HK1090061B

HK1090061B - Novel lincomycin derivatives possessing antimicrobial activity

Info

Publication number: HK1090061B
Application number: HK06110528.4A
Authority: HK
Inventors: J．G．勒维斯; S．K．安纳丹; H．欧多德; M．F．格迪弗
Original assignee: 维库罗恩医药品公司
Priority date: 2003-06-17
Filing date: 2004-06-17
Publication date: 2010-09-30

Description

Lincomycin derivatives with antimicrobial activity

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part application of U.S. patent application No.10/777,455 filed on 11/2/2004, which is a continuation-in-part application of U.S. patent application No.10/642,807 filed on 15/8/2003, and further claims priority from 35u.s.c. § 119(e) U.S. provisional application No.60/479,296 filed on 17/6/2003 and U.S. provisional application No.60/749,502 filed on 17/6/2003, the disclosures of which are incorporated herein by reference in their entirety.

Background

Technical Field

The present invention relates to lincomycin derivatives that exhibit antibacterial activity and methods of using such derivatives.

State of the art

Lincomycin is a biosynthetic product that adversely affects the growth of a number of different microorganisms, particularly gram-positive bacteria. The characteristics and preparation of lincomycin are disclosed in U.S. Pat. No.3,086,912. Various lincomycin derivatives have been prepared that also possess antimicrobial activity. Such derivatives include, for example, clindamycin, as described in U.S. patent No.3,496,163.

Lincomycin derivatives remain attractive targets for antibacterial drug discovery. Thus, there is a need for lincomycin derivatives with antimicrobial activity as potential antibacterial agents.

Summary of The Invention

The present invention provides lincomycin derivatives having antibacterial activity. In some embodiments, the novel lincomycin derivatives exhibit antibacterial activity against gram-positive and anaerobic bacterial pathogens. Surprisingly, the selected novel lincomycin compounds described herein exhibit atypical efficacy against enterococcus species, such as enterococcus faecium and enterococcus faecalis (enterococcus faecium), and/or against fastidious gram-negative pathogens, such as haemophilus influenzae, compared to known compounds, such as clindamycin.

In one of its compositional aspects, the present invention relates to compounds of formula (I):

wherein:

w is a nitrogen-containing ring:

wherein m is 0, 1, 2 or 3; wherein when m is 2, the nitrogen-containing ring may optionally contain a double bond between the 4 and 5 nitrogen-containing ring positions; wherein when m is 3, the nitrogen containing ring may optionally contain a double bond between the 4 and 5 nitrogen containing ring positions or between the 5 and 6 nitrogen containing ring positions; wherein the nitrogen-containing ring positions are numbered consecutively in the counterclockwise direction starting with nitrogen as "1";

R¹selected from the group consisting of hydrogen, alkyl, substituted alkyl, hydroxyalkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cycloalkylalkyl, halo, alkylthio, and substituted alkylthio;

R²And R³Independently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cyano, alkylthio, substituted alkylthio, hydroxy, halo, or R²And R³One is NOR⁷The other is absent, or R²And R³One is ═ CH₂And the other is absent;

R⁶selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, substituted alkyl, aminomethyl, -C (O) O-alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, - (amido) alkyl, (carbamoyl) alkyl, 5-alkyl- [1, 3]Dioxolen-2-one-4-yl-methyl, 5-alkyl- [1, 3 [ ]]Dioxol-2-one-4-yl-methoxy-carbonyl group, or-N (R)⁶) -the fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure;

R⁷is H or alkyl;

R⁹may be mono-or multiply substituted on the same or different carbons in the ring, independently selected from hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkylalkyl, substituted alkenyl, substituted oxygen, substituted nitrogen, halogen, phenyl, substituted phenyl, alkylthio, substituted arylthio, heteroarylthioalkyl, heterocyclylthioalkyl, heteroarylthio, heterocyclylthio, propylene (═ CHCH) ₂CH₃) Azido, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵And branched isomers thereof (wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is H or alkyl), alkoxyalkoxy, aryl, substituted aryl, alkenyl, substituted alkenyl and-S (O)_qR¹³(wherein q is an integer equal to 0, 1 or 2, R¹³Selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl and substituted heterocyclyl), wherein there is no more than one-S (O) on the nitrogen-containing ring_qR¹³A group;

or a prodrug and/or a pharmaceutically acceptable salt thereof.

In another of its compositional aspects, the present invention relates to compounds of formula (II):

wherein:

w is a nitrogen-containing ring:

R²⁰And R²¹Independently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cyano, alkylthio, substituted alkylthio, hydroxy, halo, or R²⁰And R²¹One is NOR⁷The other is absent, or R²⁰And R²¹One is ═ CH₂The other is absent, or R²⁰And R²¹Together is cycloalkyl, aryl, substituted aryl, heterocyclyl or heteroaryl;

R⁶selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, substituted alkyl, aminomethyl, -C (O) O-alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, - (amido)Alkyl, (carbamoyl) alkyl, 5-alkyl- [1, 3%]Dioxolen-2-one-4-yl-methyl, 5-alkyl- [1, 3 [ ]]Dioxol-2-one-4-yl-methoxy-carbonyl group, or-N (R)⁶) -the fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure;

R⁷is H or alkyl;

or a prodrug and/or a pharmaceutically acceptable salt thereof.

In another of its compositional aspects, the present invention relates to compounds of formula (IA):

wherein:

represents a bond, which may be a double bond or a single bond;

R¹selected from the group consisting of-S-alkyl, -S-substituted alkyl, hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, and halo;

R²and R³Independently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cyano, alkylthio, substituted alkylthio, hydroxy, halo, or R²And R³One is NOR ⁷The other is absent, or R²And R³One is ═ CH₂And the other is absent;

R⁶selected from the group consisting of hydrogen, alkyl, substituted alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, - (amido) alkyl, (carbamoyl) alkyl, substituted aryl, substituted heteroaryl,

Group of or-N (R)⁶) -the fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure;

R⁷selected from the group consisting of hydrogen and alkyl;

R⁹may be mono-or multiply-substituted on the same or different carbons of the ring, and is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, aryl, substituted aryl, alkenyl, substitutedAlkenyl of and-S (O)_qR¹³Group of (I), wherein q is an integer equal to 0, 1 or 2, R¹³Selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, and substituted heterocyclyl;

wherein m is¹＝0-2；

Wherein t is 0-3;

or a pharmaceutically acceptable salt and/or prodrug thereof;

With the following conditions:

A. in the compounds of the formula (I), ifIs a single bond, and is,

m¹is a number of 0 or 1, and,

R²and R³Independently hydrogen, alkyl, hydroxy, fluoro, cyanoalkyl, or R²And R³One is NOR⁷The other is absent, or R²And R³One is ═ CH₂And the other is not present,

R⁶is hydrogen, alkyl, hydroxyalkyl, -C (O) O-alkylene-cycloalkyl, -C (O) O-alkylene-substituted alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, -C (O) O-heterocyclyl, -C (O) O-substituted heterocyclyl, - [ C (O) O ] O]_p-alkylene-heterocycle, - [ C (O) O]_p-alkylene-substituted heterocycle, wherein p is 0 or 1,

R⁷selected from the group consisting of hydrogen and alkyl,

R⁹is hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, haloPhenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is a hydrogen or an alkyl group,

then R is¹Is not-S-alkyl;

B. in the compounds of the formula (I), ifIs a single bond, and is,

R²And R³Independently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cyano, alkylthio, substituted alkylthio, hydroxy, halo, or R²And R³One is NOR⁷The other is absent, or R²And R³One is ═ CH₂And the other is absent, with the proviso that R is²And R³Not all are hydrogen; when R is²And R³When one is halo, the other is not hydrogen or hydroxy; when R is²And R³When one is a hydroxyl group, the other is not hydrogen or a hydroxyl group,

R⁶selected from the group consisting of hydrogen, alkyl, substituted alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, - (amido) alkyl, (carbamoyl) alkyl, or-N (R)⁶) The fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure,

R⁷selected from the group consisting of hydrogen and alkyl,

R¹selected from the group consisting of-S-alkyl, -S-substituted alkyl, hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, and halo,

then at least one R⁹Is not hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH) ₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is hydrogen or alkyl;

C. in the compounds of the formula (I), ifIs a single bond, and is,

R⁷selected from the group consisting of hydrogen and alkyl,

R¹selected from the group consisting of-S-alkyl, -S-substituted alkyl, (heteroaryl) alkyl, hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, and halo,

R⁹independently selected from the group consisting of other than (other than) hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH) ₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is a hydrogen or an alkyl group,

then R is⁶Selected from the group consisting of substituted alkyl (other than monosubstituted or substituted heterocycle),

(amido) alkyl and-N (R)⁶) -a group of fragments, -N (R)⁶) -the fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure;

wherein as used only in these conditions, the following specific terms have the following specific meanings:

substituted alkyl denotes alkyl in which one or more hydrogen atoms have been replaced by halogen, oxygen, hydroxyl, amine (primary), amine (secondary-alkyl substituted by the above-mentioned alkyl), amine (tertiary-alkyl substituted by alkyl as defined above), sulfur, -SH or phenyl,

substituted cycloalkyl denotes cycloalkyl substituted by alkyl, wherein alkyl is as defined above, or a group wherein one or more hydrogen atoms have been replaced by halogen, oxygen, hydroxy, amine (primary), amine (secondary-alkyl substituted by the above alkyl), amine (tertiary-alkyl substituted by alkyl as defined above), sulfur, -SH or phenyl,

substituted oxygen represents the group-OR ^dWherein R is^dAre alkyl, haloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alkenyl, cycloalkyl and substituted cycloalkyl,

the substituted nitrogen or amino group representing the radical-NR^aR^bWherein R is^aAnd R^bIndependently hydrogen, alkyl, haloalkyl, alkenyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl,

substituted aryl represents an aryl ring substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, alkoxy, acyloxy, amino, hydroxy, carboxy, cyano, nitro, alkylthio, and thioalkyl (thioalkyl), wherein alkylthio represents the group-S-alkyl, thioalkyl represents alkyl having one or more-SH groups,

substituted heteroaryl represents a heteroaryl ring substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, alkoxy, acyloxy, amino, hydroxy, carboxy, cyano, nitro, alkylthio, and thioalkyl, wherein alkylthio represents the group-S-alkyl and thioalkyl represents alkyl with one or more-SH groups.

In another of its compositional aspects, the present invention relates to compounds of formula (IB):

wherein:

R¹selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cycloalkylalkyl, halo, and substituted alkylthio;

R²and R³Independently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cyano, alkylthio, substituted alkylthio, hydroxy, halo, or R²And R³One is NOR⁷And the other is absent;

R⁶is H,Alkyl or hydroxyalkyl;

R⁷is H or alkyl;

R⁹may be mono-or multiply-substituted on the same or different carbons in the ring, and is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted oxygen, substituted nitrogen, halogen, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵And branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is H or alkyl;

m is 1 or 2;

or a prodrug and/or a pharmaceutically acceptable salt thereof.

In some embodiments, if the nitrogen-containing ring is saturated,

R²and R³Independently hydrogen, hydroxy, halo, alkoxy, alkylthio, substituted alkylthio, alkyl, substituted alkyl, hydroxyalkyl,

R⁶Is hydrogen, alkyl, hydroxyalkyl;

R⁹is hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵And branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is a hydrogen or an alkyl group,

then R is¹Is not-S-alkyl.

In some embodiments, if the nitrogen-containing ring is saturated,

m is 0, 1, 2 or 3,

R²and R³Independently hydrogen, alkyl, hydroxy, fluoro, cyanoalkyl, orR is²And R³One is NOR⁷The other is absent, or R²And R³One is ═ CH₂And the other is not present,

R⁷selected from the group consisting of hydrogen and alkyl;

R⁹is hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH) ₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is a hydrogen or an alkyl group,

then R is¹Is not-S-alkyl.

In some embodiments, if the nitrogen-containing ring is saturated,

m is a number of 1 or 2,

R⁶is hydrogen, alkyl, hydroxyalkyl, -C (O) O-alkylene-cycloalkyl, -C (O) O-alkylene-substituted alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) C-alkyl(O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, -C (O) O-heterocyclyl, -C (O) O-substituted heterocyclyl, - [ C (O) O ] O]_p-alkylene-heterocycle, - [ C (O) O]_p-alkylene-substituted heterocycle, wherein p is 0 or 1,

R⁷selected from the group consisting of hydrogen and alkyl,

R⁹is hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R ⁴And R⁵Is a hydrogen or an alkyl group,

then R is¹Is not-S-alkyl.

In some embodiments, if the nitrogen-containing ring is saturated,

R⁶selected from hydrogen, alkyl, substituted alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, - (amido) alkyl, (carbamoyl) alkyl, or-N (R)⁶) The fragment is an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkaneA part of the structure of an oxyamidine,

R⁷selected from the group consisting of hydrogen and alkyl;

Then at least one R⁹Is not hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is hydrogen or alkyl.

In some embodiments, if the nitrogen-containing ring is saturated,

R⁷selected from the group consisting of hydrogen and alkyl,

R⁹Independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is a hydrogen or an alkyl group,

(amido) alkyl and-N (R)⁶) -a group of fragments, -N (R)⁶) The fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure.

When used only in these conditions, the following specific terms have the following specific meanings:

Substituted oxygen represents the group-OR^dWherein R is^dIs alkyl, haloalkyl, arylSubstituted aryl, heteroaryl, substituted heteroaryl, alkenyl, cycloalkyl and substituted cycloalkyl groups,

substituted aryl represents an aryl ring substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, alkoxy, acyloxy, amino, hydroxy, carboxy, cyano, nitro, alkylthio, and thioalkyl, wherein alkylthio represents the group-S-alkyl, thioalkyl represents alkyl having one or more-SH groups,

In some embodiments, R²And R³Not all are hydrogen. In some embodiments, when R²And R³When one is halo, the other is not hydrogen or hydroxy. In some embodiments, when R²And R³When one is a hydroxyl group, the other is not hydrogen or a hydroxyl group.

In one embodiment, m is 0(W is). In another embodiment, m is 1 (W). In one embodiment, m is 2.

In another embodiment, m is 2 and the nitrogen containing ring is saturated (W is)。

In another embodiment, m is 2 and the nitrogen containing ring contains a double bond between the 4 and 5 nitrogen containing ring positions (W is). In one embodiment, m is 3. In that

In another embodiment, m is 3 and the nitrogen containing ring is saturated (W)). In that

In another embodiment, m is 3 and the nitrogen containing ring contains a double bond (W) between the 4 and 5 nitrogen containing ring positions). In another embodiment, m is 3 and the nitrogen containing ring contains a double bond between the 5 and 6 nitrogen containing ring positions (W is). In one embodiment, the nitrogen-containing ring is saturated.

In a preferred embodiment, the present invention provides compounds wherein the nitrogen-containing ring in the above formula is selected from

In one embodiment, R¹Selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, halo, alkylthio, and substituted alkylthio. In one embodiment, R ¹Selected from the group consisting of hydrogen, alkyl, substituted alkyl, hydroxyalkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cycloalkylalkyl, halo, and substituted alkylthio. In one embodiment, R¹Selected from the group consisting of hydrogen, alkyl, substituted alkyl, hydroxyalkyl, alkoxy, cycloalkylalkyl, alkylthio, and substituted alkylthio. In one embodiment, R¹Selected from the group consisting of hydrogen, alkyl, substituted alkyl, hydroxyalkyl, alkoxy, cycloalkylalkyl, and substituted alkylthio. In a preferred embodiment, R¹Selected from the group consisting of hydrogen, -S-methyl, -S-isopropyl, -S-tert-butyl, propyl, 2, 2, 2-trifluoro-ethyl-thio, 2-ethoxy-eth-1-yl, butoxy, 2-hydroxy-ethyl, 3-hydroxy-propyl, hydroxy-methyl, 2- (methyl-thio) -ethyl and cyclopropyl-methyl. In another preferred embodiment, R¹Selected from the group consisting of hydrogen, -S-isopropyl, -S-tert-butyl, propyl, 2, 2, 2-trifluoro-ethyl-thio, 2-ethoxy-eth-1-yl, butoxy, 2-hydroxy-ethyl, 3-hydroxy-propyl, hydroxy-methyl, 2- (methyl-thio) -ethyl and cyclopropyl-methyl. In another preferred embodiment, R ¹is-S-methyl. Preferred R¹Groups can be found in tables I, II and III. In some embodiments, R¹Is not-S-alkyl. In some embodiments, R¹Is not-S-methyl. In other embodiments, R¹Is not-S-substituted alkyl.

In other embodiments, R¹Is preferably-SR⁰Wherein R is⁰Preferably is C_1-4Alkyl is more preferably methyl, 2-hydroxyethyl or 2-ethylwaterAnd (3) a salicylate. In another embodiment, R¹Preferably hydrogen, alkyl, substituted alkyl or 2, 2, 2-trifluoroethylthio. More preferably, R¹Is hydrogen, propyl, 2-ethoxyethyl or 2, 2, 2-trifluoroethylthio.

In one embodiment, R²And R³Independently selected from the group consisting of hydrogen, alkyl, hydroxy, and halo. In a preferred embodiment, R²And R³Independently selected from the group consisting of hydrogen, methyl, hydroxy and chloro. In another preferred embodiment, R²And R³Are hydrogen and hydroxyl. In another preferred embodiment, R²And R³Are hydrogen and chloro. In another preferred embodiment, R²And R³Are hydrogen and methyl. Preferred R²And R³Groups can be found in tables I, II and III.

In one embodiment, R ²⁰And R²¹Independently is alkyl or alkenyl, or R²⁰And R²¹Together are cycloalkyl, aryl, substituted aryl, heterocyclyl or heteroaryl. In one embodiment, R²⁰And R²¹One is H and the other is alkyl or alkenyl. In a preferred embodiment, R²⁰And R²¹One is H and the other is ethyl or vinyl. In another embodiment, R²⁰And R²¹Together are cycloalkyl or aryl. In a preferred embodiment, R²⁰And R²¹Together is cyclopropyl, cyclopentyl, phenyl or 4-chloro-phenyl. Preferred R²⁰And R²¹Groups can be found in tables I, II and III. In one embodiment, if R²⁰And R²¹One is hydrogen and the other is not hydrogen, alkyl, hydroxy, cyano, alkylthio or substituted alkylthio.

In one embodiment, R⁶Selected from the group consisting of hydrogen, alkyl, cycloalkyl, hydroxyalkyl, substituted alkyl, aminomethyl, -C (O) O-substituted alkyl, 5-alkyl- [1, 3]Dioxolen-2-one-4-yl-methyl and 5-alkyl-[1，3]Dioxol-2-one-4-yl-methoxy-carbonyl. In another embodiment, R⁶Selected from hydrogen and alkyl. In one embodiment, R⁶Selected from the group consisting of 1H-imidazol-2-yl-methyl, 2- [ HC (O) ]-eth-1-yl, 2-amino-eth-1-yl, 2-hydroxyethyl, 2-methoxy-eth-1-yl, 5-methyl-2-oxo- [1, 3]Dioxolen-4-yl-methoxy-carbonyl, 5-methyl-2-oxo- [1, 3]Dioxol-4-yl-methyl, aminocarbonylmethyl, aminocarbonylethyl, cyanomethyl, cyclopropyl, hydrogen, iminomethyl, methyl and methoxycarbonylmethyl. In one embodiment, R⁶Selected from the group consisting of 1H-imidazol-2-yl-methyl, 2-hydroxyethyl, 5-methyl-2-oxo- [1, 3]Dioxolen-4-yl-methoxy-carbonyl, 5-methyl-2-oxo- [1, 3]Dioxol-4-yl-methyl, aminocarbonylmethyl, cyanomethyl, cyclopropyl, hydrogen, iminomethyl and methyl. In a preferred embodiment, R⁶Selected from the group consisting of 1H-imidazol-2-yl-methyl, 2- [ HC (O)]-eth-1-yl, 2-amino-eth-1-yl, 2-hydroxyethyl, 2-methoxy-eth-1-yl, aminocarbonylmethyl, aminocarbonylethyl, cyanomethyl, cyclopropyl, hydrogen, iminomethyl, methyl and methoxycarbonylmethyl. In a preferred embodiment, R⁶Is hydrogen or methyl. In another preferred embodiment, R⁶Selected from the group consisting of 5-methyl- [1, 3]Dioxolen-2-one-4-yl-methyl and 5-methyl- [1, 3 [ ] ]Dioxol-2-one-4-yl-methoxy-carbonyl. Preferred R⁶Groups can be found in tables I, II and III.

In another embodiment, R⁹Selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, cycloalkylalkyl, substituted alkenyl, alkylthio, substituted arylthio, heteroarylthioalkyl, heterocyclylthioalkyl, halogen, and propylene (═ CHCH)₂CH₃) Azido, substituted oxygen, heteroarylthio and heterocyclylthio. In another embodiment, R⁹Selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, cycloalkylalkyl, substituted alkenyl, alkylthio, substituted arylthioHeteroarylthioalkyl, heterocyclylthioalkyl, halogen, propylene (═ CHCH)₂CH₃) And an azide group. In a preferred embodiment, R⁹Is an alkyl group. In another preferred embodiment, R⁹Is a halogen.

In another embodiment, R⁹Selected from the group consisting of (2-fluorocyclopropyl) methoxy, (3-fluoropropoxy) methyl, 1H-pyrrolylmethyl, 2- (4-ethylthiazol-2-yl) -eth-1-yl, 2- (4-methylthiazol-2-yl) -eth-1-yl, 2- (5-ethyl-isoxazol-3-yl) -eth-1-yl, 2, 2, 2-trifluoroethyl-thio, 2, 2-difluoroethoxymethyl, 2- [1, 3 ] methyl ]Dithiolan-2-yl-eth-1-yl, 2-chlorophenyl-methylthio, 2-cyclobutylethyl, 2-cyclopropylethyl, 2-mercaptoethoxy-ethyl-thio, 2-fluoroethoxy, 2-propoxyethyl, 3- (1H- [1, 2, 3)]Triazole) -propan-1-yl, 3- (3-fluoropropoxy) propyl, 3- (cyclohexyloxy) propyl, 3- (difluoromethylthio) propyl, 3- (ethylthio) propyl, 3- (furan-2-ylmethylthio) -propan-1-yl, 3, 3, 3-trifluoropropan-1-yl-thio, 3, 3, 3-trifluoropropoxy, 3, 3-difluoroallyl, 3, 3-difluorobutyl, 3, 3-difluoropropyl, 3- [ (cyclopropyl) methoxy-propyl]Propyl, 3-cyanoprop-1-yl, 3-cyclohexyloxypropyl, 3-cyclopropyl-propyl, 3-ethoxyiminoprop-1-yl, 3-ethylthioprop-1-yl, 3-fluoropropoxy, 3-fluoropropoxymethyl, 3-fluoropropyl, 3-imidazol-1-yl-prop-1-yl, 3-mercaptopropylthio, 3-methoxyimino-prop-1-yl, 3-methylbut-1-ylthio, 3-methylbutyl, 3-pyridin-4-yl-allyl, 3-pyridin-4-yl-propyl, 3-pyrrolidin-2-keto-prop-1-yl, 3-ethoxyiminoprop-1-yl, 3-thiopropyl-1-yl, 3-methylpropyloxy, 3-pyridin-4-yl-allyl, 3-pyridin-1-yl, 3-thiophen-2-ylthiopropan-1-yl, 4- (methoxy) butyl, 4-difluorobutyl, 4-difluoropentyl, 4-fluorobutoxy, 5-difluoropentyl, azido, butoxy, butyl, butylthio, chloro, cyclobutylmethyl, cyclohexylmethyl, cyclopropyl, cyclopropylmethyl, ethyl, ethylthio, fluoro, isobutyl, methyl, m-methylbenzylthio, n-butylthio, o, p-dichlorobenzylthio, pentyloxy, pentyl, p-fluorobenzylthio, p-methylbenzylthio; propoxy, propyl, and propylene (═ CHCH) ₂CH₃)、P-trifluoromethoxy benzylthio, pyrazin-2-yl-methyl-thio, pyridin-4-yl-thio and thiophen-2-yl-methylthio.

In another embodiment, R⁹Selected from 2- (4-methylthiazol-2-yl) -ethan-1-yl, 2- (5-ethyl-isoxazol-3-yl) -ethan-1-yl, 2- [1, 3]Dithiolan-2-yl-eth-1-yl, 2-cyclobutylethyl, 2-cyclopropyl-ethyl, 3- (difluoromethylthio) -prop-1-yl, 3- (furan-2-ylmethylthio) -prop-1-yl, 3, 3, 3-trifluoroprop-1-yl-thio, 3, 3-difluoroallyl, 3, 3-difluoro-propyl, 3-cyanoprop-1-yl, 3-cyclopropyl-propyl, 3-ethoxyiminoprop-1-yl, 3-ethylthioprop-1-yl, 3-imidazol-1-yl-prop-1-yl, di-thiocyano-prop-1-yl, di-azolo-prop-1-yl, di-ethylthio-prop-1, 3-methoxyimino-prop-1-yl, 3-methylbut-1-yl-thio, 3-methylbutyl, 3-pyridin-4-yl-allyl, 3-pyridin-4-yl-propyl, 3-thiophen-2-ylthioprop-1-yl, 4-propyl, azido, butyl, butylthio, cyclobutylmethyl, cyclopropyl, cyclopropylmethyl, ethyl, ethylthio, fluoro, methyl, n-butylthio, o, p-dichlorobenzylthio, pentyl, p-fluorophenylthio, p-methylbenzylthio, propyl, propylene (═ CHCH- ₂CH₃) Pyrazin-2-yl-methyl-thio and thiophen-2-yl-methylthio. In one embodiment, at least one R is⁹The radical is not hydrogen.

In a preferred embodiment, R⁹Is propyl. Preferred R⁹Groups can be found in tables I, II and III.

In one embodiment, Z is selected from the group consisting of hydrogen, phosphate, and palmitate. In one embodiment, Z is hydrogen. In another embodiment, Z is a phosphate. In another embodiment, Z is palmitate.

The compounds of the present invention also include prodrugs of formula (I), (II), (IA) and (IB). Such prodrugs include compounds of formulae (I), (II), (IA) and (IB) wherein R⁶Or one of the hydroxyl groups on the sugar is modified to include a group selected from phosphate, palmitate or

A substituent of (1).

Preferred prodrugs include compounds of formulae (I), (II), (IA) and (IB) wherein R⁶Or one of the hydroxyl groups on the sugar is modified to include a group selected from

A substituent of (1).

Preferred compounds of formulae (I), (II), (IA) and (IB) have a minimum inhibitory concentration of 32 μ g/ml or less against at least one organism selected from the group consisting of: streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, enterococcus faecalis, enterococcus faecium, Haemophilus influenzae, Moraxella catarrhalis, Escherichia coli, Bacteroides fragilis, Bacteroides thetaiotaomicron, and Clostridium difficile.

In one embodiment, the compounds of formulae (I), (II), (IA) and (IB) have a minimum inhibitory concentration of 4 μ g/ml or less against at least one organism selected from the group consisting of: haemophilus influenzae and moraxella catarrhalis. In one embodiment, the compounds of formulae (I), (II), (IA) and (IB) have a minimum inhibitory concentration of 4 μ g/ml or less against at least one organism selected from the group consisting of: enterococcus faecalis and enterococcus faecium. In one embodiment, the compounds of formulae (I), (II), (IA) and (IB) have a minimum inhibitory concentration of 4 μ g/ml or less against at least one organism selected from the group consisting of: the gram-negative organisms haemophilus influenzae VHIN1003 and haemophilus influenzae VHIN 1004.

Another aspect of the invention is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound described herein.

Another aspect of the invention is a method of treating a microbial infection in a mammal comprising administering to the mammal a therapeutically effective amount of a compound described herein. In one embodiment, the microbial infection being treated is caused by one or more of the following pathogens: haemophilus influenzae, moraxella catarrhalis, enterococcus faecalis, and enterococcus faecium. The administered compound may be formulated into a pharmaceutical composition as described herein. The compounds may be administered to a mammal in a pharmaceutical composition orally, parenterally, transdermally, topically, rectally, or intranasally. In one embodiment, the compound may be administered in an amount of from about 0.1 to about 100mg/kg body weight/day.

Lincomycin within the scope of the present invention includes those of formula I as depicted in Table I below, wherein the nitrogen-containing ring positions are numbered consecutively in a counter-clockwise direction starting with the nitrogen as "1", i.e.

TABLE I

In Table I, R is when m is 0 or 1, unless otherwise noted⁹The substituent is substituted at the 4-position.

Lincomycin derivatives that are also within the scope of the present invention include those of formula II as described in table II below, wherein the nitrogen-containing ring positions are numbered as in formula (I).

TABLE II

Lincomycin derivatives that are also within the scope of the present invention include those of formula III as described in table III below:

wherein the nitrogen containing ring position is numbered as in formula (I).

TABLE III

Ex.No.	R	Z	R/R	R	R	m
Ex.No.	R	Z	R/R	R	R	m	115	Propyl radical	-P(＝O)(OH)	H/Me	H	4-pentyl radical	1
116	Propyl radical	-C(O)(CH)-CH	H/Me	H	4-pentyl radical	1	115	Propyl radical	-P(＝O)(OH)	H/Me	H	4-pentyl radical	1
116	Propyl radical	-C(O)(CH)-CH	H/Me	H	4-pentyl radical	1	117	Propyl radical	-P(＝O)(OH)	H/Me	H	4-propyl radical	1
118	Propyl radical	-C(O)(CH)-CH	H/Me	H	4-propyl radical	1	117	Propyl radical	-P(＝O)(OH)	H/Me	H	4-propyl radical	1

In the above tables I, II and III, the following abbreviations are used:

s-single bond

D ═ double bond

D4, 5 ═ double bonds between the 4 and 5 nitrogen-containing ring positions

Me is methyl

Pr ═ propyl radical

Bu ═ butyl

i ═ iso-

t ═ tert-

As used hereinafter, these compounds are named based on amine derivatives, but alternatively, these compounds may also be named based on 1-thio-L-threo- α -D-galacto-pyranoside derivatives.

Preferred compounds within the scope of the present invention include the following:

4- (3, 3-difluoro-allyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-pyridin-4-yl-allyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-pyridin-4-yl-propyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-butylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-ethylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-ethylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1-1 (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-ethylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (4-methyl-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (4-fluoro-phenylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3, 3, 3-trifluoro-propylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-methyl-butylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (2, 4-dichloro-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (thiophen-2-ylmethylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (pyrazin-2-ylmethylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (2, 4-dichloro-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-butylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-azido-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [3- (furan-2-ylmethylsulfanyl) -propan-1-yl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-imidazol-1-yl-propan-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [3- (thiophen-2-ylsulfanyl) -propan-1-yl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-ethylsulfanyl-prop-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-cyano-prop-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-difluoromethylsulfanyl-propan-1-yl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-difluoromethylsulfanyl-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (2- [1, 3] dithiolan-2-yl-ethyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [2- (4-methyl-thiazol-2-yl) -ethyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-methoxyimino-propan-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-ethoxyimino-propan-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [2- (5-ethyl-isoxazol-3-yl) -ethyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6 methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-propyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-butyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-ethyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propylidene-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-carbamoylmethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-cyanomethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1- (1H-imidazol-2-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-aminomethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-isopropylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-isopropylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [1- (6-tert-butylsulfanyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [1- (6-tert-butylsulfanyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -5-propyl-azepan-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -5-propyl-azepan-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester;

5-methyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-ethyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-cyclopropylmethyl-azepan-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-cyclopropyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-ethyl-4-methyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-ethyl-5-methyl-azepan-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-ethyl-6-methyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-fluoro-5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-butyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-1- (2-hydroxy-ethyl) -4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-butyl-4-fluoro-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (2-cyclobutyl-ethyl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-cyclopropylmethyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-cyclopropylmethyl-piperidine-2-carboxylic acid (2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (2-cyclobutyl-ethyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (2-cyclobutyl-ethyl) -pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-cyclobutylmethyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-propyl-2, 3, 6, 7-tetrahydro-1H-aza-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]-an amide;

4- (2-cyclopropyl-ethyl) -pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-cyclopropylmethyl-4-fluoro-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-cyclopropyl-5-propyl-azepane-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-butyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-propyl-azepane-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-isopropylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-propyl-azepane-2-carboxylic acid [1- (6-tert-butylsulfanyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

3-cyclopropylmethyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3- (2-cyclobutyl-ethyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3- (2-cyclopropyl-ethyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3- (3-cyclopropyl-propyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3-propyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3-butyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3-butyl-1- (2-hydroxy-ethyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3-pentyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3- (3-methyl-butyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3- (3, 3-difluoro-propyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

3-butyl-1-methyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ cyclopropyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [ cyclopropyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

5-propyl-azepane-2-carboxylic acid [ cyclopropyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [ phenyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ phenyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [ cyclopentyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ cyclopentyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

5-propyl-azepane-2-carboxylic acid [ cyclopentyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

5-propyl-azepane-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -butyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -but-3-enyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -but-3-enyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -butyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ (4-chloro-phenyl) - (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [ (4-chloro-phenyl) - (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -butyl ] -amide;

5-propyl-azepane-2-carboxylic acid [ (4-chloro-phenyl) - (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide;

4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-piperidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (2, 2, 2-trifluoro-ethylsulfanyl) -tetrahydro-pyran-2-yl ] -propyl } -amide;

4-pentyl-pyrrolidine-2-carboxylic acid [1- (6-ethoxyethyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

1- (2-hydroxy-ethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-pentyl-pyrrolidine-2-carboxylic acid [1- (6-butoxy-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

4-butyl-1-methyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

mono- (4, 5-dihydroxy-6- { 2-methyl-1- [ (4-pentyl-pyrrolidine-2-carbonyl) -amino ] -propyl } -2-propyl-tetrahydro-pyran-3-yl) -phosphate;

hexadecanoic acid 4, 5-dihydroxy-6- { 2-methyl-1- [ (4-pentyl-pyrrolidine-2-carbonyl) -amino ] -propyl } -2-propyl-tetrahydro-pyran-3-yl ester;

mono- (4, 5-dihydroxy-6- { 2-methyl-1- [ (4-propyl-pyrrolidine-2-carbonyl) -amino ] -propyl-2-propyl-tetrahydro-pyran-3-yl) -phosphate;

hexadecanoic acid 4, 5-dihydroxy-6- { 2-methyl-1- [ (4-propyl-pyrrolidine-2-carbonyl) -amino ] -propyl } -2-propyl-tetrahydro-pyran-3-yl ester;

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-pentyl-pyrrolidine-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester;

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-propyl-pyrrolidine-2-carboxylic acid (2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-propyl-pyrrolidine-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester;

4-propyl-pyrrolidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (2-hydroxy-ethyl) -tetrahydro-pyran-2-yl ] -propyl } -amide;

4-propyl-pyrrolidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (3-hydroxy-propyl) -tetrahydro-pyran-2-yl ] -propyl } -amide;

4-propyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-hydroxymethyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-propyl-pyrrolidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (2-methylsulfanyl-ethyl) -tetrahydro-pyran-2-yl ] -propyl } -amide;

4-propyl-pyrrolidine-2-carboxylic acid [1- (6-cyclopropylmethyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

or a prodrug and/or a pharmaceutically acceptable salt thereof.

Further compounds within the scope of the invention include:

4- (4-fluoro-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (pyridin-2-ylmethylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1- (2-methoxy-ethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1- (2-formylamino-ethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1- (2-amino-ethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-cyclohexyloxy-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

{2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-pentyl-pyrrolidin-1-yl } -acetic acid methyl ester;

1-methylcarbamoylmethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [3- (furan-2-ylmethylsulfanyl) -propyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-imidazol-1-yl-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [3- (thiophen-2-ylsulfanyl) -propyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [3- (2-oxo-pyrrolidin-1-yl) -propyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-methoxyimino-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- [2- (4-ethyl-thiazol-2-yl) -ethyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-ethylsulfanyl-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-ethoxyimino-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-pyrrol-1-ylmethyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-propyl-piperidine-1-carboxylic acid 9H-fluoren-9-ylmethyl ester;

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-propyl-piperidine-1-carboxylic acid ethyl ester;

4- (3-cyano-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-propyl-piperidine-1-carboxylic acid phenyl ester;

4- (2- [1, 2, 3] triazol-1-yl-ethyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-propyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-propyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4- (3-difluoromethylsulfanyl-propyl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

4-pentyl-pyrrolidine-2-carboxylic acid [1- (6-ethoxymethyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide;

or a prodrug and/or a pharmaceutically acceptable salt thereof.

Additional compounds of the invention include:

phosphoric acid mono- (6- { 2-chloro-1- [ (5-propyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester;

phosphoric acid mono- (6- { 2-chloro-1- [ (5-fluoro-5-propyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester;

phosphoric acid mono- (6- { 2-chloro-1- [ (5-cyclopropylmethyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester;

phosphoric acid mono- (6- { 2-chloro-1- [ (4-fluoro-4-propyl-piperidine-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester;

hexadecanoic acid 6- { 2-chloro-1- [ (5-propyl-azepane-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester;

hexadecanoic acid 6- { 2-chloro-1- [ (5-fluoro-5-propyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester;

Hexadecanoic acid 6- { 2-chloro-1- [ (5-cyclopropylmethyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester; and

hexadecanoic acid 6- { 2-chloro-1- [ (4-fluoro-4-propyl-piperidine-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester.

Additional compounds of the invention include:

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -5-fluoro-5-propyl-azepan-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester;

5-fluoro-1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

5-cyclopropylmethyl-1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide;

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -5-cyclopropylmethyl-azepan-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester;

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-fluoro-4-propyl-piperidine-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester; and

4-fluoro-1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-propyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

The compounds, prodrugs and pharmaceutically acceptable salts thereof, as defined herein, may have activity against bacteria, protozoa, fungi and/or parasites.

In another aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound as defined herein. The pharmaceutical compositions of the present invention may further comprise one or more additional antibacterial agents. In one embodiment, the one or more additional antimicrobial agents may be active against gram-negative bacteria. In one embodiment, the one or more additional antibacterial agents may be active against gram positive bacteria. In another embodiment, the at least one antimicrobial agent may be active against both gram negative and gram positive bacteria.

In one of its method aspects, the present invention relates to a method of treating a microbial infection in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of the present invention. The compounds of the present invention may be administered to a mammal in a pharmaceutical composition orally, parenterally, transdermally, topically, rectally, or intranasally.

In another of its method aspects, the present invention relates to a method of treating a microbial infection in a mammal comprising administering to the mammal a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention. The pharmaceutical compositions of the present invention may further comprise one or more additional antibacterial agents. In one embodiment, the one or more additional antimicrobial agents may be active against gram-negative bacteria. In one embodiment, the one or more additional antibacterial agents may be active against gram positive bacteria. The pharmaceutical compositions can be administered to a mammal orally, parenterally, transdermally, topically, rectally, or intranasally.

In a preferred embodiment, the microbial infection treated is a gram positive infection. In another embodiment, the infection may be a gram negative infection. In further embodiments, the infection may be a mycobacterial infection, a mycoplasma infection, or a chlamydia infection.

In another aspect, the present invention provides novel intermediates and methods for preparing the compounds described herein.

Detailed description of the invention

As mentioned above, the present invention relates to lincomycin derivatives which exhibit antibacterial activity, in particular gram-positive antibacterial activity. In some embodiments, the novel lincomycin derivatives exhibit antibacterial activity against gram-positive and anaerobic bacterial pathogens. Surprisingly, the selected novel lincomycin compounds described herein exhibit atypical efficacy against enterococci, such as enterococcus faecium and enterococcus faecalis, and/or against fastidious gram-negative pathogens, such as haemophilus influenzae, compared to known compounds, such as clindamycin. However, before describing the present invention in further detail, the following terms will first be defined.

It must be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a "pharmaceutically acceptable carrier" includes a plurality of such carriers; reference to "an additional antimicrobial agent" is a reference to one or more drugs and equivalents thereof known to those skilled in the art, and so forth.

Definition of

Unless otherwise defined, the following terms used in the specification and claims have the meanings given below:

"acyl" represents the group-C (O) R¹⁴Wherein R is¹⁴Is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl or substituted heterocyclyl.

"acylamino" denotes-NR^aC(O)R¹⁴Wherein R is^aAnd R¹⁴Is as defined above.

"alkenyl" means a straight chain unsaturated monovalent hydrocarbon group of two to eight carbon atoms or a branched monovalent hydrocarbon group of three to eight carbon atoms containing at least one double bond (-C ═ C-), preferably 1-2 double bonds. Examples of alkenyl groups include, but are not limited to, allyl, vinyl, 2-butenyl, and the like.

"alkoxy" represents the group "alkyl-O-", and includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1, 2-dimethylbutoxy, and the like.

"alkyl" means a straight chain saturated monovalent hydrocarbon group of one to eight carbon atoms or a branched saturated monovalent hydrocarbon group of three to eight carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and the like.

"alkylene" means a straight chain divalent hydrocarbon radical of one to eight carbon atoms or a branched divalent hydrocarbon radical of three to eight carbon atoms. Examples of alkylene groups include, but are not limited to, methylene, ethylene, 2-methylpropylene, and the like.

"alkylthio" means a group "alkyl-S-", wherein alkyl is as defined herein, including, for example, methylthio, butylthio, and the like.

"alkynyl" means a straight-chain monovalent hydrocarbon radical of two to eight carbon atoms or a branched-chain monovalent hydrocarbon radical of three to eight carbon atoms, containing at least one triple bond (-C.ident.C-), preferably a single triple bond. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 2-butynyl, and the like.

"amino" or "substituted nitrogen" denotes the group "-NR^aR^b", wherein R^aAnd R^bIndependently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, or wherein R is^aAnd R^bTogether with the nitrogen atom to which they are bonded, form a heterocyclic ring.

"aminoacyl" means-C (O) NR^aR^b。

"Aminocarbonylalkyl" denotes the radical "-R^cC(O)NR^aR^b", wherein R^cIs alkylene, R^aAnd R^bIs as defined above.

"aryl" means a monovalent monocyclic or bicyclic aromatic carbocyclic group of 6 to 14 ring atoms. Examples include, but are not limited to, phenyl, naphthyl, and anthracenyl. The aryl ring may optionally be fused to a 5-, 6-or 7-membered monocyclic non-aromatic ring, which optionally contains 1 or 2 heteroatoms independently selected from oxygen, nitrogen or sulfur, the remaining ring atoms being C, wherein one or two C atoms are optionally replaced by a carbonyl group. Representative aryl groups with fused rings include, but are not limited to, 2, 5-dihydrobenzo [ b ]]Oxygen oxide2, 3-dihydrobenzo [1, 4 ]]Dioxane, chroman, isochroman, 2, 3-dihydrobenzofuran, 1, 3-dihydroisobenzofuran, benzo [1, 3 ]]Dioxoles, 1, 2, 3, 4-tetrahydroisoquinolines, 1, 2, 3, 4-tetrahydroquinolines, 2, 3-dihydro-1H-indoles, 2, 3-dihydro-1H-isoindoles, benzimidazol-2-ones, 2H-benzoxazol-2-ones, and the like.

"carboxy" represents the group "C (O) OH".

"cyanoalkyl" means an alkyl group substituted with one or more cyano groups (-CN), provided that only a single cyano group is present on the same carbon atom. Examples of cyanoalkyl groups include, for example, cyanomethyl, 2-cyanoethyl, 2-cyanopropyl and the like.

"cycloalkyl" means a cyclic saturated hydrocarbon group of 3 to 8 ring atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

"cycloalkylalkyl" denotes the radical-R^cR^dWherein R is^cIs alkylene, R^dAre cycloalkyl, all as defined above. Examples include, but are not limited to, cyclopropylmethylene, cyclohexylethylene, and the like.

"halo" or "halogen" means fluoro, chloro, bromo, or iodo.

"haloalkyl" means an alkyl group substituted with one or more, preferably one to six, identical or different halogen atoms. Examples of haloalkyl groups include, for example, trifluoromethyl, 3-fluoropropyl, 2-dichloroethyl, and the like.

"heteroaryl" means a monovalent monocyclic or bicyclic aromatic group of 5 to 10 ring atoms containing 1, 2 or 3 ring heteroatoms selected from N, O or S, the remaining ring atoms being C.

"heterocycle" or "heterocyclyl" means a saturated or unsaturated group having a single ring or multiple fused rings with 1 to 1 in the ring10 carbon atoms and 1 to 4 atoms selected from nitrogen, oxygen or S (O)_q(wherein q is 0, 1 or 2), wherein in the fused ring system, one or more of the rings may be aryl or heteroaryl.

Examples of heterocycles and heteroaryls include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indoline, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine; imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1, 2, 3, 4-tetrahydroisoquinoline, 4, 5, 6, 7-tetrahydrobenzo [ b ] thiophene, thiazole, thiazolidine, thiophene, benzo [ b ] thiophene, morpholinyl, thiomorpholinyl (also referred to as thiomorpholinyl), piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.

"hydroxy" represents the group-OH.

"hydroxyalkyl" means an alkyl group substituted with one or more-OH groups, provided that only a single hydroxyl group (-OH) is present on the same carbon atom. Examples of hydroxyalkyl groups include, for example, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, and the like.

"mammal" means all mammals, including humans, domestic animals and pets.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "aryl optionally mono-or di-substituted with alkyl" means that alkyl may, but need not, be present, and the description includes situations where aryl is mono-or di-substituted with alkyl and situations where aryl is not substituted with alkyl.

By "pharmaceutically acceptable carrier" is meant a carrier which is useful in preparing pharmaceutical compositions, which is generally safe, non-toxic, neither biologically nor otherwise undesirable, and which includes carriers which are acceptable for veterinary use as well as human pharmaceutical use. The term "pharmaceutically acceptable carrier" as used in the specification and claims includes one or more such carriers.

By "pharmaceutically acceptable salt" of a compound is meant a salt which is pharmaceutically acceptable and possesses the desired pharmacological activity of the parent compound. Such salts include:

(1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids, such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4' -methylenebis (3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, laurylsulfuric acid, Gluconic acid, glutamic acid, naphthoic acid, salicylic acid, stearic acid, mucic acid, and the like; or

(2) Salts formed when an acidic proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion, or is complexed with an organic base, such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, and the like.

By "prodrug" is meant any compound that, when administered to a mammalian subject, releases the active parent drug of a compound according to the invention in vivo. Prodrugs of the compounds of the present invention are prepared by modifying functional groups present in the compounds of the present invention in such a way that the modifications are cleaved, in vivo, to release the parent compound. Prodrugs include compounds of the present invention wherein a hydroxy, mercapto or amino group in the compound is bonded to any group that can be cleaved in vivo to regenerate the free hydroxy, mercapto or amino group, respectively. Examples of prodrugs include, but are not limited to, esters of hydroxy functional groups in the compounds of the present invention (e.g., acetate, formate, palmitate and benzoate derivatives), carbamates (e.g., N-dimethylaminocarbonyl), and the like. Preferred prodrug substituents include the following substituents attached to the N-position of a five or six membered nitrogen containing heterocyclic ring: phosphoric acid esters, palmitic acid esters or

"substituted alkyl" means an alkyl group as defined above having 1 to 3 substituents independently selected from the group consisting of cyano, halogen (i.e., Cl, Br, F OR I), acyl, substituted oxygen, hydroxy, alkylthio, substituted alkylthio, cycloalkyl, substituted cycloalkyl, aminocarbonylalkyl, carboxy, -C (O) H, -C (O) OR ¹⁵(wherein R is¹⁵Is alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, etc.), -C (O) NR^aR^bSubstituted nitrogen, ═ N-OR⁷(wherein R is⁷Is hydrogen or alkyl), -SH, -S (O)_qR¹⁶(wherein q is 0, 1 or 2, R¹⁶Is the group consisting of alkyl, haloalkyl, aryl, heteroaryl, heterocyclyl and alkyl substituted with aryl, heteroaryl, cycloalkyl and heterocyclyl), aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl and substituted heterocyclyl. Examples of substituted alkyl groups include, but are not limited to, 1-fluoroethyl, 1-chloroethyl, 2-fluoroethyl, 2-chloroethyl, 1-bromopropyl, 2-iodopropyl, 1-chlorobutyl, 4-fluorobutyl, 4-chlorobutyl, 2-ethoxyeth-1-yl, -CH₂-S(O)₂-CH₃And the like.

"substituted alkenyl" means alkenyl as defined above in which one or more hydrogen atoms, preferably 1 to 3 hydrogen atoms, have been replaced by a substituent as defined for substituted alkyl.

"substituted alkynyl" means an alkynyl group as defined above in which one or more hydrogen atoms, preferably 1 to 3 hydrogen atoms, have been replaced by a substituent as defined for substituted alkyl.

"substituted alkylthio" means a group-S-substituted alkyl wherein the substituted alkyl is as defined above, including, for example, 2-hydroxyethylthio and the like.

"substituted alkoxy" means a group-O-substituted alkyl, wherein substituted alkyl is as defined above.

"substituted aryl" means an aryl ring substituted with one OR more substituents, preferably one to three substituents, selected from the group consisting of alkyl, substituted alkyl, alkylthio, substituted alkylthio, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, halo, alkoxy, substituted alkoxy, acyl, amino, acylamino, cycloalkyl, substituted cycloalkyl, cycloalkylalkyl, heteroaryl, substituted heteroaryl, heterocyclyl, substituted heterocyclyl, hydroxy, carboxy, -C (O) OR¹⁵、-C(O)NR^aR^bCyano, nitro and sulfanylalkyl (sulfanylalkyl). The aryl ring may optionally be fused to a 5-, 6-or 7-membered monocyclic non-aromatic ring, which optionally contains 1 or 2 heteroatoms independently selected from oxygen, nitrogen or sulfur, the remaining ring atoms being C, wherein one or two C atoms are optionally replaced by a carbonyl group.

"substituted cycloalkyl" means cycloalkyl substituted with alkyl or a group as defined above for substituted alkyl. Representative examples include, but are not limited to, 2-cyclopropylethyl, 3-cyclobutylpropyl, 4-cyclopentylbutyl, 4-cyclohexylbutyl, and the like.

"substituted heteroaryl" means a heteroaryl ring substituted with one or more substituents, preferably one to three substituents, selected from the group as defined above for substituted aryl.

"substituted heterocyclyl" means a heterocyclic group that is independently substituted with 1 to 3 substituents as defined for substituted cycloalkyl.

"substituted oxygen" denotes the group "-O-R^d", wherein R^dIs alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl or substituted heterocyclyl.

"substituted phenyl" means a phenyl group having 1 to 3 substituents selected from the group defined for substituted aryl.

"thioalkylalkyl" means an alkyl group substituted with one or more-SH groups, provided that if two mercapto groups are present, they are not both on the same carbon atom. Examples of thioalkyl groups include, for example, thioalkyl methyl, 2-thioalkyl ethyl, 2-thioalkyl propyl, and the like.

By "therapeutically effective amount" is meant an amount of a compound that, when administered to a mammal to treat a disease, is sufficient to effect such treatment for the disease. The "therapeutically effective amount" will vary with the compound, the disease and its severity, the age, weight, etc., of the mammal being treated.

"treating" a disease or "treatment" of a disease includes:

(1) prevention of disease, i.e., causing clinical symptoms of disease not to occur in a mammal that may be exposed to the disease or predisposed to the disease but does not yet experience or display symptoms of the disease,

(2) inhibiting the disease, i.e. preventing or reducing the development of the disease or its clinical symptoms, or

(3) Remission of the disease, i.e., causing regression of the disease or its clinical symptoms.

The compounds of the invention are generally named according to the IUPAC or CAS nomenclature system. Abbreviations well known to those of ordinary skill in the art may be used (e.g., "Ph" for phenyl, "Me" for methyl, "Et" for ethyl, "h" for hours, and "rt" for room temperature).

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Before the present compositions and methods are described, it is to be understood that this invention is not limited to the particular methodology, protocols, assays, and reagents described, as such may, of course, vary. It is also to be understood that the terminology used herein is intended to describe particular embodiments of the invention, and is in no way intended to limit the scope of the invention as described in the appended claims.

Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are now described. All publications cited herein are incorporated herein by reference in their entirety for the purpose of describing and disclosing the methodologies, reagents, and tools reported in the publications that might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

The practice of the present invention will employ, unless otherwise indicated, conventional methods of chemistry, biochemistry, molecular biology, cell biology, and pharmacology, which are within the skill of the art. Such techniques are explained fully in the literature.

General synthetic procedure

The compounds of the invention can be prepared by the methods described in the reaction schemes shown below.

The starting materials and Reagents for preparing these Compounds are available from commercial suppliers, such as Torant Research Chemicals (North York, ON Canada), Aldrich chemical Co. (Milwaukee, Wisconsin, USA), Bachem (Torrance, California, USA), Emka-Chemistry or Sigma (St. Louis, Missouri, USA), or prepared by methods known to those skilled in the art according to the procedures described in the references, such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-15(John Wiley, Sons, 1991), Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and documents (Puvier science, 1989), Organic Chemistry, viscosity of carbon company, Inc. (John Wilcompany's, Inc, and company's, Inc. (Sound, Inc., and company's, Inc. (binder, Inc., and company's, Inc. (binder, Inc., and company's, Inc., binder, Inc., and company's, Inc., binder. These schemes are merely illustrative of some of the methods by which the compounds of the present invention can be synthesized, and various modifications can be made to these schemes, as will be suggested to one skilled in the art in view of the disclosure herein.

As will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing unwanted reactions. Protecting groups suitable for different functional groups and conditions suitable for protecting and deprotecting specific functional groups are well known in the art. For example, a number of protecting groups are described in t.w.greene and g.m.wuts, protecting groups in Organic Synthesis, Second Edition, Wiley, New York, 1991 and references cited therein.

If desired, the starting materials and intermediates of the reaction can be isolated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and the like. Such materials can be characterized using conventional means, including physical constants and spectral data.

The compounds of the invention will generally contain one or more chiral centers. Thus, such compounds may be prepared or isolated as pure stereoisomers, if desired. All such stereoisomers (and enriched mixtures) are included within the scope of the invention unless otherwise indicated. For example, pure stereoisomers (or enriched mixtures) may be prepared using optically active starting materials or stereoselective reagents well known in the art. Alternatively, racemic mixtures of such compounds can be separated, for example, using chiral column chromatography, chiral resolving agents, and the like.

Preparation of the Compounds of the invention

In general, to prepare the compounds of formula (I) of the present invention, an appropriately 7-substituted lincosamine (lincosamine) intermediate and an appropriately substituted pyrrolidinyl, piperidinyl, azetidinyl or azepane carboxylic acid are condensed under reaction conditions, preferably in an inert organic solvent, in the presence of a coupling agent and an organic base. This reaction can be carried out using any number of known coupling reagents, such as O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium Hexafluorophosphate (HATU), 1-hydroxybenzotriazole Hydrate (HOBT) with carbodiimide, isobutyl chloroformate, and the like. Suitable organic bases include Diisopropylethylamine (DIEA), Triethylamine (TEA), pyridine, N-methylmorpholine, and the like. Suitable inert organic solvents that may be used include, for example, N-dimethylformamide, acetonitrile, dichloromethane, and the like. This reaction is typically carried out using an excess of the carboxylic acid to the lincosamine, at a temperature in the range of from about 0 ℃ to about 50 ℃. The reaction continues until completion, which typically occurs in about 2 to 12 hours.

The appropriate 7-substituted lincosamine intermediate (i.e., R) as defined in the present invention²/R³) Synthesized from methyl 6-amino-6, 8-dideoxy-1-thio-erythro-alpha-D-galacto-pyranoside by methods well known to those skilled in the art, which can be prepared as described in Hoeksema, H.et al.journal of the American Chemical Society, 1967, 892448-2452. Illustrative syntheses of 7-substituted lincosamine intermediates are shown in schemes 1-6 below.

Also as defined in the invention of the appropriate 7-substituted lincosamine intermediates (i.e. R)²/R³) Synthesized from methyl 6-amino-6, 8-dideoxy-1-thio-erythro- α -D-galacto-octapyranoside by methods well known to those skilled in the art, as disclosed in the following references: U.S. Pat. No.3,086,912 (R)²＝OH，R³H), us patent No.3,496,136, us patent No.3,502,646 or preferably european patent No.0161794 (R)²Halogen, R³H), us patent No.3,179,565 (R)²＝SR，R³H), U.S. patent No.3,544,551(R²＝SH，R³＝H)。

An appropriately substituted pyrrolidinyl or piperidinyl carboxylic acid intermediate (i.e. R) as defined herein⁹) Also synthesized from proline and pyridine by methods well known to those skilled in the art. Proline and pyridine which may be used in the synthesis of the carboxylic acid intermediates of the present invention include, for example, 4-oxoproline and 4-substituted pyridine. The proline and pyridine used in the synthesis are available from commercial suppliers, such as Aldrich and Sigma. Alternatively, these prolines and pyridines may be prepared by methods well known in the art. Illustrative syntheses of appropriately substituted pyrrolidinyl or piperidinyl carboxylic acid intermediates are shown in schemes 7-12 below.

Scheme 1 below illustrates the general synthesis of lincosamine intermediate 1c, wherein P is an N-protecting group, preferably Cbz or Boc, R ¹Is as defined for formula (I).

Scheme 1: general Synthesis of lincosamine intermediate 1c

(a) N-protection (Boc, Cbz); (b) o-silyl protection (TMS); (c) swern oxidation

As shown in scheme 1, methyl 6-amino-6, 8-dideoxy-1-thio-erythro- α -D-galacto-octapyranoside 1a is prepared as described in Hoeksmem, H.et.al.journal of the American chemical Society, 1967, 89, 2448-2452. The amino and hydroxyl functions of product 1a are then protected with suitable protecting groups. Suitable N-protecting groups (P) can be formed by adding di-tert-butyl dicarbonate, N- (benzyloxycarbonyloxy) succinimide, and the like. The hydroxyl group may be protected as a silyl ether. The hydroxyl group can be converted to Trimethylsilyl (TMS) ether by reaction with N, O-bis (trimethylsilyl) trifluoroacetamide in the presence of a suitable organic base, such as triethylamine, or converted to trimethylsilyl chloride in the presence of an organic base, such as triethylamine. The N-protection is usually done before the O-protection. After evaporation of the solvent, the crude product is chromatographed on silica to give the protected product 1 b.

Chemoselective deprotection and oxidation of the 7-O-trimethylsilyl group of 1b affords 7-keto-lincosamine derivative 1 c. This selective conversion is carried out by adding the protected product 1b to dimethyl sulfoxide and oxalyl chloride in an inert organic solvent, such as dichloromethane, followed by a suitable organic base, such as triethylamine. Alternatively, the conversion may be carried out by adding 1b to dimethyl sulfoxide and a suitable activator, for example trifluoroacetic anhydride, in an inert organic solvent. The reaction is generally carried out at a temperature of about-70 ℃. The resulting reaction mixture was stirred at low temperature and then warmed to about-50 ℃. The reaction is maintained at this second temperature for about 1 to 3 hours. A suitable organic base, such as TEA, pyridine, and the like, is added to the reaction mixture. The reaction mixture is suitably worked up to give product 1 c. The general class of conditions used in the conversion of 1b to 1c is known in the art as Swern oxidation conditions.

Scheme 2 below illustrates the general synthesis of lincosamine intermediate 2b, where P is an N-protecting group, preferably Cbz or Boc, R³Is hydrogen, R²' and R²Consistently, as defined for formula (I), R¹Is as defined for formula (I).

And (2) a flow scheme: general Synthesis of lincosamine intermediate 2b

(a) Wittig olefination (R)²’CH₂Ph₃ ⁺X^-Or R²’CH₂PO(OEt)₂Base, solvent); (b) h₂(ii) Pd, (c) Global deprotection

As shown in scheme 2, byThe keto-lincosamine intermediate 1e is reacted with Wittig or Horner-Wadsworth-Emmons to form the alkene. In this reaction, the appropriate phosphonium salt or phosphonate is deprotected with a strong base to produce a phosphonium ylide. Suitable phosphonium salts which may be used are alkyltriphenylphosphonium halides which can be prepared by reaction of triphenylphosphine with an alkyl halide. Suitable phosphorus compounds include, for example, methyltriphenylphosphonium bromide, diethyl (cyanomethyl) phosphonate, and the like. Suitable strong bases that can be used to form the ylide include organolithium reagents, potassium tert-butoxide, and the like. The formation of phosphonium ylides is generally carried out under an inert atmosphere, for example N₂In inert organic solvents, such as toluene, THF, etc., at low temperatures.

After the formation of the phosphonium ylide, the product 1c is added to the reaction. The reaction may suitably be carried out at a temperature between-40 ℃ and room temperature, with stirring until completion, typically 1 to 4 hours. The resulting organic solution was treated and the crude product was chromatographed on silica to give the alkene product 2 a.

The product 2a is then hydrogenated to give a saturated product 2 b. Hydrogenation is typically carried out using 10% palladium on carbon in a Parr bottle in a polar organic solvent such as methanol, ethanol, and the like. Using the bottle H₂Purge and aerate to about 50 to 70psi, shake until complete, typically about 12 to 24 h. The resulting reaction mixture is filtered, for example through celite, and washed with a polar organic solvent, for example methanol. The organic solution was treated as follows and transferred to a column containing dried, washed Dowex 50w-400x H⁺Resin funnel of type, shake. After washing the resin with methanol and water, washing with MeOH containing 5% TEA, the product 2b was eluted from the resin. The product can also be purified by silica gel column chromatography.

Scheme 3 illustrates a general synthesis of lincosamine intermediate 3b, where P is an N-protecting group, preferably Cbz or Boc, R²Or R³One is alkyl and the other is-OH, R¹Is as defined for formula (I).

And (3) a flow path: general Synthesis of lincosamine intermediate 3b

(a)R²M (carbon nucleophile); (b) (i) TMS deprotection (H)⁺Or F^-) And (ii) N-deprotection

As shown in scheme 3, a suitable carbon nucleophile is added to 7-ketolincosamine intermediate 1c in a suitable inert organic solvent to provide 7-hydroxyllincosamine intermediate 3 b. Suitable carbon nucleophiles include methylmagnesium chloride, diethylzinc, sodium acetylene, and the like, and suitable inert organic solvents that can be used include THF, diethyl ether, toluene, and the like. The reaction is usually carried out under reduced pressure at about 0 ℃ for about 3 to 5 hours. The reaction is then quenched with a saturated aqueous acidic solution, such as saturated aqueous NH ₄Cl/H₂And O. The quenched mixture is then processed and may be purified by chromatography to provide product 3 b.

Scheme 4 below illustrates the general synthesis of lincosamine intermediate 4b, wherein P is an N-protecting group, preferably Boc, R¹Is as defined in formula (I), R²/R³Is oxime (═ NOR)⁷) Wherein R is⁷Is as defined for formula (I).

And (4) a flow chart: general Synthesis of 7-oxime-lincosamine 4b

(a)H₂NOR⁷HCl, pyridine, EtOH (b) TFA

As shown in scheme 4, lincosamine intermediate 1c is converted to an oxime by stirring in the presence of a suitable reagent, such as O-trimethylsilylhydroxylamine, O-alkylhydroxylamine hydrochloride (e.g., O-methylhydroxylamine hydrochloride), and the like. The reaction is usually carried out in a polar organic solvent, such as methanol. The reaction may be conveniently carried out at room temperature for about 8 to 24 h. The solvent is removed to afford the N-protected product 4 a.

Removal of the protecting group can be carried out with an acid, such as trifluoroacetic acid (TFA), hydrochloric acid, p-toluenesulfonic acid, and the like, in an inert organic solvent such as dichloromethane, dichloroethane, dioxane, THF, and the like. The removal is usually carried out at low temperature, for example 0 ℃ and then gradually warmed to room temperature to give the product 4 b.

Scheme 5 below illustrates the general synthesis of lincosamine intermediate 5b, wherein R ²And R³Are both fluorine, P is an N-protecting group, preferably Cbz or Boc, R¹Is as defined for formula (I).

And (5) a flow chart: general Synthesis of 7-deoxy-7, 7-difluorolincosamine 5b

(a)F^-；(b)Ac₂O, pyridine, DMAP; (c) DAST; (d) TFA

As shown in scheme 5, lincosamine intermediate 1c is contacted with a suitable fluoride in an inert organic solvent. Suitable fluorides that may be used include tetrabutylammonium fluoride, Amberlite resins A-26F^-Type, HF pyridine, etc. Suitable inert organic solvents include THF, acetonitrile, dichloromethane, dioxane, and the like. The reaction may suitably be carried out at room temperature for about 1 to 2 h. The product (not shown) can be purified over a silica gel column.

The O-protecting groups on the product obtained from the column are converted by contacting with acetic anhydride and Dimethylaminopyridine (DMAP) in a suitable mixture of an inert organic solvent and an organic base, such as dichloromethane and pyridine. The reaction may be suitably carried out at room temperature for about 6 to 12 hours. The product can be purified by silica gel column to give product 5 a.

The product 5a is contacted with a suitable fluorinating agent, and the N-protecting group is then removed to provide the product 5 b. Suitable fluorinating agents which may be used include, for example, dimethylaminosulfur trifluoride, [ bis (2-methoxyethyl) amino ] sulfur trifluoride, and the like. The reaction is usually carried out in an inert organic solvent, such as dichloromethane, ethyl acetate, THF, etc., at room temperature for about 6 to 12 h.

Removal of the protecting group can be carried out with an acid, such as trifluoroacetic acid (TFA), hydrochloric acid, p-toluenesulfonic acid, and the like, in an inert organic solvent such as dichloromethane, dichloroethane, dioxane, THF, and the like. Removal is typically carried out at low temperature, e.g., 0 ℃, and then gradually warmed to room temperature to provide product 5 b.

Scheme 6 below illustrates the general synthesis of lincosamine intermediate 6b, wherein P is an N-protecting group, preferably a trifluoroacetyl group, R²And R³One being hydrogen and the other being Cl, Br or I, R¹Is as defined for formula (I).

And (6) a flow path: general Synthesis of 7-deoxy-7-halolincosamines 6b

(a) Methyl trifluoroacetate, triethylamine; (b) halogenating agents (i.e. PPh)₃X₂Wherein X ═ Cl, Br, I or preferably 1-N- (halomethylene) piperidine salts); (c) aqueous alkali (i.e. KOH, ammonia)

As shown in scheme 6, lincosamine intermediate 1a is N-protected with a suitable trifluoroacetylating reagent in the presence of a base in a suitable organic solvent. Suitable trifluoroacetylating agents include methyl trifluoroacetate, ethyl trifluoroacetothioacetate, trifluoroacetic anhydride and the like. Suitable organic solvents include methanol, THF, acetonitrile, dichloromethane, dioxane, and the like. The reaction may suitably be carried out at ambient temperature for about 2 to 4 h. The protected lincosamide (lincosamide) intermediate 6a may be purified by crystallization or used directly in a subsequent reaction.

Halogenation of the 7-position of the protected intermediate 6a is accomplished by contacting with a suitable Rydon reagent, such as Magerlein, b.j.; kagen, F.journal of medicinal Chemistry, 1969, 12, 780-784, or with an amide halide salt, as described in European patent No. 0161794. Suitable Rydon reagents include triphenylphosphene dichloride, triphenylphosphene dibromide, and the like, in an inert organic solvent such as acetonitrile, dichloromethane, dichloroethane, or toluene. Suitable haloamide salt reagents include 1-N- (chloromethylene) piperidine chloride, 1-N- (chloromethylene) -N-methylmethanatinium chloride, and the like, in an inert organic solvent such as acetonitrile, dichloromethane, dichloroethane, or toluene. The reaction is generally carried out at a temperature of about 24 ℃ to 70 ℃ for a period of 16 to 24h, using an excess of halogenating agent. Hydrolysis of the halogenation product adduct (not shown) and removal of the protecting group in aqueous base affords 7-deoxy-7-halolincosamide intermediate 6 b. Suitable bases are solutions of NaOH, KOH and concentrated ammonia in water or mixtures of water and miscible organic solvents such as methanol, acetonitrile, tetrahydrofuran, dioxane, and the like. The reaction is typically carried out under conditions that precipitate the crude 7-deoxy-7-halolincosamide intermediate 6 b. The 7-deoxy-7-halolincosamide intermediate 6b may be purified by crystallization from a suitable solvent or solvent system.

Alternatively, 1c may be directly halogenated, as described in U.S. Pat. No.3,496,136 or U.S. Pat. No.3,502,646, in contact with a suitable Rydon reagent or amide halide salt, as described in European patent No. 0161794. Hydrolysis of the halide adduct (not shown) in aqueous base affords 7-deoxy-7-halolincosamide intermediate 6 b.

Scheme 7 below illustrates trans R⁹General Synthesis of "-proline intermediate 7d, where R is⁹"is alkyl or substituted alkyl.

Scheme 7: general Synthesis of Trans-alkylproline 7d

(a) (i) LiHMDS, THF-78 ℃, (ii) bromoene; (b) (i) LiBHEt₃，THF-78℃，(ii)BF₃·OEt₂，Et₃SiH；(c)H₂ Pd/C.

As shown in scheme 7, protected 5-oxoproline 7a is enolized with a suitable base in an inert organic solvent and then alkylated with a suitable alkylating agent to give lactam 7b (wherein R is⁹' is alkenyl), such as Zhang, r; the journal of the American chemical Society, 1998, 120, 3894-. Compound 7a is available from commercial suppliers, such as Bachem. Alternatively, 7a may be prepared by methods well known in the art. Suitable basic enolizing agents include LiHMDS, LiN (iPr)₂And the like, suitable alkylating agents include allylic and benzylic bromides such as 4-bromo-2-methyl-2-butene and cis-1-bromo-2-pentene, allyl bromide, and the like.

Reduction of lactam 7b with a suitable reducing agent gives pyrrolidine 7c, wherein R⁹' is an alkenyl group. The reduction being carried out by means of a two-step sequence involving lactamsReduction to give the hemiaminal, and subsequent reduction of the hemiaminal. Suitable reducing agents that may be used include Et₃SiH/BF₃·OEt₂、Et₃SiH/TiCl₄And the like.

Pyrrolidine 7c is then hydrogenated with removal of R⁹' unsaturation in the substituent and removal of the benzyl protecting group in the carboxylic acid gives the product 7 d. Hydrogenation is typically carried out in a polar organic solvent, such as methanol, ethanol, etc., using 10% palladium on carbon in a Parr bottle. Using the bottle H₂Purging and aerating to about 50 to 70psi, shaking until completionAnd usually about 5 to 24 hours. The reaction mixture is filtered, for example through a pad of celite, and washed with a polar organic solvent, for example methanol. The combined washings and filtrate were evaporated to give product 7d, wherein R⁹"is alkyl or substituted alkyl.

Scheme 8 below illustrates trans-R⁹General Synthesis of proline intermediates 8b and 8c, where R⁹' is alkenyl or substituted alkenyl, R⁹"is alkyl or substituted alkyl.

And (3) a process 8: trans-R⁹General Synthesis of substituted prolines 8b, where R⁹' is alkenyl, R ⁹Is "R⁹The saturated form of' and the synthesis of 8c, wherein R⁹' is alkenyl or substituted alkenyl

(a)i.O₃，DCM，-78℃，ii.DMS；(b)R⁹’CH₂P⁺Ph₃Salts, bases; (c) h₂，Pd/C；(d)Aq.LiOH，THF.

As shown in scheme 8, product 7c is ozonated to give an aldehyde, which is then treated under Wittig conditions to give 8 a. Ozonolysis reactions are generally carried out in anhydrous inert organic solvents, such as dichloromethane, dioxane, THF, etc., at low temperatures, e.g., -78 ℃, followed by quenching reactions with reducing agents, such as DMS, Ph₃P。

The aldehyde is reacted with a suitable phosphonium salt in an inert organic solvent in the presence of a strong base. Suitable phosphonium salts that may be used include, for example, fluorobenzylphosphonium chloride, 4-chlorobenzylphosphonium chloride, dibromofluoromethane, triphenylphosphine and the like. Suitable bases that can be used include potassium tert-butoxide, organolithium reagents and activated zinc. Suitable organic solvents that may be used include toluene, THF, dimethylacetamide, and the like. The reaction is generally carried out under an inert atmosphere, for example under nitrogen, with vigorous stirring. The reaction is typically carried out at room temperature to about 110 ℃ for 1 to 2 h. The resulting reaction mixture is suitably treated and may be purified by chromatography to afford 8 b.

Intermediate 8b is then hydrogenated to provide product 8 c. Hydrogenation is typically carried out in a polar organic solvent, such as methanol, ethanol, etc., using 10% palladium on carbon in a Parr bottle. Using the bottle H ₂Purge and aerate to about 40 to 70psi, shake until complete, typically about 4 to 24 h. The reaction mixture is filtered, for example through a pad of celite, and washed several times with a polar organic solvent, for example methanol. The combined washings and filtrate were evaporated to give product 8c, wherein R⁹"is alkyl or substituted alkyl, corresponding to the saturated form of product 8 b.

Alternatively, intermediate 8b can be saponified by methods well known to those skilled in the art, and contacted with an aqueous base and a miscible organic cosolvent to provide R⁹' unsaturated amino acid intermediate 8 c.

Scheme 9 below illustrates the general synthesis of proline intermediate 9d, wherein R⁹Is as defined for formula (I).

And (3) a process 9: cis-R⁹Or trans-R⁹General Synthesis of substituted Cyclic amino acids 9d

(a) Activating agents, i.e., (Ts)₂O, pyridine or PPh₃Br₂) (b) nucleophiles, bases (RSH, MTBE), DMF (c) LiOH, THF, H₂O

Scheme 10 below illustrates a general synthesis of substituted pyridinecarboxylic acid intermediates 10b, wherein R⁹Is as defined for formula (I), e.g. Shuman, r.t.; journal of organic chemistry.1990, 55, 741-750.

A process 10: general Synthesis of substituted pyridin-2-yl Carboxylic acids 10b

As shown in scheme 10, an appropriately substituted pyridine is contacted with a suitable oxidizing agent in an inert organic solvent. Appropriately substituted pyridine starting materials are available from commercial suppliers, such as Aldrich and Sigma. Alternatively, these pyridines may be prepared by methods well known in the art. Suitable oxidizing agents that may be used include hydrogen peroxide, MCPBA, and the like. The reaction is usually carried out under reflux for 6 to 12 h. The reaction mixture is then contacted with a suitable cyanide reagent to provide the cyano-substituted pyridine 10 a. Suitable cyanide reagents that may be used include trimethylsilyl cyanide, HCN, and the like. Suitable inert organic solvents include dichloromethane, dioxane, THF, and the like. The reaction may be conveniently carried out at room temperature for about 6 to 12 h. The reaction mixture was worked up to give cyano-substituted pyridine 10 a.

The cyano-substituted pyridine 10a is then hydrolyzed by contact with a suitable acid to provide pyridin-2-yl carboxylic acid 10 b. Suitable acids for hydrolyzing cyano groups to carboxylic acids include hydrochloric acid, aqueous sulfuric acid, and the like. The reaction is usually carried out under reflux for 6 to 12 h.

The following scheme 11 illustrates the general synthesis of pyridine and piperidine intermediates, wherein R⁹Is as defined for formula (I).

Scheme 11: general Synthesis of 4-substituted intermediates 11c, 11d and 11e

(a)SOCl₂MeOH (not shown); (b) HI, H₂PO₃，(c)MeOH，H₂SO₄(cat.)，(d)Pd(OAc)₂，CuI，PPh₃R⁹' alkyne; (e) PtO₂，H₂，H⁺(ii) a (f) N-protecting reagent (i.e., (Boc)₂O, cbzccl ect.), a base; (g) aq, lioh, dioxane

Scheme 12 below illustrates the general synthesis of proline intermediate 12d, wherein R⁹Is as defined for formula (I).

And (3) a process 12: general synthesis of proline intermediate 12d starting from 4-ketopyrrolidine (m ═ 1) and 4-ketopiperidine (m ═ 2)12a

(a) Tetraallyltin, BF₃·Et₂O or R⁹M(R⁹Carbon nucleophiles); (b) DAST; (c) h₂Deprotection conditions for Pd (d) aq. LiOH or suitable carboxylic acid esters

As shown in scheme 12, ketoproline 12a is allylated to produce hydroxy allylproline, the hydroxy function of which is subsequently replaced with fluorine. Hydrogenation of the allylic double bond affords fluoroalkyl proline 12c, which is deprotected to 12 d.

Scheme 13 below illustrates the coupling reaction of lincosamine intermediates prepared as described in schemes 1-6 above with pyrrolidinyl or piperidinyl carboxylic acids prepared as described in schemes 7-12 above, wherein R is¹、R²、R³、R⁶And R⁹Is as defined in formula (I), P¹Are suitable O-protecting groups, P²Are suitable N-protecting groups.

Scheme 13: general coupling and deprotection methods

As shown in scheme 13, an appropriately 7-substituted lincosamine intermediate (e.g., prepared according to any of schemes 1-6) is condensed with an appropriately substituted pyrrolidinyl or piperidinyl carboxylic acid (e.g., prepared according to any of schemes 7-9 or 11-12) under reaction conditions, preferably in an inert organic solvent, in the presence of a coupling reagent and an organic base. This reaction can be carried out using any number of known coupling reagents, such as O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium Hexafluorophosphate (HATU), 1-hydroxybenzotriazole Hydrate (HOBT) with carbodiimide, isobutyl chloroformate, and the like. Suitable organic bases include Diisopropylethylamine (DIEA), Triethylamine (TEA), pyridine, N-methylmorpholine, and the like. Suitable inert organic solvents that may be used include, for example, N-dimethylformamide, acetonitrile, dichloromethane, and the like. This reaction is typically carried out using an excess of the carboxylic acid to the lincosamine at a temperature of from about 0 ℃ to about 50 ℃. The reaction continues until completion, which typically occurs in about 2 to 12 hours.

Removal of the protecting group can be carried out with an acid, such as trifluoroacetic acid (TFA), hydrochloric acid, p-toluenesulfonic acid, and the like, in an inert organic solvent such as dichloromethane, dichloroethane, dioxane, THF, and the like. The removal is usually carried out at a low temperature, for example 0 ℃, and then gradually warmed to room temperature to give the product.

Also shown in scheme 13, an appropriately 7-substituted lincosamine intermediate (e.g., prepared according to any of schemes 1-6) is condensed with an appropriately substituted pyridin-2-yl carboxylic acid (e.g., prepared according to scheme 10) under reaction conditions, preferably in an inert organic solvent, in the presence of a coupling reagent and an organic base, as described above.

Pyridine 13b was hydrogenated to give the piperidinyl product. Hydrogenation is typically carried out using platinum (IV) oxide in a polar organic solvent, such as methanol, ethanol, and the like, in the presence of an acid, such as HCl, acetic acid, and the like, in a Parr bottle. Using the bottle H₂Purge and aerate to about 40 to 70psi, shake until complete, typically about 24 h. The reaction mixture is filtered, for example through a pad of celite, washed several times with a polar organic solvent,such as methanol. The washings and filtrate were combined and evaporated to give the piperidinyl product.

Coupling of pyridine carboxylic acids with lincosamines to give pyridine 13b, followed by reduction to piperidinyl products may also be performed as in Birkenmeyer, R.D; et al; journal of medicinal chemistry 1984, 27, 216-.

Scheme 14 below illustrates the coupling reaction of lincosamine intermediates prepared as described in schemes 1-6 above with pyrrolidinyl or piperidinyl carboxylic acids prepared as described in schemes 7-12 above, wherein R is¹、R²、R³And R⁹Is as defined for formula (I) and P is a suitable N-protecting group. The coupling reactions described herein may also be used to couple azetidinyl and azepane carboxylic acids.

The process 14: general Synthesis of 4-thioether lincosamide 14c

(a)(TEA，CF₃COOEt)(b)MTL，BSTFA，TEA，HATU(c)Dowex H⁺Resin MeOH (d) (Ac)₂O, pyridine, DMAP (e) TFA, DMS, DCE, H₂O(f)(Ts)₂O, pyridine, DCM (g) R⁹H, MTBE (wherein R⁹Is selected so as to introduce alkylthio or substituted alkylthio substituents; sulfoxide or sulfone substituents are also within the scope of the invention and can be obtained by conventional oxidation methods well known in the art) (h) MeONa, MeOH

Scheme 15 illustrates a general synthetic method for the construction of protected 1-allylic intermediates 15b, 15c, 15e, 15f, wherein R²、R³、R⁹Is as defined in formula (I), P₁And P₂Respectively, represent suitable N-and O-protecting groups.

Scheme 15: general Synthesis for the construction of protected 1-allylic intermediates 15b, 15c, 15e, 15f

(a)DAST，NBS(b)BF₃·Et₂O，CH₃CN，TMS，CH₂CH＝CH₂

In scheme 15, P₁And P₂Are protecting groups which can preferably be removed. Replacement of the methylthio (methylsulfanyl) group with a fluoro substituent is accomplished by contacting with DAST in the presence of N-bromosuccinimides (NBs) in a suitable solvent, such as Dichloromethane (DCM), to provide compounds 15b and 15 c.

Further, the fluoro group is displaced by contacting with trimethyallylsilane in the presence of a trifluoroborate diethyl ether complex to generate an allyl substituent. Subsequent removal of the Boc (t-butyloxycarbonyl) protecting group with trifluoroacetic acid (TFA) provided the deprotected product. Alternatively, conventional defluorination of 1-des (methylthio) -1-fluoro-2, 3, 4-tri-O-benzyl-7-deoxy-7-methylinkinamine gives 1-des (methylthio) -2, 3, 4-tri-O-benzyl-7-deoxy-7-methylinkinamine (that is, R¹Is hydrogen).

The conventional amide coupling of the carboxyl group of N-Boc-4-pentyl-proline with 1-des (methylthio) -1-allyl-2, 3, 4-tri-O-benzyl-7-deoxy-7-methylinkinamide in Dimethylformamide (DMF) and Triethylamine (TEA) in the presence of a coupling promoter, such as O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium Hexafluorophosphate (HATU), gives compounds 15c and 15 f. The allyl groups of compounds 15c and 15f provide a source of extensive modification of the 1-position of the lincosamine groups.

Scheme 16 illustrates a general synthetic approach to construction wherein R¹Is alkylthio, substituted alkylthio, R²、R³、R⁹Is as defined in formula (I), P₁And P₂Respectively, represent suitable N-and O-protecting groups.

And (3) a process 16: replacement of fluoro groups by sulfanyl moieties

Scheme 16 illustrates that nucleophilic displacement of the 1-fluoro group by a suitable sulfanyl moiety can be accomplished either on the lincosamine moiety to yield compound 16a or on the coupled lincosamine derivative to yield compound 16 b. Nucleophilic displacement is carried out using conventional techniques well known in the art.

Scheme 17 illustrates a general synthetic method for constructing alcohol and ether substituents at the 1-position, wherein R²、R³、R⁹Is as defined in formula (I), P₁And P₂Each represents a suitable N-and O-protecting group, and R is hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl.

Scheme 17: general Synthesis of alcohol and Ether 1-position Lincolamine derivatives

(a)i.O₃-78 ℃ to 0 ℃, ii. NaBH₄(b) RX, a base, wherein R can be selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl

Scheme 17 illustrates that 1- (desulfonylmethyl) -1-allyllincosamine derivatives can be oxidized to the corresponding aldehydes by conventional methods, which are then reduced to primary alcohols, e.g., ozonolysis followed by reduction with sodium borohydride, preferably in a protic solvent, e.g., methanol. Subsequently, the primary alcohol is contacted with a suitable base, such as sodium hydride and a suitable alkyl halide, to produce an ether derivative, either a lincosamine entity or a coupled lincosamine derivative, to provide compounds 17c and 17f, respectively.

Scheme 18 illustrates the deprotection schemes of 15f, 16b and 17f, where R²、R³、R⁹Is as defined for formula (I), P is a suitable N-protecting group, R¹Which are consistent with schemes 15, 16 and 17, respectively.

Scheme 18

Scheme 18 illustrates conventional deprotection to provide compounds of formula (I).

Scheme 19 below illustrates the alkylation of a pyrrolidinyl or piperidinyl ring nitrogen wherein R⁶Is alkyl or hydroxyalkyl, R¹、R²、R³And R⁹Is as defined for formula (I).

Scheme 19: general Synthesis of 1' -N-substituted lincosamides

(a) Alkylating agents

As shown in scheme 19, lincosamine 18a can be N-substituted by contacting with an alkylating agent in the presence of a suitable base to provide product 18 b. Suitable alkylating agents that may be used include epoxides, alkyl bromides, and the like. Suitable bases that may be used include potassium carbonate, cesium carbonate, triethylamine and the like. The alkylation reaction is usually carried out in a polar organic solvent, such as methanol or DMF. The alkylation reaction is generally carried out at a low temperature of from 0 ℃ to-10 ℃ for from 10 to 20 hours.

In scheme 20, R²、R³、R⁶And R⁹Is as defined in formula (I), P₂Are suitable O-protecting groups.

Scheme 2O: rainey Ni, EtOH, reflux

Scheme 21 below illustrates a versatile synthetic sequence that allows for the synthesis of an unsaturated N-protected amino acid 21k ring, wherein m and R ⁹Is as defined for formula (I).

(a)NaH，R⁹Br，DMF(b)KOH，H₂O，EtOH(c)CH₂O (aq), piperidine, EtOH (d) DIBALH, CH₂Cl₂(e)PBr₃，Et₂O (f) ethyl bromoacetate (g) HCl/dioxane (h) LiOt-Bu, THF (i)21d, LiHMDS, LiCl, THF, 0 ℃ (j) Boc₂O，Et₃N，CH₂Cl₂(k) Olefin metathesis catalyst CH₂Cl₂(l)1M NaOH(aq)，MeOH

As shown in scheme 21, a suitable N-allylic amino ester 21f can additionally serve as a pseudoephedrine chiral auxiliary, allowing stereospecific alkylation of the carbon by a suitable allylic bromide 21 d. Protection of the secondary amine followed by olefin metathesis and cleavage of the chiral auxiliary affords the 4, 5-unsaturated N-protected cyclic amino acid 21 k.

The process 22: a.H⁺MeOH b.2-Nitrobenzenesulfonyl chloride, 2, 4, 6-trimethylpyridine, dichloroethane c.Cs₂CO₃TBABr, DMF, 22c (Y ═ Br or OTs) or c₃Diisopropylazidodicarboxylate, 22c (Y ═ OH) d. benzylidene [1, 3-bis (2, 4, 6-trimethylphenyl) -2-imidazolidinylidene]Dichloro- (tricyclohexylphosphine) ruthenium e. organic base (7-methyl-1, 5, 7-triazabicyclo [4.4.0]Dec-5-ene), thiophenol f. (Boc)₂O, TEA g.aq.lioh, dioxane

Flow 23: general Synthesis of C6 lincosamine derivatives

(a)R²⁰+R²¹Metal, Et₂AlCl，Et₂O；(b)(i)MsCl，Et₃N，CH₂Cl₂(ii) Giralard's reagent T, MeOH, (iii) trifluoroacetic anhydride, 2, 6-lutidine, CH ₂Cl₂；(c)(i)TFA，H₂O，(ii)Ac₂O，Et₃N，DMAP，CH₂Cl₂，(iii)HBr，AcOH，CH₂Cl₂；(d)(i)AcOAg，AcOH，(ii)PCl₅，BF₃.OEt₂，CH₂Cl₂；(e)(i)MeSNa，HMPA，DMF，(ii)Ac₂O，Et₃N，DMAP，CH₂Cl₂；(f)NaOH，H₂O，MeOH

The process 24: general Synthesis of 2-substituted esters

a.(Boc)₂O，aq.KHCO₃THF b p-anisaldehyde dimethyl acetal, PPTS, c.R¹¹Acylating agent, base d.TFA, DCE, water

Flow 25: general Synthesis of trans-alkyl azetidinecarboxylic acids

(a) (i) LDA, THF, 0 ℃, (ii) alkyl or alkenyl bromides; (b) TMSCHN₂，MeOH，23℃；(c)(i)TMSCHN₂，MeOH，23℃，(ii)H₂，Pd/C，EtOAc，23℃；(d)Et₃N.3HF，THF，23℃；(e)LiAlH₄，THF，68℃；(f)Boc₂O，CH₂Cl₂，23℃；(g)RuCl₃.xH₂O，NaIO₄Acetone, H₂O，23℃

The process 26: general synthesis of trans-alkyl azetidinecarboxylic acids via aldehydes

(a) TBSCl, imidazole, DMF, 23 ℃; (b) (i) ozone, CH₂Cl₂，-78℃，(ii)PPh₃(ii) a (c) Olefination of R₁R₂CHP^＝Ph₃X^-Base, solvent; (d) h₂Pd/C, EtOAc, 23 ℃ or KO₂CN＝NCO₂K, AcOH, dioxane, 23 ℃; (e) TBAF, THF, 23 ℃; (f) RuCl₃.xH₂O，NaIO₄Acetone, H₂O，23℃

The process 27: general Synthesis and resolution of racemic cis 4-substituted intermediate 27a to give 2S, 4R R⁹Intermediate 27b

(a)PtO₂，H₂，H⁺(b)(Boc)₂O，^-OH (c) i. chiral amine, recrystallized ii.H⁺

The process 28: general Synthesis of 2-substituted esters

a.(Boc)₂O，aq.KHCO₃THF b p-anisaldehyde dimethyl acetal, PPTS, c.R¹¹Acylating agent, base d.TFA, DCE, water (method V)

Pharmaceutical preparation

When used as a medicament, the compounds of the invention are generally administered in the form of a pharmaceutical composition. These compounds can be administered by a variety of routes including oral, parenteral, transdermal, intravenous, intramuscular, topical, rectal and intranasal. Both injectable and oral compositions of these compounds are effective. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.

The invention also includes pharmaceutical compositions comprising one or more of the above-described compounds of the invention as active ingredients, together with one or more pharmaceutically acceptable carriers. In preparing the compositions of the present invention, the active ingredient is typically mixed with an excipient, diluted with an excipient, or enclosed within such a carrier, which may be in the form of a capsule, sachet, paper or other container. The excipients employed are generally excipients suitable for administration to humans or other mammals. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material that serves as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments (containing, for example, up to 10% by weight of the active compound), soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

In preparing the formulation, it may be necessary to grind the active compound prior to combining with the other ingredients to provide the appropriate particle size. If the active compound is substantially insoluble, it is generally milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size is normally adjusted by milling to provide a substantially uniform distribution in the formulation, for example about 40 mesh.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. The formulation may additionally include: lubricants, such as talc, magnesium stearate and mineral oil; a humectant; emulsifying and suspending agents; preservatives, such as methyl and propyl hydroxybenzoate; a sweetener; and a flavoring agent. The compositions of the present invention are formulated to provide rapid, sustained or delayed release of the active ingredient after administration to the patient using techniques known in the art.

The amount of active ingredient, i.e., the compound according to the invention, in the pharmaceutical composition and its unit dosage form may be varied or adjusted depending upon the particular application, the potency of the particular compound and the desired concentration.

The compositions are preferably formulated in unit dosage forms containing from about 5 to about 100mg, more usually from about 10 to about 30mg, of the active ingredient per dose. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for humans and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Preferably, no more than about 20% by weight of the pharmaceutical composition of the above-described compounds of the present invention are employed, more preferably no more than about 15% by weight, with the balance being pharmaceutically inert carriers.

The active compounds are effective over a wide dosage range, generally being administered in a pharmaceutically or therapeutically effective amount. It will be understood, however, that the actual amount of the compound administered will be determined by a physician in the light of the relevant circumstances, including the condition being treated, the severity of the bacterial infection being treated, the chosen route of administration, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, and the like.

In therapeutic use in the treatment or combating of bacterial infections in warm-blooded animals, the compounds or pharmaceutical compositions thereof will be administered orally, topically, transdermally and/or parenterally in doses to achieve and maintain a concentration, i.e. the amount or blood level, at which the active ingredient will be antibacterially effective in the animal being treated. Generally, such an antibacterial or therapeutically effective amount (i.e., an effective dose) of the active ingredient will be in the range of about 0.1 to about 100, more preferably about 1.0 to about 50mg/kg body weight/day.

For preparing solid compositions, such as tablets, the principal active ingredient is mixed with a pharmaceutical excipient to form a pre-solid formulation composition containing a homogeneous mixture of the compound of the present invention. When referring to these preformulation compositions as homogeneous, this means that the active ingredient is dispersed evenly throughout the composition so that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This solid preformulation composition is then subdivided into unit dosage forms of the type described above, e.g. containing from 0.1 to about 500mg of the active ingredient of the invention.

The tablets or pills of the invention may be coated or otherwise compounded to provide a long-acting dosage form. For example, a tablet or pill may comprise an inner dosage and an outer dosage component, the latter being in the form of a capsule of the former. The two components may be separated by an enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials may be used for such enteric layers or coatings, such materials including various polymeric acids and mixtures of polymeric acids with shellac, cetyl alcohol and cellulose acetate, among others.

Liquid forms in which the novel compositions of the present invention may be incorporated for oral or injectable administration include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils such as corn oil, cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

Inhalation or insufflation compositions include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients, as described above. Preferably, the compositions are administered via the oral or nasal respiratory route for local or systemic effect. Compositions in preferred pharmaceutically acceptable solvents may be nebulized with an inert gas. The nebulized solution may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a face mask or intermittent positive pressure ventilator. The solution, suspension or powder composition may preferably be administered orally or nasally from a device that delivers the formulation in a suitable manner.

The following formulation examples illustrate representative pharmaceutical compositions of the present invention.

Formulation example 1

Hard gelatin capsules were prepared containing the following ingredients:

component amount (mg/capsule)

Active ingredient 30.0

Starch 305.0

Magnesium stearate 5.0

The above ingredients were mixed and filled into hard gelatin capsules at 340 mg.

Formulation example 2

Tablets were prepared using the following ingredients:

component amount (mg/tablet)

Active ingredient 25.0

Microcrystalline cellulose 200.0

Colloidal silica 10.0

Stearic acid 5.0

The ingredients were blended and compressed into tablets, each weighing 240 mg.

Formulation example 3

Preparing a dry powder inhalation formulation comprising:

the weight of the ingredients

Active ingredient 5

Lactose 95

The active ingredient is mixed with lactose and the mixture is added to a dry powder inhaler device.

Formulation example 4

Tablets, each containing 30mg of active ingredient, were prepared as follows:

component amount (mg/tablet)

Active ingredient 30.0mg

Starch 45.0mg

Microcrystalline cellulose 35.0mg

Polyvinylpyrrolidone (10% sterile aqueous solution) 4.0mg

Sodium carboxymethyl starch 4.5mg

Magnesium stearate 0.5mg

Talc 1.0mg

A total of 120mg

The active ingredient, starch and cellulose were passed through a No.20 mesh u.s. sieve and mixed thoroughly. The polyvinylpyrrolidone solution was mixed with the resulting powder and then passed through a 16 mesh u.s. sieve. The resulting granules were dried at 50 ℃ to 60 ℃ and passed through a 16 mesh u.s. sieve. Sodium carboxymethyl starch, magnesium stearate and talc, which had previously passed through a No.30 mesh U.S. sieve, were then added to the granules, which were mixed and compressed on a tablet press to obtain tablets each weighing 120 mg.

Formulation example 5

Capsules, each containing 40mg of drug, were prepared as follows:

component amount (mg/capsule)

Active ingredient 40.0mg

Starch 109.0mg

Magnesium stearate 1.0mg

A total of 150.0mg

The active ingredient, starch and magnesium stearate were blended and passed through a No.20 mesh u.s. sieve and filled into hard gelatin capsules at 150 mg.

Formulation example 6

Suppositories, each containing 25mg of active ingredient, are prepared as follows:

amount of ingredients

Active ingredient 25mg

Saturated fatty acid glycerides to 2,000mg

The active ingredient is passed through a No.60 mesh U.S. sieve, suspended in previously melted saturated fatty acid glycerides, and slightly heated if necessary. The mixture was then poured into suppository molds of nominal 2.0g capacity and cooled.

Formulation example 7

Suspensions were prepared as follows, containing 50mg of drug per 5.0mL dose:

amount of ingredients

Active ingredient 50.0mg

Xanthan gum 4.0mg

Sodium carboxymethylcellulose (11%)

Microcrystalline cellulose (89%) 50.0mg

Sucrose 1.75g

Sodium benzoate 10.0mg

Flavors and colors q.v.

Purified water to 5.0mL

The active ingredient, sucrose and xanthan gum were blended, passed through a No.10 mesh u.s. sieve and then mixed with a previously prepared aqueous solution of microcrystalline cellulose and sodium carboxymethylcellulose. The sodium benzoate, flavor and color are diluted with some water and added while stirring. Sufficient water was then added to bring the volume to the desired volume.

Formulation example 8

Component amount (mg/capsule)

Active ingredient 15.0mg

Starch 407.0mg

Magnesium stearate 3.0mg

A total of 425.0mg

The active ingredient, starch and magnesium stearate were blended and passed through a No.20 mesh u.s. sieve and filled into hard gelatin capsules at 425.0 mg.

Formulation example 9

Subcutaneous formulations can be prepared as follows:

amount of ingredients

Active ingredient 5.0mg

Corn oil 1.0mL

Formulation example 10

Topical formulations can be prepared as follows:

amount of ingredients

1-10g of active ingredient

Emulsifying wax 30g

20g of liquid paraffin

White soft paraffin to 100g

The white soft paraffin was heated until melted. Liquid paraffin and emulsifying wax are added and stirred until dissolved. The active ingredient is added and stirring is continued until dispersed. The mixture was then cooled until solid.

Formulation example 11

Intravenous formulations can be prepared as follows:

amount of ingredients

250mg of active ingredient

Isotonic saline 1000mL

Another preferred formulation for use in the method of the present invention employs a transdermal delivery device ("patch"). Such transdermal patches may be used to provide continuous or discontinuous infusion of a compound of the invention in controlled amounts. The construction and use of transdermal patches for drug delivery are well known in the art. See, for example, U.S. Pat. No.5,023,252 to 1991, issued on Ser. No. 6 and 11, incorporated herein by reference. Such patches may be configured for continuous, pulsatile, or on-demand delivery of the drug.

It is often necessary or desirable to introduce pharmaceutical compositions directly or indirectly into the brain. Direct techniques typically involve placing a drug delivery catheter in the host ventricular system to bypass the blood-brain barrier. One such implantable delivery system for the transport of biological agents to specific anatomical regions of the body is described in U.S. patent No. 5,011,472, which is incorporated herein by reference.

Indirect techniques are generally preferred and generally involve formulating a composition to provide latency of the drug to the fat-soluble drug by converting the hydrophilic drug. Latency is generally achieved by blocking hydroxyl, carbonyl, sulfate, and primary amine groups present on the drug to render the drug more lipid soluble and amenable to transport across the blood-brain barrier. Alternatively, intra-arterial infusion with hypertonic solutions may enhance delivery of hydrophilic drugs, which may temporarily open the blood-brain barrier.

Other suitable formulations for use in the present invention may be found in Remington's pharmaceutical sciences, machine Publishing Company, Philadelphia, PA, 17th ed. (1985).

As noted above, the compounds described herein are suitable for use in the various drug delivery systems described above. In addition, to increase the in vivo serum half-life of the administered compound, the compound may be encapsulated, introduced into a liposomal cavity, made into a colloid, or other conventional techniques may be employed to increase the serum half-life of the compound. Various methods can be used to prepare liposomes, such as described in Szoka, et al, U.S. Pat. Nos. 4,235,871, 4,501,728, and 4,837,028, each of which is incorporated herein by reference.

As noted above, the compound administered to the patient is in the form of a pharmaceutical composition as described above. These compositions may be sterilized by conventional sterilization techniques, or may be sterile filtered. The resulting aqueous solution may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation will generally be between 3 and 11, more preferably 5 to 9, most preferably 7-8. It will be appreciated that the use of certain of the aforementioned excipients, carriers or stabilizers will result in the formation of a pharmaceutical salt.

In general, the compounds of the present invention will be administered in a therapeutically effective amount by any of the accepted modes of administration for drugs that serve similar utilities. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining LD₅₀(dose lethal to 50% of the population) and ED₅₀(dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as LD₅₀/ED₅₀The ratio of. Compounds that exhibit a large therapeutic index are preferred.

Data obtained from cell culture assays and animal studies can be utilized in formulating a dosage range for use in humans. The dosage of such compounds is preferably such that the ED is included ₅₀With little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. As to any compound used in the method of the invention, it may be first cultured from cellsTherapeutically effective doses are estimated in assays. A dose can be formulated in animal models to achieve inclusion of IC based on cell culture assays₅₀(concentration of test compound that achieves half-maximal inhibition of symptoms). Such information can be used to more accurately determine the dosage available to a human. Plasma levels can be measured, for example, by means of high performance liquid chromatography.

Practicality of use

The compounds, prodrugs and pharmaceutically acceptable salts thereof, as defined herein have activity against at least one of a variety of bacteria, protozoa, fungi and parasites. For example, the compounds, prodrugs and pharmaceutically acceptable salts thereof may be active against gram positive and gram negative bacteria. The compounds, prodrugs, and pharmaceutically acceptable salts thereof may be active against a variety of fungi, including mucor and candida fungi, such as trichoderma racemosum or candida albicans. The compounds, prodrugs and pharmaceutically acceptable salts thereof may be active against a variety of parasites, including malaria and cryptosporidium parasites.

The compounds of the invention may exhibit activity against at least one of a variety of bacterial infections, including, for example, gram-positive infections, gram-negative infections, mycobacterial infections, mycoplasma infections, and chlamydia infections.

Because the compounds of the present invention may exhibit potent activity against a variety of bacteria, such as gram-positive bacteria, the compounds of the present invention may be useful antimicrobial agents that are effective against at least one of a variety of human and veterinary pathogens, including gram-positive bacteria. Gram-positive organisms for which the compounds of the invention may be effective include, for example, Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, enterococcus faecalis, enterococcus faecium, Haemophilus influenzae, Moraxella catarrhalis, Escherichia coli, Bacteroides fragilis, Bacteroides thetaiotaomicron, Clostridium difficile, and the like.

The compounds of the present invention may be combined with one or more additional antibacterial agents. One or more additional antibacterial agents may be active against gram-negative bacteria. One or more additional antibacterial agents may be active against gram positive bacteria. The combination of a compound of the present invention with one or more additional antibacterial agents may be used to treat gram-negative infections. The combination of a compound of the present invention with one or more additional antibacterial agents may be used to treat gram-positive infections. Combinations of the compounds of the present invention with one or more additional antibacterial agents may also be used to treat mycobacterial infections, mycoplasma infections or chlamydial infections.

The in vitro activity of the compounds of the invention can be assessed by means of standard test procedures, for example by determining the Minimum Inhibitory Concentration (MIC) by means of the agar Dilution method, as described in "applied Standard. methods for Dilution of analytical Suscientific results for bacteria that is the same as the group of bacteria that is found in the group of microorganisms," 3rd ed., published 1993 by the national Committee for Clinical Laboratory standards, Villanova, Pennsylvania, USA.

The amount administered to a mammalian patient will vary depending upon the drug being administered, the purpose of the administration (e.g., prevention or treatment), the condition of the patient, the mode of administration, and the like. For example, the composition is administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. An amount sufficient to accomplish this is defined as a "therapeutically effective amount". An effective amount for such use will depend on the disease condition being treated and the judgment of the attending physician in light of the severity of the inflammation, the age, weight and general condition of the patient, and like factors.

The composition to be administered to the patient is in the form of a pharmaceutical composition as described above. These compositions may be sterilized by conventional sterilization techniques, or may be sterile filtered. The resulting aqueous solution may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation will generally be between about 3 and about 11, more preferably from about 5 to about 9, most preferably from about 7 to about 8. It will be appreciated that the use of certain of the aforementioned excipients, carriers or stabilizers will result in the formation of a pharmaceutical salt.

The therapeutic dosage of the compounds of the invention will vary with, for example, the particular use being treated, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. For example, for intravenous administration, the dose will generally be in the range of from about 20 μ g to about 500 μ g per kilogram of body weight, preferably from about 100 μ g to about 300 μ g per kilogram of body weight. Suitable dosages for intranasal administration will generally range from about 0.1mg to 1mg per kilogram of body weight. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

The following synthetic and biological examples are provided to illustrate the invention and are not to be construed as limiting the scope of the invention in any way.

Examples

In the discussion above and in the examples below, the following abbreviations have the following meanings. If an abbreviation is not defined, it has a generally accepted meaning.

7-methyl-MTL ═ 1-methylthio-7-deoxy-7-methylinkinamine

Ac ═ acetyl group

apt is a clear triplet

Aq ═ water

atm ═ atmospheric pressure

Bn ═ benzyl

Boc ═ tert-butoxycarbonyl protecting group

broad singlet

BSTFA ═ N, O-bis (trimethylsilyl) trifluoroacetamide

¹³C NMR ＝ ¹³Carbon nuclear magnetic resonance

Cbz ═ carbonyloxybenzyloxy protecting group

CDCl₃Deuterated chloroform

CD₃OD-deuterated methanol

CD₃SOCD₃Deuterium-substituted dimethyl sulfoxide

cfu is a colony forming unit

D₂Deuterium substituted O

d ═ double peak

DAST ═ dimethylaminosulfur trifluoride

double peak

double bimodal with double bimodal dddd

DIBALH ═ diisobutylaluminum hydride

dt-doublet trimodal

DCE ═ dichloroethane

DCM ═ dichloromethane

DIEA is diisopropylethylamine

DMAP ═ dimethylaminopyridine

DMF ═ dimethylformamide

DMS (dimethyl sulfide)

DMSO ═ dimethyl sulfoxide

DPPA ═ diphenylphosphoryl azide

ED₅₀As dose therapeutically effective in 50% of the population

equiv ═ equivalent

ESMS (electrospray ionization mass spectrometry)

Et is ethyl

EtOAc ═ ethyl acetate

Et₂Diethyl ether (O ═ C)

g is g ═ g

h is hour

HATU ═ O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate

Acid salts

HOBT ═ 1-hydroxybenzotriazole hydrate

1H NMR spectrum

HPLC ═ high pressure liquid chromatography

Hz-Hz

IC₅₀Concentration of test compound to achieve half-maximal inhibition of symptoms

J ═ coupling constant in hertz

L is liter

LD₅₀Dose lethal to 50% of the population

Lithium hexamethyldisilazide (LiHMDS)

m is multiplet

M is molar concentration

MCPBA ═ 3-chloroperbenzoic acid

Me is methyl

MeCN ═ acetonitrile

MeOH ═ methanol

mg ═ mg

MHB (Mueller Hinton broth)

MHz-megahertz

MIC (minimum inhibitory concentration)

min is minutes

mL to mL

mm-mm

mmHg-mmHg

mmol ═ mmol

Ms (espos) ═ mass spectrometry (positive mode electrospray ionization)

Ms (esneg) mass spectrum (negative mode electrospray ionization)

MTBU ═ 7-methyl-1, 5, 7-triazabicyclo [4.4.0] dec-5-ene

MTL ═ 1-methylthiolincosamine (methyl 6-amino-6, 8-dideoxy-1-thio-erythro)

-alpha-D-galacto-pyranoside)

N is positive

NBS ═ N-bromosuccinimide

NMR (nuclear magnetic resonance)

OBz ═ benzyloxy protecting group

OtBu ═ t-butoxy

Pd/C ═ palladium/carbon

pg ═ picogram

Ph ═ phenyl

Pro ═ L-proline

psi pounds per square inch

q is quartet

q.v. ═ quantitative

R_fRetention factor

rt-room temperature

s ═ singlet

sat. (saturation)

t is triplet

TCI ＝ TCI America

TEA ═ triethylamine

TFA ═ trifluoroacetic acid

THF ═ tetrahydrofuran

TLC ═ thin layer chromatography

TMS ═ trimethylsilyl

Ts ═ tosyl group

Microgram of mug

μ L ═ microliter

Micromolar concentration of

v/v-volume/volume

In addition, the term "Aldrich" means that the compounds or reagents used in the latter process are commercially available from Aldrich Chemical Company, Inc., 1001 West Saint Paul Avenue, Milwaukee, WI 53233 USA; the term "Fluka" means that the compound or reagent is commercially available from Fluka Chemical Corp., 980 South 2nd Street, Ronkonkoma NY 11779 USA; the term "Lancaster" means that the compound or agent is commercially available from Lancaster Synthesis, inc., p.o. box 100 Windham, NH 03087 USA; the term "Sigma" means that the compound or reagent is commercially available from Sigma, p.o. box 14508, st.louis MO 63178 USA; the term "Chemservice" means that the compound or reagent is commercially available from Chemservice inc., Westchester, PA, USA; the term "Bachem" means that the compound or reagent is commercially available from Bachem Bioscience Inc., 3700 Horizon Drive, Renaissance at Gulph Mills, King of Prussia, PA 19406 USA; the term "Maybridge" means that the compound or agent is commercially available from Maybridge chemical co. trevillett, Tintagel, Cornwall PL34 OHW united kingdom; the term "RSP" means that the compound or agent is commercially available from RSP AminoAcid Analogs, inc., 106 South St., Hopkinton, MA 01748, USA; the term "TCI" means that the compound OR reagent is commercially available from TCI America, 9211North harborate St., Portland, Oregon, 97203, OR, USA; the term "Toronto" means that the compounds or reagents are commercially available from Toronto Reasearch Chemicals, Inc., 2 Brisbane Rd., New York, ON, Canada M3J2J 8; the term "Alfa" means that the compound or agent is commercially available from Johnson Matthey catalog Company, Inc.30 Bond Street, Ward Hill, MA 01835-; the term "Nova Biochem" means that the compounds or reagents are commercially available from Nova Biochem USA, 10933 North Torrey pins Road, P.O.Box 12087, La Jolla CA 92039-.

In the following examples, all temperatures are in degrees Celsius (unless otherwise indicated), and the compounds shown are prepared using the following general procedure.

General process

Method A

Methyl 6-amino-6, 8-dideoxy-1-thio-erythro- α -D-galacto-octapyranoside 1a (MTL) is prepared as described in Hoeksmem, H.et al.journal of the American chemical society, 1967, 89, 2448-. N- (benzyloxycarbonyloxy) succinimide (5.8g, 23.1mmol) and 1a (5.0g, 19.7mmol) were suspended in pyridine (40mL) in N₂Stirring for 36h under atmosphere. The reaction mixture was cooled to 0 ℃ and then bis-N, O-trifluoroacetamide (15.7mL, 59.0mmol) was added via syringe over 2 min. The reaction mixture was warmed to rt and stirred for 42 h. Toluene (100mL) was added and the reaction mixture was evaporated to dryness. The residue was dissolved in ethyl acetate (400 mL). The organic solution was quickly washed with 10% citric acid (200mL), H₂O (3X 100mL), saturated NaHCO₃(100mL) and brine (2X 100mL) over Na₂SO₄Drying and evaporating to dryness. The crude product was chromatographed on silica, eluting with 10% EtOAc/hexanes containing 0.2% TEA, and co-evaporated from toluene (100mL) and cyclohexane (2 × 100mL) to give the protected product 1b (P ═ Cbz, R ¹SMe) (7.2g, 54%) as a colorless oil:

¹HNMR(300MHz，CD₃SOCD₃)δ7.34-7.31(m，5)，7.05(d，J＝8.2，1)，5.19(d，J＝5.8，1)，5.01(d，J＝1.6，2)，3.99(apt dt，J＝5.5，9.3，9.3，2)，3.93-3.86(m，3)，3.49(dd，J＝2.5，9.6，1)，2.01(s，3)，1.03(d，J＝6.3，3)，0.10(s，9)，0.09(s，9)，0.04(m，18).

to a solution of dimethyl sulfoxide (413. mu.L, 5.82mmol) in DCM (1.5mL) cooled to-72 ℃ was added oxalyl chloride 2M solution in DCM (1.49mL, 2.98mmol) over 1 min. After 25min the protected product 1b (1.92g, 2.84mmol) was added via cannula. The resulting reaction mixture was stirred for 25min, then warmed to-50 ℃ (dry ice acetonitrile) and maintained at that temperature for 2 h. TEA (1.29mL, 3.30mmol) was added to the reaction mixture. After 25min, the reaction mixture was diluted with EtOAc (300 mL). The resulting organic solution was quickly washed with 5% citric acid (300mL), H₂O (2X 300mL), saturated NaHCO₃(100mL) and brine (100mL) over Na₂SO₄Dried and evaporated to dryness with toluene (100mL) to give the product 1 c. After co-evaporation with n-pentane and removal of residual solvent under high vacuum, the product 1c (P ═ Cbz, R) is obtained¹SMe) as a colorless crystalline solid (1.60g, 94%):

¹H NMR(300MHz，CDCl₃)δ7.37-7.33(m，5)，5.60(m，1)，5.21(d，J＝5.2，1)，5.17(d，J＝12.4，1)，5.08(d，J＝12.4，1)，4.74(m，1)，4.16-4.12(m，2)，3.87(d，J＝2.2，1)，3.69(dd，J＝2.5，9.3，1)，2.01(br s，3)，1.90(s，3)，0.19(s，9)，0.16(s，9)，0.15(s，9).

method B

Boc-protected product 1c (P ═ Boc, R)¹SMe) can generally be prepared as follows. To a suspension of 1a (MTL) (dried under high vacuum at 50 ℃ C.) (21.8g, 86mmol) in methanol (200mL) and TEA (26mL) cooled to 0 ℃ on ice was added di-tert-butyl dicarbonate (57.0g, 0.26 mol). The reaction mixture was then stirred at room temperature overnight. Toluene (100mL) was added to the reaction mixture and the solvent was removed to a total volume of 100mL, leaving a thick suspension to which cyclohexane (300mL) was added. The resulting solid precipitate was triturated, then filtered, washed with cyclohexane, ether and pentane and dried to constant weight. The crude Boc-protected product was used without further purification (87%): TLC R _f＝0.75(10％MeOH/DCM)；MS(ESPOS)：354[M+H]⁺；¹H NMR(300MHz，CD₃OD)δ0.14(d，J＝6.3，3)，1.43(s，9)，2.07(s，3)，3.55(dd，J＝3.3，10.43，1)，3.84-4.08(m，3)，4.10-4.15(m，2)，5.25(d，J＝5.5，1).

To a solution of N-Boc-1-methylthiolincosamide (240mg, 0.68mmol) in DMF (5mL) at 0 deg.C were added BSTFA (0.52mL, 2.0mmol) and triethylamine (0.14mL, 1.42mmol), followed by stirring at room temperature overnight. DMF was removed and the crude product was quickly passed through a silica gel column (pretreated with 2% triethylamine in ethyl acetate) and eluted with 10% ethyl acetate in hexane to give 1b (P ═ Boc, R)¹SMe) (350mg, 95%). To a solution of oxalyl chloride (0.16mL, 0.78mmol) in dichloromethane (5mL) was added dimethyl sulfoxide (0.22mL, 0.78mmol) slowly at-60 deg.C, then stirred for 15 min. A solution of 1b (370mg, 0.65mmol) in dichloromethane (5mL) was then added slowly. The reaction mixture was stirred for 45min, during which time the reaction temperature was increased to-40 ℃. Triethylamine (0.70mL, 3.25mmol) was then added and stirring continued at-40 ℃ for another 15 min. Then extracted with dichloromethane (100mL) and washed with 10% citric acid (50 mL). The residue obtained after removal of the solvent was then purified over a silica gel column using 10% ethyl acetate in hexane as eluent to give 1c (P ═ Boc, R)¹SMe) as a colourless oil (289mg, 78%): TLC: rf 0.60 (10% EtOAc/hexanes); ms (espos): 590[ M + Na ] ]+；1H NMR(300MHz，CDCl3)δ0.11(s，18)，0.17(s，18)，1.40(s，9)，1.84(s，3)，2.26(s，3)，3.63(dd，J＝2.7，9.34，1)，3.82(d，J＝1.9，1)，4.01-4.12(m，2)，5.15(d，J＝5.5，1).

Method C

In N₂Triphenylphosphonium bromide (3.29g, 9.20mmol) and potassium tert-butoxide (715mg, 6.4mmol) were suspended in toluene (31mL) under vigorous stirring. After 4.0h, the protected product 1c (P ═ Cbz, R) was added via a cannula¹SMe) (1.40g, 2.36mmol) in toluene (20 mL). The resulting reaction mixture was stirred for 2h, then diluted with EtOAc (250 mL). Subjecting the organic solution to rapid hydrogenation with H₂O (2X 100mL), brine (1X 100mL), Na₂SO₄Drying and evaporating to dryness. The crude product was chromatographed on silica, eluting with 6% EtOAc/hexanes containing 0.2% TEA, to give the ene product 2a (P ═ Cbz, R¹SMe，R²' -H) as a colorless oil, which crystallized upon co-evaporation from toluene and cyclohexane (0.65g, 46%):

¹H NMR(300MHz，CDCl₃)δ7.35-7.27(m，5)，6.36(d，J＝7.1，1)，5.24(d，J＝5.5，1)，5.08(m，4)，4.34(m，1)，4.16(m，2)，3.88(d，J＝2.2，1)，3.61(dd，J＝2.2，9.3，1)，2.20(s，3)，1.79(s，3)，0.17-0.13(m，27).

to 10% palladium on carbon (degusa wet, 50% w/w water) (700mg) in a Parr bottle was added the product 2a (P ═ Cbz, R)¹＝SMe，R²' ═ H) (490mg, 0.82mmol) in ethanol (50 mL). Using the bottle H₂Decontaminate and aerate to 65psi, shake for 24 h. The reaction mixture was filtered through celite, washing with methanol. Transferring the organic solution to a solution containing dried washed50w-400x H⁺Resin funnel of form (0.8g) was shaken for 10 min. After washing the resin three times with methanol and two times with water, and then with 5% TEA in MeOH (35 mL. times.10 min. times.5), the saturated product 2b was eluted from the resin. The combined filtrates were evaporated to dryness and coevaporated twice from EtOH, from 1: 1MeCN/H ₂Freeze-drying in O to obtain the product 2b (R)²' -H) as a colourless powder (198mg, 96%):

¹H NMR(300MHz，D₂O)δ5.17(d，J＝5.8，1)，3.97-3.84(m，3)，3.52(dd，J＝3.0，10.0，1)，2.82(dd，J＝4.4，8.5，1)，1.94(s，3)，1.89-1.81(m，1)，0.82(d，J＝6.9，3)，0.72(d，J＝6.9，3)；MS(ESPOS)：252.2[M+H]⁺，(ESNEG)：250.4[M-H]^-.

method D

In an alternative, when using the Boc protecting group in scheme 1 (P ═ Boc), methyltriphenylphosphonium bromide (12g, 33.6mmol) and potassium tert-butoxide (3g, 26.7mmol) were dissolved in THF (70mL) at 0 ℃ and stirred at rt for 4 h. Then the Boc-protected product 1c (P ═ Boc, R) was added¹SMe) (4.7g, 8.2mmol) in THF (30mL) and stirred at rt for 2 h. It was then extracted with EtOAc (300mL), washed with brine (100mL) and dried over sodium sulfate. Crude alkene product 2a (P ═ Boc, R¹＝SMe，R²' ═ H) was purified by silica gel column chromatography using 10% EtOAc in hexane as eluent (4.1g, 87.6%):

TLC：R_f0.5 (10% of EtOAc in hexanes):¹H NMR(300MHz，CD₃OD)δ7.24(m，2)，5.22(d，J＝5.7，1)，4.21(m，1)，4.09(m，2)，3.87(d，J＝2.4，1)，3.60(dd，J＝2.7，9.3，1)，1.99(s，3)，1.76(s，3)，1.43(s，9)；MS(ESPOS)：444[M-2TMS+Na]⁺.

to product 2a (P ═ Boc, R)¹＝SMe，R²' ═ H) in methanol (30mL) was addedH⁺Resin (1g), stirred at rt for 1 h. The resin was filtered, the solvent removed, and the resulting product (2.4g, 6.8mmol) dissolved in MeOH (30mL), Pd/C (2.5g) added and hydrogenated at 55psi overnight. After filtration and removal of solvent, the resulting crude product was purified by silica gel column chromatography eluting with 10% MeOH in DCM to give Boc-protected 7-methyl MTL as a white solid (2.06g, 86%): TLC R _f＝0.5(10％ofMeOH in DCM)；¹H NMR(300MHz，CD₃OD)δ5.23(d，J＝5.4，1)，4.11(m，1)，3.97(d，J＝10.2，1)，3.84(m，1)，3.52(m，1)，2.08(s，3)，1.44(s，9)，1.14(m，1)，0.93(d，J＝6.9，3)，0.85(d，J＝6.9，3)；MS(ESPOS)：351[M+H]⁺.

To a solution of Boc-protected 7-methyl MTL (150mg, 0.43mmol) in dichloroethane (6mL) was added dimethyl sulfide (0)16mL, 2.5mmol), followed by TFA (2mL), water (0.16mL) was stirred at rt for 1 h. The solvent is removed to give the crude product 2b (R)¹＝SMe，P＝Boc，R²' -H). Purification by column chromatography on silica gel using 30% MeOH in DCM as eluent gave product 2b (R)²' ═ H), identical in all respects to the product obtained in process C.

Method E

In N₂Sodium hydride (80mg, 3.3mmol) was suspended in THF (4mL) under an atmosphere while stirring vigorously. The suspension was cooled to-30 ℃ and diethyl (cyanomethyl) phosphonate (805. mu.L, 5.0mmol) was added. After 30min, the protected product 1c (P ═ Cbz, R) was added via a cannula¹SMe) (1.0g, 1.7mmol) in THF (3 mL). The resulting reaction mixture was stirred for 4h, then diluted with EtOAc (250 mL). The resulting organic solution was quickly taken up with saturated aqueous NaHCO₃(1X 100mL), brine (1X 50mL), and Na₂SO₄Drying and evaporating to dryness. The crude product was chromatographed on silica eluting with 6% EtOAc/hexanes to 10% EtOAc/hexanes containing 0.2% TEA to give the protected alkene product 2a (P ═ Cbz, R)¹＝SMe，R²' -CN) as a colorless oil (0.38g, 37%): ms (espos): 625.5[ M + H ]⁺，MS(ESNEG)：659.5[M+Cl]^-.

To 10% palladium on carbon (degusa wet, 50% w/w water) (300mg) in a Parr bottle was added the product 2a (P ═ Cbz, R)¹＝SMe，R²' -CN) (180mg, 0.29mmol) in ethanol (15mL) and concentrated HCl (29 μ L) was added. Using the bottle H₂Decontaminate and aerate to 65psi, shake for 24 h. The reaction mixture was filtered through celite, washing with methanol. Transferring the organic solution to a solution containing dried washed50w-400x H⁺Resin funnel of form (1g) shaken for 10 min. After washing the resin twice with methanol and with water, with 5% TEA in MeOH (20 mL. times.20 min. times.3) and with MeCN (20 mL. times.20 min), the resin was eluted with saturationProduct 2b (R)¹＝SMe，R²' -CN). The combined organic filtrates were evaporated to dryness from 1: 1MeCN/H₂Freeze-drying in O to obtain the product 2b (R)¹＝SMe，R²＝CH₂CN) as a colorless solid (70mg, 91%): ms (esneg): 275.3[ M-H]^-.

Method F

To the protected product 1c (P ═ Cbz, R) at 0 ℃¹SMe) (0.75g, 1.3mmol) in THF (7.3mL) was added MeMgCl 3m (aldrich) in THF (7.0mL, 2.1 mmol). The reaction mixture was heated to 4 ℃ over 3min and saturated with aqueous NH 1: 3 after 4h₄Cl/H₂The reaction mixture was quenched with O (10 mL). The quenched mixture was diluted to 100mL with water and extracted with DCM (4X 50 mL). The combined organic phases were dried and evaporated. The residue was dissolved in 1: 2: 4H ₂O/HOAc/THF (100mL), stirred for 20h, then evaporated with toluene (2X 100 mL). Chromatography (10: 1 to 10: 2DCM/MeOH) afforded the product 3a (P ═ Cbz, R)¹＝SMe，R²”＝Me)(153mg，31％)：MS(ESNEG)：399.5[M-H]^-.

To 10% palladium on carbon (degusa wet, 50% w/w water) (400mg) in a Parr bottle was added the product 3a (P ═ Cbz, R)¹＝SMe，R²"═ Me) (79mg, 0.2mmol) in ethanol (10 mL). Using the bottle H₂Decontaminate and aerate to 65psi, shake for 6 h. The reaction mixture was filtered through celite, washing with methanol. The combined filtrates were evaporated to dryness from 1: 1MeCN/H₂Freeze-drying in O to obtain the product 3b (R)¹＝SMe，R²"═ Me) as a colorless powder (42mg, 80%):

¹H NMR(300MHz，D₂O)δ5.33(d，J＝5.8，1)，4.83-4.06(m，3)，3.65-3.60(m，1)，3.06-3.03(m，1)，2.18(s，3)，1.30(s，3)，1.23(s，3)；MS(ESPOS)：268.4[M+H]，MS(ESNEG)：266.2[M-H]^-.

method G

To Boc-protected product 1c (P ═ Boc, R)¹＝SMe)(100mg, 0.18mmol) in methanol (3mL) O-trimethylsilylhydroxylamine (0.10mL, 0.88mmol) was added and stirred at rt overnight. Removal of the solvent gave the crude Boc-protected product 4a (R)¹＝SMe，R⁷H). To the crude product 4a (95mg, 0.15mmol) was added a solution of 30% trifluoroacetic acid in dichloroethane (10mL) and dimethylsulfide (0.5mL) and stirred for 1 h. The solvent is removed and the product 4b (R) is obtained¹＝SMe，R⁷H) was used directly in the next step.

TLC：R_f＝0.35(10％MeOH/DCM)；MS(ESPOS)：267(M+H)；¹H NMR(300MHz，CD₃OD)δ1.96(s，3)，2.09(s，3)，3.58(dd，J＝3.3，10.2，1)，3.90(s，1)，4.11(dd，J＝5.7，10.20，1)，4.19(d，J＝5.4，1)，4.50(d，J＝5.1，1)，5.36(d，J＝5.7，1).

Method H

To Boc-protected product 1c (P ═ Boc, R)¹To a solution of SMe) (100mg, 0.176mmol) in methanol (4mL) and water (1mL) was added O-alkylhydroxylamine hydrochloride (e.g. O-methylhydroxylamine hydrochloride) (60mg, 0.70mmol) and sodium acetate (57mg, 0.70mmol), heated at 80 ℃ for 3h and then stirred at rt overnight. The solvent was removed under high vacuum to give crude Boc-protected product 4a (R) ¹＝SMe，R⁷Me). The crude product 4a was dissolved in 30% trifluoroacetic acid in dichloroethane (10mL) and dimethylsulfide (0.5mL) and stirred at rt for 1 h. The solvent was removed and the residue was kept under high vacuum for 1h, product 4b (R)¹＝SMe，R⁷Me) was used directly in the next step: TLC Rf ═ 0.63 (10% MeOH/DCM); ms (espos): 281[ M + H]+；1H NMR(300MHz，CD3OD)δ1.95(s，3)，2.08(s，3)，3.60(dd，J＝3.3，10.2，1)，3.92(s，3)，4.13(dd，J＝4.8，10.2，1)，4.49(d，J＝1.2，1)，5.38(d，J＝5.4，1).

Method I

To Boc-protected product 1c (P ═ Boc, R)¹SMe) (500mg, 0.88mmol) in THF (10mL) tetrabutylammonium fluoride (2.5mmol, 1M THF solution) was added and the reaction mixture was stirred at rt for 1 h. Removing the solvent and passing the residue through siliconGel column purification using 5% methanol in dichloromethane as eluent. The product from the column (111mg, 0.31mmol) was then dissolved in a mixture of dichloromethane (3mL) and pyridine (3mL), to which was added acetic anhydride (0.5mL, 10.6mmol) and dimethylaminopyridine (80mg, 1.7mmol) and stirred at rt overnight. The solvent was removed and the crude product was purified over silica gel column using 30% ethyl acetate in hexane as eluent to give 5a (P ═ Boc, R)¹＝SMe)(58mg，3％)：

TLC R_f0.73 (50% EtOAc/hexanes);¹H NMR(300MHz，CDCl₃)δ1.38(s，9)，1.91(s，3)，1.98(s，3)，2.07(s，3)，2.18(s，3)，4.33(m，1)，4.72(m，1)，4.94(m，1)，5.21(m，2)，5.45(s，1)，5.57(m，1)；MS(ESPOS)：500[M+Na]⁺.

to product 5a (P ═ Boc, R)¹SMe) (158mg, 0.331mmol) in DCM (5mL) was added dimethylaminosulfur trifluoride (732 μ L, 3.31mmol) and stirred overnight. More DCM was added and the organic portion was washed with sodium bicarbonate. The residue obtained from removal of the solvent was purified by silica gel column chromatography using 20% ethyl acetate in hexane as eluent (100mg, 60%) to give the protected product (P ═ Boc, R) ¹SMe). The Boc-protected product was dissolved in 30% trifluoroacetic acid in dichloroethane and dimethylsulfide and stirred at rt for 1 h. Removal of the solvent gave the product 5b (R)¹＝SMe)：

TLC R_f0.63 (40% MeOH/hexane);¹H NMR(300MHz，CDCl₃)δ1.40(s，9)，1.69(t，J＝18.9，3)，1.98(s，3)，2.08(s，6)，2.13(s，3)，4.22-4.30(m，1)，4.53(dd，J＝10.9，25.3，1)，5.16-5.28(m，2)，5.52(s，1)，5.63(d，J＝5.2，1)；MS(ESPOS)：522[M+Na]⁺.

method J

Preparation of compound 6a (P ═ TFA)

To a 1L round bottom flask was added dry 1a MTL (R)¹SMe) (dried overnight under vacuum at 50 ℃) (20g,0.079mol), dry methanol (200mL), triethylamine (8.77g, 0.087mol) and methyl trifluoroacetate (127.3g, 0.99 mol). The reaction mixture was stirred at rt for 4h, then the solvent was evaporated to dryness to give the protected MTL 6a (R)¹SMe, P ═ TFA) (26.2g, 95%) was used directly in the next step.

Chloromethylenepiperidinium HCl (scheme 6, reagent b)

To a 3L three-necked round bottom flask equipped with a mechanical stirrer, glass stir bar (large Teflon paddle) was added diethyl ether (anhydrous, 1.8L) and N-formylpiperidine (35.6g, 0.315mol) under a nitrogen atmosphere. The reaction mixture was cooled to 0 ℃, triphosgene (31.2g, 0.105mol) was added in at least 5 portions over 2 hours, maintaining 0 ℃ and vigorous stirring. The reaction mixture was then warmed to rt (1hr) to ensure complete reaction of the triphosgene, and then cooled again to 0 ℃. The reaction mixture was then filtered under a stream of nitrogen or argon (very hygroscopic, malodorous) and washed with cold diethyl ether (2 × 100 mL). The resulting white crystals were then dried in vacuo to give chloromethylene piperidinium HCl (46.4g, 95%).

TFA protected 7-Cl MTL

To a 3L three-necked round bottom flask equipped with a mechanical stirrer, glass stir bar, Teflon paddles and reflux condenser was added chloromethylene piperidinium HCl (44.4g, 0.286mol) and dichloroethane (anhydrous, 1L) under a nitrogen atmosphere. The resulting slurry was stirred vigorously, and the temperature was adjusted to 0 ℃. Crude 6a (R) was added to the stirred reaction mixture over 1 minute¹SMe, P ═ TFA) (20g, 0.057 mol). The reaction mixture was stirred for 1hr, then the temperature was raised to 65 ℃ during which time the reaction was seen to become a clear solution. The reaction mixture was then stirred at 65 ℃ for 18 h. The reaction mixture was then cooled to 0 ℃ and then poured quickly into a 4L Ellemaie bottle equipped with a mechanical stirrer, to which water (1L) and NaOH (22.9g, 0.57mol) had been added and cooled to 0 ℃. The reaction mixture was then stirred for 30min and then the pH was adjusted to 10.5(pH paper) with concentrated HCl (over 5min addition while checking the pH after each HCl addition and simply adding if the pH drops below 10.5NaOH adjusted to 10.5 is acceptable). This pH adjusted mixture was then stirred for 2 hours while allowing the reaction to return to rt. The pH was then adjusted to 7 with more concentrated HCl and then stirred overnight, or until the product was seen to be free of adducts formed by the chlorinating agent and the OH functional groups of the sugars. The reaction mixture is then evaporated to dryness by means of a high vacuum attached to a rotary evaporator (co-evaporation with a solvent, such as toluene, may be employed to facilitate this process). The resulting solid was then charged with a 10% methanol/DCM mixture and stirred for 1 hour to make the product salt free. The mixture was then filtered and the filtrate evaporated to dryness to give a syrup containing the product and N-formylpiperidine. The mixture was triturated with hexane, which was decanted several times from the product oil to remove most of the N-formylpiperidine. The crude reaction product was then chromatographed, eluting with 10% methanol in DCM, to give purified TFA-protected 7-ClMTL (21.1g, 75%).

6b(R¹＝SMe，R²＝Cl，R³＝H).

A500 mL three-necked round bottom flask equipped with a mechanical stirrer was charged with a solution of purified TFA-protected 7-Cl MTL (20g, 0.054mol) in a small amount of methanol (10mL) followed by 1M NaOH (250mL) at 0 ℃. The reaction mixture was then stirred at 0 ℃ for 12hr (first see the mixture to form a very viscous solid, eventually going into solution, soon see pure product crystals formed), while periodically harvesting pure product crystals to prevent hydrolysis of the 7-chloro functionality, washing with a small amount of cold water, followed by cold methanol, to give 7-Cl MTL 6b (R)¹＝SMe，R²＝Cl，R³H) as a colorless solid (10g, 50%).

Method K

According to Zhang, r.; et al, Journal of the American chemical society, 1998, 120, 3894-3902, enolization (LiHMDS) and alkylation of 7a with 4-bromo-2-methyl-2-butene to give lactam 7b (R)⁹' -2-methyl-2-butene) diastereomer mixture (61%). Compound 7a is available from commercial suppliers, such as Bachem. Alternatively, 7a may be assisted by the bookMethods well known in the art, see, for example, Baldwin j.e.; et al.; tetrahedron, 1989, 45, 7449-.

Reduction of lactam 7b to pyrrolidine 7c (R) by a two-step sequence ⁹' -2-methyl-2-butene) (70%) which involves conversion of the lactam to the hemiaminalReduction and subsequent conversion of the hemiaminal by Et₃SiH/BF₃·OEt₂And (4) reducing. Pyrrolidine 7c (778mg, 2.08mmol), 10% palladium on carbon (230mg) was subjected to a 50psi Parr hydrogenolysis in dry methanol (25mL) for 5 h. The reaction mixture was filtered through a pad of celite, washing several times with methanol. The combined washings and filtrate were evaporated to dryness to give colorless oil 7d (R)⁹"═ 2-methyl-2-butane): TLC R_f0.3[ solvent system: DCM/hexane/MeOH (6: 5: 1)]；MS(ESNEG)：284.5[M-H]^-.

Method L

At-78 ℃ and N₂Next, to the stirred solution of 7a (9.47g, 29.7mmol, 1equiv) in anhydrous THF was added a 1M solution of LiHMDS in THF (33mmol, 33mL, 1.1equiv) followed by cis-1-bromo-2-pentene (4.21mL, 35.6mmol, 1.2equiv) to afford lactam 7b (R) after purification on silica gel⁹' -2-pentene) (43.2%). Reduction of lactam 7b (3.96g, 10.22mmol) to pyrrolidine 7c (R) by a two-step sequence⁹' -pentene) which involves lactam to hemiaminal in anhydrous THF at-78 ℃Reduction, and subsequent Et-78 deg.C of hemiaminal in anhydrous DCM₃SiH/BF₃·OEt₂Reduction to give 7c (R) after purification on silica gel⁹' -2-pentene) (71%). Pyrrolidine 7c (2.71g, 7.26mmol) and 10% palladium on carbon (560mg) were subjected to a 50psi Parr hydrogenolysis in dry methanol (30mL) for 5 h. The reaction mixture was filtered through a pad of celite, washing several times with methanol Next, the process is carried out. The combined washings and filtrate were evaporated to dryness to give colorless oil 7d (R)⁹Pentyl) (1.68g, 80%): TLC: r_f0.3[ solvent system: DCM: hexane: MeOH (6: 5: 1)].MS(ESNEG)：284.5[M-H]^-.

Method M

8a(R⁹' 3, 3-difluoropropan-2-ene)

7c(R⁹' -2-methyl-2-butene) ozonolysis treatment in anhydrous dichloromethane followed by treatment with DMS at-78 ℃ followed by slow warming to rt afforded the terminal aldehyde 8a (77%) which was used in the next step without further purification.

To a solution of aldehyde 8a (407mg, 1.17mmol, 1equiv) obtained from the above reaction in dimethylacetamide (0.25mL) was added dibromodifluoromethane (0.21mL, 2.34mmol, 2equiv) at 0 ℃. To the stirred mixture was added a solution of triphenylphosphine (0.61g, 2.34mmol, 2equiv) in dimethylacetamide (0.5mL) over 20 minutes under nitrogen. The reaction mixture was warmed to rt, stirred for 30 min, and then added to activated zinc (0.25g, 3.82mmol, 3.3equiv) via dimethylacetamide (0.3 mL). The resulting reaction mixture was stirred at 110 ℃ for 1h, cooled to rt, and filtered with dimethylacetamide (7 mL). The filtrate was poured into ice water (100mL) and extracted with ether (150 mL). The ether layer was washed with brine, dried and concentrated. The residue was chromatographed to give clear oil 8a (R) ⁹' -3, 3-difluoroprop-2-ene) (182mg, 41%): ms (espos): 282.4[ M Boc + H]⁺.

8c(R⁹' -3, 3-difluoropropan-2-ene to a solution of 8a (84.1mg, 0.22mmol, 1equiv) in THF (3mL) and water (1mL) was added lithium hydroxide monohydrate (46.3mg, 1.10mmol, 5 equiv). The reaction mixture was stirred at rt overnight. THF was removed under vacuum. The residue was dissolved in ethyl acetate (50mL) and partitioned with 10% citric acid (20 mL). The organic layer was washed with water (1x), brine (1x), dried, and concentrated to give 8c (R)⁹' -3, 3-difluoroprop-2-ene) as a clear glass (56mg, 87%): ms (espos):192.3[M-Boc+H]⁺；MS(ESNEG)：290.3[M-H]^-.

8b(R⁹"═ 3, 3-difluoropropane". the saturated product of scheme 8 can be obtained by means of a hydrogenation process, as described, for example, for process K of 7 d.

Method N

9b(P＝Boc，m＝1，LG＝Ts).

To a solution of N-Boc- (2S, 4R) -4-hydroxyproline methyl ester (Bachem)9a (P ═ Boc, m ═ 1) (5g, 20.4mmol, 1equiv) and DMAP (0.25g, 2.04mmol, 0.1equiv) in DCM (80mL) was added toluenesulfonic anhydride (8.65g, 26.5mmol, 1.3 equiv). The reaction mixture was cooled to 0 ℃ and pyridine (6.59mL, 81.5mmol, 4equiv) was added. The mixture was stirred at 0 ℃ for 30 minutes and then at rt overnight. The solution was concentrated to dryness. The residue was dissolved in ethyl acetate (400mL) and treated with 10% aq. citric acid (2X 400mL), sat ₃(400mL) and brine, over Na₂SO₄Dried and concentrated to give yellow syrup 9b (P ═ Boc, m ═ 1, LG ═ Ts) (8.44g, 100%): HPLC (method RV-1), C183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 min; 1.5 mL/min): rt 3.096.

9c(P＝Boc，m＝1，R⁹2, 4-dichlorobenzylthio).

In N₂Next, 2, 4-dichlorobenzylthiol (1.48g, 7.66mmol, 3equiv) was added to a solution of tosylate 9b (P ═ Boc, m ═ 1, LG ═ Ts) (1.02g, 2.55mmol, 1equiv) in anhydrous DMF (7.6mL), followed by MTBU (0.55mL, 3.83mmol, 1.5 equiv). The reaction mixture was stirred at rt overnight and concentrated to dryness. The residue was dissolved in ethyl acetate (100mL), washed with 10% citric acid (50mL) and brine, and concentrated. The residue was chromatographed to give clear syrup 9c (P ═ Boc, m ═ 1, R⁹4- (2, 4-dichlorobenzylthio) (1.0 g): ms (espos): 320.2[ M-Boc + H]⁺，MS(ESNEG)：418.4[M-H]^-.

To methyl ester 9c (P ═ Boc, m ═ 1, R⁹＝2，4-Dichlorobenzylthio) (1.0g, 2.38mmol, 1equiv) in THF (9mL) and water (3mL) was added lithium hydroxide (0.5g, 11.9mmol, 5 equiv). The reaction mixture was stirred at rt overnight. THF was removed under vacuum. The residue was partitioned between ethyl acetate (150mL) and 10% citric acid (100 mL). The organic layer was washed with water (1X), brine (1X) and over Na ₂SO₄Drying and evaporation gave clear syrup 9d (P ═ Boc, m ═ 1, R)⁹2, 4-dichlorobenzylthio) (1.0 g): ms (espos): 306.3[ M-Boc + H]⁺；MS(ESNEG)：404.2[M-H]^-.

Process O

4-Propylpyridine-2-carboxylic acid 10b (R)⁹N-propyl).

To 4-propylpyridine (TCI) (2.5g, 20mmol) was added 30% hydrogen peroxide (2.4g) and refluxed overnight. The solvent was removed and the resulting residue was dissolved in DCM (30 mL). Trimethylsilylcyanide (2.6g, 26mmol) was added to the above solution followed by dimethylcarbamoyl chloride (2.8g, 26mmol) and stirred at room temperature overnight. Potassium carbonate (10%, 100mL) was added. The organic layer was separated, dried over sodium sulfate, and then concentrated to give 4-propyl-2-cyanopyridine (2.5g, 93%). Then refluxed overnight in hydrochloric acid (6N, 60 mL). Crystallization from acetonitrile yields 4-propylpicolinic acid 10b (R)⁹＝nPr)(2.0g，71％)：MS(ESPOS)：166[M+H]；¹HNMR(300MHz，CD₃OD)δ8.75(dd，J＝9.0，3.0，1)，8.42(s，1)，8.08(dd，J＝9.0，3.0，1)，3.00(t，J＝7.5，2)，1.82(m，2)，1.05(t，J＝7.2，3).

4-propyl- (3-phenyl) pyridine-2-carboxylic acid 10b (R)⁹4-propyl- (3-phenyl)).

Trimethylsilylcyanide (1.3mL, 10mmol) and dimethylcarbamoyl chloride (1mL, 10mmol) were added to a solution of 4-propyl- (3-phenyl) pyridine-N-oxide (1g, 4.69mmol) in dichloromethane (10mL) and stirred at room temperature for 24 hours. Aqueous potassium carbonate (10%, 10mL) was added and extracted with dichloromethane (100 mL). The crude product obtained by removing the solvent was dissolved in hydrochloric acid (6N, 30mL) and refluxed for 24 hours. Removing the acid, followed by subjecting the crude product to Crystallization from acetonitrile gave acid 10b (1g, 86%): ms (espos): 240[ M-1 ]]；¹H NMR(300MHz，CD₃OD)δ2.03-2.17(m，2)，2.74(t，J＝7.2，2)，3.04(t，J＝7.8，2)，7.16-7.38(m，5)，8.07(d，J＝4.2，1)，8.40(s，1)，8.71(d，J＝5.7，1).

Method P

4-Chloropyridinecarboxylic acid methyl ester

A mixture of picolinic acid (20g, 162mmol, 1equiv) and sodium bromide (33.43g, 325mmol, 2equiv) in thionyl chloride (81mL) was refluxed for 5 h. The solvent was removed under vacuum. Anhydrous methanol (160mL) was added and the mixture was stirred at rt for 30 min. The solvent was evaporated, the residue was dissolved in 5% sodium bicarbonate and extracted with ethyl acetate (3 ×). The organic layers were combined and MgSO₄Drying and evaporating. The residue was purified by chromatography to give methyl 4-chloropicolinate (19.9g, 72%) as a white solid:¹HNMR(300MHz，CDCl₃)δ8.63(d，J＝5.4，1)，8.13(d，J＝2.1，1)，7.48(dd，J＝2.0，5.3，1)，4.00(s，3).

4-iodopicolinic acid 11a.

A mixture of methyl 4-chloropicolinate (2.4g, 14.1mmol), 57% hydroiodic acid (13.3mL), and 50% aqueous hypophosphorous acid (0.66mL) was stirred at 85 ℃ for 2h, then at 107 ℃ overnight. The mixture was cooled to 95 ℃. At this temperature, 10M aqueous sodium hydroxide (4.2mL) was added over 30 minutes, followed by water (15.2 mL). The mixture was cooled to rt and stirred at rt for 1 h. The precipitate was filtered, washed with cold water and dried under high vacuum overnight to give 11a as a yellow solid, 4-iodopicolinic acid (3.5g, 66%): ¹H NMR(300MHz，DMSO d₆)δ8.39(d，J＝5.1，1)，8.35(d，J＝1.8，1)，8.07(dd，J＝1.7，5.2，1)；MS(ESPOS)：250.2[M+H]⁺.

4-Iodopicolinic acid methyl ester 11b.

To a solution of 4-iodopicolinic acid 11a (7.0g, 18.6mmol) in MeOH (70mL) at 23 deg.C was added concentrated sulfurAcid (350. mu.L) and the reaction mixture was refluxed for 48 h. The reaction mixture was cooled to room temperature and concentrated to give the desired product methyl 4-iodopicolinate 11b (4.4g, 90%) as a yellow oil:¹H NMR(300MHz，CDCl₃)δ8.52(t，J＝0.6，1.5，1)，8.40(d，J＝5.1，1)，7.86-7.88(dd，J＝0.6，5.1，1)，4.02(s，3)；MS(ESPOS)：263.9[M+H]；285.9[M+Na].

4- [3- (tert-butyl-dimethyl-silanyloxy) -prop-1-ynyl]-pyridine-2-carboxylic acid methyl ester 11c (R)⁹' -tert-butyl-dimethyl-silanyloxy).

To the dried flask was added 11b (5.41g, 20.6mmol, 1equiv), triphenylphosphine (431.5mg, 1.65mmol, 0.08equiv), copper (I) iodide (313.4mg, 1.65mmol, 0.08equiv), palladium acetate (5mg, 0.82mmol, 0.04equiv) and triethylamine (74 mL). The mixture was degassed with nitrogen, followed by the addition of tert-butyldimethyl (2-propynyloxy) silane (Aldrich) (8.34mL, 41.14mmol, 2 equiv). The mixture was stirred at rt for 3 h. The solvent was removed under vacuum to give a dark residue. The residue was purified by chromatography to give the ester 11c (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -prop-1-ynyl) (6.07g, 97%) as a brown oil:¹HNMR(300MHz，CDCl₃)δ8.67(dd，J＝0.8，5.0，1)，8.09(m，1)，7.43(dd，J＝1.7，5.0，1)，4.54(s，2)，3.99(s，3)，0.92(s，9)，0.14(s，6).MS(ESPOS)：306.5[M+H]+.

4- [3- (tert-butyl-dimethyl-silanyloxy) -propyl ]Piperidine-2-carboxylic acid methyl ester 11d (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl).

To 11c (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -prop-1-ynyl) (6.05g, 19.8mmol, 1equiv) in a mixture of MeOH (60mL), water (60mL) and acetic acid (1.14mL, 19.8mmol, 1equiv) platinum oxide (2.0g) was added. The mixture was purged with hydrogen and aerated (50psi) and shaken overnight at rt. The platinum oxide was removed by filtration and the filtrate was concentrated to give the product 11d (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl) (5.0g, 80%): MS (ESPOS))：316.6[M+H]⁺.

4- [3- (tert-butyl-dimethyl-silanyloxy) -propyl]1-tert-butyl 2-methyl piperidine-1, 2-dicarboxylate 11e (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl, P ═ Boc).

To 11d (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl) (4.99g, 15.8mmol, 1equiv) in methanol (60mL) was added triethylamine (4.42mL, 31.7mmol, 2equiv) and di-tert-butyl dicarbonate (4.7mL, 20.6mmol, 1.3 equiv). The mixture was stirred at rt overnight. The solvent was removed under vacuum. The residue was purified by chromatography to give the carbamate 11e (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl, P ═ Boc) (2.75g, 42%) as a clear syrup:

¹H NMR(300MHz，CDCl₃)δ4.28(t，J＝6.6，1)，3.70(s，3)，3.55(t，J＝6.3，2)，3.55-3.48(m，1)，3.40-3.30(m，1)，2.00-1.92(m，1)，1.82-1.69(m，2)，1.64-1.20(m，6)，1.41(s，9)，0.86(s，9)，0.01(s，6)；MS(ESPOS)：316.6[M+H-Boc]+.

4- [3- (tert-butyl-dimethyl-silanyloxy) -propyl]1-tert-butyl (R) -piperidine-1, 2-dicarboxylate 11f⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl, P ═ Boc).

To 11e (R)⁹A mixture of 3- (tert-butyl-dimethyl-silanyloxy) -propyl, P ═ Boc) (2.75g, 6.63mmol, 1equiv) in THF (12mL) and water (4mL) was added lithium hydroxide monohydrate (306mg, 7.29mmol, 1.1 equiv). The mixture was stirred at rt overnight. Additional lithium hydroxide monohydrate (834mg, 19.89mmol, 3equiv) was added and the mixture was stirred at rt for 5 hr. THF was removed under vacuum. The aqueous layer was dissolved in ethyl acetate and partitioned with 10% citric acid. The organic layer was washed with water (1x), brine (1x), dried, concentrated to give a yellow syrup which was purified by chromatography to give the desired acid 11f (R)⁹' -3- (tert-butyl-dimethyl-silanyloxy) -propyl, P ═ Boc) (1.83g, 69%) as a colorless syrup:¹HNMR(300MHz，CDCl₃)δ4.26(t，J＝6.9，1)，3.57(t，J＝6.5，2)，3.53-3.44(m，1)，3.43-3.33(m，1)，2.05-1.96(m，1)，1.82-1.68(m，2)，1.64-1.45(m，3)，1.42(s，9)，1.37-1.27(m，3)，0.86(s，9)，0.02(s，6).MS(ESPOS)：424.7[M+Na]+.

method Q

(2S, 4R) -N-Boc-4-hydroxyproline methyl ester.

To a stirred solution of (2S, 4R) -4-hydroxyproline (Bachem) (25g, 108mmol) in methanol (50mL) at 0 deg.C was added trimethylsilyldiazomethane (24.6g, 216 mmol). The mixture was stirred at 0 ℃ for 1 h. The residue obtained from the removal of the solvent was purified by column chromatography, eluting with 50% ethyl acetate in hexane to give (2S, 4R) -N-Boc-4-hydroxyproline methyl ester (27g, 100%) as a colorless oil: ¹H NMR(300MHz，CDCl₃)δ4.47(m，1)，4.39(m，1)，3.70(s，3)，3.60(m，2)，2.27(m，1)，2.05(m，1)，1.38(s，9)；MS(ESPOS)：268(M+Na).

(2S, 4R) -N-Boc-4-ketoproline methyl ester 12a (P ═ Boc, P)₂＝Me，m＝1).

To a solution of oxalyl chloride (15g, 118mmol) in DCM (15mL) was slowly added DMSO (18.6mL, 236mmol) at-78 deg.C over 15 minutes. After the addition was complete, a solution of (2S, 4R) -N-Boc-4-hydroxyproline methyl ester (26.5g, 108mmol) in DCM (100mL) was added dropwise, stirred at-78 deg.C for 20min, then triethylamine (54.6g, 540mmol) was added and stirred for 2 h. The reaction mixture was then washed with 10% aq. HCl (200 mL). The organic layer was separated and dried over sodium sulfate. The crude product from solvent removal was purified by silica gel column chromatography eluting with 50% EtOAc in hexanes to give 12a (P ═ Boc, P)₂Me, m-1) (20g, 78%) as a brown solid:¹H NMR(300MHz，CDCl₃)δ4.80(m，1)，3.88(d，J＝8.7，2)，3.77(s，3)，2.98(m，1)，2.58(m，1)，1.45(s，9)；MS(ESPOS)：244(M+H).

N-Boc-4-hydroxy-4-allylproline methylEster 12b (P ═ Boc, P)₂＝Me，m＝1，R⁹' -allyl).

To stirred 12a (P ═ Boc, P)₂Me) (1g, 4.11mmol) in THF (10mL) was added tetraallyltin (1.08mL, 4.52mmol) in anhydrous THF, then cooled to 0 ℃, then boron trifluoride etherate (0.520mL, 4.11mmol) was added slowly. The mixture was stirred at 0 ℃ for 1h, then at room temperature for an additional 2 h. Potassium fluoride (360mg in 5mL of water) and celite (1g) were added and the reaction mixture was stirred for 1 hour. The reaction mixture was filtered and concentrated to dryness. The residue was dissolved in DCM (200mL), washed with water (100mL) and brine (100mL), over MgSO ₄Drying and evaporating to dryness. The resulting residue was purified by silica gel column chromatography eluting with 50% EtOAc in hexanes to give 12b (P ═ Boc, P)₂＝Me，m＝1，R⁹Allyl) (0.94g, 80%) as a colorless oil:

¹H NMR(300MHz，CDCl₃)δ5.87(m，1)，5.19(m，2)，4.34(m，1)，3.75(d，J＝4.8，3)，3.50(m，3)，2.37(m，1)，2.21(m，1)，1.39(d，J＝12.9，9)；MS(ESPOS)：308[M+Na]+.

N-Boc-4-fluoro-4-allylproline methyl ester 12c (P ═ Boc, P)₂＝Me，m＝1，R⁹' -allyl).

To a stirred solution of DAST (1.06g, 6.58mmol) in DCM (10mL) was slowly added 12b (P ═ Boc, P) at-78 ℃₂＝Me，R⁹Allyl) (940mg, 3.3mmol) in anhydrous DCM (10 mL). The mixture was then stirred at-78 ℃ for 1h, then at-10 ℃ for another 1 h. DCM (50mL) was added, followed by NH₄Cl (10%, 150mL) was quenched, the organic layer was separated, dried over sodium sulfate and evaporated to dryness. The residue was purified by silica gel column chromatography using 5% EtOAc in hexanes as the eluent to give the desired product 12c (P ═ Boc, P)₂＝Me，m＝1，R⁹Allyl) (330mg, 34%) as a colorless oil:

¹HNMR(300MHz，CDC₁₃)δ5.82(m，1)，5.12(m，2)，4.43(m，1)，3.66(s，3)，3.47(m，1)，2.37(m，1)，2.43(m，4)，1.37(dd，J＝4.5，13.8，9)；MS(ESPOS)：310[M+Na]+.

N-Boc-4-fluoro-4-propylproline methyl ester 12c (P ═ Boc, P)₂＝Me，m＝1，R⁹Propyl).

To 12c (P ═ Boc, P)₂＝Me，m＝1，R⁹Allyl) (0.33g, 1.15mmol) in MeOH (15mL) was added 10% Pd/C (40 mg). The reaction mixture was stirred at room temperature under hydrogen (30atm) for 3 hr. The catalyst was filtered through celite and washed with methanol. The filtrate was concentrated to give the desired protected amino acid ester 12c (P ═ Boc, P) ₂＝Me，R⁹Propyl) (0.33g, 100%) as a clear oil:¹HNMR(300MHz，CDCl3)δ4.43(m，1)，3.71(m，4)，3.47(m，1)，2.51(m，1)，1.98(m，5)，1.40(dd，J＝5.1，13.8，9)，0.93(J＝7.8，3)；MS(ESPOS)：190[M-Boc]+.

N-Boc-4-fluoro-4-propylproline 12d (P ═ Boc, R)⁹Propyl, m 1).

To a solution of methyl ester 12d (330mg, 1.15mmol) in THF (12mL) and water (4mL) was added lithium hydroxide monohydrate (60mg, 1.38 mmol). The reaction mixture was stirred at room temperature overnight. THF was removed and the residue was dissolved in ethyl acetate (50mL) and washed with 10% citric acid (100mL) and brine (20 mL). The organic portion was concentrated to give the desired protected amino acid 12e (P ═ Boc, R)⁹Propyl, m ═ 1) (310mg, 100%) as a white solid:

¹H NMR(300MHz，CD₃OD)δ4.43(m，1)，3.71(m，6)，2.51(m，2)，1.98(m，3)，1.45(m，9)，0.96(m，3)；MS(ESNEG)：274[M-1]

process R

(2S, 4R) -N-trifluoroacetyl-4-tert-butoxyproline.

To a mixture of 4-tert-butoxyproline (Bachem) (5.0g, 27mmol, 1equiv) and TEA (11.2mL, 80mmol,3equiv) in dry MeOH (30mL) was added ethyl trifluoroacetate (4.8mL, 40mmol, 1.5 equiv). The mixture was stirred at 24 ℃ overnight. The solution was concentrated to dryness, dissolved in DCM (200mL) and the organic phase was washed with aq.0.2M KHSO₄(2X 100mL) and brine (1X 100mL) over MgSO₄Drying and evaporating to dryness. The resulting residue was triturated with cyclohexane and pentane to give the product (2S, 4R) -N-trifluoroacetyl-4-tert-butoxyproline as a pale yellow powder (5.5g, 72%).

14a(P＝CF₃CO，m＝1，R²＝H，R³＝OH).

To a solution of MTL 1a (1.32g, 5.3mmol, 1equiv) in anhydrous DMF (16mL) at 0 deg.C was added triethylamine (2.20mL, 15.9mmol, 3equiv), followed by bis- (trimethylsilyl) trifluoroacetamide (2.81mL, 10.6mmol, 2.0 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. To the reaction mixture was added (2S, 4R) -N-trifluoroacetyl-4-tert-butoxyproline (1.8g, 6.3mmol, 1.2equiv), HATU (3.02g, 8.0mmol, 1.5 equiv). The reaction mixture was stirred for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate (500mL), and washed with 10% citric acid (100mL), water (100mL), half-saturated aqueous NaHCO₃(200mL) and brine wash. Subjecting the organic layer to Na₂SO₄Dried and evaporated to give a yellow syrup, which is dissolved in MeOH (100 mL). Adding driedH + resin (500mg), the resulting suspension was stirred for 50min, filtered and evaporated to dryness to give a yellow solid (2.89 g). The product was purified by silica chromatography eluting with DCM/hexane/MeOH 6: 5: 1 to 7: 2: 1 to give 14a (P ═ CF)₃CO，m＝1，R²＝H，R³OH) as a colorless solid (1.7g, 51%).

14b(P＝CF₃CO，m＝1，R²＝H，R³＝OAc).

At 0 deg.C, to 14a (P ═ CF)₃CO，m＝1，R²＝H，R³Acetic anhydride (3mL, 31mmol) was added to a solution of OH) (1.63g, 3.1mmol), pyridine (3mL, 30mmol), DMAP (38mg, 0.31mmol) in anhydrous DCM (10 mL). The reaction temperature was raised to 24 ℃ over 1h and stirred for 48 h. The reaction mixture was diluted with chloroform (200mL) and the organic phase was diluted with Aq.10% acetic acid (3X 200mL), 10% citric acid (200mL), half saturated aq ₃(200mL) and brine (1X 100mL) over Na₂SO₄Drying and evaporation gave the peracylated intermediate 14b (P ═ CF)₃CO，m＝1，R²＝H，R³OAc) (2.14g, 99%) as colorless crystals.

To the above-mentioned peracylated intermediate 14b (P ═ CF)₃CO，m＝1，R²＝H，R³To a solution of 2.1g, 3.1mmol) of DCE (64mL) and methyl sulfide (1.4mL) was added trifluoroacetic acid (21mL) and water (1.4 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with DCE. The residue was purified by chromatography eluting with 5% MeOH in DCM to give the intermediate alcohol (1.6g, 83%) as a colourless solid, which was used in the next step without characterisation.

14b(P＝CF₃CO，m＝1，R²＝H，R³＝OAc).

To a solution of the above 4-ol intermediate (1.5g, 2.38mmol, 1equiv) and DMAP (29mg) in DCE (9.5mL) was added p-toluenesulfonic anhydride (1.01g, 3.09mmol, 1.3 equiv). The reaction mixture was cooled to 0 ℃ and pyridine (0.77mL, 9.52mmol, 4equiv) was added. The reaction mixture was stirred at 0 ℃ for 30 minutes and then at rt overnight. The reaction mixture was concentrated to dryness. The residue was dissolved in ethyl acetate (200mL) and washed with 10% citric acid (2X 200mL), sat₃(200mL) and brine, over Na₂SO₄Drying and concentration gave a yellow syrup which was purified by chromatography eluting with 4: 1 hexanes/EtOAc to give the P-toluenesulfonate product 14b (P ═ CF) ₃CO，m＝1，R²＝H，R³OAc) (1.7g, 92%) as a colorless solid.

Method S

N-propyl malonic acid diethyl ester

To a suspension of sodium hydride (60% dispersion in mineral oil, 12.6g, 315mmol, 1.05equiv) in DMF (300mL) was added a solution of diethyl malonate (45.5mL, 300mmol, 1equiv) in DMF (100mL) via cannula at 23 ℃ over 10 min. The addition resulted in a slight exotherm, and H was observed₂The gas is released without cooling. After addition, the reaction was stirred at 23 ℃ for 45min, then treated with 1-bromopropane (27.3mL, 300mmol, 1 equiv). The reaction was stirred at 23 ℃ for 25min, then heated to 65 ℃ for 3h, then stirred at 23 ℃ overnight. The reaction mixture was added to 1.0N HCl (1N) and then extracted with diethyl ether (700 mL). Subjecting the ether extract to H₂O (400mL), brine (200mL), and dried (MgSO)₄) Filtered and concentrated to give 65.1g of product as a clear oil.¹³C NMR revealed a mono: di-alkylated product of about 4: 1. The product diethyl n-propylmalonate was used without further purification:

¹H NMR(300MHz，CDCl₃)4.18(q，J＝6.9Hz，4H)，3.32(t，J＝7.8Hz，1H)，1.91-1.79(m，2H)，1.41-1.25(m，2H)，1.25(t，J＝6.9Hz，6H)，0.92(t，J＝7.5Hz，3H)；¹³CNMR(300MHz，CDCl₃)

(. represents the signal due to the double alkylated secondary product)

61.2，60.9*，51.8，34.4*，30.7，20.5，17.3*，14.4*，14.0，13.7.

Ethyl n-propylmalonate 21b (R)⁹N-propyl).

To a solution of n-propylmalonic acid diethyl ester (contaminated with approximately 20% bis (n-propyl) malonate, 65.0g, 273mmol, 1equiv) in EtOH (500mL) at 23 ℃ was added 1.0m koh solution (273mL, 273mmol, 1 equiv). After addition, the reaction was heated to 80 ℃ (internal temperature) for 4 h. After cooling to 23 ℃, EtOH was removed in vacuo. The residual mixture was reacted with H in diethyl ether (400mL) ₂Partition between O (200 mL). Separating the layers, ether layerWith saturated aqueous NaHCO₃(100mL) was extracted. Combining aqueous NaHCO₃Layers were combined with the original aqueous layer, the solution was acidified to pH 1 with 1.0N HCl and extracted with EtOAc (2X 600 mL). The EtOAc extract was dried (MgSO)₄) Filtering, and concentrating to obtain 21b (R)⁹N-propyl) 41.3g (237mmol, 79%, 2 steps) of pure product as clear oil:

¹HNMR(300MHz，CDCl3)4.43(q，J＝7.2Hz，2H)，3.60(t，J＝7.5Hz，1H)，2.18-2.02(m，2)，1.66-1.50(m，2H)，1.49(t，J＝7.2Hz，3H)，1.15(t，J＝7.5Hz，3H).

n-propylacrylic acid ethyl ester 21c (R)⁹N-propyl).

To a solution of ethyl n-propylmalonate (41.3g, 237mmol, 1equiv) in EtOH (500mL) at 23 ℃ was added piperidine (28.1mL, 284mmol, 1.2equiv) followed by aqueous formaldehyde (37%, 88 mL). After addition, the reaction was refluxed for 29 h. After cooling to 23 ℃, the mixture was partitioned between diethyl ether (500mL) and 1.0N HCl (800 mL). The layers were separated and the aqueous layer was extracted with diethyl ether (500 mL). Combining the organic layers with H₂O (500mL), brine (300mL), and dried (MgSO)₄) Filtered and concentrated (rotary evaporation only, product is potentially volatile). The product was distilled under vacuum (bp 70 ℃ C., 15mmHg) to give 16.4g of 21c (R)⁹N-propyl) (115mmol, 49%) desired product:¹h NMR showed contamination by unidentified material; the product was used without further purification in the next step: ¹H NMR(300MHz，CDCl₃)6.12(s，1H)，5.50(s，1H)，4.19(q，J＝7.2Hz，2H)，2.23(t，J＝7.5Hz，2H)，1.55-1.42(m，2H)，1.24(t，J＝7.2Hz，3H)，0.92(t，J＝7.5Hz，3H).

2-propyl-prop-2-en-1-ol.

At 78 deg.C, over 20min through cannula to 21c (R)⁹N-propyl) (16.4g, 115mmol, 1equiv) CH₂Cl₂(500mL) solution DIBALH (1.0M in hexane, 403mL, 403mmol, 3.5equiv) was added. After addition, the reaction was stirred at 78 ℃ for 30min and then over a period of timeThe temperature was raised to 55 ℃ over 60 min. Once the reaction bath temperature had reached 55 ℃, EtOAc (15mL) was added to quench excess DIBALH. After stirring for 5min, the quenched reaction mixture was slowly added via cannula to a stirred 1: 1 saturated aqueous potassium sodium tartrate: saturated aqueous NaHCO at 23 deg.C₃In mixture (1L). The biphasic mixture was stirred for 1h and then the layers were separated. The aqueous layer was extracted with diethyl ether (500 mL). The combined organic layers were dried (MgSO)₄) Filtered and concentrated (rotary evaporation only, product is potentially volatile). The product was distilled under vacuum (bp100-120 ℃ C., 15mmHg) to give 7.58g (75.8mmol, 66%) of the desired product 2-propyl-prop-2-en-1-ol as a clear oil:¹HNMR(300MHz，CDCl3)5.22(s，1H)，5.06(s，1H)，4.27(s，2H)，2.24(t，J＝7.5Hz，2H)，1.75-1.60(m，2H)，1.12(t，J＝6.9Hz，3H).

2-bromomethyl-pent-1-ene 21d (R)⁹N-propyl).

To Et of n-propyl allyl alcohol (7.58g, 75.8mmol, 1equiv) at 0 deg.C₂Pyridine (0.58mL) was added to the O (65mL) solution. PBr was then added via cannula over 15min ₃Et (4.28mL, 45.5mmol, 0.6equiv)₂O (20mL) solution. After addition, the reaction was stirred at 0 ℃ for 75min, then the cold reaction mixture was slowly added to a stirred ice-cold saturated aqueous NaHCO₃(500 mL). The resulting biphasic mixture was extracted with diethyl ether (250 mL). The organic extract was washed with saturated aqueous NaHCO₃(2X 100mL), brine (100mL), 1.0N HCl (100mL), brine (100mL), washed and dried (MgSO 4)₄) Filtered and concentrated (0 ℃ rotary evaporation, product is volatile). The product was purified by flash column chromatography on silica gel using pentane as eluent to give 5.97g (36.8mmol, 49%) of the desired product 21d (R)⁹N-propyl) as a clear oil:¹HNMR(300MHz，CDCl₃)5.16(s，1H)，4.95(s，1H)，3.97(s，2H)，2.19(t，J＝7.5Hz，2H)，1.56-1.43(m，2H)，0.93(t，J＝7.8Hz，3H).

n-allyl glycine ethyl ester.

At 0 deg.C, inAllylamine 21e (R)^9bEt of (50mL, 666mmol, 2equiv) ═ H, m-1₂O (167mL) solution was added ethyl bromoacetate (36.9mL, 333mmol, 1 equiv). A white precipitate and an exothermic reaction were observed immediately after the addition; the exotherm resulted in solvent boiling for approximately 2 min. After addition, the reaction was stirred for 2.5h, then the ice-water bath was removed and the reaction was stirred at 23 ℃ overnight. After 15h at 23 ℃, the reaction mixture was filtered through a frit to remove the precipitated allylamine hydrobromide by-product. The solid was collected and used for Et ₂O (200mL) wash, then combine filtrates and concentrate. The product was distilled under vacuum (bp 48-55 ℃, 1.0mmHg) to give 35.5g (249mmol, 75%) of the desired product N-allylglycine ethyl ester as a yellow oil:¹HNMR(300MHz，CDCl₃)5.93-5.79(m，1H)，5.22-5.08(m，2H)，4.18(q，J＝7.2Hz，2H)，3.39(s，2H)，3.29-3.23(m，2H)，1.27(t，J＝7.2Hz，3H)；MS(ESPOS)：144.1[M+H]+.

n-allyl Glycine Ethyl ester hydrochloride 21f (R)^9b＝H，m＝1).

To Et N-allylglycine ethyl ester (10.0g, 70.0mmol, 1equiv) at 23 ℃ over 35min via an addition funnel₂O (260mL) in hexane (1.3L) was slowly added 4.0M HCl in dioxane (16.6mL, 66.5mmol, 0.95 equiv). After addition, the suspension was stirred for an additional 40min, then the product was isolated by frit filtration, washing with hexane (200 mL). A white solid was collected, transferred to a flask, and placed under vacuum (0.5mmHg) for 1h to give 11.5g of the desired product as a white solid. The reaction was repeated on an equal scale to give a total of 22.73g (127mmol, 90%) of the desired amine hydrochloride 21f (R)^9bH, m ═ 1), as a white solid:

¹HNMR(300MHz，DMSO-d6)9.50(s，2H)，5.95-5.81(m，1H)，5.49-5.37(m，2H)，4.21(q，J＝7.2Hz，2H)，3.92(s，2H)，3.59(d，J＝6.6Hz，2H)，1.24(t，J＝7.2Hz，3H)；MS(ESPOS)：144.1[M+H]+.

pseudoephedrine N-allylglycinamide 21h (R)^9b＝H，m＝1).

To contain 21f (R)^9bTHF (130mL) was added to a flask of (20.3g, 113mmol, 1.3equiv) and (1R, 2R) -pseudoephedrine 21g (14.4g, 86.9mmol, 1 equiv). The resulting mixture was stirred vigorously at 20 ℃ for 20 minutes to give a homogeneous slurry, which was then treated with solid lithium tert-butoxide (9.74g, 122mmol, 1.4equiv) added in one portion. The reaction was stirred at 20 ℃ for 2 days, after which analysis revealed that both starting materials were still present. Incomplete reaction is carried out by H ₂O (200mL), then THF was removed in vacuo. Using CH for the obtained aqueous solution₂Cl₂Extraction (2X 150mL), followed by saturation with NaC l and further CH₂Cl₂Extraction (2X 100 mL). Drying the organic extract (K)₂CO₃) Filtering, and concentrating. The crude product was purified by flash column chromatography on silica gel using 2: 96MeOH/Et₃N/CH₂Cl₂As eluent, 18g of product was obtained. This product, still substantially contaminated with N-allylglycine ethyl ester, was removed by gentle heating (60 ℃) under vacuum (1.0mmHg) for 15h to give 14.88g (56.8mmol, 65%) of the desired glycinamide product 21h (R)^9bH, m ═ 1), viscous oils:¹H NMR(300MHz，CDCl₃) (spectrum shows rotamers) 7.41-7.24(m, 5H), 6.00-5.80(m, 1H), 5.29-5.07(m, 2H), 4.64-4.44(m, 1H), 3.96-3.84(m, 0.5H), 3.63(d, J ═ 13.8Hz, 0.5H), 3.45-3.21(m, 4H), 2.95(s, 1.5H), 2.78(s, 1.5H), 1.11(d, J ═ 6.9Hz, 1.5H), 0.98(d, J ═ 6.9Hz, 1.5H); ms (espos): 263.2[ M + H]+.

HPLC (Symmetry C18, 3.5 μm particle size,pore diameter of 4.6mm, diameter of 30mm and length of 2% -98% of MeCN in H₂W/0.1% TFA in O over 10min, 2mL/min flow): rt 3.10min

Alkylation of pseudoephedrine N-allylglycinamide

To a solution containing LiCl (vacuum flame dried, 3.14g, 74.1mmol, 4equiv) was added to the flask pseudoephedrine N-allylglycinamide 21h (R)^9bH, m ═ 1) (4.85g, 18.5mmol, 1equiv) in THF (50 mL). The resulting mixture was stirred at 0 ℃ for 25min, then treated with LiHMDS solution (1.0M THF solution, 37.0mL, 37mmol, 2equiv) added slowly via cannula over 40 min. After addition of LiHMDS, the enolate solution was stirred at 0 ℃ for an additional 30min, then allyl bromide (3.00g, 18.5mmol, 1equiv) was added dropwise via syringe over 30 sec. The reaction was stirred at 0 ℃ for another 90min, then with H₂O (200mL) quench with CH₂Cl₂Extraction (3X 150 mL). Drying the organic extract (K)₂CO₃) Filtered and concentrated to give 8.0g of a yellow oil. A small portion of the crude product was purified by flash column chromatography on silica gel using 3: 2: 95MeOH/Et₃N/CH₂Cl₂As eluent, an analytically pure product sample was obtained. The remaining product was used without any purification in the next step:¹H NMR(300MHz，CDCl₃) (spectrum shows rotamers) 7.40-7.24(m, 5H), 5.90-5.76(m, 1H), 5.20-5.02(m, 2H), 4.92-4.75(m, 2H), 4.66-4.45(m, 2H), 4.20-4.00(m, 1H), 3.62(t, J ═ 6.3Hz, 1H), 3.34-3.16(m, 1H), 3.05-2.94(m, 2H), 2.84(s, 3H), 2.55(q, J ═ 7.2Hz, 1H), 2.22-1.80(m, 5H), 1.58-1.36(m, 3H), 1.11(d, J ═ 6.9Hz, 2H), 1.04(t, J ═ 72Hz, 1H, 0.96 (J ═ 6.9H), 1.89 (t, 2H), 1.89 (t, 7H); ms (espos): 345.0[ M + H [ ] ]+.

HPLC (Symmetry C18, 3.5 μm particle size,pore diameter of 4.6mm, diameter of 30mm and length of 2% -98% of MeCN in H₂W/0.1% TFA in O over 10min, 2mL/min flow): rt 4.28min

Boc-protection (R) of diene aminoamides 21i⁹Is n-propyl, R^9b＝H，m＝1)

To the amine (crude product from previous step, 8.0g, ca. 18mmol, 1equiv) in CH at 23 deg.C₂Cl₂(100mL) solution Triethyl acetate was addedAmine (2.83mL, 20mmol, 1.1equiv), followed by (Boc)₂O (8.07g, 37mmol, 2 equiv). The resulting mixture was stirred at 23 ℃ for 13.5h, then concentrated. The crude product was purified by flash column chromatography on silica gel gradient (column packed 5.5cm diameter by 17cm height) eluting with 25% EtOAc in hexane (1L), then 30% EtOAc in hexane (600mL) and then 40% EtOAc in hexane (400 mL). To give 21i (R)⁹Is n-propyl, R^9bH, m ═ 1)5.20g (11.7mmol, 65%, 2 steps) of pure product. Some mixed fractions were discarded, which also contained trace amounts of product:

¹H NMR(300MHz，CDCl₃) (spectrum shows rotamers) 7.52-7.24(m, 5H), 5.90-5.62(m, 1H), 5.44(t, J ═ 6.9Hz, 0.5H), 5.20-4.96(m, 2.5H), 4.77(d, J ═ 13.2Hz, 2H), 4.68-4.35(m, 2H), 4.00-3.55(m, 1H), 3.79(d, J ═ 5.7Hz, 1H), 2.91(s, 1H), 2.87(s, 2H), 2.52-2.29(m, 2H), 2.10-1.96(m, 2H), 1.54-1.35(m, 9H), 1.13-1.00(m, 2H), 0.96-0.86(m, 3H); ms (espos): 467.3[ M + Na ] ]+.

HPLC (Symmetry C18, 3.5 μm particle size,pore diameter of 4.6mm, diameter of 30mm and length of 2% -98% of MeCN in H₂W/0.1% TFA in O over 10min, 2mL/min flow): rt 6.85min

Ring blocked diene metathesis 21j (R)⁹Is n-propyl, R^9b＝H，m＝1).

At 23 ℃ to a diene 21i (R)⁹Is n-propyl, R^9bH) (5.20g, 11.7mmol, 1equiv) CH₂Cl₂(700mL) solution was added to benzylidene [1, 3-bis (2, 4, 6-trimethylphenyl) -2-imidazolidinylidene]Dichloro- (tricyclohexylphosphine) ruthenium (Grubbs generation 2 catalyst, 320mg, 0.38mmol, 0.03 equiv). The reaction was refluxed for 2h, then cooled to 23 ℃ and concentrated. The resulting product was first purified by flash column chromatography on silica gel (40% EtOAc in hexanes as eluent) to afford the desired product, which was still slightly unidentifiedThe material of (2) is contaminated. The product was then dissolved in hot hexane (100mL) and crystallized over 2 days. The crystallized product was isolated by frit filtration and washed with ice-cold hexane (100mL) to give 3.425g of the desired tetrahydropyridine (8.23mmol, 70%). The mother liquor was concentrated to give 0.57g of a brown oil which was subjected again to flash column chromatography on silica gel (40-50% EtOAc in hexane as eluent) to give a further 392mg (0.94mmol, 8%) of the desired product 21j (R) ⁹Is n-propyl, R^9b＝H，m＝1)：

¹H NMR(300MHz，CDCl₃) (spectrum shows rotamers) 7.50-7.25(m, 5H), 5.52-5.26(m, 1H), 5.05-4.96(m, 1H), 4.634.35(m, 2H), 4.30-358(m, 3H), 2.91(s, 3H), 2.50-2.34(m, 1H), 2.20-1.94(m, 3H), 1.46(s, 7H), 1.41(s, 2H), 1.19-1.01(m, 2H), 0.94-0.85(m, 3H); ms (espos): 439.3[ M + Na]+.

HPLC (Symmetry C18, 3.5 μm particle size,pore diameter of 4.6mm, diameter of 30mm and length of 2% -98% of MeCN in H₂W/0.1% TFA in O over 10min, 2mL/min flow): rt 6.29min

Cleavage (R) of pseudoephedrine adjuvant 21k⁹Is n-propyl, R^9b＝H，m ＝1).

To amide 21j (R) at 23 deg.C⁹Is n-propyl, R^9bH, M ═ 1) (3.42g, 8.22mmol, 1equiv) in MeOH (170mL) was added 1.0M aqueous NaOH (41.1mL, 41.1mmol, 5 equiv). The reaction was refluxed for 24h (oil bath temperature 100 ℃) then cooled to 23 ℃ and concentrated via rotary evaporation to remove most of the MeOH. The resulting aqueous solution was transferred to a separatory funnel using H₂Diluted O (100mL) with Et₂O (100 mL). The ether extract was washed with 0.5M aqueous NaOH (70mL) and then discarded. The basic aqueous layers were combined, acidified to pH 2 with 1.0N HCl, and extracted with EtOAc (2X 200 mL). The organic extracts were dried (MgSO)₄) Filtered and concentrated to give 2.46g of the desired Boc-protected amino acid 2 1k(R⁹Is n-propyl, R^9b＝H，m＝1)：¹HNMR(300MHz，CDCl₃) (spectrum shows rotamers) 5.36(d, J ═ 22.8Hz, 1H), 5.09(d, J ═ 4.8Hz, 0.5H), 4.90(br s, 0.5H), 4.14-3.97(m, 1H), 3.83-3.67(m, 1H), 2.57-2.37(m, 2H), 1.98(t, J ═ 7.2Hz, 2H), 1.48(s, 6H), 1.47-1.35(m, 2H), 1.46(s, 3H), 0.86(t, J ═ 72Hz, 3H); ms (espos): 292.1[ M + Na ]]+；MS(ESNEG)：268.2[M-H]-.

Method T

SoCl was added dropwise to anhydrous MeOH (20mL) at 0 deg.C₂(1.58mL, 21.6mmol), the solution was stirred at 0 ℃ for 10 minutes, then solid L-2-amino-4-pentenoic acid 22a (R) was added^9bH) (Aldrich) (1.0g, 8.7 mmol). The reaction mixture was stirred at ambient temperature for 48h and the solvent was removed in vacuo. Purification by column chromatography on silica gel (10% MeOH/DCM) gave L-2-amino-4-pentenoic acid methyl ester 22b (R)^9b＝H)(0.95g，85％)。

To a solution of methyl L-2-amino-4-pentenoate in dichloroethane (32mL) was added 2, 4, 6-collidine (2.3mL, 19.1mmol, 2.2equiv) and solid 2-nitrobenzenesulfonyl chloride at 0 ℃. The reaction was stirred at r.t. for 3 h. The solvent was removed in vacuo and the residue was taken up in ethyl acetate (200mL) and saturated aqueous NH₄Partition between Cl. The organic layer was washed with 1.0M aq₄Saturated aq₃Washed with brine and dried (MgSO) ₄) And concentrated to give a residue, which was purified by silica column chromatography (gradient: 10 to 20% EtOAc/hexanes) to provide the desired product 22b (R)^9bH)0.70g (26%) as yellow oil.

¹HNMR(300MHz，CDCl₃)δ8.10-8.06(m，1)，7.95-7.92(m，1)，7.76-7.73(m，2)，6.08(d，J＝8.2，1)，5.74-5.60(m，1)，5.17-5.12(m，2)，4.33-4.26(m，1)，3.52(s，3)，2.58(dd，J＝6.0，6.0，1)，3.44-3.30(m，2)，2.25-2.10(m，2)，2.11(s，3)，2.00-1.88(m，1)，1.86-1.70(m，1)，1.44-1.25(m，6)，0.98-0.88(m，9H).

MS(ESNEG)：313.0[M-H]^-.

To the stirred sulfonamide 22b (R)^9b＝H)(685mg，2.18mmol)、Cs₂CO₃(710mg, 2.18mmol) and tetrabutylammonium bromide (702mg, 2.18mmol) in DMF (5.0mL) was added 3-methylenehex-1-yl-tosylate 22c (R)⁹Propyl) (702mg, 2.61 mmol; prepared as described in Kelvin h. yong et al, journal of Organic Chemistry, 2001, 66, 8248) in DMF (1.0mL) and the reaction mixture was heated to 60 ℃ overnight. The reaction solvent was removed by evaporation, the resulting residue was dissolved in EtOAc, washed with 10% aqueous citric acid and brine, and the organic phase was washed over MgSO₄Drying and concentration gave a residue which was purified by silica column chromatography (17% -20% EtOAc/hexanes) to give the desired product 22d (R)⁹Is propyl radical, R^9bH) (0.38g, 42%) as oil.

MS(ESPOS)：433[M+Na]⁺.

To 22d (R)⁹Is propyl radical, R^9bTo a solution of H) (0.38g, 0.92mmol) in anhydrous DCM (40mL) was added benzylidene [1, 3-bis (2, 4, 6-trimethylphenyl) -2-imidazolidinylidene]Dichloro- (Tricyclohexylphosphine) ruthenium (23.3mg, 0.0276mmol), and reaction mixture obtained in N₂Refluxing for 2.5hr, cooling to room temperature, and concentrating. The product was purified by flash column chromatography on silica gel (35% ethyl acetate/hexane) to afford the desired compound 22e (R) ⁹Is propyl radical, R^9b＝H)(0.29g，81％)。

MS(ESPOS)：383[M+Na]⁺.

To a stirred mixture of thiophenol (183. mu.L, 1.79mmol) and 7-methyl-1, 5, 7-triazabicyclo [4.4.0 ] via cannula]Dec-5-ene (214. mu.L, 1.49mmol) in anhydrous DMF (3mL) was added alkene 22e (R)⁹Is propyl radical, R^9bH) (228mg, 0.596mmol) in anhydrous DMF (3.0mL) and the resulting reaction mixture was taken up in N₂Stir for 1 hour then concentrate to a residue. The residue was dissolved in ether and stirred with 1N aqueous HCl (15.0mL) for 5 min. Washing the aqueous phase with ether and solid carbonic acidPotassium is adjusted to be alkaline. The resulting basic aqueous phase was extracted three times with ether. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, concentrated, cooled to 0 deg.C, treated with 2M HCl in ether (0.8mL), the resulting mixture was stirred for 5min, and evaporated to dryness to give the desired product 22f (R)⁹Is propyl radical, R^9bH) as the hydrochloride salt (144mg, 103%).

MS(ESPOS)：198[M+H]⁺.

To amine 22f (R)⁹Is propyl radical, R^9bH) (143mg, 0.61mmol) in anhydrous dichloromethane (2.0mL) triethylamine (170 μ L, 1.22mmol) and di-tert-butyl dicarbonate (350mg, 1.6mmol) were added. The resulting reaction mixture was allowed to stand at room temperature and N₂Stirring overnight, then evaporating to dryness and purifying by flash column chromatography on silica gel using 20% ethyl acetate in hexane as eluent to give 22g of the desired compound (R) ⁹Is propyl radical, R^9b＝H)(176mg，86％)。

MS(ESPOS)：320[M+Na]⁺.

22g of the ester (R)⁹Is propyl radical, R^9bH) (175mg, 0.59mmol) in dioxane/water (6: 1) (4mL) was added 1M aqueous lithium hydroxide (0.65mL, 0.648 mmol). The resulting reaction mixture was allowed to stand at room temperature and N₂The mixture was stirred overnight and the solvent was removed under reduced pressure. The residue was dissolved in water and washed with ether. The aqueous layer was acidified with 10% citric acid and extracted with ether. The organic layer was washed with brine, dried over sodium sulfate and evaporated to dryness to give the desired protected cyclic amino acid 22h (R)⁹Is propyl radical, R^9b＝H)(175mg，105％)。

MS(ESNEG)：292[M-H]^-.

General procedure U

To nitrone 23a (prepared as described in Dondoni Et al, synthetic communications, 1994, 24, 2537-2550) (5.96g, 16.4mmol, 1equiv) of Et at 23 deg.C₂Et was added to O (200mL) solution₂AlCl solution (1.0M heptane solution, 16.4mL, 16.4mmol, 1equiv). The reaction was stirred at 23 ℃ for 15min, then cooled to-78 ℃ and treated with cyclopropylmagnesium bromide solution (0.5M THF solution, 99mL, 49mmol, 3equiv) added via cannula over 25 min. After stirring at-78 ℃ for an additional 1.7h, the reaction was quenched with 1.0M aqueous NaOH (80mL) at low temperature. The resulting mixture was stirred at 23 ℃ for 25min, then transferred to a separatory funnel and the layers separated. The organic layer was washed with brine (100mL, using gentle stirring to avoid formation of an emulsion). The original aqueous layer was washed with Et ₂O extraction (3X 150mL) and washing of each extract with brine (100 mL). The combined organic layers were dried (MgSO)₄) Filtered and concentrated to give 5.89g (14.5mmol, 89%) of the desired product 23b (R)²⁰+R²¹Cyclopropane) as a white solid. The product was used without further purification.

MS(ESPOS)：406.0[M+H]

At 0 ℃ to hydroxylamine 23b (R)²⁰+R²¹Cyclopropane) (5.89g, 14.5mmol, 1equiv) with Et₃CH for N (12.2mL, 87.3mmol, 6equiv)₂Cl₂(200mL) solution methanesulfonyl chloride (2.25mL, 29.1mmol, 2equiv) was added. The reaction was stirred at 0 ℃ for 20min, then at 23 ℃ for an additional 25min, then added to 1.0M aqueous NaOH: brine (1: 1, 200 mL). The layers were separated and the aqueous layer was treated with CH₂Cl₂Extraction (2X 50mL) followed by combining the organic layers and drying (MgSO)₄) Filtering, and concentrating. The resulting residue was dissolved in 1: 1 EtOAc: hexane (200mL) with H₂O (150mL), saturated aqueous NaHCO₃(200mL), brine (150mL), and dried (MgSO)₄) Filtered and concentrated to give 5.95g of a brown oil, the major component of which is the desired imine.

MS(ESPOS)：388.2[M+H]

Gilard reagent T (2.84g, 16.9mmol, 1.1equiv) was added to a solution of crude imine (5.95g) in MeOH (150mL) at 23 ℃. After stirring for 70min, the solution was concentrated. The residue was taken up in EtOAc (150mL) with 1: 1H ₂O, brine and saturated aqueous NaHCO₃(150 mL). The layers were separated and the aqueous layer was washed with EtOAc (15)0mL) was extracted. The combined organic layers were dried (MgSO)₄) Filtered and concentrated to give 4.70g of a yellow oil, the major component of which is the desired amine.

MS(ESPOS)：300.0[M+H]⁺.

To the crude amine (4.70g) in CH with 2, 6-lutidine (7.31mL, 62.9mmol, 4equiv) at 0 deg.C₂Cl₂(200mL) solution trifluoroacetic anhydride (3.28mL, 23.6mmol, 1.5mmol) was added. The reaction was stirred at 0 ℃ for 1h, then at 23 ℃ for 3 h; then using H₂O (100mL) quench. The quenched reaction mixture was stirred for 10min, then partitioned between 1: 1 EtOAc: hexane (300mL) and brine (200 mL). The layers were separated and the aqueous layer was washed with 1.0N HCl (300mL), saturated aqueous NaHCO₃Washed (300mL), brine (200mL), and dried (MgSO)₄) Filtering, and concentrating. The product was purified by flash column chromatography on silica gel using 25% EtOAc in hexane as eluent to give 4.50g of the desired trifluoroacetamide 23c (R)²⁰+R²¹Cyclopropane) (11.3mmol, 78% from hydroxylamine).

MS(ESPOS)：418.0[M+Na]⁺

At 23 ℃ to diacetonide 23c (R)²⁰+R²¹Cyclopropane) (4.50g, 11.3mmol, 1equiv) was added aqueous TFA (80%, 100mL, pre-cooled to 0 ℃). The reaction was stirred at 23 ℃ for 35min and then concentrated to give 3.87g of a white solid whose main component was the desired deprotected galactose.

MS(ESPOS)：338.1[M+Na]⁺

Crude galactose (3.65g, 11.6mmol, 1equiv) was reacted with Et at 23 deg.C₃N (16.1mL, 116mmol, 10equiv) CH₂Cl₂(130mL) solution Ac was added₂O (7.65mL, 81.1mmol, 7equiv), followed by DMAP (141mg, 1.2mmol, 0.1 equiv). The reaction was stirred at 23 ℃ for 2h, then quenched with MeOH (5 mL). The quenched reaction mixture was stirred for 5min and then Et₂Dilution with O (300 mL). Subjecting the resulting solution to H₂O(2×300mL)1.0N HCl (300mL), saturated aqueous NaHCO₃Washed (300mL), brine (300mL), and dried (MgSO)₄) Filtered and concentrated to give 5.07g of acetylated product as a mixture of α/β and pyranose/furanose isomers.

MS(ESPOS)：506.1[M+Na]⁺.

To the CH of the peracetate isomer (5.07g) at 0 deg.C₂Cl₂A solution of HBr in acetic acid (33%, 30mL) was added (150 mL). The reaction was stirred at 0 ℃ for 30min and then warmed to 23 ℃. After stirring for a further 3.5h, the reaction mixture is taken up with CH₂Cl₂Diluted (50mL), with ice water (2X 300mL), ice cold 50% saturated aqueous NaHCO₃(2X 300mL), ice-cold 50% saturated brine (300mL), and dried (MgSO 4)₄) Filtered and concentrated to give 4.33g of α -bromide 23d (R)²⁰+R²¹Cyclopropane) (8.60mmol, 76% from diacetonide). The product was used without further purification.

To bromide 23d (R) at 23 deg.C²⁰+R²¹Cyclopropane) (4.33g, 8.60mmol, 1equiv) in AcOH (100mL) was added AgOAc (1.44g, 8.60mmol, 1 equiv). After stirring at 23 ℃ for 45min, the reaction mixture was washed with CH₂Cl₂(350mL) diluted with H₂O (2X 400mL), ice cold 50% saturated aqueous NaHCO₃(3X 300mL), brine (400mL), and dried (MgSO₄) Filtered and concentrated to give 3.76g (7.78mmol, 91%) of the desired β -acetate as a white foam. The product was used without further purification.

MS(ESPOS)：506.1[M+Na]

To beta-acetate (3.76g, 7.78mmol, 1equiv) in CH at 23 deg.C₂Cl₂(50mL) solution addition of PCl₅(1.70g, 8.17mmol, 1.05equiv), followed by BF₃·OEt₂(50. mu.L). After stirring for 1h, the reaction is run with CH₂Cl₂Diluted (300mL), with ice cold brine (500mL), ice cold 50% saturated aqueous NaHCO₃(2X 500mL), ice-cold brine (500mL)Dried (MgSO)₄) Filtered and concentrated to give 3.72g of the desired beta-chloride 23e (R)²⁰+R²¹Cyclopropane). The product was used without further purification.

MS(ESNEG)：458.2[M-H]

At 23 ℃ to galactosylchloride 23e (R)²⁰+R²¹Cyclopropane) (3.72g, 8.10mmol, 1equiv) in DMF (30mL) and HMPA (7.5mL) was added MeSNa (1.70g, 24.3mmol, 3 equiv). After stirring at 23 ℃ for 35min, the reaction mixture was taken up in Et ₂O (150mL) to 1: 1H₂Split between O/brine (70 mL). The layers were separated and the aqueous layer was washed with Et₂O (150mL) strip. The combined organic layers were dried (MgSO)₄) Filtering, and concentrating. The residue was dissolved in CH₂Cl₂(130mL) with Et₃N(11.3mL，81.0mmol，10equiv)、Ac₂O (5.35mL, 56.7mmol, 7equiv) and DMAP (99mg, 0.81mmol, 0.1 equiv). After stirring for 1h at 23 deg.C, the reaction was quenched with MeOH (3.0 mL). The quenched reaction mixture was stirred for 10min and then in Et₂O (200mL) and H₂Partition between O (200 mL). The layers were separated and the organic layer was washed with 1.0M aqueous HCl (200mL), saturated aqueous NaHCO₃Washed (200mL), brine (100mL), and dried (MgSO)₄) Filtering, and concentrating. The crude product was purified by flash column chromatography on silica gel using 30% EtOAc in hexane as eluent to give 2.03g (4.30mmol, 53%) of the desired product 23f (R)²⁰+R²¹As cyclopropane, R¹SMe) as a white solid.

¹H NMR(300MHz，CDCl₃)β6.52(br d，J＝9.3Hz，1H)，5.65(d，J＝5.4Hz，1H)，5.56(dd，J＝0.9，3.0Hz，1H)，5.27(dd，J＝5.4，11.1Hz，1H)，5.20(dd，J＝3.0，10.5Hz，1H)，4.43(dd，J＝0.9，7.5Hz，1H)，3.66(q，J＝9.0Hz，1H)，2.15(s，3H)，2.08(s，3H)，2.07(s，3H)，1.98(s，3H)，0.94-0.78(m，1H)，0.70-0.61(m，1H)，0.57-0.33(m，3H)；MS(ESPOS)：493.9[M+Na]；MS(ESNEG)：470.2[M-H]^-.

To triacetyl trifluoroacetamide 23f (R) at 23 deg.C²⁰+R²¹As cyclopropane, R¹SMe) (2.03g, 4.30mmol, 1equiv) in MeOH (35mL) was added 1.0M aqueous NaOH (43mL, 43mmol, 10 equiv). The reaction was stirred for 100min and then acidified to pH 2 with 1.0M aqueous HCl (48 mL). The resulting solution was concentrated to dryness in vacuo, then the residue was dissolved/suspended in EtOH (40mL) and filtered through a medium porosity frit to remove NaCl. The solid was washed with EtOH (2X 20 mL). The filtrates were combined and washed with Amberlite IRA400 (OH) ^-Type) resin (60mL MeOH containing resin bed) and resin (2X 20mL) transferred with MeOH. The resulting mixture was stirred at 23 ℃ for 1h and then filtered. Resin with MeOH (3X 100mL), CH₃CN (100mL) wash. The filtrates were combined and concentrated to give 1.04g of the desired galactoside 23g (R)²⁰+R²¹As cyclopropane, R¹SMe) as a white solid (4.19mmol, 97%). The product was used without further purification.

¹HNMR(300MHz，CD₃OD)β5.28(d，J＝5.7Hz，1H)，4.13-4.06(m，2H)，3.97(d，J＝6.6Hz，1H)，3.59(dd，J＝3.3，9.9Hz，1H)，2.33(dd，J＝6.9，9.0Hz，1H)，2.05(s，3H)，0.91-0.77(m，1H)，0.58-0.43(m，2H)，0.39-0.31(m，1H)，0.28-0.19(m，1H)；MS(ESPOS)：272.0[M+Na]；MS(ESNEG)：248.2[M-H].

General procedure V

Following the general procedure in scheme 24, to a solution of compound 1 hydrochloride salt (9.90mmol, 1equiv) in THF (70mL) at 23 deg.C was added H₂O (70mL) followed by KHCO₃(12.9mmol, 1.3equiv) followed by (Boc)₂O (12.9mmol, 1.3 equiv). After stirring for 5h, the reaction mixture was partitioned between brine (200mL) and EtOAc (300 mL). The organic layer was separated, washed with brine (150mL), and dried (MgSO)₄). The solvent is removed under vacuum and the crude product is usedPurification by column chromatography system (40+ M column, 40mm ID. times.150 mm) using a linear ladderDegree (75% EtOAc/hexanes-100% EtOAc), total 1.2L eluent, flow rate 50mL/min, gave carbamate 24a (8.91mmol, 90%).

To a solution of carbamate 24a (15.9mmol, 1equiv) in benzene (300mL) at 23 ℃ was added p-anisaldehyde dimethyl acetal (4.06mL, 23.8mmol, 1.5equiv), followed by PPTS (199mg, 0.79mmol, 0.05 equiv). The reaction mixture was heated to reflux. After 4h, a second portion of p-anisaldehyde dimethyl acetal (2.0mL, 11.7mmol, 0.74equiv) was added. After an additional 17h, a third portion of p-anisaldehyde dimethyl acetal (2.0mL, 11.7mmol, 0.74equiv) was added. After the last addition, the reaction was refluxed for an additional 3H, then cooled to 23 ℃ in EtOAc (300mL) with H ₂Partition between O (300 mL). The organic layer was washed with 50% saturated aqueous NaHCO₃Washed (300mL), brine (150mL), and dried (MgSO)₄) Filtering, and concentrating. The crude product was purified via flash column chromatography on silica eluting with 40% EtOAc in hexanes to give the acetal 24b (11.3mmol, 71%).

To a solution of alcohol 24b (4.82mmol, 1equiv) in trimethyl phosphate (60mL) at 0 deg.C was added pyridine (3.90mL, 48.2mmol, 10equiv), followed by POCl over 60sec₃(0.88mL, 9.65mmol, 2 equiv). Other acylating agents may be used in this step, such as acid anhydrides (R)¹¹CO)₂O or acyl chloride R¹¹COCl and in the presence of a suitable base to give different R¹¹An acyl substituent. After the addition, the reaction was maintained at 0 ℃ for 2h, and then triethylammonium dicarbonate buffer (1.0M, pH 8.5, 40mL) was carefully added to quench the reaction. Then H is added₂O (60mL), the resulting mixture was stirred at 0 ℃ for 30min, then warmed to 23 ℃. After stirring the quenched reaction mixture at 23 ℃ for 2h, the volatiles were removed in vacuo by gentle heating (40-45 ℃) in a water bath. The crude product was azeotropically dried by co-evaporation with DMF (3X 100mL) and then with toluene (150mL, bath temperature 40-45 ℃ C.) to give a white solid. Crude product 24c (R) ¹¹＝PO(OH)₂) Essentially contaminated with triethylammonium salt but used without further purification.

To the protected phosphate ester 24c (R) prepared as described above at 0 deg.C¹¹＝PO(OH)₂) (crude product from previous step, ca. 4.8mmol) in 1, 2-dichloroethane (600mL) was added H₂O (25mL) followed by TFA (200 mL). After the addition, the reaction was maintained at 0 ℃ for 5min and then warmed to 23 ℃. After stirring for 25min at 23 ℃, the volatiles were removed in vacuo to give 16.2g of an oil. The crude product was dissolved in 1: 1H₂O/MeOH (70mL), filtration, and preparative HPLC purification of the resulting solution (Waters)HR C18, 6 μm particle size,pore size, 40mm ID X200 mm, H containing 5-60% acetonitrile₂O w/0.1% AcOH over 30min at a flow rate of 75mL/min) to give the desired 2-phosphate 5 (R)¹¹＝PO(OH)₂) (3.10mmol, 64% based on free alcohol) as a white solid.

Method W

To a solution of β -lactam 25a (2.92g, 12.8mmol, 1 equiv; prepared from benzyl (S) - (-) -4-oxo-2-azetidine-carboxylate (Aldrich), as described by Baldwin et al, Tetrahedron 1990, 46, 4733) in THF (30mL) was added a LDA solution (2.0M, 14.0mL, 28.1mmol, 2.2equiv) via a syringe pump over 20min at 0 ℃. The reaction was stirred at 0 ℃ for 30 min. Crotyl bromide (85%, 2.89mL, 28.1mmol, 2.2equiv) was added dropwise over about 1.5min, and the mixture was stirred at 0 ℃ for 2h, then at 1.0M aqueous KHSO ₄Partition between (100mL) and EtOAc (100 mL). Separating the organic layer with 1.0M aqueous KHSO₄Washed (100mL), brine (100mL), and dried (MgSO)₄) Filtering, and concentrating to obtain 25b (R)⁹' -2-butenyl) 3.65g (100%) of a green-yellow solid. The product was used without further purification.

MS(ESNEG)：282.2[M-H]^-.

At 0 deg.CNext, trimethylsilyldiazomethane (2.0M Et)₂O solution, 25.0mL, 50mmol, 3.9equiv) was slowly added to the acid 25b (R⁹' -2-butenyl) (3.65g, 12.9mmol, 1equiv) in methanol (70 mL). Removal of the solvent under vacuum gave 3.53g (11.9mmol, 92%) of the desired ester product as a yellow oil. The product was used in the subsequent reaction without further purification.

At 23 ℃ to alkene 25c (R)⁹' -2-butenyl) (3.53g, 11.9mmol, 1equiv) in EtOAc (40mL) Pd/C (10 wt.%, 482mg) was added. The reaction vessel was charged with hydrogen (balloon) and the mixture was stirred vigorously. After 2.5h, the reaction mixture was filtered through a pad of celite. The celite was washed with EtOAc (200mL) and the filtrate was concentrated to give 3.51g (11.7mmol, 99%) of 25C (R)⁹═ butyl) as a yellow oil. The product was used without further purification.

MS(ESPOS)：300.4[M+H]⁺.

To N-TBS beta-lactam 25c (R) at 23 deg.C ⁹To a solution of (3.51g, 11.7mmol, 1equiv) in THF (50mL) was added Et₃N.3 HF (0.95mL, 5.85mmol, 0.5 equiv). After stirring at 23 ℃ for 60min, the reaction mixture was partitioned between 90% saturated brine (150mL) and EtOAc (200 mL). The organic layer was separated, washed with brine (150mL), and dried (MgSO)₄) Filtering, and concentrating. The product was purified by flash column chromatography on silica gel using 50% EtOAc in hexane as eluent to give 1.48g (8.0mmol, 68%) of 25d (R)⁹Butyl) as a clear oil.

MS(ESPOS)：578.3[3M+H]⁺.

At 23 ℃ via syringe to beta-lactam 25d (R) over 2min⁹To a solution of ═ butyl) (2.06g, 11.1mmol, 1equiv) in THF (150mL) was added LiAlH₄Solution (1.0m thf solution, 22.9mL, 22.9mmol, 2.06 equiv). After stirring at 0 ℃ for 10min, the reaction was warmed to 23 ℃ and stirred for 15min, then refluxed for 3 h. The mixture was then cooled to 0 ℃ and quenched as follows, with careful addition of H₂O (1.0mL) followed by 15% aqueousNaOH (1.0mL), then H₂O (2.5 mL). The resulting suspension was stirred at 23 ℃ for 1.5h, using Et₂Dilution with O (250mL), filtration through celite, and Et₂O (250mL) wash. The filtrate was concentrated to give 1.42g of the desired product 25e (R)⁹═ butyl) (9.93mmol, 89%) as clear oil. The product was used without further purification.

MS(ESPOS)：287.4[2M+H]⁺.

To amino alcohol 25e (R) at 23 deg.C⁹(1.41g, 9.86mmol, 1equiv) in dichloromethane (50mL) Boc was added₂O (2.59g, 11.9mmol, 1.2 equiv). After stirring at 23 ℃ for 2h, the reaction mixture was concentrated. The product was purified by flash column chromatography on silica gel using 33% EtOAc in hexane as eluent to give 1.53g (6.31mmol, 64%) of 25f (R)⁹Butyl) as a clear oil.

MS(ESPOS)：266.0[M+Na]⁺.

To NaIO4(8.81g, 41.2mmol, 10equiv) in H at 23 deg.C₂O (60mL) solution RuCl was added₃·xH₂O (350mg, catalytic amount), followed by alcohol 25f (R)⁹Butyl) (1.00g, 4.12mmol, 1equiv) in acetone (60 mL). The biphasic mixture was stirred at 23 ℃ for 30min, then extracted with EtOAc (250mL) and the organic layer was decanted. The aqueous residue was extracted with two more portions of EtOAc (2X 150 mL). The combined organic extracts were treated with 2-propanol (75mL) and stirred at 23 ℃. After stirring for 2h, the mixture was filtered through celite, washing with EtOAc (300 mL). The filtrate was concentrated to give 0.78g of the desired product 25g (R)⁹═ butyl) (3.04mmol, 74%) as dark oil. The product was used without further purification.

MS(ESPOS)：280.0[M+Na]⁺.

Method X

At 23 ℃ to alcohol 25f (R)⁹' -2-methyl-2-butenyl) (3.31g, 13.0mmol, 1equiv) in DMF (100mL) imidazole (2.21g, 32.5mmol, 2.5equiv) was added followed by TBSCl (2.93g, 19.5mmol, 1.5 equiv). The reaction was stirred for 35min, then quenched with MeOH (2.0 mL). After stirring for 5min, the resulting mixture was taken up in Et₂O (500mL) and H₂Partition between O (400 mL). Separating the organic layer with H₂O (400mL), brine (200mL), and dried (MgSO)₄) Filtering, and concentrating to obtain 26a (R)⁹' -2-methyl-2-butenyl) 4.13g (11.2mmol, 86%) of the desired product as a clear oil.

MS(ESPOS)：392.4[M+Na]⁺.

Intermediate 26a (R) was reacted at-78 deg.C⁹A solution of ═ 2-methyl-2-butenyl) (2.03g, 5.50mmol, 1equiv) in dichloromethane (80mL) was treated with ozone (1.2L/min) introduced through the gas dispersion tube until a blue color was observed (20 min). An oxygen stream (1.2L/min) was then passed through the reaction mixture to expel excess ozone. After 15min, the oxygen flow was stopped and PPh was added₃(2.16g, 8.25mmol, 1.5 equiv). The reaction mixture was stirred at-78 ℃ for 30min, then at 0 ℃ for 15min, then warmed to 23 ℃. After stirring at 23 ℃ for 10min, silica gel was added and the resulting mixture was concentrated to dryness under vacuum to give a free-flowing powder which was directly loaded onto a silica gel column. Flash column chromatography using 30-33% EtOAc in hexane as eluent afforded 1.52g (4.42mmol, 80%) of aldehyde 26b as a clear oil.

MS(ESPOS)：398.0[M+MeOH+Na]⁺.

To a suspension of cyclopropylmethyltriphenylphosphonium bromide (1.22g, 3.06mmol, 1.5equiv) in THF (10mL) was added NaHMDS solution (1.0M THF solution, 3.06mL, 3.06mmol, 1.5equiv) dropwise via syringe at 0 ℃ over 1 min. The resulting solution was stirred at 0 ℃ for 20min, then treated with a solution of aldehyde 26g (700mg, 2.04mmol, 1equiv) in THF (3.0 mL; 2X 1.0mL rinse) which was transferred via cannula. After 15min at 0 ℃, the reaction was warmed to 23 ℃ and stirred for another 10min, then saturated NH was used₄Cl (30mL) was quenched. The resulting mixture was taken up in Et₂O (120mL) and H₂Partition between O (50 mL). The organic layer was separated, washed with brine (50mL), and dried (MgSO)₄) Filtering and concentratingAnd (4) shrinking. Flash column chromatography using 10% EtOAc in hexane as eluent gave 588mg (1.54mmol, 76%) of 26c (R)⁹' -3-cyclopropyl-prop-3-enyl) as a clear oil.

MS(ESPOS)：404.3[M+Na]⁺.

At 23 ℃ to alkene 26c (R)⁹' -3-cyclopropyl-prop-3-enyl) (191mg, 0.50mmol, 1equiv) in dioxane (5.0mL) was added dipotassium azodicarboxylate (973mg, 5.01mmol, 10equiv), followed by slow addition of AcOH (573 μ L, 10.0mmol, 20equiv) in dioxane (5.0mL) via a syringe pump over 16 h. After the addition was complete, the reaction was stirred for another 6h, then via Et ₂O (150mL) was filtered through a frit to remove the precipitate. The resulting solution was taken up with saturated aqueous NaHCO₃(2X 100mL), brine (80mL), and dried (MgSO)₄) Filtering, and concentrating. The above procedure was repeated three times on the crude product to complete the conversion of the alkene to give 183mg (0.48mmol, 96%) of saturated product 26d (R)⁹3-cyclopropyl-propyl) as a clear oil.

MS(ESPOS)：406.0[M+Na]⁺.

To TBS ether 26d (R) at 23 deg.C⁹' -3-cyclopropyl-propyl) (190mg, 0.50mmol, 1equiv) in THF (10mL) was added TBAF solution (1.0M THF solution, 0.55mL, 0.55mmol, 1.1 equiv). The resulting solution was stirred at 23 ℃ for 40min and then in Et₂O (50mL) and H₂Partition between O (50 mL). The organic layer was separated, washed with brine (50mL), and dried (MgSO)₄) Filtered and concentrated to give 133mg (0.50mmol, 100%) of 26e (R)⁹3-cyclopropyl-propyl) as a clear oil.

MS(ESPOS)：290.2[M+Na]⁺.

26e to the desired carboxylic acid product 26f (R)⁹3-cyclopropyl-propyl) was carried out as described in the previous examples.

MS(NEG)：282[M-H]^-.

Method Y

Racemic 27a (R)⁹N-propyl) synthesis

To 10b (R)⁹N-propyl) (22g, 0.13mol) in methanol (30mL) and concentrated HCl (10mL) was added platinum (IV) oxide (5 g). The reaction mixture was hydrogenated at 50psi for 16h, over celite The catalyst was removed by filtration and the filtrate was evaporated to dryness. The crude piperidinecarboxylic acid was used without further purification.

19g of the crude piperidinecarboxylic acid residue were dissolved in acetonitrile (200mL), and tetramethylammoniumhydroxide 5H was added₂O (33g), the reaction mixture was stirred for 30 minutes, di-tert-butyl dicarbonate (39g, 0.46mol) was added, and the reaction mixture was stirred at room temperature for 72 hours. Adding additional tetramethylammonium hydroxide 5H₂O (8g) and di-tert-butyl dicarbonate (9g), and the reaction mixture is stirred for 24 h. The reaction solvent was removed in vacuo and the resulting oil was diluted with water (200mL) and washed with ether (200 mL). The aqueous portion was acidified with solid citric acid to pH 3-4 and then extracted with ethyl acetate (3X 200 mL). The organic layers were combined and MgSO₄Drying, filtering and removing the solvent to obtain 27a (R)⁹N-propyl) (19g, 77%) as yellow oil, crystallized upon standing.

¹H NMR(300MHz，CD₃OD)δ4.31(m，1)，3.60(m，1)，3.33(m，1)，2.01(m，4)，1.24(m，14)，0.89(t，J＝5.7，3)；MS(ESNEG)：270[M-1]^-.

Subjecting racemic mixture 27a (R)⁹N-propyl) was diluted with acetonitrile (5 vol) and S- α -methylbenzylamine (0.5eq), and the mixture was heated to reflux, cooled, and seeded. The mixture was left overnight. The first batch of the salt formed is then filtered off until finally recrystallized.

The mother liquor from filtration was concentrated in vacuo and then dissolved in DCM (4 volumes) which was then washed with 1M citric acid (2/3 volumes of DCM). DCM was then separated, dried over magnesium sulfate, filtered and concentrated in vacuo. The 2R, 4S enantiomerically enriched free acid was dissolved in 5 volumes of acetonitrile, 0.85eq.r- α -methylbenzylamine was added, the mixture was heated to reflux, cooled and simultaneously seeded. The mixture was left overnight. The resulting salt is then filtered off for use (the ee of the salt is typically 85-90%).

The procedure for obtaining the second 2S, 4R salt is identical to the previous paragraph except that S- α -methylbenzylamine is used instead of R- α -methylbenzylamine. The resulting second salt (typically having an ee of about 80-90%) is then filtered off until finally recrystallized.

The mother liquor from filtration was concentrated in vacuo (2S, 4R salt to mother liquor, batch 2). The salt is decomposed (Broken) by the method described previously. At this stage, the impurities have been concentrated to such a level that salt formation is not possible. The free acid was subjected to column chromatography (30% EtOAc/hexanes) which removed the undesirable impurities. The column used a 10: 1 silica to compound weight ratio. The compound was also absorbed onto 3 equivalents (wt: wt) of silica.

The free acid from the column (enriched in the 2R, 4S enantiomer) was diluted with acetonitrile (5 vol), R- α -methylbenzylamine was added and recrystallization repeated.

Salt decomposition and 2S, 4R salt formation are equivalent to those described above. The salt formed in the third batch generally has an ee of 80-90%.

Three batches of 2S, 4R salt were combined and diluted with acetonitrile (7 vol). The mixture was heated to reflux at which time all the salt dissolved. The mixture was then allowed to cool to rt and left overnight with seeding. The salt that had precipitated out of solution was filtered. The salt showed about 97% ee. This procedure was repeated to give a salt with an ee of greater than 99%. The salt was dissolved in DCM (4 volumes), washed twice with 1M citric acid (ca 2/3 volumes of DCM), dried over magnesium sulfate, filtered and concentrated in vacuo. This procedure gave 77% of the theoretical yield of the 2S, 4R enantiomer as an amber oil in 98% ee.

Method Z

The following conditions are representative of the general coupling and deprotection scheme described in Process Z, where P¹＝H，P²Tert-butyl-carboxylate (Boc).

To azetidinoic acid 25f (R) at 23 deg.C⁹Butyl) (52mg, 0.20mmol, 1equiv), 7-Cl MTL 6b (R)²＝H，R³= Cl) (58mg, 0.20mmol, 1equiv) and HBTU (84mg, 0.22mmol, 1.1equiv) in DMF (2.0mL) was added DIPEA (88 μ L, 0.51mmol, 2.5 equiv). After stirring at 23 ℃ for 12h, the DMF was removed in vacuo and the residue was partitioned between EtOAc (100mL) and 1: 1 brine: 10% aqueous citric acid (100 mL). The organic layer was separated and washed with 1: 1 brine/saturated aqueous NaHCO₃Washed (100mL), brine (50mL), and dried (MgSO)₄) Filtered and concentrated to yield 82mg (0.17mmol, 84%) of 13a (R)²＝H，R³＝Cl，R⁹Is butyl, P¹＝H，P²T-butyl-carboxylate, m ═ 0), as a glassy solid, and was used in the next step without purification.

At 23 ℃ to carbamate 13a (R)²＝H，R³＝Cl，R⁹Is butyl, P¹＝H，P²Tert-butyl-carboxylate, m-0) (82mg, 0.17mmol, 1equiv) in 1, 2-dichloroethane (10mL) was added H₂O (0.40mL), followed by TFA (4.0 mL). After stirring at 23 ℃ for 20min, toluene (50mL) was added and the resulting solution was concentrated to dryness. The residue was purified by semi-preparative HPLC (Waters) HR C18, 6 μm particle size,pore size, 20mm ID X100 mm, H containing 5-60% acetonitrile₂O w/0.1% HCl, flow rate 20mL/min over 30 min) to give 41mg of the title compound 3-butyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -2-carboxylic acid]An amide ofA white solid.¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝6.0Hz，1H)，4.64(d，J＝7.8Hz，1H)，4.63-4.52(m，2H)，4.29(d，J＝10.2Hz，1H)，4.07(dd，J＝5.7，10.2Hz，1H)，4.00(t，J＝6.6Hz，1H)，3.82(d，J＝3.3Hz，1H)，3.75(dd，J＝8.4，9.9Hz，1H)，3.56(dd，J＝3.3，10.2Hz，1H)，2.92-2.76(m，1H)，2.14(s，3H)，1.90-1.67(m，2H)，1.45(d，J＝6.6Hz，3H)，1.44-1.24(m，4H)，0.93(t，J＝6.9Hz，3H)；MS(ESPOS)：411.0[M+H]⁺.

Method AA

To a solution of pyridine-2-carboxylic acid 10b (0.5mmol) in DMF (2mL) was added lincosamine (0.5mmol) as defined in general coupling scheme 13, followed by HBTU (214mg, 0.55mmol) and DIEA (132mg, 1 mmol). The reaction mixture was stirred at room temperature for 2 hr. The solvent was removed and the crude product was purified by silica gel column chromatography to give compound 13 b.

To a solution of pyridine 13b (0.46mmol) in water (10mL), AcOH (3mL) and MeOH (2mL) was added PtO₂(200mg), the resulting reaction mixture was shaken under 55psi of hydrogen overnight, or at a lower hydrogen pressure for a longer period of time. The residual catalyst was removed by celite filtration and the solvent was removed to give the crude product. Purification by column chromatography on silica eluting with MeOH in DCM afforded the lincosamide analog form 1 as defined in scheme 13.

In general, silica chromatography readily separates the cis-2S diastereomer from the undesired isomer. In some cases, separation of isomers requires semi-preparative HPLC. A representative set of conditions is as follows: waters HR C18 column, 6 μm particle size,pore size, 20mm ID X100 mm, 5-60% acetonitrile 0.1% AcOH/H₂O0.1% AcOH over 30min at a flow rate of 20 mL/min.

Method AB

[ 2-methyl-1- (3, 4, 5-tris-benzyloxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]-carbamic acid tert-butyl ester 15b (P)₁＝Boc，P₂＝Bn，R²＝Me，R³H) synthesis, step a (intermediate not shown)

To rapidly stirred 2a (P ═ Boc, R)¹＝Me，R²H) (2g, 5.7mmol) in benzene (40mL) was added 50% aqueous KOH (12.8mL), tetrabutylammonium hydrogen sulfate (0.67g), and benzyl bromide (6.77mL, 57.0mmol) in N₂Suspended under an atmosphere while vigorously stirring. After 3.5H, benzylamine (6.0mL) was added, the reaction mixture stirred an additional 20', then toluene (300mL) was added, and the organic layer was washed with H₂O(2×100mL)、2M KHSO₄(3×100mL)、sat.aq.NaHCO₃(1X 100mL), brine (1X 100mL), over MgSO₄Drying and evaporating to dryness. The crude product was chromatographed on silica, eluting with 10% EtOAc/hexanes to 15% EtOAc/hexanes to give the product 15b [ 2-methyl-1- (3, 4, 5-tris-benzyloxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl-step a]-tert-butyl carbamate as colorless foam (2.6g, 72%); ms (espos): 522.8[ M + H-Boc]⁺.

[ 2-methyl-1- (3, 4, 5-tris-benzyloxy-6-fluoro-tetrahydro-pyran-2-yl) -propyl ]-carbamic acid tert-butyl ester 15b (P)₁＝Boc，P₂＝Bn，R²＝Me，R³H) synthesis, step b)

To a stirred intermediate [ 2-methyl-1- (3, 4, 5-tris-benzyloxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -cooled to-16 deg.C]-tert-butyl carbamate (1.54g, 2.5mmol) in DCM (25mL) was added to DAST (0.599mL, 4.46mmol) in N₂Suspended under an atmosphere while vigorously stirring. After 5 min, solid NBS (0.599mL, 4.46mmol) was added over 5 min, the reaction mixture stirred an additional 45' and then DCM (300mL) was added and the organic layer was washed with sat₃(1X 100mL) and over MgSO₄Drying and evaporating to dryness. The crude product is subjected to a silica chromatography treatment,with 15% to 30% Et₂O/Hexane elution and isolation of the product as 1-alpha and beta fluorinated [ 2-methyl-1- (3, 4, 5-tri-benzyloxy-6-fluoro-tetrahydro-pyran-2-yl) -propyl]-mixture of tert-butyl carbamates 15b (P)₁＝Boc，P₂＝Bn，R²＝Me，R³H) as a colorless oil (0.85g, 58%); TLC (20% Et)₂O/Hexane) R_fIsomer 1 ═ 0.2, R_fIsomer 2 ═ 0.05.¹⁹F NMR(CDCl₃) Delta isomer 1: -132.78, -132.96, isomer 2: -145.05, -145.13, -145.23, -145.31.HPLC c183.5um, 4.6 × 30mm column, gradient eluent 2% -98% MeCN over 10 min; 1.5 mL/min: retention time 7.93min, 7.98min ms (ESPOS): 494.7[ M + H-Boc ]⁺；MS(ESNEG)：592.7[M-H]⁺.

2-methyl-1- (6-allyl-3, 4, 5-tri-benzyloxy-tetrahydro-pyran-2-yl) -propylamine 15c (P)₁＝H，P₂＝Bn，R¹═ allyl, R²＝Me，R³H) synthesis

In N₂To a stirred solution of intermediate 15b (831g, 2.5mmol) above in DCM (30mL) cooled to-32 deg.C under an atmosphere was added allyltrimethylsilane (1.12mL, 7.0 mmol). After 10 minutes, BF was added over 2 minutes₃·Et₂O (0.36mL, 2.8mmol), the reaction mixture was stirred for an additional 1.5h, then warmed to 0 ℃ for 30 minutes. Water (1mL) and TFA (15mL) were added to the reaction mixture, the reaction mixture was warmed to RT, stirred for 1h, and evaporated to dryness. Dissolve the residue in Et₂O (200mL) with 1M aq.K₂CO₃(50mL) and brine, over Na₂SO₄Drying, evaporating to dryness and isolating the product 15c (P)₁＝H，P₂＝Bn，R¹═ allyl, R²＝Me，R³H) as a colorless oil (0.69g, 96%); TLC (20% EtOAc/Hexane) R_f＝0.05；MS(ESPOS)：516.4[M+H-Boc]⁺.

2- (1-amino-2-methyl-propyl) -6-propyl-tetrahydro-pyran-3, 4, 5-triol

In N₂Next, 160mg of 2-methyl-1- (6-allyl-3, 4, 5-tris-benzyloxy-tetrahydro-pyran-2-yl) -propylamine 15c and 100mg of degusa 50% w/w wet 10% palladium on carbon were suspended in THF (5mL) and 1M aq. HCl (1mL), and the reaction mixture was brought to 1atm H₂Stirred under pressure for 24 h. The reaction mixture was filtered through celite and evaporated to dryness to give the product 2- (1-amino-2-methyl-propyl) -6-propyl-tetrahydro-pyran-3, 4, 5-triol (60.8mg, 89%) as the HCl salt. TLC (CHCl) ₃：MeOH：32％aq.AcOH)R_f＝0.35.MS(ESPOS)：248[M+H]⁺.

Method AC

In general, the final purification and isolation of the diastereomers of the compounds described in the following examples can be achieved by semi-preparative HPLC. The final product was passed through Waters PrepSystematic purification with WatersA dual λ absorbance detector set at 214nm, and s.e.d.e.r.e.A series of evaporative light scattering detectors. The general conditions for diastereomer separation are as follows: watersHR C18 column, 6 μm particle size,pore size, 20mm ID X100 mm, 5-60% acetonitrile 0.1% AcOH/H₂O0.1% AcOH over 30min at a flow rate of 20 mL/min. The collected fractions were pooled and lyophilized.

Examples

The following examples were prepared according to the above procedure.

Example 1

4- (3, 3-difluoro-allyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- (3, 3-difluoro-allyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]At 0 ℃ to 2b (R)²' ═ H) (45mg, 0.18mmol, 1equiv) in anhydrous DMF (0.5mL) was added triethylamine (79.4 μ L, 0.57mmol, 3.2equiv), followed by bis- (trimethylsilyl) trifluoroacetamide (71.2 μ L, 0.27mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. In a 25mL round bottom flask, the reaction mixture was added to the protected amino acid 8c (R) as prepared by general method M ⁹3, 3-difluoroallyl) (56mg, 0.19mmol, 1.1equiv), followed by the addition of solid HATU (91.2mg, 0.24mmol, 1.3 equiv). The reaction mixture was stirred at rt for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate (60mL), and washed with 10% citric acid (2X 40mL), water (40mL), half-saturated aq₃(40mL) and brine wash. Subjecting the organic layer to Na₂SO₄Drying and evaporating to obtain yellow syrup.

To a solution of the above crude coupling product in DCM (9mL) and methyl sulfide (0.2mL) was added trifluoroacetic acid (3mL) and water (0.2 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with toluene. The residue was purified by chromatography to give the title compound 4- (3, 3-difluoroallyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide (55.6mg, 73%) as a white solid:

¹H NMR(300MHz，CDCl₃)δ7.93(brs，1)，5.30(d，J＝4.8，1)，4.20-4.05(m，2)，3.96-3.77(m，3)，3.71-3.52(m，2)，3.19-3.07(m，1)，2.78-2.63(m，1)，2.38-2.21(m，1)，2.13(s，3)，2.20-1.97(m，4)，1.94-1.80(m，1)，0.92-0.84(m，6)；MS(ESPOS)：425.5[M+H]⁺；MS(ESNEG)：423.5[M-H]^-.

example 2

4- (3-pyridin-4-yl-allyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 2 the title compound 4- (3-pyridin-4-yl-allyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ]Amides were prepared according to the procedures described in example 1 and general procedure M, using ylides derived from triphenyl (4-pyridylmethyl) phosphonium chloride in the Wittig olefination step described in scheme 8. HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 10 min; 1.5 mL/min: rt is 2.99 min; ms (espos): 466.4[ M + H]⁺；MS(ESNEG)：464.2[M-H]^-，500.3[M+HCl]^-.

Example 3

4- (3-pyridin-4-yl-propyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 3 the title compound 4- (3-pyridin-4-yl-propyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]The amide is according toPrepared by general procedure M using a ylide derived from triphenyl (4-pyridylmethyl) phosphonium chloride in the Wittig olefination step described in scheme 8, followed by the protected amino acid 8b (R)⁹3-pyridin-4-yl-propyl, R²H) reductive deprotection. The coupling and deprotection process described in example 1 gives the desired end product. HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 10 min; 1.5 mL/min: rt is 2.99 min; ms (espos): 466.4[ M + H]⁺；MS(ESNEG)：468.3[M-H]^-，502.4[M+HCl]^-.

Example 4

4-butylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

N-Cbz- (2S), (4S) - (N-butylthio) proline 9d (P ═ Cbz, m ═ 1, R⁹Title intermediate was prepared as described in general procedure N. In N₂Next, to a stirred solution of intermediate 9b (P ═ Cbz, m ═ 1, LG ═ Ts) (1.34g, 3.08mmol) in DMF (10mL) was added 1-butanethiol (0.7mL, 6.16mmol, 2equiv), followed by 7-methyl-1, 5, 7-triazabicyclo [ 4.4.0: ═ b ═ m]Dec-5-ene (0.7mL, 4.87mmol, 1.6 equiv). After addition, the resulting mixture was stirred at room temperature for 18H, then in EtOAc and H₂And (4) distributing among the O. The organic layer was separated, washed with brine and then Na₂SO₄Dried, filtered and evaporated to dryness. The crude residue was chromatographed on silica, eluting with 3: 1 hexanes/EtOAc to give methyl ester 9c (P ═ Cbz, m ═ 1, R⁹N-butylthio) (470mg, 44%).

The methyl ester 9c was washed with lithium hydroxide (132.7mg, 3.16mmol, 2.4equiv) at 4: 1THF/H₂And treating in O overnight. The pH of the reaction solution was adjusted to 3 with aq.1M HCl and extracted with EtOAc (3X 100 mL). The organic phases are combinedOver Na₂SO₄Drying, filtration and evaporation to dryness gave the product 9d N-Cbz- (2S), (4S) - (n-butylthio) proline (463mg, quant.).

At 0 ℃ and N₂Next, to a solution of MTL 1a (140mg, 0.56mmol, 1equiv) in anhydrous DMF (2mL) was added triethylamine (0.3mL, 2.2mmol, 3.9equiv), followed by BSTFA (0.3mL, 1.1mmol, 2 equiv). The resulting mixture was stirred at 0 ℃ for 10min, then at room temperature for 30min, cooled to 0 ℃. A solution of the protected amino acid N-Cbz- (2S), (4S) - (N-butylthio) proline (215mg, 0.64mmol, 1.2equiv) in anhydrous DMF (1mL) was added, followed by solid HATU (320mg, 0.84mmol, 1.5equiv), the cooling bath was removed and stirred at room temperature for 2 h. After evaporation of the reaction mixture under high vacuum, the resulting residue was diluted with ethyl acetate (150mL), followed by 10% citric acid (2X 50mL), 0.5M sat₃(2X 50mL), brine (50mL), washed with Na₂SO₄Dried, filtered and evaporated to dryness. The obtained persilylated intermediate is used to contain50w-400x H⁺Resin (Aldrich) (200mg) in MeOH (60mL) for 45min, filtered, evaporated to dryness, and chromatographed on silica (92: 8 DCM/MeOH) to give the desired Cbz-protected lincosamide (185mg, 61%).

To a stirred suspension of 10% palladium on carbon (200mg) in anhydrous EtOH (6mL) under nitrogen was added 1, 4-cyclohexadiene (2mL) and after 10min a solution of the above Cbz-protected lincosamide (178mg, 0.33mmol) in EtOH (6mL) was added. The resulting mixture was stirred and heated to reflux overnight. After cooling, the reaction mixture was filtered through a pad of celite, washed with ethanol, the filtrate and washings were combined and evaporated to dryness. The resulting crude residue was chromatographed on silica (90: 9: 1 DCM/MeOH/concentrated ammonium hydroxide) to give the title compound, dissolved in 1: 1 acetonitrile/water (4mL), acidified (pH 4) with 1M HCl, and lyophilized to give the HCl salt (35mg) as a colorless powder: ms (espos): 439.3[ M + H ]⁺，461.2[M+Na]⁺.

Example 5

4-ethylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 5 the title compound is prepared according to general procedure N using sodium ethanethiol in the displacement step as depicted in scheme 9. The coupling and deprotection procedure described in example 4 gives the desired end product. Ms (espos): 412.6[ M + H]⁺.

Example 6

Example 6 the title compound was prepared according to general procedure N using triphenylphosphonium dibromide to install the 4- (S) bromide leaving group (P ═ Cbz, m ═ 1, LG ═ Br) in 9b, followed by the use of sodium ethanethiolate in the displacement step described in scheme 9. The coupling and deprotection procedure described in example 4 gives the desired end product. Ms (espos): 411.6[ M ]]⁺.

Example 7

4-ethylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 7 the title compound was prepared according to general procedure N using triphenylphosphonium dibromide to install the 4- (S) bromide leaving group (P ═ Cbz, m ═ 1, LG ═ Br) in 9b, followed by the use of sodium ethanethiolate in the displacement step described in scheme 9. The coupling and deprotection procedure described in example 4 gives the desired end product. Ms (espos): 429.1[ M + H ]⁺；MS(ESNEG)：427.6[M-H]^-，463.6[M+HCl]^-.

Example 8

Example 8 the title compound was prepared according to general procedure N, then sodium ethanethiol was used in the displacement step described in scheme 9. The coupling and deprotection procedure described in example 4 gives the desired end product. Ms (espos): 429.1[ M + H]⁺.

Example 9

4- (4-methyl-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

In N₂Next, to tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14₃CO，m＝1，R²＝H，R³OAc) (73.2mg, 92 μmol) in anhydrous DMF (300 μ L) was added 4-methylbenzyl mercaptan (Lancaster) (63.5 μ L, 0.46mmol) followed by MTBU (33.6 μ L, 0.23 mmol). The reaction mixture was stirred at rt for 16 h. The reaction mixture was dissolved in MeOH (1.5mL), 0.5M NaOMe in MeOH (920. mu.L, 9.2mmol) was added and the reaction mixture was stirred at room temperature for 18h, then added toResin bed (3.3mL in water). The resin was washed with methanol (5X 10mL), water (1X 10mL) and acetonitrile (2X 10mL), and the product was eluted with 5% concentrated NH₄OH in MeOH (5X 10mL) and MeCN (1X 10 mL). The combined washings were evaporated and subjected to preparative TLC treatment (95: 5 MeOH: 0.25M NH) ₃DCM) to give the title compound (25.0mg, 56%) as a colourless solid: ms (espos): 487.3[ M + H]⁺(ii) a HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 min: rt 1.91min.

Example 10

4- (4-fluoro-phenylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 10 the title compound was prepared by the procedure used in example 9 from tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14₃CO，m＝1，R²＝H，R³OAc). 4-Fluorothiophenol (Aldrich) is the nucleophile used in the displacement step: ms (espos): 477.3[ M + H]⁺.

Example 11

4- (3, 3, 3-trifluoro-propylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 11 the title compound was prepared by the procedure used in example 9 from tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14₃CO，m＝1，R²＝H，R³OAc). 1, 1, 1-Trifluoropropanethiol (Aldrich) is the nucleophile used in the displacement step. (300MHz, CDCl)₃)δ5.32(d，J＝5.8，1)，4.43(dd，J＝8.2，8.21)，4.36-4.31(m，1)，4.19-4.04(m，3)，3.90-3.g8(m，1)，3.78-3.55(m，3)，3.37-3.31(m，1)，2.96-2.82(m，3)，2.61-2.49(m，2)，2.12(s，3)，2.07-2.01(m，2)，1.14(d，J＝6.6，3)；(¹⁹F CDCl₃)δ-68.8t；MS(ESPOS)：479.2[M+H]⁺，963.3[2M+H]⁺.

Example 12

4- (3-methyl-butylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 12 the title compound was prepared by the procedure used in example 9 from tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14₃CO，m＝1，R²＝H，R³OAc). 3-methyl-butanethiol (Aldrich) is the nucleophile used in the displacement step: ms (espos): 453.3[ M + H]⁺(ii) a HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 10 min: rt 3.90min.

Example 13

4- (2, 4-dichloro-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 13 the title compound was prepared by the procedure used in example 9 from tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14₃CO，m＝1，R²＝H，R³OAc). 2, 4-dichlorobenzyl mercaptan (Maybridge) is the nucleophile used in the displacement step: ms (espos): 541.2[ M ]]⁺(ii) a HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 10 min: rt 4.383min.

Example 14

4- (thiophen-2-ylmethylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 14 the title compound was prepared by the procedure used in example 9 from tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14 ₃CO，m＝1，R²＝H，R³OAc). Thiophen-2-ylmethanethiol (Aldrich) is the nucleophile used in the displacement step: ms (espos): 479.2[ M + H]⁺(ii) a HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 10 min: rt is 3.656min.

Example 15

4- (pyrazin-2-ylmethylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 15 the title compound was prepared by the procedure used in example 9 from tosylate intermediate 14b (P ═ CF) prepared in general procedure R, scheme 14₃CO，m＝1，R²＝H，R³OAc). 2-Thiomethylpyrazine (Pyrazine Specialties Inc.) is the nucleophile used in the displacement step. Example 15 purification of the title compound by preparative TLC (16% ammonia in methanol/dichloromethane) gave the product 4- (pyrazin-2-ylmethylthio) -pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -2-carboxylate]Amide (9mg, 15%). Ms (espos): 475.5[ M + H]⁺；497.4[M+Na]⁺.

Example 16

4- (2, 4-dichloro-benzylsulfanyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

At 0 ℃ to 2b (R) ²' ═ H) (100mg, 0.40mmol, 1equiv) in anhydrous DMF (1mL) was added triethylamine (0.18mL, 1.27mmol, 3.2equiv), followed by addition of bis (trimethylsilyl) trifluoroacetamide (0.16mL, 0.60mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. The Boc-protected amino acid 9d prepared in general method N (P ═ Boc, m ═ 1, R) was added to the reaction mixture⁹2, 4-dichlorobenzylthio) (263mg, 0.65mmol, 1.63equiv), HATU (302mg, 0.80mmol, 2 equiv). The reaction mixture was stirred at rt for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate (150mL), and washed with 10% citric acid (2X 80mL), water (80mL), half-saturated NaHCO₃(80mL) and brine wash. Subjecting the organic layer to Na₂SO₄Drying and evaporating to obtain the required Boc-protected lincosamide,as a yellow syrup.

To a solution of the Boc-protected lincosamide in DCM (15mL) and methylsulfide (0.33mL) above was added trifluoroacetic acid (5mL) and water (0.33 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with toluene. The residue was purified by chromatography to give a white solid (61 mg). Purifying the white solid by preparative thin layer chromatography to obtain 4- (2, 4-dichloro-benzylthio) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylthio-tetrahydro-pyran-2-yl) -propyl ]Amide (47.5mg, 22%) as a white solid:¹HNMR(300MHz，CDCl₃)δ7.84(br s，1)，7.39-7.19(m，3)，5.31(d，J＝5.1，1)，4.09(dd，J＝5.4，9.9，1)，3.94-3.76(m，4)，3.81(s，2)，3.57-3.48(m，1)，3.40-3.32(m，1)，3.22-3.14(m，1)，2.88-2.79(m，1)，2.64-2.54(m，1)，2.33-2.22(m，1)，2.14(s，3)，1.93-1.85(m，1)，0.92-0.85(m，6).MS(ESPOS)：539.4[M+H]⁺.

example 17

4-butylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

9c(P＝Boc，m＝1，R⁹N-butyl sulfide) in N₂Next, to a solution of 9b (P ═ Boc, m ═ 1) (1.61g, 4.03mmol, 1equiv) in anhydrous DMF (12mL) prepared in general procedure N was added N-butylthiol (1.30mL, 12.1mmol, 3equiv), followed by the addition of 7-methyl-1, 5, 7-triazabicyclo [4.4.0 ″]Dec-5-ene (MTBE) (0.87mL, 6.05mmol, 1.5 equiv). The reaction mixture was stirred at rt overnight and concentrated to dryness. The residue was dissolved in ethyl acetate (100mL), washed with 10% citric acid (50mL) and brine, and concentrated. The residue was purified by chromatography to give a clear oil (1.24g, 97%):¹H NMR(300MHz，CDCl₃)δ4.30(t，J＝8.0，0.36)，4.23(t，J＝8.1，0.64)，4.00-3.94(m，0.64)，3.87-3.82(m，0.36)，3.72(s，1.1)，3.71(s，1.9)，3.29-3.15(m，2)，2.64-2.49(m，3)，1.97-1.84(m，1)，1.60-1.32(m，4)，1.44(s，3.2)，1.38(s，5.8)，0.93-0.86(m，3).

9c(P＝Boc，m＝1，R⁹to a solution of methyl ester 9c (1.24g, 3.91mmol, 1equiv) in THF (15mL) and water (5mL) was added lithium hydroxide monohydrate (0.82g, 19.55mmol, 5 equiv). The reaction mixture was stirred at rt overnight. THF was removed under vacuum. The residue was partitioned between ethyl acetate (200mL) and 10% citric acid (100 mL). The organic layer was washed with water (1X), brine (1X) and over Na₂SO₄Drying and evaporation gave clear oil 9c (P ═ Boc, m ═ 1, R) ⁹N-butyl) (1.21g, 100%): ms (espos): 204.4[ M-Boc + H]⁺，326.4[M+Na]⁺；MS(ESNEG)：302.3[M-H]^-.

4-butylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]At 0 ℃ to 2b (R)²' ═ H) (75mg, 0.30mmol, 1equiv) in anhydrous DMF (0.8mL) was added triethylamine (0.13mL, 0.96mmol, 3.2equiv), followed by the addition of bis- (trimethylsilyl) trifluoroacetamide (0.12mL, 0.45mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. To the reaction mixture was added Boc-protected amino acid 9c (P ═ Boc, m ═ 1, R⁹N-butylthio) (147mg, 0.49mmol, 1.63equiv), HATU (227mg, 0.60mmol, 2 equiv). The reaction mixture was stirred at rt for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate (100mL), and washed with 10% citric acid (2X 60mL), water (60mL), half-saturated NaHCO₃(60mL) and brine wash. Subjecting the organic layer to Na₂SO₄Drying and evaporating to obtain yellow syrup.

To a solution of the syrup in DCM (15mL) and methyl sulfide (0.33mL) was added trifluoroacetic acid (5mL) and water (0.33 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with toluene . The residue was purified by chromatography to give 4-butylsulfanyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl-ester as a white solid]-amide (95mg, 73%):¹HNMR(300MHz，CD₃OD)δ5.24(d，J＝6.0，1)，4.14-4.02(m，3)，3.94(dd，J＝7.1，8.9，1)，3.82(d，J＝3.3，1)，3.51(dd，J＝3.3，10.2，1)，3.45-3.32(m，2)，2.93(dd，J＝6.4，10.6，1)，2.71-2.55(m，3)，2.23-2.13(m，1)，2.10(s，3)，1.83-1.72(m，1)，1.63-1.52(m，2)，1.48-1.38(m，2)，0.97-0.88(m，9).MS(ESPOS)：437.5[M+H]⁺.

example 18

4-azido-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

N-Boc- (2S, 4R) -4-methanesulfonylproline methyl ester 9b (P ═ Boc, m ═ 1, LG ═ Ms.) to a solution of N-Boc- (2S, 4R) -4-hydroxyproline methyl ester (Bachem)9a (P ═ Boc, m ═ 1) (1g, 4.0mmol) in DCM (10mL) prepared in general method N, pyridine (1.64mL, 20.0mmol), methanesulfonyl chloride (0.631mL, 5.52mmol) were added, stirred at 0 ℃ for 2 hours, and further stirred at room temperature overnight. More DCM (100mL) was added, washed with HCl (1N, 50mL) and the organic portion was dried over magnesium sulfate. The solvent was removed to give the mesylate product (1.30g, 100%) which was used without further purification. N-Boc- (2S, 4S) -4-azidoproline methyl ester 9c (P ═ Boc, m ═ 1, R⁹N-Boc- (2S, 4R) -4-methanesulfonylproline methyl ester was dissolved in DMF (10mL), to which was added sodium azide (1.30g, 20.0mmol), and heated at 75-80 ℃ overnight. DMF was removed and the product was extracted with ethyl acetate (100mL) and washed with water (50 mL). Removal of the solvent gave the azide product 9c (P ═ Boc, m ═ 1, R) ⁹Azide) (0.98g, 90%).

9d(P＝Boc，m＝1，R⁹Stirring 9c (P ═ Boc, m ═ 1, R)⁹Azide) solution in THF (10mL) was treated with lithium hydroxide (300mg, 7.14mmol) in water (0.5mL) overnight. Excess solvent was removed by rotary evaporation and the residue was extracted with ethyl acetate and discarded. The aqueous portion was acidified, extracted with ethyl acetate and dried over magnesium sulfate. Removing the solvent to obtain the protected amino acid N-Boc- (2S, 4S) -4-azidoproline 9d (P ═ Boc, m ═ 1, R⁹Azide) (0.8g, 88%):

¹H NMR(300MHz，CD₃OD)δ4.34-4.24(m，2)，3.73-3.64(m，1)，3.39-3.34(m，1)，2.61-2.51(m，1)，2.16-2.09(m，1)，1.46(s，3)，1.42(s，6)；MS(ESNEG)：255(M-1).

1- (2- (S) -4- (S) - (azido) -N-pyrrolidin-2-yl- {1- (R) - [2- (S), 3- (S), 4- (S), 5- (R) -trihydroxy-6- (R) - (methylthio) tetrahydropyran-2-yl]-2-hydroxy-propan-1-yl } acetamide at 0 ℃ to 2b (R)⁹' ═ H) (200mg, 0.788mmol) in DMF (5mL) was added triethylamine (0.164mL, 1.18mmol) and bis- (trimethylsilyl) trifluoroacetamide (0.93mL, 3.94mmol), and the mixture was stirred at room temperature overnight. Then N-Boc- (2S, 4S) -4-azidoproline (300mg, 1.18mmol) and HATU (444mg, 1.18mmol) were added at 0 ℃ and stirred for 4 hours. At the end of the run, the DMF was removed and the residue was dissolved in ethyl acetate (100mL) and washed with citric acid (10%, 30mL), saturated sodium bicarbonate (30mL) and brine (30 mL). After drying the organic portion over sodium sulfate, the solvent was removed to give the crude product, which was used directly in the following deprotection step. To a solution of the crude product in dichloroethane were added 30% trifluoroacetic acid (10mL) and dimethylsulfide (0.5mL), and the reaction mixture was stirred for 1 hour. The solvent was removed and the crude product was chromatographed on silica gel eluting with 20% methanol in DCM to give the title compound as a white solid (278mg, 90%): TLC: rf ═ 0.38 (40% methanol in DCM);

TLC：R_f0.38 (40% methanol in DCM);¹H NMR(300MHz，CD₃OD)δ4.24(d，J＝5.4，1)，4.16(s，1)，4.03-4.13(m，3)，3.95(d，J＝3.6，1)，3.80(dd，J＝4.5，9.9，1)，3.51-3.56(dd，J＝3.3，10.2，1)，3.13-3.22(m，1)，2.95-3.00(m，1)，2.35-2.45(m，1)，2.08(s，3)，1.93-2.04(m，1)，1.29(t，J＝7.2，1)，0.97(m，3)；MS(ESPOS)：392[M+H]+.

example 19

4- [3- (furan-2-ylmethylsulfanyl) -propan-1-yl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- (3-hydroxy-propyl) -2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -piperidine-1-carboxylic acid tert-butyl ester.

At 0 ℃ to 2b (R)²' ═ H) (532mg, 1.85mmol, 1equiv) mixture in anhydrous DMF (4.5mL) triethylamine (1.28mL, 9.25mmol, 5equiv) was added, followed by the addition of bis- (trimethylsilyl) trifluoroacetamide (0.74mL, 2.78mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. To the reaction mixture was added the Boc-protected amino acid 11f (R) prepared in general procedure P⁹' -3-tert-butyldimethylsilyloxypropyl, P-Boc) (741mg, 1.85mmol, 1.0equiv) and HATU (886mg, 2.33mmol, 1.26 equiv). The reaction mixture was stirred at rt for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate and washed with 10% citric acid (1 ×), water (1 ×), sat₃(1x) and brine wash. Subjecting the organic layer to Na₂SO₄Drying and concentrating to obtain yellow syrup. The residue was dissolved in methanol (20mL) followed by addition Resin (340 mg).The mixture was stirred at rt for 1hr and filtered to remove the resin. The filtrate was concentrated and the residue was purified by column chromatography to give the product 4- (3-hydroxy-propyl) -2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl]-piperidine-1-carboxylic acid tert-butyl ester (694mg, 72%) as a white solid:

¹HNMR(300MHz，CDCl₃)δ5.33-5.28(m，1)，4.16-3.97(m，3)，3.89-3.69(m，3)，3.65-3.58(m，2)，3.56-3.47(m，1)，3.17-3.06(m，1)，2.33-2.23(m，1)，2.14(s，1.5)，2.13(s，1.5)，1.94-1.80(m，2)，1.67-1.50(m，5)，1.45(s，9)，1.42-1.23(m，2)，0.93-0.82(m，6).MS(ESPOS)：521.7[M+H]⁺.

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4- [3- (toluene-4-sulfonyloxy) -propyl ] -piperidine-1-carboxylic acid tert-butyl ester.

To a solution of 4- (3-hydroxy-propyl) -2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -piperidine-1-carboxylic acid tert-butyl ester (196mg, 0.38mmol, 1equiv) and p-toluenesulfonic anhydride (123mg, 0.38mmol, 1equiv) in DCM (1.5mL) was added triethylamine (63 μ L, 0.45mmol, 1.2equiv) dropwise at 0 ℃. The reaction mixture was stirred at 0 ℃ for 5hr, then diluted with ethyl acetate. The organic layer was washed with sat sodium bicarbonate, brine, dried and concentrated to give a white solid which was purified by chromatography to afford 11h (147.5mg, 58%) as a white solid:

¹H NMR(300MHz，CDCl₃)δ7.78-7.74(m，2)，7.35-7.31(m，2)，5.30(d，J＝5.7，1)，4.13-4.06(m，1)，4.03-3.93(m，3)，3.91-3.60(m，4)，3.54-3.45(m，1)，3.12-3.02(m，1)，2.43(s，3)，2.32-2.21(m，1)，2.121(s，1.7)，2.117(s，1.3)，1.83-1.73(m，2)，1.65-1.59(m，4)，1.45(s，5)，1.44(s，4)，1.37-1.15(m，3)，0.93-0.81(m，6)；MS(ESPOS)：675.9[M+H]⁺.

4- [3- (furan-2-ylmethylsulfanyl) -propyl ] -2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -piperidine-1-carboxylic acid tert-butyl ester.

In N₂To Boc-protected tosylate 2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl]-4- [3- (toluene-4-sulfonyloxy) -propyl](iv) -piperidine-1-carboxylic acid tert-butyl ester (91mg, 0.13mmol, 1equiv) in anhydrous DMF (0.42mL) was added furfuryl mercaptan (68. mu.L, 0.67mmol, 5equiv), followed by the addition of 7-methyl-1, 5, 7-triazabicyclo [4.4.0]Dec-5-ene (MTBE) (48. mu.L, 0.33mmol, 2.5 equiv). The reaction mixture was stirred at rt overnight, diluted with DCM, washed with brine (3 ×), dried, and concentrated. The residue was purified by preparative TLC (8% MeOH/DCM) to give the desired Boc-protected thioether 4- [3- (furan-2-ylmethylthio) -propyl]-2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl]-piperidine-1-carboxylic acid tert-butyl ester (63.3mg, 76%) as a clear syrup: ms (espos): 617.9[ M + H]⁺.

To Boc-protected thioether 4- [3- (furan-2-ylmethylthio) -propyl]-2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl]-piperidine-1-carboxylic acid tert-butyl ester in DCM (9mL) and methyl sulfide (0.2mL) was added trifluoroacetic acid (3mL) and water (0.2 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with toluene. The residue was purified by preparative TLC to give the title lincosamide product of example 19 (13mg, 25%) as a white solid: ¹HNMR(300MHz，CD₃OD)δ7.39(dd，J＝0.8，2.0，1)，6.32(dd，J＝2.1，3.3，1)，6.18(dd，J＝0.8，3.2，1)，5.24(d，J＝5.7，1)，4.17(dd，J＝3.2，10.1，1)，4.10-4.02(m，2)，3.79(d，J＝3.3，1)，3.71(s，2)，3.50(dd，J＝3.3，10.2，1)，3.43(dd，J＝2.9，11.9，1)，3.24-3.17(m，1)，2.78-2.67(m，1)，2.49(t，J＝7.1，2)，2.20-2.11(m，1)，2.10(s，3)，2.06-1.93(m，1)，1.78-1.70(m，1)，1.62-1.52(m，3)，1.38-1.29(m，2)，1.20-1.06(m，2)，0.91(d，J＝7.2，6)；MS(ESPOS)：517.8[M+H]⁺.

Example 20

4- (3-imidazol-1-yl-propan-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- (3-imidazol-1-yl-propan-1-yl) -2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -piperidine-1-carboxylic acid tert-butyl ester.

To a mixture of NaH (60%, 11.9mg, 0.30mmol, 2equiv) in anhydrous DMF (0.2mL) at 0 deg.C was added dropwise a solution of imidazole (40.4mg, 0.60mmol, 4equiv) in DMF (0.25 mL). The mixture was stirred at 0 ℃ for 10min and then cooled to-78 ℃. To the mixture was added dropwise a solution of Boc-protected tosylate (100mg, 0.15mmol, 1equiv) prepared in example 19 in anhydrous DMF (0.4 mL). The mixture was stirred at 0 ℃ for 2hr, then at rt overnight. The reaction mixture was diluted with DCM, washed with brine (3 ×), dried and concentrated. The residue was purified by chromatography to give the title Boc-protected imidazole compound (60mg, 71%) as a white solid. Ms (espos): 571.8[ M + H]⁺.

To a solution of the Boc-protected imidazole in DCM (9mL) and methylthio (0.2mL) was added trifluoroacetic acid (3mL) and water (0.2 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with toluene. The residue was purified by chromatography to give the title lincosamide compound of example 20 (10mg, 20%) as a white solid: ¹H NMR(300MHz，CD₃OD)δ7.63(s，1)，7.11(s，1)，6.95(s，1)，5.23(d，J＝5.7，1)，4.14(dd，J＝3.2，10.1，1)，4.10-3.99(m，4)，3.79(d，J＝3.6，1)，3.50(dd，J＝3.3，10.2，1)，3.27-3.21(m，1)，3.14-3.07(m，1)，2.64-2.54(m，1)，2.19-2.10(m，1)，2.10(s，3)，1.94-1.76(m，3)，1.70-1.64(m，1)，1.55-1.43(m，1)，1.30-1.18(m，2)，1.11-0.94(m，2)，0.92-0.88(m，6)；MS(ESPOS)：471.7[M+H]⁺.

Example 21

4- [3- (thiophen-2-ylsulfanyl) -propan-1-yl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- [3- (thiophen-2-ylsulfanyl) -propan-1-yl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

In N₂Next, to a solution of the protected tosylate (97mg, 0.14mmol, 1equiv) prepared in example 19 in anhydrous DMF (0.42mL) was added 2-thiophenethiol (Acros) (68. mu.L, 0.72mmol, 5equiv), followed by the addition of 7-methyl-1, 5, 7-triazabicyclo [4.4.0]Dec-5-ene (MTBE) (51.3. mu.L, 0.36mmol, 2.5 equiv). The reaction mixture was stirred at rt overnight, diluted with DCM, washed with brine (3 ×), dried, and concentrated. The residue was purified by preparative TLC (8% MeOH/DCM) to give a clear syrup (65.5mg, 74%): ms (espos): 619.8[ M + H]⁺.

To a solution of the syrup in DCM (9mL) and methyl sulfide (0.2mL) was added trifluoroacetic acid (3mL) and water (0.2 mL). The reaction mixture was stirred at rt for 1 h. The solvent was removed under vacuum and co-evaporated twice with toluene. The residue was purified by preparative TLC to give the title lincosamide compound of example 21 (16mg, 29%) as a white solid:

¹H NMR(300MHz，CD₃OD)δ7.45-7.42(m，1)，7.12-7.09(m，1)，7.01-6.96(m，1)，5.23(d，J＝5.7，1)，4.16(dd，J＝3.3，10.2，1)，4.10-4.01(m，2)，3.79(d，J＝3.3，1)，3.50(dd，J＝3.3，10.5，1)，3.34-3.28(m，1)，3.18-3.10(m，1)，2.76(t，J＝7.1，2)，2.68-2.58(m，1)，2.20-2.06(m，1)，2.10(s，3)，1.98-1.88(m，1)，1.71-1.28(m，6)，1.13-1.00(m，2)，0.90(d，J＝6.9，6)；MS(ESPOS)：519.7[M+H]⁺.

Example 22

4- (3-ethylsulfanyl-prop-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 22 the title compound was prepared by starting from the protected tosylate intermediate prepared in example 19, following the procedure used for examples 19-21, using sodium ethoxide in the displacement step: ms (espos): 465.3[ M + H]⁺.

Example 23

4- (3-cyano-prop-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 23 the title compound was prepared starting from the protected tosylate intermediate prepared in example 19, 11h, following the procedure used for examples 19-21, using a sodium cyanide nucleophile in the displacement step: ms (espos): 430.3[ M + H]⁺.

Example 24

4- (3-difluoromethylsulfanyl-propan-1-yl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

11c(R⁹' -3-hydroxy-propyne).

Propargyl alcohol (1.92mL, 33.0mmol, 2equiv) was added to a mixture of 4-iodopicolinic acid methyl ester 11b (4.36g, 16.5mmol, 1equiv), triphenylphosphine (346mg, 1.32mmol, 0.08equiv), cuprous iodide (251mg, 1.32mmol, 0.08equiv), palladium acetate (148mg, 0.66mmol, 0.04equiv) in triethylamine (60mL) at 23 deg.C, and the reaction mixture was stirred at 23 deg.C overnight. The reaction mixture was concentrated under high vacuum and the black residue was purified by column chromatography (2% MeOH in dichloromethane) to give a brown oil. The brown oil was again purified by column chromatography (100% EtOAc) to afford the desired product 11c (R) ⁹' -3-hydroxy-1-propyne) as a yellow oil (3.0g, 95%):

¹H NMR(300MHz，CDCl₃)8.70-8.74(dd，J＝0.9，5.1，1)，8.14(s，1)，7.46-7.50(dd，J＝1.8，5.1，1)，4.54(d，J＝6.3，2)，4.02(s，3)；MS(ESPOS)：192.1[M+H]；214.1[M+Na]；

HPLC: (Symmetry C183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 min; 1.5 mL/min): rt 1.42min.

11c(R⁹' -3-hydroxy-propyl).

At 23 ℃ to 11c (R)⁹' ═ 3-hydroxy-1-propyne) (2.0g, 10.5mmol, 1.0equiv) in MeOH (120mL) 20 wt.% Pd (OH) in water was added₂Carbon (1.0g), the reaction mixture was stirred under a hydrogen atmosphere overnight. The reaction mixture was filtered through celite, and the filtrate was concentrated to give the desired product 4- (3-hydroxy-propyl) -pyridine-2-carboxylic acid methyl ester 11C (R)⁹' -3-hydroxy-propyl) (2.03g, 99%) as a yellow oil:

¹H NMR(300MHz，CDCl₃)8.65(d，J＝5.1，1)，8.03(s，1)，7.34-7.36(dd，J＝1.8，5.1，1)，4.02(s，3)，3.71(t，J＝6.0，12.3，2)，2.83(t，J＝7.8，15.6，2)，1.92-1.97(m，2)；MS(ESPOS)：196.3[M+H]MS(ESNEG)：194.2[M-H]；

HPLC: (Symmetry C183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 min; 1.5 mL/min): rt 1.46min.

11d(R⁹' -3-hydroxy-propyl).

To 4- (3-hydroxy-propyl) -pyridine-2-carboxylic acid methyl ester 11c (R) at 23 deg.C⁹' ═ 1-hydroxypropyl) (1.81g, 9.28mmol, 1.0equiv) in MeOH (30mL) with H₂O (20mL) solution was added concentrated HCl (412. mu.L, 11.1mmol, 1.2equiv), followed by platinum (IV) oxide (600mg, 0.33 wt%), and the reaction mixture was stirred vigorously under 1atm hydrogen atmosphere for 48 h. The reaction mixture was filtered through celite, washing with methanol (200 mL). The filtrates were combined and concentrated under reduced pressure to give the desired product 11d (R) ⁹' -3-hydroxy-propyl) as HCl salt (2.02g, 8.52mmol, 91%): ms (espos): 202.2[ M + H]⁺.

11e(R⁹' -3-hydroxy-propyl, P ═ Cbz).

At 5 ℃ to 11d (R)⁹' -3-hydroxy-propyl) (2.02g, 8.52mmol, 1equiv) in dichloromethane (50mL) triethylamine (1.54mL, 11.07mmol, 1.3equiv) was added and the reaction mixture was stirred for 10 min. Benzyl chloroformate (1.55mL, 11.07mmol, 1.3equiv) was added to the solution, and the reaction mixture was stirred at 5 ℃ for 1h, then warmed to room temperature. The reaction mixture was concentrated and the crude product was partitioned between dichloromethane (250mL) and water (150 mL). Collecting the organic layer, passing through Na₂SO₄Drying and concentrating. The residue was purified by column chromatography (gradient from 50% to 75% EtOAc/hexanes) to afford the desired product 11e (R)⁹' -3-hydroxy-propyl, P ═ Cbz) (2.28g, 6.80mmol, 80%) as a yellow oil:

¹HNMR(300MHz，CDCl₃)7.34(s，5)，5.19(s，2)，4.48(t，J＝6.3，12.3，1)，3.68(bs，3)，3.60-3.64(m，2)，3.38-3.42(m，2)，1.94-1.98(m，3)，1.65-1.80(m，2)，1.33-1.37(m，2)，1.20-1.24(m，2)；MS(ESPOS)：358.0[M+Na]；

HPLC: (Symmetry C183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 min; 1.5 mL/min): rt 2.50min.

11e(R⁹' -3-methanesulfonylpropyl, P ═ Cbz).

To alcohol intermediate 11e (R) prepared by general procedure P at 0 deg.C⁹' -3-hydroxy-propyl, P ═ Cbz) (2.0g, 5.97mmol, 1equiv) in dichloromethane (15mL) triethylamine (1.0mL, 7.2mmol, 1.2equiv) was added and the reaction mixture was stirred for 15 min. Methanesulfonic anhydride (1.04g, 5.97mmol, 1equiv) was added to the solution and the reaction mixture was stirred at 0 ℃ for another 30 min. The reaction mixture was stirred in dichloromethane (250mL) with saturated aqueous NaHCO ₃(100 mL). Collecting the organic layer, passing through Na₂SO₄Drying and concentration gave the desired mesylate product 11e (R)⁹' -3-methanesulfonylpropyl, P ═ Cbz) (2.25g, 5.45 mmol; 91%) as clear oil:

¹H NMR(300MHz，CDCl₃)δ7.34(s，5)，5.19(s，2)，4.48(t，J＝6.3，12.3，1)，4.20(t，J＝6.3，14.1，2)，3.70(bs，3)，3.38-3.42(m，2)，3.02(s，3)，1.94-1.98(m，3)，1.65-1.80(m，2)，1.33-1.37(m，2)，1.20-1.24(m，2)；MS(ESPOS)：436.3[M+Na]；

HPLC：(c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 minutes; 1.5 mL/min): rt 2.86min.

11e(R⁹' -3-acetylthio-propyl, P ═ Cbz).

At 5 deg.C, to the nailSulfonate ester 11e (R)⁹' -3-methanesulfonylpropyl, P ═ Cbz) (2.25g, 5.45mmol, 1equiv) in DMF (30mL) was added potassium thioacetate (3.11g, 27.3mmol, 5equiv) and the reaction mixture was stirred at 5 ℃ overnight. The reaction mixture was washed with saturated aq.NaHCO in EtOAc (250mL)₃(100 mL). Collecting the organic layer, passing through Na₂SO₄Drying and concentrating. The resulting crude product was purified by column chromatography (25% EtOAc in hexane) to afford the desired thioester product 11e (R)⁹' -3-acetylthio-propyl, P ═ Cbz) (1.90g, 4.83mmol, 89%) as a yellow oil:

¹HNMR(300MHz，CDCl₃)δ7.34(s，5)，5.12(s，2)，4.46(t，J＝6.3，12.3，1)，3.69(bs，3)，3.38-3.42(m，2)，2.81-2.85(m，3)，2.32(s，3)，1.89-2.05(m，3)，1.65-1.80(m，2)，1.33-1.37(m，2)，1.20-1.24(m，2)；MS(ESPOS)：394.0[M+H]416.1[M+Na]；

HPLC：(c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 minutes; 1.5 mL/min): retention time 3.28min.

11f(R⁹' -3-difluoromethylthio-propyl, P ═ Cbz).

To 11e (R)⁹' -3-acetylthio-propyl, P ═ Cbz) (1.90g, 4.83mmol, 1equiv) in ethanol (8mL) 3N NaOH (4.5mL) was added. The reaction mixture was stirred at room temperature for 45min, then concentrated to give a clear oil. The resulting oil was dissolved in ethanol (20mL), the reaction mixture was deoxygenated via evacuation of the reaction flask, and then the reaction mixture was saturated with chlorodifluoromethane gas (Aldrich) at a pressure of 1 atm. The reaction mixture was stirred at 5 ℃ for 16h, then neutralized with 1N HCl at 0 ℃ and concentrated under reduced pressure. The residue was made alkaline with 0.5N aqueous NaOH and washed with ether. The aqueous layer was acidified to pH 2.0 with 1N HCl and extracted with ethyl acetate (3X 100 mL). The organic layer was washed with brine (2X 100mL) and dried (MgSO) ₄) And (4) concentrating. The resulting crude residue was purified by column chromatography (50% EtOAc/49% hexane/1% AcOH) to provide the desired difluoromethylthio product 11f (R)⁹' -3-difluoromethylthio-propyl, P ═ Cbz) (0.75g, 1.94mmol, 40%) as a clear oil:¹H NMR(300MHz，CDCl₃)δ7.36(s，5)，6.97(s)，6.79(s)，6.60(s)，5.15(s，2)，4.51(t，J＝6.3，12.3，1)，3.38-3.42(m，2)，2.75(t，J＝7.2，14.4，1)，2.47-2.51(m，1)，1.89-2.05(m，3)，1.65-1.80(m，2)，1.33-1.37(m，2)，1.21-1.23(m，2)；MS(ESPOS)：388.1[M+H]+410.1[M+Na]+；

HPLC：(c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 5 minutes; 1.5 mL/min): rt 2.85min.

11f(R⁹' -3-difluoromethylthio-propyl, P ═ Boc).

At 23 ℃ to 11f (R)⁹' -3-difluoromethylthio-propyl, P ═ Cbz) (750mg, 1.94mmol, 1equiv) in acetonitrile (100mL) was added iodotrimethylsilane (0.8mL, 5.81mmol, 3equiv) and the reaction mixture was stirred for 30 min. The reaction mixture was concentrated to give the crude deprotected product (491mg, 1.94mmol, 100%). To this were added dichloromethane (100mL), triethylamine (0.54mL, 3.88mmol, 2equiv), and di-tert-butyl dicarbonate (0.67mL, 2.91mmol, 1.5 equiv). The reaction mixture was stirred at 5 ℃ overnight. The reaction mixture was concentrated and the crude product was partitioned between dichloromethane (250mL) and water (150 mL). The organic layer was collected and dried (Na)₂SO₄) And (4) concentrating. The resulting crude residue was purified by column chromatography (50: 49: 1 EtOAc/hexane/AcOH) to provide the desired product 11f (R) ⁹' -3-difluoromethylthio-propyl, P-Boc) (671mg, 98%) as a clear oil:

¹H NMR(300MHz，CD₃OD)δ7.10(s)，6.90(s)，6.70(s)，4.19-4.23(m，1)，3.48-3.53(m，1)，2.68(t，J＝6.6，13.8，1)，2.56(t，J＝7.2，14.4，1)，1.80-1.95(m，4)，1.50-1.80(m，5)，1.33(s，9)，1.13-1.20(m，2)；MS(ESPOS)：254.1[M-Boc+H]MS(ESNEG)：352.2[M-H]-.

4- (3-difluoromethylsulfanyl-propyl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

To a stirred solution of 7-Cl-MTL 6b (R) at 23 deg.C²＝H，R³To a solution of 195mg (0.68 mmol, 1.2equiv) in DMF (2.5mL) was added diisopropylethylamine (0.3mL, 1.71mmol, 3equiv), followed by boc-protected amino acid 11f (R)⁹' -3-difluoromethylthio-propyl, P ═ Boc) (200mg, 0.57mmol, 1equiv) in DMF (2.5mL) and HBTU (324mg, 0.85mmol, 1.5 equiv). The resulting solution was stirred at room temperature for 3h, then concentrated to dryness. The solid residue was taken up in ethyl acetate (300mL) and saturated aq₃(100 mL). The organic layer was collected and dried (Na)₂SO₄) And (4) concentrating. To a portion of the residue (60mg, 0.12mmol, 1equiv) was added 1, 2-dichloroethane (5mL), water (0.2mL), followed by neat TFA (2.0mL) at 23 ℃, the reaction mixture was stirred at room temperature for 15min, then concentrated in vacuo. The crude product was purified by preparative HPLC to give the title compound of example 24 as a white powder:

¹H NMR(300MHz，CD₃OD)δ7.12(s)，6.93(s)，6.74(s)，5.21(d，J＝5.7，1)，4.48(d，J＝8.4，2)4.40(d，J＝10.2，1)，4.17(d，J＝9.6，1)，3.97-4.02(dd，J＝5.4，9.9，1)3.71(d，J＝3.3)，3.46-3.52(m，2)，2.71-2.76(t，J＝6.9，13.8，1)，2.05(s，3)，1.83(s，1)，1.61-1.77(m，5)，1.35(d，J＝6.9，4)，1.08-1.22(m，3)；MS(ESPOS)：507.1[M+H]⁺.

example 25

4- (3-difluoromethylthio-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- (3-difluoromethylsulfanyl-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

To stirred 2b (R) at 23 deg.C²' ═ H) in DFM (2.5mL) diisopropylethylamine (0.3mL, 1.7mmol, 3equiv) was added followed by Boc-protected amino acid 11f (R)⁹' -3-difluoromethylthio-propyl, P ═ Boc) (200mg, 0.57mmol, 1equiv) in DMF (2.5mL) and HBTU. The resulting solution was stirred at room temperature for 3h and concentrated to dryness. The solid residue was taken up in ethyl acetate (300mL) and saturated aq₃Are distributed among the devices. The organic layer was collected and dried (Na)₂SO₄) And (4) concentrating. To a portion of the residue (60mg, 0.12mmol, 1equiv) was added 1, 2-dichloroethane (5mL), water (0.2mL), followed by neat TFA (2.0mL) at 23 ℃, the reaction mixture was stirred at room temperature for 15min, then concentrated. The crude product was purified by preparative HPLC to afford the desired title compound of example 25 as a white powder:

¹HNMR(300MHz，CD₃OD)δ7.13(s)，6.94(s)，6.75(s)，5.16(d，J＝5.7，1)，4.12(d，J＝9.9，1)，3.97-4.02(dd，J＝4.8，10.2，2)，3.71(d，J＝3.3，1)，3.42-3.46(m，2)，2.88-2.96(m，2)，2.75(t，J＝7.8，15.0，2)，2.59-2.63(m，2)，2.03(s，3)，1.81-1.87(m，1)，1.61-1.70(m，5)，1.20-1.38(m，4)，0.82-0.84(d，J＝6.9，6)；MS(ESPOS)：487.1[M+H]⁺.

example 26

4- (2- [1, 3] dithiolan-2-yl-ethyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- (3, 3-diethoxy-prop-1-ynyl) -pyridine-2-carboxylic acid methyl ester.

To a dry flask was added intermediate 11b (2.98g, 11.33mmol, 1equiv) prepared in general procedure P, triphenylphosphine (238mg, 0.91mmol, 0.08equiv), copper (I) iodide (172.6mg, 0.91mmol, 0.08equiv), palladium acetate (101.6mg, 0.45mmol, 0.04equiv) and triethylamine (42 mL). The mixture was degassed with nitrogen, followed by the addition of 3, 3-diethoxy-propyne (Aldrich) (2.90g, 22.7mmol, 2 equiv). The mixture was stirred at rt for 3 h. The solvent was removed under vacuum to give a dark residue. The residue was chromatographed to give yellow oil 11c (R)⁹' -3, 3-diethoxy-prop-1-ynyl) (3g, 100%):

¹H NMR(300MHz，CDCl₃)δ8.69(dd，J＝0.8，5.0，1)，8.15(d，J＝0.8，1.4，1)，7.49(dd，J＝1.7，5.0，1)，5.48(s，1)，3.99(s，3)，3.82-3.73(m，2)，3.71-3.62(m，2)，1.26(t，J＝7.2，6).MS(ESPOS)：264.5[M+H]⁺.

4- (3, 3-diethoxy-1-propyl) -piperidine-1, 2-dicarboxylic acid 2-methyl ester 11d (R)⁹' -3, 3-diethoxy-1-propyl).

To 11c (R)⁹' -3, 3-diethoxy-prop-1-ynyl) (3g) in MeOH (15mL), acetic acid (15mL) and water (15mL) platinum oxide (1.0g) was added. The mixture was purged with hydrogen and aerated (50psi) and shaken at rt for 5 hr. The platinum oxide was removed by filtration and the filtrate was concentrated to give the desired product 11d (R)⁹' -3, 3-diethoxy-1-propyl) (2.45g, 79%) as oil: ms (espos): 296.5[ M + Na ]]⁺.

4- (3, 3-diethoxy-1-propyl) -piperidine-1, 2-dicarboxylic acid 1-allyl ester 2-methyl ester 11e (R) ⁹' -3, 3-diethoxy-1-propyl, P ═ Alloc).

At 0 ℃ to 11d (R)⁹' -3, 3-diethoxy-1-propyl) (2.4g, 8.79mmol, 1equiv) and pyridine (1.26mL, 11.9mmol, 1.35equiv) in THF (29mL) were added dropwise allyl chloroformate solution (0.96mL, 11.9mmol, 1.4 equiv). The mixture was slowly warmed to rt and stirred at rt for 3 hr. The solution was filtered and the solvent was removed under vacuum. The residue was purified by chromatography to give 11e (R)⁹' -3, 3-diethoxy-1-propyl, P ═ Alloc) (2.1g, 66%) as a clear oil: ms (espos): 380.6[ M + Na ]]⁺.

4- (3-oxo-propyl) -piperidine-1, 2-dicarboxylic acid 1-allyl ester 2-methyl ester.

Mixing 11e (R)⁹A solution of' -3, 3-diethoxy-1-propyl, P ═ Alloc) (2.03g) in acetic acid (32mL) and water (8mL) was stirred at rt overnight. The solvent was removed under high vacuum. The residue was diluted with ethyl acetate, washed with sat sodium bicarbonate (1 ×) and brine (1 ×). The organic layer was dried and concentrated. The residue was chromatographed to give methyl ester 11e (R)⁹' -3-oxo-propyl, P ═ Alloc) (1.2g, 75%) as a clear oil:

¹H NMR(300MHz，CDCl₃)δ9.75(t，J＝1.5，1)，5.96-5.82(m，1)，5.30-5.16(m，2)，4.57(d，J＝5.4，2)，4.46(t，J＝6.0，1)，3.74-3.65(m，1)，3.71(s，3)，3.42-3.32(m，1)，2.48-2.41(m，2)，2.02-1.35(m，7)；MS(ESPOS)：306.5[M+Na]⁺.

4- (2- [1, 3] dithiolan-2-yl-ethyl) -piperidine-1, 2-dicarboxylic acid 1-allyl ester 2-methyl ester.

Under nitrogen to 11e (R)⁹A mixture of 3-oxo-propyl, P Alloc (248mg, 0.87mmol, 1equiv) and 1, 2-ethanedithiol (0.147mL, 1.75mmol, 2equiv) was added with boron trifluoride-acetic acid complex (0.122mL, 0.87mmol, 1 equiv). The mixture was stirred vigorously for 1 hr. The mixture was diluted with hexane, washed with sat. sodium bicarbonate (3x) and brine (1 x). The organic layer was dried and concentrated. The residue was purified by chromatography to give 11e (R)⁹’＝2-[1，3]Dithiolan-2-yl-ethyl, P ═ Alloc) (144mg, 46%) as oil: ms (espos): 382.5[ M + Na ]]⁺.

4-(2-[1，3]Dithiolan-2-yl-ethyl) -piperidine-1, 2-dicarboxylic acid 1-allyl ester 11f (R)⁹’＝(2-[1，3]Dithiolan-2-yl-ethyl), P ═ Alloc).

To 11e (R)⁹’＝2-[1，3]Dithiolan-2-yl-ethyl, P ═ Alloc) (144mg, 0.40mmol, 1equiv) mixture in THF (3mL) and water (1mL) was added lithium hydroxide monohydrate (67mg, 1.6mmol, 4 equiv). The mixture was stirred at rt overnight. THF was removed under vacuum. The aqueous layer was dissolved in ethyl acetate and partitioned with 10% citric acid. The organic layer was washed with water (1x) and brine (1x), dried, and concentrated to give 11f (R)⁹’＝(2-[1，3]Dithiolan-2-yl-ethyl), P ═ Alloc) (127mg, 92%) as a syrup:

¹H NMR(300MHz，CDCl₃)δ5.96-5.83(m，1)，5.30-5.17(m，2)，4.59(d，J＝5.4，2)，4.48(t，J＝6.4，1)，4.41(t，J＝6.9，1)，3.75-3.64(m，1)，3.44-3.33(m，1)，3.25-3.12(m，4)，2.05-1.35(m，9).MS(ESPOS)：346.5[M+H]⁺.

4- (2- [1, 3] dithiolan-2-yl-ethyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

At 0 ℃ to 2b (R)²' ═ H) (95mg, 0.33mmol, 1equiv) mixture in anhydrous DMF (0.8mL) triethylamine (0.23mL, 1.65mmol, 5equiv) was added, followed by bis (trimethylsilyl) trifluoroacetamide (0.13mL, 0.49mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. Adding acid 11f (R) to the reaction mixture⁹’＝(2-[1，3]Dithiolan-2-yl-ethyl), P ═ Alloc) (115mg, 0.33mmol, 1.0equiv) and HATU (158mg, 0.42mmol, 1.3 equiv). The reaction mixture was stirred at rt for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate and washed with 10% citric acid (1 ×), water (1 ×), sat₃(1x) and brine wash. An organic layer is formedThrough Na₂SO₄Drying and concentrating. The residue was purified by chromatography to give Alloc-protected lincosamide analogue (133mg, 70%) as a syrup: ms (espos): 579.8[ M + H]⁺.

To a solution of Alloc-protected lincosamide described above (103mg, 0.18mmol, 1equiv) in THF (2.3mL) was added dimedone (0.25g, 1.78mmol, 10equiv) and tetrakis- (triphenylphosphine) palladium (41.1mg, 0.036mmol, 0.2 equiv). The mixture was stirred at rt overnight. The solvent was removed under vacuum and the residue was purified by chromatography to give the title compound of example 27 (34mg, 49%) as a yellowish solid:

¹H NMR(300MHz，CD₃OD)δ5.23(d，J＝5.7，1)，4.45(t，J＝6.9，1)，4.18-4.01(m，3)，3.81-3.77(m，1)，3.56-3.47(m，2)，3.25-3.07(m，5)，2.66-2.56(m，1)，2.18-2.13(m，1)，2.10(s，3)，1.94-1.63(m，4)，1.55-1.35(m，3)，1.12-0.98(m，2)，0.95-0.88(m，6)；MS(ESPOS)：495.6[M+H]⁺.

Example 27

Example 27 was not included.

Example 28

4- [2- (4-methyl-thiazol-2-yl) -ethyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4- [2- (4-methyl-thiazol-2-yl) -ethyl]1-allyl-piperidine-1, 2-dicarboxylate 11f (R)⁹' -4-methyl-thiazol-2-yl, P ═ Alloc).

This intermediate was prepared using the reaction sequence described in general procedure P, scheme 11, starting from intermediate 11b using 4-tert-butoxycarbonylacetylene (Aldrich) as the alkyne. By means of techniques known in the artWorking up the protected dicarboxylic acid 11e (R) by methods known to the skilled worker⁹' -tert-butyl propionate, P ═ Alloc), 4-methylthiazole moieties are attached. Deprotection of the ester as in general procedure P affords the desired carboxylate intermediate 11f (R)⁹' -4-methyl-thiazol-2-yl, P ═ Alloc).

4- [2- (4-methyl-thiazol-2-yl) -ethyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

2b (R) by means of the procedure described in example 26²' ═ H) with the protected amino acid 11f (R)⁹Coupling and deprotection of' -4-methyl-thiazol-2-yl, P ═ Alloc) gave the title lincosamide of example 28: (300MHz, CD) ₃OD)δ6.80(s，1)5.07(d，J＝5.5，1)，4.43(dd，J＝3.3，9.91)，3.91-3.86(m，2)，3.60(m，1)，3.50-3.45(m，1)，3.30(d，J＝3.3，10.4，1)，2.89-2.82(m，2)，2.76-2.67(m，1)，2.19(s，3)，1.96(s，3)，1.77-1.73(m，1)，1.61-1.57(m，3)，1.22-1.09(m，2)，0.72(d，J＝6.9，6)；MS(ESPOS)：488.4[M+H]⁺.

Example 29

4- (3-Methoxyimino-propan-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

11e(R⁹' -3-methoxyimino-propyl, P ═ Alloc).

To 11e (R) prepared in example 26⁹' -3-oxopropyl, P ═ Alloc) (129mg, 0.45mmol) in ethanol (1.3mL) was added methoxylamine hydrochloride and pyridine. The reaction mixture was refluxed for 2 hr. The solvent was removed under vacuum. The residue was dissolved in ethyl acetate, washed with 10% citric acid and brine, dried, and concentrated to give 11e (R)⁹' -3-methoxyimino-propyl, P ═ Alloc) (129mg, 91%) as a clear oil: ms (espos): 334.5[ M + Na ]]⁺.

4- (3-Methoxyimino-propyl) -piperidine-1, 2-dicarboxylic acid 1-allyl ester 11f (R)⁹' (3-methoxyimino-propyl), P ═ Alloc).

Ester 11e (R)⁹' (3-methoxyimino-propyl), P ═ Alloc) (129mg, 0.41mmol, 1equiv) mixture in THF (1.5mL) and water (0.5mL) was added lithium hydroxide monohydrate (69mg, 1.6mmol, 4 equiv). The mixture was stirred at rt overnight. THF was removed under vacuum. The aqueous layer was dissolved in ethyl acetate and partitioned with 10% citric acid. The organic layer was washed with water (1x) and brine (1x), dried, and concentrated to give 11f (R) ⁹' -3-methoxyimino-propyl, P ═ Alloc) (114mg, 93%) as a clear oil:

¹HNMR(300MHz，CDCl₃)δ7.31(t，J＝6.2，0.6H)，6.60(t，J＝5.4，0.4H)，5.96-5.82(m，1)，5.30-5.16(m，2)，4.58(d，J＝5.7，2)，4.83(t，J＝6.5，1)，3.83(d，J＝0.3，1.2H)，3.78(d，J＝0.6，1.8H)，3.77-3.62(m，1)，3.42-3.30(m，1)，2.38-2.26(m，1)，2.23-2.14(m，1)，2.08-1.85(m，2)，1.82-1.62(m，2)，1.53-1.35(m，3)；MS(ESPOS)：321.2[M+Na]⁺.

4- (3-methoxyimino-propyl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

At 0 ℃ to 2b (R)²' ═ H) (109.8mg, 0.38mmol, 1equiv) mixture in anhydrous DMF (0.9mL) triethylamine (0.26mL, 1.91mmol, 5equiv) was added, followed by the addition of bis (trimethylsilyl) trifluoroacetamide (0.15mL, 0.57mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 minutes and then at rt for 50 minutes. Adding the protected amino acid 11f (R) to the reaction mixture⁹' -3-methoxyimino-propyl, P ═ Alloc) (113.6mg, 0.38mmol, 1.0equiv) and HATU (182mg, 0.48 mm)ol, 1.26 equiv). The reaction mixture was stirred at rt for 3 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate and washed with 10% citric acid (1 ×), water (1 ×), sat₃(1x) and brine wash. Subjecting the organic layer to Na₂SO₄Drying and concentrating. The residue was purified by chromatography to give Alloc-protected lincosamide product (107mg, 53%): ms (espos): 532.4[ M + H]⁺.

To a mixture of the above Alloc-protected lincosamide (107mg, 0.20mmol, 1equiv) in THF (2.6mL) was added dimedone (282mg, 2.01mmol, 10equiv) and tetrakis (triphenylphosphine) palladium (46.5mg, 0.04mmol, 0.2 equiv). The mixture was stirred at rt overnight. The solvent was removed under vacuum and the residue was purified by chromatography to give the title compound of example 29 (28mg, 31%) as a white solid:

¹H NMR(300MHz，CD₃OD)δ7.36(t，J＝6.2，0.68H)，6.66(t，J＝5.4，0.32H)，5.23(d，J＝5.7，1)，4.16(dd，J＝3.2，10.1，1)，4.10-4.01(m，2)，3.81(s，1)，3.79(d，J＝3.3，1)，3.74(s，2)，3.50(dd，J＝3.3，9.9，1)，3.30-3.22(m，1)，3.18-3.10(m，1)，2.67-2.55(m，1)，2.40-2.31(m，1)，2.25--2.12(m，2)，2.10(s，3)，1.97-1.88(m，1)，1.76-1.65(m，1)，1.59-1.38(m，3)，1.14-1.01(m，2)，0.90(d，J＝6.9，6)；MS(ESPOS)：448.4[M+H]⁺.

Example 30

4- (3-ethoxyimino-propan-1-yl) -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 30 the title compound was synthesized as described in example 29, from intermediate 11e (R)⁹' -3-oxopropyl, P ═ Alloc) and starting from imine formationReplacing ethoxyamine hydrochloride in the step: ms (espos): 462.4[ M + H]⁺.

Example 31

4- [2- (5-ethyl-isoxazol-3-yl) -ethyl ] -piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

Example 31 the title compound was synthesized as described in example 29 from intermediate 11e (R)⁹Starting with' 3-oxopropyl, P ═ Alloc), replacement of hydroxylamine hydrochloride in the imine formation step provided 11e (R)⁹' -3-hydroxyimino-propyl, P ═ Alloc). Intermediate 11e (R) was processed by cycloaddition of 1-butyne in the presence of N-chlorosuccinimide and TEA⁹' -3-hydroxyimino-propyl, P ═ Alloc), installing an isoxazole heterocycle. Coupling and deprotection were performed as described in example 29: ms (espos): 486.3[ M + H]⁺.

Example 32

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4-position stereoisomer I and 4-position stereoisomer II

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide (high Rf 4-position stereoisomer I and low Rf 4-position stereoisomer II)

At 0 deg.C, to the general procedure described in scheme 12 of general procedure QBoc-protected amino acid 12d (P ═ Boc, R) prepared in sequence⁹To a solution of (310mg, 1.15mmol) in DMF (3mL) was added 7-Cl MTL 6b (R ═ propyl, m ═ 1) (R ═ m ═ 1)²＝H，R³Cl) (306mg, 1.15mmol), HBTU (469mg, 1.3mmol) and DIEA (290 μ L, 2.3mmol) were stirred at room temperature overnight. DMF was removed by rotary evaporation under high vacuum. The resulting residue was purified by silica gel column chromatography (3% MeOH in DCM) to give the desired Boc-protected 4-F lincosamide (451mg, 75%) as a light brown oil:

¹H NMR(300MHz，CD₃OD)δ5.29(d，J＝5.7，1)，4.50(m，3)，4.10(m，1)，3.60(m，3)，2.51(m，1)，2.11(m，4)，1.70(m，2)，1.50(m，9)，0.96(t，J＝7.2，3)；MS(ESPOS)：529[M+H]+.

to a solution of the Boc-protected 4-fluorolincosamide (451mg, 0.85mmol) in DCE (6mL) was added triethylsilane (0.16mL), TFA (2mL) and water (0.16mL), and the mixture was stirred at room temperature for 1.5 hr. The reaction solvent was removed in vacuo. The resulting residue was purified by column chromatography on silica gel using 10% MeOH in DCM as eluent to give the title compound of example 32, 4-stereoisomer I (high TLC Rf) (165mg, 45%):

¹H NMR(300MHz，CD₃OD)δ5.29(d，J＝5.7，1)，4.59(m，2)，4.32(d，J＝9.9，1)，4.07(dd，J＝5.7，10.2，1)，3.81(d，J＝3.3，1)，3.59(m，3)，3.01(d，J＝3.0，1)，2.83(m，1)，2.14(m，4)，1.86(m，2)，1.50(m，5)，0.99(t，J＝7.2，3)；MS(ESPOS)429[M+H]+；

And 4-stereoisomer II (low TLC Rf) (165mg, 45%):

¹H NMR(300MHz，CD₃OD)δ5.29(d，J＝5.7，1)，4.59(m，3)，4.29(d，J＝10.2，1)，4.08(dd，J＝5.7，10.2，1)，3.85(d，J＝3.3，1)，3.59(m，4)，2.60(m，1)，2.11(m，3)，1.88(m，2)，1.50(m，5)，0.99(t，J＝7.2，3)；MS(ESPOS)；429[M+H]+.

example 33

4-fluoro-4-propyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4-fluoro-4-propyl-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 12d (P ═ Boc, R)⁹N-propyl, m 2).

Synthesis of Boc-protected 4-fluoroamino acid 12d from starting material (2S) -4-oxo-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester general procedure Q as depicted in scheme 12 was used. The starting material (2S) -4-oxo-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester is prepared e.g. from Bousquet, y.; anderson, p.c.; bogri, t.; duceppe J.; grenier, l.; guse, i.; tetrahedron, 1997, 5315671-15680.

Rapidly stirring 1-tert-butyl 4-oxo-piperidine-1, 2-dicarboxylate 12a (m ═ 2, P ═ H, P₂Boc) (16.0g, 0.066mol) (prepared by the method described by Bousquet et al tetrahedron, 1997, 53, 15671) was treated with solid cesium carbonate (10.7g, 0.033mol) and methyl iodide (4.5mL, 0.072 mol). The reaction mixture was stirred for 5h, diluted with EtOAc, extracted with saturated aq. sodium bicarbonate, 10% aq. citric acid and brine, the organic layer was separated, dried over sodium sulfate, filtered and evaporated to dryness. Evaporation from dry benzene and azeotropic drying of the product from solvent removal gave 14.8g (98%) of the desired product, 1-tert-butyl 4-oxo-piperidine-1, 2-dicarboxylic acid 2-methyl ester 12a (m ═ 2, P ═ Me, P ₂Boc), as an oil: TLC Rf 0.53 (Hexane/Et OAc, 1: 1);¹H NMR(300MHz，CDCl₃) δ 5.33 (width m, 0.5) rotamer, 5.06 (width m, 0.5) rotamer, 4.31-4.19(m, 1), 3.95(s, 3), 3.95-3.70(m, 1), 3.16-2.97(m, 2), 2.71(m, 2), 1.68 (width s, 9).

Stirring the mixture at 0 deg.C to obtain 4-oxo-piperidine-1, 2-diCarboxylic acid 1-tert-butyl ester 2-methyl ester 12a (m ═ 2, P ═ Me₂Boc) (5.17g, 0.02mol) in DCM (60mL) was treated with tetraallyltin (Aldrich) (5.3mL, 0.022mol), followed by the dropwise addition of BF₃·OEt₂(2.5mL, 0.02 mol). The reaction mixture was stirred for 1h, then aq.1M potassium fluoride (38.0mL) and celite (5g) were added and the reaction mixture was stirred for 3 h. The reaction mixture was filtered, concentrated to dryness, the residue was dissolved in DCM, washed with water and brine, over MgSO₄Drying and evaporating to dryness. The resulting residue was purified by silica gel column chromatography (DCM 100% to DCM: acetone 9: 1) to give 3.85g (64%) of the desired product 4-allyl-4-hydroxy-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester 12b (m ═ 2, P ═ Me₂＝Boc，R⁹' -allyl) is an oil.¹H NMR(300MHz，CDCl₃) δ 6.11-5.97(m, 1), 5.42-5.32(m, 2), 5.06 (width d, J ═ 6.0, 0.5) rotamer, 4.87 (width d, J ═ 6.0, 0.5) rotamer, 4.18-4.03(m, 1), 3.93(s, 3), 2.48-2.37(m, 2), 1.98-1.43(m, 11); ms (espos): 322.0[ M + Na ] ]⁺.

A rotamer: rotamers

Stirring 12b (m-2, P-Me, P)₂＝Boc，R⁹' -allyl) (3.80mL, 1.27mmol) was stirred with a suspension of 10% Pd/C (degussa wet 50% w/w) (1.35g, 1.3mmol) in MeOH (80mL) under 1atm hydrogen for 6 h. The reaction mixture was filtered through celite, evaporated to dryness and azeotropically dried by evaporation from toluene and the resulting residue (3.15g) was used in the next step without further purification.

To a stirred-78 deg.C solution of DAST (1.7mL, 1.3mmol) in DCM (50mL) was added a solution of 4-hydroxy-4-propyl-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester in DCM (30 mL). The reaction mixture was then stirred at-78 ℃ for 1h and then warmed to-40 ℃ for 5 h. Additional DAST (0.4mL) was added, the reaction mixture was stirred for an additional 2h, and saturated aq.K was added₂CO₃(20mL) and water (60mL), followed by diethyl ether (500mL), the organic layer was separated, washed with brine, dried over sodium sulfate, and evaporated to dryness. Subjecting the crude fluorinated product to silica gel column chromatographyPurification (Hexane-EtOAc 9: 1). The residue from the chromatographic purification was dissolved in dioxane (65mL) and water (26mL), cooled to 0 ℃ and washed with OsO₄(0.65mL, 4% aqueous solution) and 30% H₂O₂(10 mL). The reaction mixture was stirred overnight, concentrated to dryness, the residue dissolved in DCM and the organic layer washed with water (100mL), 25% aq ₂SO₃(2X 100mL) and brine (100mL) over Na₂SO₄Drying and evaporating to dryness. The resulting residue was purified by silica gel column chromatography (hexane-EtOAc 9: 1) to give (1.08g, 34%) the desired product 4-fluoro-4-propyl-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester 12c (m ═ 2, P ═ Me, P ═ m₂＝Boc，R⁹N-propyl) is an oil.

¹HNMR(300MHz，CDCl₃)δ4.59(dd，J＝6.0，6.0，1)，3.82-3.69(m，1)，3.74(s，3)，3.28(m，1)，3.29-2.04(m，2)，1.91-1.71(m，3)，1.60-1.31(m，6)，1.45(s，9)，0.92(t，J＝7.1，3)；MS(ESPOS)：204.1(M+H-Boc)，326.3[M+Na]⁺.

Coupling from 12d (P ═ Boc, R) was readily accomplished using the coupling and deprotection conditions of example 32⁹N-propyl, m-2) the title compound of example 33 was synthesized.

MS(ESPOS)：443.1[M+H]⁺(ii) a HPLC: c183.5 μm, 4.6X 30mm column; gradient eluent 2% -98% MeCN over 10 min; 1.5 mL/min): rt 3.738min.

Example 34

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-hydroxy-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

12d (P ═ Boc, R) prepared via general procedure Q depicted in scheme 12⁹＝C₃H₇A stirred suspension of m-1) (164mg, 0.57mmol) was suspended in anhydrous acetonitrile (4 mL). Triethylamine (332. mu.L, 3.02mmol) was added and the reaction mixture was cooled to 0 ℃. Isobutyl chloroformate (78. mu.L, 0.57mmol) was added and after 10min the reaction was allowed to warm to 4 ℃. After 1.5h, a solution of MTL 1a (151mg, 0.57mmol) in 1: 1 acetone: water (4mL) was added and the reaction mixture was stirred at rt for 10 h. The reaction mixture was evaporated to dryness and chromatographed on silica (95: 5 dichloromethane/MeOH to 95: 8 dichloromethane/MeOH) to give the product as a colourless oil (137mg, 45%): TLC Rf ═ 0.32 (9: 1 dichloromethane/MeOH); ms (espos): 411[ M + H-Boc ]⁺，511[M+H]⁺.

To a solution of the Boc-protected lincosamide (125mg) above in DCM (2.0mL) was added DCE solution (10.0mL), trifluoroacetic acid (5mL), methyl sulfide (0.3mL) and water (0.3 mL). The reaction mixture was stirred at rt for 40min, then diluted with DCE (25.0 mL). The solvent was removed under vacuum and co-evaporated twice with DCE. The residue was chromatographed on fluorosil (20% MeOH0.25M NH)₃DCM) to give the title compound as a colourless solid (30.0mg, 30%): ms (espos): 411.6[ M + H]⁺.

Example 35

4-fluoro-4-butyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4-position stereoisomer I and 4-position stereoisomer II

4-hydroxy-4-butylproline methyl ester 12b (P ═ Boc, m ═ 1, R⁹N-butyl).

At-78 deg.C to n-butylLithium (165mg, 2.6mmol) in THF (5mL) was added with 12a (P ═ Boc, P) with stirring₂Me, m-1) (570mg, 2.3mmol) in THF (5 mL). The reaction mixture was stirred at-78 deg.C for 2hr, then at-40 deg.C for an additional 1 hr. EtOAc (20mL) was added followed by NH₄Cl (5mL, 10%) and water (10 mL). The organic layer was separated, dried over sodium sulfate and evaporated to dryness. The resulting residue was purified by silica gel column chromatography using 50% EtOAc in hexanes as the eluent to give 4-hydroxy-4-butylproline methyl ester 12b (P ═ Boc, m ═ 1, R) ⁹N-butyl) as a colorless oil (0.52g, 73%):

¹HNMR(300MHz，CDCl₃)δ4.33(m，1)，3.76(d，J＝4.8，3)，3.62(m，2)，3.28(m，1)，2.13(m，1)，2.02(m，1)，1.57(m，2)，1.29(m，12)，0.88(m，3)；MS(ESPOS)：324[M+Na]+.

4-fluoro-4-butylproline methyl ester 12c (P ═ Boc, m ═ 1, R⁹N-butyl).

To a stirred solution of DAST (0.55g, 3.4mmol) in DCM (5mL) was slowly added 4-hydroxyproline 12b (P ═ Boc, m ═ 1, R) at-78 ℃⁹N-butyl) (520mg, 1.7mmol) in anhydrous DCM (5 mL). The mixture was stirred at-78 ℃ for 1h, then at-10 ℃ for another 1 h. DCM (50mL) was added followed by aq₄Cl (10%, 30 mL). The organic layer was separated, dried over sodium sulfate and evaporated to dryness. The resulting residue was purified by column chromatography eluting with 5% EtOAc in hexanes to give 4-fluoro-4-butylproline methyl ester 12c (P ═ Boc, m ═ 1, R⁹N-butyl) (270mg, 52%) as a colorless oil:

¹HNMR(300MHz，CDCl₃)δ4.41(m，2)，3.83(m，1)，3.71(s，3)，3.45(dd，J＝12.3，32.7，2)，2.48(m，1)，1.73(m，2)，1.40(m，12)，0.89(m，3)；MS(ESPOS)：326[M+Na]⁺.

4-fluoro-4-butylproline 12d (P ═ Boc, m ═ 1, R⁹N-butyl).

To a solution of 12c (0.27g, 0.89mmol) in THF (10mL) and water (3mL) was added lithium hydroxide monohydrate (45mg, 1.06 mmol). The reaction mixture was stirred at room temperature overnight. THF was removed and the residue was purified by column chromatography using 10% MeOH in DCM as eluent to give 4-fluoro-4-butylproline 12d (P ═ Boc, m ═ 1, R⁹N-butyl) (0.26g, 100%) as a colorless oil: ¹H NMR(300MHz，CD₃OD)δ4.30(m，1)，3.72(m，1)，3.49(m，1)，3.39(m，1)，2.58(m，1)，2.02(m，2)，1.72(m，13)，0.93(t，J＝6.6，3)；MS(ESNEG)：288[M-1]^-.

4-fluoro-4-butyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide: 4-stereoisomer I and 4-stereoisomer II

At 0 deg.C, add 12d (P ═ Boc, m ═ 1, R)⁹N-butyl) (125mg, 0.43mmol) in DMF (3mL) was added 7-Cl MTL 6b (R)²＝H，R³Cl) (117mg, 0.43mmol), HBTU (180mg, 0.47mmol) and DIEA (111mg, 0.86mmol), and stirred at room temperature overnight. The solvent was removed and the residue was purified by silica gel column chromatography, eluting with 2% MeOH in DCM, to give the desired Boc-protected lincosamide intermediate (170mg, 72%) as a light brown liquid:¹HNMR(300MHz，CD₃OD)δ5.29(d，J＝5.4，1)，4.57(m，3)，4.39(m，1)，4.03(m，2)，3.74(m，3)，3.25(m，1)，2.51(m，1)，2.12(m，3)，1.85(m，3)，1.46(s，9)，1.36(m，6)，0.93(t，J＝6.6，3)；MS(ESPOS)：543[M+H]⁺.

to a solution of the Boc-protected lincosamide (170mg, 0.31mmol) in DCE (6mL) was added triethylsilane (0.16mL), TFA (2mL) and water (0.16 mL). The reaction mixture was stirred at room temperature for 1 hr. The solvent was removed and the residue was purified by silica gel column chromatography eluting with 10% MeOH in DCM to give 4-fluoro-4-butyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]Amide, 4-stereoisomer I (14mg, 10%):¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝6.0，1)，4.54(m，3)，4.29(d，J＝10.2，1)，4.09(dd，J＝5.6，10.2，1)，3.80

4-fluoro-4-butyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide, 4-stereoisomer II (3mg, 2%):

¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝6.0，1)，4.54(m，3)，4.29(d，J＝10.2，1)，4.09(dd，J＝5.6，10.2，1)，3.80(d，J＝3.0，1)，3.56(m，3)，2.70(m，1)，2.14(m，4)，1.87(m，2)，1.43(m，7)，0.94(t，J＝7.2，3)；(ESPOS)：443[M+H]⁺.

Example 36

4-fluoro-4-ethyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4-fluoro-4-ethyl-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 12d (P ═ Boc, m ═ 2, R⁹N-ethyl).

Synthesis of Boc-protected 4-fluoroamino acid 12d from starting (2S) -4-oxo-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester Using general procedure Q as depicted in scheme 12, trimethylsilyl acetylene anion was used as the two-carbon synthon in the 4-keto alkylation step. The starting 1-tert-butyl 4-oxo-piperidine-1, 2-dicarboxylate is prepared e.g. as Bousquet, y.; anderson, p.c.; bogri, t.; duceppe J.; grenier, l.; guse, i.; tetrahedron, 1997, 5315671-15680. Coupling from 12d (P ═ Boc, m ═ 2, R) was readily accomplished using the coupling and deprotection conditions of example 34⁹Ethyl) the title compound of example 36 was synthesized. Ms (espos): 429.1[ M + H]⁺.

Example 37

4-propylidene-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

4-oxo-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester.

To a solution of (2S) -4-oxo-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester (0.52g, 2.15mmol) in methanol (10mL) was added a 2M solution of TMS-diazomethane in hexane (2mL, 4mmol) and stirred at room temperature for 15 min. The reaction solvent was removed and the resulting methyl ester product (4-oxo-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester) was used directly in the next reaction (0.55g, 100%):

MS(ESPOS)：258[M+H]⁺；¹HNMR(300MHz，CD₃OD)δ5.13，4.86(bs，1)，4.02-4.11(m，1)，3.73(s，3)，3.67-3.72(m，1)，2.78(d，J＝4.2Hz，2)，2.51(bs，2)，1.46(bs，9).

4-propylene-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester.

A solution of propyltriphenylphosphonium bromide (1.24g, 3.22mmol) in THF (10mL) was added to a solution of sodium hydride (123mg, 3.22mmol) washed with hexane in THF (10mL) and stirred at room temperature for 3 h. To the above reaction mixture was slowly added methyl ester 12a (P ═ Boc, m ═ 2, P ═ 2₂Me) (0.55g, 2.15mmol) in THF (5mL) and then stirred for an additional 2 h. Then poured into water and extracted with ethyl acetate (30 mL). The organic phase was dried over magnesium sulfate, filtered and evaporated to dryness and the resulting residue was chromatographed, eluting with 20% EtOAc in hexanes to give the product 4-propylene-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (0.100g, 16%):

MS(ESPOS)：284[M+H]+；¹H NMR(300MHz，CD₃OD)δ5.18(m，m)，4.60-4.93(m，1)，3.91(m，1)，3.67(s，3)，2.94-3.04(m，2)，2.40-2.48(m，2)，1.99-2.06(m，1)，1.85-1.97(m，2)，1.38(bs，9)，0.85(t，J＝5Hz，3).

4-propylene-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester.

To a solution of 4-propylene-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (0.100g, 0.353mmol) in THF (10mL) was added a solution of lithium hydroxide (0.50g, 11.6mmol) in water (2mL) and the reaction mixture was stirred at room temperature for 16 h. Then poured into water and extracted with ether (20 mL). The aqueous layer was then acidified with 10% HCl (5mL) and extracted with ethyl acetate (30 mL). The product carboxylic acid 4-propylene-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester obtained after drying and removal of the solvent was used directly in the next step. Ms (esneg): 268[ M-1 ] ]-；¹HNMR(300MHz，CD₃OD)δ5.18(m，m)，4.60-4.93(m，1)，3.91(m，1)，2.94-3.04(m，2)，2.40-2.48(m，2)，1.99-2.06(m，1)，1.85-1.97(m，2)，1.38(bs，9)，0.85(t，J＝5Hz，3).

4-propylidene-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

To the stirred 2b (R)²' -H) (32mg, 0.12mmol), DIEA (0.2mL, 1.20mmol), 4-propylene-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester (37mg, 0.14mmol) in DMF (5mL) was added HBTU (57mg, 0.16mmol), and the mixture was stirred at room temperature for 2H. Most of the DMF was removed under high vacuum and the crude product was dissolved in ethyl acetate (50mL) and washed with saturated sodium bicarbonate (10 mL). The solvent was removed under vacuum and the product was purified by silica gel column chromatography using ethyl acetate as eluent to afford the desired Boc-protected lincosamide (50mg, 83%): ms (espos): 503 (M-1);¹H NMR(300MHz，CD₃OD)δ5.19-5.27(m，2)，4.10-4.26(m，2)，3.90-4.05(m，2)，3.83-3.90(m，2)，3.49-3.88(m，2)，3.06(m，2)，2.57(m，2)，1.90(s，3)，1.47(bs，9)，0.88-0.95(m，9).

to a solution of the Boc-protected lincosamide (50mg, 0.10mmol) in dichloroethane (6mL) was added triethylsilane (0.15mL), followed by 93% aq. trifluoroacetic acid (2.15 mL). After stirring at r t for 1hr, the solvent was removed under reduced pressure at 45 ℃. The resulting crude product was purified by column chromatography on silica gel using 10% MeOH in DCM as eluent to give the title compound (5mg, 12%):¹H NMR(300MHz，CD₃OD)δ5.40(m，1)，5.24(d，J＝3.8Hz，1)，4.16-4.20(m，1)，4.04-4.09(m，2)，3.81(t，J＝2.2Hz，1)，3.49-3.55(m，2)，2.50-2.92(m，3)，2.00-2.25(m，6)，2.10(s，3)，0.89-0.98(m，9).MS(ESPOS)：403[M+H]+.

example 38

4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Boc-protected amino acids with 2b (R)²' ═ H) coupling

To Boc-amino acid 21k (R) prepared by general method S as depicted in scheme 21 at 23 deg.C⁹Is n-propyl, R^9b＝H，m＝1)(69mg，0.26mmol，1equiv)、2b(R²' -H) (74mg, 0.26mmol, 1equiv) and HBTU (107mg, 0.28, 1.1equiv) in DMF (2.5mL) was added N, N-diisopropylethylamine (89 μ L, 0.51mmol, 2 equiv). The reaction was stirred at 23 ℃ for 2.5h, then concentrated in vacuo to remove DMF.

The resulting residue was dissolved in ethyl acetate (70mL) and then diluted with 1: 1 brine: 10% aqueous citric acid (50mL), saturated aqueous NaHCO₃Washed (50mL), brine (30mL), and dried (MgSO)₄) Filtered and concentrated to give 107mg of the desired coupled product. The product was used without further purification in the final deprotection step:¹H NMR(300MHz，CD₃OD)δ5.39(br d，J＝14.4Hz，1H)，5.19(d，J＝5.7Hz，1H)，4.10-3.82(m，4H)，3.55-3.48(m，1H)，2.45(br s，2H)，2.05(s，3H)，2.04-1.94(m，2H)，1.47(s，9H)，1.46-1.37(m，2H)，0.96-0.83(m，9H)；MS(ESPOS)：503.3[M+H]+.

boc-deprotection gives 4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

To a solution of the Boc-carbamate lincosamide (35mg, 0.070mmol, 1equiv) in DCE (5.0mL) at 23 deg.C was added H₂O (0.20mL), followed by TFA (2.0 mL). The reaction was stirred at 23 ℃ for 30min, then treated with toluene (40mL), then concentrated to a volume of 10mL, then treated with a second portion of toluene (40mL), and concentrated to dryness. The crude product was purified via semi-preparative HPLC (Waters) HR C18, 6 μm particle size,pore size, 25mm diameter × 100mm length, H containing 5-60% acetonitrile₂O/0.1% AcOH over 30min at a flow rate of 20mL/min) to give 16mg of pure 4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid]-amide:

¹H NMR(300MHz，D₂O)δ5.51(brs，1H)，5.33(br d，J＝5.4Hz，1H)，4.18(s，2H)，4.13-3.98(m，2H)，3.86(br s，1H)，3.55-3.58(m，3H)，2.58-2.39(m，2H)，2.12(s，3H)，2.12-2.00(m，3H)，1.50-1.37(m，2H)，0.94-0.78(m，9H)；¹³C NMR(300MHz，D₂O)：δ170.9，136.5，114.5，88.4，70.9，69.3，68.8，68.2，55.1，53.0，42.3，38.3，29.9，27.7，20.0，19.9，14.7，13.3，13.1；MS(ESPOS)：403.3[M+H]+.

example 39

4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Boc-protected amino acids with 6b (R)²＝H，R³═ Cl) coupling

To Boc-amino acid 21k (R) prepared by general method S as depicted in scheme 21 at 23 deg.C⁹N-propyl, m-1) (131mg, 0.49mmol, 1equiv), 7-Cl MTL 6b (R²＝H，R³Cl) (132mg, 0.49mmol, 1equiv) and HBTU (203mg, 0.54mmol, 1.1equiv) in DMF (4.0mL) was added N, N-diisopropylethylamine (170 μ L, 0.97mmol, 2 equiv). The reaction was stirred at 23 ℃ for 2.5h, then concentrated in vacuo to remove DMF. The resulting residue was dissolved in EtOAc (70mL) and then treated with 1: 1 brine: 10% aqueous citric acid (50mL), saturated aqueous NaHC0₃(25mL), washed with brine (30mL), and dried (MgSO)₄) Filtered and concentrated to give 276mg of the desired conjugate product. The product was used without further purification in the final deprotection step:

¹H NMR(300MHz，CD₃OD)δ5.41(br s，1H)，5.28(d，J＝6.0Hz，1H)，4.65-4.52(m，1H)，4.46-4.36(m，1H)，4.25-4.16(m，1H)，4.15-3.97(m，2H)，3.93-3.74(m，2H)，3.55(dd，J＝3.3，10.2Hz，1H)，2.62-2.40(m，2H)，2.13(s，3H)，2.10-1.95(m，2H)，1.49(s，9H)，1.46-1.32(m，5H)，0.90(brt，J＝7.2Hz，3H)；MS(ESPOS)：523.2[M+H]+.

4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

To a solution of the Boc-carbamate protected lincosamide (225mg, 0.43mmol, 1equiv) in DCE (25mL) at 23 deg.C was added H₂O (1.0mL), followed by TFA (10 mL). The reaction was stirred at 23 ℃ for 30min and then with toluene (C.) (150mL) and then concentrated to a volume of 30mL, then treated with a second portion of toluene (150mL) and concentrated to dryness. The crude product was purified via semi-preparative HPLC (Waters)HR C18, 6 μm particle size,pore size, 25mm diameter × 100mm length, H containing 5-60% acetonitrile₂O/0.1% AcOH over 30min at a flow rate of 20mL/min) to give 90mg of pure 4-propyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid]-amide:¹H NMR(300MHz，D₂O)δ5.53(br s，1H)，5.39(br d，J＝6.0Hz，1H)，4.66-4.55(m，1H)，4.46(dd，J＝1.2，10.2Hz，1H)，4.33(d，J＝9.9Hz，1H)，4.19-4.07(m，2H)，3.88(d，J＝2.7Hz，1H)，3.74(br s，2H)，3.66(dd，J＝3.0，10.2Hz，1H)，2.70-2.44(m，2H)，2.18(s，3H)，2.08(br t，J＝7.2Hz，3H)，1.52-1.37(m，2H)，1.42(d，J＝6.9Hz，3H)，0.85(t，J＝6.9Hz，3H)；MS(ESPOS)：423.1[M+H]+.

example 40

1-carbamoylmethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

To a stirred solution of 4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide (200mg, 0.48mmol, 1equiv) in anhydrous acetonitrile (3mL) was added triethylamine (0.2mL, 1.44mmol, 3equiv) followed by bromoacetamide (80mg, 0.58mmol, 1.2equiv) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 18h and evaporated to dryness. The resulting residue was first purified on silica gel eluting with 7% ammonia in methanol/dichloromethane. The desired fractions were collected, evaporated to dryness and repurified by HPLC (removal of by-products of the reaction of the base with bromoacetamide). After lyophilization, the desired title compound of example 40 (2.0mg) was obtained as a white fluffy powder:

HPLC：R_t＝4.11min(220.0nm)；¹H NMR(300MHz，CD₃OD)δ(rotamers)5.46(d，J＝5.5，1)，4.47(dd，J＝3.02，2.7，1.1)，4.31-4.25(m，3.3)，4.11(d，J＝3.02，1.6)，2.31(s，3)，1.65-1.52(m，9.5)，1.12-1.09(m，10.3).MS(ESPOS)：476.5[M+H]+，(ESNEG)：474.5[M-H]-.

EXAMPLE 41

1-cyanomethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

A stirred solution of crude 4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide (210mg, 0.50mmol, 1equiv) and triethylamine (0.21mL, 1.51mmol, 3equiv) in dry acetonitrile (3mL) was treated with bromoacetonitrile (42 μ L, 0.60mmol, 1.2equiv) at rt and under nitrogen. The resulting reaction mixture was stirred at rt for 18h, evaporated to dryness and purified by chromatography on silica gel with 2.5% methanol in dichloromethane as eluent. The desired fractions were pooled, evaporated to dryness, and lyophilized to give the title compound (11.2mg, 10%) as a fluffy white powder:

¹H NMR(300MHz，CD₃OD)δ5.44(d，J＝5.49，1)，4.38-4.23(m，4)，2.29(s，3)，1.52(m，11)，1.16-1.09(m，12)；MS(ESPOS)：458.5[M+H]+，MS(ESNEG)：456.5[M-H]-.

example 42

1- (1H-imidazol-2-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pent-2-enyl-pyrrolidine-2-carboxylic acid benzyl ester.

Boc-protected amino acid 7d (R)⁹' -4-pent-2-enyl) (433mg, 1.16mmol, 1eq.) was stirred in 4M HCl in dioxane (5mL) for 2h and then evaporated to dryness. The resulting residue was co-evaporated to dryness (3 × 20mL) from DCM. The crude HCl salt was dissolved in acetone (8mL), and the resulting solution was treated with diisopropylethylamine (0.61mL, 3.50mmol, 3equiv), followed by 1-benzyl-2- (chloromethyl) -1H-imidazole (Maybridge) (338mg, 1.39mmol, 1.2 equiv). The reaction mixture was stirred at room temperature for 48h and evaporated to dryness. The resulting residue was diluted with EtOAc (200mL), washed sequentially with 10% citric acid, brine, and Na ₂SO₄Dried, filtered and evaporated to dryness. The crude product was purified by silica gel chromatography using CH₂Cl₂Hexane/MeOH (6: 5: 1) to give the desired N-alkylated product, benzyl 1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pent-2-enyl-pyrrolidine-2-carboxylate (257mg, 50%): rf is 0.7 (7: 2: 1, CH)₂Cl₂hexane/MeOH); ms (espos): 444.3[ M + H]⁺.

1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pent-2-enyl-pyrrolidine-2-carboxylic acid.

To a stirred benzyl 1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pent-2-enyl-pyrrolidine-2-carboxylate (257.2mg, 0.6mmol, 1equiv) in 4: 1THF/H at room temperature₂Lithium hydroxide monohydrate (250mg, 5.96mmol, 10equiv) was added to a solution of O (8 mL). The resulting reaction mixture was stirred at room temperature overnight and evaporated to dryness. Subjecting the obtained product toThe residue was dissolved in water (10mL), the pH of the resulting solution was adjusted between 3 and 4, and extracted with EtOAc (3X 100 mL). The combined organic extracts were washed with brine and then Na₂SO₄Dried, filtered and evaporated to dryness to give the acid product 1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pent-2-enyl-pyrrolidine-2-carboxylic acid (202mg, 98%): rf is 0.4 (7: 2: 1 CH)₂Cl₂hexane/MeOH), KMnO₄Visualization of the stain; ms (espos): 355[ M + H ]⁺；MS(ESNEG)：352[M-H]^-.

1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

In N₂And 2b (R) stirred at 0 deg.C²' ═ H) (96.4mg, 0.33mmol, 1equiv) in anhydrous DMF (1.5mL) was added triethylamine (0.4mL, 2.9mmol, 8.7equiv), followed by BSTFA (0.4mL, 1.51mmol, 4.6 equiv). The resulting mixture was stirred at 0 ℃ for 10min, then at room temperature for 30min, and then cooled. To the reaction was added a solution of 1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pent-2-enyl-pyrrolidine-2-carboxylic acid (194mg, 0.55mmol, 1.7equiv), followed by solid HATU (261mg, 0.69mmol, 2.1 equiv). The resulting mixture was stirred at room temperature for 3h and evaporated to dryness. The resulting residue was dissolved in EtOAc and then treated with 10% aqueous citric acid, saturated aqueous NaHCO₃Washed with brine and dried (MgSO)₄) Filtering, and concentrating. The crude per-silylated compound was dissolved in MeOH (60mL) and treated with Dowex H at room temperature⁺Resin (250mg) was treated for 45min, the reaction mixture was filtered and evaporated to dryness to give the lincosamide product 1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -2-carboxylic acid ]-an amide.

Oven-dried sealed tubes were charged with the crude 1- (1-benzyl-1H-imidazol-2-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -2-carboxylic acid]-amide (280mg, 0.48mmol) in anhydrous EtOH (5mL), 10% palladium on charcoalCarbon (560mg) and 1, 4-cyclohexadiene (1.5 mL). N for reaction vessel₂Purify, seal, and stir at room temperature for 18 h. The reaction mixture was filtered through celite, washed several times with reagent alcohol, the washings and filtrate were combined and evaporated to dryness. The residue obtained is chromatographed on silica gel (1: 9 ammonia in methanol: CH)₂Cl₂). The desired fractions were evaporated to dryness and lyophilized to give the title compound of example 42 (29.3mg, 18%): TLC Rf 0.7 (14% ammonia in methanol/CH)₂Cl₂) KMnO4 visualization stain; ms (espos): 499.4[ M + H]⁺

Example 43

1-aminomethyl-4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Stirring 4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]A suspension of amide (153mg, 0.34mmol, 1equiv) and ethyl imidoformate (ethyl formimidate) (Aldrich) (44mg, 0.40mmol, 1.2equiv) in dioxane (1mL) was treated with 1M aqueous NaOH (0.74mL, 0.74mmol, 2.2 equiv). After 1h, additional ethyl imidate (44mg, 0.40mmol, 1.2equiv) was added to the reaction mixture and stirring was continued for 30 min. The reaction mixture was frozen and lyophilized. The lyophilized powder was purified by column chromatography to give the title compound (8 mg): TLC Rf ═ 0.48CHCl ₃MeOH/32% aqueous AcOH (5: 3: 1); ms (espos): 447.7[ M + H]⁺，469.7[M+Na]⁺，MS(ESNEG)：481.6[M-H+HCl]^-.

Example 44

4-butyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The Boc-amino acid 21k (R) prepared with the aid of general method S⁹N-butyl, R^9bH, m-1) with 7-Cl MTL 6b (R)²＝H，R³═ Cl) coupling. The product was used in the final deprotection step without further purification. Deprotection and purification as before in example 38 gave the title compound.

¹H NMR(300MHz，CD₃OD)δ5.52(br s，1H)，5.29(br d，J＝5.7Hz，1H)，4.63-4.52(m，2H)，4.30(d，J＝9.6Hz，1H)，4.08(dd，J＝5.7Hz，1H)，4.00(dd，J＝4.8，11.4Hz，1H)，3.81(d，J＝2.1Hz，1H)，3.69(brs，2H)，3.56(dd，J＝3.3，10.2Hz，1H)，2.66-2.35(m，2H)，2.20-2.06(m，2H)，2.14(s，3H)，1.54-1.28(m，7H)，0.93(t，J＝7.2Hz，3H)；MS(ESPOS)：437.2[M+H]⁺.

Example 45

4-butyl-1, 2, 3, 6-tetrahydro-pyridine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The Boc-amino acid 21k (R) prepared with the aid of general method S⁹N-butyl, R^9bH, m-1) and lincosamine 2b (R)²' ═ H) coupling. The product was used in the final deprotection step without further purification. Deprotection and purification as before in example 38 gave the title compound.¹H NMR(300MHz，D₂O)δ5.52(br s，1H)，5.24(d，J＝5.7Hz，1H)，4.23(dd，J＝3.6，10.2Hz，1H)，4.13-4.04(m，2H)，3.95(dd，J＝5.1，11.1Hz，1H)，3.81(d，J＝2.7Hz，1H)，3.68(bs，2H)，3.51(dd，J＝3.3，10.2Hz，1H)，2.59-2.34(m，2H)，2.24-2.06(m，3H)，2.11(s，3H)，1.52-1.28(m，4H)，0.98-0.87(m，9H)MS(ESPOS)：417.3[M+H].

Example 46

5-propyl-2, 3, 6, 7-tetrahydro-1H-aza-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]-amides of

At 0 deg.C, to the protected cyclic amino acid 22h (R) ^9b＝H，R⁹To a solution of 340mg (0.88 mmol) in DMF (3mL) was added 6b 7-Cl MTL (R)²＝H，R³Cl) (367mg, 0.88mmol), TEA (332 μ L, 1.76mmol), HRTU (496mg, 0.97mmol), and stirred at room temperature overnight. The solvent is then removed. Purification by column chromatography on silica eluting with 50-100% EtOAc in hexanes afforded the desired protected lincosamide 13a (R)⁹Is propyl radical, R^9b＝H，m＝2，R²＝H，R³＝Cl，P¹＝H，P²＝Boc)(575mg，90％)。MS(ESPOS)：537[M+1]⁺.

To Boc-protected lincosamide 13a (R)⁹Is propyl radical, R^9b＝H，m＝2，R²＝H，R³＝Cl，P¹＝H，P²Boc) (575mg, 1.07mmol) in DCE (15mL) was added triethylsilane (0.5mL), TFA (5mL), and water (0.5mL), and stirred at room temperature for 1.5 hr. The reaction solvent was removed and the resulting residue was chromatographed on silica, eluting with 5-10% MeOH/DCM to give the title compound (433mg, 92%) as a colorless solid. Ms (espos): 437[ M +1 ]]⁺.

Example 47

5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The unsaturated title compound 5-propyl-2, 3, 6, 7-tetrahydro-1H-aza from example 46-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]Amide (433mg, 0.99mmol) and 10% Pd/C (80mg) in MeOH (10mL) was hydrogenated at 50psi overnight. The solvent was removed to give the crude product. Purification by silica gel column chromatography (20% MeOH/DCM) followed by preparative HPLC (general procedure AC) gave isomer 1Rt ═ 18.3min (10mg, 2.3%) and isomer 2Rt ═ 18.6min (58mg, 13.3%).

Isomer 1¹HNMR(300MHz，CD₃OD)δ5.29(d，J＝5.4，1)，4.88(m，1)，4.42(dd，J＝1.8，10.2，1)，4.23(d，J＝9.9，1)，4.10(dd，J＝5.7，10.2，1)，3.78(d，J＝3.3，1)，3.71(t，J＝6.0，1)，3.58(dd，J＝3.3，10.2，1)，3.15(m，1)，2.83(m，1)，2.13(m，2)，2.03(m，2)，1.86(m，2)，1.73(m，1)，1.50(m，4)，1.42(m，6)，0.92(t，J＝6.6，3)；MS(ESPOS)：439[M+H]⁺.

Isomer 2¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝5.7，1)，5.30(d，J＝5.7，1)，4.61(m，2)，4.29(d，J＝9.9，1)，4.10(dd，J＝5.7，10.2，1)，4.00(m，1)，3.78(d，J＝3.0，1)，3.58(dd，J＝3.3，10.2，1)，3.39(m，1)，3.08(m，1)，2.14(m，4)，1.96(m，3)，1.59(m，3)，1.45(m，3)，1.35(m，4)，0.93(t，J＝6.9，3)；MS(ESPOS)：439[M+H]⁺.

Example 48

1-cyclopropyl-5-propyl-azepane-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

To 5-propyl-azepane-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl, prepared by the methods described in examples 47, 48]-amide (68mg, 0.16mmol) in MeOH (2mL) acetic acid (0.1mL) followed by 1- [ (ethoxycyclopropyl) oxy]Trimethylsilane (0.2mL, 0.96mmol), sodium cyanoborohydride (41mg, 0.64mmol) andheating to reflux for 3 hr. The molecular sieve is filtered off and the reaction solvent is removed to obtain the crude product. Purification by column chromatography on silica gel (10% MeOH/DCM) and HPLC gave the title compound (44mg, 59%).¹H NMR(300MHz，CD₃OD)δ5.20(d，J＝5.4，1)，4.15(d，J＝6.6，1)，4.08(dd，J＝5.4，9.9，1)，3.96(d，J＝3.0，1)，3.88(t，J＝13.2，1)，3.62(m，1)，3.56(dd，J＝3.3，10.2，1)，3.14(m，1)，2.82(m，1)，2.13(m，2)，2.05(s，3)，1.99-1.30(m，10)，0.96(m，9)，0.51(m，4)；MS(ESPOS)：459[M+H]⁺.

Example 49

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by reacting isomer 1 or isomer 2 of the title compound 1 of example 46 (wherein R is R) in DMF in the presence of sodium carbonate²＝H，R³＝Cl，R⁶＝H，R⁹5-n-propyl, m-3) with 4-bromomethyl-5-methyl- [1, 3 ]Dioxol-2-ones (prepared as described in J.Alexander, et. al.J.Med.chem, 1996, 39, 480-.

Example 50

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] 5-propyl-azepane-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester

The title compound may be prepared by reacting isomer 1 or isomer 2 of the title compound 1 of example 46 (wherein R is R) in DMF in the presence of potassium bicarbonate²＝H，R³＝Cl，R⁶＝H，R⁹5-n-propyl, m ═ 3) with carbonic acid 5-methyl-2-oxo- [1, 3]Dioxol-4-ylmethyl ester 4-nitro-phenyl ester (prepared as described in F.Sakamoto, et. al, chem.Pharm.Bull.1984, 32 (6); 2241-.

Example 51

5-methyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Making lincosamine as described in general procedure Z6b 7-Cl MTL(R²＝H，R³═ Cl) with cyclic amino acid 22f (R) prepared by general method T⁹Is propyl radical, R^9bH, m ═ 2) coupling to give intermediate carbamate 13a (R)⁹Methyl, R^9b＝H，m＝2，R²＝H，R³＝Cl，P¹＝H，P²Boc), deprotection under acidic conditions yielded the crude unsaturated intermediate. Unsaturated Compounds in MeOH at 50psi H ₂Hydrogenation with 10% Pd/C gave a crude mixture of the 5-position isomer. The two 5-position isomers were separated by means of preparative HPLC.

Isomer 1 (Low Rt)¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝5.7，1)，4.58(dd，J＝6.3，10.8，2)，4.30(d，J＝9.9，1)，4.10(m，1)，3.79(m，1)，3.58(m，1)，3.33(m，1)，3.13(m，1)，2.14(m，4)，1.92(m，3)，1.55(m，1)，1.44(d，J＝9.9，6)，1.00(d，J＝9.9，3)；R_t：14.2min；MS(ESPOS)：412[M+H]⁺.

Isomer 2 (high Rt)¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝5.7，1)，4.59(m，2)，4.30(d，J＝9.9，1)，4.10(m，1)，3.99(m，1)，3.81(m，1)，3.58(m，1)，3.14(m，1)，2.14(m，4)，1.90(m，3)，1.51(m，1)，1.44(d，J＝9.9，6)，1.00(d，J＝9.9，3)；R_t＝14.5min；MS(ESPOS)：412[M+H]⁺.

Example 52

5-Ethyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Lincosamine 6b 7-Cl MTL (R) as described in general procedure Z²＝H，R³Ci) with a cyclic amino acid 22f prepared by general method T(R⁹Is propyl radical, R^9bH, m ═ 2) coupling to give intermediate carbamate 13a (R)⁹Methyl, R^9b＝H，m＝2，R²＝H，R³＝Cl，P¹＝H，P²Boc), deprotection under acidic conditions yielded the crude unsaturated intermediate. Unsaturated Compounds in MeOH at 50psi H₂Hydrogenation with 10% Pd/C gave a crude mixture of the 5-position isomer. The two 5-position isomers were separated by means of preparative HPLC.

Isomer 1 (Low Rt)¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝6.0，1)，4.59(m，1)，4.49(m，1)，4.27(d，J＝9.9，1)，4.10(dd，J＝6.0，10.2，1)，3.92(m，1)，3.79(m，1)，3.55(m，1)，2.99(m，1)，2.14(m，4)，1.79(m，3)，1.45(d，J＝9.9，6)，1.38(m，3)，0.99(m，3)；R_t＝14.6min MS(ESPOS)：425.3[M+H]⁺.

Isomer 2 (high Rt)¹HNMR(300MHz，CD₃OD)δ5.30(d，J＝6.0，1)，4.80(m，2)，4.29(d，J＝9.9，1)，4.10(dd，J＝6.0，10.2，1)，3.98(m，1)，3.80(d，J＝2.7，1)，3.59(dd，J＝3.0，10.2，1)，3.07(m，1)，2.14(m，4)，1.92(m，3)，1.52(m，1)，1.45(d，J＝9.9，6)，1.32(m，2)，0.93(m，3)R_t：16min；MS(ESPOS)：425.3[M+H]⁺.

Example 53

5-Cyclopropylmethyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹Is cyclopropylmethyl, R^9bH, m 2) and LinkeAmine 6b 7-Cl MTL (R) ²＝H，R³Cl). Hydrogenation of the unsaturated compound with 10% Pd/C as in example 47 gave the title compound as a mixture of 5-position isomers.

Example 54

5-cyclopropyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹Is cyclopropyl, R^9bH, m-2) with lincosamine 6b7-Cl MTL (R)²＝H，R³Cl). Hydrogenation of the unsaturated compound with 10% Pd/C as in example 47 gave the title compound as a mixture of 5-position isomers.

Example 55

5-Ethyl-4-methyl-azepan-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹Ethyl, R^9bMethyl, m 2) with lincosamine 6b7-Cl MTL (R)²＝H，R³Cl). Hydrogenation of the unsaturated compound with 10% Pd/C as in example 47 gave the title compound as a mixture of isomers.

Example 56

4-Ethyl-5-methyl-azepan-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹Methyl, R^9bEthyl, m 2) with lincosamine 6b 7-Cl MTL (R)²＝H，R³Cl). Hydrogenation of the unsaturated compound with 10% Pd/C as in example 47 gave the title compound as a mixture of isomers.

Example 57

5-Ethyl-6-methyl-azepan-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹Methyl, R^9bEthyl, m ═ 1) with lincosamine 6b 7-Cl MTL (R)²＝H，R³Cl). Hydrogenation of the unsaturated compound with 10% Pd/C as in example 47 gave the title compound as a mixture of isomers.

Example 58

4-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹＝H，R^9bPropyl, m 2) with lincosamine 6b 7-ClMTL (R)²＝H，R³Cl). Hydrogenation of the unsaturated compound with 10% Pd/C as in example 47 gave the title compound as a mixture of 4-positional isomers.

Example 59

5-fluoro-5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound may be prepared by coupling the cyclic amino acid 12d (R) as described in general procedure Z⁹Propyl, m 2) with lincosamine 6b 7-Cl MTL (R)²＝H，R³＝Cl)。

Example 60

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ cyclopropyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹Is cyclopropyl, R¹SMe) was coupled with 4-n-propylgulonic acid (4-n-propylhydyoglic acid) prepared by the method of hoeksme, h.et.al.journal of the American chemical society, 1967, 892448-2452 to give the title compound.

¹H MR(300MHz，D₂O)δ5.37(d，J＝5.7Hz，1H)，4.26-4.18(m，2H)，4.10(dd，J＝5.7，9.9Hz，1H)，3.98(d，J＝3.0Hz，1H)，3.82(dd，J＝6.6，11.1Hz，1H)，3.72(d，J＝8.7Hz，1H)，3.67(dd，J＝3.3，7.2Hz，1H)，2.96-2.83(m，1H)，2.90(s，3H)，2.45-2.16(m，3H)，2.10(s，3H)，1.50-1.22(m，4H)，1.10-0.98(m，1H)，0.86(t，J＝7.2Hz，3H)，0.67-0.56(m，1H)，0.50-0.40(m，1H)，0.32-0.14(m，2H).MS(ESPOS)：403.3[M+H]；MS(ESNEG)：437.2[M+Cl].

Example 61

4-propyl-piperidine-2-carboxylic acid [ cyclopropyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹Is cyclopropyl, R¹SMe) with 4-propyl-piperidine-1, 2-dicarboxylic acid-1-tert-butyl ester 27b (R)⁹Propyl) to yield the title compound. Reaction of intermediate 13a (R)¹＝SMe，R²⁰+R²¹Is cyclopropyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

¹HNMR(300MHz，D₂O)δ5.37(d，J＝5.7Hz，1H)，4.20(d，J＝9.0Hz，1H)，4.09(dd，J＝5.7，10.2Hz，1H)，3.96(d，J＝3.3Hz，1H)，3.85(dd，J＝3.0，12.9Hz，1H)，3.75-3.65(m，2H)，3.51-3.42(m，1H)，3.07-2.96(m，1H)，2.21-2.10(m，1H)，2.10(s，3H)，1.99-1.90(m，1H)，1.80-1.65(m，1H)，1.46-1.23(m，6H)，1.11-0.98(m，1H)，0.85(t，J＝6.6Hz，3H)，0.66-0.55(m，1H)，0.50-0.36(m，1H)，0.30-0.12(m，2H)；MS(ESPOS)：403.3[M+H]；MS(ESNEG)：437.2[M+Cl].

Example 62

5-propyl-azepane-2-carboxylic acid [ cyclopropyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Lincosamide 23g (R) as described in general coupling procedure Z²⁰+R²¹Is cyclopropyl, R¹SMe) with 5-propyl-azaCoupling of-1, 2-dicarboxylic acid-1-tert-butyl ester gives the title compound. Reaction of intermediate 13a (R)²⁰+R²¹Is cyclopropyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-3) under acidic conditions to give the title compound.

MS(ESPOS)：451.2[M+H]⁺.

Example 63

4-propyl-piperidine-2-carboxylic acid [ phenyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Preparation of lincosamide 23f (R) as illustrated in scheme 23²⁰+R²¹＝Ph，R¹＝SMe)。

¹H NMR(300MHz，CDCl₃)δ7.41-7.26(m，5H)，6.90(br d，J＝9.6Hz，1H)5.55(d，J＝5.4Hz，1H)，5.49(dd，J＝1.2，3.3Hz，1H)，5.30-5.23(m，2H)，5.17(dd，J＝3.3，10.8Hz，1H)，4.68(dd，J＝0.9，8.4Hz，1H)，2.08(s，3H)，2.06(s，3H)，1.97(s，3H)，1.62(s，3H)；MS(ESPOS)：530.0[M+Na]；MS(ESNEG)：506.0[M-H].

Preparation of lincosamide 23g (R) as described in scheme 23²⁰+R²¹＝Ph，R¹＝SMe)。

¹HNMR(300MHz，CD₃OD)δ7.40-7.20(m，5H)，5.09(d，J＝5.7Hz，1H)，4.16-4.05m，4H)，3.58(dd，J＝3.3，10.2Hz，1H)，1.38(s，3H)；MS(ESPOS)：286.0[M+H]；MS(ESNEG)：284.2[M-H].

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹＝Ph，R¹SMe) with 4-propyl-piperidine-1, 2-dicarboxylic acid-1-tert-butyl ester 27b (R)⁹Propyl) to yield the title compound. Reaction of intermediate 13a (R)²⁰+R²¹＝Ph，R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

¹H NMR(300MHz，D₂O)δ7.46-7.30(m，5H)，5.15(d，J＝5.7Hz，1H)，5.11(d，J＝9.9Hz，1H)，4.51(d，J＝10.2Hz，1H)，4.13-4.03(m，2H)，3.91(dd，J＝3.0，12.9Hz，1H)，3.68(dd，J＝3.3，10.2Hz，1H)，3.49-3.40(m，1H)，3.07-2.95(m，1H)，2.10-2.01(m，1H)，1.96-1.86(m，1H)，1.78-1.62(m，1H)，1.51(s，3H)，1.36-1.07(m，6H)，0.82(t，J＝6.6Hz，3H)；MS(ESPOS)：439.3[M+H]；MS(ESNEG)：473.2[M+Cl]

Example 64

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ phenyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Kindergarten as described in general procedure ZCoamide 23g (R) ²⁰+R²¹Is phenyl, R¹SMe) was coupled with 4-n-propyl gulonic acid to give the title compound.

¹HNMR(300MHz，D₂O)δ7.47-7.30(m，5H)，5.15(d，J＝5.7Hz，1H)，5.11(d，J＝10.2Hz，1H)，4.50(d，J＝9.9Hz，1H)，4.28(dd，J＝5.4，9.3Hz，1H)，4.11-4.04(m，2H)，3.75(dd，J＝6.0，11.1Hz，1H)，3.68(dd，J＝3.3，10.5Hz，1H)，2.91(s，3H)，2.90-2.80(m，1H)，2.45-1.90(m，3H)，1.48(s，3H)，1.44-1.10(m，4H)，0.78(t，J＝7.2Hz，3H)；MS(ESPOS)：439.3[M+H]；MS(ESNEG)：473.2[M+Cl].

Example 65

4-propyl-piperidine-2-carboxylic acid [ cyclopentyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Preparation of lincosamide 23f (R) as illustrated in scheme 23²⁰+R²¹Is cyclopentyl, R¹＝SMe)。

¹H NMR(300MHz，CDCl₃)δ6.29(brd，J＝9.0Hz，1H)，5.63(d，J＝5.7Hz，1H)，5.43(d，J＝3.3Hz，1H)，5.25(dd，J＝5.4，11.1Hz，1H)，5.14(dd，J＝3.3，11.1Hz，1H)，4.40-4.38(m，2H)，2.30-2.08(m，1H)，2.14(s，3H)，2.09(s，3H)，2.08(s，3H)，1.97(s，3H)，1.86-1.48(m，6H)，1.27-1.12(m，2H)；MS(ESPOS)：522.2[M+Na]；MS(ESNEG)：498.2[M-H].

Preparation of lincosamide 23g (R) as described in scheme 23²⁰+R²¹Is cyclopentyl, R¹＝SMe)。

¹H NMR(300MHz，CD₃OD)δ5.28(d，J＝5.7Hz，1H)，4.16-4.08(m，2H)，3.93(dd，J＝0.9，6.6Hz，1H)，3.54(dd，J＝3.0，10.2Hz，1H)，2.99(t，J＝6.6Hz，1H)，2.17-2.04(m，1H)，2.07(s，3H)，1.88-1.51(m，6H)，1.42-1.26(m，2H)

MS(ESPOS)：278.3[M+H]；MS(ESNEG)：276.2[M-H].

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹Is cyclopentyl, R¹SMe) with 4-propyl-piperidine-1, 2-dicarboxylic acid-1-tert-butyl ester 27b (R)⁹Propyl) to yield the title compound. Reaction of intermediate 13a (R)²⁰+R²¹Is cyclopentyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

¹H MR(300MHz，D₂O)δ5.37(d，J＝5.7Hz，1H)，4.23(dd，J＝5.4，9.3Hz，1H)，4.15(d，J＝9.3Hz，1H)，4.09(dd，J＝5.7，10.5Hz，1H)，3.90(dd，J＝3.3，13.2Hz，1H)，3.63(dd，J＝3.3，10.5Hz，1H)，3.49(br d，J＝12.6Hz，1H)，3.10-2.98(m，1H)，2.32-2.12(m，2H)，2.13(s，3H)，2.10-1.91(m，1H)，1.80-1.04(m，16H)，0.87(t，J＝6.6Hz，3H)；MS(ESPOS)：431.3[M+H]；MS(ESNEG)：465.2[M+Cl].

Example 66

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ cyclopentyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹Is cyclopentyl, R¹SMe) was coupled with 4-n-propyl gulonic acid to give the title compound.

¹H NMR(300MHz，D₂O)δ5.35(d，J＝6.0Hz，1H)，4.31-4.22(m，2H)，4.16(d，J＝9.0Hz，1H)，4.09(dd，J＝5.7，10.5Hz，1H)，3.94(d，J＝3.0Hz，1H)，3.85(dd，J＝6.3，11.1Hz，1H)，3.63(dd，J＝3.0，10.5Hz，1H)，2.95-2.85(m，1H)，2.93(s，3H)，2.45-2.13(m，3H)，2.13(s，3H)，1.84-1.03(m，13H)，0.87(t，J＝7.2Hz，3H)；MS(ESPOS)：431.3[M+H]；MS(ESNEG)：465.2[M+Cl].

Example 67

5-propyl-azepane-2-carboxylic acid [ cyclopentyl- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Lincosamide 23g (R) as described in general procedure Z ²⁰+R²¹Is cyclopentyl, R¹SMe) with 5-propyl-azaCoupling of-1, 2-dicarboxylic acid-1-tert-butyl ester gives the title compound. Reaction of intermediate 13a (R)¹＝SMe，R²⁰+R²¹Is cyclopentyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-3) under acidic conditions to give the title compound.

¹H NMR(300MHz，D₂O)δ5.33(d，J＝5.7Hz，1H)，4.24(dd，J＝4.8，9.0Hz，1H)，4.19-4.11(m，2H)，4.07(dd，J＝5.7，10.5Hz，1H)，3.90(d，J＝3.0Hz，1H)，3.62(dd，J＝3.3，10.5Hz，1H)，3.46(dd，J＝4.2，13.8Hz，1H)，3.19-3.08(m，1H)，2.36-2.09(m，3H)，2.12(s，3H)，2.08-1.81(m，2H)，1.80-1.40(m，8H)，1.36-1.01(m，7H)，0.83(t，J＝6.6Hz，3H)；MS(ESPOS)：445.2[M+H]；MS(ESNEG)：479.0[M+Cl].

Example 68

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -but-3-enyl ] -amide

Preparation of lincosamide 23f (R) as illustrated in scheme 23²⁰Vinyl radical, R²¹＝H，R¹＝SMe)。

¹H NMR(300MHz，CDCl₃)δ6.31(br d，J＝9.3Hz，1H)，5.80-5.62(m，1H)，5.64(d，J＝5.4Hz，1H)，5.41(dd，J＝0.9，3.0Hz，1H)，5.26(dd，J＝5.4，10.8Hz，1H)，5.23-5.10(m，3H)，4.42-4.25(m，2H)，3.56-2.44(m，1H)，2.35-2.23(m，1H)，2.14(s，3H)，2.09(s，3H)，2.08(s，3H)，1.97(m，3H)；MS(ESNEG)：470.0[M-H].

Preparation of lincosamide 23g (R) as described in scheme 23²⁰Vinyl radical, R²¹＝H，R¹＝SMe)。

¹H NMR(300MHz，CD₃OD)δ5.94-5.78(m，1H)，5.27(d，J＝5.7Hz，1H)，5.20-5.10(m，2H)，4.09(dd，J＝5.7，10.2Hz，1H)，4.04(dd，J＝1.5，3.3Hz，1H)，3.82(dd，J＝0.9，8.1Hz，1H)，3.57(dd，J＝3.3，9.9Hz，1H)，3.13(dt，J＝3.9，8.4Hz，1H)，2.57-2.47(m，1H)，2.14-2.02(m，1H)，2.07(s，3H)；MS(ESPOS)：272.0[M+Na]；MS(ESNEG)：248.2[M-H].

Lincosamide 23g (R) as described in general procedure Z²⁰Vinyl radical, R²¹＝H，R¹SMe) was coupled with 4-n-propyl gulonic acid to give the title compound.

¹HNMR(300MHz，D₂O)δ5.84-5.68(m，1H)，5.37(d，J＝5.7Hz，1H)，5.16-5.07(m，2H)，4.28-4.18(m，2H)，4.14-4.07(m，2H)，3.93(d，J＝3.3Hz，1H)，3.82(dd，J＝6.3，11.1Hz，1H)，3.66(dd，J＝3.3，10.5Hz，1H)，2.91-2.83(m，1H)，2.91(s，3H)，2.67-2.58(m，1H)，2.40-2.10(m，4H)，2.11(s，3H)，1.52-1.22(m，4H)，0.87(t，J＝72Hz，3H)；MS(ESPOS)：403.3[M+H]；MS(ESNEG)：437.0[M+Cl].

Example 69

4-propyl-piperidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -but-3-enyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰Vinyl radical, R²¹＝H，R¹SMe) with 4-propyl-piperidine-1, 2-dicarboxylic acid-1-tert-butyl ester 27b (R)⁹Propyl) to yield the title compound. Reaction of intermediate 13a (R)²⁰Vinyl radical, R²¹＝H，R¹＝SMe，R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

¹HNMR(300MHz，D₂O)δ5.82-5.66(m，1H)，5.37(d，J＝5.7Hz，1H)，5.16-5.07(m，2H)，4.24-4.05(m，3H)，3.92(d，J＝3.3Hz，1H)，3.85(dd，J＝3.3，12.9Hz，1H)，3.66(dd，J＝3.3，10.5Hz，1H)，3.47(brd，J＝12.3Hz，1H)，3.08-2.96(m，1H)，2.66-2.56(m，1H)，2.22-2.10(m，2H)，2.11(s，3H)，1.99-1.89(m，1H)，1.80-1.64(m，1H)，1.41-1.22(m，6H)，0.87(t，J＝6.6Hz，3H)；MS(ESPOS)：403.3[M+H]；MS(ESNEG)：437.2[M+Cl].

Example 70

5-propyl-azepane-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -butyl ] -amide

The title compound was prepared by coupling the cyclic amino acid 22f (R) prepared by general procedure T as described in general procedure Z⁹Is propyl radical, R^9bH, m-2) and lincosamine 23g (R)²⁰Ethyl, R²¹＝H，R¹SMe). Hydrogenation of the unsaturated intermediate affords the title compound.

¹H NMR(300MHz，D₂O)δ5.34(d，J＝5.7Hz，1H)，4.16-4.04(m，3H)，4.01(d，J＝9.3Hz，1H)，3.89(d，J＝3.3Hz，1H)，3.64(dd，J＝3.3，10.5Hz，1H)，3.45(dd，J＝5.1，13.2Hz，1H)，3.13(t，J＝12.0Hz，1H)，2.20-1.16(m，15H)，2.08(s，3H)，0.86(t，J＝7.5Hz，3H)，0.83(t，J＝6.9Hz，3H)；MS(ESPOS)：419.0[M+H]；MS(ESNEG)：453.2[M +Cl].

Example 71

4-propyl-piperidine-2-carboxylic acid [1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -butyl ] -amide

Example 69 hydrogenation of the title compound afforded the title compound.

¹H NMR(300MHz，D₂O)δ5.34(d，J＝6.0Hz，1H)，4.14-4.04(m，2H)，4.00(d，J＝9.3Hz，1H)，3.91-3.83(m，2H)，3.64(dd，J＝3.3，10.5Hz，1H)，3.51-3.43(m，1H)，3.08-2.96(m，1H)，2.22-2.13(m，1H)，2.07(s，3H)，1.99-1.89(m，1H)，1.83-1.65(m，2H)，1.48-1.13(m，9H)，0.85(t，J＝7.5Hz，6H)；MS(ESPOS)：405.4[M+H]；MS(ESNEG)：439.2[M+Cl].

Example 72

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ l- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -butyl ] -amide

Example 68 hydrogenation of the title compound afforded the title compound.

¹H NMR(300MHz，D₂O)δ5.36(d，J＝6.0Hz，1H)，4.28-4.03(m，4H)，3.93(d，J＝3.0Hz，1H)，3.85(dd，J＝6.9，11.1Hz，1H)，3.66(dd，J＝3.0，10.2Hz，1H)，2.95-2.85(m，1H)，2.93(s，3H)，2.47-2.19(m，3H)，2.10(s，3H)，1.86-1.70(m，1H)，1.54-1.16(m，7H)，0.87(t，J＝6.9Hz，6H)；MS(ESPOS)：405.4[M+H]；MS(ESNEG)：439.2[M+Cl].

Example 73

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ (4-chloro-phenyl) - (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Preparation of lincosamide 23f (R) as illustrated in scheme 23²⁰+R²¹4-chlorophenyl, R¹＝SMe)。

¹H NMR(300MHz，CDCl₃)□7.34(d，J＝8.4Hz，2H)，7.25(d，J＝8.4Hz，2H)，7.16(brd，J＝9.0Hz，1H)，5.55(d，J＝5.4Hz，1H)，5.50(d，J＝2.1Hz，1H)，5.30-5.13(m，3H)，4.66(d，J＝8.7Hz，1H)，2.09(s，3H)，2.07(s，3H)，1.98(s，3H)，1.65(s，3H)；MS(ESPOS)：563.9[M+Na]；MS(ESNEG)：539.8[M-H].

Preparation of lincosamide 23g (R) as described in scheme 23²⁰+R²¹4-chlorophenyl, R¹＝SMe)。

¹H NMR(300MHz，CD₃OD)□7.37(d，J＝8.4Hz，2H)，7.31(d，J＝9.0Hz，2H)，5.09(d，J＝6.0Hz，1H)，4.13-4.03(m，4H)，3.58(dd，J＝3.3，10.2Hz，1H)，1.41(s，3H)；MS(ESPOS)：320.0[M+H]；MS(ESNEG)：354.0[M+Cl].

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹4-chlorophenyl, R¹SMe) was coupled with 4-n-propyl gulonic acid to give the title compound.

¹H NMR(300MHz，D₂O)□7.43(d，J＝8.1Hz，2H)，7.36(d，J＝8.4Hz，2H)，5.18(d，J＝6.0Hz，1H)，5.12(d，J＝10.2Hz，1H)，4.48(d，J＝9.9Hz，1H)，4.29(dd，J＝5.4，9.0Hz，1H)，4.14-4.05(m，2H)，3.78(dd，J＝5.7，10.8Hz，1H)，3.70(dd，J＝3.3，10.2Hz，1H)，2.92(s，3H)，2.87(t，J＝10.8Hz，1H)，2.26-2.11(m，2H)，2.07-1.94(m，1H)，1.52(s，3H)，1.46-1.12(m，4H)，0.81(t，J＝7.2Hz，3H)；MS(ESPOS)：473.2[M+H]；MS(ESNEG)：507.2[M+Cl].

Example 74

4-propyl-piperidine-2-carboxylic acid [ (4-chloro-phenyl) - (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰+R²¹4-chlorophenyl, R¹SMe) and 27b (R)⁹Propyl) to yield the title compound.

¹H NMR(300MHz，CD₃OD)δ7.35(s，4H)，5.20(d，J＝9.6Hz，1H)，5.14(d，J＝6.0Hz，1H)，4.36(d，J＝9.3Hz，1H)，4.10(dd，J＝5.7，10.2Hz，1H)，3.99(d，J＝3.0Hz，1H)，3.89(dd，J＝3.0，12.6Hz，1H)，3.59(dd，J＝3.3，10.2Hz，1H)，3.45-3.36(m，1H)，3.04(dt，J＝3.3，13.2Hz，1H)，2.24-2.14(m，1H)，1.98-1.88(m，1H)，1.81-1.66(m，1H)，1.52(s，3H)，1.46-1.13(m，6H)，0.94(t，J＝7.2Hz，3H)；MS(ESPOS)：473.2[M+H]；MS(ESNEG)：507.2[M+Cl].

Example 75

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-isopropylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Preparation of lincosamide 23f (R) as illustrated in scheme 23²⁰Methyl, R²¹Methyl, R¹Isopropylthio).

¹H NMR(300MHz，CDCl₃)δ6.10(br d，J＝10.5Hz，1H)，5.79(d，J＝5.4Hz，1H)，5.36(dd，J＝1.2，3.3Hz，1H)，5.20(dd，J＝5.7，11.1Hz，1H)，5.09(dd，J＝3.3，10.8Hz，1H)，4.36(dd，J＝0.9，9.9Hz，1H)，4.26(dt，J＝3.0，10.2Hz，1H)，3.04-2.90(m，1H)，2.13(s，3H)，2.07(s，3H)，1.97(s，3H)，1.29(d，J＝4.8Hz，3H)，1.27(d，J＝5.1Hz，3H)，0.91(d，J＝6.9Hz，3H)，0.86(d，J＝6.9Hz，3H)

MS(ESPOS)：524.0[M+Na]

MS(ESNEG)：500.0[M-H]

Preparation of lincosamide 23g (R) as described in scheme 23²⁰Methyl, R²¹Methyl, R¹Isopropylthio).

¹H NMR(300MHz，CD₃OD)δ5.36(d，J＝6.0Hz，1H)，4.05(dd，J＝5.7，10.2Hz，1H)，4.01(dd，J＝1.5，3.3Hz，1H)，3.95(dd，J＝1.2，8.7Hz，1H)，3.48(dd，J＝3.3，10.5Hz，1H)，3.04-2.93(m，1H)，2.89(dd，J＝3.6，8.4Hz，1H)，2.07-1.95(m，1H)，1.30(d，J＝6.9Hz，3H)，1.26(d，J＝6.9Hz，3H)，0.98(d，J＝6.9Hz，3H)，0.87(d，J＝6.6Hz，3H)；MS(ESPOS)：280.0[M+H]；MS(ESNEG)：278.2[M-H]

Lincosamide 23g (R) as described in general procedure Z²⁰Methyl, R²¹Methyl, R¹Isopropylthio) was coupled with 4-n-propyl gulonic acid to give the title compound.

¹H NMR(300MHz，D₂O)δ5.47(d，J＝6.0Hz，1H)，4.25(brt，J＝7.2Hz，1H)，4.16(br s，2H)，4.07(dd，J＝5.7，10.5Hz，1H)，3.83(dd，J＝8.1，11.4Hz，2H)，3.56(dd，J＝3.0，10.5Hz，1H)，3.11-2.99(m，1H)，2.91(s，3H)，2.88(brt，J＝11.1Hz，1H)，2.45-2.20(m，3H)，2.15-2.00(m，1H)，1.50-1.37(m，2H)，1.36-1.23(m，8H)，0.90-0.80(m，9H)；MS(ESPOS)：433.4[M+H]；MS(ESNEG)：467.2[M+Cl]

Example 76

4-propyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-isopropylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰Methyl, R²¹Methyl, R¹Isopropylthio) and 4-propyl-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 27b (R)⁹Propyl) to yield the title compound. Reaction of intermediate 13a (R)¹Is isopropylthio, R ²⁰Methyl, R²¹Methyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

¹H NMR(300MHz，D₂O)δ5.47(d，J＝6.0Hz，1H)，4.14(brs，2H)，4.07(dd，J＝6.0，10.5Hz，1H)，3.89(dd，J＝3.0，12.6Hz，1H)，3.83(d，J＝3.3Hz，1H)，3.56(dd，J＝3.3，10.5Hz，1H)，3.47(br d，J＝13.5Hz，1H)，3.12-2.96(m，2H)，2.25-1.90(m，3H)，1.80-1.66(m，1H)，1.50-1.22(m，12H)，0.90-0.79(m，9H)；MS(ESPOS)：433.4[M+H]；MS(ESNEG)：467.2[M+Cl].

Example 77

1-methyl-4-propyl-pyrrolidine-2-carboxylic acid [1- (6-tert-butylsulfanyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide

Preparation of lincosamide 23f (R) as illustrated in scheme 23²⁰Methyl, R²¹Methyl, R¹Tert-butylthio).

¹H NMR(300MHz CDCl₃)δ6.08(br d，J＝9.3Hz，1H)，5.81(d，J＝5.7Hz，1H)，5.34(d，J＝3.0Hz，1H)，5.19(dd，J＝5.7，11.1Hz，1H)，5.01(dd，J＝3.3，11.1Hz，1H)，4.34-4.18(m，2H)，2.20-2.05(m，1H)，2.13(s，3H)，2.07(s，3H)，1.97(s，3H)，1.32(m，9H)，0.89(t，J＝6.6Hz，6H)；MS(ESPOS)：538.0[M+Na]；MS(ESNEG)：514.2[M-H].

Preparation of lincosamide 23g (R) as described in scheme 23²⁰Methyl, R²¹Methyl, R¹Tert-butylthio).

¹H NMR(300MHz，CD₃OD)δ5.39(d，J＝5.7Hz，1H)，4.05(dd，J＝6.0，10.8Hz)，1H)，4.01(dd，J＝1.2，3.3Hz，1H)，3.90(dd，J＝1.5，8.7Hz，1H)，3.39(dd，J＝3.3，10.5Hz，1H)，2.88(dd，J＝3.6，8.1Hz，1H)，2.08-1.95(m，1H)，1.36(s，9H)，0.98(d，J＝6.9Hz，3H)，0.89(d，J＝6.9Hz，3H)；MS(ESPOS)：294.0[M+H]；MS(ESNEG)：292.2[M-H]

Lincosamide 23g (R) as described in general procedure Z²Is isopropyl, R¹Tert-butylthio) was coupled with 4-n-propyl gulonic acid to give the title compound.

¹H NMR(300MHz，D₂O)δ5.50(d，J＝5.7Hz，1H)，4.26(br t，J＝7.5Hz，1H)，4.15(br s，2H)，4.08(dd，J＝5.7，10.5Hz，1H)，3.90-3.82(m，2H)，3.50(dd，J＝3.0，10.8Hz，1H)，2.93(s，3H)，2.91(brt，J＝11.1Hz，1H)，2.48-2.25(m，3H)，2.16-2.04(m，1H)，1.52-1.26(m，4H)，1.37(m，9H)，0.88(t，J＝6.9Hz，9H)；MS(ESPOS)：447.4[M+H]；MS(ESNEG)：481.2[M+Cl].

Example 78

4-propyl-piperidine-2-carboxylic acid [1- (6-tert-butylsulfanyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide

Lincosamide 23g (R) as described in general procedure Z²⁰Methyl, R²¹Methyl, R¹Tert-butylthio) and 4-propyl-piperidine-1, 2-dicarboxylic acid-1-tert-butyl ester 27b (R)⁹Propyl) to yield the title compound. Reaction of intermediate 13a (R)¹Tert-butylthio, R²⁰Methyl, R²¹Methyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

¹H NMR(300MHz，D₂O)δ5.50(d，J＝5.7Hz，1H)，4.13(brs，2H)，4.08(dd，J＝5.7，10.5Hz，1H)，3.92(dd，J＝3.0，12.6Hz，1H)，3.85(d，J＝3.3Hz，1H)，3.51(dd，J＝3.0，10.5Hz，2H)，3.06(brt，J＝10.8Hz，1H)，2.28-2.19(m，1H)，2.16-1.93(m，2H)，1.85-1.69(m，1H)，1.54-1.27(m，6H)，1.38(s，9H)，0.94-0.83(m，9H)；MS(ESPOS)：447.4[M+H]；MS(ESNEG)：481.0[M+Cl].

Example 79

4- (2-cyclopropyl-ethyl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

To a stirred suspension of 11b (0.5g, 1.9mmol, 1equiv), triphenylphosphine (39.9mg, 0.15mmol, 0.08equiv), copper (I) iodide (28.9mg, 0.15mmol, 0.08equiv), palladium acetate (17mg, 0.076mmol, 0.04equiv) in triethylamine (7mL) was added cyclopropylacetylene (Aldrich) (0.25g, 3.8mmol, 2equiv) under dry nitrogen. The mixture was stirred at rt overnight. The solvent was removed under vacuum to give a dark residue. The residue was purified by column chromatography to give 11c (R)⁹' -2-cyclopropyl-eth-1-ynyl) (0.39g, 100%) as a yellow oil.

¹H NMR(300MHz，CDCl₃)δ8.65-8.56(m，1)，8.06-7.99(m，1)，7.40-7.32(m，1)，3.98(s，3)，1.50-1.40(m，1)，0.96-0.81(m，4)；MS(ESPOS)：202.0[M+H]⁺.

To 11c (R)⁹' -2-cyclopropyl-eth-1-ynyl) (0.39g, 1.9mmol) in methanol (15mL) was added 10% palladium on carbon (0.2 g). The mixture was purged with hydrogen and aerated (1atm) and stirred at rt overnight. The palladium was removed by filtration, and the filtrate was concentrated to give 4- (2-cyclopropylethyl) -pyridine-2-carboxylic acid methyl ester (0).38g, 97%) as yellow oil (intermediate not shown).

¹H NMR(300MHz，CDCl₃)δ8.60(d，J＝4.5，1)，8.00-7.96(m，1)，7.34-7.29(m，1)，3.99(s，3)，2.78(t，J＝7.6，2)，1.58-1.49(m，2)，0.71-0.59(m，1)，0.47-0.38(m，2)，0.06-0.02(m，2)；MS(ESPOS)：228.2[M+Na]⁺

To a mixture of 4- (2-cyclopropylethyl) -pyridine-2-carboxylic acid methyl ester (0.38g) in MeOH (8mL) and water (8mL) was added concentrated HCl (158. mu.L) and platinum oxide (0.2 g). The mixture was purged with hydrogen and aerated (1atm), and stirred overnight. Platinum oxide was removed by filtration and the filtrate was evaporated to give a pale yellow solid 11d (R) ⁹2-cyclopropylethyl) was used without further purification.

To the above crude residue 11d (R)⁹2-cyclopropylethyl) 2N NaOH (3.8mL) and tert-butanol (2mL) were added. The reaction mixture was stirred at rt for 2h, then di-tert-butyl dicarbonate (0.62g, 2.85mmol) was added and the mixture was stirred overnight. The solvent was removed under vacuum and the resulting residue was diluted with water and then washed with ether. The aqueous layer was acidified with 2N HCl to pH 2.0 and extracted twice with ethyl acetate. The organic layers were combined and MgSO₄Drying and concentration gave 4- (2-cyclopropylethyl) -piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 11f (P ═ Boc, R)⁹2-cyclopropylethyl) (0.42g, 77%) as a clear syrup.

MS(ESPOS)：320.3[M+Na]⁺；MS(ESNEG)：296.2[M-H]^-.

Lincosamide 6b (R) as described in general procedure Z²＝H，R³Cl) with 4- (2-cyclopropylethyl) -piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester 11f (P ═ Boc, R⁹2-cyclopropylethyl) to yield the title compound. Reaction of intermediate 13a (R)²⁰Methyl, R²¹Methyl, R⁹Is propyl radical, P¹＝H，P²Tert-butyl-carboxylate, m-2) deprotected under acidic conditions to give the title compound.

MS(ESPOS)：451.3[M+H]⁺.

Example 80

4-Cyclopropylmethyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Preparation of 4-Cyclopropylmethylpyridine-2-carboxylic acid, Compound 10b (R), by general method O⁹Arbitrary ═ cyclopropylmethyl), the starting 4-cyclopropylmethylpyridine used was prepared by alkylating 4-methylpyridine with cyclopropylbromide by the method described by Osuch et al, Journal of the American Chemical Society, 1955, 78, 1723. The improved process is shown below.

To a solution of 4-methylpyridine (1.1g, 11.8mmol) in THF (5mL) at-78 deg.C was added LDA 2M in THF/heptane/ethylbenzene (Aldrich) (5.9mL, 11.8mmol), and the resulting reaction mixture was stirred at-78 deg.C for 3h, then at-40 deg.C for 1 h. Cyclopropyl bromide (1.43g, 11.8mmol) was then added at-78 deg.C, allowed to warm to room temperature, and stirred at room temperature for 1 h. Adding saturated aqueous NH to the reaction mixture₄Cl (10mL), aqueous phase extracted with EtOAc (10X 2mL), combined organic extracts over Na₂SO₄And (5) drying. The solvent was removed to give the product 4-cyclopropylmethylpyridine (0.5g, 31%) which was used without further purification.

Making lincosamide 6b (R) as in general method AA²＝H，R³═ Cl) with 4-cyclopropylmethylpyridine-2-carboxylic acid 10b (R)⁹(ii) cyclopropylmethyl) to afford intermediate 13b (R)¹＝SMe，R²＝Me，R³＝H，R⁹Is cyclobutyl-ethyl, P ¹H) to the title compound by catalytic hydrogenation.

MS(ESPOS)：437.2[M]⁺.

Example 81

4- (2-cyclobutyl-ethyl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Preparation of 4- (cyclobutyl-ethyl) -pyridine-2-carboxylic acid, Compound 10b (R), by general method O⁹Pivaloyl-ethyl), the starting material 4- (cyclobutyl-ethyl) -pyridine was prepared by alkylation of 4-methylpyridine with bromomethylcyclobutane as described in example 80.

Making lincosamide 6b (R) as in general method AA²＝H，R³═ Cl) and 4- (cyclobutyl-ethyl) -pyridine-2-carboxylic acid 10b (R)⁹Coupling to yield intermediate 13b (R)¹＝SMe，R²＝Me，R³＝H，R⁹Is cyclobutyl-ethyl, P¹H) to the title compound by catalytic hydrogenation.

MS(ESPOS)：465.2[M]⁺.

Example 82

4-cyclobutylmethyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl-1-amide

Preparation of 4-cyclobutylmethylpyridine-2-carboxylic acid, Compound 10b (R), by general method O⁹Cyclobutylmethyl), the starting material 4-cyclobutylpicoline used was prepared by alkylating 4-picoline with cyclobutylbromide as described in example 80.

Making lincosamide 6b (R) as in general method AA ²＝H，R³═ Cl) and 4-cyclobutylmethylpyridine-2-carboxylic acid, compound 10b (R)⁹4-cyclobutylmethyl) to afford intermediate 13b (R)¹＝SMe，R²＝Me，R³＝H，R⁹Is cyclobutylethyl, P¹H) to the title compound by catalytic hydrogenation.

MS (ESPOS)：451.2[M+H]⁺.

Example 83

4-Cyclopropylmethyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The protected amino acid intermediate, (2S, 4R) -4-cyclopropylmethyl-pyrrolidine-1, 2-dicarboxylic acid-1-tert-butyl ester, was prepared by means of the synthetic sequence described in Goodman et al, journal of organic Chemistry, 2003, 68, 3923, using cyclopropylmethyltriphenylphosphonium bromide (Aldrich) as starting material in the Wittig olefination step.

Lincolamine 6b (R) as described in general coupling scheme 11²＝H，R³Coupling of ═ Cl) with (2S, 4R) -4-cyclopropylmethyl-pyrrolidine-1, 2-dicarboxylic acid-1-tert-butyl ester afforded intermediate 13a (R)¹＝SMe，R²＝Me，R⁹Is cyclopropylmethyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 1), deprotection under acidic conditions afforded the title compound.

MS(ESPOS)：423.2[M+H]⁺.

Example 84

4- (2-Cyclobutylidene-ethyl) -pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The amino acid intermediate (2S, 4R) -4- (2-cyclobutylidene-ethyl) -pyrrolidine-1, 2-dicarboxylic acid-1-tert-butyl ester was prepared by alkylation of pyroglutamate 7a with (2-bromo-ethylene) -cyclobutane by means of general method K. The allylic halide (2-bromo-ethylene) -cyclobutane starting material is prepared in two steps from the cyclobutane ketone as described in U.S. patent No.3,711,555.

Enabling lincosamine 2b (R)¹＝SMe，R²Me) with protected amino acid 8c (R)⁹' -2-cyclobutylidene-ethyl) to give intermediate carbamate 13a (R)¹＝SMe，R²＝Me，R⁹2-cyclobutylidene-ethyl, P¹＝H，P²Boc, m 1) under acidic conditions to afford the title compound.

MS(ESPOS)：429.1[M+H]⁺.

Example 85

4- (2-Cyclobutylidene-ethyl) -pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Enabling lincosamine 6b (R)²＝H，R³Cl) with protected amino acid 8c (R)⁹' -2-cyclobutylidene-ethyl) to give intermediate carbamate 13a (R)¹＝SMe，R²＝Me，R⁹' 2-cyclobutylidene-ethyl, P¹＝H，P²Boc, m 1) under acidic conditions to afford the title compound.

MS(ESPOS)：450.1[M+H]⁺.

Example 86

4- (2-cyclobutyl-ethyl) -pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The amino acid intermediate (2S, 4R) -4- (2-cyclobutyl-ethyl) -pyrrolidine-1, 2-dicarboxylic acid-1-tert-butyl ester is prepared by alkylation of pyroglutamate 7a with (2-bromo-ethylene) -cyclobutane by means of general method K. The allylic halide (2-bromo-ethylene) -cyclobutane starting material is prepared in two steps from the cyclobutane ketone as described in U.S. patent No.3,711,555.

Enabling lincosamine 6b (R)²＝H，R³Cl) with protected amino acid 7d (R)⁹2-cyclobutyl-ethyl) coupling to give intermediate carbamate 13a (R)¹＝SMe，R²＝Me，R⁹Is cyclobutyl-ethyl, P¹＝H，P²Boc, m 1) under acidic conditions to afford the title compound.

MS(ESPOS)：451.2[M]⁺.

Example 87

4- (2-cyclopropyl-ethyl) -pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Enabling lincosamine 2b (R)¹＝SMe，R²Me) with protected amino acid 8c (R) prepared by general method M⁹2-cyclopropyl-ethyl) coupling to afford intermediate carbamate 13a (R)¹＝SMe，R²＝Me，R⁹Is cyclopropyl-ethyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 1), deprotection under acidic conditions afforded the title compound.

MS(ESPOS)：437.2[M+H]⁺.

Example 88

4-fluoro-1- (2-hydroxy-ethyl) -4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

As described in scheme 19, 4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide 18a (example 32) was treated with a solution of ethylene oxide in methanol to give the title compound.

MS(ESPOS)：473.3[M+H]⁺.

Example 89

4-butyl-4-fluoro-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Synthesis of Boc-protected 1-tert-butyl 4-fluoro-4-butyl-piperidine-1, 2-dicarboxylate 12d (P ═ Boc, m ═ 2, R) from the starting 1-tert-butyl (2S) -4-oxo-piperidine-1, 2-dicarboxylate⁹N-butyl) using the general procedure Q outlined in scheme 12, a 1-butyne anion was used as the four-carbon synthon in the 4-keto alkylation step. Preparation of starting 1-tert-butyl 4-oxo-piperidine-1, 2-dicarboxylate such as Bousquet, y.; anderson, p.c.; bogri, t.; DuceppeJ.; grenier, l.; guse, i.; tetrahedron, 1997, 53, 15671-15680.

Reacting lincosamide 6b (R) as described in general procedure Z²＝H，R³Cl) and 12d (P ═ Boc, m ═ 2, R⁹═ butyl) coupling to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹Is butyl, P¹＝H，P²Boc, m ═ 2), deprotection under acidic conditions afforded the title compound.

MS(ESPOS)：457.0[M+H]⁺.

Example 90

4-Cyclopropylmethyl-4-fluoro-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Reacting lincosamide 6b (R) as described in general procedure Z²＝H，R³C1) and 12d (P Boc, m 2, R)⁹(ii) cyclopropylmethyl) to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹Is cyclopropylmethyl, P¹＝H，P²Boc, m ═ 2), deprotection under acidic conditions afforded the title compound.

MS(ESPOS)：455.0[M+H]⁺.

Example 91

3-butyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

At 23 ℃ to carbamate 13a (R)²＝H，R³＝Cl，R⁹Is butyl, P¹＝H，P²Boc, m ═ 0) (82mg, 0.17mmol, 1equiv) in 1, 2-dichloroethane (10mL) was added H ₂O (0.40mL), followed by TFA (4.0 mL). After stirring at 23 ℃ for 20min, toluene (50mL) was added and the resulting solution was concentrated to dryness. The residue was purified by semi-preparative HPLC (Waters Nova-HR C18, 6 μm particle size,pore size, 20mmID X100 mm, H containing 5-60% acetonitrile₂O w/0.1% HCl, flow rate 20mL/min over 30 min) to give 41mg of the title compound as a white solid.

¹HNMR(300MHz，CD₃OD)δ5.30(d，J＝6.0Hz，1H)，4.64(d，J＝7.8Hz，1H)，4.63-4.52(m，2H)，4.29(d，J＝10.2Hz，1H)，4.07(dd，J＝5.7，10.2Hz，1H)，4.00(t，J＝6.6Hz，1H)，3.82(d，J＝3.3Hz，1H)，3.75(dd，J＝8.4，9.9Hz，1H)，3.56(dd，J＝3.3，10.2Hz，1H)，2.92-2.76(m，1H)，2.14(s，3H)，1.90-1.67(m，2H)，1.45(d，J＝6.6Hz，3H)，1.44-1.24(m，4H)，0.93(t，J＝6.9Hz，3H)；MS(ESPOS)：411.0[M+H]⁺.

Example 92

3-cyclopropylmethyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 25f (R)⁹(ii) cyclopropylmethyl) to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹Is cyclopropylmethyl, P¹＝H，P²Boc, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝5.7Hz，1H)，4.70(d，J＝7.5Hz，1H)，4.63-4.54(m，2H)，4.29(d，J＝9.9Hz，1H)，4.08(dd，J＝5.7，10.2Hz，1H)，4.02(t，J＝9.3Hz，1H)，3.88-3.80(m，2H)，3.57(dd，J＝3.3，10.2Hz，1H)，3.05-2.91(m，1H)，2.14(s，3H)，1.90-1.65(m，1H)，1.57-1.46(m，1H)，1.47(d，J＝6.6Hz，3H)，0.80-0.64(m，1H)，0.58-0.47(m，2H)，0.16-0.10(m，2H)；MS(ESPOS)：409.2[M+H]⁺.

Example 93

3-propyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 25f (R)⁹Propyl) to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹Is propyl radical, P¹＝H，P²Boc, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.31(d，J＝5.7Hz，1H)，4.66(d，J＝7.5Hz，1H)，4.63-4.54(m，2H)，4.31(d，J＝9.9Hz，1H)，4.09(dd，J＝5.4，10.2Hz，1H)，4.03(t，J＝9.6Hz，1H)，3.83-3.74(m，2H)，3.57(dd，J＝3.3，10.2Hz，1H)，2.95-2.80(m，1H)，2.15(s，3H)，1.88-1.66(m，2H)，1.47(d，J＝6.9Hz，3H)，1.46-1.30(m，2H)，0.97(t，J＝7.2Hz，3H)；MS(ESPOS)：397.0[M+H]⁺.

Example 94

3-butyl-1- (2-hydroxy-ethyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

3-butyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy) -prepared in example 91 was synthesized as described in scheme 19-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]Alkylation of amide samples with ethylene oxide (R)⁶2-hydroxyethyl) to yield the title compound.

¹H NMR(300MHz，CD₃OD)δ5.30(d，J＝5.4Hz，1H)，4.67-4.57(m，1H)，4.46-4.37(m，1H)，4.29-4.24(m，1H)，4.13-4.06(m，1H)，3.83-3.78(m，1H)，3.67-3.55(m，3H)，3.44-3.32(m，1H)，2.75-2.57(m，2H)，2.44-2.34(m，1H)，2.14(s，3H)，1.80-1.40(m，6H)，1.39-1.20(m，5H)，0.95-0.86(m，3H)；MS(ESPOS)：455.0[M+H]⁺.

Example 95

3-butyl-1-methyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

To Boc-carbamate 25f (R) at 23 deg.C⁹Butyl) (236mg, 0.92mmol, 1equiv) in aqueous formaldehyde (37%, 2.0mL) formic acid (95%, 1.0mL) was added. The resulting mixture was heated at reflux for 4h, then cooled to 23 ℃, treated with t-BuOH (5.0mL), and concentrated. Dissolving/suspending the crude residue in H₂O (15mL), frozen and lyophilized. The resulting solid was dissolved in 1.0N HCl (15mL), filtered, and concentrated. Dissolving/suspending the resulting product in H₂O, a cloudy suspension was obtained, filtered through a nylon membrane (0.2 μm), and concentrated to give 186mg of a white solid, the major component of which was the desired hydrochloride salt of 3-butyl-1-methyl-azetidine-2-carboxylic acid. The product was used without further purification.

MS(ESPOS)：172.3[M+H].

Such as general procedure Z lincosamine 6b (R)²＝H，R³═ Cl) with 3-butyl-1-methyl-azetidine-2-carboxylate to give the title compound 。

¹H NMR(300MHz，D₂O)δ5.40(d，J＝5.7Hz，1H)，4.73(d，J＝7.8Hz，1H)，4.66-4.56(m，1H)，4.48dd，J＝1.2，9.9Hz，1H)，4.38-4.27(m，2H)，4.11(dd，J＝5.7，10.5Hz，1H)，3.88(d，J＝3.0Hz，1H)，3.81(t，J＝9.6Hz，1H)，3.67(dd，J＝3.3，10.5Hz，1H)，3.04-2.87(m，1H)，2.94(s，3H)，2.18(s，3H)，1.90-1.68(m，2H)，1.44(d，J＝6.9Hz，3H)，1.40-1.22(m，4H)，0.87(t，J＝6.9Hz，3H)；MS(ESPOS)：425.3[M+H]⁺.

Example 96

3-pentyl-azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 25f (R)⁹Pivaloyl) to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹Pentyl radical, P¹＝H，P²Tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

H NMR(300MHz，CD₃OD)δ5.30(d，J＝5.4Hz，1H)，4.63-4.53(m，3H)，4.30(d，J＝9.6Hz，1H)，4.08(dd，J＝5.7，10.2Hz，1H)，4.00(t，J＝9.6Hz，1H)，3.81(d，J＝2.4Hz，1H)，3.74(dd，J＝7.8，9.9Hz，1H)，3.57(dd，J＝3.3，10.2Hz，1H)，2.92-2.78(m，1H)，2.15(s，3H)，1.9-1.67(m，2H)，1.46(d，J＝6.9Hz，3H)，1.44-1.26(m，6H)，0.92(t，J＝7.5Hz，3H)；MS(RSPOS)：425.0[M+H]⁺.

Example 97

3- (3-methyl-butyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 25f (R)⁹Coupling to 3-methyl-butyl) gave intermediate 13a (R)²＝H，R³＝Cl，R⁹3-methyl-butyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.31(d，J＝5.4Hz，1H)，4.63-4.53(m，2H)，4.30(d，J＝10.5Hz，1H)，4.08(dd，J＝5.7，10.5Hz，1H)，4.00(t，J＝9.6Hz，1H)，3.81(d，J＝2.4Hz，1H)，3.74(dd，J＝8.1，9.9Hz，1H)，3.57(dd，J＝3.3，10.2Hz，1H)，2.88-2.75(m，1H)，2.15(s，3H)，1.90-1.67(m，2H)，1.63-1.50(m，1H)，1.46(d，J＝6.9Hz，3H)，1.39-1.10(m，3H)，0.94(d，J＝1.5Hz，3H)，0.92(d，J＝1.5Hz，3H)；MS(ESPOS)：425.0[M+H]⁺.

Example 98

3- (3-cyclopropyl-propyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 26f (R)⁹Coupling to 3-cyclopropyl-propyl) gave intermediate 13a (R)²＝H，R³＝Cl，R⁹The compound is a compound of the formula (I) butyl group,P¹＝H，P²tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹HNMR(300MHz，CD₃OD)δ5.29(d，J＝5.7Hz，1H)，4.64-4.51(m，3H)，4.29(d，J＝10.2Hz，1H)，4.07(dd，J＝5.7，10.2Hz，1H)，3.99(t，J＝9.9Hz，1H)，3.79(d，J＝3.3Hz，1H)，3.74(dd，J＝8.4，9.9Hz，1H)，3.55(dd，J＝3.3，10.2Hz，1H)，2.91-2.77(m，1H)，2.13(s，3H)，1.93-1.68(m，2H)，1.60-1.32(m，2H)，1.44(d，J＝6.9Hz，3H)，1.24(q，J＝10.2Hz，2H)，0.74-0.62(m，1H)，0.44-0.36(m，2H)，0.04--0.02(m，2H)；MS(ESPOS)：437.2[M+H]⁺.

Example 99

3- (3-cyclobutyl-propyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 26f (R)⁹Coupling to 3-cyclobutyl-propyl) gave intermediate 13a (R)²＝H，R³＝Cl，R⁹3-cyclobutyl-propyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.29(d，J＝5.7Hz，1H)，4.63-4.46(m，3H)，4.28(d，J＝10.2Hz，1H)，4.08(dd，J＝5.4，10.2Hz，1H)，3.89(t，J＝9.0Hz，1H)，3.79(d，J＝3.6Hz，1H)，3.65(dd，J＝8.1，9.6Hz，1H)，3.56dd，J＝3.0，10.2Hz，1H)，2.86-2.71(m，1H)，2.38-2.20(m，1H)，2.14(s，3H)，2.10-1.96(m，2H)，1.90-1.52(m，6H)，1.44(d，J＝6.9Hz，3H)，1.44-1.14(m，4H).

MS(ESPOS)：451.2[M+H]

Example 100

3- (2-cyclobutyl-ethyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 26f (R)⁹2-cyclobutyl-ethyl) coupling to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹2-cyclobutyl-ethyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.29(d，J＝5.7Hz，1H)，4.63-4.50(m，3H)，4.29(d，J＝10.2Hz，1H)，4.08(dd，J＝5.4，10.2Hz，1H)，3.95(t，J＝9.3Hz，1H)，3.79(d，J＝3.0Hz，1H)，3.69(dd，J＝8.4，9.9Hz，1H)，3.56(dd，J＝3.3，10.2Hz，1H)，2.87-2.72(m，1H)，2.38-2.20(m，1H)，2.14(s，3H)，2.13-2.00(m，2H)，1.94-1.55(m，6H)，1.54-1.34(m，2H)，1.45(d，J＝6.6Hz，3H)；MS(ESPOS)：437.2[M+H]⁺.

Example 101

3- (2-cyclopropyl-ethyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 26f (R)⁹2-cyclopropyl-ethyl) coupling to afford intermediate 13a (R)²＝H，R³＝Cl，R⁹2-cyclopropyl-ethyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.29(d，J＝5.7Hz，1H)，4.62-4.50(m，3H)，4.29(d，J＝10.2Hz，1H)，4.07(dd，J＝5.4，10.2Hz，1H)，3.99(t，J＝9.6Hz，1H)，3.82-3.71(m，2H)，3.56(dd，J＝3.3，10.2Hz，1H)，2.95-2.80(m，1H)，2.13(s，3H)，2.00-1.77(m，2H)，1.44(d，J＝6.9Hz，3H)，1.35-1.20(m，2H)，0.74-0.62(m，1H)，0.48-0.40(m，2H)，0.09-0.02(m，2H)；MS(ESPOS)：423.2[M+H]⁺.

Example 102

3- (3, 3-difluoropropyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Such as general procedure Z lincosamine 6b (R)²＝H，R³Ci) and azetidinoic acid 26f (R)⁹Coupling to 3, 3-difluoropropyl) gave intermediate 13a (R)²＝H，R³＝Cl，R⁹2-cyclopropyl-ethyl, P¹＝H，P²Tert-butyl-carboxylate, m ═ 0), deprotection under acidic conditions afforded the title compound.

¹H NMR(300MHz，CD₃OD)δ5.93(t，J＝57Hz，1H)，5.29(d，J＝5.7Hz，1H)，4.64(d，J＝7.5Hz，1H)，4.60-4.51(m，2H)，4.29(d，J＝10.2Hz，1H)，4.07(dd，J＝5.7，10.2Hz，1H)，4.02(t，J＝8.7Hz，1H)，3.82-3.74(m，2H)，3.55(dd，J＝3.3，10.5Hz，1H)，2.96-2.82(m，1H)，2.13(s，3H)，2.06-1.76(m，4H)，1.44(d，J＝6.9Hz，3H)；MS(ESPOS)：433.0[M+H]⁺.

Example 103

4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide

2-methyl-1- (3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester.

To a solution of 2- (1-amino-2-methyl-propyl) -6-propyl-tetrahydro-pyran-3, 4, 5-triol (51.9mg, 0.21mmol, 1equiv) prepared by general method AB in anhydrous DMF (4.5mL) was added triethylamine (117 μ L, 0.84mmol, 4equiv) at 0 ℃, followed by BSTFA (84 μ L, 0.32mmol, 1.5 equiv). The reaction mixture was stirred at 0 ℃ for 10 min and then at RT for 45 min. To the reaction mixture is added the protected amino acid 7d (R) as prepared in general method L⁹N-pentyl, P ═ Boc) (72mg, 0.25mmol, 1.2equiv) and HATU (120mg, 0.32mmol, 1.5 equiv). The reaction mixture was stirred at RT for 2h, evaporated to dryness and dissolved in Et ₂O (150mL), with 10% citric acid (1X), saturated NaHCO₃(1x) and brine wash. The organic layer was purified over MgSO₄Dried and concentrated to give the crude product (190mg) as a yellow oil. The residue was dissolved in DCE (6mL), and trifluoroacetic acid (4mL) containing water (0.2mL) was added with stirring. The reaction mixture was stirred at RT for 1h, then the solvent was removed under vacuum by repeated co-evaporation from DCE. The residue was purified by silica column chromatography using 10% 0.25M NH₃Eluted with MeOH/DCM to afford the product (38.4mg, 43%).¹H NMR(300MHz，D₂O)δ4.37(dd，J＝4.7，9.1，1)，4.14(dd，J＝3.0，9.6，1)，3.97-3.90(m，2)，3.70(d，J＝2.8，1)，3.62-3.51(m，3)，2.89(dd，J＝9.1，11.3，1)，2.33-2.03(m，1)，1.64-1.55(m，2)，1.52-1.45(m，3)，1.36-1.25(m，6)，1.0-0.86(m，12)；MS (ESPOS)：416.6[M+H].

Example 104

4-propyl-piperidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Intermediate 10b (R) prepared in general procedure O was used⁹N-propyl) the title compound of example 2 was prepared according to the procedure described for example 103.

MS(ESPOS)：401.7[M+H]⁺.

Example 105

4-propyl-piperidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (2, 2, 2-trifluoro-ethylsulfanyl) -tetrahydro-pyran-2-yl ] -propyl } -amide

Intermediate 27a 4-propyl-piperidine-1, 2-dicarboxylic acid 1-tert-butyl ester prepared in general method.

MS(ESNEG)：270.2[M-H]^-.

The title compound was prepared according to general procedure R and scheme 16 using 1, 1, 1-trifluoroethanethiol as the thiol nucleophile. Coupling and deprotection of the protected amino acid intermediate, 1-tert-butyl 4-propyl-piperidine-1, 2-dicarboxylate, was performed as in example 103.

MS(ESPOS)：473.7[M+H]⁺.

Example 106

4-pentyl-pyrrolidine-2-carboxylic acid [1- (6-ethoxyethyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide

2- [1- (6-allyl-3, 4, 5-tris-benzyloxy-tetrahydro-pyran-2-yl) -2-methyl-propylcarbamoyl ] -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester.

To a stirred building block 15c (650mg, 1.26mmol, 1equiv) and a protected amino acid 7d (R) at 0 deg.C⁹Pentyl, P ═ Boc) (395mg, 1.39mmol, 1.1equiv) in anhydrous DMF (5.0mL) was added DIEA (0.88mL, 5.0mmol, 4equiv), followed by solid HATU (956mg, 2.52mmol, 2.0 equiv). The reaction mixture was stirred at RT for 3h, evaporated to dryness, dissolved in ethyl acetate and washed with 10% citric acid (1 ×), water (1 ×), saturated NaHCO₃(1x) and brine wash. Subjecting the organic layer to Na₂SO₄Drying and concentrating to obtain yellow syrup. The filtrate was concentrated and the residue was purified by column chromatography on silica eluting with 10% EtOAc/hexanes to 20% EtOAc/hexanes to give the product 2- [1- (6-allyl-3, 4, 5-tris-benzyloxy-tetrahydro-pyran-2-yl) -2-methyl-propylcarbamoyl]-4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester as a colorless oil (897mg, 89%).

2- { 2-methyl-1- [3, 4, 5-tris-benzyloxy-6- (2-hydroxy-ethyl) -tetrahydro-pyran-2-yl ] -propylcarbamoyl } -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester.

Stirring 2- [1- (6-allyl-3, 4, 5-tris-benzyloxy-tetrahydro-pyran-2-yl) -2-methyl-propylcarbamoyl at-78 ℃]-4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester (634mg, 0.81mmol, 1equiv) in DCM (60mL)A persistent pale blue color was observed after treatment with an ozone-containing oxygen stream for 20 min. After 30min, use N₂Excess ozone was removed by air flow, DMS (3mL) in DCM (10mL) was added and the solution was allowed to warm to RT overnight. The solution was evaporated to dryness, the residue was dissolved in EtOH (50mL), cooled to 0 ℃ and washed with NaBH₄(300mg, 8.1mmol, 10 equiv). After 1h, the excess NaBH is destroyed by acidifying the reaction mixture₄The solvent was removed and the crude product was purified by column chromatography on silica eluting with 20% EtOAc in hexanes to give the alcohol product (304mg, 47%).

2- { 2-methyl-1- [3, 4, 5-tris-benzyloxy-6- (2-ethoxy-ethyl) -tetrahydro-pyran-2-yl ] -propylcarbamoyl } -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester.

To a stirred solution of washed NaH (4.4mg, 0.183mmol, 1equiv) in THF (0.8mL) at 0 deg.C was added the alcohol intermediate 2- { 2-methyl-1- [3, 4, 5-tris-benzyloxy-6- (2-hydroxy-ethyl) -tetrahydro-pyran-2-yl- ] propylcarbamoyl } -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester (144mg, 0.183mmol, 1equiv), after 10min EtI (73. mu.L, 0.92mmol, 5.0equiv) was added and the reaction mixture was stirred overnight. The reaction mixture was evaporated to dryness. The filtrate was concentrated and the residue was purified by preparative chromatography eluting with 30% EtOAc/hexanes to give the product 2- { 2-methyl-1- [3, 4, 5-tris-benzyloxy-6- (2-ethoxy-ethyl) -tetrahydro-pyran-2-yl ] -propylcarbamoyl } -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester (33.8mg, 22%) as a colorless oil.

4-pentyl-pyrrolidine-2-carboxylic acid [1- (6-ethoxymethyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide.

2- { 2-methyl-1- [3, 4, 5-tri-benzyloxy-6- (2-ethoxy-ethyl) -tetrahydro-pyran-2-yl]-Propylcarbamoyl } -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester (33.8mg) and degussa 50% w/w wet 10% palladium on carbon (80mg) were suspended in MeOH (3mL) under 1atm pressure H₂Stirring for 20 h. The reaction mixture was filtered through celite, evaporated to dryness to give the crude product, which was purified by silica column chromatography eluting with 3% to 5% MeOH in DCM,the Boc-protected ether product (19mg) was obtained, dissolved in DCE (1mL), and trifluoroacetic acid (1mL) containing water (0.05mL) was added with stirring. The reaction mixture was stirred at RT for 1h, then the solvent was removed under vacuum by repeated co-evaporation from DCE. The TFA salt was lyophilized from 1: 1 MeCN/water with excess dilute HCl to give the title compound (13.0mg, 66%).

¹H NMR(300MHz，D₂O)δ4.47-4.38(m，1)，4.21-4.16(m，1)，4.11-4.06(m，1)，3.96(dd，J＝6.3，9.6，1)，3.81(s，1)，3.61-3.50(m，7)，2.92(dd，J＝9.9，9.9，1)，2.33-1.98(m，7)，1.96-1.82(m，1)，1.47-1.33(m，11)，1.18(t，J＝6.9，3)，0.97-0.89(m，12)；MS(ESPOS)：446.4[M+H].

Example 107

1- (2-hydroxy-ethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide

The title compound was prepared according to the procedure described in scheme 19. Ethylene oxide is used as the alkylating agent. To a stirred solution of 4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl at 0 deg.C ]Amide (example 103) (12.0mg 0.029mmol, 1equiv) in MeOH (2mL) with TEA (100. mu.L) condensed (condensed) ethylene oxide (200. mu.L) was added and the reaction mixture was stirred for 48 h. The reaction mixture was evaporated to dryness and the residue was purified by column chromatography on silica using 20% 0.25M NH₃Eluted with MeOH/DCM to give the crude N-alkylated product. The crude product was dissolved in Et₂O, filtration and the filtrate taken in Et 2M HCl₂Treatment with O solution, collection of the precipitated HCl salt, and Et₂O wash, freeze dry to give the title compound as a colorless powder (4.4mg, 34%).

MS(ESPOS)：473.6[M+H]⁺.

Example 108

4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -propyl ] -amide

2- { 2-methyl-1- [3, 4, 5-tri-hydroxy-tetrahydro-pyran-2-yl ] -propylcarbamoyl } -4-pentyl-pyrrolidine-1-carboxylic acid tert-butyl ester.

In N₂Next, to a stirred solution of moist Raney nickel R1(300mg) suspended in EtOH (5mL) was added 1- (2- (S) -4- (R) -N-pentylpyrrolidin-2-yl) -N- {1- (R) - [2- (S) -3- (S), 4- (S), 5- (R) -trihydroxy-6- (R) - (methylthio) tetrahydropyran-2-yl]-2-methylpropan-1-yl } acetamide (85.0mg, 0.164mmol, 1equiv) in EtOH (5 mL). The reaction mixture was refluxed for 2h, cooled to RT, filtered through celite, evaporated to dryness to give the crude product (66mg) which was purified by silica column chromatography eluting with 3% MeOH in DCM to give the N-Boc-protected desulphatomethyl product (42.7mg, 55%).

TLC Rf＝0.27(10％MeOH/DCM)；MS(ESPOS)：473.6[M+H]⁺(ESNEG)：507.5[M+HC l].

4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -propyl ] -amide.

The N-Boc-protected product, the devothiomethylated product, was dissolved in DCE (5mL) and trifluoroacetic acid (5mL) containing water (0.1mL) was added with stirring. The reaction mixture was stirred at RT for 40min, then the solvent was removed under vacuum by repeated co-evaporation from DCE. The residue was dissolved in 1: 1 MeCN/water cooled to 0 deg.C, 1M HCl (0.5mL) was added, the solution was filtered and lyophilized to give the title compound (26mg, 43%) as a colorless powder.

TLC(CHCl₃：MeOH：32％aq.AcOH)Rf＝0.58；MS(ESPOS)387.3[M+H]⁺.

Examples 109-127, 142 and 143 can be prepared as described herein.

Example 109

Example 110

Example 111

5-propyl-azepane-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-isopropylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 112

5-propyl-azepane-2-carboxylic acid [1- (6-tert-butylsulfanyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide

Example 113

5-propyl-azepane-2-carboxylic acid [ (4-chlorophenyl) - (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -methyl ] -amide

Example 114

Example 115

4-pentyl-pyrrolidine-2-carboxylic acid [1- (6-butoxy-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide

Example 116

4-butyl-1-methyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 117

Phosphoric acid mono- (4, 5-dihydroxy-6- { 2-methyl-1- [ (4-pentyl-pyrrolidine-2-carbonyl) -amino ] -propyl } -2-propyl-tetrahydro-pyran-3-yl) ester

Example 118

Hexadecanoic acid 4, 5-dihydroxy-6- { 2-methyl-1- [ (4-pentyl-pyrrolidine-2-carbonyl) -amino ] -propyl } -2-propyl-tetrahydro-pyran-3-yl ester

Example 119

Phosphoric acid mono- (4, 5-dihydroxy-6- { 2-methyl-1- [ (4-propyl-pyrrolidine-2-carbonyl) -amino ] -propyl } 2-propyl-tetrahydro-pyran-3-yl) ester

Example 120

Hexadecanoic acid 4, 5-dihydroxy-6- { 2-methyl-1- [ (4-propyl-pyrrolidine-2-carbonyl) -amino ] -propyl } -2-propyl-tetrahydro-pyran-3-yl ester

Example 121

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-pentyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 122

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propylcarbamoyl]-4-pentyl-pyrrolidine-1-carboxylic acid 5-methyl-2-oxo- [1, 3]Dioxolen-4-ylmethyl esters

Example 123

1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-propyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 124

2- [ 2-methyl-1- (3, 4, 5-trihydroxy-6-propyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-propyl-pyrrolidine-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester

Example 125

4-propyl-pyrrolidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (2-hydroxy-ethyl) -tetrahydro-pyran-2-yl ] -propyl } -amide

Example 126

4-propyl-pyrrolidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (3-hydroxy-propyl) -tetrahydro-pyran-2-yl ] -propyl } -amide

Example 127

4-propyl-pyrrolidine-2-carboxylic acid [ 2-methyl-1- (3, 4, 5-trihydroxy-6-hydroxymethyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 142

4-propyl-pyrrolidine-2-carboxylic acid { 2-methyl-1- [3, 4, 5-trihydroxy-6- (2-methylsulfanyl-ethyl) -tetrahydro-pyran-2-yl ] -propyl } -amide

Example 143

4-propyl-pyrrolidine-2-carboxylic acid [1- (6-cyclopropylmethyl-3, 4, 5-trihydroxy-tetrahydro-pyran-2-yl) -2-methyl-propyl ] -amide

The following specific prodrug examples 128-141 were prepared from the respective parent compounds (see above) using the methods described above.

Example 128

Phosphoric acid mono- (6- { 2-chloro-1- [ (5-propyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester

Example 129

Hexadecanoic acid 6- { 2-chloro-1- [ (5-propyl-azepane-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester

Example 130

Phosphoric acid mono- (6- { 2-chloro-1- [ (5-fluoro-5-propyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester

Example 131

Hexadecanoic acid 6- { 2-chloro-1- [ (5-fluoro-5-propyl-azepane-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester

Example 132

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -5-fluoro-5-propyl-azepan-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester

Example 133

5-fluoro-1- (5-methyl-2-oxo- [1, 3)]Dioxol-4-ylmethyl) -5-propyl-azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl]-amides of

Example 134

5-Cyclopropylmethyl-1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -azepane-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example 135

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -5-cyclopropylmethyl-azepan-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester

Example 136

Hexadecanoic acid 6- { 2-chloro-1- [ (5-cyclopropylmethyl-azepane-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester

Example 137

Phosphoric acid mono- (6- { 2-chloro-1- [ (5-cyclopropylmethyl-azepan-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester

Example 138

Hexadecanoic acid 6- { 2-chloro-1- [ (4-fluoro-4-propyl-piperidine-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl ester

Example 139

Phosphoric acid mono- (6- { 2-chloro-1- [ (4-fluoro-4-propyl-piperidine-2-carbonyl) -amino ] -propyl } -4, 5-dihydroxy-2-methylsulfanyl-tetrahydro-pyran-3-yl) ester

Example 140

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -4-fluoro-4-propyl-piperidine-1-carboxylic acid 5-methyl-2-oxo [1, 3] dioxol-4-ylmethyl ester

Example 141

4-fluoro-1- (5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl) -4-propyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide

Example A

Sensitivity test

Compounds were tested in compliance with the microdilution method of NCCLS (National Committee for clinical Laboratory standards. methods for differential biological reporting tests for bacterial that grow in bacteria, applied standard-Fifth evaluation. NCCLS recording M7-A5, NCCLS, Wayne, PA.2000; National Committee for clinical Laboratory standards. methods for analytical reporting biological stability of microbiological bacteria; applied standard-modification. NCCLS recording M11-A4, CLS, NCCLS recording, PA.2001). The assay was performed in a sterile plastic 96-well microtiter plate (Greiner) with round-bottom wells.

Preparation of Compounds

Stock solutions of test compound and control antibiotic were prepared in DMSO at a concentration of 10 mg/ml. Serial 2-fold dilutions of each drug were made across each row in microtiter plates, using DMSO as solvent, at 100-fold higher concentrations than the desired final concentration. Columns 1-11 wells contain drug and column 12 serves as a biological growth control, without drug. Each well in the master plate was diluted with sterile deionized water, mixed, and a volume of 10 μ L was dispensed to each well of the resulting assay plate.

Preparation of inoculum

Using Microbank^TMMethod (Pro-Lab Diagnostics) stock cultures were prepared and stored at-80 ℃. To propagate aerobic strains, one bead was removed from the frozen vial, aseptically streaked onto tryptic soy agar (Difco), chocolate agar (Remel), or blood agar (Remel), and incubated overnight at 35 ℃. Anaerobic bacteria were cultured in Brucella agar (Remel) supplemented with hemin and vitamin K and incubated anaerobically at 35 ℃ for 24 to 48h using anaerobic jars (Mitsubishi). Standardized inocula were prepared using the direct colony suspension method (National Committee for Clinical Laboratory standards. methods for differential antimicrobial activity tests for bacterial present microorganism; applied standard-replication.NCCLS administration M7-A5, NCCLS, Wayne, PA.2000; National Committee for Clinical Laboratory standards. methods for antimicrobial activity testing of anaerobic bacteria; applied standard-replication.NCCLS administration M11-A4, NCCLS, Wayne, PA.2001) according to NCCLS guidelines. Isolated colonies were selected from 18-24hr agar plates and resuspended in 0.9% sterile saline to match the 0.5McFarland turbidity standard. The suspension was used within 15 minutes of preparation.

Streptococcus pneumoniae VSPN1001	Streptococcus pneumoniae ATCC 49619
Streptococcus pneumoniae VSPN1001	Streptococcus pneumoniae ATCC 49619	Streptococcus pneumoniae VSPN3026	Streptococcus pneumoniae R6x

Streptococcus pneumoniae VSPN1001	Streptococcus pneumoniae ATCC 49619
Streptococcus pneumoniae VSPN1001	Streptococcus pneumoniae ATCC 49619	Streptococcus pneumoniae VSPN4054	Streptococcus pneumoniae 488K
Streptococcus pneumoniae VSPN4021	Streptococcus pneumoniae 9	Streptococcus pneumoniae VSPN4054	Streptococcus pneumoniae 488K
Streptococcus pneumoniae VSPN4021	Streptococcus pneumoniae 9	Staphylococcus aureus VSAU1017	Staphylococcus aureus Smith
Staphylococcus aureus VSAU1003	Staphylococcus aureus ATCC 25923	Staphylococcus aureus VSAU1017	Staphylococcus aureus Smith
Staphylococcus aureus VSAU1003	Staphylococcus aureus ATCC 25923	Staphylococcus aureus VSAU4020	Staphylococcus aureus 125
Staphylococcus aureus VSAU4048	Staphylococcus aureus 85-EPI	Staphylococcus aureus VSAU4020	Staphylococcus aureus 125
Staphylococcus aureus VSAU4048	Staphylococcus aureus 85-EPI	Staphylococcus aureus VSAU4065	Staphylococcus aureus VSAU4065
Staphylococcus epidermidis VSEP1001	Staphylococcus epidermidis ATCC 12228	Staphylococcus aureus VSAU4065	Staphylococcus aureus VSAU4065
Staphylococcus epidermidis VSEP1001	Staphylococcus epidermidis ATCC 12228	Enterococcus faecalis VEFL1003	Enterococcus faecalis ATCC 51299
Enterococcus faecium VEFA1005	Enterococcus faecium BM4147.1	Enterococcus faecalis VEFL1003	Enterococcus faecalis ATCC 51299
Enterococcus faecium VEFA1005	Enterococcus faecium BM4147.1	Haemophilus influenzae VHIN1003	Haemophilus influenzae ATCC 49766
Haemophilus influenzae VHIN1004	Haemophilus influenzae ATCC 31517	Haemophilus influenzae VHIN1003	Haemophilus influenzae ATCC 49766
Haemophilus influenzae VHIN1004	Haemophilus influenzae ATCC 31517	Haemophilus influenzae VHIN1005acr	Haemophilus influenzae LS-2
Moraxella catarrhalis VMCA1001	Moraxella catarrhalis ATCC 25238	Haemophilus influenzae VHIN1005acr	Haemophilus influenzae LS-2
Moraxella catarrhalis VMCA1001	Moraxella catarrhalis ATCC 25238	Escherichia coli VECO2096	Escherichia coli MG1655
Escherichia coli VECO2526 tolC	Escherichia coli MG1655 tolC	Escherichia coli VECO2096	Escherichia coli MG1655
Escherichia coli VECO2526 tolC	Escherichia coli MG1655 tolC	Bacteroides fragilis VBFR1001	Bacteroides fragilis ATCC 25285
Bacteroides thetaiotaomicron VBTH1001	Bacteroides thetaiotaomicron ATCC #29741	Bacteroides fragilis VBFR1001	Bacteroides fragilis ATCC 25285
Bacteroides thetaiotaomicron VBTH1001	Bacteroides thetaiotaomicron ATCC #29741	Clostridium difficile VCDI1001	Clostridium difficile ATCC 9689

Preparation of MIC assay plates

Media was prepared at 1.1x concentration: Mueller-Hinton broth MHB (Difco) recommended by NCCLS, supplemented with Ca + + and Mg + +; MHB, supplemented with 5% horse lysed blood; HTM broth (Remel); or Brucella broth (Remel) supplemented with hemin and vitamin K. For each organism, the standardized suspension is diluted into the appropriate growth medium in a sterile reservoir. After mixing, the drug-containing assay plate wells were seeded with a volume of 90 μ L. Thus, for each MIC assay, each well contained a final volume of 100. mu.L, and the inoculum size was approximately 5X 10⁵cfu/ml, DMSO does not exceed 1%.

Interpretation of MIC

The finished microtiter plates were incubated at 35 ℃ for 16-20h in ambient air for aerobic bacteria and at 35 ℃ for 46-48 h for anaerobic bacteria or in anaerobic tanks (Mitsubishi). The optical density of each well was determined at 600nm using a VersaMax Microplate reader (Molecular Devices, Sunnyvale, Calif.). MI C was defined as the lowest drug concentration that resulted in complete inhibition of visible bacterial growth.

Example 47 isomer I and example 47 isomer II possess in vitro potency against the gram-negative organism Haemophilus influenzae with a MIC ≦ 4 μm/mL. Example 47 isoforms 1 and 2 further showed a MIC of 0.5 μm/mL against Haemophilus influenzae strain ATCC 31517, in contrast to clindamycin which showed a MIC of 8 μm/mL against Haemophilus influenzae strain ATCC 31517.

Example B

Efficacy in murine Staphylococcus aureus septicemia

According to a model published in another place, efficacy studies were carried out in the Staphylococcus aureus murine septicemia model (Goldstein, B.P., G.Candiani, T.M.Arain, G.Romano, I.Ciciliato, M.Berti, M.Abbondi, R.Scotti, M.Maineii, F.Ripamonti et al, 1995. antimicrobial activity of MDL 63,246, and examination of synthetic Microbiological importance. antimicrobial Chemothermy.39: 1580-1588.; Misiek, M.M., T.A.Pursiano, F.Leitner, and K.E.Prime 1973.Microbiological protocols of genetic engineering a, BL-S: 7- (phthalic acid-3-40) 3-4-carboxyethyl-3-4-3-4-carboxyethyl-3-4-carboxyethyl-3-4-3-5-carboxyethyl-4-carboxyethyl-4-3-4-carboxyethyl-3-4-3-4-3-4-3-4-one.

Preparation of Compounds

The compounds were dissolved in 2% tween 80 for oral administration or in 0.9% NaCl solution for intravenous administration. Compounds were administered 1 hour after bacterial inoculation. Vancomycin or ampicillin was used as a control.

Efficacy model

Evaluation was performed using male or female ICR mice weighing 22 + -2 g from MDS Pharma Services. Food and water were given freely. Experiments were performed using 6 mice per group weighing 22 ± 2 g. Intraperitoneal inoculation of mice with a solution containing 4X 10⁴0.5ml brain heart infusion (Difco) of CFU Staphylococcus aureus Smith containing 5% mucin (Sigma). Mortality was recorded once daily for 7 days after bacterial inoculation.

Although the invention has been described and illustrated herein with reference to a number of different specific materials, processes, and examples, it is to be understood that the invention is not limited to the specific materials, combinations of materials, and processes selected for this purpose. Those skilled in the art will appreciate numerous variations in such details.

Claims

1. A compound of formula (IA):

wherein:

represents a bond, is a double or single bond;

R¹selected from the group consisting of-S-alkyl, -S-substituted alkyl(heteroaryl) alkyl, hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cycloalkylalkyl, halo, and substituted alkylthio;

R⁶selected from hydrogen, alkyl, substituted alkyl, (amido) alkyl, (carbamoyl) alkyl, alkoxycarbonyl, (alkoxycarbonyl) alkyl, (alkoxycarbonylamino) alkyl, or-N (R)⁶) -the fragment is part of an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyamidine structure;

R⁷selected from the group consisting of hydrogen and alkyl;

R⁹may be mono-or multiply substituted on the same or different carbons in the ring, independently selected from hydrogen, substituted alkyl, halo, substituted alkenyl, (heteroaryl) alkenyl and-S (O)_qR¹³Group of (I), wherein q is an integer equal to 0, 1 or 2, R ¹³Selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, and substituted heterocyclyl;

wherein m is 0-2;

wherein t is 0-3;

or a pharmaceutically acceptable salt thereof;

with the proviso that the following compounds are excluded:

a compound of the formula (I), whereinIs a single bond, each R⁹Is hydrogen;

a compound of the formula (I), whereinIs a single bond, R⁹Is a substituted alkyl group having a single substituent other than halo, oxygen, hydroxyl, primary amine, amine (secondary alkyl substituted with the above alkyl), amine (tertiary alkyl substituted with the above alkyl), sulfur, -SH, phenyl or- (CH)₂)_nNR 'R ", where n is an integer from 1 to 8, R' and R" are hydrogen or alkyl; and

a compound of the formula (I), whereinIs a single bond, R⁹Is a halo group.

2. The compound of claim 1 wherein at R¹In the definition, -S-alkyl is SMe and-S-substituted alkyl is-S- (2-hydroxyethyl).

3. The compound of claim 1, wherein the nitrogen-containing ring in formula (IA) is selected from:

4. a compound selected from the group consisting of:

4-fluoro-4-propyl-piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide; and

4-fluoro-4-propyl-pyrrolidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

5. A compound of formula (IB):

wherein:

R⁶is H, alkyl or hydroxyalkyl;

R⁷is H or alkyl;

R⁹may be mono-or multiply-substituted on the same or different carbons of the ring, and is selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted oxygen, substituted nitrogen, halogen, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵And branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is H or alkyl;

m is 1 or 2;

or a pharmaceutically acceptable salt thereof.

6. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

7. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of any one of claims 1-6.

8. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 for the manufacture of a medicament for the treatment of a microbial infection in a mammal.

9. Use according to claim 8, wherein the medicament is in oral, parenteral, transdermal, topical, rectal or intranasal administration form.

10. Use according to claim 8, wherein the medicament is formulated such that the compound is administered in an amount of 0.1 to 100mg/kg body weight/day.

11. A compound of formula (I):

wherein:

w is a nitrogen-containing ring:

R⁶selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, substituted alkyl, aminomethyl, -C (O) O-alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, - (amido) alkyl, (carbamoyl) alkyl, 5-alkyl- [1, 3]Dioxolen-2-one-4-yl-methyl, 5-alkyl- [1, 3 [ ]]Dioxol-2-one-4-yl-methoxy-carbonyl group, or-N (R)⁶) The fragment is an amidine, N-cyanoamidine, N-hydroxyamidine or N-alkoxyA moiety of an amidine structure;

R⁷is H or alkyl;

R⁹may be mono-or multiply-substituted on the same or different carbons in the ring, independently selected from hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkylalkyl, substituted alkenyl, substituted oxygen, substituted nitrogen, halogen, phenyl, substituted phenyl, alkylthio, substituted arylthio, heteroarylthioalkyl, heterocyclylthioalkyl, heteroarylthio, and heterocyclylthio, ═ CHCH ₂CH₃Azido, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵And branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is H or alkyl; alkoxyalkoxy, aryl, substituted aryl, alkenyl, substituted alkenyl and-S (O)_qR¹³Group of (I), wherein q is an integer equal to 0, 1 or 2, R¹³Selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl and substituted heterocyclyl, wherein no more than one-S (O) is present on the nitrogen-containing ring_qR¹³A group;

wherein t is 0-3;

or a pharmaceutically acceptable salt thereof;

with the proviso that the following compounds are excluded:

a compound of the formula (I), whereinIs a single bond, each R⁹Is hydrogen;

a compound of the formula (I), whereinIs a single bond, R⁹Is a substituted alkyl group having a single substituent other than halogeno, oxygen, hydroxyl, primary amine, amine (secondary alkyl group substituted with the above alkyl group), amine (tertiary alkyl group substituted with the above alkyl group)Sulfur, -SH, phenyl or- (CH)₂)_nNR 'R ", where n is an integer from 1 to 8, R' and R" are hydrogen or alkyl; and

a compound of the formula (I), whereinIs a single bond, R⁹Is a halo group.

12. A compound of formula (I A):

wherein:

represents a bond, is a double or single bond;

R⁷selected from the group consisting of hydrogen and alkyl;

R⁹may be mono-or multiply substituted on the same or different carbons in the ring, and is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, aryl, substituted aryl, alkenyl, substituted alkenyl, and-S (O) _qR¹³Group of (I), wherein q is an integer equal to 0, 1 or 2, R¹³Selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, and substituted heterocyclyl;

wherein m is¹＝0-2；

Wherein t is 0-3;

or a pharmaceutically acceptable salt thereof;

with the following conditions:

A. in the compounds of the formula (I), ifIs a single bond, and is,

m¹is a number of 0 or 1, and,

R⁶is hydrogen, alkyl, hydroxyalkyl, -C (O) O-alkylene-cycloalkyl, -C (O) O-alkylene-substituted alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-substituted aryl, -C (O) O-heteroaryl, -C (O) O-substituted heteroaryl, -C (O) O-heterocyclyl, -C (O) O-substituted heterocyclyl, - [ C (O) O ] O]_p-alkylene-heterocycle, - [ C (O) O]_p-alkylene-substituted heterocycles wherein p is 0 or1，

R⁷Selected from the group consisting of hydrogen and alkyl,

then R is¹Is not-S-alkyl;

B. in the compounds of the formula (I), ifIs a single bond, and is,

R⁷selected from the group consisting of hydrogen and alkyl,

R¹selected from the group consisting of-S-alkyl, -S-substituted alkyl, hydrogen, alkyl, substituted alkyl A group consisting of a alkenyl group, a substituted alkenyl group, an alkoxy group, a substituted alkoxy group and a halogeno group,

then at least one R⁹Is not hydrogen, alkyl, substituted alkyl, alkoxyalkoxy, cycloalkyl, substituted oxygen, substituted nitrogen, halo, phenyl, substituted phenyl, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵-alkylene-R^aWherein R is^aSelected from the group consisting of a fluorophenyl group or a chlorophenyl group, and branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is hydrogen or alkyl;

C. in the compounds of the formula (I), ifIs a single bond, and is,

R⁷selected from the group consisting of hydrogen and alkyl,

wherein:

Substituted oxygen represents the group-OR^dWherein R is^dAre alkyl, haloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alkenyl, cycloalkyl and substituted cycloalkyl,

13. The compound of any one of claims 11 or 12, wherein the nitrogen-containing ring in formulae (I) and (IA) is selected from:

14. a compound of formula (IB):

wherein:

R⁶is H, alkyl or hydroxyalkyl;

R⁷is H or alkyl;

m is 1 or 2;

or a pharmaceutically acceptable salt thereof.

15. A compound of formula (II):

wherein:

w is a nitrogen-containing ring:

R²⁰and R²¹Independently is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, cyano, alkylthio, substituted alkylthio, hydroxy, halo, or R²⁰And R²¹One is NOR⁷The other is absent, or R²⁰And R²¹One is ═ CH₂And the other is absent; or R²⁰And R²¹Together is cycloalkyl, aryl or heterocyclyl;

R⁷is H or alkyl;

R⁹May be mono-or multiply-substituted on the same or different carbons in the ring, independently selected from hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkylalkyl, substituted alkenyl, substituted oxygen, substituted nitrogen, halogen, phenyl, substituted phenyl, alkylthio, substituted arylthio, heteroarylthioalkyl, heterocyclylthioalkyl, heteroarylthio, heterocyclylthio, ═ CHCH₂CH₃Azido, - (CH)₂)_n-OH、-(CH₂)_n-NR⁴R⁵And branched isomers thereof, wherein n is an integer from 1 to 8 inclusive, R⁴And R⁵Is H or alkyl; alkoxyalkoxy, aryl, substituted aryl, alkenyl, substituted alkenyl and-S (O)_qR¹³Group of (I), wherein q is an integer equal to 0, 1 or 2, R¹³Selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl and substituted heterocyclyl, wherein no more than one-S (O) is present on the nitrogen-containing ring_qR¹³A group;

or a pharmaceutically acceptable salt thereof.

16. A compound selected from the group consisting of:

3- (3-cyclobutyl-propyl) -azetidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide; and

4- (2-cyclopropyl-ethyl) -piperidine-2-carboxylic acid [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propyl ] -amide.

17. The following compounds:

2- [ 2-chloro-1- (3, 4, 5-trihydroxy-6-methylsulfanyl-tetrahydro-pyran-2-yl) -propylcarbamoyl ] -5-propyl-azepane-1-carboxylic acid 5-methyl-2-oxo- [1, 3] dioxol-4-ylmethyl ester.

18. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of any one of claims 11-17.

19. Use of a compound according to any one of claims 11 to 17 or a pharmaceutical composition according to claim 18 for the manufacture of a medicament for the treatment of a microbial infection in a mammal.

20. Use according to claim 19, wherein the medicament is for the treatment of a microbial infection caused by any of the following bacterial pathogens: haemophilus influenzae, enterococcus faecalis, and enterococcus faecium.

21. Use according to claim 19 or 20, wherein the medicament is in oral, parenteral, transdermal, topical, rectal or intranasal administration form.

22. Use according to claim 19 or 20, wherein the medicament is formulated such that the compound is administered in an amount of 0.1 to 100mg/kg body weight/day.