HK1175181B

HK1175181B - Sultam derivatives

Info

Publication number: HK1175181B
Application number: HK13102636.1A
Authority: HK
Inventors: Kevin William Anderson; Paul Gillespie; Nicholas John Silvester Huby; Norman Kong; John Lawson Roberts; Pamela Loreen Rossman; Sung-Sau So
Original assignee: 霍夫曼-拉罗奇有限公司
Priority date: 2009-11-24
Filing date: 2010-11-19
Publication date: 2016-02-26

Description

Sultam derivatives

Technical Field

The present invention relates to compounds of formula 1 and pharmaceutically acceptable salts thereof,

wherein R is₂、R₃、R₄、R₅And R₆As defined herein.

These compounds have cytotoxic activity. Thus, they are useful in the treatment or control of proliferative disorders such as cancer, particularly solid tumors.

The invention also relates to compositions and unit dose formulations comprising the compounds of the invention, methods of preparing such compounds, and methods of using such compounds in the treatment of proliferative disorders, such as cancer, particularly solid tumors, and most particularly breast, lung, colon and prostate tumors.

Background

Various disease states are characterized by uncontrolled cellular proliferation and differentiation. These disease states encompass a variety of cell types and diseases, such as cancer, atherosclerosis, and restenosis.

The term cancer is used to describe a class of diseases characterized primarily by uncontrolled cell growth. Cancer is one of the leading causes of death in the world today and is predicted to become the leading cause of death in the next few years. By the time of 2030, new cancer cases exceeding 2000 million and at least 1300 million deaths will be diagnosed each year.

There are many different forms of cancer, and many of these require different forms of treatment. The major forms of current cancer therapy include surgery, radiation therapy, bone marrow transplantation, immunotherapy, anti-angiogenesis therapy, and therapy with cytotoxic agents (commonly referred to as chemotherapy). A large number of cytotoxic agents have been used in the past 70 years to treat cancer, including nitrogen mustards (such as nitrogen mustards and estramustine); anthracycline antibiotics (such as doxorubicin, daunorubicin, and demethoxydaunorubicin); platinum-containing compounds (such as cisplatin, carboplatin, and oxaliplatin); antimetabolites (such as dacarbazine, capecitabine, fludarabine, 5-fluorouracil, gemcitabine, methotrexate and pemetrexed); topoisomerase inhibitors (such as topotecan and irinotecan); tubulin polymerization inhibitors (such as vinblastine and vincristine (vincristine), and tubulin depolymerization inhibitors (such as paclitaxel and docetaxel).

Although a variety of cytotoxic agents are known and have achieved great success as therapeutic agents for the treatment of a variety of cancers, there is still a substantial unmet need for new therapeutic approaches and a need for new cytotoxic agents.

Disclosure of Invention

in the formula 1, the compound is shown in the specification,

wherein R is₂、R₃、R₄、R₅And R₆As defined herein.

These compounds have cytotoxic activity. Thus, they are useful in the treatment or control of proliferative disorders, such as cancer, in particular solid tumors.

Detailed Description

The term "alkenyl" as used herein refers to a straight or branched chain hydrocarbon group having at least one double bond and 2 to 6, preferably 2 to 4, carbon atoms. Examples of "alkenyl groups" are vinyl (vinyl) (ethenyl)), allyl, isopropenyl, 1-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-ethyl-1-butenyl, 3-methyl-2-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-methyl-3-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl and 5-hexenyl.

The terms "alkoxy" and "alkoxy" as used herein each refer to a group in which an alkyl group (as defined below) is attached to an oxygen atom. The term "lower alkoxy" refers to a group wherein a lower alkyl (as defined below) is attached to an oxygen atom. Typical lower alkoxy groups include methoxy, ethoxy, isopropoxy or propoxy, butyloxy (butyloxy), and the like.

The term "alkyl" as used herein refers to a straight or branched chain saturated hydrocarbon group having from 1 to about 20 carbon atoms, and in certain embodiments from 1 to about 7 carbon atoms. The term "lower alkyl" refers to an alkyl group having from 1 to 6 carbon atoms, and in certain embodiments from 1 to 4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, 2-dimethylpentyl, and sec-pentyl.

The term "alkynyl" as used herein refers to a straight or branched chain hydrocarbon group having at least one triple bond and 2 to 6, preferably 2 to 4, carbon atoms. Examples of "alkynyl groups" are ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl and 5-hexynyl.

The term "aryl" as used herein refers to a monocyclic or bicyclic aromatic hydrocarbon group, preferably containing from 6 to 10 ring carbon atoms. Preferred aryl groups include, but are not limited to, phenyl, naphthyl, tolyl, and xylyl.

The term "azole" as used herein refers to a 5-membered heteroaryl group (defined below) wherein at least one heteroatom (defined below) is nitrogen. An "oxadiazole" is an azole having three heteroatoms, two being nitrogen and one being oxygen. "triazole" is an azole having three heteroatoms and three of each nitrogen. "tetrazole" is an azole having four heteroatoms and four of which are nitrogen.

The term "carrier" as used herein refers to a pharmaceutically inert vehicle (e.g., solvent, suspending agent) used to deliver an active compound, e.g., a compound of the invention, to a patient.

The term "cycloalkenyl" as used herein refers to a stable monocyclic or polycyclic non-aromatic hydrocarbon radical which is unsaturated and which contains from 5 to 10 ring atoms. Examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl or cyclohexenyl.

The term "cycloalkyl" as used herein refers to a stable monocyclic or polycyclic, non-aromatic, saturated hydrocarbon radical containing from 3 to 10 ring atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, bicycloalkyl (including bicycloheptyl groups such as bicyclo [3.1.1] heptyl and bicyclo [2.2.1] heptyl).

The term "excipient" as used herein refers to a pharmaceutically inert substance.

The term "halogen" as used herein means fluorine, chlorine, bromine or iodine, preferably fluorine and chlorine.

The term "heteroaryl" as used herein refers to an aromatic monocyclic or bicyclic group containing at least one heteroatom. The term "heteroatom" as used herein refers to a ring atom that is nitrogen, oxygen, or sulfur. Preferred heteroaryl groups include, but are not limited to, thienyl, furyl, indolyl, pyrrolyl, pyridyl, pyrazinyl, oxazolyl, oxadiazolyl, thiazolyl, quinolinyl, pyrimidinyl, imidazolyl, triazolyl, and tetrazolyl. In the case of bicyclic heteroaryl groups, it is to be understood that the ring atoms of one ring may all be carbon and the other ring may contain heteroatoms.

The term "heterocycle" as used herein refers to a 4-8 membered monocyclic or bicyclic saturated or partially unsaturated non-aromatic group containing 1-3 heteroatoms. Examples of heterocycles include pyrrolidin-2-yl; pyrrolidin-3-yl; a piperidinyl group; morpholin-4-yl and the like. In the case of bicyclic heterocycles, it is understood that the ring atoms of one ring may all be carbon and the other ring may contain heteroatoms.

As used herein, the term IC₅₀By "is meant the concentration of a particular compound required to inhibit 50% of a particular measured activity. IC (integrated circuit)₅₀In particular, the measurements can be performed as described below.

As used herein, the term IC₉₀By "is meant the concentration of a particular compound required to inhibit 90% of a particular measured activity. IC (integrated circuit)₉₀In particular, as described belowThe measurement is performed.

As used herein, a compound is said to exhibit "cytotoxic activity" if the percentage of inhibition measured using the assay of example 36 is at least about 50%. In preferred embodiments, the percent inhibition measured using such an assay is at least about 75% and even more preferably at least about 95%.

As used herein, the term "pharmaceutically acceptable" (e.g., carriers, salts, esters, and the like) in reference to a compound means that the compound is pharmacologically acceptable and substantially non-toxic to a subject to whom the particular compound is administered.

In the context of the foregoing definition of "pharmaceutically acceptable", a "pharmaceutically acceptable salt" of a compound is a conventional acid addition salt or base addition salt that retains the biological effectiveness and properties of the compound and is formed from a suitable non-toxic organic or inorganic acid or base. Examples of acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, trifluoroacetic acid, and the like. Examples of base addition salts include those derived from ammonium hydroxide, lithium hydroxide, potassium hydroxide, sodium hydroxide, and quaternary ammonium hydroxides such as, for example, tetramethylammonium hydroxide. Chemical modification of a pharmaceutical compound (i.e., drug) to convert it to a salt is a well-known technique employed by pharmaceutical chemists to obtain improved physical and chemical stability, hygroscopicity, flowability, and solubility of compounds. See, e.g., Ansel et al, Pharmaceutical Dosage Forms and drug Delivery Systems (1995) pgs.456-457.

In the context of the foregoing definition of "pharmaceutically acceptable", a "pharmaceutically acceptable ester" of a compound is a conventional ester of a compound, which contains a hydroxyl or carboxyl group; the esters retain the biological effectiveness and properties of the compounds and are capable of being cleaved in vivo (in the organism) to the corresponding active alcohol or carboxylic acid, respectively.

The term "substituted" as used herein to describe any of the above-described chemical groups (e.g., substituted alkyl, substituted aryl, substituted heteroaryl) refers to a chemical group wherein 1 to 5 (preferably 1 to 3) hydrogen atoms have been independently replaced by a substituent.

The term "unit dose formulation" as used herein refers to a pharmaceutical formulation (e.g., tablet, capsule) containing an active agent (e.g., a compound of the invention) in a stable form and capable of being administered to a patient in a single dose.

The present invention relates to compounds of formula 1 and pharmaceutically acceptable salts of such compounds,

in the formula 1, the compound is shown in the specification,

wherein the compound exhibits cytotoxic activity and:

R₂is hydrogen or dimethylamino;

R₃is hydrogen;

R₄is cycloalkyl optionally substituted with alkyl, or

Benzyl optionally substituted with halogen;

or R₃And R₄Together with the nitrogen atom to which they are attached form a piperidinyl group substituted with one or more substituents independently selected from the group consisting of: hydroxy and phenyl optionally substituted with halogen;

R₅is2, 2-dimethylpropyl or benzyl optionally substituted with halogen; and

R₆is phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, alkoxy optionally substituted by halogen, alkyl, optionally substituted by halogen, cyanoA phenoxy group of a group or an alkyl group and a phenyl group optionally substituted with an alkyl group or a halogen.

In one embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₂Is hydrogen.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₂Is dimethylamino.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₄Is cycloalkyl optionally substituted with alkyl.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₄Is benzyl optionally substituted with halogen.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₃And R₄Together with the nitrogen atom to which they are attached form a piperidinyl group substituted with one or more substituents independently selected from the group consisting of: hydroxyl and phenyl optionally substituted with halogen.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₅Is2, 2-dimethylpropyl.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₅Is benzyl optionally substituted with halogen.

In another embodiment of the invention, the compound is a compound that exhibits cytotoxic activityA compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R₆Is phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, alkoxy optionally substituted with halogen, alkyl, phenoxy optionally substituted with halogen, cyano or alkyl and phenyl optionally substituted with alkyl or halogen.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₄Is bicycloheptyl substituted three times with methyl.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₄Is adamantyl.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, wherein R is a compound showing cytotoxic activity₄Is bicyclo [3.1.1]Hept-3-yl.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, which exhibits cytotoxic activity, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

R₆is phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, alkoxy optionally substituted with halogen, alkyl, phenoxy optionally substituted with halogen, cyano or alkyl and phenyl optionally substituted with alkyl or halogen.

Examples of such compounds include:

2- (3- { [ (5-chloro-2-hydroxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-iodo-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-ethoxy-benzyl) -amino]Methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (5-isopropyl-2-methoxy-benzyl) -amino]-methyl } -benzyl) -1,1-1 lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (2-difluoromethoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- [3- ({ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - [2- (4-fluoro-phenoxy) -benzyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-p-tolyloxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-Dimethylaminones)Ylmethyl-3, 3-dimethyl-butyl) - (4' -fluoro-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-hydroxy-4-methylbenzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-hydroxy-5-methyl-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2' -methyl-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-fluoro-6-phenoxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2,3, 6-trifluoro-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (6-bromo-2-hydroxy-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (6-chloro)-2-fluoro-3-methyl-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethylbutyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ [2- (4-cyano-phenoxy) -benzyl]- ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (3-bromo-2, 6-difluoro-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,5S) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide; and

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide.

In another embodiment of the present invention, the compound is a compound of formula 1 or a pharmaceutically acceptable salt thereof, which exhibits cytotoxic activity, wherein

R₂Is hydrogen;

R₃is hydrogen;

R₄is adamantyl;

R₅is2, 2-dimethylpropyl; and

R₆is optionally substituted with 1-3 substituents independently selected fromPhenyl group of substituents of group (d): halogen, hydroxy, alkoxy optionally substituted with halogen, alkyl, phenoxy optionally substituted with halogen, cyano or alkyl and phenyl optionally substituted with alkyl or halogen.

Examples of such compounds include:

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-1-ylamide;

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-2-ylamide; and

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-1-ylamide.

R₂is hydrogen;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

R₆is benzodioxolyl.

An example of such a compound is 2- (3- { [ benzo [1,3 ]]Dioxolen-4-ylmethyl- ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt.

R₂is hydrogen;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is benzyl optionally substituted with halogen; and

Examples of such compounds include:

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide;

2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide;

2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide; and

2- (3- { [ (2, 3-difluoro-6-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acidAcid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide.

R₂Is hydrogen;

R₃and R₄Together with the nitrogen atom to which they are attached form a piperidinyl group substituted with one or more substituents independently selected from the group consisting of: hydroxy and phenyl optionally substituted with halogen;

R₅is2, 2-dimethylpropyl; and

Examples of such compounds include:

[2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- (4-phenyl-piperidin-1-yl) -methanone; and

[2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- [4- (4-chloro-phenyl) -4-hydroxy-piperidin-1-yl group]-a ketone.

R₂Is hydrogen;

R₃is hydrogen;

R₄is optionally selected fromBenzyl substituted with halogen;

R₅is2, 2-dimethylpropyl; and

An example of such a compound is 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid 4-chloro-benzylamide.

R₂Is hydrogen;

R₃is hydrogen;

R₄is bicyclo [2.2.1] optionally substituted with lower alkyl]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

R₆is phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, alkoxy optionally substituted with halogen, alkyl, phenoxy optionally substituted with halogen, cyano or alkyl and aryl optionally substituted with alkyl or halogen.

Examples of such compounds include:

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((S) -1,7, 7-trimethyl-bicyclo [ 2.2.1)]Hept-2-yl) -amide;

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl)(S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl-amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,4S) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amide; and

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,4R) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amide.

R₂is dimethylamino;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

An example of such a compound is 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide hydrochloride.

R₂is hydrogen or dimethylamino;

R₃is hydrogen;

R₄is a bicyclic or tricyclic cycloalkyl group having 7 to 10 ring atoms, optionally substituted with lower alkyl, or 4-chlorobenzyl;

or R₃And R₄Together with the nitrogen atom to which they are attached form 4-phenyl-piperidine or 4- (4-chloro-phenyl) -4-hydroxy-piperidine;

R₅selected from the group consisting of: 2, 2-dimethylpropyl; a benzyl group; 3-chlorobenzyl; and 4-chlorobenzyl;

R₆is composed of

R₈Selected from the group consisting of: bromine; a cyanophenoxy group; a difluoromethoxy group; an ethoxy group; fluorine; a hydroxyl group; iodine; a methoxy group; phenyl optionally substituted with one substituent which is halogen or lower alkyl; and a phenoxy group optionally substituted with one substituent selected from the group consisting of: halogen, cyano and lower alkyl;

R₉selected from the group consisting of: hydrogen; a methoxy group; a methyl group; fluorine; and bromine;

or R₈And R₉Together are-O-CH₂-O-；

R₁₀Is hydrogen or methyl;

R₁₁selected from the group consisting of: chlorine; fluorine; hydrogen; isopropyl group; and a methyl group; and

R₁₂selected from the group consisting of: chlorine; fluorine; hydrogen; a hydroxyl group; a methoxy group; and a phenoxy group.

R₂is hydrogen;

R₃is hydrogen;

R₆is composed of

R₉selected from the group consisting of: hydrogen; a methoxy group; and bromine;

or R₈And R₉Together are-O-CH₂-O-；

R₁₀Is hydrogen or methyl;

R₁₂selected from the group consisting of: chlorine; fluorine; hydrogen; and a hydroxyl group.

R₂is hydrogen or dimethylamino;

R₃is hydrogen;

R₆is composed of

R₈Selected from the group consisting of: bromine; a difluoromethoxy group; an ethoxy group; fluorine; iodine; phenyl substituted with one substituent which is halogen or lower alkyl; and a phenoxy group optionally substituted with one substituent selected from the group consisting of: halogen, cyano and lower alkyl;

R₉selected from the group consisting of: hydrogen; a methoxy group; a methyl group; (ii) a methylenedioxy group; fluorine; and bromine;

or R₈And R₉Together are-O-CH₂-O-；

R₁₀Is hydrogen;

R₁₁is hydrogen; and

R₂Is hydrogen or dimethylamino;

R₃is hydrogen;

R₅selected from the group consisting of: 2, 2-dimethylpropyl; a benzyl group; 3-chlorobenzyl; and 4-chlorobenzyl; and

R₆selected from the group consisting of:

2-bromo-6-hydroxy-3-methoxyphenyl;

6-chloro-2-fluoro-3-methoxyphenyl;

6-chloro-2-fluoro-3-methylphenyl;

5-chloro-2-hydroxyphenyl;

2- (4-cyanophenoxy) -phenyl;

2, 6-difluoro-3-bromophenyl;

2, 3-difluoro-6-methoxyphenyl;

2-difluoromethoxyphenyl;

2-ethoxyphenyl;

4' -fluoro-biphenyl-2-yl;

2-fluoro-6-phenoxyphenyl;

2- (4-fluorophenoxy) -phenyl;

2-hydroxy-4-methylphenyl;

2-hydroxy-5-methylphenyl;

2-iodophenyl;

5-isopropyl-2-methoxyphenyl;

2' -methyl-biphenyl-2-yl;

2, 3-methylenedioxyphenyl;

2- (4-methylphenoxy) -phenyl; and

2,3, 6-trifluorophenyl group.

The compounds of the invention have cytotoxic activity. Thus, they are useful in the treatment or control of cell proliferative disorders, such as cancer, particularly solid tumors, and most particularly breast, lung, colon and prostate tumors.

The compounds of the invention may exist in the form of racemic mixtures, diastereomeric mixtures, mixtures in proportions (scalemic mixtures) or isolated stereoisomers. Stereoisomers may be separated by known separation methods, for example by chromatography.

The compounds of the present invention may exhibit tautomerism or structural isomerism. The present invention is intended to encompass any tautomeric or structurally isomeric form of the compounds of the invention, or mixtures of such forms, and is not limited to any one tautomeric or structurally isomeric form described in the above formula.

The present invention also relates to a composition comprising a therapeutically effective amount of a compound of formula 1 exhibiting cytotoxic activity or a pharmaceutically acceptable salt thereof and a carrier.

The invention also relates to unit dose formulations comprising a therapeutically effective amount of a compound of formula 1, or a pharmaceutically acceptable salt thereof, which exhibits cytotoxic activity and a carrier.

A therapeutically effective amount of a compound of the invention is an amount of the compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. The therapeutically effective amount or dosage may vary within wide limits and may be determined in a manner known in the art. Such dosages will be adjusted to the individual requirements in each particular case (including the particular compound being administered, the route of administration, the disease being treated and the patient being treated). Generally, in the case of oral or parenteral administration of the compound of the present invention to an adult human weighing about 70kg, a daily dose of about 10mg to about 10,000mg, preferably about 200mg to about 1,000mg, should be appropriate, although the upper limit may be exceeded when specified. The daily dose may be administered in a single or divided dose, or for parenteral administration, in the form of a continuous infusion.

The invention also relates to processes for preparing the compounds of the invention. The compounds of the present invention may be prepared by any conventional method. Suitable methods for synthesizing these compounds are provided in the examples.

In one embodiment, the present invention therefore relates to a process for the preparation of a compound as described hereinbefore, comprising the steps of:

(i) reacting a compound of formula 2

Conversion to a compound of formula 3

(ii) Reacting a compound of formula 3 to form a compound of formula 4

(iii) Condensing a compound of formula 4 with a compound of formula 5

To give a compound of formula 6

(iv) Cleaving the ester group of the compound of formula 6 and reacting the resulting carboxylic acid with a compound of formula HNR₃R₄To give a compound of formula 7

(v) Converting the compound of formula 7 to the compound of formula 8

(vi) Condensing a compound of formula 8 with a compound of formula 9

To give a compound of formula 10

(vii) Converting the compound of formula 10 to the compound of formula 1

Wherein

R₁Is lower alkyl and

R₂、R₃、R₄、R₅and R₆As described hereinabove.

Another embodiment of the present invention relates to a compound as described hereinbefore, prepared according to said process.

In another embodiment, the invention also relates to a compound as described hereinbefore for use as a therapeutically active substance.

Another embodiment of the present invention relates to the use of a compound as described hereinbefore for the treatment or control of cell proliferative disorders, in particular breast, colon, lung and prostate tumors.

Another embodiment of the present invention also relates to the use of a compound as described hereinbefore for the preparation of a medicament for the treatment or control of cell proliferative disorders, in particular breast, colon, lung and prostate tumors.

A further embodiment of the present invention relates to compounds as described hereinbefore for the treatment or control of cell proliferative disorders, in particular breast, colon, lung and prostate tumors.

Another embodiment of the invention also relates to a method for the treatment or control of cell proliferative disorders, in particular breast, colon, lung and prostate tumors, which comprises administering an effective amount of a compound as described hereinbefore.

Preferably, the compound of formula 1 can be prepared according to scheme 1 from compounds of formula 2, formula 5 and formula 9: wherein R is₂、R₃、R₄And R₅、R₆As defined above and R₁Is an alkyl group.

Scheme 1

One general route for synthesizing the compounds of the present invention is shown in scheme 1. According to the method, the compound of formula 2 is converted to the compound of formula 3, and then the compound of formula 3 is reacted to form the compound of formula 4. The compound of formula 4 is then condensed with the compound of formula 5 to provide the desired compound of formula 6. Then breaking the ester group of the compound of formula 6 and reacting the resulting carboxylic acid with a compound of formula HNR₃R₄Coupling of the amine(s) to afford the desired compound of formula 7. The compound of formula 7 is then converted to a compound of formula 8, which is condensed with a compound of formula 9 to provide a compound of formula 10. Converting the compound of formula 10 to the desired compound of formula 1.

The compounds of formula 2 are commercially available. The racemic acid DL-homocystine can be obtained, for example, from Sigma-Aldrich, St.Louis, MO, USA. The homochiral substances L-homocystine and D-homocystine are also available from Sigma.

The reaction of the conversion of the compound of formula 2 to the compound of formula 3 can be carried out using a variety of conditions well known in the art, for example, by treating the compound of formula 2 with thionyl chloride in an alcohol solvent such as, for example, methanol or ethanol at a temperature of 0 ℃ to 25 ℃ to give the corresponding ester (wherein R is R₁The group depends on the alcohol used to carry out the reaction). For example, when the reaction is carried out in ethanol, the resulting compound is a compound of formula 3, wherein R is₁Represents an ethyl group. Examples of conditions for this reaction can be found in the literature, e.g., a. kelleman et al, Biopolymers 2003,71, 686-.

The reaction of converting the compound of formula 3 to the compound of formula 4 can be carried out using a variety of conditions well known in the art, for example, by treating the compound of formula 3 with an oxidizing agent (e.g., chlorine) in water or an alcohol solvent (e.g., methanol), followed by treatment with a tertiary alkylamine (e.g., triethylamine) at a temperature of-5 ℃ to 25 ℃. Examples of conditions for this reaction can be found in the literature, for example, G.Luisi et al. Archiv der Pharmazie 1993,326, 139-141; R.J.Cherney et al.J.Med.chem.2004,47, 2981-2983; and Z.Chen et al.Bioorg.Med.chem.Lett.2001,11, 2111-2115.

The reaction of a compound of formula 4 with a compound of formula 5 (the availability and preparation of which are discussed below), wherein X is a suitable leaving group such as a halide (e.g., bromo, chloro, iodo), an alkyl sulfonate, or an aryl sulfonate group (e.g., a mesylate or tosylate group), to form a compound of formula 6, can be carried out using a variety of conditions well known to those of ordinary skill in the art. For example, the compound of formula 4 may be reacted with the compound of formula 5 in the presence of a suitable base such as a metal carbonate (e.g., potassium carbonate, cesium carbonate, sodium carbonate or lithium carbonate, preferably potassium carbonate) or a metal hydride (e.g., sodium hydride or potassium hydride) in a suitable solvent such as N, N-dimethylformamide and/or tetrahydrofuran. The reaction may be carried out between about 0 ℃ and room temperature, preferably at room temperature. As another example, a compound of formula 4 may be reacted with a compound of formula 5 (where X is a hydroxyl group) in the presence of a trisubstituted phosphine such as triphenylphosphine or a triphenylphosphine polymer, a coupling agent such as 1, 2-diazenedicarboxylic acid 1, 2-diethyl ester, in an inert solvent such as tetrahydrofuran at a temperature between 0 ℃ and about room temperature, preferably at room temperature. Examples of conditions for this reaction can be found in the literature, for example, r.j.cherney et al.bioorg.med.chem.lett.2006,16, 1028-1031; and R.J.Cherneyeet al.J.Med.chem.2004,47, 2981-2983.

Hydrolysis of a compound of formula 6 (wherein R1 represents lower alkyl) to the corresponding carboxylic acid of formula 6 (wherein R1 represents hydrogen) can be carried out using conditions well known in the art of Organic Synthesis, wherein various conditions are summarized in "Protective Groups in Organic Synthesis" [ t.w.greene and p.g.m.wuts,2^nd Edition,John Wiley & Sons,N.Y.1991]In (1). For example, in which R₁In the case where methyl or ethyl is indicated, the reaction may conveniently be carried out by the addition of one equivalent of an alkali metal hydroxide such as potassium hydroxide, sodium hydroxide or lithium hydroxide, preferably lithium hydroxideBy treating the compound in a suitable solvent, such as a mixture of tetrahydrofuran, methanol and water. The reaction may be carried out at a temperature between about 0 ℃ and about room temperature (preferably at about room temperature). As another example, in which R₁In the case of a group which represents a group which can be cleaved under acidic conditions, such as a t-butyl group, the ester may be treated with a strong inorganic acid, for example, a hydrohalic acid such as hydrogen chloride or hydrogen bromide, or a strong organic acid, for example, an alkyl halide carboxylic acid such as trifluoroacetic acid, or the like. The reaction is conveniently carried out in the presence of an inert organic solvent such as dichloromethane and at a temperature between about 0 ℃ and about room temperature, preferably at room temperature. As a final, but not limiting example, in which R₁Representing the case where the group which can be cleaved by catalytic hydrogenation is present, and further conditions are satisfied, i.e. the rest of the molecule is stable to such conditions, the reaction can be carried out by hydrogenation in the presence of a noble metal catalyst such as Pd/C and an inert solvent, for example an alcohol such as ethanol, at about room temperature and at atmospheric pressure.

According to scheme 1, carboxylic acids derived from compounds of formula 6 (wherein R₁Representing hydrogen) and the structure HNR₃R₄The coupling of the amines of (the availability and preparation methods of) are discussed below can be accomplished using methods well known to those of ordinary skill in the art. For example, the conversion can be by a carboxylic acid of formula 6 (wherein R is₁Representing hydrogen) or a suitable derivative thereof (such as an activated ester) with a structural HNR₃R₄Is carried out in the presence of a coupling agent (various examples are well known in the field of peptide chemistry), if necessary. The reaction is conveniently carried out by using the structure HNR₃R₄In the presence of a suitable base (such as diisopropylethylamine), a coupling agent (such as O- (benzotriazol-1-yl) -1,1,3, 3-tetramethyluronium hexafluorophosphate or TSTU), and optionally an additional reaction rate increasing substance (such as 1-hydroxybenzotriazole or 1-hydroxy-7-azabenzotriazole) in an inert solvent (such as a chlorinated hydrocarbon (e.g., dichloromethane) or N, N-dimethylformamide or N-methylpyrrolidone) at between about 0 ℃ and about room temperatureTemperature (preferably at room temperature) treatment of the carboxylic acid of Structure 6 (wherein R is₁Represents hydrogen). Alternatively, the reaction may be carried out by reacting a carboxylic acid of formula 6 (wherein R is₁Representing hydrogen) into an activated ester derivative (such as an N-hydroxysuccinimide ester), which is subsequently reacted with a structural HNR₃R₄Or a corresponding acid addition salt. The reaction sequence may be carried out by reacting a carboxylic acid of formula 6 (wherein R is₁Representing hydrogen) with N-hydroxysuccinimide or 1-hydroxybenzotriazole in the presence of a coupling agent such as N, N '-dicyclohexylcarbodiimide or 1-ethyl-3- (3' -dimethylaminopropyl) carbodiimide in an inert solvent such as tetrahydrofuran or dichloromethane at a temperature between about 0 ℃ and room temperature. The resulting N-hydroxysuccinimide ester or 1-hydroxybenzotriazole ester is then treated with the structural HNR in the presence of a base such as an organic base (e.g., triethylamine or diisopropylethylamine, etc.) in a suitable solvent such as N, N-dimethylformamide at about room temperature₃R₄Or a corresponding acid addition salt.

The conversion of the nitrile of formula 7 to the aldehyde of formula 8 can be carried out using any conventional method. The nitrile may be hydrogenated, for example, in the presence of Raney nickel (Raney nickel) in an inert solvent such as aqueous formic acid at about 100 ℃. Examples of precise conditions suitable for carrying out such reactions can be found in the literature, for example, a. tanaka et al.j.med.chem.1998,41, 2390-; or R.J.Sundberg et al.J.Heterocyclic chem.1988,25, 129-137. Alternatively, the nitrile may be treated with diisobutylaluminum hydride in an inert solvent (such as toluene or a mixture of dichloromethane and hexane) at a temperature between about-5 ℃ and about 0 ℃. Examples of precise conditions suitable for carrying out such reactions can be found in the literature, for example, e.g. fischer WO 2007147547; T.Komatsu et al.Bioorg.Med.chem.2007,15, 3115-; R.E.Gawley et al.J.org.chem.2007,72, 2187-2191; or C.Hardouin et al.J.Med.chem.2007,50, 3359-3368.

The reaction of the aldehyde of formula 8 with the amine of formula 9 (the preparation method of which is described below) can be carried out using one of a variety of well-known reactions in a process known as reductive amination. For example, an aldehyde of formula 8 can be treated with an amine of formula 9 to provide an intermediate imine, which can be reduced to provide a compound of formula 10. The reduction may be carried out by hydrogenation under noble metal catalysed conditions or may be carried out by treating the imine with a reducing agent such as sodium borohydride or sodium cyanoborohydride or preferably sodium triacetoxyborohydride. The imine formation and reduction may be performed as two separate steps, or they may be combined into a single step. The one-step approach is convenient and well known to those of ordinary skill in the art of organic synthesis. An overview of this reaction, focusing in particular on the use of sodium triacetoxyborohydride as reducing agent, has recently been published (a.f. abdel-magic and s.j. mehrman org. process res.dev.2006,10,971-. The reaction is conveniently carried out by treating the aldehyde of formula 8 with an amine of formula 9 in an inert solvent such as a halogenated hydrocarbon (e.g. dichloromethane or 1, 2-dichloroethane) in the presence of an optional additional water absorbing agent such as molecular sieve at about room temperature. A reducing agent such as (sodium cyanoborohydride or preferably sodium triacetoxyborohydride) is added either simultaneously with the aldehyde of formula 8 being combined with the amine of formula 9, or after an interval, such as about 1 hour. Examples of conditions that can be used for this reaction can be found in the literature, e.g., w.sallem et al.bioorg.med.chem.2006,14,7999-one 8013; WO 2006014133; e.bogatcleva et al.j.med.chem.2006,49, 3045-chose 3048; and d.h.boschelli et al.j.med.chem.2004,47, 6666-.

The reaction of a secondary amine of formula 10 with an aldehyde of formula 13, the availability and preparation of which are discussed below, is another example of reductive amination (of an aldehyde of formula 13). The reaction can be carried out under reaction conditions similar to those described above for the conversion of the aldehyde of formula 8 to the secondary amine of formula 10.

In addition to the methods outlined in scheme 1, the compounds of formula 1 can be prepared by the methods shown in scheme 2.

Scheme 2

According to this method, the intermediate of formula 6 (as described in scheme 1) can be converted to the aldehyde of formula 11. Reductive amination of the aldehyde affords a secondary amine of formula 12. Reductive alkylation of the amine with an aldehyde of formula 13 affords a tertiary amine of formula 14. Hydrolyzing the ester, followed by reaction with a compound of formula HNR₃R₄To give the compound of formula 1.

The conversion of the nitrile of formula 6 to the aldehyde of formula 11 can be carried out using any conventional method. The nitrile may be hydrogenated, for example, in the presence of raney nickel in an inert solvent such as aqueous formic acid at about 100 ℃. Examples of precise conditions suitable for carrying out such reactions can be found in the literature, for example, a. tanaka et al.j.med.chem.1998,41, 2390-; or R.J.Sundberg et al.J.Heterocyclic chem.1988,25, 129-137. Alternatively, the nitrile may be treated with diisobutylaluminum hydride in an inert solvent (such as toluene or a mixture of dichloromethane and hexane) at a temperature between about-5 ℃ and about 0 ℃. Examples of precise conditions suitable for carrying out such reactions can be found in the literature, for example, e.g. fischer WO 2007147547; T.Komatsu et al.Bioorg.Med.chem.2007,15, 3115-; R.E.Gawley et al.J.org.chem.2007,72, 2187-2191; or C.Hardouin et al.J.Med.chem.2007,50, 3359-3368.

The reaction of the aldehyde of formula 11 with the amine of formula 9, the preparation method of which is described below, can be carried out using one of a variety of well-known reactions in a process known as reductive amination. For example, an aldehyde of formula 11 can be treated with an amine of formula 9 to provide an intermediate imine, which can be reduced to provide a compound of formula 12. The reduction may be carried out by hydrogenation under noble metal catalysed conditions or may be carried out by treating the imine with a reducing agent such as sodium borohydride or sodium cyanoborohydride or preferably sodium triacetoxyborohydride. The imine formation and reduction may be performed as two separate steps, or they may be combined into a single step. The one-step approach is convenient and well known to those of ordinary skill in the art of organic synthesis. An overview of this reaction, focusing in particular on the use of sodium triacetoxyborohydride as reducing agent, has recently been published (a.f. abdel-magic and s.j. mehrman org. process res.dev.2006,10,971-. The reaction is conveniently carried out by treating the aldehyde of formula 11 with an amine of formula 9 in an inert solvent such as a halogenated hydrocarbon (e.g., dichloromethane or 1, 2-dichloroethane) in the presence of an optional additional water absorbing agent such as a molecular sieve at about room temperature. A reducing agent, such as sodium cyanoborohydride or preferably sodium triacetoxyborohydride, is added either simultaneously with the combination of the aldehyde of formula 11 and the amine of formula 9, or after an interval, such as about 1 hour. Examples of conditions that can be used for this reaction can be found in the literature, e.g., w.sallem et al.bioorg.med.chem.2006,14,7999-one 8013; WO 2006014133; e.bogatcleva et al.j.med.chem.2006,49, 3045-chose 3048; and d.h.boschelli et al.j.med.chem.2004,47, 6666-.

The reaction of a secondary amine of formula 12 with an aldehyde of formula 13, the availability and preparation of which are described below, is another example of reductive amination (of an aldehyde of formula 13). The reaction may be carried out under reaction conditions similar to those described above for the conversion of the aldehyde of formula 11 to the secondary amine of formula 12.

Hydrolysis of the ester functional group in the tertiary amine of formula 14 can be carried out using conditions well known in the art of Organic Synthesis, a variety of which are summarized in "Protective Groups in Organic Synthesis" [ T.W.Greene and P.G.M.Wuts,2^nd Edition,John Wiley & Sons,N.Y.1991]In (1). For example, in which R₁Where methyl or ethyl is indicated, the reaction may conveniently be effected by treating the compound with one equivalent of an alkali metal hydroxide such as potassium hydroxide, sodium hydroxide or lithium hydroxide, preferably lithium hydroxide, in a suitable solvent such as a mixture of tetrahydrofuran, methanol and water. The reaction may be carried out at a temperature between about 0 ℃ and about room temperature, preferably at about room temperature.

According to scheme 2, carboxylic acids of formula 15 with structural HNR₃R₄The coupling of the amines of (a) whose availability and preparation are described below can be achieved using methods well known to those of ordinary skill in the art.For example, the transformation may be by reaction of a carboxylic acid of formula 15 or a suitable derivative thereof (such as an activated ester) with the structural HNR₃R₄Is carried out in the presence of a coupling agent (various examples are well known in the field of peptide chemistry), if necessary. The reaction is conveniently carried out by using the structure HNR₃R₄Is carried out by treating the carboxylic acid of formula 15 in the presence of a suitable base such as diisopropylethylamine, a coupling agent such as (O- (benzotriazol-1-yl) -1,1,3, 3-tetramethyluronium hexafluorophosphate or TSTU), and optionally an additional reaction rate increasing substance such as 1-hydroxybenzotriazole or 1-hydroxy-7-azabenzotriazole, in an inert solvent such as a chlorinated hydrocarbon (e.g., dichloromethane) or N, N-dimethylformamide or N-methylpyrrolidinone at a temperature between about 0 ℃ and about room temperature, preferably at room temperature. Alternatively, the reaction may be carried out by converting the carboxylic acid of formula 15 to an activated ester derivative (such as an N-hydroxysuccinimide ester), followed by reaction with the structural HNR₃R₄Or a corresponding addition salt. This reaction sequence may be carried out by reacting the carboxylic acid of formula 15 with N-hydroxysuccinimide or 1-hydroxybenzotriazole in the presence of a coupling agent such as N, N '-dicyclohexylcarbodiimide or 1-ethyl-3- (3' -dimethylaminopropyl) carbodiimide in an inert solvent such as tetrahydrofuran or dichloromethane at a temperature between about 0 ℃ and room temperature. The resulting N-hydroxysuccinimide ester or 1-hydroxybenzotriazole ester is then treated with the structural HNR in the presence of a base such as an organic base (e.g., triethylamine or diisopropylethylamine, etc.) in a suitable solvent such as N, N-dimethylformamide at about room temperature₃R₄Or a corresponding acid addition salt.

Scheme 3 outlines an alternative method that can be used to convert intermediates of formula 12 to compounds of the invention of formula 1.

Scheme 3

According to the method shown in scheme 3, the ester group in the compound of formula 12 is cleaved to give the acid of formula 16. Then reacted with a compound of formula HNR₃R₄To give an amide of formula 17, followed by reductive alkylation of the secondary amine to give a compound of formula 1.

Hydrolysis of the ester functional group in the secondary amine of formula 12 can be carried out using conditions well known in the art of Organic Synthesis, a variety of which are summarized in "Protective Groups in Organic Synthesis" [ T.W.Greene and P.G.M.Wuts,2^nd Edition,John Wiley & Sons,N.Y.1991]In (1). For example, in which R₁Where methyl or ethyl is indicated, the reaction may conveniently be effected by treating the compound with one equivalent of an alkali metal hydroxide such as potassium hydroxide, sodium hydroxide or lithium hydroxide, preferably lithium hydroxide, in a suitable solvent such as a mixture of tetrahydrofuran, methanol and water. The reaction may be carried out at a temperature between about 0 ℃ and about room temperature (preferably at about room temperature).

According to scheme 3, carboxylic acids of formula 16 with the structure HNR₃R₄The coupling of the amines of (a) whose availability and preparation are described below can be achieved using methods well known to those of ordinary skill in the art. For example, the transformation can be by a carboxylic acid of formula 16 with the structural HNR₃R₄Is carried out in the presence of a coupling agent (various examples are well known in the field of peptide chemistry). The reaction is conveniently carried out by treating the carboxylic acid of formula 16 with the free base or hydrochloride salt of an amine of structure HNR3R4 in the presence of a suitable base such as diisopropylethylamine, a coupling agent such as O- (benzotriazol-1-yl) -1,1,3, 3-tetramethyluronium hexafluorophosphate or TSTU, and optionally an additional catalyst such as 1-hydroxybenzotriazole or 1-hydroxy-7-azabenzotriazole in an inert solvent such as a chlorinated hydrocarbon (e.g. dichloromethane) or N, N-dimethylformamide or N-methylpyrrolidone at a temperature between about 0 ℃ and about room temperature, preferably at room temperature. A variety of alternative coupling methods are known in the field of synthetic chemistry andand some of which have been outlined above (e.g. in the description of the conditions for the conversion of a compound of formula 15 to a compound of formula 1).

The reaction of the aldehyde of formula 13 (the availability and preparation of which are described below) with the amine of formula 17 can be carried out using one of the many well-known reactions in a process known as reductive amination. For example, an aldehyde of formula 13 can be treated with an amine of formula 17 to provide an intermediate imine, which can be reduced to provide a compound of formula 1. The imine formation and reduction may be performed as two separate steps, or they may be combined into a single step. The one-step approach is convenient and well known to those of ordinary skill in the art of organic synthesis. An overview of this reaction, focusing in particular on the use of sodium triacetoxyborohydride as reducing agent, has recently been published (a.f. abdel-magic and s.j. mehrman org. process res.dev.2006,10,971-. The reaction is conveniently carried out by treating the aldehyde of formula 13 with an amine of formula 17 in an inert solvent such as dichloromethane, optionally in the presence of an additional water absorbing agent such as molecular sieve, at about room temperature. A reducing agent, such as sodium cyanoborohydride or sodium triacetoxyborohydride, is added either simultaneously with the aldehyde of formula 13 being combined with the amine of formula 17, or after an interval, such as about 1 hour. Examples of conditions that can be used for this reaction can be found in the literature, e.g., w.sallem et al.bioorg.med.chem.2006,14,7999-one 8013; WO 2006014133; e.bogatcleva et al.j.med.chem.2006,49, 3045-chose 3048; and d.h.boschelli et al.j.med.chem.2004,47, 6666-.

Compounds of the invention (wherein R₂Representing dimethylamino) can also be prepared using the route outlined in scheme 4.

Scheme 4

According to this method, the compound of formula 18 ((3-hydroxymethyl-5-nitro-phenyl) -methanol) is mono-protected to give the compound of formula 19Wherein a represents a protecting group. The nitro group is then reduced to give aniline 20, and the nitrogen of the aniline is dimethylated to give the dimethylamino compound 21. Oxidation of the benzyl alcohol in 21 affords aldehyde 22. Removal of the protecting group affords alcohol 23. The alcohol is converted to benzyl bromide to afford intermediate 24. Intermediate 24 is then reacted with sultam 4 to provide compound 25, which is also a compound of formula 11, wherein R is₂Represents a dimethylamino group. Compound 25 can then be converted to the compound of formula 1 using the sequence of steps outlined in scheme 2.

Compounds of formula 18 are commercially available. For example, it may be purchased from Aldrich Chemical co, Milwaukee, WI, USA. The conversion of the diol of formula 18 to the mono-protected alcohol of formula 19 can be accomplished by any conventional method using one of a variety of protecting groups for alcohols well known in the art of organic synthesis. For example, various suitable Groups are outlined in "Protective Groups in Organic Synthesis" [ T.W.Greene and P.G.M.Wuts,2^nd Edition,John Wiley & Sons,N.Y.1991]In (1). One such suitable group is a t-butyldimethylsilyl (TBDMS) group. This can be easily introduced by treating the diol with tert-butyldimethylsilyl chloride in an inert solvent such as dichloromethane in the presence of imidazole at a temperature between about 0 ℃ and about room temperature. In this case, if one equivalent of tert-butyldimethylsilyl chloride is used, the reaction mixture will be converted into a statistical mixture of starting material, mono-protected diol and di-protected diol in a ratio of about 1:2: 1. The desired mono-protected compound of formula 19 is then isolated from the mixture using techniques that will be apparent to those of ordinary skill in the art of organic chemistry, such as by silica gel chromatography.

Reduction of the nitro Group in the compound of formula 19 to give the aniline derivative of formula 20 can be achieved using one of a number of methods well known to organic chemists, such as those outlined in "Comprehensive organic transformations: A Guide to Functional Group Preparations" [ R.C. Larock, VCHPublishes, Inc. New York,1989], for example page 412-415. A convenient method is to treat a solution of the compound of formula 19 with hydrogen in an inert solvent such as ethanol in the presence of a noble metal catalyst such as Pd/C at about room temperature.

The aniline of formula 20 can be converted to the dimethylamino derivative of formula 21 by treatment with formaldehyde in the presence of a reducing agent such as sodium cyanoborohydride and a lewis acid catalyst such as zinc chloride in an inert solvent such as methanol at a temperature of about room temperature. Formaldehyde can be produced by heating commercially available paraformaldehyde (so that it forms a gas) which can then be introduced into the reaction mixture. Examples of specific reaction conditions that may be used for such reactions may be found in the literature such as, for example, r.g. carter et al.wo 2008156656; m.ono et al.bioorg.med.chem.2008,16, 6867-; sueokaet al.us 6,288,061; and b.d. allison et al.j.med.chem.2006,49, 6371-6390.

The alcohol of formula 21 can then be converted to the aldehyde of formula 22 using one of a variety of well known reactions. Examples of precise conditions suitable for carrying out the oxidation of benzyl alcohol to benzaldehyde can be found in the literature, for example, j.s.yadav et al.tetrahedron 2004,60, 2131-; c.kuhakam et al. Commun.2006,36, 2887-2892; theeraldanon et al tetrahedron 2004,60, 3017-3035; zhao and a. thurkauf synth. commu.2001, 31, 1921-; A.W.white et al.J.Med.chem.2000,43, 4084-; J.Clayden et al tetrahedron 2004,60, 4399-4412; N.Maezaki et al.tetrahedron 2000,56, 7927-; A.P Combs et al.J.Med.chem.2006,49, 3774-3789; and R.M.Moriarty et al.J.org.chem.2004,68, 1890-.

The protected derivative of formula 22 can then be converted to the alcohol of formula 23 using one of a variety of conditions known to be useful for deprotecting the protected alcohol. These conditions depend on the nature of the protecting group, and an example of suitable conditions may be found in "Protective Groups in Organic Synthesis" [ T.W.Greene and P.G.M.Wuts,2^nd Edition,John Wiley & Sons,N.Y.1991]Is found in (1). For example, in the case where the protecting group is t-butyldimethylsilyl, the protecting group can be prepared by usingTreating the compound of formula 21 with aqueous acetic acid at about room temperature to remove the protecting group.

The conversion of the alcohol of formula 23 to benzyl bromide of formula 24 can be achieved by treating the compound of formula 23 with phosphorus tribromide or a mixture of N-bromosuccinimide and triphenylphosphine, or a mixture of carbon tetrabromide and triphenylphosphine, in an inert solvent such as an alkyl halide (e.g. dichloromethane or carbon tetrachloride or 1, 2-dichloroethane) or tetrahydrofuran or N, N-dimethylformamide, at a temperature between about 0 ℃ and the boiling point of the solvent, conveniently at about 0 ℃. Examples of precise conditions suitable for carrying out such substitution reactions can be found in the literature, for example, in s. rapperoseli et al. heterocycles 2008,75,1467-; W.Zeng and S.R.Chemler J.org.chem.2008,73, 6045-ion 6047; nicolaou, K.C.et al.tetrahedron2008,64, 4736-; and P.Sehnel et al.J.org.chem.2008,73, 2074-.

The reaction of the compound of formula 4 with the compound of formula 24 to produce the compound of formula 25 can be carried out using a variety of conditions well known to those of ordinary skill in the art. For example, a compound of formula 4 may be reacted with a compound of formula 24 in the presence of a suitable base, such as a metal carbonate (such as potassium carbonate, cesium carbonate, sodium carbonate or lithium carbonate, preferably potassium carbonate) or a metal hydride (such as sodium hydride or potassium hydride), in a suitable solvent, such as N, N-dimethylformamide and/or tetrahydrofuran. The reaction may be carried out at between about 0 ℃ and room temperature (preferably at room temperature). Examples of conditions for this reaction can be found in the literature, for example, r.j.cherney et al.bioorg.med.chem.lett.2006,16, 1028-1031; and R.J.Cherneyeet al.J.Med.chem.2004,47, 2981-2983.

As mentioned above, compounds of formula 25 are examples of compounds of formula 11, wherein R is₂Represents a dimethylamino group. The compound of formula 25 can be converted to the compound of the invention of formula 1 by following the sequence of steps depicted in scheme 2 for the conversion of the compound of formula 11 to the compound of formula 1, wherein R is₂Represents a dimethylamino group.

Availability of nitrile reagents of formula 5

Two nitrile reagents of formula 5 are commercially available. 3-cyano-benzyl bromide is available from Aldrich chemical company, Inc., Milwaukee, Wis., USA. 3-cyano-benzyl alcohol is available from TCI America, Portland, OR, USA.

Nitrile of formula 5 (wherein R₂Representing dimethylamino) can be conveniently prepared from 3-cyano-5-dimethylaminobenzoic acid (the synthesis of which is described in a. slasii et al. wo 2002068417). According to this method, the acid is treated with a chloroformate reagent (such as methyl chloroformate or ethyl chloroformate) in the presence of a base (such as triethylamine) in an inert solvent (such as toluene) at a temperature below about 0 ℃. The conditions described in j.a.price and d.s.tarbell org.syntheses 1957,37,20-23 may conveniently be used to effect this transformation. The resulting solution of mixed anhydride in tetrahydrofuran may then be added to a solution of sodium borohydride in a mixture of tetrahydrofuran and water at a temperature between about 0 ℃ and about room temperature to effect reduction to the alcohol of formula 5 (where R is₂Represents dimethylamino and X represents hydroxyl). This compound can be conveniently converted to a compound of formula 5 (wherein R is phosphorus tribromide or a mixture of N-bromosuccinimide and triphenylphosphine, as described above₂Represents dimethylamino and X represents bromine).

Preparation of an Alkaldehyde of formula 8

Scheme 5

The conversion of the reagent of formula 4 to the reagent of formula 8 can be performed using methods other than those outlined in scheme 1, as will be readily apparent to those of ordinary skill in the art of synthetic organic chemistry. For example, the conversion can be carried out by using a vinyl reagent of formula 26 (the availability and preparation of which are discussed below) shown in scheme 5. According to this method, alkylation of the compound of formula 4 with a reagent of formula 26 affords an intermediate of formula 27. Hydrolysis of the ester followed by amide coupling results in the formation of an intermediate of formula 28, which is then ozonolysis or oxidative cleavage to give the aldehyde of formula 8.

A compound of formula 4 with a compound of formula 26 (wherein R₂The reaction of a compound of formula 27 with a hydrogen or dimethylamino group and X is a suitable leaving group such as a halide (e.g., bromo, chloro, iodo), alkyl or aryl sulfonate (e.g., mesylate or tosylate)) may be carried out using a variety of conditions well known to those of ordinary skill in the art, and which are entirely similar to those described above in connection with the preparation of the compound of formula 6.

Hydrolysis of a compound of formula 27 to the corresponding carboxylic acid is carried out using conditions well known in the art of Organic Synthesis, a variety of which are summarized in "Protective Groups in Organic Synthesis" [ T.W.Greene and P.G.M.Wuts,2^nd Edition,John Wiley & Sons,N.Y.1991]And the conditions are also completely similar to those described above in connection with the hydrolysis of the compound of formula 6.

According to scheme 5, carboxylic acids of formula 27 (wherein R₁Representing hydrogen) and the structure HNR₃R₄The coupling of amines (the availability and preparation of which are discussed below) can be accomplished using techniques known to those of ordinary skill in the art and analogous to those described above for compounds derived from formula 6 (wherein R is₁Representing hydrogen) to those described for the preparation of the compound of formula 7.

A compound of formula 28 (wherein R₂Hydrogen or dimethylamino) to form the compound of formula 8 can be performed using a variety of conditions well known to those of ordinary skill in the art. For example, a compound of formula 28 can be reacted with a mixture of osmium tetroxide and sodium periodate in an inert solvent (such as a mixture of tetrahydrofuran and water or a mixture of acetone and water) at about room temperature. Examples of specific conditions for such reactions can be found in literature such as c.plisson et al.j.med.chem.2007,50, 4553-4560; J. U.S. Chung et al.Bioorg.Med.chem.2007,15, 6043-6053; or c.fluent et al.us20050131042. Alternatively, the compound of formula 28 can be treated with ozone gas in an inert solvent (such as a mixture of acetone and ethanol) or in dichloromethane at low temperatures (such as at about-10 ℃ or about-78 ℃), and the resulting ozonide can be treated with a reducing agent (such as zinc powder) in the presence of acetic acid or dimethyl sulfide without separation to give the aldehyde of formula 8. Examples of specific conditions for such reactions can be found in the literature, e.g., h.maeda et al.j.org.chem.2005,70,9693-; hikari et al J.Med. chem.2003,46, 3152-3161; or S.L.Swann et al.J.am.chem.Soc.2002,124, 13795-13805.

Bromomethyl-3-vinyl-benzene (compound of formula 26, wherein R₂Representing hydrogen and X representing bromine) can be conveniently prepared using the methods disclosed in a. naghipour et al polyhedron 2008,27, 1947-.

Bromomethyl-3-dimethylamino-5-vinyl-benzene (compound of formula 26, wherein R is₂Representing dimethylamino and X representing bromine) can be prepared in four steps from methyl 3-dimethylamino-5-hydroxybenzoate (the synthesis of which is described in r.w. rickards et al.wo 1984000750). According to this method, methyl 3-dimethylamino-5-hydroxybenzoate is converted to a triflate ester by reaction with trifluoromethanesulfonic anhydride in the presence of a base such as triethylamine, diisopropylethylamine, or pyridine in an inert solvent such as dichloromethane at between about-10 ℃ and about room temperature. The triflate is then subjected to an ethenylation reaction using vinyltributyltin in the presence of a palladium catalyst such as bis (triphenylphosphine) palladium (II) dichloride in an inert solvent such as dioxane at about 100 ℃ to give 3-dimethylamino-5-vinyl-benzoic acid methyl ester. Reduction of the ester by treatment with lithium aluminium hydride in an inert solvent such as diethyl ether or tetrahydrofuran at a temperature between about 0 ℃ and about 35 ℃ gives (3-dimethylamino-5-vinyl-phenyl) -methanol. As described above for the preparation of compounds of formula 5 (wherein X represents bromine), the compounds may be conveniently converted to compounds of formula 26 (wherein R is R) by treating them with phosphorus tribromide or a mixture of N-bromosuccinimide and triphenylphosphine₂Represents twoAmino and X represents bromo).

Preparation of amines of formula 9

Scheme 6

One general procedure for converting a compound of formula 29 (the availability and preparation of which are discussed below) to a compound of formula 9 is outlined in scheme 6. The reaction of the carboxylic acid of formula 29 to the alcohol of formula 30 can be accomplished using methods well known to those of ordinary skill in the art. For example, the conversion can be carried out by reaction of a carboxylic acid of formula 29 or a suitable derivative thereof (such as an ester group) with a suitable reducing agent (such as lithium aluminum hydride or lithium borohydride, preferably lithium borohydride and the like) in a suitable ether solvent (such as tetrahydrofuran and the like) at a temperature between about 0 ℃ and about room temperature (preferably at room temperature) to provide a compound of formula 30. The reaction may be carried out in the presence of trimethylchlorosilane. The amine of formula 30 is then converted to the protected derivative 31. Suitable protecting Groups for nitrogen include t-butyloxycarbonyl, benzyl, and the like, and a variety of suitable Groups can be found in t.greene and p.wuts, Protective Groups in Organic Synthesis,3rd.ed., John Wiley and Sons (1999). The amino group of the compound of formula 30 can be protected as a tert-butyl carbamate group using one of a variety of well-known reactions to provide a compound of formula 31 (wherein X represents a tert-butoxycarbonyl group). For example, the amine of formula 30 can be treated with di-tert-butyl dicarbonate or with 2- (Boc-oxyimino) -2-phenylacetonitrile (Boc-ON) in the presence of a base such as triethylamine or diisopropylethylamine in a solvent such as tetrahydrofuran or acetonitrile or a mixture of dioxane and water at about room temperature. References to methods useful for protecting amines (employing various other protecting groups) can be found in Greene and Wuts (cited above).

Activation of the alcohol of formula 31 with tosyl chloride followed by substitution with a dialkylamine, preferably dimethylamine, affords a compound of formula 33. The preparation of tosylate of formula 32 (where Q is p-tolyl) can be achieved by treating the alcohol of formula 31 with p-toluenesulfonyl chloride in the presence of a base such as triethylamine or diisopropylethylamine or pyridine in an inert solvent such as dichloromethane at a temperature between about 0 ℃ and about room temperature. The conversion of the tosylate of formula 32 to the amine of formula 33 is conveniently carried out by treating the compound of formula 32 with dimethylamine or dimethylamine hydrochloride in the presence of a base such as triethylamine or diisopropylethylamine or pyridine in an inert solvent such as dichloromethane at a temperature between about 0 ℃ and about room temperature. Alternatively, the reaction may be carried out using pyridine as a solvent in the absence of an additional solvent. Examples of specific conditions that may be used in this reaction can be found in J.Christoffs and A.Mann chem.Eur.J.2001,7, 1014-.

Deprotection of a compound of formula 33 to produce a compound of formula 9 can be accomplished using reactions well known to those of ordinary skill in the art. For example, the reaction of a compound of formula 33 with a protic acid, such as trifluoroacetic acid or hydrochloric acid, in a suitable inert solvent, such as dichloromethane or an ether solvent, such as diethyl ether and the like, at a temperature of from 0 ℃ to room temperature, preferably at room temperature. Examples of specific conditions for such reactions can be found in t.greene and p.wuts, Protective Groups in Organic Synthesis,3rd.ed., john wiley and Sons (1999).

The conversion of the reagent of formula 29 to the reagent of formula 9 can be performed using methods other than those outlined in scheme 6, as will be readily apparent to those of ordinary skill in the art of synthetic organic chemistry. For example, the conversion can be carried out by using the reaction sequence outlined in scheme 7.

Scheme 7

The compound of formula 29 may be conveniently reacted with a suitable protecting group for nitrogen. Suitable protecting groups for nitrogen include t-butyloxycarbonyl, benzyl, and the like. Other amino protecting Groups for compounds of formula 29 can be found in T.Greene and P.Wuts, Protective Groups in Organic Synthesis,3rd.Ed., John Wiley and Sons (1999). The amino group of the compound of formula 29 can be protected as tert-butyl carbamate using one of a variety of well-known reactions to give a compound of formula 34 (wherein X represents tert-butoxycarbonyl). For example, an amine of formula 29 can be treated with di-tert-butyl dicarbonate or with 2- (Boc-oxyimino) -2-phenylacetonitrile (Boc-ON) in the presence of a base such as triethylamine or diisopropylethylamine in a solvent such as tetrahydrofuran or acetonitrile or a mixture of dioxane and water at about room temperature. References to methods useful for protecting amines (employing various other protecting groups) can be found in Greene and Wuts (cited above).

The coupling of the carboxylic acid of formula 34 with dimethylamine to produce the compound of formula 35 according to scheme 7 can be accomplished using methods well known to those of ordinary skill in the art. For example, the conversion may be by reaction of a carboxylic acid of formula 34 or a suitable derivative thereof (such as an activated ester) with the structure HNMe₂Is carried out in the presence of a coupling agent (various examples are well known in the field of peptide chemistry), if necessary. The reaction is conveniently carried out by treating the carboxylic acid of formula 34 with dimethylamine (either as the free base or as the hydrochloride salt), in the presence of a suitable base such as diisopropylethylamine, a coupling agent such as O- (benzotriazol-1-yl) -1,1,3, 3-tetramethyluronium hexafluorophosphate or TSTU, and optionally in the presence of an additional reaction rate increasing substance such as 1-hydroxybenzotriazole or 1-hydroxy-7-azabenzotriazole. The reaction is carried out in an inert solvent such as a chlorinated hydrocarbon (e.g., dichloromethane) or N, N-dimethylformamide or N-methylpyrrolidone at a temperature between about 0 ℃ and about room temperature, preferably at room temperature. Alternatively, the reaction may be carried out by converting the carboxylic acid of formula 34 to an activated ester derivative (such as an N-hydroxysuccinimide ester) which is subsequently reacted with dimethylamine or a corresponding addition salt. This reaction sequence can be carried out by reacting a carboxylic acid of formula 34 with N-hydroxysuccinimide or 1-hydroxybenzotriazole in a coupling agent(s)Such as N, N '-dicyclohexylcarbodiimide or 1-ethyl-3- (3' -dimethylaminopropyl) carbodiimide) in an inert solvent such as tetrahydrofuran or dichloromethane at a temperature between about 0 ℃ and room temperature. The resulting N-hydroxysuccinimide ester or 1-hydroxybenzotriazole ester is then treated with dimethylamine or the corresponding acid addition salt in the presence of a base such as an organic base (e.g., triethylamine or diisopropylethylamine, etc.) in a suitable solvent such as N, N-dimethylformamide at about room temperature. Examples of specific reaction conditions that may be used to effect this conversion can be found in the literature, for example j.j.baldwin et al wo 2006042150.

Deprotection of a compound of formula 35 to produce a compound of formula 36 can be achieved by one of ordinary skill in the art using reaction conditions that depend on the nature of the protecting group X. For example, the reaction can be carried out by treating a compound of formula 35 (wherein X represents a t-butoxycarbonyl group) with a protic acid such as trifluoroacetic acid or hydrochloric acid in a suitable inert solvent such as dichloromethane or an ether solvent such as diethyl ether or the like at a temperature of from 0 ℃ to room temperature, preferably at room temperature. Examples of specific conditions for such reactions can be found in t.greene and p.wuts, Protective Groups in Organic Synthesis,3rd.ed., john wiley and Sons (1999).

The compound of formula 36 can be conveniently reduced to the corresponding amine of formula 9 using methods well known to those skilled in the art of organic synthesis. For example, reduction of a compound of formula 36 with a suitable reducing agent (such as lithium aluminum hydride or lithium borohydride, preferably lithium aluminum hydride, and the like) and in a suitable ethereal solvent (such as tetrahydrofuran and the like) at a temperature between about 0 ℃ and about room temperature (preferably at room temperature) affords a compound of formula 9.

Alternative preparation of certain aldehydes of formula 11

Scheme 8

In which R is₂Where hydrogen is indicated, an additional scheme for the preparation of compounds of formula 11 is shown in scheme 8. According to this method, 3- (bromomethyl) benzonitrile (i.e., the compound of formula 37) can be reduced to give a compound of formula 83. This intermediate can then be reacted with a sultam of formula 4 to give a compound of formula 11.

3- (bromomethyl) benzonitrile (i.e., the compound of formula 37) is commercially available from commercial vendors such as Aldrich chemical company, Inc., Milwaukee, Wis., USA; alfa Aesar, Ward Hill, MA, USA; TCIAmerica, Portland, OR, USA; and Acros Organics USA, Morris Plains, NJ, USA.

Reduction of a compound of formula 37 to an aldehyde of formula 38 can be conveniently achieved by treating a compound of formula 37 with a reducing agent known in the art to be effective in reducing nitriles to aldehydes in the presence of benzyl halides. An example of such a reducing agent is diisobutylaluminum hydride. The reaction may be carried out by dissolving the compound of formula 37 in an inert solvent such as chlorobenzene at a temperature of about 0 ℃ and adding a solution of diisobutylaluminum hydride in an inert solvent such as toluene or hexane. Specific conditions that can be used to carry out this reaction can be found in the literature, for example, B.C. Bookser and T.C. Bruice J.am.chem.Soc.1991,113, 4208-4218.

The reaction of the compound of formula 4 with the compound of formula 38 to produce the compound of formula 11 can be carried out using a variety of conditions well known to those of ordinary skill in the art. For example, a compound of formula 4 may be reacted with a compound of formula 38 in the presence of a suitable base, such as a metal carbonate (such as cesium carbonate, potassium carbonate, sodium carbonate or lithium carbonate, preferably cesium carbonate), in a suitable solvent, such as N, N-dimethylformamide and/or tetrahydrofuran. The reaction may be carried out at between about 0 ℃ and room temperature (preferably at about room temperature).

Availability of the aldehyde reagent of formula 13

A variety of aldehyde reagents of formula 13 are commercially available from a variety of suppliers, including the following:

·Acros Organics USA,500 American Road,Morris Plains,NJ 07950,USA，

·Aldrich Chemical Company,Inc.,1001 West Saint Paul Avenue,Milwaukee,WI 53233,USA，

·Alfa Aesar,26 Parkridge Road,Ward Hill,MA 01835,USA，。

·Apollo Scientific Ltd.,Whitefield Road,Bredbury,Stockport,Cheshire SK6 2QR,UK，

·Matrix Scientific,P.O.Box 25067,Columbia,SC 29224-5067,USA，

·Oakwood Products,Inc.,1741 Old Dunbar Road,West Columbia,SC29172,USA，

·TCI America,9211 N.Harborgate Street,Portland,OR97203,USA。

in addition to commercially available reagents, the compound of formula 13 can be prepared using a variety of methods well known in the art of organic synthesis. A list of many of these methods can be found in Comprehensive organic transformations A Guide to Functional Group Preparations R.C.Larock, VCHPublishers, Inc.New York,1989, for example at page 604-. Some of the most common reactions used to prepare aldehydes of formula 13 include oxidation of benzyl alcohol (e.g., using manganese dioxide, using Swern conditions, using dess-martin iodophor, or using o-iodoxybenzoic acid); reduction of carboxylic acid derivatives (e.g., esters or nitriles) using diisobutylaluminum hydride, sodium dihydrobis (2-methoxyethoxy) aluminate (Red-Al, sodium bis (2-methoxyethoxy) -aluminum hydride), or the like; palladium-catalyzed carbonylation; lithium-halogen exchange followed by reaction of the anion with a formamide (such as N-formylpiperidine or N, N-dimethylformamide); or by oxidative cleavage of the double bond of the vinyl-benzene derivative.

Examples of precise conditions suitable for carrying out the oxidation of benzyl alcohol to benzaldehyde can be found in the literature, for example, j.s.yadav et al.tetrahedron 2004,60, 2131-; c.kuhakam et al. Commun.2006,36, 2887-2892; theeraldanon et al tetrahedron 2004,60, 3017-3035; zhao and a. thurkauf Synth. Commun.2001,31, 1921-1926; A.W.white et al.J.Med.chem.2000,43, 4084-; J.Clayden et al tetrahedron 2004,60, 4399-4412; N.Maezaki et al.tetrahedron 2000,56, 7927-; A.P Combs et al.J.Med.chem.2006,49, 3774-3789; and R.M.Moriarty et al.J.org.chem.2004,68, 1890-.

Examples of precise conditions suitable for carrying out the reduction of a carboxylate to benzaldehyde can be found in the literature, for example, n.nakane et al.j.org.chem.2004,69, 3538-3545; T.Abe et al.tetrahedron 2001,57, 2701-2710; and R.Kanazawa and T.Tokoroyama Synthesis 1976, 526-.

Examples of suitable precise conditions for carrying out the reduction of a nitrile to benzaldehyde can be found in the literature, for example, d.castellnou et al.tetrahedron 2005,61, 12111-; T.Itoh et al.J.am.chem.Soc.2006,128, 957-967; E.David et al.J.org.chem.2005,70, 3569-3573; and b.d.roth et al.j.med.chem.1990,33, 21-31.

Examples of the precise conditions suitable for carrying out the conversion of bromo-or iodo-benzene derivatives to benzaldehyde (by metal-halogen exchange followed by formylation) may be found in the literature, e.g. T.and J.Serwatoxski Tetrahedron Lett.2007,48, 1169-; c.g. oliveri et al.j.am.chem.soc.2006,128, 16286-16296; S.Fergus et al.J.org.chem.2004,69, 4663-4669; and S.Hibino et al.heterocycles 1989,28, 275-282.

Examples of precise conditions suitable for carrying out palladium-catalyzed carbonylation of halobenzene derivatives and the like can be found in the literature, for example, K.Orito et al.J.org.chem.1999,64, 6583-6596; R.W.Bates et al.tetrahedron 1995,51, 8199-9212; and H.Iwamoto et al tetrahedron Lett.2002,43, 8191-.

Examples of precise conditions suitable for carrying out oxidative cleavage of the double bond of vinyl-benzene derivatives can be found in the literature, for example, a.srikrishna and g.satyanarayana Tetrahedron 2006,62, 2893-2900; H.Maeda et al.J.org.chem.2005,70, 9693-9701; hashimoto et al bioorg.med.chem.2005,13, 3627-; s.lai and d.g.lee synthesis2001, 1645-1648; y. z.hu and d.l.j.global j.chem.soc.perkin trans.i1997, 1421-1424; S.Rao Kasibhatla et al.J.Med.chem.2000,43, 1508-; and D.Yang and C.Zhang J.org.chem.2001,66, 4814-.

Various aldehydes of formula 13 can also be prepared from benzene derivatives having acidic protons (such as, for example, benzene derivatives containing a hydrogen on the carbon adjacent to the carbon carrying the fluoro group). Such benzene derivatives may be treated with a strong amide base, such as lithium diisopropylamide, in an inert solvent, such as tetrahydrofuran, at low temperatures, such as between about-78 ℃ and about-50 ℃, followed by the addition of a formyl source, such as N, N-dimethylformamide or N-formyl-piperidine, also at a temperature of about-78 ℃. Examples of precise conditions suitable for carrying out this reaction can be found in the literature, for example, A.J.bridges et al.tetrahedron Lett.1992,33, 7499-7502; A.J. Cantrell et al.J. Med.chem.1996,39, 4261-; T.Akama.J.Med.chem.1998,41, 2056-2067; and R.J.Mattson et al.J.org.chem.1999,64, 8004-.

Preparation of an alternative carboxylic acid of formula 29

A plurality of carboxylic acids of formula 29 or lower alkyl esters thereof (wherein R is₅As alkyl, aryl or hydrogen) are commercially available from a variety of vendors including the following:

·Acros Organics USA,Morris Plains,NJ 07950,USA，

·Aldrich Chemical Company,Inc.,Milwaukee,WI 53233,USA，

·Alfa Aesar,Ward Hill,MA 01835,USA，

·Apollo Scientific Ltd.,Stockport,Cheshire SK6 2QR,UK，

·Bachem California Inc.,Torrance,CA,USA，

·Chem-Impex International,Inc.,Wood Dale,IL 60191,USA，

·Oakwood Products,Inc.,West Columbia,SC 29172,USA，

·Sigma-Aldrich Corporation,St.Louis,MO,USA，

·TCI America,Portland，OR 97203,USA，

·3B Scientific Corporation,Libertyville,IL 60048,USA。

in cases where compounds of formula 29 are not commercially available, they can be prepared using methods well known in the art of amino acid chemistry and peptide chemistry. Various reviews of the synthesis of amino acids have been published, including the following: J. A.Ma Angew.chem.Intl.Edn.Engl.2003,42, 4290-; barrett Amino Acids, pept prot.2001,32, 1-106; f.a. davisand b. -c.chen chem.soc.rev.1998,27, 13-18; n.j.turner curr.org.chem.1997,1, 21-36; m.j.burk et al.pure appl.chem.1996,68, 37-44; y.n. belokon Pure appl.chem.1992,64, 1917-1924; and H.E.Shoemaker et al.Pure appl.chem.1992,64, 1171-. These reviews describe a number of different methods that can be used to synthesize compounds of formula 29.

Of the formula HNR₃R₄Availability of amine reagent(s)

Multiple formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl) are commercially available and examples thereof are shown below.

The following are commercially available HNRs of the formula₃R₄Compound of (2) (wherein R is₃Is hydrogen and R₄Is cycloalkyl optionally substituted with alkyl).

Purchased from Sigma-Aldrich Corporation, Saint Louis, MO 63178, USA: (-) -isopinocampheylamine; (+) -isopinocampheylamine; (R) - (+) -bornylamine; adamantan-1-amine; 2-methylcyclohexylamine; 4-methylcyclohexylamine; a cyclobutyl amine; cycloheptylamine; cyclohexylamine; cyclooctylamine; a cyclopentylamine; cyclopropylamine; exo-2-aminonorbornane.

Purchased from Alfa-Aesar, Ward Hill, MA,01835, USA: 1-amino-2-methylcyclohexane; 4-tert-butylcyclohexylamine.

Available from Allichem LLC, Savage, MD, 20763-: 3,3,5, 5-tetramethyl-cyclohexylamine; 4, 4-dimethylcyclohexylamine; trans-2-methylcyclohexylamine.

Purchased from APAC Pharmaceutical, LLC, Columbia, MD,21045, USA: trans-2-methylcyclohexylamine.

Available from Aurora Fine Chemicals LLC, San Diego, CA,92126, USA: 2-ethylcyclopentylamine; 2-methylcyclopentylamine; hexahydro-2, 5-methano-pentalen-3 a (1H) -amine.

Available from 3B Scientific Corporation, Libertyville, IL60048, USA: (±) -2-endo-amino-3-exo-isopropylbicyclo [2.2.1] heptane hydrochloride; 2, 6-dimethylcyclohexylamine; cis-1-amino-4-tert-butylcyclohexane; l-menthyl amine.

Available from chemcridge Corporation, San Diego, CA 92127, USA: (1-isopropylcyclopropyl) amine; (3-ethyl-1-adamantyl) amine; (3-isopropyl-1-adamantyl) amine; bicyclo [2.2.1] heptan-2-amine.

Purchased from Chemgenx LLC, Rocky Hill, NJ,08553, USA: cis-4- (1-methylethyl) -cyclohexylamine.

Purchased from Enamine, Kiev 01103, Ukraine: 2-ethylcyclohexylamine; 2-isopropyl-5-methylcyclohexylamine; decahydronaphthalen-2-amine.

Purchased from Matrix Scientific, Columbia, SC 29224-: 1-methyl-cyclobutylamine; 2-tert-butylcyclohexylamine; 3,5, 7-trimethyl-adamantan-1-ylamine; 4- (1, 1-dimethylpropyl) cyclohexylamine; 4-ethylcyclohexylamine.

Purchased from SYNCHEM OHG, Felsberg-Altenburg, D-34587, Germany: 1-amino-3-methyladamantane; 2-amino-2-methyladamantane.

Purchased from TCI America, Portland, OR 97203, USA: 3,3, 5-trimethylcyclohexylamine; trans-4-methylcyclohexylamine.

Available from TimTec LLC, Newark, DE 19711, USA: 2-aminoamantadine; 2-tert-butyl-cyclohexylamine; memantine.

Purchased from ukrrogsynthesis, Kiev,01133, Ukraine: 1-amino-1-methylcyclohexane; 4-propylcyclohex-1-amine.

Except for the commercially available HNR of formula₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl), various methods for preparing such amines are well known to those of ordinary skill in the art of organic synthesis. Many of these methods are listed in "The Chemistry of The AminoGroup" [ m.s.gibson; s. Patai Ed.; john Wiley& Sons,Ltd.London 1968,37-77]，“Advanced Organic Chemistry”[J.March,3^rd Edition,John Wiley &Sons,Inc.New York,1985]Page 1153-1154 and "Comprehensive organic transformations: A Guide to Functional Group Preparations" [ R.C.Larock, VCHPublishers, Inc.New York,1989]Page 1061-1063. As formula HNR₃R₄An example of the preparation of an amine of (a) can be obtained by treating a ketone with hydroxylamine hydrochloride in an inert solvent such as ethanol at about the reflux temperature of the solvent to convert the ketone such as (1R) - (+) -camphor to the corresponding oxime. The corresponding oxime may then be dissolved in an alcohol (such as amyl alcohol) and then treated with sodium added in small pieces over a longer period of time (such as about four hours) at a temperature of about the reflux temperature of the solvent, resulting in an aminic HNR₃R₄Wherein in this case is (-) -endo-bornylamine hydrochloride, is wherein R₃Represents hydrogen and R₄Compounds representing a bornyl moiety. The precise conditions for carrying out this reaction can be found in the literature, for example, L.A. Patette and R.F.Doehner, Jr.J.org.chem.1980,45,5105 and 5113.

Of the formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl) can be prepared from cyclic ketones by: the ketone is treated with hydrogen and ammonia in the presence of a noble metal catalyst (such as palladium or ruthenium, each of which may optionally be supported on carbon) in the presence of an optional additional ammonium chloride at a temperature of about 200 ℃. The precise conditions for such reactions can be found in the literature, for example, T.Ikenaga et al tetrahedron 2005,61, 2105-.

Of the formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl which is optionally substituted with alkyl) can be prepared from cycloolefins by: the cycloalkene is treated with borane-tetrahydrofuran complex in an inert solvent (such as tetrahydrofuran) at about room temperature to form the corresponding organoborane, which is then treated with chloramine in the presence of aqueous sodium hydroxide. Alternatively, organoboranes can be treated with a solution of hydroxylamine-O-sulfonic acid in diglyme at about 100 ℃ to provide HNRs of the formula₃R₄The amine of (1). The precise conditions for such reactions can be found in the literature, for example, H.C. Brown et al tetrahedron 1987,43, 4071-.

Of the formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl) can be prepared from the formula HOR by₄The alcohol preparation of (1): converting the alcohol to the corresponding formula N₃R₄Followed by reduction of the azide.

Substituted HOR₄The hydroxy group of the alcohol to give the corresponding azido analog can be prepared by using 1, 8-diazabicyclo [ 5.4.0%]Treatment of undec-7-ene (DBU) under anhydrous conditions at a temperature between about 0 ℃ and about 10 ℃ in an inert solvent such as toluene or N, N-dimethylformamide₄And diphenylphosphoryl azide (DPPA) for about 18 hours. Essence for carrying out such a reactionCertain conditions may be found in the literature, for example, P.Bremond et al. Synthesis 2009, 290-; P.Wyrebek et al.tetrahedron 2009,65, 1268-; h.ryu et al.j.med.chem.2008,51, 57-67; or I.Izquierdo et al tetrahedron 2006,63, 1440-1447.

Hydrogenation of the above azido derivatives to the corresponding HNR of formula₃R₄Amine (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl) can be performed in the presence of 5% Pd/C under a hydrogen pressure of between about atmospheric pressure and about 350psi in an organic solvent (such as ethyl acetate, methanol, or ethanol) at room temperature for 1.5 hours. The precise conditions for carrying out such reactions can be found in the literature, for example, m.enomoto and s.kuwarahara angela angelw.chem.intl.edn.engl.2009, 48, 1144-; t.oii et.us 2009131716; X.Wang et al.tetrahedron 2007,63, 6141-6145; or N.Ciliberti et al.Bioorg.Med.chem.2007,15, 3065-.

Alternatively, reduction of the azide group affords a compound of formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl) can be achieved by treating the azide with triphenylphosphine in an inert solvent, such as tetrahydrofuran, in the presence of water at a temperature between about room temperature and about 65 ℃. The precise conditions for carrying out such reactions can be found in the literature, for example, b.han et al.wo 2008148689; liu et al org.Lett.2009,11, 1143-1146; X.Wang et al.tetrahedron 2007,63, 6141-6145; or I.Shimada et al.Bioorg.Med.chem.2008,16, 1966-.

Of the formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Cycloalkyl optionally substituted with alkyl) can be prepared from HOR with hydrocyanic acid using the Ritter reaction₄The tertiary alcohol of (4). According to the process, concentrated sulfuric acid is added to the formula HOR₄And potassium cyanide in dibutyl ether and heating the mixture at about 40 ℃ to obtain a compound of the formula HNR₃R₄The amine of (1). The precise conditions for carrying out such reactions may be found in the literature, for exampleM.Mousseron et al.Bull.Soc.Chim.France 1957,596-600 (U.S. chemical abstracts 51: 76818).

Multiple formula HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Benzyl optionally substituted with halogen) are commercially available and examples thereof are shown below. .

Purchased from Acros Organics, gel, Belgium: 4-bromo-2-fluorobenzylamine.

Purchased from Sigma-Aldrich Corporation, Saint Louis, MO 63178, USA: 2, 3-dichlorobenzylamine; 2, 3-difluorobenzylamine; 2, 4-dichlorobenzylamine; 2, 4-difluorobenzylamine; 2, 5-dichlorobenzylamine; 2, 5-difluorobenzylamine; 2, 6-difluorobenzylamine; 2-chlorobenzylamine; 2-fluorobenzylamine; 3, 4-dichlorobenzylamine; 3, 4-difluorobenzylamine; 3, 5-difluorobenzylamine; 3-chlorobenzylamine; 3-fluorobenzylamine; 3-iodobenzylamine; 4-bromobenzylamine; 4-chlorobenzylamine; 4-fluorobenzylamine; 5-bromo-2-fluorobenzylamine hydrochloride; benzylamine.

Purchased from Alfa-Aesar, Ward Hill, MA,01835, USA: 2,3,4, 6-tetrafluorobenzylamine hydrochloride; 2,3, 4-trifluorobenzylamine; 2,3, 5-trifluorobenzylamine; 2,3, 6-trifluorobenzylamine; 2, 3-dichloro-6-fluorobenzylamine; 2,4, 5-trifluorobenzylamine; 2,4, 6-trifluorobenzylamine; 2, 4-dichloro-5-fluorobenzylamine; 2, 6-dichlorobenzylamine; 2-bromobenzylamine; 2-chloro-3, 6-difluorobenzylamine; 2-chloro-4, 5-difluorobenzylamine; 2-chloro-4-fluorobenzylamine; 2-chloro-5-fluorobenzylamine; 2-chloro-6-fluorobenzylamine; 3,4, 5-trifluorobenzylamine; 3, 5-dichlorobenzylamine; 3-bromo-4-fluorobenzylamine hydrochloride; 3-bromobenzylamine; 3-chloro-2, 4-difluorobenzylamine; 3-chloro-2, 6-difluorobenzylamine; 3-chloro-2-fluorobenzylamine; 3-chloro-4-fluorobenzylamine; 3-chloro-5-fluorobenzylamine; 4-chloro-2, 6-difluorobenzylamine; 4-chloro-2-fluorobenzylamine; 4-chloro-3-fluorobenzylamine; 4-iodobenzylamine; 5-chloro-2-fluorobenzylamine.

Available from Allichem LLC, Savage, MD, 20763-: 5-bromo-2- (aminomethyl) -1, 3-difluorobenzene; 5-bromo-2, 3-difluorobenzylamine.

Available from 3B Scientific Corporation, Libertyville, IL60048, USA: 2-bromo-4-fluorobenzylamine hydrochloride; 2-bromo-5-fluorobenzylamine; 2-iodobenzylamine.

Purchased from Beta Pharma, inc., New Haven, CT 06511, USA: 2,3, 5-trichlorobenzylamine; 2,3, 6-trichlorobenzylamine; 3, 5-dibromobenzylamine; 3-bromo-5-fluorobenzylamine hydrochloride.

Purchased from Enamine, Kiev 01103, Ukraine: 1- (5-bromo-2-fluorophenyl) methylamine.

Purchased from Fluorochem ltd., Old Glossop, deryshire SK 137 RY, united kingdom: 5-chloro-2, 4-difluorobenzylamine.

Except for the commercially available HNR of formula₃R₄Amines of (wherein R is₃Is hydrogen and R₄Benzyl optionally substituted with halogen), various methods for preparing such amines are well known to those of ordinary skill in the art of organic synthesis. Many of these methods are listed in "The Chemistry of The AminoGroup" [ m.s.gibson; s. Patai Ed.; john Wiley& Sons,Ltd.London 1968,37-77]，“Advanced Organic Chemistry”[J.March,3^rd Edition,John Wiley &Sons,Inc.New York,1985]Page 1153-1154 and "Comprehensive organic transformations: A Guide to Functional Group Preparations" [ R.C.Larock, VCHPublishers, Inc.New York,1989]Page 1061-1063.

Can be used for preparing HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄An example of a process for the benzyl group, optionally substituted with halogen, provided that the halogen is stable to the reaction conditions, is the catalytic hydrogenation of benzonitrile. According to this process, a nitrile of formula ArCN (wherein the aryl group Ar represents a benzyl group R) is treated with hydrogen in the presence of a noble metal catalyst (such as palladium, nickel or cobalt) in an inert solvent (such as ethanol) at about room temperature₄Aromatic moiety of (a). The precise conditions for carrying out such reactions can be found in the literature, for example, l.hegedus et al.appl.catal.a.2005,296, 209-215; or F.E.Gould et al.J.org.chem.1960,25, 1658-1660.

Alternatively, a nitrile of formula ArCN (wherein the aryl group Ar represents a benzyl group R)₄Aromatic moiety) can be carried out under elevated hydrogen pressure, such as at about 50 bar, in the presence of a homogeneous catalyst, such as a mixture of bis (2-methallyl) -1, 5-cyclooctadieneruthenium (II), 1-bis (diphenylphosphino) ferrocene, and potassium tert-butoxide in toluene, at about 80 ℃ using conditions similar to those disclosed in s.enthale et al.chem.eur.j.2008,14, 9491-9494.

As a further alternative, a nitrile of formula ArCN (wherein the aryl group Ar represents a benzyl group R)₄Aromatic moiety) can be carried out by treating the nitrile with diisopropylamino borane in the presence of a catalytic amount of lithium borohydride in an inert solvent, such as tetrahydrofuran, at a temperature of about room temperature using conditions similar to those disclosed in d.haddenham et al.j.org.chem.2009,74, 1964-.

Can be used for preparing HNR₃R₄Amines of (wherein R is₃Is hydrogen and R₄Benzyl optionally substituted with halogen) is the conversion of a benzyl halide to benzyl azide followed by reduction of the azide to yield benzylamine. According to the process, the formula R₄The benzyl halide of X, where X represents a leaving group such as a halide (e.g., bromine, chlorine, iodine), an alkyl or aryl sulfonate (e.g., mesylate or tosylate), is reacted with an alkali metal azide salt such as sodium azide in an inert solvent such as dimethyl sulfoxide or ethanol at between about room temperature and about 80 deg.C₃R₄Amine (wherein R is₃Is hydrogen and R₄Is cycloalkyl) reduction of the group of the resulting azide under conditions such as those described for the preparation of the azide.

Multiple formula HNR₃R₄Amines of (a)In R₃And R₄Piperidinyl groups substituted with one or more substituents independently selected from hydroxy and phenyl optionally substituted with halogen, together with the nitrogen atom to which they are attached) are commercially available and examples thereof are shown below.

Purchased from Sigma-Aldrich Corporation, Saint Louis, MO 63178, USA: piperidine; 4-hydroxypiperidine; 4- (4-chlorophenyl) -4-hydroxypiperidine; 3-hydroxypiperidine; 4-phenylpiperidine; 4- (4-bromophenyl) piperidin-4-ol; 4-hydroxy-4-phenylpiperidine; (R) -3-hydroxypiperidine hydrochloride; (S) -3-hydroxypiperidine hydrochloride.

Purchased from Matrix Scientific, Columbia, SC 29224-: 2-phenylpiperidine; 3-phenylpiperidine; 4- (3-fluorophenyl) -piperidine hydrochloride; 4- (4-fluoro-phenyl) -piperidine hydrochloride; 4- (4-fluorophenyl) -4-hydroxypiperidine; 4, 4-diphenylpiperidine hydrochloride; 3-phenylpiperidine hydrochloride; 2-phenylpiperidine hydrochloride; 4- (2-fluorophenyl) piperidine hydrochloride; 3- (4-fluorophenyl) piperidine hydrochloride; 3- (3-fluorophenyl) piperidine hydrochloride; 3- (2-fluorophenyl) piperidine hydrochloride; 2- (4-fluorophenyl) piperidine; 2- (2-fluorophenyl) piperidine; 4- (2-fluoro-phenyl) -piperidin-4-ol.

Purchased from Oakwood Products, inc., West Columbia, SC, USA: 4- (4-chloro-phenyl) -piperidine hydrochloride; 4- (3-chlorophenyl) piperidine hydrochloride; 4- (4' -bromophenyl) piperidine; 4- (3-fluoro-phenyl) -piperidine; 4- (3, 5-dichloro-phenyl) -piperidine; 4- (2, 4-difluorophenyl) piperidine; (S) -3-phenylpiperidine; 4- (4-fluoro-phenyl) -piperidin-4-ol hydrochloride; 4- (3, 5-difluorophenyl) piperidine hydrochloride; 3-hydroxy-3-phenyl-piperidine; 4- (4-bromo-phenyl) -piperidine hydrochloride; 4- (3-bromo-phenyl) -piperidine hydrochloride.

Purchased from ukrrogsynthesis, Kiev,01133, Ukraine: 4- (4-fluorophenyl) piperidine; 4- (2-fluorophenyl) piperidine; 2- (3-fluorophenyl) piperidine; 4- (3, 5-difluorophenyl) piperidine; 2- (2, 4-difluorophenyl) piperidine; 2- (2, 5-difluorophenyl) piperidine; 4- (3, 4-difluorophenyl) piperidine; 2- (2, 6-difluorophenyl) piperidine; 2- (3, 4-difluorophenyl) piperidine; 4- (2, 6-difluorophenyl) piperidine; 4- (2, 5-difluorophenyl) piperidine.

Purchased from Chem-imprex International, inc., Wood Dale, IL 60191, USA: 2- (4-chlorophenyl) piperidine hydrochloride; 4- (4-chlorophenyl) -4-phenylpiperidine hydrochloride; 3, 3-diphenylpiperidine hydrochloride; 4-phenylpiperidine hydrochloride; 2- (4-fluorophenyl) piperidine hydrochloride; 2- (4-bromo-phenyl) -piperidine; 4- (4-fluorophenyl) -4-phenylpiperidine hydrochloride.

Available from 3B Scientific Corporation, Libertyville, IL60048, USA: 4- (4-chlorophenyl) piperidine; 4- (2-chloro-phenyl) -piperidine hydrochloride; 4- (3, 4-difluoro-phenyl) -piperidine hydrochloride; 1, 5-dideoxy-1, 5-imino-xylitol.

Available from Allichem LLC, Savage, MD, 20763-: 4- (3-bromo-phenyl) -piperidine; 4- (3-chlorophenyl) piperidine.

Purchased from Beta Pharma, inc., New Haven, CT 06511, USA: 4, 4-diphenylpiperidine; 4- (4-bromophenyl) -4- (4-chlorophenyl) piperidine; 4, 4-bis (4-chlorophenyl) piperidine.

Available from chemcridge Corporation, San Diego, CA 92127, USA: 3- (4-chlorophenyl) piperidine.

In addition to the commercially available HNR of formula₃R₄Compound of (2) (wherein R is₃And R₄Piperidinyl which together with the nitrogen atom to which it is attached forms a phenyl group substituted at the 4-position with a hydroxy group and optionally with halogen), further compounds of this type can be prepared using reactions well known in the art of organic synthesis.

For example, such compounds can be prepared by the following method: treatment of aryl bromide with magnesium metal in ether at about room temperature gives the corresponding Grignard reagent or with a solution of n-butyllithium in tetrahydrofuran at about-78 ℃ gives the corresponding organolithium reagent, which can then be treated with either of the above reagents in the same solvent at between about-78 ℃ and about room temperature to give the alcohol of formula 39. The precise conditions for carrying out such reactions can be found in the literature, for example, m.tomishima et al.bioorg.med.chem.lett.2008,18, 2886-; sonesson et al us 20080269286; g. chang et al.us 20070213371; or g.chiu et al.us 20060217419. The t-butoxycarbonyl protecting group may then be removed in the usual manner, for example by treatment with a solution of HCl in dioxane or trifluoroacetic acid in dichloromethane at about room temperature to give the desired piperidine. The precise conditions for carrying out such reactions can be found in the literature, for example, p.zhang et al.us 20070088036; Y.Jiang et al.J.Med.chem.2007,50, 3870-3882; C.Zhi et al.J. Med.chem.2005,48, 7063-; and G.Chiu et al.Bioorg.Med.chem.Lett.2007,17, 3930-.

Of the formula HNR₃R₄Compound of (2) (wherein R is₃And R₄Piperidinyl groups which together with the nitrogen atom to which they are attached form a phenyl group substituted at the 4-position with optionally substituted halogen) can be prepared using reactions well known in the art of organic synthesis. For example, such compounds can be prepared from compounds of formula 39 by sequential dehydration, reduction, and removal of the t-butyloxycarbonyl protecting group. The dehydration reaction can be achieved by treating the compound of formula 39 with a solution of methanesulfonyl chloride and triethylamine in dichloromethane at a temperature of about-78 ℃ to provide the compound of formula 40. Hydrogenation of the compound of formula 40 can be achieved by treating the resulting olefin with hydrogen in the presence of a noble metal catalyst (such as Pd/C) in an inert solvent (such as ethanol or ethyl acetate) at a hydrogen pressure between about 14psi and about 50psi at about room temperature. Finally, removal of the t-butoxycarbonyl protecting group may be accomplished as described in the preceding paragraph. The precise conditions for carrying out such a reaction sequence can be found in the literature, for example, G.Chiu et al.Bioorg.Med.chem.Lett.2007,17, 3930-.

The compound of formula 40 can also be prepared by converting 1-tert-butoxycarbonylpiperidin-4-one to the triflate derivative of formula 41. The reaction may conveniently be carried out as described in Receveur, j. -m.et al US20090062317, wherein 1-tert-butoxycarbonylpiperidin-4-one is added to a solution of lithium diisopropylamide in tetrahydrofuran at-78 ℃ for 30 minutes, N-phenyl-trifluoromethanesulfonimide is added and stirred overnight. The resulting compound of formula 41 can then be reacted with one of a variety of phenyl boronic acids to provide a compound of formula 40. This reaction, known to those of ordinary skill in the art of organic synthesis as the Suzuki reaction, can be carried out by treating the triflate derivative of formula 41 with phenylboronic acid in the presence of a catalyst such as tetrakis (triphenylphosphine) palladium (0) or bis (triphenylphosphine) palladium (II) dichloride, a base such as potassium carbonate, in an inert solvent such as dimethoxyethane or dioxane at a temperature of about 80 ℃. The precise conditions for carrying out such reactions can be found in the literature, for example, Receveur, j. -m.et al US 20090062317; J.M.Goss and S.E.Schaus J.org.chem.2008,73, 7651-7656; or m.r.dobler et al.us 20080255149.

The invention also relates to compositions and unit dose formulations comprising the compounds of the invention. The compositions and unit dose formulations include a therapeutically effective amount of a compound of the invention and a carrier. The compositions and unit dose formulations may also include additional adjuvants, such as other excipients. Generally, from about 1% to about 99%, preferably from about 5% to about 70% and most preferably from about 10% to about 30% of the composition or unit dose formulation is comprised of a compound of the present invention.

Lactose, corn starch or derivatives thereof, talc, stearic acid or its salts and the like can be used, for example, as carriers for tablets, coated tablets, dragees and hard gelatine capsules. Suitable carriers for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols and the like. However, depending on the nature of the active substance, no carriers are generally required in the case of soft gelatin capsules. Suitable carriers for the production of solutions and syrups are, for example, water, polyols, glycerol, vegetable oils and the like. Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

The compositions and unit dose formulations of the present invention may also include additional excipients such as preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavors, salts for varying osmotic pressure, buffers, masking agents, and antioxidants.

The compositions and unit dose formulations of the present invention may also include additional therapeutically active agents.

Unit dose formulations of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The formulations may be prepared by any method known in the pharmaceutical art.

Unit dose formulations of the present invention suitable for oral administration may be in the form of capsules, cachets, sachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, elixirs, syrups, pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia), mouth washes and the like. The formulations may also be solutions or suspensions of the compounds of the invention in aqueous or non-aqueous liquids. The formulation may also be an oil-in-water or water-in-oil liquid emulsion. The compounds of the invention may also be administered in the form of a bolus, electuary or paste.

The invention also relates to methods for preparing the compositions and unit dose formulations of the invention. Such methods include the step of admixing a compound of the present invention with a carrier and optionally one or more adjuvants. Generally, the compositions and formulations of the present invention are prepared by uniformly and intimately bringing into association a compound of the invention with liquid carriers, finely divided solid carriers or both, and then, if necessary, shaping the product.

The invention also relates to a method for treating a patient suffering from a proliferative disorder, said method comprising the step of administering to said patient a compound of the invention. The compounds may be contained in a composition or unit dose formulation. In a preferred embodiment, the proliferative disorder is a solid tumor. In a particularly preferred embodiment, the proliferative disorder is selected from the group consisting of: breast, lung, colon and prostate tumors.

The following examples and references are provided to aid the understanding of the present invention, the true scope of which is defined in the appended claims.

Examples

Reagents were purchased from Aldrich, Sigma, Maybridge, Advanced ChemTech, and Lancaster or other suppliers indicated below and used without further purification. LC/MS (liquid chromatography/mass spectrometry) spectra were recorded using the following system. For measuring mass spectra, the system was measured by a MicromassPlatform II spectrometer: ES ionization (positive mode) (mass range: 150-1200 amu). The simultaneous chromatographic separation was achieved using the following HPLC system: ES industries Chromegabond WR C-183u(3.2X 30mm) column; mobile phase A: water (0.02% TFA) and phase B: acetonitrile (0.02% TFA); gradient 10% B to 90% B, 3-7 min; an equilibration time of 1 minute; the flow rate was 2 mL/min.

Supercritical fluid chromatography separation was performed using a Mettler-Toledo Minigram system with the following typical conditions: 100 bar, 30 ℃ with 40% in supercritical fluid CO₂The methanol in (1) eluted the 12mm AD column at 2.0 mL/min. In the case where the analyte has a basic amino group, 0.2% isopropylamine is added to the methanol modifier.

Preparation of preferred synthetic intermediates

Intermediate 1:1, 1-dioxo-isothiazolidine-3-carboxylic acid methyl ester

Step 1: (S) -2-amino-4- ((S) -3-amino-3-methoxycarbonyl-propyldithio) -butyric acid methyl ester. Thionyl chloride (0.5mL, 8.20mmol) was slowly added to a stirred solution of (S) -2-amino-4- ((S) -3-amino-3-carboxy-propyldisulfide) -butyric acid (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 1.0g, 3.73mmol) in methanol (12.5mL) at 0 ℃. The mixture was then refluxed for 3h, during which time the reaction was complete (monitored by silica gel TLC). The reaction mixture was cooled to room temperature and the solvent was distilled off under reduced pressure to give crude (S) -methyl 2-amino-4- ((S) -3-amino-3-methoxycarbonyl-propyldithio) -butyrate (1.0g, 90%) as a white solid which was used continuously for the next step without further purification.

Step 2:1, 1-dioxo-1. lamda⁶3-Carboxylic acid methyl ester (isothiazolidine)

A stream of chlorine was bubbled through a cooled solution of (S) -2-amino-4- ((S) -3-amino-3-methoxycarbonyl-propyldithio) -butyric acid methyl ester (0.50g, 1.68mmol) in ethanol/chloroform (1:2,15mL) at 0 ℃ for 1 h. The solvent was distilled off under reduced pressure and the residue was left under high vacuum for 4 h. The residue was then diluted with chloroform (5mL), cooled to-5 ℃ and triethylamine (1.4mL, 9.88mmol) was added dropwise at the same temperature. The reaction temperature was returned to room temperature and the mixture was stirred for 1 h. The solvent was distilled off under vacuum and the crude product was purified by column chromatography on silica gel (100-200 mesh) using a gradient of 1-2% methanol/dichloromethane to give pure 1, 1-dioxo-1. lambda⁶Methyl (0.250g, 41%) isothiazolidine-3-carboxylate.

Intermediate 2, 2-benzyl-1, 1-dioxo-1-lambda⁶3-carboxylic acid (isothiazolidine)

Step 1: to 1, 1-dioxo-1. lamda. at 0 ℃⁶To a stirred solution of methyl (intermediate 1, 0.200g, 1.1mmol) of (i) -isothiazolidine-3-carboxylate in N, N-dimethylformamide (5mL) was added potassium carbonate (0.450g, 3.2 mmol). Stirring at the same temperature for 10min, addingBenzyl chloride (0.153g, 1.2mmol) and tetra-n-butylammonium iodide (0.40g, 1.1mmol) were added. After stirring at room temperature for 5h, the reaction mixture was diluted with ethyl acetate (25mL) and washed with water (3X 15 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude material, which was purified by silica gel column chromatography (100-200 mesh) to give 2-benzyl-1, 1-dioxo-1-. lambda.⁶Methyl isothiazolidine-3-carboxylate (0.170g, 56%). FIA-MS (+1 mode): m/z =270[ M + 1]]。

Step 2: to 2-benzyl-1, 1-dioxo-1-lambda at room temperature⁶To a stirred solution of methyl (0.600g, 2.23mmol) isothiazolidine-3-carboxylate in tetrahydrofuran-water (5:1, 20mL) was added lithium hydroxide monohydrate (0.140g, 3.3 mmol). After stirring at the same temperature overnight, the reaction mixture was acidified with 2N hydrochloric acid (pH. about.3) and extracted with ethyl acetate (3X 25 mL). The combined organic portions were washed with water, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-benzyl-1, 1-dioxo-1-. lambda.⁶Isothiazolidine-3-carboxylic acid (0.420g, 74%) as crude product, which was used continuously for the next step without further purification. FIA-MS (+1 mode): m/z =254[ M + 1]]。

Intermediate 3 (S) -4,4, N1, N1-tetramethyl-pentane-1, 2-diamine hydrochloride

Step 1: (S) -2-amino-4, 4-dimethyl-pentan-1-ol

To a stirred suspension of lithium borohydride (0.75g, 34.4mmol, 2.0 equiv.) in tetrahydrofuran (60mL) was slowly added chlorotrimethylsilane (7.48g, 68.85mmol, 4.0 equiv.) at 0 ℃ under argon over a period of 5 minutes. (S) -2-amino-4, 4-dimethyl-pentanoic acid (purchased from Chem-Impex International, Inc., Wood Dale, IL, USA; 2.50g, 17.2mmol, 1.0 equiv.) is added to the reaction mixture in portions over a period of 10 minutes at the same temperature and the reaction mixture is stirred at room temperature for 24 hours. The reaction mixture was quenched by slow addition of methanol (50mL) at 0 ℃ and volatiles were removed by vacuum distillation. The resulting residue was treated with 20% aqueous potassium hydroxide (w/v; 10mL) and extracted with dichloromethane (3X 20 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give (S) -2-amino-4, 4-dimethyl-pentan-1-ol (1.9g, 84%).

Step 2: ((S) -1-hydroxymethyl-3, 3-dimethyl-butyl) -carbamic acid tert-butyl ester

To a stirred solution of (S) -2-amino-4, 4-dimethyl-pentan-1-ol (2.0g, 15.2mmol, 1.0 equiv.) and triethylamine (1.85g, 18.3mmol, 1.2 equiv.) in anhydrous tetrahydrofuran (50mL) was slowly added di-tert-butyl dicarbonate (3.99g, 18.3mmol, 1.2 equiv.) under nitrogen at 0 ℃. After stirring at room temperature for 4h, the tetrahydrofuran was distilled off under reduced pressure and the crude reaction mixture was purified by silica gel column chromatography (100-200 mesh) using a gradient of 10-25% ethyl acetate/hexane to give ((S) -1-hydroxymethyl-3, 3-dimethyl-butyl) -carbamic acid tert-butyl ester (2.8g, 79%).

And step 3: toluene-4-sulfonic acid (S) -2-tert-Butoxycarbonylamino-4, 4-dimethyl-pentyl ester

To a stirred solution of ((S) -1-hydroxymethyl-3, 3-dimethyl-butyl) -carbamic acid tert-butyl ester (3.0g, 13.0mmol, 1.0 equiv.) in anhydrous pyridine (14.8mL) was added tosyl chloride (3.7g, 19.4mmol, 1.5 equiv.) portionwise at 0 ℃ under nitrogen. After stirring at room temperature for 4 hours, the reaction mixture was extracted with ethyl acetate (200mL), washed with water (3X 200mL) followed by brine (2X 100mL), and dried over anhydrous sodium sulfate to give a crude residue. The residue was purified by silica gel column chromatography (100-200 mesh) using a gradient of 5-10% ethyl acetate/hexane to give toluene-4-sulfonic acid (S) -2-tert-butoxycarbonylamino-4, 4-dimethyl-pentyl ester (3g, 60%).

And 4, step 4: ((S) -1-Dimethylaminomethyl-3, 3-dimethyl-butyl) -carbamic acid tert-butyl ester

Dimethylamine (11.2g, 249.3mmol, 30.0 equiv.) was added portionwise to a stirred solution of toluene-4-sulfonic acid (S) -2-tert-butoxycarbonylamino-4, 4-dimethyl-pentyl ester (3.2g, 8.31mmol, 1.0 equiv.) in pyridine (9.6mL) at 0 ℃. The reaction mixture was slowly warmed to room temperature and stirred at the same temperature for an additional 20 hours. The reaction mixture was diluted with 10% aqueous potassium hydroxide (w/v; 30mL) and extracted with dichloromethane (3X 150 mL). The collected organic extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude material. This material was purified by silica gel column chromatography (100-200 mesh) using a gradient of 2-5% methanol/dichloromethane to give ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -carbamic acid tert-butyl ester (1.4g, 65%).

And 5: (S) -4,4, N1, N1-tetramethyl-pentane-1, 2-diamine hydrochloride

A solution of ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -carbamic acid tert-butyl ester (1.6g, 6.20mmol, 1.0 eq) in anhydrous HCl-ethyl acetate (31mL) was stirred at room temperature under nitrogen overnight. Ethyl acetate was distilled off under reduced pressure to give crude (S) -4,4, N1, N1-tetramethyl-pentane-1, 2-diamine hydrochloride (1.4g) as a white solid.

Intermediate 4: 6-chloro-2-fluoro-3-methoxy-benzaldehyde

A solution of 4-chloro-2-fluoro-1-methoxy-benzene (Aldrich; 10.2g, 63.5mmol) in anhydrous tetrahydrofuran (500mL) was cooled to-78 ℃ under a nitrogen atmosphere. Lithium diisopropylamide solution (1.8M in tetrahydrofuran/heptane/ethylbenzene, 39.9mL, 70.2mmol) was added dropwise via syringe. The reaction mixture was warmed to-55 ℃ and held at this temperature for 1 h. The mixture was then cooled again to-78 ℃ and dry N, N-dimethylformamide (10.7mL, 139mmol) was added via syringe. The cooling bath was removed and the reaction mixture was allowed to warm to-10 ℃ and quenched by the addition of flake ice (ice flake) (about 200mL) and saturated ammonium chloride solution (200 mL). Ethyl acetate (200mL) was added, the layers were separated and the aqueous layer was extracted with ethyl acetate (200 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, evaporated, and purified by silica gel chromatography (eluting with 0-30% ethyl acetate/hexanes) to give 6-chloro-2-fluoro-3-methoxy-benzaldehyde (5.6g, 47% yield) as an oil that solidified upon standing.

Intermediate 52- (3-formyl-benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid ethyl ester

Step 1: 3-bromomethyl-benzaldehyde

A solution of 3-cyanobenzyl bromide (purchased from Aldrich; 10.1g, 51.5mmol) in chlorobenzene (100mL) was cooled in an ice-water bath. A solution of diisobutylaluminum hydride in hexane (from Aldrich; 1M; 65mL, 65mmol) was added over 25 minutes and the reaction mixture was stirred for 1h at 5 ℃ below. Chloroform (100mL) and 10% aqueous hydrochloric acid were added sequentially (dropwise). The layers were separated and the organic layer was washed with water. The aqueous layers were back-extracted with chloroform and the organic phases were combined, dried over sodium sulfate, filtered, evaporated and purified by chromatography (eluting with 0-40% ethyl acetate/hexane) to give pure fractions and various impure fractions. The impure fractions were purified again by chromatography (eluting with 0-27% ethyl acetate/hexane). The homogeneous fractions from the two chromatographic separations corresponding to the product were combined and concentrated. The residue was dissolved in ether and partitioned with hexane. The mixture was frozen overnight and the solid was filtered off, washed with hexane and dried under high vacuum at room temperature to give 3-bromomethyl-benzaldehyde (7.32g, 71%).

Step 2:2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶Isothiazolidine-3-carboxylic acid ethyl ester

3-bromomethyl-benzaldehyde (2.44g, 12.3mmol) and cesium carbonate (9.08g, 27.9mmol) were added to a solution of a 4:3 mixture of methyl 1, 1-dioxo-isothiazolidine-3-carboxylate and ethyl 1, 1-dioxo-isothiazolidine-3-carboxylate (intermediate 7 step 2; 2.01g, 10.8mmol) in N, N-dimethylformamide (60 mL). The mixture was stirred at room temperature for 20h, then dichloromethane (100mL) was added. Saturated aqueous ammonium chloride (25mL) was added followed by 1M hydrochloric acid to bring the pH to 2-3. The two layers were separated and the organic layer was washed twice with brine. The solution was dried over sodium sulfate and filtered. Celite was added and the solvent was evaporated under reduced pressure. The residue (mixture containing crude product coated on celite) was purified by flash chromatography (Analogix SF40-240g Si, eluting with 30-70% ethyl acetate/hexanes) to give 2- (3-formyl-benzyl) -1, 1-dioxo-1 λ⁶Isothiazolidine-3-carboxylic acid ethyl ester (first elution peak; 1.26g, 37%) and 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda. (Lambda.)⁶Methyl isothiazolidine-3-carboxylate (second peak; 1.69g, 53%).

Intermediate 6 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amides

Step 1: ((S) -1-dimethylcarbamoyl-2-phenyl-ethyl) -carbamic acid tert-butyl ester

To a cooled solution of (S) -2-tert-butoxycarbonylamino-3-phenylpropionic acid (from Aldrich; 5.3g, 20mmol) in dimethylformamide (50mL) was added 1-hydroxybenzotriazole (from 3BScientific Corporation, Libertyville, IL60048, USA; 2.97g, 22mmol) and O-benzotriazol-1-yl-N, N, N ', N' -tetramethyluronium hexafluorophosphate (from Aldrich; 8.30g, 22mmol) at 0 ℃. Diisopropylethylamine (10.7mL, 61.4mmol) and dimethylamine in tetrahydrofuran (2M, 15mL, 30mmol) were added dropwise. The mixture was stirred at 0 ℃ for 1h and ethyl acetate was added. The mixture was washed with water and brine, and dried over sodium sulfate. The solvent was evaporated to give ((S) -1-dimethylcarbamoyl-2-phenyl-ethyl) -carbamic acid tert-butyl ester as a yellow oil (5.52g) which was used in the next step without further purification.

Step 2: (S) -2-amino-N, N-dimethyl-3-phenyl-propionamide

((S) -1-dimethylcarbamoyl-2-phenyl-ethyl) -carbamic acid tert-butyl ester (2.0g, 6.85mmol) was dissolved in dichloromethane (20mL) at 0 ℃. Trifluoroacetic acid (20mL) was added. The mixture was stirred for 2h and evaporated. The residue was dissolved in dichloromethane and very carefully basified with saturated aqueous sodium bicarbonate. The organic layer was separated and the aqueous layer was extracted with dichloromethane. The combined organic extracts were washed with water and brine and then dried over sodium sulfate to give (S) -2-amino-N, N-dimethyl-3-phenyl-propionamide (1.0g, 76%) as a yellow oil.

And step 3: (S) -N1, N1-dimethyl-3-phenyl-propane-1, 2-diamine

To a solution of (S) -2-amino-N, N-dimethyl-3-phenyl-propionamide (1.0g, 5.2mmol) in tetrahydrofuran (10mL) was added dropwise a solution of lithium aluminum hydride in tetrahydrofuran and toluene (3.5M, 7.4mL, 26 mmol). After addition the mixture was heated to reflux and held for 4h, then cooled. Ether (20mL) was added. Ethyl acetate was added dropwise until all the lithium aluminum hydride was consumed. Sodium hydroxide (1M, 20mL) was added and the mixture was stirred for 30 min. The mixture was extracted three times with diethyl ether. The combined organic extracts were washed with brine, dried over sodium sulfate and evaporated to give (S) -N1, N1-dimethyl-3-phenyl-propane-1, 2-diamine (0.60g, 65%).

And 4, step 4: 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

To a solution of (S) -N, N' -dimethyl-3-phenyl-propane-1, 2-diamine (120mg, 0.673mmol) in methanol (2mL) under argon was added 2- (3-formyl-benzyl) -1, 1-dioxo-1. lambda⁶A solution of ethyl (intermediate 4; 230.5mg, 0.740mmol) isothiazolidine-3-carboxylate in methanol (4 mL). The solution was cooled to 0 ℃ in an ice bath and to it was added sodium cyanoborohydride (63.4mg, 1.01mmol) and acetic acid (0.3 mL). The solution was allowed to warm to room temperature and stirred under argon for 16 h. The reaction mixture was diluted with ethyl acetate (50mL), washed with water (2X 10mL) and the aqueous layer extracted with ethyl acetate (50 mL). The combined organics were dried over sodium sulfate, filtered and concentrated to give crude 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Ethyl isothiazolidine-3-carboxylate (360mg) as a colorless oil.

And 5:2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶3-carboxylic acid (isothiazolidine)

To crude 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylEthylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶To a solution of ethyl-isothiazolidine-3-carboxylate (360mg, ca. 0.67mmol) in tetrahydrofuran (4mL) was added lithium hydroxide hydrate (57.4mg, 1.37mmol) followed by water (1 mL). It was stirred at room temperature for 2 h. To this stirred solution was added about 15 drops of 1N hydrochloric acid to bring the pH to 4.5. The sample was concentrated to dryness and dried over night with phosphorus pentoxide. Crude 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Isothiazolidine-3-carboxylic acid (350mg, about 80% pure) (in the presence of LiCl) was used in the next step.

Step 6:2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amides

(+) -Isopinocampheylamine (Aldrich; 29mg, 0.19mmol), O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate (from Aldrich; 58.4mg, 0.18mmol), N-hydroxybenzotriazole (from 3B Scientific Corporation, Libertyville, IL60048, USA; 24.3mg, 0.18mmol) was added to 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl ethyl amino) -methyl under argon]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid (100mg 80% pure, 0.18mmol) in a cooled (0 ℃) solution in anhydrous N, N-dimethylformamide (3 mL). N, N-diisopropylethylamine (70. mu.L, 0.4mmol) was then added. The solution was then stirred at room temperature. After 30 minutes, more (+) -isopinocampheylamine (6mg, 0.04mmol), O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate (available from Aldrich; 6mg, 0.002mmol) and N-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 3mg, 0.002mmol) were added. After 90 minutes, all starting material had been consumed. The reaction mixture was diluted with ethyl acetate (50mL) and washed with 1N NaOH (3X 15mL) and brine (20mL)And (6) washing. The organic phase was dried over sodium sulfate, filtered and concentrated to give the crude product (132 mg). This material was mixed with 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Crude products of the same experiment of-isothiazolidine-3-carboxylic acid (250mg) were combined. The combined different batches of product were purified by chromatography (eluting with 0-10% methanol/dichloromethane) to give 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amide (256mg, 60%) as a tan foam.

Intermediate 7 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Step 1: 2- (3-cyano-benzyl) -1, 1-dioxo-1. lamda.⁶3-Carboxylic acid methyl ester (isothiazolidine)

To 1, 1-dioxo-1. lamda. at room temperature under nitrogen⁶To a stirred solution of methyl (intermediate 1, 0.200g, 1.10mmol) of (E) -isothiazolidine-3-carboxylate in N, N-dimethylformamide (5mL) was added potassium carbonate (0.450g, 3.20 mmol). The reaction mixture was cooled to 0 ℃ and 3-cyanobenzyl bromide (from Aldrich; 0.240g, 1.22mmol) was added slowly to the reaction mixture, followed by tetra-n-butylammonium (0.025g, 0.07 mmol). The reaction was completed at rt over 5h (monitored by silica gel TLC). The reaction mixture was diluted with ethyl acetate (25mL), washed with water (3X 15mL), dried over sodium sulfate and concentrated under reduced pressure to give a crude material which was purified by silica gel column chromatography (100-200 mesh) using a gradient of 20-30% ethyl acetate/hexane to give 2- (3-cyano-benzyl) -1, 1-dioxo-1. lambda. hexane⁶Methyl isothiazolidine-3-carboxylate (0.170g, 56%).

Step 2:2- (3-cyano-benzyl) -1, 1-dioxo-1. lamda.⁶3-carboxylic acid (isothiazolidine)

To 2- (3-cyano-benzyl) -1, 1-dioxo-1. lamda. at room temperature⁶To a stirred solution of methyl (6.0g, 20.4mmol) isothiazolidine-3-carboxylate in tetrahydrofuran-water mixture (4:1,50mL) was added lithium hydroxide monohydrate (2.5g, 59.6mmol) and stirred at this temperature for 4 h. The tetrahydrofuran was then removed from the reaction mixture under reduced pressure. The residual reaction mixture was cooled to 0 ℃ and 1N hydrochloric acid was added to bring the pH to about 2. The acidified reaction mixture was extracted with ethyl acetate (3X 150 mL). The combined organic layers were washed with water (2X 50mL) followed by brine (50mL), then dried over sodium sulfate and concentrated under reduced pressure to give 2- (3-cyano-benzyl) -1, 1-dioxo-1. lambda⁶Isothiazolidine-3-carboxylic acid (5.2g, 91%).

And step 3: 2- (3-cyano-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

To 2- (3-cyano-benzyl) -1, 1-dioxo-1. lamda. at room temperature⁶To a stirred solution of isothiazolidine-3-carboxylic acid (6.0g, 21.4mmol) in tetrahydrofuran (40mL) was added simultaneously 1-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 2.8g, 20.72mmol), TBTU (6.8g, 21.18mmol) and diisopropylethylamine (8.3g, 64.21 mmol). After stirring at the same temperature for 16 hours, tetrahydrofuran was distilled off under reduced pressure. The resulting crude residue was extracted with ethyl acetate (120mL), washed with 1N aqueous NaOH (50mL), brine (50mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude material. The crude material was purified by silica gel column chromatography (100-200 mesh) using a gradient of 50-70% ethyl acetate/hexane to give 2- (3-cyano-benzyl) -1, 1-dioxo-1. lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (7.0g, 79.5%).

And 4, step 4: 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]G-channel wine3-yl) -amides

To 2- (3-cyano-benzyl) -1, 1-dioxo-1. lamda. at room temperature⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (0.200g, 0.48mmol) in a mixture of pyridine-acetic acid-water (2:1: 1; 7.2mL) was added sodium hypophosphate hydrate (0.400g, 3.77 mmol). The reaction mixture was cooled to 0 ℃ under an argon atmosphere and raney nickel (0.200g) was added. After stirring at 0 ℃ for 10min, the temperature was raised to 40-45 ℃ and the reaction mixture was stirred at the same temperature for an additional 2.5h (monitored by silica gel TLC). The reaction mixture was cooled, filtered through a pad of celite and the residue was washed with methanol (5 mL). The combined filtrates were concentrated to give a crude material which was purified by silica gel column chromatography (100-200 mesh) using a gradient of 50-60% ethyl acetate/hexane to give 2- (3-formyl-benzyl) -1, 1-dioxo-1. lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (0.100g, 49%).

And 5:2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl ] -benzyl } -

1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

To a stirred solution of (S) -4,4, N1, N1-tetramethyl-pentane-1, 2-diamine hydrochloride (intermediate 2, 0.100g, 0.43mmol) in methanol (5mL) was added triethylamine (0.12g, 0.16mL, 1.2mmol) at 0 ℃. Stirring at the same temperature for 10min, adding 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda. (lambda.)⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (0.181g, 0.43mmol) in methanol (1mL) and stirring continued for an additional 15min at room temperature. The reaction mixture was again brought to 0 ℃ and sodium cyanoborohydride (0.022g, 0.35mmol) and acetic acid (0.2mL) were added simultaneously to the reaction mixture. After stirring at room temperature overnight, the reaction mixture was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (5mL), washed with water (3mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to giveCrude 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (0.216g, 89%).

Intermediate 8 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶3-carboxylic acid (isothiazolidine)

Step 1: (S) -2-amino-4- ((S) -3-amino-3-methoxycarbonyl-propyldithio) -butyric acid methyl ester

To a stirred solution of (S) -2-amino-4- ((S) -3-amino-3-carboxy-propyldithio) -butyric acid (6.0g, 22.3mmol) in methanol (156mL) was slowly added thionyl chloride (6.5mL, 89.5mmol) at 0 ℃. The mixture was then stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give crude (S) -2-amino-4- ((S) -3-amino-3-methoxycarbonyl-propyldithio) -butyric acid methyl ester, which was used continuously in the next step without further purification.

Step 2:1, 1-dioxo-1. lamda⁶3-Thiazolidine-carboxylic acid methyl ester and 1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ethyl ester

To a cooled solution of (S) -2-amino-4- ((S) -3-amino-3-methoxycarbonyl-propyldisulfide) -butyric acid methyl ester (crude material from step 1, 22.3mmol) in ethanol-chloroform (1:2,135mL) was bubbled a stream of chlorine at 0 ℃ for 1 h. The solvent was distilled off under reduced pressure and the crude material was left under high vacuum for 1 h. The resulting residue was diluted with chloroform (30mL) and cooled to 0 ℃. Triethylamine (16mL, 111.8mmol) was added dropwise at the same temperature. The reaction temperature was returned to room temperature and stirred overnight. The solvent was concentrated under vacuum and the crude (1, 1-dioxo-1. lamda.) was purified⁶3-carboxylic acid (isothiazolidine)A mixture of methyl and ethyl esters (6.4g, 80%) (1.3: 1; methyl ester: ethyl ester)) was continued for the next step in the reaction sequence.

And step 3: 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

To 1, 1-dioxo-1. lamda⁶To a stirred solution of a mixture of methyl-and ethyl-isothiazolidine-3-carboxylate (0.085g, 0.45mmol) (1.3: 1; methyl ester: ethyl ester) in dry N, N-dimethylformamide (2.5mL) was added 3-bromomethyl-benzaldehyde (0.100g, 0.50 mmol). The solution was stirred at room temperature under an argon atmosphere for 20 hours. The reaction mixture was diluted with dichloromethane. Saturated ammonium chloride and hydrochloric acid (1.0M) were added sequentially until the mixture reached a pH of about 2-3. The organic layer was separated, washed with brine, dried over anhydrous sodium sulfate and concentrated on celite. The product was purified by silica gel column chromatography to give 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester (0.053g, 42%).

And 4, step 4: 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl }1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ethyl ester

To a stirred solution of (S) -4,4, N1, N1-tetramethyl-pentane-1, 2-diamine hydrochloride (intermediate 2, 0.921g, 4.73mmol) in methanol (10mL) cooled in an ice bath (0 ℃) was added triethylamine (2.19mL, 15.7mmol) and the solution was stirred at this temperature for 25 minutes. To the cooled solution was added 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶-a solution of ethyl isothiazolidine-3-carboxylate (0.937g, 3.15mmol) in methanol (35mL) after which the solution was allowed to warm to room temperature and stirred for 2 hours. The solution was cooled to 0 ℃, sodium cyanoborohydride (0.461g, 7.3mmol) and acetic acid (4mL) were added and the solution was allowed to warm to room temperature and stirred overnight. The reaction mixture was then concentrated under reduced pressure, partitioned between dichloromethane and saturated aqueous sodium bicarbonate and the layers separated. The organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl }1,1-Dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ethyl ester, which is used continuously in the next step without further purification.

And 5:2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ethyl ester

2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl }1, 1-dioxo-1. lamda⁶Ethyl-isothiazolidine-3-carboxylate (1.1g, 2.4mmol) was dissolved in methanol (60mL) and the solution was cooled to 0 ℃ and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 0.750g, 3.98mmol) was added in one portion. The ice bath was removed and the solution was stirred at room temperature for 40 minutes, after which the solution was cooled to 0 ℃. To the cooled solution was added sodium cyanoborohydride (0.400g, 6.37mmol) and acetic acid (0.485mL) and the solution was stirred at room temperature for 5 hours, after which another portion of sodium cyanoborohydride (0.200g, 3.18mmol) was added. The solution was stirred at room temperature overnight, after which it was concentrated under reduced pressure. The reaction mixture was partitioned between dichloromethane and water, rinsed with saturated aqueous sodium bicarbonate solution and the layers separated. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude material which was purified by reverse phase chromatography (Gilson; 50Polaris C18A (50g), 40% -100% acetonitrile/water (0.1% TFA) over 6 minutes) to give 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino after neutralization with sodium bicarbonate to remove trifluoroacetic acid]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ethyl ester (0.87g, 58%).

Step 6:2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶3-carboxylic acid (isothiazolidine)

Coupling 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶3-carboxylic acid (isothiazolidine)Ethyl ester (0.865g, 1.4mmol) was dissolved in tetrahydrofuran (5.6 mL). Water (5.6mL) and lithium hydroxide (0.050g, 2.07mmol) and methanol (1.2mL) were added. The reaction mixture was stirred at room temperature for 2.5 hours, after which the reaction mixture was concentrated under reduced pressure. Methylene chloride and aqueous hydrochloric acid (pH about 7) were added to the residue to obtain an emulsion. The reaction mixture was then concentrated to dryness under reduced pressure. The crude material was purified by supercritical fluid chromatography to 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶The corresponding epimers 1 and 2 of isothiazolidine-3-carboxylic acid (0.084 g (epimer 1) and 0.089g (epimer 2), 99% (combined yield), respectively).

Preferred preparation of the compounds of the invention

For reductive alkylation of 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]General procedure for hept-3-yl) -amides

Method A. sodium triacetoxyborohydride (purchased from Aldrich; 1.5 equivalents) was added to 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl at 0 deg.C]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 7, 1.0 equiv.) and aldehyde (1.2 equiv.) were added to a stirred solution in methanol (25 mL/mmol). The reaction mixture was stirred at rt for 16h and monitored by LCMS. The reaction mixture was then evaporated completely under reduced pressure, diluted with ethyl acetate (30mL/mmol) and washed with water (15 mL/mmol). The ethyl acetate layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude reaction mixture was purified by preparative HPLC to afford the desired compound.

Method B. conversion of 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl at 0 ℃]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (middle)Intermediate 6, 1.0 eq) and aldehyde (1.2 eq) to a stirred solution in methanol (25ml/mmol) was added sodium cyanoborohydride (purchased from Aldrich; 1.5 eq) and acetic acid (0.2 mL/mmol). The reaction mixture was stirred at rt for 16h and monitored by LCMS. The reaction mixture was then evaporated completely under reduced pressure, diluted with ethyl acetate (30mL/mmol) and washed with water (15 mL/mmol). The ethyl acetate layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude reaction mixture was purified by preparative HPLC to afford the desired compound.

Example 1

2- (3- { [ (5-chloro-2-hydroxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 5-chloro-2-hydroxy-benzaldehyde (purchased from Aldrich; 0.066g, 0.430mmol) to form 2- (3- { [ (5-chloro-2-hydroxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (0.0450g, 18%). Mass spectrum: m/Z: 701.5(M +1)

Example 2

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-iodo-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2-iodobenzaldehyde (purchased from Aldrich; 0.099g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-iodo-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.070g, 25%). Mass spectrum: m/Z: 777.4(M + 1).

Example 3

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-ethoxy-benzyl) -amino]Methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2-ethoxy-benzaldehyde (purchased from Aldrich; 0.0640g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-ethoxy-benzyl) -amino]Methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.050g, 21%). Mass spectrum: m/Z: 695.2(M + 1).

Example 4

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (5-isopropyl-2-methoxy-benzyl) -amino]-methyl } -benzyl) -1,1-1 lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 5-isopropyl-2-methoxy-benzaldehyde (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 0.0760g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (5-isopropyl-2-methoxy-benzyl) -amino]-methyl } -benzyl) -1,1-1 lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.075g, 29%). Mass spectrum: m/Z: 723.2(M + 1).

Example 5

2- (3- { [ (2-difluoromethoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2-difluoromethoxy-benzaldehyde (purchased from Aldrich; 0.0730g, 0.430mmol) to form 2- (3- { [ (2-difluoromethoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.061g, 24%). Mass spectrum: m/Z: 717.5(M + 1).

Example 6

2- [3- ({ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - [2- (4-fluoro-phenoxy) -benzyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2- (4-fluoro-phenoxy) -benzaldehyde (purchased from Aldrich; 0.093g, 0.430mmol) to form 2- [3- ({ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - [2- (4-fluoro-phenoxy) -benzyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.060g, 22%). Mass spectrum: m/Z: 761.5(M + 1).

Example 7

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-p-tolyloxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2- (4-methylphenoxy) benzaldehyde (purchased from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; 0.091g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-p-tolyloxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.080g, 30%). Mass spectrum: m/Z: 757.5(M + 1).

Example 8

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (4' -fluoro-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazoleAlkane-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2- (4-fluorophenyl) benzaldehyde (purchased from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; (0.086g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (4' -fluoro-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.045g, 17%). Mass spectrum: m/Z: 745.5(M + 1).

Example 9

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-hydroxy-4-methylbenzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2-hydroxy-4-methylbenzaldehyde (purchased from Aldrich Chemical Company, inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.058g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-hydroxy-4-methylbenzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.035g, 15%). Mass spectrum: m/Z: 681.6(M + 1).

Example 10

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-hydroxy-5-methyl-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2-hydroxy-5-methylbenzaldehyde (purchased from Aldrich Chemical Company, inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.058g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-hydroxy-5-methyl-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.050g, 21%). Mass spectrum: m/Z: 681.4(M + 1).

Example 11

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2' -methyl-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2' -methyl-biphenyl-2-carbaldehyde (purchased from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; 0.084g, 0.430mmol) was reacted to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2' -methyl-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.034g, 13%). Mass spectrum: m/Z: 741.6(M + 1).

Example 12

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-fluoro-6-phenoxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2-fluoro-6-phenoxybenzaldehyde (available from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; 0.093g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-fluoro-6-phenoxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.016g, 6%). Mass spectrum: m/Z: 761.6(M + 1).

Example 13

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2,3, 6-trisFluoro-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method A, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2,3, 6-trifluorobenzaldehyde (purchased from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; 0.068g, 0.430mmol) to form 2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2,3, 6-trifluoro-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.025g, 10%). Mass spectrum: m/Z: 705.5(M + 1).

Example 14

2- (3- { [ (6-bromo-2-hydroxy-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 6-bromo-2-hydroxy-3-methoxybenzaldehyde (available from Aldrich Chemical Company)Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.099g, 0.430mmol) to form 2- (3- { [ (6-bromo-2-hydroxy-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.055g, 20%). Mass spectrum: m/Z: 775.4(M + 1).

Example 15

2- (3- { [ (6-chloro-2-fluoro-3-methyl-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethylbutyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 6-chloro-2-fluoro-3-methylbenzaldehyde (available from Aldrich Chemical Company, inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.024g, 0.430mmol) to form 2- (3- { [ (6-chloro-2-fluoro-3-methyl-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethylbutyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.060g, 24%). Mass spectrum: m/Z: 717.4(M + 1).

Example 16

2- (3- { [ [2- (4-cyano-phenoxy) -benzyl]- ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl radical} -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 4- (2-formylphenoxy) benzonitrile (purchased from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; 0.096g, 0.430mmol) to form 2- (3- { [ [2- (4-cyano-phenoxy) -benzyl]- ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.024g, 9%). Mass spectrum: m/Z: 768.5(M + 1).

Example 17

2- (3- { [ benzo [1,3 ]]Dioxolen-4-ylmethyl- ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 2,3- (methylenedioxy) benzaldehyde (purchased from Aldrich Chemical Company, I)nc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.064g, 0.430mmol) to form 2- (3- { [ benzo [1,3 ] b]Dioxolen-4-ylmethyl- ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.038g, 15%). Mass spectrum: m/Z: 695.5(M + 1).

Example 18

2- (3- { [ (3-bromo-2, 6-difluoro-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt

Using method B, 2- {3- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (intermediate 6, 0.200g, 0.360mmol) and 3-bromo-2, 6-difluorobenzaldehyde (purchased from Alfa Aesar,26Parkridge Road, Ward Hill, MA 01835, USA; 0.095g, 0.430mmol) to form 2- (3- { [ (3-bromo-2, 6-difluoro-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate (0.038g, 15%). Mass spectrum: m/Z: 765.4(M + 1).

For coupling intermediate 8, 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶General method for producing (E) -isothiazolidine-3-carboxylic acid

In an ice bath(0 ℃) cooled 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶To a stirred solution of isothiazolin-3-carboxylic acid (intermediate 7, 1.0 equiv.) in dry N, N-dimethylformamide (8mL/mmol) were added O-benzotriazol-1-yl-N, N, N ', N' -tetramethyluronium hexafluorophosphate (available from Aldrich; 1.2 equiv.), 1-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 1.2 equiv.), amine (1.2 equiv.) and triethylamine (6.0 equiv.) in succession. The reaction mixture was allowed to warm to room temperature and stirred until the starting material was completely consumed (determined by LCMS). To the reaction mixture was added water and ethyl acetate. The organic phase was extracted with water and washed, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude reaction was purified by preparative HPLC to afford the desired compound as a TFA salt. The pure residue was taken up in ethyl acetate and washed with saturated sodium bicarbonate to remove TFA, then with water.

The above-described method was the method used in examples 19, 20, 24 to 28, and 30 to 34.

Example 19

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,5S) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid (intermediate 7, 0.089g, 0.148mmol) with (1R,2R,3R,5S) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-ylamine (available from Aldrich chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 30 μ L, 0.179mmol) to form 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,5S) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (0.075g, 68%). M/z =733(M + 1); HRMS: c₃₉H₅₉ClFN₄O₄S.Calc[M+H+]Calculated 733.3924, found 733.3920.

Example 20

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-1-ylamide

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of (E) -isothiazolidine-3-carboxylic acid (intermediate 7, 0.080g, 0.134mmol) with adamantan-1-amine (available from Aldrich Chemical Company, Inc.,1001 WestSaint Paul Avenue, Milwaukee, WI53233, USA; 0.025g, 0.179mmol) to form 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid adamantan-1-ylamide (0.066g, 68%). M/z =731(M + 1); HRMS: c₃₉H₅₇ClFN₄O₄S.Calc[M+H+]Calculated 731.3768, found 731.3766.

Example 21

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-first of allPhenyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 21mg, 0.11mmol) was added to 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl at 0 ℃ under argon]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amide (intermediate 5; 58mg, 0.1mmol) in methanol (3 mL). The mixture was stirred for a few minutes, then sodium triacetoxyborohydride (32mg, 0.15mmol) was added. The mixture was stirred at room temperature overnight. At this point, LCMS showed mainly starting material. The mixture was cooled in an ice bath and sodium cyanoborohydride (3mg, 0.05mmol) was added. The mixture was stirred for 2 h. LCMS showed about 50% conversion was complete. 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 10mg, 0.05mmol) was added followed by sodium cyanoborohydride (3mg, 0.05mmol) and acetic acid (50 μ L). The reaction mixture was stirred at room temperature for 4h, then concentrated under reduced pressure to remove methanol. Ethyl acetate (50mL) was added and the mixture was washed with water (2X 25mL) and brine (25 mL). The organic phase was dried over sodium sulfate, filtered and concentrated to give the crude product (78 mg). Purification by chromatography on a 4gm Isco column (eluting with 0-10% methanol/dichloromethane) followed by trituration with ether/pentane (1/1, 2mL) gave 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide as a white solid (18mg, 24%). HRMS: c₄₁H₅₅ClFN₄O₄S.Calc[M+H⁺]753.3611 calculated, found 753.3605.

Example 22

2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Step 1: (S) -2-amino-3- (3-chloro-phenyl) -N, N-dimethyl-propionamide trifluoroacetate salt

Boc-L-4-chlorophenylalanine (Chem-Impex International, Inc., Wood Dale, IL, USA; 5.0g, 16.7mmol), dimethylamine (2M in tetrahydrofuran; 12.5mL, 25mmol), 1-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 2.69g, 19.9mmol), O-benzotriazol-1-yl-N, N, N ', N' -tetramethyluronium hexafluorophosphate (available from Aldrich Chemical Company, Inc.,1001 Sast int Paul Avenue, Weukwaee, Wis 53233, USA; 7.5g, 19.8mmol) and diisopropylethylamine (available from Aldrich Chemical Company, 1001, Weuk, Miluk 53233, USA; 7.5g, 19.8mmol) and diisopropylethylamine (available from Aldrich Chemical Company, Inc., USA; Avenu 538, Avenu, Inc, USA; Avenu, Inc, W233, USA; Avenu, Inc, USA, N, Inc, W, Inc, N, W, Na, USA) was stirred at 0 ℃ for 30 min. Ethyl acetate was added and the mixture was washed with brine. The organic phase was dried over sodium sulfate, filtered and evaporated. The residue was dissolved in dichloromethane (80 mL). Trifluoroacetic acid (80mL) was added and the solution was stirred at room temperature for 2 h. Evaporation of the solvent gave (S) -2-amino-3- (3-chloro-phenyl) -N, N-dimethyl-propionamide trifluoroacetate (4.0g) as a white solid.

Step 2: (S) -3- (3-chloro-phenyl) -N1, N1-dimethyl-propane-1, 2-diamine hydrochloride

A solution of lithium aluminium hydride in tetrahydrofuran (3.5M; 20 mL; 70mmol) was carefully added to an ice-bath cooled solution of (S) -2-amino-3- (3-chloro-phenyl) -N, N-dimethyl-propionamide trifluoroacetate (4.00g, 11.8mmol) in tetrahydrofuran (20 mL). The reaction mixture was heated at reflux for 4h and then cooled. Ethyl acetate and diethyl ether were added followed by 3M sodium hydroxide solution (100 mL). The layers were separated and the aqueous layer was back-extracted with ether. The combined organic layers were washed with brine, dried over sodium sulfate, filtered and evaporated. The residue was taken up in a solution of 1M HCl in ether (20 mL). The resulting white precipitate was stirred for 1h and then filtered to give (S) -3- (3-chloro-phenyl) -N1, N1-dimethyl-propane-1, 2-diamine hydrochloride (1.92g, 57%).

And step 3: 2- {3- [ ((S) -3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

Triethylamine (338 μ L, 2.4mmol) was added to a solution of (S) -3- (3-chloro-phenyl) -N1, N1-dimethyl-propane-1, 2-diamine hydrochloride (264mg, 1.06mmol) in methanol (2mL) at room temperature under argon. The mixture was stirred for 10min, then 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda. was added under argon⁶A solution of ethyl-isothiazolidine-3-carboxylate (intermediate 8 step 3; 250mg, 0.96mmol) in methanol (4 mL). The mixture was stirred at room temperature for 15min, then cooled in an ice bath. Solid sodium cyanoborohydride (90.9mg, 1.45mmol) and glacial acetic acid (750 μ L) were added. The reaction mixture was allowed to warm to room temperature and stirred. The progress of the reaction was monitored by LCMS. When the reaction was substantially complete, the solvent was evaporated and ethyl acetate (50mL) was added. The solution was washed with water (2X 10mL)And the aqueous layer was extracted with ethyl acetate (50 mL). The combined organics were dried over sodium sulfate, filtered and concentrated to give crude 2- {3- [ ((S) -3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Ethyl isothiazolidine-3-carboxylate (430mg) as a yellow foam, which was used directly in the next step without further purification.

And 4, step 4: 2- {3- [ ((S) -3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶3-carboxylic acid (isothiazolidine)

To crude 2- {3- [ ((S) -3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶To a solution of ethyl-isothiazolidine-3-carboxylate (430mg, ca. 0.96mmol) in tetrahydrofuran (4mL) was added lithium hydroxide hydrate (71mg, 1.7mmol) followed by water (1 mL). The reaction mixture was stirred at room temperature for 2h, then the mixture was concentrated under reduced pressure to remove tetrahydrofuran. The residual solution was stirred and approximately 25 drops of 1M hydrochloric acid were added to bring the pH to 4.5. The sample was stirred for 10min and the aqueous solution was decanted from the yellow gum. The gum was washed with water (2 mL). The combined aqueous layers were evaporated to dryness and dried overnight to give 2- {3- [ ((S) -3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Isothiazolidine-3-carboxylic acid as a white residue (200 mg).

And 5:2- {3- { [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amides

Under argon gas, the(+) -Isopinocamphenylamine (Aldrich; 61mg, 0.38mmol), O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate (available from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 122mg, 0.38mmol), N-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 51mg, 0.38mmol) in anhydrous N, N-dimethylformamide (6mL) was added to a solution of 2- {3- [ ((S) -3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl-formamide]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid (200mg of about 90% pure, 0.38mmol) in a cooled (0 ℃ C.) solution. N, N-diisopropylethylamine (147. mu.L, 0.83mmol) was then added. The solution was then stirred at room temperature for 45 min. The reaction mixture was diluted with ethyl acetate (50mL) and washed with 1N NaOH (3X 15mL) and brine (20 mL). The organic phase was dried over sodium sulfate, filtered and concentrated to give the crude product (160mg) as a viscous oil. This material was combined with a mixture of 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶The crude products of the same experiment with isothiazolidine-3-carboxylic acid (270 mg; prepared in step 5 of the above-described process for the preparation of intermediate 6) were combined. The combined different batches of product were purified by chromatography (eluting with 0-10% methanol/dichloromethane) to give 2- {3- { [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amide (106mg, 18%) as a tan foam.

Step 6:2- {3- { [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amides

6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 18mg, 0.095 mmo) at 0 ℃ under argonl) was added to 2- {3- { [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amide (50mg, 0.086mmol) in methanol (3 mL). The mixture was stirred at 0 ℃ for a few minutes, then sodium cyanoborohydride (8mg, 0.13mmol) and glacial acetic acid (43 μ L) were added. The mixture was stirred at room temperature for 90 min. The solution was cooled again to 0 ℃ and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 5mg, 0.027mmol) was added. After 5min, sodium cyanoborohydride (5mg, 0.08mmol) from a new vial was added. The mixture was stirred at rt for 6 h. The solution was cooled again to 0 ℃ and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 5mg, 0.027mmol) was added. After 5min, sodium cyanoborohydride (5mg, 0.08mmol) was added. The mixture was stirred at room temperature overnight. The solution was cooled again to 0 ℃ and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 5mg, 0.027mmol) was added. After 5min, sodium cyanoborohydride (5mg, 0.08mmol) was added. The mixture was stirred at room temperature for 4 h. The mixture was concentrated to remove methanol. Ethyl acetate (50mL) was added and the mixture was washed with water (2X 25mL) and brine (25 mL). The aqueous layer was back-extracted with ethyl acetate (50 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated to give the crude product (82 mg). Purification by chromatography on an 8gm silica gel column (eluted with 0-10% methanol/dichloromethane) followed by trituration with ether/pentane (1/1, 2mL) gave 2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (3-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide as a white solid (40mg, 62%). HRMS: c₄₁H₅₄Cl₂FN₄O₄S.Calc[M+H⁺]Calculated 787.3222, found 787.3221.

Example 23

2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino }-methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Step 1: (S) -2-amino-3- (4-chloro-phenyl) -N, N-dimethyl-propionamide trifluoroacetate salt

A mixture of Boc-L-4-chlorophenylalanine (Bachem California Inc., Torrance, CA, USA; 5.0g, 16.7mmol), dimethylamine (2M in tetrahydrofuran; 12.5mL, 25mmol), 1-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 2.69g, 19.9mmol), O-benzotriazol-1-yl-N, N, N ', N' -tetramethyluronium hexafluorophosphate (available from Aldrich chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 7.5g, 19.8mmol) and diisopropylethylamine (8.8mL, 50.5mmol) in N, N-dimethylformamide was stirred at 0 ℃ for 30min and then at room temperature for 10 min. Ethyl acetate was added and the mixture was washed with water and brine. The organic phase was dried over sodium sulfate, filtered and evaporated to give a white solid. The solid was dissolved in dichloromethane (30 mL). Trifluoroacetic acid (30mL) was added and the solution was stirred at room temperature for 2 h. The solvent was evaporated to give (S) -2-amino-3- (4-chloro-phenyl) -N, N-dimethyl-propionamide trifluoroacetate as a white solid.

Step 2: (S) -3- (4-chloro-phenyl) -N1, N1-dimethyl-propane-1, 2-diamine hydrochloride

A solution of lithium aluminium hydride in tetrahydrofuran (Fluka; 3.5M; 23 mL; 80.5mmol) was carefully added to an ice-bath cooled solution of (S) -2-amino-3- (4-chloro-phenyl) -N, N-dimethyl-propionamide trifluoroacetate (3.00g, 13.2mmol) in anhydrous tetrahydrofuran (36 mL). The reaction mixture was heated at reflux for 4h and then cooled with an ice bath. Ethyl acetate (25mL) was carefully added to consume excess lithium aluminum hydride, followed by 3M sodium hydroxide solution (25 mL). Ethyl acetate (100mL) and saturated brine (50mL) were added and the mixture was stirred vigorously. The layers were separated and the organic layer was washed with brine (50 mL). The brine extract was back-extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, evaporated, and concentrated to give the crude free base (1.9 g). It was taken up in anhydrous ether (25mL) and the mixture was stirred. A solution of HCl in ether (1M; 25mL) was added dropwise. The resulting white precipitate was stirred for 5min, filtered under a stream of nitrogen, washed with diethyl ether (3 × 5mL), and then dried under high vacuum to give (S) -3- (4-chloro-phenyl) -N1, N1-dimethyl-propane-1, 2-diamine hydrochloride (2.1g, 64%) as a light yellow hygroscopic solid.

And step 3: 2- {3- [ ((S) -4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

Triethylamine (214. mu.L, 1.5mmol) was added to a solution of (S) -3- (4-chloro-phenyl) -N1, N1-dimethyl-propane-1, 2-diamine hydrochloride (intermediate X; 182mg, 0.73mmol) in methanol (2 mL). The mixture was stirred for 10min, then 2- (3-formyl-benzyl) -1, 1-dioxo-1. lamda. was added under argon⁶A solution of ethyl-isothiazolidine-3-carboxylate (intermediate 8, step 3; 190mg, 0.61mmol) in methanol (4 mL). The mixture was stirred at room temperature for 15min, then cooled in an ice bath. Solid sodium cyanoborohydride (57.5mg, 0.9mmol) and glacial acetic acid (540. mu.L) were added. The reaction mixture was allowed to warm to room temperature and stirred. The progress of the reaction was monitored by LCMS. When the reaction is substantially complete, the solvent is evaporated and acetic acid is addedEthyl ester (50 mL). The solution was washed with water (2X 10mL) and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organics were dried over sodium sulfate, filtered and concentrated to give crude 2- {3- [ ((S) -4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Ethyl isothiazolidine-3-carboxylate (390mg), a sticky yellow foam, which was used directly in the next step without further purification.

And 4, step 4: 2- {3- [ ((S) -4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶3-carboxylic acid (isothiazolidine)

To crude 2- {3- [ ((S) -4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶To a solution of ethyl-isothiazolidine-3-carboxylate (310mg, ca. 0.61mmol) in tetrahydrofuran (3.6mL) was added lithium hydroxide hydrate (51mg, 1.2mmol) followed by water (910. mu.L). The reaction mixture was stirred at room temperature for 2h, then the mixture was concentrated under reduced pressure to remove tetrahydrofuran. The residual solution was stirred and approximately 25 drops of 1M hydrochloric acid were added to bring the pH to 4.5. The sample was stirred for 10min and the aqueous solution was decanted from the yellow gum. The gum was washed with water (2 mL). The combined aqueous layers were evaporated to dryness and dried overnight to give 2- {3- [ ((S) -4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶Isothiazolidine-3-carboxylic acid as a pale yellow residue (292 mg).

And 5:2- {3- { [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amides

A solution of (+) -isopinocampheylamine (Aldrich; 88mg, 0.55mmol), O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate (available from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 177mg, 0.55mmol), N-hydroxybenzotriazole (available from 3B Scientific Corporation, Libertyville, IL60048, USA; 74mg, 0.55mmol) in anhydrous N, N-dimethylformamide (8mL) was added to 2- {3- [ ((S) -4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl-formamide (8mL) under argon]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid (292mg about 90% pure, 0.55mmol) in cooled (0 ℃ C.) solution. N, N-diisopropylethylamine (214. mu.L, 1.2mmol) was then added. The solution was then stirred at room temperature for 45 min. The reaction mixture was diluted with ethyl acetate (50mL) and washed with 1N NaOH (3X 15mL) and brine (20 mL). The organic phase was dried over sodium sulfate, filtered and concentrated to give the crude product (370mg) as a viscous oil. This material was purified by chromatography (eluting with 0-10% methanol/dichloromethane) to give 2- {3- { [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amide (110mg, 33%) as a pale tan hygroscopic foam.

Step 6:2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 37mg, 0.2mmol) was added to 2- {3- { [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethylamino) -methyl at 0 deg.C under argon]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethylYl-bicyclo [3.1.1]Hept-3-yl) amide (110mg, 0.18mmol) in methanol (3 mL). The mixture was stirred at 0 ℃ for a few minutes, then sodium cyanoborohydride (17mg, 0.27mmol) and glacial acetic acid (95. mu.L) were added. The mixture was stirred at room temperature overnight. The solution was cooled to 0 ℃ again and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 4mg, 0.02mmol) was added followed by sodium cyanoborohydride (3mg, 0.05 mmol). The mixture was stirred for 2 h. The solution was cooled again to 0 ℃ and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 5mg, 0.027mmol) was added followed by sodium cyanoborohydride (5mg, 0.08 mmol). The mixture was stirred at room temperature for 4 h. The solution was cooled again to 0 ℃ and 6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 5mg, 0.027mmol) was added followed by sodium cyanoborohydride (5mg, 0.08 mmol). The mixture was stirred at rt for 6 h. The mixture was concentrated to remove methanol. Ethyl acetate (50mL) was added and the mixture was washed with water (2X 25mL) and brine (25 mL). The aqueous layer was back-extracted with ethyl acetate (50 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated to give the crude product. This material was purified by chromatography on an 8gm silica gel column (eluting with 0-10% methanol/dichloromethane) followed by trituration with ether/pentane (1/1, 2mL) to give 2- [3- ({ (6-chloro-2-fluoro-3-methoxy-benzyl) - [ (S) -2- (4-chloro-phenyl) -1-dimethylaminomethyl-ethyl]-amino } -methyl) -benzyl]-1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide as a white solid (32mg, 25%). HRMS: c₄₁H₅₄Cl₂FN₄O₄S.Calc[M+H⁺]Calculated 787.3222, found 787.3221.

Example 24

[2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- (4-phenyl-piperidin-1-yl) -methanones

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of (E) -isothiazolidine-3-carboxylic acid (intermediate 7, 0.065g, 0.108mmol) with 4-phenylpiperidine (purchased from Aldrich Chemical Company, Inc.,1001 WestSaint Paul Avenue, Milwaukee, WI53233, USA; 0.023g, 0.140mmol) to form [2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- (4-phenyl-piperidin-1-yl) -methanone (0.018g, 23%). M/z =741(M + 1); HRMS: c₄₀H₅₅ClFN₄O₄S.Calc[M+H⁺]Calculated 741.3611, found 741.3611.

Example 25

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (epimer 1)

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid (intermediate 7 (epimer 1), 0.070g, 0.117mmol) and (1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [ 3.1.1)]Hept-3-ylamine (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 25. mu.L, 0.148mmol) was reacted to form 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-bisOxo-1 lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (epimer 1) (0.062g, 72%). M/z =733(M + 1); HRMS: c₃₉H₅₉ClFN₄O₄S.Calc[M+H⁺]Calculated 733.3924, found 733.3925.

Example 26

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (epimer 2)

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid (intermediate 7 (epimer 2), 0.075g, 0.125mmol) and (1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [ 3.1.1)]Hept-3-ylamine (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 26. mu.L, 0.154mmol) was reacted to form (R) -2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (epimer 2) (0.052g, 57%). M/z =733(M + 1); HRMS: c₃₉H₅₉ClFN₄O₄S.Calc[M+H⁺]Calculated 733.3924, found 733.3925.

Example 27

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶3-carboxylic acid 4-chloro-benzylamide

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of (E) -isothiazolidine-3-carboxylic acid (intermediate 7, 0.085g, 0.142mmol) with 4-chlorobenzylamine (purchased from Aldrich Chemical Company, Inc.,1001 WestSaint Paul Avenue, Milwaukee, WI53233, USA; 43. mu.L, 0.355mmol) to form 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶4-chloro-benzylamide (0.038g, 37%) of isothiazolidine-3-carboxylic acid. M/z =721(M + 1); HRMS: c₃₆H₄₈Cl₂FN₄O₄S.Calc[M+H⁺]Calculated 721.2752, found 721.2755.

Example 28

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((S) -1,7, 7-trimethyl-bicyclo [ 2.2.1)]Hept-2-yl) -amides

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid (intermediate 7, 0.090g, 0.150mmol) and (R) - (+) -bornylamine (available from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.034g, 0225mmol) to form 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((S) -1,7, 7-trimethyl-bicyclo [ 2.2.1)]Hept-2-yl) -amide (0.053g, 49%). M/z =733(M + 1); HRMS: c₃₉H₅₉ClFN₄O₄S.Calc[M+H⁺]Calculated 733.3924, found 733.3927.

Example 29

2- (3- { [ (2, 3-difluoro-6-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Step 1:2, 3-difluoro-6-methoxy-benzaldehyde

A solution of 1, 2-difluoro-4-methoxy-benzene (Aldrich; 10.0g, 69.4mmol) in anhydrous tetrahydrofuran (500mL) was cooled to-78 ℃ under a nitrogen atmosphere. Lithium diisopropylamide solution (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, Wis.53233, USA; 1.8M solution in tetrahydrofuran/heptane/ethylbenzene, 40mL, 72.9mmol) was added dropwise with a syringe. The reaction mixture was warmed to-55 ℃ and held at this temperature for 1 h. The mixture was then cooled again to-78 ℃ and dry N, N-dimethylformamide (10.7mL, 139mmol) was added via syringe. The cooling bath was removed and the reaction mixture was allowed to warm to-10 ℃ and quenched by the addition of flake ice (about 200mL) and saturated ammonium chloride solution (200 mL). Ethyl acetate (200mL) was added, the layers were separated and the aqueous layer was extracted with ethyl acetate (200 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, evaporated, and purified by silica gel chromatography (eluting with 0-30% ethyl acetate/hexanes) to give 2, 3-difluoro-6-methoxy-benzaldehyde (7.0g, 59% yield) as an oil that solidified upon standing.

Step 2:2- (3- { [ (2, 3-difluoro-6-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

2, 3-difluoro-6-methoxy-benzaldehyde (from step 1; 29mg, 0.17mmol) was added to 2- {3- [ ((S) -1-dimethylaminomethyl-2-phenylethylamino) -methyl at room temperature under argon]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) amide (intermediate 5; 90mg, 0.16mmol) in methanol (3 mL). The mixture was stirred for a few minutes, then sodium cyanoborohydride (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 5mg, 0.08mmol) and glacial acetic acid (78 μ L) were added. The mixture was stirred at room temperature for 4 h. 2, 3-difluoro-6-methoxy-benzaldehyde (from step 1; 5mg, 0.03mmol) was added followed by sodium cyanoborohydride (3mg, 0.05 mmol). The mixture was stirred at room temperature for 4 h. The mixture was stirred at room temperature overnight, then concentrated under reduced pressure to remove methanol. Ethyl acetate (50mL) was added and the mixture was washed with water (2X 25mL) and brine (25 mL). The aqueous layer was back-extracted with ethyl acetate (50 mL). The combined organic phases were dried over sodium sulfate, filtered and concentrated to give the crude product. Purification by chromatography on a 12gm silica gel column (eluting with 0-10% methanol/dichloromethane) followed by trituration with ether/pentane (1/1, 2mL) gave a partially purified material. Ethyl acetate (3mL) and saturated sodium bicarbonate solution (2mL) were added and the mixture was stirred for 30 min.The mixture was extracted with ethyl acetate (2 × 25mL) and the organic layer was washed with brine, then dried, filtered and evaporated to give 2- (3- { [ (2, 3-difluoro-6-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide (48mg, 42%) as a white solid. HRMS: c₄₁H₅₅F₂N₄O₄S.Calc[M+H⁺]Calculated 737.3907, found 737.3906.

Example 30

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-2-ylamide

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of (i) -isothiazolidine-3-carboxylic acid (intermediate 7, 0.093g, 0.155mmol) with 2-adamantanamine hydrochloride (available from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.036,0.192mmol) to form 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid adamantan-2-ylamide (0.075g, 66%). M/z =731(M + 1); HRMS: c₃₉H₅₇ClFN₄O₄S.Calc[M+H⁺]Calculated 731.3768, found 731.3773.

Example 31

2- (3- { [ (6-chloro-2-fluoro-3-methyl)Oxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,4S) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amide and 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,4R) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amides

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid (intermediate 7, 0.085g, 0.144mmol) and (+/-) -2-endo-amino-3-exo-isopropylbicyclo [2.2.1]Heptane (Sigma-Aldrich corporation, St. Louis, MO, USA; 0.041g, 0.216mmol) reacted to form: 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,4S) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amide and 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,4R) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amide (0.048g, 45%) as a mixture of diastereomers without separation. M/z =733(M + 1); HRMS: c₃₉H₅₉ClFN₄O₄S.Calc[M+H⁺]Calculated 733.3924, found 733.3924.

Example 32

[2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- [4- (4-chloro-phenyl) -4-hydroxy-piperazinePyridin-1-yl]-methanones

Reacting 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of (E) -isothiazolidine-3-carboxylic acid (intermediate 7, 0.081g, 0.135mmol) with 4- (4-chlorophenyl) -4-hydroxypiperidine (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.035,0.166mmol) to form [2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- [4- (4-chloro-phenyl) -4-hydroxy-piperidin-1-yl group]-methanone (0.066g, 62%). M/z =791(M + 1); HRMS: c₄₀H₅₄Cl₂FN₄O₅S.Calc[M+H⁺]Calculated 791.3171, found 791.3169.

Example 33

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-1-ylamide (epimer 1)

Reacting (S) -2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of (S) -2- (3- { [ (6-chloro-2-fluoro-6-fluoro-2-fluoro-) -isothiazolidine-3-carboxylic acid (intermediate 7 (epimer 1), 0.071g, 0.119mmol) with adamantan-1-amine (available from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 0.022g, 0.144mmol) to form (S) -2- (3- { [ (6-chloro-2-fluoro)-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid adamantan-1-ylamide (epimer 1) (0.054g, 61%). M/z =731(M + 1); HRMS: c₃₉H₅₇ClFN₄O₄S.Calc[M+H⁺]Calculated 731.3768, found 731.3765.

Example 34

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid adamantan-1-ylamide (epimer 2)

Reacting (R) -2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Reaction of isothiazolidine-3-carboxylic acid (intermediate 7 (epimer 2), 0.044g, 0.073mmol) with adamantan-1-amine (0.013g, 0.089mmol) to form (R) -2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶Isothiazolidine-3-carboxylic acid adamantan-1-ylamide (epimer 2) (0.033g, 61%). M/z =731(M + 1); HRMS: c₃₉H₅₇ClFN₄O₄S.Calc[M+H⁺]Calculated 731.3768, found 731.3767.

Example 35

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Step 1: [3- (tert-butyl-dimethyl-siloxymethyl) -5-nitro-phenyl ] -methanol

A solution of tert-butyldimethylchlorosilane (from Aldrich chemical company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 1.3g, 8.6mmol) in dichloromethane (40mL) was added dropwise over a period of 2 hours at 0 ℃ to a magnetically stirred solution of (3-hydroxymethyl-5-nitro-phenyl) -methanol (from Aldrich; 1.50g, 8.19mmol) and imidazole (0.610g, 9.00mmol) in dichloromethane (80 mL). The resulting mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was washed with water (2 × 200mL) and brine, dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 10-75% ethyl acetate/hexanes) to give [3- (tert-butyl-dimethyl-siloxymethyl) -5-nitro-phenyl ] -methanol (1.20g, 49% yield) as a viscous oil that solidified upon standing.

Step 2: [ 3-amino-5- (tert-butyl-dimethyl-siloxymethyl) -phenyl ] -methanol

10% Pd/C (53mg) was added to [3- (tert-butyl-dimethyl-siloxymethyl) -5-nitro-phenyl]Methanol (535mg, 1.80mmol) in a magnetically stirred solution in ethanol (20 mL). The heterogeneous mixture was purged twice with vacuum and dry nitrogen and then stirred at room temperature under hydrogen at atmospheric pressure. After 3 hours, the reaction mixture is filtered through kieselguhrA pad. The pad was washed with ethanol (2X 20mL) and the filtrate was concentrated to give crude [ 3-amino-5- (tert-butyl-dimethyl-silanyloxymethyl) -phenyl]Methanol (515mg) as an off-white solid, which was used directly in the next step without purification.

And step 3: [3- (tert-butyl-dimethyl-siloxymethyl) -5-dimethylamino-phenyl ] -methanol

Sodium cyanoborohydride (470mg, 7.48mmol) was added to a magnetically stirred solution of [ 3-amino-5- (tert-butyl-dimethyl-silanyloxymethyl) -phenyl ] -methanol (1.0g, 3.74mmol) in methanol (30 mL). The reaction vessel was closed with a septum and pierced with a needle. In a separate flask, paraformaldehyde (449mg, 15.0mmol) was heated with a hot air gun and the resulting gas was introduced into the reaction vessel with a conduit with a stream of nitrogen. After all the paraformaldehyde was added, a solution of zinc chloride (510mg, 3.74mmol) in methanol (8mL) was added dropwise. The generation of gas was found. After the addition was complete, TLC showed that the starting material had been consumed. The reaction mixture was diluted with ethyl acetate (100mL) and washed with 1% aqueous ammonium hydroxide solution, then brine. The solution was dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 10-60% ethyl acetate/hexanes) to give [3- (tert-butyl-dimethyl-siloxymethyl) -5-dimethylamino-phenyl ] -methanol (906mg, 82% yield) as a viscous oil.

And 4, step 4: 3- (tert-butyl-dimethyl-siloxymethyl) -5-dimethylamino-benzaldehyde

A suspension of Daiss-Martin iodophor (available from Aldrich Chemical Company, Inc.,1001 WestSaint Paul Avenue, Milwaukee, WI53233, USA; 538mg, 1.27mmol) in dichloromethane (10mL) was pipetted into a magnetically stirred solution of [3- (tert-butyl-dimethyl-silanyloxymethyl) -5-dimethylamino-phenyl ] -methanol (288mg, 0.986mmol) in dichloromethane (20 mL). The mixture was stirred for 15 min. Powdered sodium bicarbonate (ca 500mg) was added and volatiles were removed in vacuo. The residue was purified by silica gel chromatography (eluting with 10-40% ethyl acetate/hexanes) to give 3- (tert-butyl-dimethyl-silanyloxymethyl) -5-dimethylamino-benzaldehyde (187mg, 65%) as a yellow oil.

And 5: 3-bromomethyl-5-dimethylamino-benzaldehyde

Glacial acetic acid (6mL) and water (2mL) were added to a magnetically stirred solution of 3- (tert-butyl-dimethyl-silanyloxymethyl) -5-dimethylamino-benzaldehyde (344mg, 1.2mmol) in tetrahydrofuran (2 mL). The mixture was stirred for 2h, then an additional 10mL of glacial acetic acid was added. The reaction mixture was stirred overnight. TLC showed the presence of trace amounts of starting material and a slower moving spot (moving spot). Volatiles were removed in vacuo and glacial acetic acid (24mL), water (8mL) and tetrahydrofuran (8mL) were added. The resulting mixture was warmed to 40 ℃. After 2.5 hours, TLC showed complete consumption of the starting material. The volatiles were removed in vacuo and the residue was dissolved in ethyl acetate/water (100mL 1:1 v/v). The aqueous layer was made alkaline with saturated aqueous sodium carbonate solution and the layers were separated. The aqueous layer was extracted with ethyl acetate (25 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to give the intermediate alcohol (178mg, 81%) as a pale white oil. This material was dissolved in tetrahydrofuran (10mL) and the resulting solution was added to a solution that had been prepared using the following method: bromine (102 μ L, 1.99mmol) was added dropwise to a magnetically stirred solution of triphenylphosphine (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 522mg, 1.99mmol) in 20mL of acetonitrile at 0 ℃ under a nitrogen atmosphere using a syringe and the solution was stirred for 10min at 0 ℃. The reaction mixture was stirred at about 0 ℃ for 20min, then water (50mL), a small amount of crystals of sodium bisulfite, and ethyl acetate (50mL) were added. The aqueous layer was separated and washed once with ethyl acetate (50 mL). The combined organic layers were dried over sodium sulfate, filtered, evaporated and purified by filtration through a plug of silica gel (plug) then eluted with 20% ethyl acetate/hexanes to give 3-bromomethyl-5-dimethylamino-benzaldehyde (100mg, 35% yield, over two steps).

Step 6:2- (3-dimethylamino-5-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶3-Thiazolidine-carboxylic acid methyl ester and 2- (3-dimethylamino-5-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

Reacting (S) -1, 1-dioxo-1. lamda⁶3-Thiazolidine-carboxylic acid methyl ester and (S) -1, 1-dioxo-1. lamda⁶A4: 3 mixture of-isothiazolidine-3-carboxylic acid ethyl ester benzaldehyde (see intermediate 8, step 2; 148mg, 0.84mmol) and cesium carbonate (134mg, 0.41mmol) was added to a magnetically stirred solution of 3-bromomethyl-5-dimethylamino-benzaldehyde (100mg, 0.413mmol) in anhydrous N, N-dimethylformamide (10 mL). The resulting suspension was stirred overnight. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate (25mL) and washed with water (50 mL). The aqueous layer was separated and extracted once with ethyl acetate (25 mL). The combined organic layers were dried over sodium sulfate, filtered, evaporated and purified by silica gel chromatography (eluting with 10-75% ethyl acetate/hexanes) to give (S) -2- (3-dimethylamino-5-formyl-benzyl) -1, 1-dioxo-1. lambda⁶Methyl (3-isothiazolidine) -carboxylate and (S) -2- (3-dimethylamino-5-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶A 1:1 mixture of ethyl (144mg) of isothiazolidine-3-carboxylate.

And 7:2- { 3-dimethylamino-5- [ ((a))(S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid methyl ester and 2- { 3-dimethylamino-5- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

(S) -4,4, N1, N1-tetramethyl-pentane-1, 2-diamine hydrochloride (intermediate 2, 0.100g, 0.43mmol), magnesium sulfate (100mg) and triethylamine (118. mu.L, 0.847mmol) were added to (S) -2- (3-dimethylamino-5-formyl-benzyl) -1, 1-dioxo-1. lambda⁶Methyl (3-isothiazolidine) -carboxylate and (S) -2- (3-dimethylamino-5-formyl-benzyl) -1, 1-dioxo-1. lamda.⁶A solution of a 1:1 mixture of ethyl-isothiazolidine-3-carboxylate (144mg) in anhydrous dichloromethane (15 mL). The resulting cloudy mixture was stirred at room temperature for 5 h. The volatiles were removed in vacuo and the residue was suspended in dry methanol (10 mL). Sodium cyanoborohydride (purchased from Aldrich Chemical Company, Inc.,1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 53mg, 0.85mmol) and acetic acid (85. mu.L) were added and the mixture was stirred overnight. The volatiles were removed in vacuo and the residue was dissolved in ethyl acetate (50 mL). The solution was washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and evaporated under reduced pressure to give 2- { 3-dimethylamino-5- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid methyl ester and 2- { 3-dimethylamino-5- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶A 1:1 mixture of ethyl isothiazolidine-3-carboxylate (155mg) which was used directly in the next step without further purification.

And 8:2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid methyl ester and2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ethyl ester

6-chloro-2-fluoro-3-methoxy-benzaldehyde (intermediate 4; 91mg, 0.482mmol) and magnesium sulfate (100mg) were added to 2- { 3-dimethylamino-5- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid methyl ester and 2- { 3-dimethylamino-5- [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butylamino) -methyl]-benzyl } -1, 1-dioxo-1. lamda.⁶A 1:1 mixture of ethyl-isothiazolidine-3-carboxylate (155mg) in a magnetically stirred solution in anhydrous dichloromethane (15 mL). The resulting cloudy mixture was stirred at room temperature for 5 h. The volatiles were removed in vacuo and the residue was suspended in dry methanol (16mL) and treated with sodium cyanoborohydride (40mg, 0.64mmol) and acetic acid (128. mu.L). The mixture was stirred overnight. The volatiles were removed in vacuo and the residue was dissolved in ethyl acetate (50 mL). The solution was washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and evaporated under reduced pressure to give 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid methyl ester and 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶A 1:1 mixture of ethyl isothiazolidine-3-carboxylate (265mg, isolated), which was used directly in the next step without further purification.

And step 9: 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amides

Lithium hydroxide (15mg, 0.61mmol) was added to 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid methyl ester and 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶A 1:1 mixture of ethyl-isothiazolidine-3-carboxylate (265mg) in a solution of methanol (15mL) and water (5 mL). The reaction mixture was stirred at room temperature for 3h, then concentrated to dryness under reduced pressure. Anhydrous N, N-dimethylformamide (available from Aldrich Chemical Company, Inc.,1001 WestSaint Paul Avenue, Milwaukee, WI53233, USA; 15mL) was added to the residue, followed by O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (available from Aldrich Chemical Company, Inc., West Saint Paul Avenue, Milwaukee, WI53233, USA; 184mg, 0.49mmol), 1-hydroxybenzotriazole (available from 3B Scientific Corporation, Liberville, IL60048, USA; 74mg, 0.49mmol), (1S,2S,3S,5R) - (+) -isopinopinene amine (available from Aldrich Company, Inc.,1001, Miluk, Wik, Avenune 53233, USA; 20 μm, USA; 5 U.5R) - (+) -isopinopinene amine (available from Aldrich Chemical Company, Inc., USA; 5 μm; 9 μm; isopropyl amine, Inc., USA; 5 μ g; 5 μ M; 5 U., 1001West Saint Paul Avenue, Milwaukee, WI53233, USA; 93 μ L, 0.45 mmol). The resulting mixture was stirred overnight. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate (50mL) and washed with water (50 mL). The aqueous layer was back-extracted with ethyl acetate (50mL) and the combined organic layers were dried over sodium sulfate, filtered and concentrated. The crude product was purified by HPLC to give 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) Amide (300mg, about 91%, subjected to 5 steps). HRMS: c₄₁H₆₄ClFN₅O₄S.Calc[M+H⁺]Calculated 776.4346, found 776.4348.

Example 36

Testing of Compounds of the invention

Promega's CellTiter-Cytotoxicity was evaluated by luminescence cell viability assay (Promega Corporation, Madison, WI, USA). The assay was performed in HeLa cells at a maximum test concentration of 10 μ M test compound. The assay measures the amount of Adenosine Triphosphate (ATP), which represents a means of quantitatively measuring the presence and extent of metabolic activity in cell cultures.

Viable Hela cells were plated at a density of 3000 cells/well in a 96-well black, transparent bottom plate. Cells were attached to the plate by overnight incubation at 37 ℃. Test compounds were diluted in DMSO and then further diluted in 10% FBS medium. Media was removed from the cell plate and 50 μ Ι _ of solution was added manually to each well except for the control wells (which contained the same volume of 10% FBS media containing 1% DMSO). Cells were incubated undisturbed for 48 hours at 37 ℃ in the presence of compound. After 48 hours exposure to the compound, 50. mu.L of pCellTiter was addedAnd (3) a reagent. The cell plate was shaken on a shaker for 2 minutes to aid in cell lysis. The plate was covered and read for luminescence. The percent inhibition of luminescence compared to control cells was measured for each compound and is shown in the table below.

Percent inhibition was calculated as follows:

PCT_INHIB=100*[(U-T)/U]

wherein U represents luminescence of a well containing untreated cells and

t represents luminescence of wells containing compound-treated cells

Example 37

MTT tetrazolium dye proliferation assay

Proliferation of adherent cell lines was assessed by detection of tetrazolium dye according to the methods of Denizot and Lang (F. Denizot and R. Lang J. Immunol. methods1986,89, 271-277). The cell line used was H196, namely from ATCC (ATCC number CRL-5823)^TM) The obtained small cell lung cancer cell line. The cells were grown in RPMI 1640 medium supplemented with 2.5% fetal bovine serum.

Cells were seeded at 20,000 cells/well in clear 96-well tissue culture treatment plates. The plates were mixed in a 5% CO₂Incubated overnight at 37 ℃. The following day, test compounds were serially diluted in appropriate growth media to three times the final concentration. 50 μ L of 3 Xdiluted drug was added to each well to give a final volume of 150 μ L/well. Six concentrations (10. mu.M to 30nM) were tested for each compound. Control wells (without inhibitor) had a final DMSO concentration of 0.25%. 50 μ L of 0.75% DMSO in growth medium was added to each well to a final volume of 150 μ L/well. The plate was returned to the incubator and after 5 days the plate was analysed by the following method: 3- (4, 5-Dimethylthiazol-2-yl) -2, 5-biphenyl-2H-tetrazolium bromide (thiazole blue; MTT; Sigma) was added to each well to give a final concentration of 1.25 mg/mL. The plate was returned to the incubator for 3 hours. Removing the MTT-containing medium and subjecting the resulting formazan toThe (formazan) metabolite was dissolved in 100% ethanol and shaken at room temperature for 10 min. The absorbance was read at a wavelength of 570nm (referenced at 660 nm) on a plate reader. The concentrations of compounds that resulted in 50% and 90% inhibition of cell proliferation were determined from a plot of the logarithm of the concentration versus the percentage inhibition using the curve fitting function of Microsoft Excel. These concentrations represent IC, respectively₅₀And IC₉₀The value is obtained.

Example 38

MTS tetrazolium assay

Proliferation of non-adherent cell lines was assessed using the MTS tetrazolium dye assay according to the method of Malich and co-workers (G.Malich et al. society 1997,124, 179-192). The cell line used was H146(ATCC No. HTB-173)^TM). The cells were grown in RPMI 1640 medium supplemented with 2.5% fetal bovine serum.

Cells were seeded at 20,000 cells/well in clear 96-well tissue culture treatment plates. Subjecting said panel to a reaction in the presence of 5% CO₂Incubated overnight at 37 ℃. The next day, test compounds were continued in appropriate growth mediumDiluted to three times the final concentration. 50 μ L of 3 × diluted drug was added to each well to a final volume of 150 μ L/well. Control wells (without inhibitor) had a final DMSO concentration of 0.25%. 50 μ L of 0.75% DMSO in growth medium was added to each well to a final volume of 150 μ L/well. The plate was returned to the incubator. After 5 days, CellTiter was addedAQueous One solution (Promega Corporation, Madison, Wis., USA) (25. mu.L/well) was added to each well and the plate was returned to the incubator for 2-3 h. The absorbance at 490nm was measured for each well using a plate reader. The concentrations of compounds that resulted in 50% and 90% inhibition of cell proliferation were determined from a plot of the logarithm of the concentration versus the percentage inhibition using the curve fitting function of Microsoft Excel. These concentrations represent IC, respectively₅₀And IC₉₀The value is obtained.

Claims

1. A compound of formula 1 or a pharmaceutically acceptable salt thereof,

wherein the compound exhibits cytotoxic activity and:

R₂is hydrogen or dimethylamino;

R₃is hydrogen;

R₄is optionally substituted with C_1-6Alkyl radicalA bicyclic or tricyclic cycloalkyl group having 7 to 10 ring atoms, or

Benzyl optionally substituted with halogen;

R₅is C_1-20Alkyl or benzyl optionally substituted with halogen; and

R₆is benzodioxolyl or phenyl optionally substituted with 1 to 3 substituents independently selected from the group consisting of: halogen, hydroxy, C optionally substituted by halogen_1-20Alkoxy radical, C_1-20Alkyl, optionally substituted with halogen, cyano or C_1-20Phenoxy radical of alkyl and optionally substituted with C_1-20Alkyl or halogen phenyl.

2. The compound of claim 1, wherein R₂Is hydrogen.

3. The compound of claim 1, wherein R₂Is dimethylamino.

4. The compound of claim 1, wherein R₃Is hydrogen.

5. The compound of claim 1, wherein R₄Is optionally substituted with C_1-6A bicyclic or tricyclic cycloalkyl group having 7 to 10 ring atoms of the alkyl group.

6. The compound of claim 1, wherein R₄Is benzyl optionally substituted with halogen.

7. The compound of claim 1, wherein R₃And R₄Together with the nitrogen atom to which they are attached form a piperidinyl group substituted with one or more substituents independently selected from the group consisting of: hydroxy and optionally substitutedPhenyl with halogen.

8. The compound of claim 1, wherein R₅Is benzyl optionally substituted with halogen.

9. The compound of claim 1, wherein R₆Is phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, C optionally substituted by halogen_1-20Alkoxy radical, C_1-20Alkyl, optionally substituted with halogen, cyano or C_1-20Phenoxy radical of alkyl and optionally substituted with C_1-20Alkyl or halogen phenyl.

10. The compound of claim 1, wherein R₄Is bicycloheptyl substituted three times with methyl.

11. The compound of claim 1, wherein R₄Is adamantyl.

12. The compound of claim 1, wherein R₄Is bicyclo [3.1.1]Hept-3-yl.

13. The compound of claim 1, wherein R₅Is2, 2-dimethylpropyl.

14. The compound of claim 1, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

R₆is phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, C optionally substituted by halogen_1-20Alkoxy radical, C_1-20Alkyl, optionally substituted with halogen, cyano or C_1-20Phenoxy radical of alkyl and optionally substituted with C_1-20Alkyl or halogen phenyl.

15. The compound of claim 14 selected from the group consisting of:

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (2-p-tolyloxy-benzyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trisA fluoroacetate salt;

2- (3- { [ ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) - (4' -fluoro-biphenyl-2-ylmethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (6-bromo-2-hydroxy-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

2- (3- { [ (6-chloro-2-fluoro-3-methyl-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethylbutyl) -amino]-methyl } -benzyl) -1, 1-dioxoGeneration-1 lambda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide trifluoroacetate salt;

16. The compound of claim 1, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is adamantyl;

R₅is2, 2-dimethylpropyl; and

17. The compound of claim 16, selected from the group consisting of:

18. The compound of claim 1, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

R₆is benzodioxolyl.

19. The compound of claim 1, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is benzyl optionally substituted with halogen; and

20. The compound of claim 19 selected from the group consisting of:

2- (3- { [ (2, 3-difluoro-6-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-2-phenyl-ethyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide.

21. The compound of claim 1, wherein:

R₂is hydrogen;

R₅is2, 2-dimethylpropyl; and

22. The compound of claim 21, wherein the compound is:

[2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidin-3-yl]- (4-phenyl-piperidin-1-yl) -methanone; or

23. The compound of claim 1, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is benzyl optionally substituted with halogen;

R₅is2, 2-dimethylpropyl; and

24. The compound of claim 23, wherein said compound is 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid 4-chloro-benzylamide.

25. The compound of claim 1, wherein:

R₂is hydrogen;

R₃is hydrogen;

R₄is optionally substituted with C_1-6Bicyclic [2.2.1] alkyl]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

R₆is a taskPhenyl optionally substituted with 1-3 substituents independently selected from the group consisting of: halogen, hydroxy, C optionally substituted by halogen_1-20Alkoxy radical, C_1-20Alkyl, optionally substituted with halogen, cyano or C_1-20Phenoxy radical of alkyl and optionally substituted with C_1-20Alkyl or halogen phenyl.

26. The compound of claim 25, wherein the compound is:

2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -benzyl) -1, 1-dioxo-1. lamda⁶-isothiazolidine-3-carboxylic acid ((1R,2R,3R,4S) -3-isopropyl-bicyclo [2.2.1]Hept-2-yl) -amide; or

27. The compound of claim 1, wherein:

R₂is dimethylamino;

R₃is hydrogen;

R₄is2, 6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl;

R₅is2, 2-dimethylpropyl; and

28. The compound of claim 27, wherein said compound is 2- (3- { [ (6-chloro-2-fluoro-3-methoxy-benzyl) - ((S) -1-dimethylaminomethyl-3, 3-dimethyl-butyl) -amino]-methyl } -5-dimethylamino-benzyl) -1, 1-dioxo-1. lamda.⁶-isothiazolidine-3-carboxylic acid ((1S,2S,3S,5R) -2,6, 6-trimethyl-bicyclo [3.1.1]Hept-3-yl) -amide hydrochloride.

29. The compound of claim 1, wherein:

R₂is hydrogen or dimethylamino;

R₃is hydrogen;

R₄is optionally substituted with C_1-6Bicyclic or tricyclic cycloalkyl of alkyl having 7 to 10 ring atoms, or R₄Is 4-chlorobenzyl;

or R₃And R₄Together with the nitrogen atom to which they are attached form 4-phenyl-piperidinyl or 4- (4-chloro-phenyl) -4-hydroxy-piperidinyl;

R₆is composed of

R₈Selected from the group consisting of: bromine; a cyanophenoxy group; a difluoromethoxy group; an ethoxy group; fluorine; a hydroxyl group; iodine; a methoxy group; phenyl optionally substituted with one substituent being halogen or C_1-6An alkyl group; and a phenoxy group optionally substituted with one substituent selected from the group consisting of: halogen, cyano and C_1-6An alkyl group;

or R₈And R₉Together are-O-CH₂-O-；

R₁₀Is hydrogen or methyl;

R₁₁selected from the group consisting of: chlorine; fluorine; hydrogen; isopropyl group; and a methyl group;

30. The compound of claim 29, wherein:

R₂is hydrogen;

or R₈And R₉Together are-O-CH₂-O-; and

31. The compound of claim 29, wherein:

R₈selected from the group consisting of: bromine; a difluoromethoxy group; an ethoxy group; fluorine; iodine; phenyl substituted with one substituent being halogen or C_1-6An alkyl group; and a phenoxy group optionally substituted with one substituent selected from the group consisting of: halogen, cyano and C_1-6An alkyl group;

or R₈And R₉Together are-O-CH₂-O-；

R₁₀Is hydrogen; and

R₁₁is hydrogen.

32. The compound of claim 29, wherein R₆Selected from the group consisting of:

2-bromo-6-hydroxy-3-methoxyphenyl;

6-chloro-2-fluoro-3-methoxyphenyl;

6-chloro-2-fluoro-3-methylphenyl;

5-chloro-2-hydroxyphenyl;

2- (4-cyanophenoxy) -phenyl;

2, 6-difluoro-3-bromophenyl;

2, 3-difluoro-6-methoxyphenyl;

2-difluoromethoxyphenyl;

2-ethoxyphenyl;

4' -fluoro-biphenyl-2-yl;

2-fluoro-6-phenoxyphenyl;

2- (4-fluorophenoxy) -phenyl;

2-hydroxy-4-methylphenyl;

2-hydroxy-5-methylphenyl;

2-iodophenyl;

5-isopropyl-2-methoxyphenyl;

2' -methyl-biphenyl-2-yl;

2, 3-methylenedioxyphenyl;

2- (4-methylphenoxy) -phenyl; and

2,3, 6-trifluorophenyl group.

33. A composition comprising a therapeutically acceptable amount of a compound of claim 1 and a carrier.

34. A unit dosage formulation comprising a therapeutically acceptable amount of a compound of claim 1 and a carrier.

35. A process for the preparation of a compound according to any one of claims 1 to 32, comprising the steps of

(i) Reacting a compound of formula 2

Conversion to a compound of formula 3

(ii) Reacting a compound of formula 3 to form a compound of formula 4

(iii) Condensing a compound of formula 4 with a compound of formula 5

To give a compound of formula 6

(v) Converting the compound of formula 7 to the compound of formula 8

(vi) Condensing a compound of formula 8 with a compound of formula 9

To give a compound of formula 10

(vii) Converting the compound of formula 10 to the compound of formula 1

Wherein

R₁Is C_1-6Alkyl radical and

R₂、R₃、R₄、R₅and R₆As defined in claim 1.

36. Use of a compound of any one of claims 1-32 in the manufacture of a medicament for treating or controlling a cell proliferative disorder.

37. Use of a compound according to any one of claims 1 to 32 in the manufacture of a medicament for the treatment or control of breast, colon, lung and prostate tumors.