

In another embodiment of the invention, the P ring in general formula (II) indicates that at least one double bond exists in the ring.

In another embodiment of the present invention, the P ring in general formula (II) is a heteroaromatic ring.

In another embodiment of the invention, the A ring in general formula (II) indicates that at least one double bond exists in the ring.

In another embodiment of the present invention, A in general formula (II) is a benzene ring or a heteroaromatic ring.

In another embodiment of the invention, W represents (optionally a _C0-8 alkyl group substituted with a substituent).

In another embodiment of the invention, each of the ^R1s in general formula (II) is independently selected from _C1-8 alkyl groups optionally substituted with substituents, -( _CH2 ) _O-4C ( ^R1a , ^R1b )( _CH2 ) _O- ^4OR3 , -( _CH2 ) _O-4C ( ^R1a , ^R1b )( _CH2 ) _O- ^{4NR3R3 ,} -( _CH2 ) _{O- 4C} (O) ^NR4R5 and -( _CH2 )O _- ^4S (O) _1-2R3 ^.

In another embodiment of the invention, ^R1 in general formula (II) is each independently selected from _C1-4 alkyl substituted with a substituent, -( _C1-4 alkyl substituted with a substituent) ^OR3 , -( _C1-4 alkyl substituted with a substituent) ^NR4R5 and -( _C0-4 alkyl substituted with ^a substituent)C(O) ^{NR4R5 and} -( _C1-4 alkyl substituted with a substituent)S( ^O ) _1-2R3 .

In another embodiment of the invention, ^R1 in general formula (II) is each independently selected from methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, isohydroxypropyl, hydroxybutyl, hydroxysec-butyl, hydroxytert-butyl, aminomethyl, aminoethyl, aminopropyl, isoaminopropyl, aminobutyl, aminosec-butyl, aminotert-butyl, -( _CH2 )O _-4C (O) _NH2 and -( _CH2 )O _{- 4S} (O) _2C1-4alkyl .

In another embodiment of the invention, ^R1a and ^R1b in general formula (II) are each independently selected from hydrogen atoms, amino groups, hydroxyl groups, and _C1-8 alkyl groups optionally substituted with substituents. Preferably, ^R1a and ^R1b in general formula (II) are each independently selected from hydrogen atoms, amino groups, hydroxyl groups, methyl groups, ethyl groups, propyl groups, isopropyl groups, butyl groups, sec-butyl groups, and tert-butyl groups.

In another embodiment of the invention, ^R3 , ^R4 and ^R5 in general formula (II) are each independently selected from hydrogen atoms and _C1-8 alkyl groups optionally substituted with substituents. Preferably, ^R3 , ^R4 and ^R5 in general formula (II) are each independently selected from hydrogen atoms, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl.

In another embodiment of the invention, R ^1′ in general formula (II) is selected from hydrogen atoms, hydroxyl groups, halogen atoms and _C1-8 alkyl groups.

In another embodiment of the invention, R1 ^′ in general formula (II) is connected to the carbon atom on the _C0-8 alkyl group of the W group, which is optionally substituted by a substituent, to form a 5-6 member unsaturated heterocyclic group together with the N and C(O) groups.

In another embodiment of the invention, R ^1′ in general formula (II) is selected from hydrogen atom, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl.

In another embodiment of the invention, R1 ^′ in general formula (II) is connected to the carbon atom on the _C0-8 alkyl group of the W group, which is optionally substituted by a substituent, to form a 6-membered unsaturated heterocyclic group together with the N and C(O) groups.

In another embodiment of the present invention, the group connected to the A ring and the W group in general formula (II) is a C atom.

In another embodiment of the present invention, ring A in general formula (II) is selected from benzene ring, pyridine ring, pyrimidine ring, pyridazine ring, pyrazine ring and triazine ring.

In another embodiment of the invention, in general formula (II), X and Y are each independently selected from bonds, optionally substituted benzene ring groups, optionally substituted pyrrole ring groups, optionally substituted pyrazole ring groups, optionally substituted 1,2,3-triazole ring groups, optionally substituted 1,2,4-triazole ring groups, optionally substituted tetraazole ring groups, optionally substituted furan ring groups, optionally substituted thiophene ring groups, optionally substituted oxazole ring groups, optionally substituted isoxazole ring groups, optionally substituted 1,2,4-oxadiazole ring groups, optionally substituted pyridine ring groups, optionally substituted indole ring groups, -(C＝C)-, -(C≡C)-, =N-, -C(O)-NR ^c - Optionally substituted group ...

In another embodiment of the invention, X and Y in general formula (II) are not simultaneously 6-14 membered aromatic groups that are optionally substituted with substituents. Preferably, X and Y in general formula (II) are not simultaneously benzene ring groups that are optionally substituted with substituents.

In another embodiment of the invention, in general formula (II), when one of X and Y represents a bonding bond, the other does not represent a 6-14 membered aromatic group optionally substituted by a substituent. Preferably, in general formula (II), when one of X and Y represents a bonding bond, the other does not represent a benzene ring group optionally substituted by a substituent.

In another embodiment of the invention, when one of X or Y represents a bonding bond, the other represents -(C≡C)-.

In another embodiment of the invention, in general formula (II), at least one of X and Y is -(C＝C)- or -(C≡C)-.

In another embodiment of the invention, neither X nor Y represents a bonding bond.

In another embodiment of the present invention, in the P ring of general formula (II), the C atom is attached to the X group.

In another embodiment of the invention, at least one of _P1 , _P2 , _P3 , _P4 and _P5 is a group selected from ^NRc , O and S.

In another embodiment of the invention, the P ring in general formula (II) is selected from pyrrolidine ring groups, pyrrolidine ring groups, imidazoline ring groups, imidazoline ring groups, pyrazole ring groups, pyrrole ring groups, pyrazole ring groups, 1,2,3-triazole ring groups, 1,2,4-triazole ring groups, thiophene ring groups, thiazole ring groups, isothiazole ring groups, 1,2,4-thiadiazole ring groups, oxazole ring groups, isoxazole ring groups, and 1,2,4-oxadiazole ring groups.

In another embodiment of the invention, the P ring in general formula (II) is selected from furan group, pyrrole group, pyrazole group, triazole group and thiophene group.

In another embodiment of the invention, each occurrence of B in general formula (II) is independently selected from a _C1-8 alkyl group that is bonded, optionally substituted, and at least one carbon atom is replaced by at _least one group from O and NR ^c , and optionally substituted.

In another embodiment of the invention, each of the Cs in general formula (II) independently represents a hydrogen atom, a _C1-8 alkyl group optionally substituted with a substituent, -OR ^c , -NR ^c R ^c , a 3-10 membered cycloalkyl group optionally substituted with a substituent, a 3-10 membered heterocyclic group optionally substituted with a substituent, a 5-10 membered heteroaryl group optionally substituted with a substituent, and a 6-10 membered aryl group optionally substituted with a substituent.

In another embodiment of the invention, each C in general formula (II) is independently selected from hydrogen atoms, optional _C1-4 alkyl groups substituted with substituents, -OR ^c , -NR ^c R ^c , optional 3-8 cyclic alkyl groups substituted with substituents, optional 3-8 heterocyclic groups substituted with substituents, optional 5-6 heteroaryl groups substituted with substituents, and optional 6-10 aryl groups substituted with substituents.

In another embodiment of the invention, C in general formula (II) may be any of the following groups that are optionally substituted by substituents:

Where Q is a group selected from CR ^{a Ra} , NR ^c , O and S.

In another embodiment of the invention, m in general formula (II) is 0, 1, 2 or 3.

In another embodiment of the invention, when ^R1 in general formula (II) represents -(optionally substituted _C0-8 alkyl)C( ^O ) ^NR4R5 , both X and Y represent -(C≡C)-, P represents a furan ring group, and m is not 0.

In another embodiment of the invention, ^R1 in general formula (II) represents a _C1-8 alkyl group substituted by a substituent, and the substituent contains an amino group, a _C1-8 alkylamino group, or a ( _C1-8 alkyl) _2amino group, at least one of X and Y represents a 6-14 nucleotide aromatic group optionally substituted by a substituent, and the P ring represents a pyrazole ring group.

In another embodiment of the invention, the P ring does not represent an imidazole ring group.

In another embodiment of the present invention, X is selected from -(C＝C)-, -(C≡C)-, 5-10-membered heteroaromatic groups optionally substituted by substituents, and 6-10-membered aromatic groups optionally substituted by substituents, and Y is selected from bonding bonds, -(C＝C)-, and -(C≡C)-, and when Y represents a bonding bond, X does not represent 6-10-membered aromatic groups optionally substituted by substituents.

In another embodiment of the invention, ^Ra in general formula (II) is either absent or, each time it appears, is independently selected from hydrogen, cyano, mercapto, halogen, carboxyl, nitro, -OR ^c , -NR ^c R ^c , -C(O)OR ^c , -S(O) _1-2 -OR ^c , optional substituted _C1-4 alkyl, -(optionally substituted _C1-4 alkyl)OR ^c , -(optionally substituted _C1-4 alkyl)NR ^c R ^c , optional substituted _C1-4 alkoxy, optional substituted _C2-4 alkenyl, optional substituted C2-4 alkynyl, optional substituted _3-10 membered cycloalkyl, optional substituted 3-10 membered heterocyclic, optional substituted 6-10 membered aryl, and optional substituted 5-10 membered heteroaryl.

In another embodiment of the invention, ^Ra in general formula (II) is either absent or, each time it appears, is independently selected from hydrogen atom, halogen atom, carboxyl group, nitro group, hydroxyl group, amino group, methyl group, ethyl group, propyl group, isopropyl group, butyl group, sec-butyl group, tert-butyl group, hydroxymethyl group, hydroxyethyl group, hydroxypropyl group, isohydroxypropyl group, hydroxybutyl group, hydroxysec-butyl group, hydroxytert-butyl group, aminomethyl group, aminoethyl group, aminopropyl group, isoaminopropyl group, aminobutyl group, aminosec-butyl group, aminotert-butyl group, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, sec-butoxy group, tert-butoxy group, optionally substituted 3-6 membered cycloalkyl group, optionally substituted 3-6 membered heterocyclic group, optionally substituted 6-10 membered aryl group and optionally substituted 5-6 membered heteroaryl group.

In another embodiment of the invention, ^Rc in general formula (II) is either absent, or, each time it appears, is independently selected from hydrogen atom, halogen atom, carboxyl group, optional substituent-substituted _C1-4 alkyl, -(optionally substituent-substituted _C1-4 alkyl) ^ORb , -(optionally substituent-substituted _C1-4 alkyl) ^NRbRb , ^optional substituent-substituted _C2-4 alkenyl, optional substituent-substituted _C2-4 alkynyl, optional substituent-substituted 3-10 membered cycloalkyl, optional substituent-substituted 3-10 membered heterocyclic group, optional substituent-substituted 6-10 membered aryl and optional substituent-substituted 5-10 membered heteroaryl, or, in the case where two ^Rc are attached to the same atom, the two ^Rc together with the attached atom form an optional substituent-substituted 3-10 membered heterocycle.

In another embodiment of the invention, ^Rc in general formula (II) is either absent, or each time it appears, it is independently selected from hydrogen atom, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, isohydroxypropyl, hydroxybutyl, hydroxysec-butyl, hydroxytert-butyl, aminomethyl, aminoethyl, aminopropyl, isoaminopropyl, aminobutyl, aminosec-butyl, aminotert-butyl, optional substituted 3-6 membered cycloalkyl, optional substituted 3-6 membered heterocyclic group, optional substituted 6-10 membered aryl and optional substituted 5-6 membered heteroaryl, or when two ^Rc are attached to the same atom, the two ^Rc together with the attached atom form an optional substituted 3-6 membered heterocycle.

In another embodiment of the invention, ^Rb in general formula (II) is each independently selected from hydrogen atom, halogen atom, carboxyl group, sulfonic acid group, optional substituted _C1-4 alkyl group, optional substituted _C1-4 alkylsulfonyl group, optional substituted 3-10 membered cycloalkyl group, optional substituted 3-10 membered heterocyclic group, optional substituted 6-10 membered aryl group, and optional substituted 5-10 membered heteroaryl group.

In another embodiment of the invention, ^Rb in general formula (II) is each independently selected from hydrogen atom, halogen atom, carboxyl group, methyl group, ethyl group, propyl group, isopropyl group, butyl group, sec-butyl group, tert-butyl group, 3-6 membered cycloalkyl group optionally substituted with a substituent, 3-6 membered heterocyclic group optionally substituted with a substituent, 6-10 membered aryl group optionally substituted with a substituent, and 5-6 membered heteroaryl group optionally substituted with a substituent.

In another embodiment of the invention, R ^d in general formula (II) is each independently selected from hydrogen atom, hydroxyl group, mercapto group, halogen atom, carboxyl group, nitro group, amino group, optional _C1-4 alkylamino group substituted with a substituent, (optionally _C1-4 alkyl) _2amino group substituted with a substituent, sulfonic acid group, optional _C1-4 alkyl group substituted with a substituent, optional _C1-4 alkoxy group substituted with a substituent, optional _C1-4 alkylsulfonyl group substituted with a substituent, optional 3-10 membered cycloalkyl group substituted with a substituent, optional 3-10 membered heterocyclic group substituted with a substituent, optional 6-10 membered aryl group substituted with a substituent, and optional 5-10 membered heteroaryl group substituted with a substituent.

In another embodiment of the invention, R ^d in general formula (II) is each independently selected from hydrogen atom, hydroxyl group, mercapto group, halogen atom, amino group, sulfonic acid group, methyl group, ethyl group, propyl group, isopropyl group, butyl group, sec-butyl group, tert-butyl group, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, sec-butoxy group, tert-butoxy group, 3-6 membered cycloalkyl group optionally substituted with a substituent, 3-6 membered heterocyclic group optionally substituted with a substituent, 6-10 membered aryl group optionally substituted with a substituent, and 5-6 membered heteroaryl group optionally substituted with a substituent.

In another embodiment of the invention, the substituents in "optionally substituted" are each independently selected from: hydroxyl, mercapto, carboxyl, cyano, nitro, amino, halogen atom, sulfonic acid group, _C1-4 alkyl, _C1-4 alkoxy- _C1-4 alkyl, hydroxy- _C1-4 alkyl, amino- _C1-4 alkyl, carboxyl- _C1-4 alkyl, ester- _C1-4 alkyl, _C1-4 alkoxy, hydroxy- _C1-4 alkoxy, amino- _C1-4 alkoxy, carboxyl _{-C1-4 alkoxy} , _C1-4 alkyl-C1-4 alkoxy, _C1-4 alkoxy- _C1-4 alkoxy, ester- _C1-4 alkoxy, _C1-4 alkylamino, ( _C1-4 alkyl) _2amino , amino- _C1-4 alkylamino, (amino- _C1-4 alkyl) _2amino , _C1-4 alkyl ester, aminocarbonyl, _C1-4 alkylaminocarbonyl, ( ... _1-4 alkyl) _2- aminocarbonyl, C1-4 alkylcarbonyl, _C1-4 alkylcarbonyloxy, _C1-4 alkylcarbonylamino, C1-4 alkylcarbonylamino, _C1-4 alkylsulfonylamino, halo _{- C1-4} alkyl, halo-C1-4 alkoxy, _C1-4 alkylsulfonyl, _C1-4 alkylsulfinyl, _C1-4 alkylthio, 3-10 membered cycloalkyl, _6-10 membered aryl, _3-10 membered heterocyclic, 5-10 membered heteroaryl and oxo.

In another embodiment of the invention, the substituents in "optionally substituted" are each independently selected from: hydroxyl, mercapto, carboxyl, cyano, nitro, amino, halogen atom, sulfonic acid group, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, isohydroxypropyl, hydroxybutyl, hydroxysec-butyl, hydroxytert-butyl, aminomethyl, aminoethyl, aminopropyl, isoaminopropyl, aminobutyl, aminosec-butyl, aminotert-butyl, aminotert-butyl Butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, tert-butoxy, halomethyl, haloethyl, halopropyl, haloisopropyl, halobutyl, halosec-butyl, halotert-butyl, halomethoxy, haloethoxy, halopropoxy, haloisopropoxy, halobutoxy, halosec-butoxy, halotert-butoxy, 3-6 membered cycloalkyl, 6-10 membered aryl, 3-6 membered heterocyclic, 5-6 membered heteroaryl and oxo.

Invention Effects

The compounds of this invention are inhibitors of UDP-3-O-(R-hydroxytetradecanoyl)-N-acetylglucosamine deacetylase (LpxC). These compounds exhibit excellent antibacterial activity, particularly against Gram-negative bacteria.

Furthermore, the compounds of the present invention have the following advantages compared to the closest prior art:

(1) The compounds of the present invention, their pharmaceutically acceptable salts, esters, their prodrugs, their solvates, deuterated derivatives or their stereoisomers have good anti-Gram-negative bacterial activity;

(2) The compounds of the present invention, their pharmaceutically acceptable salts, esters, their prodrugs, their solvates, deuterated derivatives or their stereoisomers exhibit good biological stability and excellent in vivo metabolic stability.

(3) The preparation process of the compounds of the present invention, their pharmaceutically acceptable salts, esters, their prodrugs, their solvates, deuterated products or their stereoisomers is simple and of stable quality.

Detailed Implementation

The embodiments of the present invention will be described in more detail below with reference to specific examples. However, those skilled in the art will understand that the specific embodiments described herein are for illustrative purposes only and should not be considered as limiting the scope of protection of the present invention. Rather, the present invention is intended to cover all alternatives, modifications, and equivalents that may be included within the scope of the present invention as defined by the claims.

Unless otherwise specified, the various embodiments of the present invention can be combined in any way, and the resulting transformations, modifications, and alterations of the technical solutions are also included within the scope of the present invention and do not exceed the scope of the present invention.

All publications, patent applications, patents, and other references mentioned in this specification are incorporated herein by reference. Unless otherwise defined, all technical and scientific terms used in this specification have the meanings commonly understood by those skilled in the art. In case of conflict, the definitions in this specification shall prevail.

In the context of this specification, except where expressly stated, any matters or issues not mentioned herein shall apply directly to those known in the art without any modification. Furthermore, any implementation described herein may be freely combined with one or more other implementations described herein, and any resulting technical solutions or concepts shall be considered part of the original disclosure or original record of this invention, and should not be regarded as new content not disclosed or anticipated herein, unless those skilled in the art consider such combination to be clearly unreasonable.

In this invention, the expression "C _ab group" (where a and b represent integers greater than or equal to 0, and a < b) indicates that the "group" has ab carbon atoms. For example, C _1-4 alkyl means an alkyl group with 1-4 carbon atoms, C _1-4 alkoxy means an alkoxy group with 1-4 carbon atoms, C _3-10 cycloalkyl means a cycloalkyl group with 3-10 carbon atoms, and C _1-4 alkoxy-C _1-4 alkyl means a group formed by the bonding of an alkoxy group with 1-4 carbon atoms and an alkyl group with 1-4 carbon atoms. When the subscript of C is 0, it means that the C ₀ group does not exist. For example, in "-C _a alkyl NH ₂ ", when a is 0, it means "-NH ₂ ".

In this invention, "(group) _ab " (where a and b represent integers greater than or equal to 0, and a < b) indicates the presence of ab groups. When the subscript is 0, it indicates the absence of that group. For example, -S(O) _0-2 indicates that 0, 1, or 2 oxygen atoms can be bonded to S, which are -S-, -S(O)-, and -S(O) _2- . For example, -( _CH2 ) _0-2 OH represents -OH, _-CH2OH , and -( _CH2 ) _2OH .

In this invention, "C( ^R1a , ^R1b )" indicates that ^R1a and ^R1b are each bonded to a carbon atom. For example, "-C(H, _CH3 )OH" indicates that the H atom and _-CH3 are each bonded to a carbon atom, which are functional groups.

In the following explanations, the meaning of groups with subscript 0 will not be explained; the explanation will begin with the meaning of subscript 1. For example, regarding the explanation of _C0-8 alkyl groups, since _C0 groups indicate that alkyl groups are not present, the explanation will begin with _C1 alkyl groups.

In this invention, "base" and "group" refer to monovalent groups or, as needed, divalent or higher groups that conform to the required valence. For example, "cycloalkyl" (also referred to as "cycloalkyl group") includes monovalent groups obtained by removing one hydrogen atom from a cycloalkane, and also includes divalent or higher groups obtained by removing two or more hydrogen atoms from the same carbon atom or two or more different carbon atoms of a cycloalkane. For example, when "cycloalkyl" is a terminal group, it is connected to other parts of the structural formula as a monovalent group when it does not carry substituents, and when it carries substituents, the cycloalkyl exhibits a corresponding valence number (number of substituents + 1) according to the number of substituents carried. Those skilled in the art can determine the valence number represented by "base" and "group" without any doubt.

The "halogen atom" mentioned in this invention refers to fluorine atoms, chlorine atoms, bromine atoms, iodine atoms, etc. Fluorine atoms, chlorine atoms, and bromine atoms are preferred.

The term "halogenation" as used in this invention refers to the substitution of one or more hydrogen atoms on any carbon atom in a substituent by one or more identical or different halogen atoms. It can be monohalogenation, polyhalogenation, or full halogenation, meaning that all positions in the halogen atom substituent group that can be substituted are included.

In this invention, " _C1-8 alkyl" refers to a straight-chain or branched monovalent or, as needed, divalent or higher alkyl group derived from a straight-chain or branched alkane containing 1-8 carbon atoms by removing one or more hydrogen atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1,2-dimethylpropyl, etc. " _C1-8 alkyl" can also be _C1-6 alkyl, _C1-4 alkyl, or _C1-3 alkyl. In this invention, "C _1-6 alkyl" refers to the above-mentioned examples containing 1-6 carbon atoms, "C _1-4 alkyl" refers to the above-mentioned examples containing 1-4 carbon atoms, and "C _1-3 alkyl" refers to the above-mentioned examples containing 1-3 carbon atoms.

The term "C _2-8 alkenyl" as used in this invention refers to a monovalent or, if desired, divalent or higher alkenyl group derived from a straight-chain or branched olefin containing 2 to 8 carbon atoms and at least one carbon-carbon double bond by removing one or more hydrogen atoms. Examples include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadien-1-yl, 1-penten-3-yl, 2-penten-1-yl, 3-penten-1-yl, 3-penten-2-yl, 1,3-pentadien-1-yl, 1,4-pentadien-3-yl, 1-hexen-3-yl, and 1,4-hexadien-1-yl. Preferably, the "C _2-8 alkenyl" group contains one carbon-carbon double bond. "C _2-4 alkenyl" refers to the above examples containing 2 to 4 carbon atoms.

The "C _2-8 ynyl" in this invention refers to a monovalent or, if necessary, divalent or higher ynyl group derived from a straight-chain or branched alkyne containing at least one carbon triple bond by removing one or more hydrogen atoms. Examples include ethynyl, propynyl, 2-butyn-1-yl, 2-pentyn-1-yl, 3-pentyn-1-yl, 4-methyl-2-pentyn-1-yl, 2-hexyn-1-yl, 3-hexyn-1-yl, and 3-hexyn-2-yl. Preferably, the "C _2-8 ynyl" contains one carbon triple bond. "C _2-4 ynyl" refers to the above examples containing 2-4 carbon atoms.

In this invention, "C _1-8 alkoxy" refers to the "C _1-8 alkyl" group defined above, which is a group connected to the parent compound via an oxygen atom, i.e., a "C _1-8 alkyl-O-" group. Examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentoxy, neopentoxy, n-hexoxy, and heptoxy. "C _1-4 alkoxy" refers to the above examples containing 1-4 carbon atoms, i.e., a "C _1-4 alkyl-O-" group.

In this invention, _the groups containing " _C1-8 alkyl,"" _C1-8 alkylamino,""( _C1-8 alkyl) _2amino ,"" _C1-8 alkyl ester,"" _C1-8 alkylaminocarbonyl," and "halogenated _C1-8 alkyl," etc., represent groups obtained by bonding the aforementioned " _C1-8 alkyl" with corresponding groups such as _{"C1-8 alkoxy} ,""amino,""ester,""aminocarbonyl," and "halogen atom." Similarly, the description of groups containing " _C1-4 alkyl" indicates groups obtained by bonding the aforementioned " _C1-4 alkyl" with corresponding groups. Likewise, groups containing " _C1-8 alkoxy" and " _C1-4 alkoxy" represent groups obtained by bonding the aforementioned " _C1-8 alkoxy" and " _C1-4 alkoxy" with corresponding groups.

In this invention, the cyclic group can be a monocyclic system or a polycyclic system. In the case of a polycyclic system, two or more rings are connected by bridging rings, spirocyclic rings, or fused rings. A bridging ring refers to a fused ring structure formed by two or more cyclic structures sharing two non-adjacent ring atoms. A spirocyclic ring refers to a fused ring structure formed by two or more cyclic structures sharing one ring atom. A fused ring refers to a fused ring structure formed by two or more cyclic structures sharing two adjacent ring atoms (i.e., sharing one bond).

The "3-12 membered cycloalkyl" in this invention refers to a saturated cyclic alkane group containing 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 cyclic carbon atoms. It can be a monovalent group or a divalent or higher group as needed. It can be a monocyclic system or a polycyclic system. The "3-12 membered cycloalkyl" in this invention can be a 3-12 membered cycloalkyl, a 3-10 membered cycloalkyl, a 3-8 membered cycloalkyl, or a 3-6 membered cycloalkyl. Examples of cycloalkyl groups in this invention include, but are not limited to: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopentane-1,3-diyl, cyclohexane-1,4-diyl, cycloheptane-1,4-diyl, norbornel, and adamantyl; monovalent or divalent groups derived from bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and bicyclo[4.2.1]nonane; monovalent or divalent groups derived from the following spirocycles by removing one hydrogen atom or, as needed, two or more hydrogen atoms: monovalent or divalent groups derived from the following bridged rings by removing one hydrogen atom or, as needed, two or more hydrogen atoms:

The "3-12 membered heterocycle" described in this invention refers to a non-aromatic cyclic hydrocarbon containing at least one (1-5, 1-4, 1-3, 1-2, or 1) group selected from O, S, and N as cyclic atoms within the ring, provided that the ring of said group does not contain two adjacent O or S atoms. It can be a heterocycle with 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 cyclic atoms. At least one double bond may be optionally present in the ring, thereby forming an unsaturated heterocyclic group. Furthermore, in the heterocycle of this invention, the cyclic atoms may optionally be oxidized, i.e., the C and S as cyclic atoms may form C(O), S(O), or S(O) _₂ groups. The heterocycle of this invention can be a monocyclic system or a polycyclic system. Examples of heterocycles include ethylene oxide, thiohexacyclopropane, oxetane, 1,2-dioxetane, thiohexacyclopropane, azirone, 1,2-diazacyclobutane, azirone butadiene, 1,2-diazacyclobutene, pyrrolidone (4,5-dihydropyrrole, 2,5-dihydropyrrole), pyrrolidine, imidazoline (4,5-dihydroimidazolium), imidazoline, pyrazolidine, piperidine, piperazine, morpholine, thiomorpholine, tetrahydropyran, dihydropyridine, dihydropyridazine, dioxane, oxathiapentyl ring, and sulfide ring. Pentane, tetrahydrofuran, tetrahydrothiazole, tetrahydroisothiazole, 2-pyridone, 4-pyridone, azacycloheptatriene, 1,2-diazacycloheptatriene, 1,3-diazacycloheptatriene, 1,4-diazacycloheptatriene, azacyclooctatetraene, 1,4-dihydro-1,4-diazacyclooctatetraene, 1,2-dithioheptatriene, 4,5-dihydrofuran, 2,5-dihydrofuran, 2,5-dihydrothiophene, 4,5-dihydrothiophene, 1,2-dithioheptatriene, 1,3-dithioheptatriene, 2H-pyran, 2H-pyran-2-one, 3... 4-Dihydro-2H-pyran, 4H-pyran, 4H-pyran-4-one, 1,4-dioxanediene, 1,4-dithiocyclohexanediene, 1,4-oxothiocyclohexanediene, oxacycloheptatriene, thiocycloheptatriene, 1,4-dioxane-octanetriene, 4,5-dihydrooxazole, 2,3-dihydrooxazole, 4,5-dihydroisoxazole, 2,3-dihydroisoxazole, 4,5-dihydrothiazole, 2,3-dihydrothiazole, 2H-1,2-oxazine, 4H-1,2-oxazine, 6H-1,2-oxazine, 2H-1,3-oxazine, 4H Monocyclic heterocycles such as -1,3-oxazine, 5,6-dihydro-4H-1,3-oxazine, 6H-1,3-oxazine, 2H-1,4-oxazine, 4H-1,4-oxazine, 2H-1,3-thiazine, 4H-1,3-thiazine, 5,6-dihydro-4H-1,3-thiazine, 6H-1,3-thiazine, 2H-1,4-thiazine, 4H-1,4-thiazine groups; fused heterocycles such as dihydroindole, isodihydroindole, benzopyran, benzodioxane, tetrahydroquinoline, benzo[d]oxazol-2(3H)-one, tetrahydrobenzothiophene. Furthermore, heterocycles obtained by replacing at least one ring carbon atom in the spirocyclic or bridged rings listed in the above examples of cycloalkyl groups with a heteroatom selected from O, S, and N can be cited.

The "3-12 membered heterocyclic group (heterocyclic group)" mentioned in this invention refers to a monovalent or divalent or higher group derived by removing one or more hydrogen atoms from any cyclic atom of the aforementioned "3-12 membered heterocycle". The "3-12 membered heterocyclic group" of this invention can be a 3-10 membered heterocyclic group, a 3-8 membered heterocyclic group, a 3-6 membered heterocyclic group, or a 5-6 membered heterocyclic group.

The "6-14 aryl (aromatic group)" described in this invention refers to a monovalent group having 6-14 cyclic carbon atoms, or a divalent or higher group as needed, derived from an aromatic carbocyclic hydrocarbon. Examples of 6-14 aryl groups include phenyl, naphthyl, phenanthryl, and anthraceneyl. Examples of 6-10 aryl groups include phenyl and naphthyl. When a divalent group is used, examples include phenylene and naphthylene.

The "5-14 membered heteroaryl (heteroaryl group)" described in this invention refers to an aromatic monovalent or, as needed, divalent or higher-valent cyclic hydrocarbon group containing at least one heteroatom selected from O, S, and N (which can be 1-5, 1-4, 1-3, 1-2, or 1) as a cyclic atom, provided that the ring of the group does not contain two adjacent O or S atoms. It can be a 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 membered heteroaryl ring group. Furthermore, in the heteroaryl group of this invention, the cyclic atom can optionally be oxidized without affecting the aromaticity; that is, the C and S atoms acting as cyclic atoms can form C(O), S(O), or S(O) ₂ groups. Additionally, the heteroaryl ring group of this invention can be a monocyclic system or a polycyclic system. Specifically, monocyclic heterocyclic groups such as pyrroleyl, pyrazinyl, pyrazolyl, indoleyl, tetrazolyl, furanyl, thiophenyl, pyridinyl, imidazolyl, 1,2,3-triazolyl, 1,2,4-triazolylcyclotyl, tetrazolyl, triazinyl, pyridazinyl, pyrimidinyl, pyrazinyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolyl (1,2,3-thiadiazole, 1,3,4-thiadiazole)yl, oxazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, and 1,2,4-oxadiazolyl can be listed; [further examples would follow] Fused heterocyclic groups of isoindolyl, indazole, indene, isodihydroindolyl, quinolinyl, isoquinolinyl, cenolinyl, 2,3-diazanaphthyl, quinazolinyl, naphthidyl, quinoxalinyl, purine, pteridinyl, benzimidazolyl, benzisoxazolyl, benzisoxadiazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, isobenzofuranyl, benzisoxazolyl, benzisoxazolyl, benzisoxazolyl, etc.

In this invention, "a _C1-8 alkyl whose at least one carbon atom is replaced by at least one group selected from S, O and NR ^c " means that at least one carbon atom in " _C1-8 alkyl" is replaced by at least one group selected from S, O and NR ^c . For example, the group that replaces the _C8 alkyl with the two groups O and ^NR _c can be C2alkyl- _OC2alkyl -NR ^c - _C2alkyl- .

The atoms in this invention include all isotopes of atoms. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and not limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of carbon include ^13C and ^14C . The isotope-labeled compounds of this invention can generally be prepared using conventional techniques known to those skilled in the art or by methods similar to those described herein, using suitable isotope-labeling reagents instead of the unlabeled reagents originally used.

In this invention, "heteroatom" refers to an atom selected from S, O, and N.

In this invention, the double bonds in the five-membered ring and the six-membered ring are randomly selected to exist in the ring. There can be one, two, or three double bonds, limited to the maximum number of double bonds that can exist in the ring. For example, in a five-membered ring, there can be one or two double bonds; in a six-membered ring, there can be one, two, or three double bonds.

In this invention, "optionally substituted by a substituent" means that it can be unsubstituted or substituted. In the case of substitution, it can be 1-substituted, 2-substituted, 3-substituted, 4-substituted, 5-substituted, 6-substituted, 7-substituted, 8-substituted, or more substitutions. In the case of multiple substitution (substituted by two or more substituents), the substituents can be the same or different from each other.

In one embodiment of the invention, the following compounds, pharmaceutically acceptable salts, esters, prodrugs, solvates, deuterated derivatives, or stereoisomers thereof are provided:

The term "pharmaceutically acceptable salt" for the compounds of this invention refers to a base addition salt or acid addition salt formed by the compounds of this invention with a pharmaceutically acceptable, non-toxic base or acid, including organic acid salts, inorganic acid salts, organic base salts, and inorganic base salts. Organic acid salts include formate, acetate, propionate, benzenesulfonate, benzoate, p-toluenesulfonate, 2,3-dihydroxysuccinate, camphorsulfonate, citrate, methanesulfonate, ethanesulfonate, propanesulfonate, fumarate, gluconate, glutamate, hydroxyethanesulfonate, lactate, maleate, malate, mandelate, mucilage, bis(hydroxynaphthyl)ate, pantothenate, succinate, tartrate, etc., with benzoate, benzenesulfonate, p-toluenesulfonate, methanesulfonate, citrate, maleate, fumarate, and tartrate being particularly preferred. Inorganic acid salts include hydrochloride, hydrobromide, hydroiodate, sulfate, phosphate, nitrate, etc., with hydrochloride, hydrobromide, sulfate, and phosphate being particularly preferred. Organic base salts include amine salts, including salts formed with primary, secondary and tertiary amines, cyclic amines and base ion exchange resins, and may be selected from salts formed with the following organic bases: for example, arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, meglumine, glucosamine, histidine, heparin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine and tromethamine, etc. Inorganic alkali salts include salts formed with ammonia, alkali metals, and alkaline earth metals, such as ammonium salts as well as lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, zinc salts, barium salts, aluminum salts, iron salts, copper salts, ferrous salts, manganese salts, and divalent manganese salts. Ammonium salts as well as sodium salts, potassium salts, calcium salts, and magnesium salts are particularly preferred.

The term "pharmaceutically acceptable ester" for the compounds of this invention refers to an ester that is hydrolyzed in vivo and includes esters that readily decompose in the human body, leaving behind the parent compound or its salts. Suitable ester groups include, for example, ester groups derived from pharmaceutically acceptable aliphatic carboxylic acids (especially alkanonic, alkenonic, cycloalkanonic, and alkanedioic acids), wherein each alkyl or alkenyl moiety preferably has six or fewer carbon atoms. Representative examples of specific esters include (but are not limited to) formate esters, acetate esters, propionate esters, butyrate esters, acrylate esters, and ethyl succinate esters.

The term "prodrug" for the compounds of this invention refers to a compound that, upon administration to a subject, undergoes a chemical transformation through metabolism or a chemical process to produce the compound of this invention and/or its salts and/or solvates. Any compound that is converted in vivo to provide a bioactive agent (i.e., the compound of this invention) is a prodrug within the scope and spirit of this invention. For example, compounds containing a carboxyl group can form physiologically hydrolyzable esters, which act as prodrugs by hydrolyzing in vivo to produce the compound of this invention itself. Such prodrugs are preferably administered orally because hydrolysis occurs primarily under the influence of digestive enzymes in many cases. Parenteral administration may be used if the ester itself is active or if hydrolysis occurs in the blood. For a detailed discussion of prodrugs, see T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14; and Edward B. Roche, ed., “Bioreversible Carriers in Drug Design,” American Pharmaceutical Association and Pergamon Press, 1987. Both of these references are incorporated herein by reference.

Additionally, the free form of the compounds of the present invention can be converted into the corresponding salt form, and vice versa. The free or salt form and/or solvated form of the compounds of the present invention can be converted into the non-solvated free or salt form of the corresponding compounds; and vice versa.

Some compounds of this invention contain one or more asymmetric centers, and thus can exist as racemic mixtures and racemic mixtures, single enantiomers, diastereomer mixtures, and single diastereomers. The compounds of this invention have asymmetric centers, each of which independently produces two optical isomers. The scope of this invention includes all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds. This invention includes all stereoisomers of these compounds. This invention includes cis and trans isomers. These stereoisomers are “R” or “S” configurations depending on the configuration of the substituents surrounding the chiral element. The terms “R” and “S” as used herein are configurations as defined in: IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem., 1976, 45: 13-30. The single stereoisomers of the compounds of the present invention can be prepared by synthesis from commercially available starting materials containing asymmetric or chiral centers, or by preparing racemic mixtures and then resolving them by methods well known to those skilled in the art. These resolving methods can be illustrated by the following examples: (1) linking a mixture of enantiomers with a chiral auxiliary agent, separating the resulting diastereomeric mixture by recrystallization or chromatography, and optionally releasing an optically pure product from the auxiliary agent, as described below: Furniss, Hannaford, Smith, and Tatchell, "Vogel's Textbook of Practical Organic Chemistry", 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England; or (2) directly separating the mixture of optically active enantiomers on a chiral chromatographic column; or (3) fractional recrystallization. Additionally, compounds with geometrical isomers of carbon-carbon double bonds and carbon-nitrogen double bonds are included in the compounds of the present invention. Substituents surrounding carbon-carbon or carbon-nitrogen double bonds are referred to as Z or E configurations, while substituents surrounding cycloalkyl or heterocyclic compounds are referred to as cis or trans configurations. All geometric isomers of the compounds of this invention and mixtures thereof are included within the scope of this invention.

The term "solvent" in the context of the present invention refers to a molecular complex comprising the compound of the present invention and one or more stoichiometric pharmaceutically acceptable solvent molecules, such as water and ethanol. When the solvent is water, it may also be referred to as a "hydrate".

The structures described in this invention also include compounds containing one or more isotopically enriched atoms. For example, compounds having the structures of this invention but comprising hydrogen replaced by deuterium or tritium or carbon replaced by carbon enriched with ^13C or ^14C are within the scope of this invention.

The compounds of this invention exhibit antibacterial activity, for example, against Gram-positive bacteria and Gram-negative bacteria. Specifically, the compounds of this invention exert antibacterial effects against Gram-positive or Gram-negative bacteria, inhibiting their reproduction or killing them. They may also inhibit bacterial reproduction while simultaneously killing some bacteria, reducing their numbers.

Examples of Gram-positive bacteria include Staphylococcus (Staphylococcus aureus, Staphylococcus epidermidis, etc.), Streptococcus (Streptococcus pyogenes, Group B Streptococcus, Pneumococcus, etc.), and Enterococcus (Enterococcus faecalis, Enterococcus faecium, etc.). Examples of Gram-negative bacteria include *Pseudomonas* (e.g., *Pseudomonas aeruginosa*), *Escherichia coli* (e.g., *E. coli*), *Klebsiella* (e.g., *Klebsiella pneumoniae*, *Klebsiella oxytoca*), *Haemophilus* (e.g., *Influenza*, *Paravirhabditis*), *Bordeix* (e.g., *Bordetella pertussis*, *Bacillus bronchiseptica*), *Serratia marcescens* (e.g., *Serratia marcescens*), *Proteus* (e.g., *Proteus mirabilis*), *Enterobacter* (e.g., *Enterobacter cloacae*), *Campylobacter jejuni*, *Citrobacter*, *Vibrio* (e.g., *Vibrio cholerae*), *Morganella* (e.g., *Morganella morganii*), and *Salmonella* (e.g., *Salmonella typhi*). The bacteria include Salmonella paratyphi, Shigella (dysentery bacteria, etc.), Acinetobacter (Acinetobacter baumannii, Acinetobacter calcoaceticus, etc.), Legionella (Legionella pneumophila, etc.), Bacteroides (Bacteroides fragilis, etc.), Neisseria (gonococci, cerebrospinal meningococci, etc.), Moraxella (Moraxella catarrhalis, etc.), Chlamydia (Chlamydia trachomatis, Chlamydia psittaci, etc.), and Helicobacter (Helicobacter pylori, etc.).

In another embodiment of the present invention, a pharmaceutical composition is provided comprising the compounds of the present invention.

In another embodiment of the present invention, a pharmaceutical composition comprising the compound of the present invention and one or more pharmaceutical excipients is provided.

The compounds of this invention can be combined with one or more pharmaceutical excipients to formulate any pharmaceutical preparation. Pharmaceutical excipients can include pharmaceutically acceptable carriers, excipients, and diluents. Examples of such carriers, excipients, and diluents include water, lactose, glucose, fructose, sucrose, sorbitol, mannitol, polyethylene glycol, and propylene glycol. Furthermore, commonly used thickeners, binders, disintegrants, pH adjusters, and solubilizers can be mixed into the aforementioned carriers, excipients, or diluents as needed. Using conventional formulation techniques, oral or non-oral medications such as tablets, pills, capsules, granules, powders, solutions, emulsions, suspensions, ointments, injections, and skin patches can be formulated.

The present invention also provides the use of the compounds of the present invention, or pharmaceutical compositions comprising the compounds of the present invention, for the treatment and/or prevention of infectious diseases.

One aspect of the present invention provides a method for inhibiting deacetylase (LpxC) in Gram-negative bacteria, thereby affecting bacterial growth, said method comprising administering the compound of the present invention to a patient who requires this inhibition.

In another aspect, the invention provides a method for inhibiting LpxC, thereby modulating the toxicity of bacterial infections, including administering the compound of the invention to a patient who requires such inhibition.

In some embodiments of the method for inhibiting LpxC using the compounds of the present invention, the MIC ₅₀ value of the compound as an LpxC inhibitor is less than or equal to 16 μg/mL. In other such embodiments, the MIC ₅₀ value is less than or equal to 8 μg/mL, less than or equal to 4 μg/mL, less than or equal to 2 μg/mL, less than or equal to 1 μg/mL, or less than or equal to 0.5 μg/mL.

Additionally, in one aspect of the invention, methods of treating and/or preventing disease in a subject include administering to the subject an effective antibacterial amount of the compound of the invention or the pharmaceutical composition of the invention. In a more preferred embodiment of the treatment method, the subject is a mammal. In some embodiments, the subject is a human.

In another aspect of the invention, a method for administering the compound of the invention to fermenting or non-fermenting Gram-negative bacteria is provided. In a preferred embodiment of the method for administering the compound of the invention to fermenting or non-fermenting Gram-negative bacteria, the Gram-negative bacteria are selected from species of Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, Alcaligenes xylosoxidans, Acinetobacter, Enterobacteriaceae, Haemophilus, and Neisseria.

In another embodiment of the invention, the invention provides a method for administering an inhibitory amount of the compound of the invention to Gram-negative bacteria, such as those selected from organisms, such as Serratia marcescens, Proteus, Klebsiella, Enterobacter, Citrobacter, Salmonella, Providencia, Morganella, Cedecea, Edwardsiella, and Enterobacteriaceae of Escherichia coli.

Another embodiment of the present invention provides a pharmaceutical composition comprising the compound of the present invention and at least one other pharmaceutically active ingredient.

In another embodiment of the present invention, a pharmaceutical composition comprising the compound of the present invention and at least one other pharmaceutically active ingredient is provided for the treatment and/or prevention of infectious diseases.

In another embodiment of the present invention, the active pharmaceutical ingredient is an antimicrobial agent other than the compound of the present invention (hereinafter also referred to as a second antimicrobial agent).

In another embodiment of the present invention, the active pharmaceutical ingredient is a non-antimicrobial agent other than the compound of the present invention.

The compounds of this invention are effective when used in combination with other pharmaceutically active ingredients. The compounds of this invention increase the sensitivity of Gram-negative bacteria to many existing types of antimicrobial agents. The combined use of the disclosed compounds with other antimicrobial agents is within the scope of this invention. Such antimicrobial agents include, but are not limited to, erythromycin, rifampin, nalidixic acid, carbenicillin, bacitracin, cycloserine, fosfomycin, vancomycin, piperacillin, amikacin, ciprofloxacin, polymyxin, ceftazidime, and imipenem.

A further aspect of the invention is the use of LpxC inhibitors for the treatment of infections, particularly bacterial infections. Treatment of bacterial infections with the compounds of the invention can be a primary infection or a co-infection caused by a bacterial species and one or more other infectious agents selected from bacteria, viruses, parasites, and fungi.

The term “treatment and/or prevention” as used in this invention refers to reversing, alleviating, suppressing or preventing a disease/condition, one or more symptoms of such a disease or condition.

The compounds of this invention can be used to treat and/or prevent diseases/symptoms/conditions caused by bacteria (especially Gram-negative bacteria) and bacteria that use LpxC in the biosynthesis of lipopolysaccharide (LPS) or endotoxins, which produce endotoxins.

The compounds of the present invention are useful in diseases/symptoms/conditions caused by bacterial production of lipid A and LPS or endotoxins, for example, for the treatment and/or prevention of sepsis, septic shock, systemic inflammation, local inflammation, chronic obstructive pulmonary disease (COPD), and acute exacerbations (AECB) of chronic bronchitis. For these conditions, treatment comprises administering the compounds of the present invention, the pharmaceutical compositions of the present invention, or the compounds of the present invention optionally in combination with other pharmaceutically active ingredients, wherein the other pharmaceutically active ingredients are secondary antimicrobial agents or non-antimicrobial agents.

For septicemia, septic shock, systemic inflammation, local inflammation, acute exacerbation of chronic obstructive pulmonary disease (COPD) and chronic bronchitis (AECB), preferred non-antimicrobial agents include anti-endotoxin and tyrosine kinase inhibitors containing endotoxin receptor-binding antibodies, endotoxin-binding antibodies, anti-CD14-binding protein antibodies, and anti-lipopolysaccharide-binding protein antibodies.

In the treatment of acute or chronic respiratory infections, the compounds of the present invention can also be used with non-antimicrobial agents via inhalation. Preferred non-antimicrobial agents in the above treatments include anti-inflammatory steroids, non-steroidal anti-inflammatory agents, bronchodilators, mucolytics, anti-asthmatic agents, and pulmonary fluid surfactants. In particular, non-antimicrobial agents may be selected from salbutamol, budesonide, clotrimazole, dexamethasone, nalorome, clotrimazole, fluticasone, flunisulfonamide, triamcinolone, ibuprofen, rofecoxib, naproxen, celecoxib, nalorome, ipratropium, osinurin, pibuterol, salmeterol, bronchiodilators, mucolytics, calfactants, beracontan, poractant alfa, surfaxin, and pulmozyme (also called alfa chain enzyme).

Therefore, the compounds of the present invention can be used alone or in combination with a second antimicrobial agent and/or non-antimicrobial agent to treat and/or prevent acute or chronic respiratory infections, including acute lung and hospital-acquired infections, such as those caused by *Enterobacter aerogenes*, *Enterobacter cloacae*, *Escherichia coli*, *Klebsiella pneumoniae*, *Klebsiella oxytoca*, *Proteus mirabilis*, and *Serratia marcescens*. s), Stenotrophomonas maltophilia, Pseudomonas aeruginosa, Burkholderia cepacia, Acinetobacter calcoaceticus, Alcaligenes xylosoxidans, Flavobacterium meningosepticum, Providencia stuartii, and Citrobacter freundii Infections caused by *Haemophilus influenzae*, common lung infections such as *Legionella* species, *Moraxella catarrhalis*, *Branhamella catarrhalis*, *Enterobacter* species, *Acinetobacter* species, *Klebsiella* species, and *Proteus* species. Infections caused by other bacteria such as Neisseria species, Shigella species, Salmonella species, Helicobacter pylori, Vibrionaceae, and Bordetella species, as well as Brucella species, Francisella tularensis, and/or Yersinia pestis.

When used to treat and/or prevent diseases caused by Gram-negative bacteria, the use of the compounds of the present invention can make Gram-negative bacteria sensitive to the effects of a second antimicrobial agent.

When the compounds of the present invention are used in combination with one or more second antimicrobial agents, the second antimicrobial agent includes, but is not limited to, the group consisting of:

(1) Macrolides or ketolides such as erythromycin, azithromycin, clarithromycin and telithromycin;

(2) β-lactams include penicillins, cephalosporins, and carbapenems such as carbapenems, imipenems, and meropenems;

(3) Single-strain antibiotics such as penicillin, penicillin V, methicillin, oxacillin, cloxacillin, dicloxacillin, nafcillin, ampicillin, amoxicillin, carbenicillin, ticarcillin, mezlocillin, piperacillin, azlocillin, temoxicillin, cefotaxime, cefepime, cefadroxil, cefadroxil, cefuroxime, cefprozil, cefaclor, chloramphenicol, cefoxitin, cefmetazole, cefotaxime, cefazolin, ceftriaxone, cefoperazone, ceftazidime, cefixime, cefpodoxime, cefbufen, cefdinir, cefpirome, cefepime, and amprolium;

(4) Quinolones such as nalidixic acid, oxoliquilic acid, norfloxacin, pefloxacin, enoxacin, ofloxacin, levofloxacin, ciprofloxacin, timafloxacin, lomefloxacin, fleroxacin, gapfloxacin, sparfloxacin, trovafloxacin, clinfloxacin, gatifloxacin, moxifloxacin, sitafloxacin, canenafloxacin, gemifloxacin, and pazufloxacin;

(5) Antibacterial sulfonamides and antibacterial sulfanilides, including para-aminobenzoic acid, sulfadiazine, sulfaisoxazole, sulfamethoxazole and N-phthalylsulfathiazole;

(6) Aminoglycosides such as streptomycin, neomycin, kanamycin, paromomycin, gentamicin, tobramycin, amikacin, netilmicin, spectinomycin, sisomicin, dibekacin and isopamicin;

(7) Tetracyclines such as tetracycline, chlortetracycline, demeclocycline, minocycline, oxytetracycline, methacycline and doxycycline;

(8) Rifamycins such as rifampin (also called rifampin), rifapentine, rifabutin, benzoxazine, rifampin and rifaximin;

(9) Lincosamides, such as lincomycin and clindamycin;

(10) Glycopeptides such as vancomycin and teicoplanin;

(11) Streptocinoids such as quinupristin and dalfopristin;

(12) Oxazolidinones, such as linezolid;

(13) Polymyxins, polymyxins and polymyxin E;

(14) Trimethoprim and bacitracin;

(15) Linezolid and other drugs for treating Gram-positive bacteria.

The second antimicrobial agent can be administered in combination with the compound of the present invention, either before, simultaneously with, or after the administration of the compound of the present invention. When it is desired that the compound of the present invention and the second agent be administered simultaneously via the same route of administration, the compound of the present invention can be formulated into the same dosage form as the second antimicrobial agent. Examples of dosage forms containing the compound of the present invention and the second antimicrobial agent are tablets or capsules.

When used to treat acute or chronic respiratory infections, the compounds of the present invention may be used alone or in combination with a second antimicrobial agent administered by inhalation. In the inhalation case, preferred second antimicrobial agents are selected from tobramycin, gentamicin, aztreonam, ciprofloxacin, polymyxin B, phytohexidine, azithromycin, and polymyxin E.

The pharmaceutical compositions of the present invention can be formulated into any clinically or pharmaceutically acceptable dosage form, preferably oral formulations and injections.

The compounds of the present invention can be formulated into any pharmaceutically acceptable dosage form, which can be administered to mammals, such as humans, via oral, parenteral (intravenous, intramuscular, subcutaneous, or rectal) or local administration.

When used for parenteral administration, the compounds of the present invention can be formulated into injections, including sterile solution, emulsion, dispersion or suspension preparations for intramuscular injection, intravenous injection, intravenous drip, subcutaneous injection, etc., as well as sterile powder or concentrated solution for injection for preparation or dilution into solution, dispersion or suspension before use.

The injectable preparation can be produced using conventional methods in the existing pharmaceutical field, and can be produced using either aqueous or non-aqueous solvents. The most commonly used aqueous solvent is water for injection, but sodium chloride solution or other suitable aqueous solutions can also be used. Commonly used non-aqueous solvents are vegetable oils, such as soybean oil for injection, and other aqueous solutions of ethanol, propylene glycol, polyethylene glycol, etc. When preparing the injectable preparation, additives may be omitted, or suitable additives may be added according to the properties of the drug, such as osmotic pressure regulators, pH adjusters, solubilizers, fillers, antioxidants, antibacterial agents, emulsifiers, suspending agents, etc. Commonly used osmotic pressure regulators include sodium chloride, glucose, potassium chloride, magnesium chloride, calcium chloride, and sorbitol, with sodium chloride or glucose being preferred; commonly used pH regulators include acetic acid-sodium acetate, lactic acid, citric acid-sodium citrate, and sodium bicarbonate-sodium carbonate; commonly used solubilizers include polysorbate 80, propylene glycol, lecithin, polyoxyethylene castor oil, and cyclodextrin; commonly used fillers include lactose, mannitol, sorbitol, and dextran; commonly used antioxidants include sodium sulfite, sodium bisulfite, and sodium metabisulfite; and commonly used antibacterial agents include phenol, cresol, and chlorobutanol.

The pharmaceutical composition can also be formulated into dosage forms for rectal or local administration using conventional methods, including suppositories, ointments, creams, patches, powders, sprays, inhalers, etc.

When used for oral administration, the compounds of this invention can be formulated into conventional solid dosage forms, such as tablets, capsules, pills, and granules, using conventional methods; they can also be formulated into oral liquid dosage forms, such as oral solutions, oral suspensions, and syrups. Tablets are primarily ordinary oral tablets, but also include lozenges, sublingual tablets, oral patches, chewable tablets, dispersible tablets, soluble tablets, effervescent tablets, sustained-release tablets, controlled-release tablets, and enteric-coated tablets. Capsules, based on their solubility and release characteristics, can be classified into hard capsules, soft capsules, sustained-release capsules, controlled-release capsules, and enteric-coated capsules. Pills include drop pills, sugar pills, and small pills. Granules can be classified into soluble granules, suspension granules, effervescent granules, enteric-coated granules, sustained-release granules, and controlled-release granules.

In the preparation of oral formulations, suitable fillers, binders, disintegrants, lubricants, etc., can be added. Commonly used fillers include starch, powdered sugar, calcium phosphate, calcium sulfate dihydrate, dextrin, microcrystalline cellulose, lactose, pregelatinized starch, mannitol, etc.; commonly used binders include sodium carboxymethyl cellulose, PVP-K30, hydroxypropyl cellulose, starch paste, methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, gelatinized starch, etc.; commonly used disintegrants include dry starch, crospovidone, crospovidone carboxymethyl cellulose sodium, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, etc.; commonly used lubricants include magnesium stearate, talc, sodium lauryl sulfate, micronized silica gel, etc.

When the compounds of the present invention are formulated into pharmaceutical compositions (e.g., pharmaceutical preparations), the pharmaceutical compositions of the present invention contain 0.01-1000 mg of the compounds of the present invention, preferably 0.1-500 mg, more preferably 0.1-200 mg, and more preferably 1-100 mg, for example 0.5 mg, 1 mg, 3 mg, 5 mg, 10 mg, or 15 mg. The pharmaceutical compositions of the present invention (e.g., pharmaceutical preparations) may be in unit dose form, with each unit dose containing 0.01-1000 mg of the compounds of the present invention, preferably 0.1-500 mg, more preferably 0.1-200 mg, and more preferably 1-100 mg.

On the other hand, the present invention also provides the use of the compounds of the present invention in the preparation of medicaments for treating and/or preventing infectious diseases.

For adult patients, the compounds of the present invention can be administered in the above-described dosage form, once daily or divided into several doses, with a total dose of 0.001–1500 mg/day, preferably 0.01–1000 mg/day, more preferably 0.1–800 mg/day, and particularly preferably 1–600 mg/day, for example 250 mg/day, 400 mg/day, 500 mg/day, or 600 mg/day. It should be noted that the dosage of the compounds of the present invention can be appropriately increased or decreased according to the type of disease being treated, the patient's age, weight, symptoms, etc.

The compounds of this invention, as antibiotics, exhibit good antibacterial activity and good biological stability, especially against Gram-negative bacteria, and can be used to treat and/or prevent various diseases caused by Gram-negative bacteria.

Example:

The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following embodiments. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention.

First, the beneficial effects of the compounds of the present invention are further illustrated through antibacterial activity experiments, but this should not be construed as the compounds of the present invention having only the following beneficial effects.

Experimental Example 1: In vitro antibacterial activity of the compounds of the present invention

Test strains: The following clinical isolates were all purchased from public institutions.

Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853, Klebsiella pneumoniae ATCC700603, etc.

Test samples: Imipenem and ceftazidime: commercially available products. The following tests were conducted according to the content of the active ingredient in each commercially available product.

The chemical names and preparation methods of the compounds of this invention are given in the preparation examples of each compound.

Experimental method: Agar dilution method, referring to National Committee for Clinical Laboratory Standards. 2006. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard—Seventh Edition M7-A7, Vol. 26, No. 2, 2006, to determine the MIC of the compound. MIC represents the minimum inhibitory concentration.

Experimental results and conclusions:

Table 1. MIC (μg/mL) of some compounds of the present invention against Klebsiella pneumoniae

Table 2. MIC (μg/mL) of some compounds of the present invention against Escherichia coli

Table 3. MIC (μg/mL) of antibacterial activity against Pseudomonas aeruginosa for some of the compounds of this invention.

Experimental results show that the compounds of this invention exhibit excellent antibacterial activity against the tested strains. Specifically, the antibacterial activity of the compounds of this invention is superior to or comparable to that of the control compounds. Therefore, the compounds of this invention have good potential for clinical application.

Experimental Example 2: Evaluation of the in vitro antibacterial activity of the compounds of the present invention against polymyxin-resistant mcr-1 positive Enterobacteriaceae.

Test sample: The compound of the present invention was prepared according to the method of the preparation examples;

Polymyxin E sulfate, ampicillin, cefotaxime, gentamicin, ciprofloxacin, tetracycline, and trimethoprim-sulfamethoxazole were all commercially available products, and the following experiments were conducted according to the content of the active ingredient in each commercially available product.

The tested bacterial strains, including the *Escherichia coli* carrying the mcr-1 gene isolated from the animal and the quality control strain *Escherichia coli* ATCC25922, were provided by South China Agricultural University.

Experimental methods

The MIC values of the test drug against polymyxin-resistant mcr-1 positive Escherichia coli were determined using the agar dilution method in CLSI.

Preparation of drug-containing agar plates: Prepare the highest concentration of drug solution, and dilute the highest concentration of each compound solution by a factor of two. Take 1 mL of each dilution gradient drug solution (20× drug working solution) and add it to 19 mL of MH agar at 45℃ to 50℃. Immediately mix the agar and drug solution to prepare drug-containing agar plates with final concentrations of 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 and 0.06 μg/mL. Let them solidify and set aside for use. At the same time, a growth control group is set up.

Inoculum preparation and cultivation: After revitalizing the bacterial strain stored at -80℃, the turbidity was prepared with broth to reach or exceed 0.5 McFarland units. A 96-well plate was used as the inoculation tank. The bacterial suspension was diluted with sodium chloride injection to ensure a bacterial concentration of ^10⁴ CFU/point on the agar. The bacterial suspension was inoculated onto the marked drug-containing agar surface using a multi-point inoculation device. After the moisture at the inoculation points was completely absorbed by the agar, the plates were inverted and incubated at 37℃ for 16-20 hours.

Experimental results:

This experiment evaluated polymyxin-resistant mcr-1 positive Enterobacteriaceae. The in vitro MIC ₅₀ and MIC ₉₀ values of each compound are shown in Table 4.

Table 4. In vitro antibacterial activity of the compounds of this invention against polymyxin-resistant mcr-1 positive Enterobacteriaceae (unit: μg/mL)

Experimental conclusion:

As shown in Table 4, compounds 5 and 6 of this invention exhibit good in vitro antibacterial activity against mcr-1-positive Enterobacteriaceae resistant to polymyxin E sulfate, with MIC ₅₀ values of 0.125 μg/mL and 0.5 μg/mL, and MIC ₉₀ values of 0.5 μg/mL and 1 μg/mL, respectively. These values are superior to those of marketed drugs ampicillin, cefotaxime, gentamicin, ciprofloxacin, tetracycline, imipenem, and trimethoprim-sulfamethoxazole (≥4 indicates resistance). This demonstrates that the compounds of this invention have good potential for clinical application compared to existing compounds.

Experimental Example 3: Evaluation of the in vitro antibacterial activity of the compounds of the present invention against Pseudomonas aeruginosa.

Test sample: The compound of the present invention was prepared according to the method of the preparation example; compound C was prepared according to patent WO2008154642 A2.

Experimental methods:

The minimum inhibitory concentration (MIC) of the compound against the strain was determined according to the agar dilution method in the 2018 Clinical Laboratory Standards Institute (CLSI) guidelines.

Preparation of drug-containing petri dishes: (1) Prepare the highest concentration of the drug solution, with the highest concentration of each compound being 160 μg/mL. (2) Dilute the highest concentration of each compound solution by a factor of two to obtain a total of 10 concentration gradients. Take 2 mL of each dilution and add it to 18 mL of MH agar solution that has been autoclaved at 121℃ and cooled to about 50℃. Mix well and allow to stand and cool to obtain the drug-containing petri dishes, and label them accordingly. The final concentrations of the compounds are 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.06, and 0.03 μg/mL.

Inoculum preparation and inoculation: Select fresh single colonies, dilute them with 0.9% sodium chloride injection and adjust to 0.5# McFarland turbidity, then dilute them 10 times with 0.9% sodium chloride injection and inoculate them onto the marked drug-containing plates using a multi-point automatic inoculation instrument.

Experimental results:

This experiment evaluated the in vitro antibacterial activity of compound 6 and control compound C against Pseudomonas aeruginosa. Their MIC ₅₀ and MIC ₉₀ values are shown in Table 5.

Table 5. In vitro antibacterial activity of the compounds of this invention against Pseudomonas aeruginosa

Experimental conclusion:

As shown in Table 5, compound 6 of this invention exhibits excellent in vitro antibacterial activity against *Pseudomonas aeruginosa*, with MIC ₅₀ and MIC ₉₀ values of 0.5 μg/mL and 1 μg/mL, respectively. In contrast, compound C shows poor in vitro antibacterial activity against *Pseudomonas aeruginosa*, with MIC ₅₀ and MIC ₉₀ values of 4 μg/mL and >16 μg/mL, respectively. Overall, the compounds of this invention demonstrate superior antibacterial activity against *Pseudomonas aeruginosa* compared to compound C.

Experimental Example 4: Pharmacokinetic (PK) Determination

1. Experimental Design

2. Test sample

The compound of this invention was prepared in-house and dissolved in a suitable solvent.

3. Equipment

Instrumentation: API4000 LC-MS/MS

Column: Waters XBridge C ₁₈ (2.1×50mm, 5μm)

4. Blood collection

Rat blood collection: Animals were immobilized. Ten minutes prior to each time point, the tail was heated in a water bath, and approximately 200 μl of blood was collected via the tail vein. The collected blood was placed in tubes containing heparin sodium anticoagulant. Blood samples were centrifuged at 8000 rpm for 6 minutes at 4°C to obtain plasma samples. Plasma samples must be prepared within 30 minutes of blood collection. Plasma was stored at -80°C before testing.

5. Experimental Methods

(1) Take the sample to be tested out of the refrigerator (-80℃), let it melt naturally at room temperature, and then vortex for 5 minutes;

(2) Accurately transfer 20 μl of sample into a 1.5 ml centrifuge tube;

(3) Add 200 μl of internal standard solution;

(4) Vortex for 5 minutes, then centrifuge for 5 minutes (12,000 rpm);

(5) Accurately transfer 100 μl of supernatant to 100 μl of water, vortex for 5 min, and analyze by LC-MS/MS.

6. Data Processing Methods

The concentration of the test substance (plasma sample) was calculated using Analyst 1.6.1 from AB. Parameters such as mean, standard deviation, and coefficient of variation were calculated using Microsoft Excel (parameters directly output by Analyst 1.6.1 did not require calculation). PK parameters were calculated using Phrasight Phoenix 6.1 software.

Experimental results:

Table 6. PK results of the compounds of this invention in SD rats (IV 2 mg/kg)

Conclusion: Experimental results show that the compounds of this invention possess excellent in vivo metabolic stability, indicating their good drug-likeness. Therefore, the compounds of this invention have better prospects for clinical application.

The abbreviations and English expressions used in the embodiments have the following meanings:

TCL Thin-Layer Chromatography

Bu butyl

Et Ethyl

Me Methyl

EDCI 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

Hexane

THF (Tetrahydrofuran)

DMF (Dimethylformamide)

EA (ethyl acetate)

PE (petroleum ether)

DIEA N,N-Diisopropylethylamine

DCM (Dichloromethane)

NBS (N-bromosuccinimide)

TMS (Trimethylsilyl)

Boc tert-Butoxycarbonyl

TFA (Trifluoroacetic acid)

HOBt (Hydroxybenzotriazole)

HATU O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea

NH ₂ OTHP O-(tetrahydro-2H-pyran-2-yl)hydroxylamine

DMDMA N,N-Dimethylformamide dimethyl acetal

Pd(dppf) _Cl₂ [1,1′-bis(diphenylphosphine)ferrocene]palladium(II) dichloride

Example 1 4-((1E,3E)-4-(furan-2-yl)but-1,3-dienyl)-N-((2S,3R)-3-hydroxy-1- Preparation of (hydroxyamino)-1-oxobutane-2-yl)benzamide (compound 1)

(1) Preparation of intermediate 1

In a 250 mL three-necked flask, methyl p-bromomethylbenzoate (50 g, 0.22 mol) and triethyl phosphite (73 g, 0.45 mol) were added. The mixture was heated gradually to reflux under nitrogen protection. TLC monitoring showed the starting material disappearing (developing solvent: EA:PET = 1:10). Post-treatment: Under nitrogen protection, the mixture was cooled to room temperature, concentrated under reduced pressure to remove most of the remaining triethyl phosphite, and 100 mL of toluene was added for further concentration. This concentration was repeated three times until dry to obtain 68 g of a pale yellow oil (weight greater than the theoretical amount). This oil was used directly in the next reaction.

(2) Preparation of intermediate 2

Furanaldehyde (150 g, 1.56 mol) was added to a 5.0 L three-necked flask, followed by 900 mL of ethanol. The flask was cooled to below 10 °C in an ice-water bath. A solution of sodium hydroxide (40 g, 1.0 mol) dissolved in 1.8 L of purified water was added dropwise. After stirring for 30 minutes, an aqueous solution of acetaldehyde (400 g, 3.6 mol, 40% aqueous solution) was added dropwise, maintaining the system temperature at approximately 0 °C throughout the addition, which was completed in about 5 hours. After adding the acetaldehyde aqueous solution, the mixture was allowed to heat to room temperature overnight. Post-treatment: 2 L of methyl tert-butyl ether was added to the reaction system, and the organic phase was separated by stirring. The mixture was washed once with brine, concentrated, and a brown oily substance was obtained. This oily substance was then purified by column chromatography using petroleum ether as the eluent and silica gel. 56 g of the purified product was collected and directly used for the next reaction.

(3) Preparation of intermediate 3

Under nitrogen protection, intermediate 1 (56 g, 0.196 mol) was dissolved in 560 mL of dry THF. The liquid nitrogen-ethanol system was cooled to below -70°C. An 81 mL solution of n-butyllithium (2.5 M in Hexane) was added dropwise. After the addition was complete, the reaction was maintained at this temperature for 30 minutes. Intermediate 2 (22.6 g, 0.185 mol) was dissolved in 100 mL of dry THF. The temperature was controlled below -70°C, and the solution was added dropwise to the above solution. After the addition was complete, the reaction was maintained at this temperature for 2 hours. The temperature was then automatically raised to room temperature for post-treatment. The reaction system was slowly added to 400 ml of ice water, stirred, and the organic phase was separated. The aqueous phase was extracted twice with ethyl acetate. The organic phases were combined, washed twice with brine, dried with anhydrous sodium sulfate, concentrated and dried quickly, methyl tert-butyl ether was added, and the mixture was stirred overnight. The mixture was filtered to obtain a yellow solid with a wet weight of 30 g. The solid was added to 150 mL of THF and heated to 50 °C to dissolve, resulting in a clear yellow solution. The solution was hot filtered, cooled to precipitate the solid, filtered to obtain a yellow aliquot solid, and dried under vacuum to obtain an intermediate of 326 g, which was then directly used for the next reaction.

(4) Preparation of intermediate 1-1

Intermediate 3 (26g, crude product) was added to 260mL THF, and 12g of sodium hydroxide solution was dissolved in 130mL purified water. The mixture was heated to 45-50℃ and reacted overnight. The next day, TLC showed that a small amount of raw material had not reacted completely. The mixture was cooled to room temperature and filtered to obtain a yellow solid. The solid was added to a mixed solution of 150mL THF and 150mL ethyl acetate, and 1N hydrochloric acid solution was added dropwise until the pH of the system was about 1.0. The organic phase was separated, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was added to methyl tert-butyl ether and stirred for 2 hours, filtered, and dried under vacuum to obtain 15g of solid intermediate 1-1.

(5) Preparation of intermediate 1-2

Intermediate 1-1 (961 mg, 4 mmol) and L-threonine methyl ester hydrochloride (678 mg, 4 mmol) were dissolved in 8 mL of DMF. DIEA (1.034 g, 8 mmol) was added, and the solution was stirred for 50 min. After the solution became clear, HOBt (541 mg, 4 mmol) and EDCI (767 mg, 4 mmol) were added, and the reaction was stirred for 15 hours. The post-processed system was poured into water, filtered, the filter cake was washed with water, and dried to obtain 600 mg of crude intermediate 1-3.

(6) Preparation of Compound 1

The crude intermediate 1-3 (270 mg) was dissolved in 3 mL of methanol, and 1 mL of 50% hydroxylamine aqueous solution was added. The reaction was stirred for 40 hours. The post-treatment system was poured into water, filtered, the filter cake was washed with water and dried to obtain 79 mg of compound 1.

_Molecular formula: _{C19H20N2O5 Molecular} weight: _356.37 Mass spectrometry: (M+H): 357.1

^1H -NMR(d ₆ -DMSO, 800MHz) δ 10.66 (1H, s), 8.82 (1H, s), 7.98 (1H, d), 7.87 (2H, d), 7.69 (1H, s), 7.58 (2H, d), 7.23-7.16 (1H , m), 6.89-6.76 (2H, m), 6.69-6.66 (1H, m), 6.56 (2H, d), 5.1-4.8 (1H, m), 4.27 (1H, s), 4.03 (1H, s), 1.08 (3H, d).

Referring to the above experimental example, compound 25 was synthesized using racemic threonine methyl ester hydrochloride.

Example 2 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((4-(5-(hydroxymethyl) Preparation of (2-yl)furan-2-yl)phenyl)acetylene)benzamide (compound 2)

Compound 2 was synthesized according to Example 1.

_{Molecular formula: C₂₄H₂₂N₂O₆ Molecular weight} : _434.44 Mass spectrometry (M⁺): 434.15

^1H -NMR(d ₆ -DMSO, 800MHz) δ 10.68 (1H, s), 8.87 (1H, s), 7.96 (2H, d), 7.75 (2H, d), 7.67 (2H, d), 7.63 (2H, d), 7.73 (2H, d) ), 6.45(1H, d), 5.33(1H, t), 4.92(1H, brs), 4.47(2H, d), 4.28-4.25(1H, m), 4.03-4.01(1H, m), 1.10(3H, d).

Example 3 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((1E,3E)-4- Preparation of (5-(hydroxymethyl)furan-2-yl)but-1,3-dienyl)benzamide (compound 3)

(1) Preparation of intermediate 3-2

In a 250 mL three-necked flask, methyl p-bromomethylbenzoate (60 g, 0.26 mol, 1.0 eq.) and triethyl phosphite (87 g, 0.52 mol, 2.0 eq.) were added. The mixture was heated gradually to reflux under nitrogen protection. TLC monitoring continued until reactant 1 disappeared. The mixture was then cooled to room temperature under nitrogen protection, concentrated under reduced pressure to remove most of the remaining triethyl phosphite, and 100 mL of toluene was added for further concentration. This process was repeated three times until dry to obtain 78 g of a pale yellow oil. Intermediate 3-2 of this oil was used directly in the next reaction step.

(2) Preparation of intermediate 3-4

1L of toluene was added to a 2L three-necked flask, followed by starting material 3-3 (156g, 0.93mol, 1.0eq.). The mixture was heated under reflux for two days, then the reaction was terminated and cooled to room temperature. The system was filtered, and the solid was washed with methyl tert-butyl ether. After drying, 172g of intermediate 3-4 solid was obtained, with a yield of 33%, which was used directly in the next reaction.

(3) Preparation of intermediate 3-6

Under nitrogen protection, starting material 3-5 (185 g, 1.49 mol, 1.0 eq.) was dissolved in 560 mL of dry acetonitrile, and powdered sodium hydroxide (56 g, 0.196 mol, 1.05 eq.) was added. The mixture was then cooled in a water bath. 3-Bromopropene (450 g, 3.74 mol, 2.5 eq.) was added dropwise to the system. After the addition was complete, the mixture was stirred at room temperature. TLC (EA/PE ~ 1:5) showed that the reaction was complete. The reaction was then terminated, and the system was quenched in 1 L of water. The mixture was extracted with methyl tert-butyl ether (300 mL * 3), washed with water (300 mL * 2), washed with brine (300 mL * 1), dried, concentrated, and purified by column chromatography to obtain 16.8 g of intermediate 3-6, which was used directly in the next step.

(4) Preparation of intermediate 3-7

Intermediate 3-6 (14.8 g, 90.1 mmol, 1.0 eq.) and intermediate 3-4 (58.0 g, 135.2 mmol, 1.5 eq.) were added to 90 mL of DMF and stirred at room temperature. Sodium methoxide (14.6 g, 270.4 mmol, 3.0 eq.) was dissolved in 45 mL of methanol and added dropwise to the system, causing the temperature to rise. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. TLC (EA/PE ~ 1:5) showed that the reaction was complete. The reaction was then terminated, and the system was quenched in 200 mL of water. The mixture was extracted with ethyl acetate (100 mL * 3), washed with water (100 mL * 2), washed with brine (100 mL * 1), and dried and concentrated to obtain a brown solid-oil mixture.

The mixture was dissolved in 75 ml of tetrahydrofuran, and 75 ml of water and 35 ml of concentrated hydrochloric acid were added. The reaction was terminated at room temperature for 1 h. The system was quenched with 200 ml of water, extracted with ethyl acetate (100 ml * 3), washed with water (100 ml * 2), washed with brine (100 ml * 1), and dried and concentrated to obtain a brown solid-oil mixture. 100 ml of methyl tert-butyl ether was added and stirred for 0.5 h. The solid was filtered off, the mother liquor was concentrated, and purified by column chromatography to obtain 12.5 g of intermediate 3-7, which was used directly in the next step.

(5) Preparation of intermediate 3-8

Intermediate 3-2 (19.3 g, 65.1 mmol, 1.2 eq) was added to 200 mL of tetrahydrofuran. Under nitrogen protection, the temperature was lowered to -70 °C to -80 °C, and n-butyllithium (27.0 mL, 65.1 mmol, 1.2 eq) was added dropwise. The reaction was carried out at -70 °C to -80 °C for 30 min. Intermediate 3-7 (10.8 g, 56.2 mmol, 1.0 eq) was dissolved in 50 mL of tetrahydrofuran and added dropwise to the system. After the addition was complete, the mixture was allowed to return to room temperature and reacted overnight. TLC (EA/PE ~ 1:5) showed the reaction was complete. The reaction was terminated, and the system was quenched in 400 ml of saturated ammonium chloride solution. The mixture was extracted with ethyl acetate (100 ml * 3), washed with water (100 ml * 2), washed with brine (100 ml * 1), dried and concentrated until about 30 ml of solvent remained. 200 ml of petroleum ether was added, and the mixture was stirred at room temperature for 1 hour. After filtration and drying, 12 g of intermediate 3-8 was obtained, which was used directly in the next step.

(6) Preparation of intermediate 3-9

Intermediate 3-8 (12.8 g, 39.5 mmol, 1.0 eq), tetrakis(triphenylphosphine)palladium (4.6 g, 3.95 mmol, 0.1 eq), and ammonium acetate (15.2 g, 197.3 mmol, 5.0 eq) were added to 250 mL of acetonitrile. The mixture was deoxygenated under nitrogen and heated to 70 °C overnight. TLC (EA/PE ~ 1:3) showed complete reaction. The reaction was terminated, and the mixture was quenched in 500 mL of water. The mixture was extracted with ethyl acetate (300 mL x 2), washed with water (300 mL x 2), washed with brine (300 mL x 1), dried, passed through a silica gel pad, concentrated, and the residue was refluxed in 40 mL of ethyl acetate. 150 mL of petroleum ether was added, and the mixture was refluxed for 30 min. The mixture was cooled to room temperature, filtered, and dried to obtain 10.8 g of intermediate 3-9, with a yield of 96.4%, which was used directly in the next step.

(7) Preparation of intermediate 3-10

Intermediate 3-9 (12.8 g, 37.0 mmol, 1.0 eq) was added to 100 mL of tetrahydrofuran. The solid was insoluble. 25 mL of water was added to dissolve 4.5 g of sodium hydroxide solution. The mixture was heated to 45-50 °C and reacted overnight. TLC (EA/PE ~ 1:1) showed complete reaction. The reaction was terminated, and the system was concentrated to remove tetrahydrofuran. The system was diluted with 20 mL of water, filtered, and a small amount of tetrahydrofuran was used to wash the solid. The solid was suspended in ethyl acetate and water, and the pH was adjusted to 2-3 with acetic acid. The organic phase was separated, washed with water and brine, dried, concentrated, and slurried with petroleum ether. Filtration and drying yielded 5.7 g of intermediate 3-10, with a yield of 57.2%.

(8) Preparation of intermediate 3-11

Intermediate 3-10 (504 mg, 1.86 mmol) and L-threonine methyl ester hydrochloride (339 mg, 2 mmol) were dissolved in 4 mL of DMF. DIEA (517 mg, 4 mmol) was added, and the solution was stirred for 50 min. After the solution became clear, HOBt (270 mg, 2 mmol) and EDCI (383 mg, 2 mmol) were added, and the reaction was stirred for 15 hours. The post-treatment system was poured into water, extracted twice with 100 mL of ethyl acetate, separated, dried the organic phase, and purified by rotary column chromatography (DCM:MeOH = 100:1-40:1) to obtain 300 mg of product intermediate 3-11.

(9) Preparation of compound 3

The crude intermediate 3-11 (270 mg) was dissolved in 1.5 mL of methanol, and 1 mL of 50% hydroxylamine aqueous solution was added. The reaction was stirred for 24 hours. The post-treatment system was poured into water, filtered, the filter cake was washed with water and dried to obtain 145 mg of compound 3.

_{Molecular formula: C₂₀H₂₂N₂O₆ Molecular weight} : _386.40 Mass spectrometry: (M+H): 387.1

¹ H NMR (DMSO, 401MHz) δ12.84 (s, 1H), 7.90 (d, J=8.4Hz, 3H), 7.59 (d, J=8.3Hz, 3H), 7.20 (dd, J=15.5, 10.8Hz, 2H), 6.90-6.74 (m, 3H), 6.64 (d, J = 15.4Hz, 2H), 6.50 (d, J = 3.2Hz, 2H), 6.35 (d, J = 3.2Hz, 1H), 5.27 (t, J = 5.6Hz, 1H), 4.41 (d, J = 5.4Hz, 3H).

Example 4 4-((5-((dimethylamino)methyl)furan-2-yl)but-1,3-diynyl)-N-((2S,3R)-3- Preparation of hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)benzamide (compound 4)

(1) Preparation of N,N-dimethyl-1-(5-((trimethylsilyl)ethynyl)furan-2-yl)methylamine

5-((trimethylsilyl)ethynyl)furan-2-carboxaldehyde (0.500 g, 2.60 mmol) and 3 mL of dimethylamine in dichloromethane were dissolved in 10 mL of DCM. Sodium triacetoxyborohydride (1.10 g, 5.19 mmol) was added, and the reaction was stirred at room temperature for 1 day. The solution was then evaporated to dryness and used directly in the next step of the reaction.

(2) Preparation of 1-((5-ethynylfuran-2-yl)N,N-dimethylmethylamine

The crude product from the previous step was added to 10 mL of anhydrous methanol, _{and K₂CO₃} (1.435 g, 10.4 mmol) was added. The reaction was stirred at room temperature for 5 hours. The system was filtered, the filtrate was evaporated to dryness, and silica gel column chromatography (PE∶EA＝10∶1--1∶1) was performed to obtain 0.35 g of oil. The two-step yield was 90.3%.

(3) Preparation of methyl (2S,3R)-2-(4-((5-((dimethylamino)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)-3-hydroxybutyrate

CuCl (2 mg, 0.02 mmol) and hydroxylamine hydrochloride (7 mg, 0.1 mmol) were dissolved in 1.5 mL of 23% n-butylamine aqueous solution. Then, 0.5 mL of 23% n-butylamine aqueous solution containing 1-((5-ethynylfuran-2-yl)N,N-dimethylmethylamine (0.20 g, 1.34 mmol) was added. Next, 1.5 mL of methanol and 0.75 mL of tetrahydrofuran solution containing (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate (0.456 g, 1.34 mmol) were added to the above reaction solution. The mixture was stirred for 5 min, then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The product was obtained by silica gel column chromatography (DCM:MeOH = 10:1), yielding 0.26 g of product (yield: 47.5%).

(4) Preparation of 4-((5-((dimethylamino)methyl)furan-2-yl)but-1,3-diynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)benzamide

Methyl (2S,3R)-2-(4-((5-(((dimethylamino)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)-3-hydroxybutyrate (0.23 g, 0.563 mmol) was dissolved in 1 mL of methanol, and 3 mL of 50% hydroxylamine aqueous solution was added. The reaction was stirred at room temperature for 3 hours. The product was prepared directly by liquid phase (methanol:water = 50%), yielding 0.035 g of product, with a yield of 15.2%.

_Molecular formula: _{C₂₂H₂₃N₃O₅ Molecular} weight: _409.2 Mass spectrometry (M+H): 410.0

^1H -NMR(d ₆ -DMSO, 400MHz) δ 8.42 (1H, s), 7.95 (2H, d), 7.70 (2H, d), 7.07 (1H, d), 6.44 (1H, d) ), 4.30-4.21(1H, m), 4.05-3.95(1H, m), 3.45(2H, s), 2.15(6H, s), 1.06(3H, d).

Example 5 N-Methyl-N-(3-methylamino-1-(hydroxyamino)-1,3-dioxopropane-2-yl)-4-((5-(hydroxy) Preparation of (methyl)furan-2-yl)but-1,3-diynyl)benzoamide (compound 5)

(1) Preparation of 5-((trimethylsilyl)ethynyl)furan-2-carboxaldehyde

5-Bromo-2-furfural (25.00 g, 142.85 mmol), trimethylsilylacetylene (20.97 g, 212.9 mmol), triethylamine (28.69 g, 283.5 mmol), Pd( _PPh3 ) _2Cl2 ( _1.0 g, 1.425 mmol), and CuI (0.27 g, 1.415 mmol) were dissolved in 200 mL of THF, purged three times with nitrogen, stirred at room temperature for 3 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 5:1) to give 18.2 g of pale red solid, yield: 66.4%.

(2) Preparation of 5-ethynylfuran-2-carboxaldehyde

5-((trimethylsilyl)ethynyl)furan-2-carboxaldehyde (15.52 g, 80.7 mmol) was dissolved in 300 _mL of methanol, and _K₂CO₃ (33.46 g, 242.1 mmol) was added. The reaction mixture was stirred for 2 hours, evaporated to dryness, and then extracted with 300 mL of ethyl acetate and 300 mL of water. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to give 9.5 g of a yellow solid. Yield: 98%.

(3) Preparation of 5-ethynylfuran-2-yl)methanol

5-Ethynylfuran-2-carboxaldehyde (9.5 g, 79.17 mmol) was dissolved in 200 mL of anhydrous methanol. _NaBH₄ (3.292 g, 87.1 mmol) was slowly added at 0 °C. The reaction was stirred at room temperature for 7 hours. 100 mL of saturated ammonium chloride aqueous solution was added, followed by extraction with 300 mL of ethyl acetate. The mixture was washed with 200 mL of water, dried over anhydrous sodium sulfate, and then evaporated to dryness. Silica gel column chromatography (100% petroleum ether → petroleum ether:ethyl acetate = 2:1) yielded 9.4 g of a colorless oil, with a yield of 97.3%.

(4) Preparation of methyl 4-((trimethylsilyl)ethynyl)benzoate

Methyl 4-iodobenzoate (40 g, 152.6 mmol), trimethylsilylacetylene (14.99 g, 152.6 mmol), triethylamine (35.47 g, 350.5 mmol), Pd( _PPh3 ) _2Cl2 ( _1.071 g, 1.526 mmol), and CuI (0.291 g, 1.526 mmol) were dissolved in 300 mL of THF. The mixture was purged with nitrogen three times, stirred at room temperature for 18 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 1:2) to give 31.5 g of red solid, yield: 88.9%.

(5) Preparation of methyl 4-ethynylbenzoate

Methyl 4-((trimethylsilyl)ethynyl)benzoate (31.5 g, 135.6 mmol) was dissolved in 450 mL of methanol, _{and K₂CO₃} (37.5 g, 271.5 mmol) was added. The reaction mixture was stirred for 2 hours, evaporated to dryness, and then extracted with 450 mL of ethyl acetate and 450 mL of water. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to give 21.75 g of a yellow solid. Yield: 100%.

(6) Preparation of methyl 4-(bromoethynyl)benzoate

Methyl 4-ethynylbenzoate (21.75 g, 135.79 mmol) was dissolved in 500 mL of acetone, silver nitrate (2.176 g, 12.81 mmol) was added, and the mixture was stirred for 40 min. Then, NBS (26.90 g, 151.16 mmol) was added, and the resulting solution was stirred at room temperature for 2 hours. The mixture was filtered, and the filtrate was evaporated to dryness. The solution was then crystallized from isopropanol to give 26.75 g of a white solid. Yield: 82.4%.

(7) Preparation of 4-(bromoethynyl)benzoic acid

Methyl 4-(bromoethynyl)benzoate (26.75 g, 111.89 mmol) was dissolved in 200 mL of methanol and 20 mL of water. NaOH (8.96 g, 224 mmol) was added, and the reaction was stirred for 2 hours. 350 mL of water was added, and the pH was adjusted to 6 with 1 N dilute hydrochloric acid. The mixture was filtered to obtain 18.92 g of white solid. Yield: 75%.

(8) Preparation of 4-(bromoethynyl)benzoyl chloride

4-(bromoethynyl)benzoic acid (8 g, 35.56 mmol) was dissolved in 200 mL of dichloromethane, and 30 mL of thionyl chloride was added. The mixture was stirred for 5 hours and then evaporated to dryness to obtain 8.658 g of a yellow solid, with a yield of 100%.

(9) Preparation of diethyl 2-((benzyl)methylamino)-malonic acid

Diethyl 2-bromomalonate (80 g, 33.46 mmol) was added to 200 mL of acetonitrile, followed by methylbenzylamine (81 g, 66.92 mmol). The mixture was stirred at room temperature for 5 hours until the reaction was complete. The white solid was removed by filtration, and most of the filtrate was discarded by rotary evaporation. 100 mL of toluene was added to the system, and the white solid was removed by filtration. The filtrate was collected and evaporated to dryness for later use, yielding 67 g of a colorless oily substance, with a yield of 71.7%.

(10) Preparation of ethyl 2-((benzyl)methylamino)-3-methylaminomalonic acid

Diethyl 2-((benzyl)methylamino)-malonate (50 g, 89.5 mmol) was dissolved in 200 mL of methanol, and 30 mL of methylamino alcohol solution was added. The mixture was stirred overnight at room temperature. After the reaction was complete, it was added to 1000 mL of water, extracted three times with ethyl acetate, evaporated to dryness, and column chromatography was used to obtain 12 g of the intermediate ethyl 2-((benzyl)methylamino)-3-methylaminomalonate, with a yield of 25.4% (PE→PE∶EA＝5∶1).

(11) Preparation of ethyl 2-((tert-Butoxycarbonyl)methylamino)-3-methylaminomalonic acid

Ethyl 2-((benzyl)methylamino)-3-methylaminomalonate (12 g, 45.4 mmol) was dissolved in 200 mL of methanol, and 1.2 g of palladium hydroxide on carbon and di-tert-butyl dicarbonate (9.9 g, 45.4 mmol) were added. The reaction was carried out overnight at room temperature under hydrogen protection. After the reaction was completed, the mixture was filtered through diatomaceous earth, and the filtrate was collected. The solution was then evaporated to dryness and column chromatography to give 6.4 g of intermediate, with a yield of 51.4% (PE→PE∶EA＝20∶1).

(12) Preparation of ethyl 2-methylamino-3-methylaminomalonic acid

Ethyl 2-((tert-Butoxycarbonyl)methylamino)-3-methylaminomalonate (6.4 g, 23.33 mmol) was dissolved in 100 mL of dichloromethane, and 30 mL of trifluoroacetic acid was added. The mixture was stirred overnight at room temperature until the reaction was complete. The solution was then evaporated to dryness to obtain a brown oily substance, 4.064 g (crude product), with a yield of 100%.

(13) Preparation of ethyl 2-(N-(4-bromoethynylbenzoyl)-N-methylamino)-3-methylaminomalonic acid

Ethyl 2-methylamino-3-methylaminomalonate (4.064 g, 23.3 mmol) was dissolved in 100 mL of THF, and triethylamine (7.06 g, 69.9 mmol) and 4-(bromoethynyl)benzoyl chloride (5.674 g, 23.3 mmol) were added. The mixture was stirred overnight at room temperature. After the reaction was completed, the product was evaporated to dryness and then subjected to column chromatography to obtain 5.2 g of a brown oily substance, with a yield of 58.6% (DCM→DCM∶MeOH＝30∶1).

(14) Preparation of ethyl 2-((4-(5-(hydroxymethyl)-tetrahydrofuran-2-yl)but-1,3-diynyl)-N-benzoylamino-N-methylamino)-3-methylaminomalonate

CuCl (3 mg, 0.03 mmol) and hydroxylamine hydrochloride (7 mg, 0.1 mmol) were dissolved in 12 mL of 23% n-butylamine aqueous solution. Then, 5 mL of methanol and tetrahydrofuran solution (V:V, 1:1) containing (5-ethynylfuran-2-yl)methanol (0.220 g, 1.80 mmol) and 5 mL of methanol and tetrahydrofuran solution (V:V, 1:1) containing ethyl 2-(N-(4-bromoethynylbenzoyl)-N-methylamino)-3-methylaminomalonate (0.629 g, 1.65 mmol) were added to the above reaction solution. The mixture was stirred for 2 min, then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. Silica gel column chromatography (100% petroleum ether → petroleum ether:ethyl acetate = 2:1) yielded 0.49 g of a brown oily substance, yield: 70.3%.

(15) Preparation of N-methyl-N-(3-methylamino-1-(hydroxyamino)-1,3-dioxopropane-2-yl)-4-((5-(hydroxymethyl)furan-2-yl)but-1,3-diynyl)benzoamide

Ethyl 2-((4-(5-(hydroxymethyl)-tetrahydrofuran-2-yl)but-1,3-diynyl)-N-benzoamide-N-methylamino)-3-methylaminomalonate (0.49 g, 1.16 mmol) was dissolved in 10 mL of methanol, and 2 mL of 50% hydroxylamine aqueous solution was added. The reaction was stirred for 24 hours, and the product was post-processed to obtain compound 37, totaling 132 mg, with a yield of 27.8%.

_{Molecular formula: C₂₁H₁₉N₃O₆ Molecular weight} : _409.1 Mass spectrometry: (2M+H): 819.3

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.79 (1H, s), 9.05 (1H, s), 8.19 (1H, m), 7.71 (2H, m), 7.41 (2H, m), 7.07 (1 H, m), 6.45 (1H, m), 5.43 (1H, m), 5.35 (1H, s), 4.41 (2H, m), 2.66 (3H, m), 2.49 (3H, m).

Example 6: N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl)) Preparation of furan-2-yl)but-1,3-diynyl)benzoamide (compound 6)

Following the preparation of compound 3 above, compound 6 was synthesized using methyl (2S,3R)-2-(4-(bromoethynyl)benzoamide)-3-hydroxybutyrate.

_{Molecular formula: C₂₀H₁₈N₂O₆ Molecular weight} : _382.1 Mass spectrometry (M+H): 383.1

¹ H-NMR(d ₆ -DMSO+CF ₃ COOD, 400MHz) δ 8.19 (1H, d), 7.93 (2H, d), 7.70 (2H, d), 7.03 (1H, d), 6.42 (1H, d) ), 4.40 (2H, s), 4.29-4.21 (1H, m), 4.06-3.98 (1H, m), 3.58 (1H, t), 1.07 (3H, d).

Following the preparation of compound 3 above, compound 7 was synthesized using methyl 2-(4-(bromoethynyl)benzoamide)-3-hydroxybutyrate.

Referring to the preparation of compound 3 above, N-((2R,3S)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl)furan-2-yl)but-1,3-diynyl)benzoamide (compound 8) was synthesized using methyl (2R,3S)-2-(4-(bromoethynyl)benzoamide)-3-hydroxybutyrate.

Referring to the preparation of compound 3 above, N-((2R,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl)furan-2-yl)but-1,3-diynyl)benzoamide (compound 9) was synthesized using methyl (2R,3R)-2-(4-(bromoethynyl)benzoamide)-3-hydroxybutyrate.

Referring to the preparation of compound 3 above, N-((2S,3S)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl)furan-2-yl)but-1,3-diynyl)benzoamide (compound 10) was synthesized using methyl (2S,3S)-2-(4-(bromoethynyl)benzoamide)-3-hydroxybutyrate.

Compounds 6 and 8-10 are stereoisomers of compound 7, and their ^1H -NMR data are basically the same, with no obvious differences.

High-performance liquid chromatography (HPLC)

HPLC conditions:

Instrument: High-performance liquid chromatograph; Column: Agilent ZORBAX SB-C18 (4.6 mm × 100 mm, 3.5 μm); Wavelength = 286 nm; Column temperature = 30 °C; Flow rate = 1.0 mL/min; Injection volume = 10 μL; Gradient elution;

Mobile phase A: Accurately measure 0.8 mL of trifluoroacetic acid, add it to 1000 mL of water and mix well.

Mobile phase B: Accurately measure 0.5 mL of trifluoroacetic acid and mix it with 1000 mL of methanol.

Test solution: Weigh appropriate amounts of compounds 6, 9 and 10 accurately, dissolve them in methanol to prepare a solution containing approximately 0.5 mg per mL, which is used as the test solution.

Experimental results: The retention time of compound 6 was 26.601 min;

Compound 9 had a retention time of 28.799 min;

The retention time of compound 10 was 29.051 min.

Example 7 4-((5-((cyclopropylamino)methyl)furan-2-yl)but-1,3-diynyl)-N-((2S,3R)- Preparation of 3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)benzamide (compound 11)

Compound 11 was synthesized following the preparation method of compound 3 described above.

_{Molecular formula: C₂₃H₂₃N₃O₅ Molecular weight} : _421.2 Mass spectrometry (M+H): 422.0

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.70 (1H, s), 9.19 (1H, br), 8.89 (1H, s), 8.20 (1H, d), 7.94 (2H, d), 7.72 (2H, d), 7.17 (1H, d), 6.74 (1H, d), 4.94 (1H , m), 4.36 (2H, s), 4.26-4.21 (1H, m), 4.04-3.98 (1H, m), 2.75-2.60 (1H, m), 1.07 (3H, d), 0.87-0.79 (2H, m), 0.76-0.70 (2H, m).

Example 8 4-((5-((3-aminopropylamino)methyl)furan-2-yl)but-1,3-diynyl)-N-((2S, Preparation of 3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)benzamide (compound 12)

(1) Preparation of tert-butyl 3-((5-((trimethylsilyl)ethynyl)furan-2-yl)methyleneamino)propylcarbamate

5-((trimethylsilyl)ethynyl)furan-2-carboxaldehyde (0.500 g, 2.60 mmol) was dissolved in 10 mL of methanol, 3-aminopropylcarbamate tert-butyl ester (0.905 g, 5.20 mmol) was added, 1 drop of acetic acid was added, and the mixture was stirred overnight at room temperature and used directly in the next step of the reaction.

(2) Preparation of tert-butyl 3-((5-((trimethylsilyl)ethynyl)furan-2-yl)methylamino)propylcarbamate

Sodium borohydride (0.197 g, 5.20 mmol) was added to the reaction system of the previous step, and the mixture was stirred at room temperature for 1 hour. LC-MS showed that the reaction was complete and no further processing was required. The mixture was then used directly for the next reaction.

(3) Preparation of tert-butyl 3-((5-ethynylfuran-2-yl)methylamino)propylcarbamate

Potassium carbonate (0.717 g, 5.20 mmol) was added to the reaction system of the previous step, and the reaction was carried out at room temperature for 4 hours. After the reaction was completed, 100 mL of water was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to column chromatography (PE∶EA＝5∶1→DCM∶MeOH＝10∶1) to give 0.35 g of oil. The three-step yield was 48.5%.

(4) Preparation of methyl (2S,3R)-2-(4-((5-((3-(tert-butoxycarbonylamino)propylamino)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)-3-hydroxybutyrate

CuCl (3 mg, 0.03 mmol) and hydroxylamine hydrochloride (7 mg, 0.1 mmol) were dissolved in 2 mL of 23% n-butylamine aqueous solution. Then, 5 mL of methanol and tetrahydrofuran solution (V:V, 1:1) containing 3-((5-ethynylfuran-2-yl)methylamino)propylcarbamate tert-butyl ester (0.33 g, 1.19 mmol) were added sequentially, followed by (2S,3R)-2-(4-(bromoethynyl)benzoylamino) A solution of methyl 3-hydroxybutyrate (0.404 g, 1.19 mmol) in 5 mL of methanol and tetrahydrofuran (V:V, 1:1) was added to the above reaction solution, stirred for 2 min, and then 20 mL of dichloromethane and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The solution was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1 → dichloromethane:methanol = 10:1) to give 0.38 g of a brownish-red solid, yield: 59.4%.

(5) Preparation of 4-((5-((3-aminopropylamino)methyl)furan-2-yl)but-1,3-diynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)benzamide

Methyl (2S,3R)-2-(4-((5-((tert-butoxycarbonyl(2-(2-hydroxyethoxy)ethyl)amino)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)-3-hydroxybutyrate (0.38 g, 0.707 mmol) was dissolved in 10 mL of dichloromethane, and 2 mL of TFA was added. The reaction mixture was stirred for 30 min, evaporated to dryness, and the resulting oily residue was added to 1 mL of methanol. 4 mL of 50% hydroxylamine aqueous solution was added, and the reaction mixture was stirred for 3 hours. Direct liquid-phase purification yielded 79 mg of compound 7, with a yield of 25.5%.

_Molecular formula: _{C₂₃H₂₆N₄O₅ Molecular} weight: _438.2 Mass spectrometry (M+H): 439.0

¹ H-NMR (DMSO-d ₆ , 400MHz) δ8.24 (1H, d), 7.94 (2H, d), 7.71 (2H, d), 7.05 (1H, d), 6.39 (1H, d), 4.25 (1H, dd), 4.0 1(1H, quintet), 3.66(2H, s), 3.43(4H, m), 2.68-2.52(2H, m), 1.54-1.40(2H, m), 1.07(3H, d).

Example 9 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((4-methyl) Preparation of piperazine-1-yl)methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 13)

(1) Preparation of 1-methyl-4-((5-((trimethylsilyl)ethynyl)furan-2-yl)methyl)piperazine

In a dry reaction flask, 5-((trimethylsilyl)ethynyl)furan-2-carboxaldehyde (308 mg, 1.6 mmol) and 1-methylpiperazine (192 mg, 1.92 mmol) were dissolved in 20 mL of dichloromethane, and sodium triacetoxyborohydride (509 mg, 2.4 mmol) was added. The mixture was stirred at room temperature for 20 h, and the dichloromethane was removed by concentration to obtain an oily substance that could be used directly in the next reaction step.

(2) Preparation of 1-((5-ethynylfuran-2-yl)methyl)-4-methylpiperazine

In a dry reaction flask, the crude product obtained in the previous step was dissolved in 20 mL of methanol, potassium carbonate (442 mg, 3.2 mmol) and lithium hydroxide (200 mg) were added. After the addition was complete, the mixture was stirred at room temperature for 1 h. The organic phase was then concentrated under reduced pressure and column chromatography (100% EA) was performed to obtain 320 mg of pale yellow solid, with a yield of 97.9%.

Preparation of (3) (2S,3R)-3-hydroxy-2-(4-((5-(((4-methylpiperazin-1-yl)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)methyl butyrate

In a dry reaction flask, CuCl (28 mg, 0.284 mmol) and hydroxylamine hydrochloride (296 mg, 4.26 mmol) were dissolved in 3 mL of 23% n-butylamine aqueous solution. Then, 1 mL of 23% n-butylamine aqueous solution containing 1-((5-ethynylfuran-2-yl)methyl)-4-methylpiperazine (290 mg, 1.42 mmol) was added. Next, 6 mL of methanol and 3 mL of tetrahydrofuran solution containing (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate (340 mg, 1 mmol) were added to the above reaction solution. The mixture was stirred for 5 min, then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to silica gel column chromatography (DCM:MeOH = 20:1) to obtain 360 mg of the product, yield: 54.7%.

(4) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-((4-methylpiperazin-1-yl)methyl)furan-2-yl)but-1,3-diynyl)benzamide

Methyl (2S,3R)-3-hydroxy-2-(4-((5-(((4-methylpiperazin-1-yl)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)butyrate (360 mg, 0.78 mmol) was dissolved in 5 mL of methanol, and 3 mL of 50% hydroxylamine aqueous solution was added. The reaction was carried out at room temperature with stirring for 8 hours, followed by 12 hours in a refrigerator. The product was then purified by liquid chromatography (methanol:water = 55%) to yield 40 mg of the product, yield: 11.0%.

_Molecular formula: _{C₂₅H₂₈N₄O₅ Molecular} weight: _464.2 Mass spectrometry (M+H): 465.2

^1H -NMR(d ₆ -DMSO, 400MHz) δ 10.70 (1H, s), 8.88 (1H, s), 8.23 (1H, d), 7.96 (2H, d), 7.74 (2H, d), 7.09 (1H, d), 6.46 (1 H, d), 4.90 (1H, d), 4.26 (1H, dd), 4.03 (1H, q), 3.53 (2H, s), 2.45-2.23 (8H, m), 2.14 (3H, s), 1.09 (3H, d).

Example 10 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-phenylfuran) Preparation of (2-yl)but-1,3-diynyl)benzamide (compound 14)

Referring to Example 9, compound 14 was synthesized.

Molecular formula: _{C₂₅H₂₀N₂O₅ Molecular} weight: _428.1 Mass spectrometry: (M+H): 429.1 _, 396.1

¹ H-NMR (DMSO-d ₆ , 600MHz) δ10.67 (1H, s), 8.84 (1H, s), 8.20 (1H, d), 7.95 (2H, d), 7.77 (2H, d), 7.74 (2H, d), 7.47 (2H , t), 7.37 (1H, t), 7.26 (1H, d), 7.14 (1H, d), 4.90 (1H, s), 4.25 (1H, dd), 4.02 (1H, dt), 1.08 (3H, d).

Example 11 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((methylamino) Preparation of (methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 15)

Referring to Example 3, compound 15 was synthesized.

Molecular formula: C21H21N3O5 Molecular weight: 395.1 Mass spectrometry (M+H): 396.2

1H-NMR (d ₆ -DMSO, 400MHz) δ 10.75 (1H, br s), 8.87 (1H, s), 8.23 (1H, d), 7.96 (2H, d), 7.74 (2H, d), 7.08 (1H, d), 6.42 (1H, d) , 4.90 (1H, d), 4.26 (1H, dd), 4.03 (1H, q), 3.65 (2H, s), 2.26 (3H, s), 1.09 (3H, d).

Example 12 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl) Preparation of (-4-methylfuran-2-yl)but-1,3-diynyl)benzamide (compound 16)

Compound 16 was synthesized following the preparation method of compound 3 described above.

_Molecular formula: _{C₂₁H₂₀N₂O₆ Molecular} weight: _396.1 Mass spectrometry (M+H): 397.1

¹ H-NMR (DMSO-d ₆ , 600MHz) δ 7.92 (2H, d), 7.67 (2H, d), 6.88 (1H, s), 4.33 (2H, s), 4.23 (1H, dd), 4.00 (1H, dt), 1.95 (3H, s), 1.06 (3H, d).

Example 13 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl) Preparation of (-3-phenylfuran-2-yl)but-1,3-diynyl)benzamide (compound 17)

Compound 17 was synthesized following the preparation method of compound 3 described above.

_{Molecular formula: C₂₆H₂₂N₂O₆ Molecular weight} : _458.1 Mass spectrometry (M+H): 459.1

¹ H-NMR (DMSO-d ₆ , 600MHz) δ10.67 (1H, s), 8.85 (1H, s), 8.20 (1H, d), 7.93 (2H, d), 7.77 (2H, d), 7.74 (2H, d), 7.48 (2H, t), 7 .38(1H,t), 6.90(1H,s), 5.51(1H,t), 4.88(1H,d), 4.43(2H,d), 4.23(1H,dd), 4.00(1H,dt), 1.06(3H,d).

Example 14 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((2-(2- Preparation of hydroxyethoxy)ethylamino)methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 18)

Referring to Example 8, compound 18 was synthesized.

_{Molecular formula: C24H27N3O7 Molecular} weight: _469.2 Mass spectrometry (M+H): ₄₇₀

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.66 (1H, s), 8.84 (1H, s), 8.19 (1H, d), 7.93 (2H, d), 7.71 (2H, d), 7.05 (1H, d), 6.40 (1H, d), 4.88 (1H, d), 4.64- 4.52(1H,m), 4.24(1H,dd), 4.06-3.95(1H,m), 3.71(2H,s), 3.50-3.41(4H,m), 3.40-3.35(3H,m), 2.64(2H,t), 1.07(3H,d).

Example 15 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((2-(2- Preparation of hydroxyethoxy)ethylamino)methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 19)

Referring to Example 8, compound 19 was synthesized.

_Molecular formula: _{C₂₄H₂₇N₃O₆ Molecular} weight: _453.2 Mass spectrometry (M+H): 454.0

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.69 (1H, s), 8.85 (1H, s), 8.22 (1H, d), 7.95 (2H, d), 7.71 (2H, d), 7.06 (1H, d), 6.42 (1H, d), 4.90 (1H, d), 4.28- 4.20(1H,m), 4.08-3.97(1H,m), 3.72(2H,s), 3.49(1H,s), 3.36(2H,m), 2.05-1.92(2H,m), 1.46-1.39(4H,m), 1.07(3H,d).

Example 16 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-(furan-2-yl) Preparation of but-1,3-diynyl)benzoamide (compound 20)

Referring to Example 3, compound 20 was synthesized.

Molecular formula: C19H16N2O5 Molecular weight: 352.1 Mass spectrometry (M+H): 353.1

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.7 (1H, S), 8.89 (1H, S), 8.30 (1H, S), 8.23 (1H, m), 7.96 (2H, m), 7.82 (1 H, s), 7.72 (2H, m), 6.76 (1H, s), 4.92 (1H, s), 4.27 (1H, m), 4.04 (1H, s), 1.1 (3H, m).

Example 17 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((2-(2- Preparation of hydroxyethoxy)ethoxy)methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 21)

Referring to Example 8, compound 21 was synthesized.

_{Molecular formula: C₂₄H₂₆N₂O₈ Molecular weight} : _470.2 Mass spectrometry (M+H): 471.2

¹ H-NMR (DMSO-d ₆ , 400MHz) 10.72 (1H, s), 8.90 (1H, s), 8.21 (1H, d), 7.95 (2H, d), 7.74 (2H, d), 7.10 (1H, d), 6.59 (1H, d) ), 4.94(1H, d), 4.64(1H, t), 4.47(2H, s), 4.25(1H, dd), 4.03(1H, q), 3.68-3.52(8H, m), 1.09(3H, d).

Example 18 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((3-morpholine) Preparation of propylamino)methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 22)

(1) Preparation of 3-morpholino-N-((5-((trimethylsilyl)ethynyl)furan-2-yl)methyl)prop-1-amine

192 mg (1 mmol) of 5-((trimethylsilyl)ethynyl)furan-2-carboxaldehyde and 144 mg (1 mmol) of 3-morpholinopropyl-1-amine were added to 10 mL of methanol, followed by 3 drops of glacial acetic acid. The reaction was carried out at 40 °C for 18 h. Subsequently, 57 mg (1.5 mmol) of sodium borohydride was added, and the reaction was carried out for 4 h. The same reaction was scaled up 2.6 times, the results were combined, and the mixture was directly evaporated to dryness for use in the next reaction.

(2) Preparation of tert-butyl 3-morpholinopropyl ((5-((trimethylsilyl)ethynyl)furan-2-yl)methyl)carbamate

The product from the previous step was dissolved in 30 mL of dichloromethane, and 505 mg (5 mmol) of triethylamine and 872 mg (4 mmol) of di-tert-butyl dicarbonate were added. The mixture was reacted at room temperature for 24 h. The solution was evaporated to dryness and subjected to column chromatography (PE:EA = 3:1) to give 1.06 g of a yellow solid.

(3) Preparation of (5-ethynylfuran-2-yl)methyl(3-morpholinopropyl)carbamate tert-butyl ester

1.06 g (2.52 mmol) of 3-morpholinopropyl ((5-((trimethylsilyl)ethynyl)furan-2-yl)methyl)carbamate tert-butyl ester was dissolved in 30 mL of methanol, and 1.04 g (7.52 mmol) of anhydrous potassium carbonate was added. The mixture was stirred at room temperature for 8 h. After filtration, the solvent was evaporated to dryness to give 1.1 g of crude product as a yellow solid.

(4) Preparation of (2S,3R)-2-(4-((5-((tert-butoxycarbonyl(3-morpholinopropyl)amino)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)-3-hydroxybutyrate methyl ester

Under ice bath conditions, 2 mg of cuprous chloride and 4 mg of hydroxylamine hydrochloride were added separately to 2 mL of 23% butylamine aqueous solution. 209 mg (0.6 mmol) of (5-ethynylfuran-2-yl)methyl(3-morpholinopropyl)carbamate tert-butyl ester dissolved in 2 mL of 23% butylamine aqueous solution was added dropwise to the system. Then, 170 mg (0.5 mmol) of (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate methyl ester was dissolved in a mixed solvent of 1 mL butylamine aqueous solution, 1 mL methanol, and 1 mL tetrahydrofuran, and added dropwise to the system. The reaction was allowed to proceed for 1 min. Water was added, and the mixture was extracted with ethyl acetate, evaporated to dryness, and subjected to column chromatography (DCM:MeOH = 20:1) to give 144 mg of a white solid, yield 47.4%.

(5) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-(((3-morpholinopropylamino)methyl)furan-2-yl)but-1,3-diynyl)benzamide

144 mg (0.237 mmol) of methyl (2S,3R)-2-(4-((5-((tert-butoxycarbonyl(3-morpholinopropyl)amino)methyl)furan-2-yl)but-1,3-diynyl)benzoylamino)-3-hydroxybutyrate was dissolved in 4 mL of dichloromethane, and 1 mL of trifluoroacetic acid was added. The mixture was reacted at room temperature for 4 h. The solution was then evaporated to dryness and used directly in the next reaction.

The product from the previous step was dissolved in 8 mL of anhydrous methanol, and 3 mL of 50% hydroxylamine aqueous solution was added. The mixture was reacted at room temperature for 3 h, and then directly purified by liquid phase to obtain 20 mg of compound 16, with a yield of 16.6%.

Molecular formula: C ₂₇ H ₃₂ N ₄ O ₆ Molecular weight: 508.2 Mass spectrometry (M+H): 509.1

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.65 (1H, s), 8.84 (1H, s), 8.19 (1H, d), 7.93 (2H, d), 7.71 (2H, d), 7.05 (1H, d), 6.39 (1H, d), 4.87 (1H, d), 4.24 ( 1H, dd), 4.05-3.95 (1H, m), 3.67 (2H, s), 3.54 (4H, t), 3.30-3.24 (2H, m), 2.35-2.20 (6H, m), 1.53 (2H, quintet, 1.07 (3H, d).

Example 19 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(morpholinomethyl) Preparation of (2-( ...

Compound 23 was synthesized according to Example 9.

_Molecular formula: _{C₂₄H₂₅N₃O₆ Molecular} weight: _451.2 Mass spectrometry (M+H): 452.0

^1H -NMR(d ₆ -DMSO, 400MHz) δ 10.67 (1H, s), 8.85 (1H, d), 8.20 (1H, d), 7.94 (2H, d), 7.72 (2H, d), 7.07 (1H, d), 6.47 (1 H, d), 4.88 (1H, d), 4.24 (1H, dd), 4.08-3.95 (1H, m), 3.55 (4H, t), 3.53 (2H, s), 2.37 (4H, t), 1.07 (3H, d).

Example 20 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(piperazine-1- Preparation of (methyl)furan-2-yl)but-1,3-diynyl)benzamide (compound 24)

Compound 24 was synthesized according to Example 9.

_{Molecular formula: C₂₄H₂₆N₄O₅ Molecular weight} : _450.2 Mass spectrometry (M+H): 451.0

¹ H-NMR (DMSO-d ₆ , 400MHz) δ10.68 (1H, s), 8.85 (1H, s), 8.20 (1H, d), 7.94 (2H, d), 7.72 (2H, d), 7.06 (1H, d), 6.44 (1H, d), 4.88(1H,d), 4.24(1H,dd), 4.05-3.95(1H,m), 3.49(2H,s), 2.69(4H,t), 2.04-1.92(4H,m), 1.07(3H,d).

Example 21 4-(3-(furan-2-yl)acrylamido)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1- Preparation of oxobutane-2-yl)-benzamide (compound 30)

(1) Preparation of intermediate 21-1

2-Furanacrylic acid (2.762 g, 20 mmol) was dissolved in 30 mL of dichloromethane, and thionyl chloride (2.379 g, 20 mmol) was added. The mixture was stirred at 25 °C for one hour, and then p-aminobenzoic acid (2.743 g, 20 mmol) was added. The mixture was heated to 35 °C and reacted for 72 hours. The post-treatment system was evaporated to dryness and purified by column chromatography (PE:EA = 2:1) to give 514 mg of intermediate 21-1.

(2) Preparation of intermediate 21-2

Intermediate 21-1 (514 mg, 2 mmol) and L-threonine methyl ester hydrochloride (339 mg, 2 mmol) were dissolved in 4 mL of DMF. DIEA (517 g, 4 mmol) was added, and the solution was stirred for 50 min. After the solution became clear, HOBt (270 mg, 2 mmol) and EDCI (383 mg, 2 mmol) were added, and the reaction was stirred for 15 hours. The post-treatment system was poured into water, extracted with ethyl ester, separated, and the organic phase was dried under evaporation to obtain 800 mg of crude product. The crude product was purified by column chromatography (PE:EA = 1:2) to obtain 200 mg of intermediate 21-2.

(3) Preparation of 4-(3-(furan-2-yl)acrylamido)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-benzamide (compound 30)

Intermediate 21-2 (150 mg, 0.403 mmol) was dissolved in 4 mL of methanol, and 2 mL of 50% hydroxylamine aqueous solution was added. The reaction was stirred for 40 hours. The post-treatment system was poured into water, filtered, the filter cake was washed with water and dried to obtain 74 mg of compound 30.

_Molecular formula: _{C18H19N3O6 Molecular} weight: _373.36 Mass spectrometry (M+H): 374.0

^1H -NMR(d ₆ -DMSO, 800MHz) δ 10.62 (1H, s), 10.1 (1H, s), 8.81 (1H, s), 7.83 (4H, d), 7.74 (2H, d), 7.4 0(1H,d), 6.86(1H,s), 6.62(2H,d), 4.87(1H,d), 4.23(1H,d), 4.01(1H,d), 1.10(3H,d).

Example 22 4-(2-(furan-2-methylene)hydrazine carbonyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1- Preparation of oxobutane-2-yl)-benzamide (compound 26)

Referring to Example 21, compound 26 was synthesized.

_Molecular formula: _{C17H18N4O6 Molecular} weight: _374.1 Mass spectrometry (M+H): 375.0

¹ H-NMR (DMSO-d ₆ , 800MHz): δ11.89 (1H, s), 8.43 (1H, s), 8.34 (1H, s), 7.99 (4H, s), 7.85 (1H, s), 6.94 ( 1H, s), 6.93 (1H, s), 4.99 (1H, s), 4.50 (1H, s), 4.17 (1H, s), 3.64 (1H, s), 1.13 (4H, d)

Example 23 5-{4-[N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)aminomethyl] Preparation of [acyl]-phenyl]-but-1,3-diynyl]-furan-2-carboxylic amide (compound 27)

Compound 27 was synthesized according to Example 4 above.

_Molecular formula: _{C₂₀H₁₇N₃O₆ Molecular} weight: _395.37 Mass spectrometry (M+H): 396.0

^1H -NMR(d ₆ -DMSO, 800MHz) δ10.69 (1H, s), 8.87 (1H, s), 8.25 (1H, d), 8.0 (1H, s), 7.97 (2H, d), 7.77 (2H, d), 7.76 (1H, s), 7.22 (1H, q), 4.89 (1H, d), 4.25 (1H, t), 4.03 (1H, q), 1.09 (3H, t).

Example 24 5-{4-[N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)aminomethyl] Preparation of [acyl]-phenyl]-but-1,3-diynyl]-furan-2-carboxylic acid dicarboxamide (compound 29)

Referring to Example 4 above, compound 29 was synthesized.

_{Molecular formula: C₂₂H₂₁N₃O₆ Molecular weight} : _423.42 Mass spectrometry (M+H): 424.1

^1H -NMR(d ₆ -DMSO, 800MHz) δ 10.69 (1H, s), 8.87 (1H, s), 8.25 (1H, d), 7.96 (2H, d), 7.76 (2H, d), 7.25 (1H, d) ), 7.12(1H,d), 4.89(1H,d), 4.25(1H,t), 4.03(1H,q), 3.19(3H,s), 2.98(3H,s), 1.09(3H,d).

Example 25 4-((1E,3E)-4-(furan-3-yl)but-1,3-dienyl)-N-((2S,3R)-3-hydroxy-1- Preparation of (hydroxyamino)-1-oxobutane-2-yl)benzamide (compound 31)

Compound 31 was synthesized according to Example 1 above.

_Molecular formula: _{C19H20N2O5 Molecular} weight: _356.37 Mass spectrometry (M+H): 357.0

^1H -NMR(d ₆ -DMSO, 800MHz) δ 10.66 (1H, s), 8.85 (1H, d), 7.98 (1H, d), 7.90-7.85 (3H, m), 7.68 (1H, d), 7.58 (2H, d), 7.22- 7.14(1H,m), 6.84-6.78(2H,m), 6.71-6.66(2H,m), 4.89(1H,d), 4.27(1H,t), 4.06-4.0(1H,m), 1.17(3H,d).

Example 26 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-6-((1E,3E)-4- Preparation of ((5-(hydroxymethyl)-furan-2-yl)but-1,3-dienyl)nicotinamide (compound 32)

Referring to Example 1 above, compound 32 was synthesized _. Molecular formula: _{C₂₀H₂₂ON₂O₆ Molecular} weight: _386.40 Mass spectrometry (M+H): 387.1

^1H -NMR(d ₆ -DMSO, 800MHz) δ10.65 (1H, s), 8.99 (1H, d), 8.88 (1H, s), 8.30-8.20 (2H, m), 7.58-7.25 (1H, m), 6.86-6.79 (3H, m), 6.64 (1H, d), 6.49 (1H, d), 6.38 (1H, d), 5.31-5.28 (1H, m), 4.89 (1H, s), 4.3 (2H, s), 4.31-4.26 (1H, m), 4.06-4.03 (1H, m), 1.10 (3H, d).

Example 27 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl) Preparation of (-1-methyl-1H-pyrrole-2-yl)but-1,3-diynyl)benzoamide (compound 33)

(1) Preparation of methyl 4-((trimethylsilyl)ethynyl)benzoate

Methyl 4-iodobenzoate (8.00 g, 30.5 mmol), trimethylsilylacetylene (2.995 g, 30.5 mmol), triethylamine (7.09 g, 70.1 mmol), Pd( _PPh3 ) _2Cl2 ( _0.214 g, 0.305 mmol), and CuI (0.057 g, 0.299 mmol) were dissolved in 60 mL of THF, purged three times with nitrogen, stirred at room temperature for 18 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 1:2) to give 6.3 g of red solid, yield: 88.9%.

(2) Preparation of methyl 4-ethynylbenzoate

Methyl 4-((trimethylsilyl)ethynyl)benzoate (2.10 g, 9.04 mmol) was dissolved in 30 mL of methanol, _{and K₂CO₃} (2.50 g, 18.1 mmol) was added. The reaction mixture was stirred for 2 hours, evaporated to dryness, and then extracted with 30 mL of ethyl acetate and 30 mL of water. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to give 1.45 g of a yellow solid. Yield: 100%.

(3) Preparation of methyl 4-(bromoethynyl)benzoate

Methyl 4-ethynylbenzoate (1.00 g, 6.24 mmol) was dissolved in 50 mL of acetone, silver nitrate (0.10 g, 0.58 g) was added, and the mixture was stirred for 40 min. Then, NBS (1.237 g, 6.95 mmol) was added, and the resulting solution was stirred at room temperature for 2 hours. The mixture was filtered, and the filtrate was evaporated to dryness. The solution was then crystallized from isopropanol to give 1.23 g of white solid. Yield: 82.4%.

(4) Preparation of 4-(bromoethynyl)benzoic acid

Methyl 4-(bromoethynyl)benzoate (1.23 g, 5.14 mmol) was dissolved in 9 mL of methanol and 1 mL of water. NaOH (0.413 g, 10.3 mmol) was added, and the reaction was stirred for 2 hours. 15 mL of water was added, and the pH was adjusted to 6 with 1 N dilute hydrochloric acid. The mixture was filtered to obtain 0.87 g of white solid. Yield: 75.3%.

(5) Preparation of methyl (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate

4-(bromoethynyl)benzoic acid (0.87 g, 3.87 mmol) and L-threonine methyl ester hydrochloride (0.693 g, 4.09 mmol) were dissolved in 10 mL of DMF. DIEA (0.957 g, 7.41 mmol) was added, and the solution was stirred for 30 min. After the solution became clear, HOBt (0.552 g, 4.09 mmol) and EDCI (0.784 g, 4.09 mmol) were added, and the reaction was stirred for 15 h. 30 mL of ethyl acetate and 30 mL of water were added, and the mixture was extracted. The resulting organic phase was dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 1:1) to give 1.21 g of white solid, yield 92%.

(6) Preparation of methyl 5-bromo-1-methyl-1H-pyrrole-2-carboxylate

1-Methylpyrrole-2-carboxylic acid methyl ester (3.0 g, 21.6 mmol) and NBS (3.84 g, 21.6 mmol) were dissolved in 40 mL of dichloromethane and reacted in the dark for 12 hours. The product was evaporated to dryness and passed through a silica gel column (petroleum ether) to give 2.21 g of product, with a yield of 46.8%.

(7) Preparation of methyl 1-methyl-5-((trimethylsilyl)ethynyl)-1H-pyrrole-2-carboxylate

Methyl 5-bromo-1-methyl-1H-pyrrole-2-carboxylate (2.00 g, 9.17 mmol), trimethylsilylacetylene (1.08 g, 11.0 mmol), triethylamine (1.86 g, 18.4 mmol), Pd( _PPh3 ) _2Cl2 ( ₄₅ mg, 0.064 mmol), and CuI (4 mg, 0.018 mmol) were dissolved in 20 mL of THF, purged three times with nitrogen, heated to 90 °C and stirred for 18 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 5:1) to give 1.92 g of pale red solid, yield: 89%.

(8) Preparation of methyl 5-ethynyl-1-methyl-1H-pyrrole-2-carboxylate

1-Methyl-5-((trimethylsilyl)ethynyl)-1H-pyrrole-2-carboxylate (1.90 g, 8.07 mmol) was dissolved in 30 mL of methanol, and _K₂CO₃ (2.23 g, 16.2 mmol) was added. The mixture was stirred for 2 hours, evaporated to dryness, and extracted with ethyl acetate and 30 _mL of water. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to give 1.32 g of a yellow solid. Yield: 100.0%.

(9) Preparation of (5-ethynyl-1-methyl-1H-pyrrole-2-yl)methanol

Methyl 5-ethynyl-1-methyl-1H-pyrrole-2-carboxylate (1.05 g, 6.43 mmol) was dissolved in 20 mL of anhydrous THF. _LiAlH₄ (0.367 g, 9.66 mmol) was slowly added at 0 °C. The reaction was stirred at room temperature for 7 hours, cooled to 0 °C, and ethyl acetate was added until no more bubbles were produced. 2 mL of water was added, and the mixture was filtered. The filtrate was extracted with ethyl acetate, washed with 20 mL of water, dried over anhydrous sodium sulfate, and evaporated to dryness. Silica gel column chromatography (100% petroleum ether → petroleum ether:ethyl acetate = 2:1) was performed to give 0.67 g of a red solid, yield: 77.1%.

Preparation of (10) (2S,3R)-3-hydroxy-2-(4-((5-(hydroxymethyl)-1-methyl-1H-pyrrolo-2-yl)but-1,3-diynyl)benzoylaminobutyrate methyl ester

CuCl (3 mg, 0.03 mmol) and hydroxylamine hydrochloride (6 mg, 0.09 mmol) were dissolved in 10 mL of 23% n-butylamine aqueous solution. Then, 5 mL of methanol and tetrahydrofuran solution (V:V, 1:1) containing (5-ethynyl-1-methyl-1H-pyrrole-2-yl)methanol (0.217 g, 1.60 mmol) and 5 mL of methanol and tetrahydrofuran solution (V:V, 1:1) containing (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate (0.496 g, 1.46 mmol) were added to the above reaction solution. The mixture was stirred for 2 min, then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. Silica gel column chromatography (100% petroleum ether → petroleum ether:ethyl acetate = 2:1) yielded 0.54 g of a pale red solid, yield: 93.8%.

(11) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-(hydroxymethyl)-1-methyl-1H-pyrrole-2-yl)but-1,3-diynyl)benzoamide

Methyl (2S,3R)-3-hydroxy-2-(4-((5-(hydroxymethyl)-1-methyl-1H-pyrrolo-2-yl)but-1,3-diynyl)benzoylaminobutyrate (0.50 g, 1.27 mmol) was dissolved in 10 mL of methanol, and 2 mL of 50% hydroxylamine aqueous solution and lithium hydroxide monohydrate (0.018 g, 0.43 mmol) were added. The reaction mixture was stirred for 48 hours, evaporated to dryness, and purified by liquid chromatography to obtain 0.178 g of compound 1, yield: 35.4%.

_{Molecular formula: C₂₁H₂₁N₃O₅ Molecular weight} : _395.1 Mass spectrometry: (M+H): 396.1

^1H -NMR(d ₆ -DMSO, 400MHz) δ 10.66 (1H, s), 8.83 (1H, s), 8.16 (1H, d), 7.92 (2H, d), 7.68 (2H, d), 6.57 (1H, d), 6.02 (1H, d), 5.12 (1H, t), 4.89 (1H, br s), 4.41(2H, d), 4.24(1H, dd), 4.01(1H, t), 3.63(3H, s), 1.07(3H, d).

Example 28 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((1-methyl-1H- Preparation of pyrazol-5-yl)but-1,3-diynyl)benzamide (compound 34)

Referring to Example 28, compound 34 was synthesized.

_Molecular formula: _{C19H18N4O4 Molecular} weight: _366.1 Mass spectrometry (M+H): 367.1

^1H -NMR(d ₆ -DMSO, 400MHz) δ10.69 (1H, s), 8.85 (1H, s), 8.23 (1H, d), 7.97 (2H, d), 7.76 (2H, d), 7.5 6(1H,d), 6.79(1H,d), 4.92(1H,m), 4.26(1H,d), 4.03(1H,m), 3.94(3H,s), 1.09(3H,d).

Example 29 4-((6-(furan-2-yl)pyridin-3-yl)ethynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxylamine) Preparation of (-1-oxobutane-2-yl)benzamide (compound 35)

(1) Preparation of (S)-3-methyl-3-nitro-2-((S)-1-phenylethylamino)butyric acid

2-Nitropropane (18.5 mL, 0.206 mol) and potassium hydroxide (13.6 g, 0.242 mol) were dissolved in 200 mL of water. Under nitrogen protection, the solution was heated to 45 °C, and S-1-phenylethylamine (25 g, 0.206 mol) was rapidly added. Then, 50% glyoxylic acid aqueous solution (29.92 g) was slowly added dropwise. After the addition was complete, the solution was stirred at 35 °C for 3 h. Then, 3M dilute hydrochloric acid (152 mL, 0.456 mol) was slowly added dropwise, and the solution was stirred overnight at room temperature. A white precipitate formed. The precipitate was filtered, and the solid was washed successively with dilute hydrochloric acid (0.2 M, 0.5 L), water (0.5 L), and diethyl ether (0.125 L). After drying, 27.43 g of a white powdery solid was obtained, with a yield of 50%.

(2) Preparation of methyl (S)-3-methyl-3-nitro-2-((S)-1-phenylethylamino)butyrate

(S)-3-methyl-3-nitro-2-((S)-1-phenylethylamino)butyric acid (25 g, 93.9 mmol) and cesium carbonate (33.6 g, 103 mmol) were added to N,N-dimethylformamide (100 mL). Iodomethane (15.32 g, 108 mmol) was added dropwise at 0 °C under nitrogen protection. After the addition was complete, the mixture was allowed to react at room temperature for 12 h. The solution was poured into water, the pH was adjusted to approximately 7-8 with dilute hydrochloric acid, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain 26.3 g of an oily substance, which was used directly in the next step without purification.

(3) Preparation of methyl (S)-3-amino-3-methyl-2-((S)-1-phenylethylamino)butyrate

The crude product obtained in the previous step (26.3 g, approximately 93.9 mmol) was dissolved in tetrahydrofuran (100 mL) and glacial acetic acid (150 mL). Zinc powder (total 55.3 g, 845 mmol) was added in multiple batches under ice bath conditions. The mixture was then reacted at room temperature for 18 h. After filtration, the filter cake was washed with tetrahydrofuran. The filtrate was concentrated, and water was added. The pH was adjusted to approximately 9-10 with saturated sodium bicarbonate. The mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated to obtain 23.5 g of an oily substance, which was used directly in the next step without purification.

(4) Preparation of methyl (S)-3-(tert-butoxycarbonylamino)-3-methyl-2-((S)-1-phenylethylamino)butyrate

The crude product (23.5 g), diisopropylethylamine (24.3 g, 188 mmol), and Boc-anhydride (22.5 g, 103 mmol) obtained in the previous step were dissolved in tetrahydrofuran (150 mL), stirred at room temperature for 16 h, concentrated, and then dissolved in ethyl acetate. The solution was washed successively with 10% citric acid, semi-saturated sodium bicarbonate solution, and brine, dried over anhydrous sodium sulfate, concentrated, and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 13.2 g of a transparent oil.

(5) Preparation of methyl (S)-2-amino-3-(tert-butoxycarbonylamino)-3-methylbutyrate

Methyl (S)-3-(tert-butoxycarbonylamino)-3-methyl-2-((S)-1-phenylethylamino)butyrate (5.2 g, 14.8 mmol) was dissolved in tetrahydrofuran (150 mL), and 20% palladium hydroxide on carbon (1.8 g) was added. The mixture was reacted under a hydrogen atmosphere for 4 h, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (dichloromethane:methanol = 10:1) to give 3.35 g, with a yield of 91.9%.

(6) Preparation of 5-(4-iodophenyl)-1-methyl-1H-pyrazole

1-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole (833 mg, 4.0 mmol), p-diiodobenzene (1.497 g, 4.54 mmol), tetrakis(triphenylphosphine)palladium (462 mg, 0.4 mmol), and sodium carbonate (636 mg, 6.0 mmol) were dissolved in 1,4-dioxane (30 mL) and water (10 mL), and reacted at 90 °C for 6 h under nitrogen protection. After cooling, the solution was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 633 mg of a white solid, in 55.8% yield.

(7) Preparation of methyl (S)-3-(tert-butoxycarbonylamino)-2-(4-ethynylbenzoylamino)-3-methylbutyrate

Methyl (2S,3R)-3-hydroxy-2-(4-ethynylbenzamido)butyrate (439 mg, 3.0 mmol), methyl (S)-2-amino-3-(tert-butoxycarbonylamino)-3-methylbutyrate (887 mg, 3.6 mmol), EDCI (691 mg, 3.6 mmol), HOBt (487 mg, 3.6 mmol), and N,N-diisopropylethylamine (1.55 g, 12.0 mmol) were dissolved in N,N-dimethylacetamide (30 mL) and reacted at room temperature for 12 h. The solution was poured into water, extracted with ethyl acetate, and the organic phase was concentrated and then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to give 1.1 g of a yellow solid, yield 98%.

(8) Preparation of (S)-3-(tert-butoxycarbonylamino)-3-methyl-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)methyl butyrate

5-(4-iodophenyl)-1-methyl-1H-pyrazole (396 mg, 1.39 mmol), (S)-3-(tert-butoxycarbonylamino)-2-(4-ethynylbenzoylamino)-3-methylbutyrate (522 mg, 1.39 mmol), bis(triphenylphosphine)palladium dichloride (20 mg, 0.028 mmol), cuprous iodide (3 mg, 0.016 mmol), and triethylamine (423 mg, 4.18 mmol) were dissolved in acetonitrile (30 mL), and the reaction was carried out at room temperature for 3 h under nitrogen protection. The solution was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give 689 mg of a white solid, yield 93.4%.

(9) Preparation of (S)-3-(tert-butoxycarbonylamino)-3-methyl-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)butyric acid

Methyl (S)-3-(tert-butoxycarbonylamino)-3-methyl-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)butyrate (354 mg, 0.667 mmol) and lithium hydroxide monohydrate (84 mg, 2.0 mmol) were dissolved in a mixed solvent of tetrahydrofuran and water (30 mL, v/v = 4:1) and reacted at room temperature for 8 h. The pH was adjusted to 6 with dilute hydrochloric acid under an ice-water bath, and the precipitate was obtained. After filtration, 317 mg of a white solid was obtained, with a yield of 92.1%.

(10) Preparation of (3S)-2-methyl-3-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)-4-oxo-4-(tetrahydro-2H-pyran-2-yloxyamino)butane-2-ylcarbamate tert-butyl ester

(S)-3-(tert-butoxycarbonylamino)-3-methyl-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)butyric acid (317 mg, 0.614 mmol), O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (144 mg, 1.23 mmol), HATU (350 mg, 0.920 mmol), and triethylamine (187 mg, 1.84 mmol) were dissolved in DMA (30 mL), stirred at room temperature for 6 h, poured into water, extracted with ethyl acetate, concentrated the organic phase, and then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to give 105 mg of yellow solid, yield 27.9%.

(11) Preparation of (S)-N-(3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)-4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamide

(3S)-2-methyl-3-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)-4-oxo-4-(tetrahydro-2H-pyran-2-yloxyamino)butane-2-ylcarbamate tert-butyl ester (105 mg, 0.171 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (5 mL) was added dropwise in an ice bath. The reaction was carried out at room temperature for 2 h. After concentration, the mixture was purified by preparative liquid chromatography to give 25 mg of white solid, with a yield of 33.9%.

_Molecular formula: _{C₂₄H₂₅N₅O₃ Molecular} weight: _431.2 Mass spectrometry (M+H): 432.1

^1H -NMR(d ₆ -DMSO, 400MHz) δ 8.42-8.25 (3H, m), 7.93 (2H, d), 7.74-7.65 (4H, m), 7.62 (2H, d) , 7.48(1H,d), 6.49(1H,d), 4.35(1H,s), 3.89(3H,s), 1.13(3H,s), 1.05(3H,s).

Example 30 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((4-(1-methyl- Preparation of 1H-pyrazole-5-yl)phenyl)ethynyl)benzamide (compound 36)

(1) Preparation of methyl (2S,3R)-2-(4-((4-bromophenyl)ethynyl)benzamido)-3-hydroxybutyrate

Methyl (2S,3R)-2-(4-ethynylbenzamido)-3-hydroxybutyrate (1.00 g, 3.83 mmol), p-bromoiodobenzene (1.48 g, 5.23 mmol), triethylamine (0.893 g, 8.82 mmol), Pd( _PPh3 ) _2Cl2 ( _0.094 g, 0.134 mmol), and CuI (0.047 g, 0.247 mmol) were dissolved in 20 mL of THF. The mixture was purged with nitrogen three times, stirred at room temperature for 18 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 1:2) to give 1.1 g of white solid, yield 68.9%.

(2) Preparation of (2S,3R)-3-hydroxy-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamide)butyric acid

Methyl (2S,3R)-2-(4-((4-bromophenyl)ethynyl)benzamido)-3-hydroxybutyrate (1.10 g, 2.64 mmol), 1-methyl-1H-pyrazole-5-boronic acid pinacol ester (0.66 g, 3.17 mmol), sodium carbonate (0.421 g, 3.97 mmol), and Pd( _PPh3 ) ₄ (0.306 g, 0.265 mmol) were dissolved in 20 mL of 1,4-dioxane and 1 mL of water. The mixture was purged with nitrogen three times, refluxed and stirred for 18 hours, evaporated to dryness, and purified by silica gel column chromatography (100% dichloromethane → dichloromethane:methanol = 10:1) to give 0.821 g of a colorless oily product, with a yield of 77.1%.

(3) Preparation of (2S,3R)-3-hydroxy-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamide)methyl butyrate

(2S,3R)-3-hydroxy-2-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamido)butyric acid (0.452 g, 1.12 mmol) was dissolved in 15 mL of methanol, and then 3 mL of thionyl chloride was added at 0 °C. The mixture was stirred for 2 hours and evaporated to dryness to obtain a white solid, which was used directly in the next step.

(4) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamide

The solid obtained in the previous step was dissolved in 10 mL of methanol, and 8 mL of 50% hydroxylamine aqueous solution and lithium hydroxide monohydrate (0.032 g, 0.762 mmol) were added. The reaction was stirred for 3 hours, and a white solid precipitated out. The mixture was filtered, and the filter cake was washed with methanol and water, respectively, to obtain 0.096 g of white solid. The yield of the two steps was 20.4%.

_{Molecular formula: C₂₃H₂₂N₄O₄ Molecular weight} : _418.2 Mass spectrometry (M+H): 419.2

^1H -NMR(d ₆ -DMSO, 400MHz) δ10.72 (1H, s), 8.85 (1H, s), 8.17 (1H, d), 7.96 (2H, d), 7.80-7.60 (6H, m), 7.49 (1H, d), 6.49 (1H, d), 5.15-4.80 (1H, br), 4.26 (1H, q), 4.03 (1H, t), 3.89 (3H, s), 1.07 (3H, d).

Example 31 (S)-N-(3-amino-1-(hydroxyamino)-3-methyl-1-oxobutane-2-yl)-4-((4-(3- Preparation of (hydroxymethyl)-1-methyl-1H-pyrazole-5-yl)phenyl)ethynyl)benzamide (compound 37)

Preparation of (1) methyl (S)-3-(tert-butoxycarbonylamino)-2-(4-((4-(3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamido)-3-methylbutyrate

Methyl (S)-3-(tert-butoxycarbonylamino)-2-(4-ethynylbenzoylamino)-3-methylbutyrate (0.30 g, 0.801 mmol), (5-(4-iodophenyl)-1-methyl-1H-pyrazol-3-yl)methanol (0.252 g, 0.801 mmol), triethylamine (0.242 g, 2.40 mmol), Pd( _PPh3 ) _{2Cl2 (} 0.022 g, 0.031 mmol), and CuI (3 mg, 0.016 mmol) were dissolved in 20 mL of acetonitrile. The mixture was purged with nitrogen three times, stirred at room temperature for 17 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% dichloromethane → dichloromethane:methanol = 20:1) to give 0.320 g of white solid, yield 71.3%.

(2) Preparation of (S)-3-amino-2-(4-((4-(3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzoylamino)-3-methylbutyrate methyl ester

Methyl (S)-3-(tert-butoxycarbonylamino)-2-(4-((4-(3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamido)-3-methylbutyrate (0.30 g, 0.535 mmol) was dissolved in 10 mL of dichloromethane, 1 mL of TFA was added, the reaction was stirred for 2 h, and the solution was evaporated to dryness to obtain 0.36 g of crude product, a brownish-red oily substance, which was directly used in the next step.

(3) Preparation of (S)-N-(3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)-4-((4-(3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)benzamide

The oily substance obtained in the previous step was dissolved in 10 mL of methanol, and 2 mL of 50% hydroxylamine aqueous solution and lithium hydroxide monohydrate (0.013 g, 0.31 mmol) were added. The reaction was stirred for 70 hours, evaporated to dryness, and liquid-phase purification was prepared (100% water → water:methanol = 100:62) to obtain 0.066 g of white solid. The yield of the two steps was 26.7%.

_Molecular formula: _{C₂₅H₂₇N₅O₄ Molecular} weight: _461.2 Mass spectrometry (M+H): 462.2

^1H -NMR(d ₆ -DMSO, 600MHz) δ 8.12 (1H, s), 7.88 (2H, d), 7.79 (2H, d), 7.62 (2H, d), 7.56 (2H, d), 6.66 (1H, s), 5 .31(1H,m),5.15-4.68(2H,br),4.50(2H,d),4.17(1H,s),3.82(3H,s),1.07(3H,s),0.99(3H,s).

Example 32 N-hydroxy-2-methyl-4-(4-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl) Preparation of phenyl)-2-oxopyridine-1(2H)-yl)-2-(methanesulfonyl)butyramide (compound 38)

(1) Preparation of 3-(dimethylamino)-1-(4-iodophenyl)prop-2-enyl-1-one

4-Iodoacetophenone (4.92 g, 20.00 mmol) and N,N-dimethylformamide dimethyl acetal (4.77 g, 40.00 mmol) were dissolved in N,N-dimethylformamide (15.00 mL) and reacted at 125 °C for 3 h. The mixture was then concentrated under reduced pressure to obtain 6.02 g of a red oily substance, which was used directly in the next step.

(2) Preparation of 5-(4-iodophenyl)-1-methyl-1H-pyrazole

3-(dimethylamino)-1-(4-iodophenyl)prop-2-enyl-1-one (6.02 g, 20.00 mmol) was dissolved in N,N-dimethylformamide (10.00 mL), and methylhydrazine (2.76 g, 60.00 mmol) was added. The mixture was reacted at room temperature for 1 h, and then at 75 °C for 4 h. The mixture was concentrated under reduced pressure and subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 3.35 g of solid, with a yield of 58.9%.

(3) Preparation of 1-methyl-5-(4-((trimethylsilyl)ethynyl)phenyl)-1H-pyrazole

5-(4-iodophenyl)-1-methyl-1H-pyrazole (2.60 g, 9.15 mmol), trimethylsilylacetylene (1.79 g, 18.30 mmol), bis(triphenylphosphine)palladium dichloride (323 mg, 0.46 mmol), cuprous iodide (88 mg, 0.46 mmol), and triethylamine (2.78 g, 27.45 mmol) were dissolved in 1,4-dioxane (40 mL) and refluxed for 3 h under nitrogen protection. After cooling and concentration, 2.33 g of crude solid was directly used in the next step.

(4) Preparation of 5-(4-ethynylphenyl)-1-methyl-1H-pyrazole

The crude product (2.33 g, 9.15 mmol) from the previous step was dissolved in methanol (50 mL), and potassium carbonate (2.53 g, 18.30 mmol) was added. The reaction was carried out at room temperature for 4 hours. After the reaction was completed, 100 mL of water was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness. Column chromatography (PE:EA = 5:1) was performed to give 1.47 g of white solid, with a yield of 88.5%.

(5) Preparation of 1-methyl-5-(4-((4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl)ethynyl)phenyl)1H-pyrazole

5-(4-ethynylphenyl)-1-methyl-1H-pyrazole (273 mg, 1.50 mmol), pinacol 4-iodophenylboronic acid (495 mg, 1.50 mmol), palladium dichloride bis(triphenylphosphine) (21 mg, 0.030 mmol), cuprous iodide (3 mg, 0.016 mmol), and triethylamine (455 mg, 4.50 mmol) were dissolved in acetonitrile (15 mL), and the reaction was carried out at room temperature for 6 h under nitrogen protection. After cooling, the solution was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to give 424 mg of a white solid, in 73.6% yield.

(6) Preparation of 2-methyl-4-(4-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)phenyl)-2-oxopyridin-1(2H)-yl)-2-(methanesulfonyl)-N-(tetrahydro-2H-pyran-2-yloxy)butyramide

The following ingredients were added: 1-methyl-5-(4-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl)ethynyl)phenyl)-1H-pyrazole (234 mg, 0.61 mmol), 4-(4-iodo-2-oxopyridin-1(2H)-yl)-2-methyl-2-(methanesulfonyl)-N-(tetrahydro-2H-pyran-2-yloxy)butyramide (304 mg, 0.61 mmol), and [1,1′-bis(diphenylphosphine)]. Ferrocene]palladium(II) dichloromethane complex (50 mg, 0.061 mmol) and potassium phosphate (388 mg, 1.83 mmol) were added to 1,4-dioxane (20 mmol) and water (1 mL). The reaction was carried out at 90 °C for 16 h under nitrogen protection. After cooling, the mixture was concentrated and dissolved in ethyl acetate. The solution was washed with water, dried over anhydrous sodium sulfate, concentrated, and subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to give 177 mg of solid, yield 46.3%.

(7) Preparation of N-hydroxy-2-methyl-4-(4-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)phenyl)-2-oxopyridin-1(2H)-yl)-2-(methanesulfonyl)butyramide

177 mg (0.28 mmol) of 2-methyl-4-(4-(4-((4-(1-methyl-1H-pyrazol-5-yl)phenyl)ethynyl)phenyl)-2-oxopyridin-1(2H)-yl)-2-(methanesulfonyl)-N-(tetrahydro-2H-pyran-2-yloxy)butyramide was dissolved in 20 mL of dichloromethane. Trifluoroacetic acid (3 mL) was added dropwise in an ice bath, and the reaction was carried out at room temperature for 5 h. TLC (petroleum ether:ethyl acetate = 1:1) showed that the reaction was complete. After concentration under reduced pressure, the solution was washed with a small amount of diethyl ether and acetonitrile to give 58 mg of white solid, with a yield of 38.1%.

_Molecular formula: C₂ _{₹ H₂₈N₄O₅S} Molecular weight: _544.2 Mass spectrometry (M+H): 545.2

^1H -NMR(d ₆ -DMSO, 400MHz) δ7.82 (2H, d), 7.77 (1H, d), 7.73-7.67 (4H, m), 7.63 (2H, d), 7.50 (1H, d), 6.79-6.73 (1H, m), 6.69 (1H, d), 6.5 0(1H,d), 4.18-4.10(1H,m), 3.90(3H,s), 3.77-3.71(1H,m), 3.10(3H,s), 2.46-2.37(1H,m), 2.18-2.10(1H,m), 1.52(3H,s).

Example 33 4-((4-(1H-pyrazol-5-yl)phenyl)ethynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxylamine) Preparation of (-1-oxobutane-2-yl)benzamide (compound 39)

(1) Preparation of (2S,3R)-3-hydroxy-2-(4-((4-iodophenyl)ethynyl)benzamido)methyl butyrate

Methyl (2S,3R)-2-(4-ethynylbenzamido)-3-hydroxybutyrate (1.00 g, 3.83 mmol), 1,4-diiodobenzene (1.726 g, 5.23 mmol), triethylamine (0.893 g, 8.82 mmol), Pd( _PPh3 ) _2Cl2 ( _0.094 g, 0.134 mmol), and CuI (0.047 g, 0.247 mmol) were dissolved in 20 mL of THF, purged three times with nitrogen, stirred at room temperature for 18 hours, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 1:2) to give 1.3 g of white solid, yield 73.4%.

(2) Preparation of methyl (2S,3R)-2-(4-((4-(1H-pyrazol-5-yl)phenyl)ethynyl)benzamido)-3-hydroxybutyrate

Methyl (2S,3R)-3-hydroxy-2-(4-((4-iodophenyl)ethynyl)benzamido)butyrate (0.334 g, 0.721 mmol), 1H-pyrazole-5-boronic acid (0.097 g, 0.867 mmol), sodium carbonate (0.115 g, 1.085 mmol), and Pd( _PPh3 ) ₄ (0.041 g, 0.035 mmol) were dissolved in 10 mL of 1,4-dioxane and 2 drops of water. The mixture was purged with nitrogen three times, refluxed and stirred for 18 hours, evaporated to dryness, and purified by silica gel column chromatography (100% dichloromethane → dichloromethane:methanol = 10:1) to give 0.12 g of a colorless oily product, yield 41.2%.

(3) Preparation of 4-((4-(1H-pyrazol-5-yl)phenyl)ethynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)benzamide

Methyl (2S,3R)-2-(4-((4-(1H-pyrazol-5-yl)phenyl)ethynyl)benzamido)-3-hydroxybutyrate (0.12 g, 0.297 mmol) was dissolved in 10 mL of methanol, and 8 mL of 50% hydroxylamine aqueous solution and lithium hydroxide monohydrate (0.008 g, 0.19 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours, evaporated to dryness, and purified by silica gel column chromatography to give 0.056 g of compound 37, yield: 46.5%.

_Molecular formula: _{C₂₂H₂ON₄O₄ Molecular} weight: _404.1 Mass spectrometry (M+H): 405.2

^1H -NMR(d ₆ -DMSO, 400MHz) δ13.01 (1H, s), 10.69 (1H, s), 8.86 (1H, s), 8.15 (1H, d), 7.95 (2H, d), 7.89 (2H, d), 7.8 1(1H,m), 7.66(2H,d), 7.60(2H,d), 6.80(1H,m), 4.90(1H,d), 4.25(1H,dd), 4.02(1H,q), 1.08(3H,d).

Example 34 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((6-(1-methyl- Preparation of 1H-pyrazol-5-yl)pyridin-3-yl)ethynyl)benzamide (compound 40)

(1) Preparation of 5-bromo-2-(1-methyl-1H-pyrazol-5-yl)pyridine

In a dry reaction flask, 2,5-dibromopyridine (521 mg, 2.2 mmol) and 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (416 mg, 2.0 mmol) were dissolved in a mixed solvent of 8 mL toluene and 8 mL ethanol. 4 mL of an aqueous solution of sodium carbonate (636 mg, 6.0 mmol) and Pd( _PPh3 ) ₄ (69 mg, 0.06 mmol) were added. After the addition was complete, nitrogen was introduced, and the reaction was stirred at 60 °C for 12 hours. The mixture was then cooled to room temperature, and the organic phase was concentrated under reduced pressure. Column chromatography (PE:EA = 20:1) yielded 400 mg of a pale yellow solid, with a yield of 84%.

(2) Preparation of (2S,3R)-3-hydroxy-2-(4-((6-(1-methyl-1H-pyrazol-5-yl)pyridin-3-yl)ethynyl)benzamido)methyl butyrate

In a dry reaction flask, 5-bromo-2-(1-methyl-1H-pyrazol-5-yl)pyridine (66 mg, 0.277 mmol) and (2S,3R)-2-(4-ethynylbenzamido)-3-hydroxybutyrate methyl ester (72 mg, 0.277 mmol) were added and dissolved in 5 mL acetonitrile and 3 mL triethylamine. Copper iodide (0.5 mg, 0.0027 mmol) and Pd( _PPh3 ) _2Cl2 (4 mg, 0.0054 mmol) were added _. The air was replaced with a small amount of a mixture of hydrogen and nitrogen. The mixture was stirred at 80 °C for 2.5 h, cooled to room temperature, and the organic phase was concentrated under reduced pressure. Column chromatography (PE:EA = 1:2) yielded 85 mg of a pale yellow solid, with a yield of 73.3%.

(3) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((6-(1-methyl-1H-pyrazol-5-yl)pyridin-3-yl)ethynyl)benzamide

In a dry reaction flask, methyl (2S,3R)-3-hydroxy-2-(4-((6-(1-methyl-1H-pyrazol-5-yl)pyridin-3-yl)ethynyl)benzamido)butyrate (85 mg, 0.203 mmol) was added and dissolved in 4 mL of methanol. Then, 2 mL of 50% hydroxylamine aqueous solution was added, and the mixture was stirred at room temperature for 1 hour. Lithium hydroxide monohydrate (10 mg, 0.238 mmol) was added, and the mixture was stirred at room temperature for 12 hours after the addition was complete. Water was added, and a solid precipitated. The solid was filtered, washed with water, and dried to obtain 68 mg of a white solid, with a yield of 79.9%.

_{Molecular formula: C₂₂H₂₁N₅O₄ Molecular weight} : _419.2 Mass spectrometry (M+H): 420.2

¹ H-NMR (d ₆ -DMSO, 400MHz): δ 8.87 (1H, s), 8.10 (1H, d), 7.96-7.82 (4H, m), 7.69 (2H, d), 7.51 (1H, d), 6.90 (1H, s), 6.41 (1H, s), 5.16 (1H, br s), 4.28-4.22(1H, m), 4.17(3H, s), 4.02-3.88(1H, m), 1.01(3H, d).

Example 35 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(hydroxymethyl)) Preparation of thiophene-2-yl)but-1,3-diynyl)benzamide (compound 41)

(1) Preparation of 5-((trimethylsilyl)ethynyl)thiophene-2-carboxaldehyde

5-Bromothiophene-2-carboxaldehyde (2.7 g, 14.1 mmol), trimethylsilylacetylene (2.1 g, 21.4 mmol), triethylamine (2.88 g, 28.6 mmol), Pd( _PPh3 ) _2Cl2 (100 mg, 0.14 mmol), and CuI (27 _mg , 0.14 mmol) were dissolved in 20 mL of THF. The mixture was purged with nitrogen three times, stirred at room temperature for 3 hours, extracted with ethyl acetate, separated, evaporated to dryness, and subjected to silica gel column chromatography (100% petroleum ether → petroleum ether: ethyl acetate = 20:1) to give 2.0 g of a pale yellow solid, yield: 68.1%.

(2) Preparation of (5-((trimethylsilyl)ethynyl)thiophen-2-yl)methanol

5-((trimethylsilyl)ethynyl)thiophene-2-carboxaldehyde (624 mg, 2.99 mmol) was dissolved in 10 mL of THF, and _NaBH4 (228 mg, 6.0 mmol) was added. The mixture was stirred for 0.5 hours, evaporated to dryness, and used directly in the next step of the reaction.

(3) Preparation of (5-ethynylthiophene-2-yl)methanol

The crude product obtained in the previous step was added to 10 mL of anhydrous methanol, and _K2CO3 (828 mg, 6.0 mmol) was _added . The reaction was stirred at room temperature for 16 hours. The system was filtered, the filtrate was evaporated to dryness, and silica gel column chromatography (PE:EA = 10:1-5:1) was performed to obtain 280 mg of oil. The two-step yield was 67.9%.

(4) Preparation of (2S,3R)-3-hydroxy-2-(4-((5-(hydroxymethyl)thiophen-2-yl)but-1,3-diynyl)benzoylamino)methyl butyrate

CuCl (4 mg, 0.04 mmol) and hydroxylamine hydrochloride (14 mg, 0.2 mmol) were dissolved in 5 mL of 23% n-butylamine aqueous solution. 2.5 mL of 23% n-butylamine aqueous solution containing (5-ethynylthiophene-2-yl)methanol (280 mg, 2.03 mmol) was added. Then, 2.5 mL of methanol and 1.0 mL of tetrahydrofuran solution containing (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate (400 mg, 1.18 mmol) were added to the above reaction solution. The mixture was stirred for 5 min, then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. Silica gel column chromatography (PE:EA = 10:1–2:3) yielded 200 mg of the product, yield: 42.6%.

(5) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-(hydroxymethyl)thiophene-2-yl)but-1,3-diynyl)benzamide

Methyl (2S,3R)-3-hydroxy-2-(4-((5-(hydroxymethyl)thiophen-2-yl)but-1,3-diynyl)benzoylamino)butyrate (200 mg, 0.503 mmol) was dissolved in 2 mL of methanol, and 2 mL of 50% hydroxylamine aqueous solution and lithium hydroxide monohydrate (9 mg, 0.214 mmol) were added. The reaction was stirred for 72 hours, methanol was removed by rotary evaporation, and the mixture was filtered. The filter cake was dissolved in DMSO, and the product was purified by preparative liquid chromatography to obtain 40 mg of product, yield: 19.9%.

_{Molecular formula: C₂₀H₁₈N₂O₅S Molecular weight} : _398.1 Mass spectrometry (M+H): 398.9

^1H -NMR(d ₆ -DMSO, 400MHz) δ 10.66 (1H, s), 8.84 (1H, s), 8.18 (1H, d), 7.93 (2H, d), 7.70 (2H, d), 7.44 (1H, d), 6.9 5(1H,d), 5.71(1H,t), 4.93-4.84(1H,m), 4.65(2H,d), 4.24(1H,dd), 4.06-3.96(1H,m), 1.07(3H,d).

Example 36 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-((2-(2- Preparation of hydroxyethoxy)ethylamino)methyl)thiophene-2-yl)but-1,3-diynyl)benzamide (compound 42)

(1) Preparation of 2-(2-((5-((trimethylsilyl)ethynyl)thiophen-2-yl)methyleneamino)ethoxy)ethanol

Dissolve 5-((trimethylsilyl)ethynyl)thiophene-2-carboxaldehyde (0.433 g, 2.08 mmol) in 10 mL of methanol, add 2-(2-aminoethoxy)ethanol (0.218 g, 2.08 mmol), add 1 drop of acetic acid, stir overnight at room temperature, and use directly in the next reaction step.

(2) Preparation of 2-(2-((5-((trimethylsilyl)ethynyl)thiophen-2-yl)methylamino)ethoxy)ethanol

Sodium borohydride (0.236 g, 6.24 mmol) was added to the reaction system of the previous step, and the mixture was stirred at room temperature for 1 hour. LC-MS showed that the reaction was complete and no further processing was required. The mixture was then used directly for the next reaction.

(3) Preparation of 2-(2-((5-ethynylthiophene-2-yl)methylamino)ethoxy)ethanol

Potassium carbonate (0.862 g, 6.24 mmol) was added to the reaction system of the previous step, and the reaction was carried out at room temperature for 4 hours. After the reaction was completed, 100 mL of water was added to the system, and the organic phases were extracted three times with ethyl acetate. The combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to column chromatography (PE∶EA＝5∶1→DCM∶MeOH＝20∶1) to give 0.25 g of brown oil. The yield of the three steps was 53.4%.

(4) Preparation of (2S,3R)-3-hydroxy-2-(4-((5-((2-(2-hydroxyethoxy)ethylamino)methyl)thiophen-2-yl)but-1,3-diynyl)benzoylamino)methyl butyrate

CuCl (6 mg, 0.06 mmol) and hydroxylamine hydrochloride (14 mg, 0.2 mmol) were dissolved in 6 mL of 23% n-butylamine aqueous solution. Then, 5 mL of methanol and tetrahydrofuran solution (V:V, 1:1) containing 2-(2-((5-ethynylthiophene-2-yl)methylamino)ethoxy)ethanol (0.25 g, 1.11 mmol) were added sequentially. A solution of methyl 3-hydroxybutyrate (0.377 g, 1.11 mmol) in 5 mL of methanol and tetrahydrofuran (V:V, 1:1) was added to the above reaction solution, stirred for 2 min, and then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted three times, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The solution was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1 → dichloromethane:methanol = 20:1) to give 0.22 g of a yellow solid, yield: 40.9%.

(5) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-((2-(2-hydroxyethoxy)ethylamino)methyl)thiophene-2-yl)but-1,3-diynyl)benzamide

Methyl (2S,3R)-3-hydroxy-2-(4-((5-(((2-(2-hydroxyethoxy)ethylamino)methyl)thiophene-2-yl)but-1,3-diynyl)benzoamide)butyrate (0.22 g, 0.454 mmol) was dissolved in 1 mL of methanol, and 4 mL of 50% hydroxylamine aqueous solution was added. The reaction was stirred for 1 hour, and the solution was directly purified by liquid phase (methanol:water = 40:100) to give 69 mg of yellow solid, yield: 31.3%.

_Molecular formula: _{C₂₄H₂₇N₃O₆S Molecular} weight: _485.2 Mass spectrometry (M+H): 486.2

¹ H-NMR (DMSO-d ₆ , 600MHz) δ10.67 (1H, s), 8.84 (1H, s), 8.18 (1H, d), 7.92 (2H, d), 7.68 (2H, d), 7.42 (1H, d), 6.95 (1H, d), 4.87 (1H, d), 4.6 3-4.52(1H,m), 4.22(1H,dd), 4.03-3.96(1H,m), 3.90(2H,s), 3.49-3.41(4H,m), 3.38(2H,t), 2.66(2H,t), 1.06(3H,d).

Example 37 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(morpholinomethyl) Preparation of (2-( ...

(1) Preparation of 4-((5-((trimethylsilyl)ethynyl)thiophen-2-yl)methyl)morpholine

5-((trimethylsilyl)ethynyl)thiophene-2-carboxaldehyde (458 mg, 2.2 mmol) and morpholine (174 mg, 2.0 mmol) were dissolved in 10 mL of DCM, and sodium triacetoxyborohydride (730 mg, 3.4 mmol) was added. The reaction was stirred at room temperature for 4 days (80 mg of sodium triacetoxyborohydride was added daily), and the solution was evaporated to dryness and used directly in the next step of the reaction.

(2) Preparation of 4-((5-ethynylthiophen-2-yl)methyl)morpholine

The crude product from the previous step was added to 10 mL of anhydrous methanol, _{and K₂CO₃} (828 mg, 6.0 mmol) was added. The reaction was stirred at room temperature for 16 hours. The system was filtered, the filtrate was evaporated to dryness, and silica gel column chromatography (PE∶EA＝10∶1→1∶1) was performed to obtain 200 mg of oil. The two-step yield was 48.2%.

(3) Preparation of (2S,3R)-3-hydroxy-2-(4-((5-(morpholinomethyl)thiophen-2-yl)but-1,3-diynyl)benzoylamino)methyl butyrate

CuCl (2 mg, 0.02 mmol) and hydroxylamine hydrochloride (7 mg, 0.1 mmol) were dissolved in 1.5 mL of 23% n-butylamine aqueous solution. Then, 0.5 mL of 23% n-butylamine aqueous solution containing 4-((5-ethynylthiophene-2-yl)methyl)morpholine (200 mg, 0.965 mmol) was added. Next, 1.5 mL of methanol and 0.75 mL of tetrahydrofuran solution containing (2S,3R)-2-(4-(bromoethynyl)benzoylamino)-3-hydroxybutyrate (200 mg, 0.588 mmol) were added to the above reaction solution. The mixture was stirred for 5 min, then 20 mL of ethyl acetate and 20 mL of water were added. The mixture was extracted, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The product was obtained by silica gel column chromatography (DCM:MeOH = 10:1), yielding 148 mg of product (yield: 53.9%).

(4) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-(morpholinomethyl)thiophene-2-yl)but-1,3-diynyl)benzamide

Methyl (2S,3R)-3-hydroxy-2-(4-((5-(morpholinomethyl)thiophen-2-yl)but-1,3-diynyl)benzoylamino)butyrate (148 mg, 0.317 mmol) was dissolved in 1 mL of methanol, and 3 mL of 50% hydroxylamine aqueous solution was added. The mixture was stirred at room temperature for 3 hours, and then directly purified by liquid phase (methanol:water = 50:100) to give 90 mg of the product, yield: 60.9%.

_Molecular formula: _{C₂₄H₂₅N₃O₅S Molecular} weight: _467.2 Mass spectrometry (M+H): 468.2

^1H -NMR(d ₆ -DMSO, 600MHz) δ 10.66 (1H, s), 8.82 (1H, s), 8.17 (1H, d), 7.92 (2H, d), 7.69 (2H, d), 7.43 (1H, d), 6.98 (1H, d), 4 .93-4.84(1H,m), 4.22(1H,dd), 4.00(1H,quintet), 3.68(2H,s), 3.56(4H,t), 2.42-2.36(4H,m), 1.06(3H,d).

Example 38 N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutane-2-yl)-4-((5-(pyridine-2-yl) Preparation of (2-thiophene-2-yl)ethynyl)benzamide (compound 44)

(1) Preparation of 2-(5-bromothiophene-2-yl)pyridine

A solution of bromine (5.45 g, 34.1 mmol) in dichloromethane (50 mL) was slowly added dropwise to a solution of 2-(thiophen-2-yl)pyridine (5 g, 31.0 mmol) in dichloromethane (50 mL) at 0 °C. After the addition was complete, the mixture was stirred at room temperature for 6 h. The solution was diluted with dichloromethane (200 mL) and washed successively with sodium bicarbonate solution, sodium sulfite solution, and saturated saline solution. The organic phase was dried over anhydrous sodium sulfate and concentrated to give 7.16 g of a red solid, with a yield of 96.2%.

(2) Preparation of (2S,3R)-3-(tert-butyldimethylsiloxy)-2-(4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamido)methyl butyrate

2-(5-bromothiophene-2-yl)pyridine (576 mg, 2.4 mmol), methyl (2S,3R)-3-(tert-butyldimethylsiloxy)-2-(4-ethynylbenzamido)butyrate (751 mg, 2.0 mmol), palladium dichloride bis(triphenylphosphine) (49 mg, 0.07 mmol), cuprous iodide (24 mg, 0.126 mmol), and triethylamine (607 mg, 6.0 mmol) were dissolved in tetrahydrofuran (60 mL), and the mixture was refluxed under nitrogen protection for 6 h. After cooling, the mixture was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give 710 mg of a white solid, in 66.4% yield.

(4) Preparation of (2S,3R)-3-(tert-butyldimethylsiloxy)-2-(4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamide)butyric acid

Methyl (2S,3R)-3-(tert-butyldimethylsiloxy)-2-(4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamido)butyrate (710 mg, 1.33 mmol) and lithium hydroxide monohydrate (279 mg, 6.64 mmol) were added to 33 mL of a tetrahydrofuran/methanol/water mixture (v/v/v = 5:5:1), and the reaction was carried out at room temperature for 18 h. The organic solvent was evaporated to dryness, water was added, the pH was adjusted to approximately 6 with dilute hydrochloric acid, the mixture was filtered, and dried to give 660 mg of a white solid, with a yield of 95.5%.

(5) Preparation of N-((2S,3R)-3-(tert-butyldimethylsiloxy)-1-oxo-1-(tetrahydro-2H-pyran-2-yloxyamino)butan-2-yl)-4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamide

(2S,3R)-3-(tert-butyldimethylsiloxy)-2-(4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamido)butyric acid (660 mg, 1.27 mmol), O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (223 mg, 1.90 mmol), 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (867 mg, 2.28 mmol), and triethylamine (257 mg, 2.54 mmol) were dissolved in N,N-dimethylformamide (15 mL), stirred at room temperature for 18 h, diluted with ethyl acetate, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to give 354 mg of white solid, yield 44.9%.

(6) Preparation of N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamide

N-((2S,3R)-3-(tert-butyldimethylsiloxy)-1-oxo-1-(tetrahydro-2H-pyran-2-yloxyamino)butan-2-yl)-4-((5-(pyridin-2-yl)thiophen-2-yl)ethynyl)benzamide (354 mg, 0.57 mmol) was dissolved in anhydrous ethanol (20 mL), and concentrated hydrochloric acid (5 mL) was slowly added dropwise at room temperature. The reaction was stirred for 6 h, concentrated under reduced pressure, and then purified by liquid chromatography (methanol/water = 20%) to obtain 180 mg of white solid, with a yield of 74.7%.

_Molecular formula: _{C₂₂H₁₉N₃O₄S Molecular} weight: _421.1 Mass spectrometry (M+H): 422.1

^1H -NMR(d ₆ -DMSO, 400MHz): δ10.70 (1H, s), 8.87 (1H, s), 8.54 (1H, d), 8.17 (1H, d), 8.03-7.75 (5H, m), 7 .67(2H,d), 7.50(1H,d), 7.33(1H,m), 4.92(1H,d), 4.25(1H,dd), 4.02(1H,q), 1.08(3H,d).

Referring to the above preparation method, different intermediates, D-allothreonine methyl ester hydrochloride (2R, 3R), L-allothreonine methyl ester hydrochloride (2S, 3S), and D-threonine methyl ester hydrochloride (2R, 3S), were used to prepare the final products with the corresponding configurations. Racemic threonine methyl ester hydrochloride can be used to prepare the racemic final product.

Example 39 N-hydroxy-2-methyl-2-(methanesulfonyl)-4-(4-(4-((5-(morpholinylmethyl)furan-2- Preparation of (compound 45) acetylenolphenyl 2-oxopyridine-1(2H)-yl butyramide

(1) Preparation of 4-((5-((4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethynyl)furan-2-yl)methyl)morpholine

4-((5-ethynylfuran-2-yl)methyl)morpholine (0.421 g, 2.20 mmol) was dissolved in 20 mL of acetonitrile, and p-iodophenylboronic acid pinacol ester (0.800 g, 2.42 mmol), cuprous iodide (0.021 g, 0.11 mmol), triethylamine (0.511 g, 5.06 mmol), and Pd( _Ph3 ) _2Cl2 ( _0.026 g, 0.037 mmol) were added. Under nitrogen protection, the mixture was reacted overnight at 45 °C. After the reaction was complete, the solution was evaporated to dryness and subjected to column chromatography (PE:EA = 20:1 → PE:EA = 2:1) to give 0.71 g of a brown solid, with a yield of 82.1%.

(2) Preparation of 2-methyl-2-(methanesulfonyl)-4-(4-(4-((5-(morpholinylmethyl)furan-2-yl)ethynyl)phenyl)-2-oxopyridin-1(2H)-yl)-N-(tetrahydro-2H-pyran-2-yloxy)butyramide

Dissolve 4-((5-((4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl)ethynyl)furan-2-yl)methyl)morpholine (0.359 g, 0.913 mmol) in 10 mL toluene, 2 mL ethanol, and 2 drops of water. Add potassium phosphate (0.484 g, 2.283 mmol) and Pd(dppf) _Cl₂. (0.056 g, 0.0761 mmol) was added to 4-(4-iodo-2-oxopyridin-1(2H)-yl)-2-methyl-2-(methanesulfonyl)-N-(tetrahydro-2H-pyran-2-yloxy)butyramide (0.379 g, 0.761 mmol). After the addition was complete, the mixture was stirred overnight in an oil bath at 80 °C. The organic phase was concentrated under reduced pressure and subjected to column chromatography (PE∶EA＝10∶1→EA) to give 0.15 g of a yellow oily substance, with a yield of 30.9%.

(3) Preparation of N-hydroxy-2-methyl-2-(methanesulfonyl)-4-(4-(4-((5-(morpholinylmethyl)furan-2-yl)ethynyl)phenyl)-2-oxopyridin-1(2H)-yl)butyramide

0.15 g (0.235 mmol) of 2-methyl-2-(methanesulfonyl)-4-(4-(4-((5-(morpholinylmethyl)furan-2-yl)ethynyl)phenyl)-2-oxopyridin-1(2H)-yl)-N-(tetrahydro-2H-pyran-2-yloxy)butyramide was dissolved in 5 mL of dichloromethane, then 2 mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 1 h. The pH was adjusted to 3 with dilute ammonia, and the mixture was evaporated to dryness. Liquid-phase purification (methanol:water = 45:100) was performed to obtain 78 mg of white solid, with a yield of 60.0%.

_Molecular formula: _{C₂₈H₃₁N₃O₇S Molecular} weight: _553.2 Mass spectrometry (M+H): 554.2

^1H -NMR(d ₆ -DMSO, 400MHz): δ11.18 (1H, s), 9.30 (1H, s), 7.83-7.67 (3H, m), 7.66 (2H, d), 6.89 (1H, d), 6.76 (1H, s), 6.70 (1H, d), 6.4 6(1H,d),4.12(1H,dt),3.75(1H,dt),3.60-3.40(10H,m),3.11(3H,s),2.45-2.30(1H,m),2.20-2.05(1H,m),1.58(3H,s)

Example 40 N-hydroxy-4-{4-[4-(5-hydroxymethyl-furan-2-ethyl)-phenyl]-2-oxo-2H-pyridine- Preparation of 1-yl}-2-methanesulfonyl-2-methylbutanamine (compound 46)

Referring to Example 39, compound 46 was synthesized.

_{Molecular formula: C₂₄H₂₄N₂O₇S Molecular} weight: _484.1 Mass spectrometry ₍ M+H): 485.2

¹ H-NMR (d ₆ -DMSO, 400MHz): δ _ppm 11.2(brs, 1H), 7.78(m, 3H), 7.63(d, 2H), 6.87(d, 1H), 6.75(s, 1H), 6.68(t, 1H), 6.42(s, 1H), 4.4 1(s, 2H), 4.1(m, 1H), 3.76(m, 2H), 3.10(s, 3H), 2.39-2.49(m, 1H), 2.14-2.20(m, 1H), 1.57(m, 3H).

Example 41 N-hydroxy-4-(4-{4-[4-(5-hydroxymethyl-furan-2-yl)-but-1,3-diynyl]-phenyl}- Preparation of 2-oxo-2H-pyridin-1-yl)-2-methanesulfonyl-2-methylbutanamine (compound 47)

Referring to Example 39, compound 47 was synthesized.

_{Molecular formula: C₂₆H₂₄N₂O₇S Molecular weight} : _508.13 Mass spectrometry (M+H): 509.1

¹ H-NMR (d ₆ -DMSO, 400MHz): δ _ppm 11.15(1H,s),7.83-7.79(3H,m),7.73(2H,d),7.07(1H,d),6.77(1H,d),6.69(1H,dd),6.45(1H,d),4.42(2H , s), 4.14-4.09 (1H, m), 3.7.8-3.74 (1H, m), 3.11 (3H, s), 2.46-2.41 (1H, m), 2.19-2.15 (1H, m), 1.57 (3H, s).

Example 42 N-1-hydroxy-2-methylsulfonyl-4-[4-[4-(5-methoxymethylfuran-2-ethyl)phenyl]- Preparation of 2-oxo-2H-pyridin-1-yl}-2-methylbutanediamine (compound 48)

Referring to Example 39, compound 48 was synthesized.

_{Molecular formula: C₂₅H₂₆N₂O₇S Molecular} weight: _498.1 Mass spectrometry ₍ M+H): 499.5

Industrial practicality

This invention provides an antibacterial agent that, as an LpxC inhibitor, exhibits excellent antibacterial activity against bacteria, particularly Gram-negative bacteria. Furthermore, the compound of this invention demonstrates excellent in vivo metabolic stability.

Claims (7)

1. Compounds of general formula (I), their pharmaceutically acceptable salts, deuterated derivatives, or stereoisomers thereof: in, W indicates ^R1 is selected from -( _CH2 ) _O-4C ( ^R1a , ^R1b )( _CH2 ) _O - ^4OR3 . Among them, ^R1a , ^R1b , and ^R3 are each independently selected from H, _C1-6 alkyl groups, and OH; R ^1' is selected from hydrogen and _C1-6 alkyl groups; _A1 , _A2 , and _A3 are each independently selected from CH; X and Y are each independently selected from alkynyl groups; ^R2 is B represents -( _C1-4 alkyl)-, C is selected from -OR ⁹ . ^R9 is selected from hydrogen atoms; m is 1. 2. The compound of claim 1, its pharmaceutically acceptable salt, deuterated derivative, or stereoisomer thereof: in, W indicates ^R1 is selected from -C( ^R1a , ^R1b ) ^OR3 , wherein ^R1a , ^R1b , and ^R3 are each independently selected from hydrogen atoms, methyl groups, and hydroxyl groups; R ^1' is selected from hydrogen and _C1-4 alkyl; _A1 , _A2 , and _A3 are each independently selected from CH; X and Y are each independently selected from alkynyl groups; ^R2 is B represents -( _C1-4 alkyl)-, C is selected from -OR ⁹ . ^R9 is selected from hydrogen atoms; m is 1. 3. The compound as described in claim 1 or 2, its pharmaceutically acceptable salt, deuterated form, or stereoisomer thereof: in, W indicates ^R1 is selected from -C(H, _CH3 )OH; ^R1 ' is selected from hydrogen, methyl, and ethyl; _A1 , _A2 , and _A3 are each independently selected from CH; X and Y are each independently selected from alkynyl groups; ^R2 is _-CH2OH ; m is 1. 4. The following compounds, their pharmaceutically acceptable salts, deuterated derivatives, or stereoisomers, are selected from: 5. A pharmaceutical composition comprising the compound of any one of claims 1-4, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, or a stereoisomer thereof. 6. Use of the compounds of claims 1-4, their pharmaceutically acceptable salts, deuterated derivatives or stereoisomers thereof, or the pharmaceutical composition of claim 5 in the preparation of a medicament for the treatment and/or prevention of infectious diseases. 7. The use as claimed in claim 6, wherein the infectious disease is an infectious disease caused by Gram-negative bacteria.