AU2003262101B2 - Arylpiperazines and their use as metalloproteinase inhibiting agents (MMP) - Google Patents

Description

P/00/011 Regulation 3.2

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION STANDARD PATENT Invention Title: Arylpiperazines and their use as metalloproteinase inhibiting agents (MMP) The following statement is a full description of this invention, including the best method of performing it known to us: ARYLPIPERAZINES AND THEIR USE AS METALLOPROTEINASE INHIBITING AGENTS

(MMP)

The present invention relates to compounds useful in the inhibition of metalloproteinases and in particular to pharmaceutical compositions comprising these, as well as their use.

The compounds of this invention are inhibitors of one or more metalloproteinase enzymes. Metalloproteinases are a superfamily of proteinases (enzymes) whose numbers in recent years have increased dramatically. Based on structural and functional considerations these enzymes have been classified into families and subfamilies as described in N.M. Hooper (1994) FEBS Letters 354:1-6. Examples of metalloproteinases include the matrix metalloproteinases (MMP) such as the collagenases (MMPI, MMP8, MMP13), the gelatinases (MMP2, MMP9), the stromelysins (MMP3, MMIP 10, MMP 11), matrilysin (MMP7), metalloelastase (MMP12), enamelysin (MMP19), the MT-MMPs (MMP14, MMP 15, MlN P_6, 6MM 7);Pthe.reprolysin or adamalysin or MDC family which includes the secretases and sheddases such as TNF converting enzymes (ADAM10 and TACE); the astacin family which include enzymes such as procollagen processing proteinase (PCP); and other metalloprotenases such as aggrecanase, the endothelin converting enzyme family and the angiotensin converting enzyme family.

Metalloproteinases are believed to be important in a plethora of physiological disease processes that involve tissue remodelling such as embryonic development, bone formation and uterine remodelling during menstruation. This is based on the ability of the metalloproteinases to cleave a broad range of matrix substrates such as collagen, proteoglycan and fibronectin.

Metalloproteinases are also believed to be important in the processing, or secretion, of biological important cell mediators, such as tumour necrosis factor (TNF); and the post translational proteolysis processing, or shedding, of biologically important membrane proteins, such as the low affinity IgE receptor CD23 (for a more complete list see N. M.

Hooper et al., (1997) Biochem J. 321:265-279).

Metalloproteinases have been associated with many disease conditions. Inhibition of the activity of one or more metalloproteinases may well be of benefitin these disease conditions, for example: various inflammatory and allergic diseases such as, inflammation of the joint (especially rheumatoid arthritis, osteoarthritis and gout), inflammation of the gastrointestinal tract (especially inflammatory bowel disease, ulcerative colitis and gastritis), inflammation of the skin (especially psoriasis, eczema, dermatitis); in tumour metastasis or invasion; in disease associated with uncontrolled degradation of the extracellular matrix such as osteoarthritis; in bone resorptive disease (such as osteoporosis and Paget's disease)); in diseases associated with aberrant angiogenesis; the enhanced collagen remodelling associated with diabetes, periodontal disease (such as gingivitis), corneal ulceration, ulceration of the skin, post-operative conditions (such as colonic anastomosis) and dermal wound healing; demyelinating diseases of the central and peripheial nervous systems (such as multiple sclerosis); Alzheimer's disease; and extracellular matrix remodelling observed in cardiovascular diseases such as restenosis and atheroscelerosis.

A number of metalloproteinase inhibitors are known; different classes of compounds may have different degrees of potency and selectivity for inhibiting various metalloproteinases. We have discovered a new class of compounds that are inhibitors of metalloproteinases and are of particular interest in inhibiting MMP-13, as well as MMP-9.

The compounds of this invention have beneficial potency and/or pharmacokinetic properties.

MMP 13, or collagenase 3, was initially cloned from a cDNA library derived from a breast tumour M. P. Freije et al. (1994) Journal of Biological Chemistry 269(24):16766- 16773]. PCR-RNA analysis of RNAs from a wide range of tissues indicated that MMP13 expression was limited to breast carcinomas as it was not found in breast fibroadenomas, normal or resting mammary gland, placenta, liver, ovary, uterus, prostate or parotid gland or in breast cancer cell lines (T47-D, MCF-7 and ZR75-1). Subsequent to this observation MMP13 has been detected in transformed epidermal keratinocytes Johansson et al., (1997) Cell Growth Differ. 8(2:243-250], squamous cell carcinomas Johansson et al., (1997) Am. J. Pathol. 151(2):499-508] and epidermal tumours Airola et al., (1997) J.

Invest. Dermatol. 109(2):225-231]. These results are suggestive that MMP13 is secreted by transformed epithelial cells and may be involved in the extracellular matrix degradation and cell-matrix interaction associated with metastasis especially as observed in invasive breast cancer lesions and in malignant epithelia growth in skin carcinogenesis.

Recent published data implies that MMP13 plays a role in the turnover of other connective tissues. For instance, consistent with MMP13's substrate specificity and preference for degrading type II collagen G. Mitchell et al., (1996) J. Clin. Invest.

97(3):761-768; V. Knauper et al., (1996) The Biochemical Journal 271:1544-1550], MMP13 has been hypothesised to serve a role during primary ossification and skeletal remodelling [M.

Stahle-Backdahl et al., (1997) Lab. Invest. 76(5}:717-728; N. Johansson et al., (1997) Dev.

Dyn. 208(3:387-397], in destructive joint diseases such as rheumatoid and osteo-arthritis

[D.

Wernicke et al., (1996) J. Rheumatol. 23:590-595; P. G. Mitchell et al., (1996) J. Clin.

Invest. 97(3):761-768; 0. Lindy et al., (1997) Arthritis Rheum 40(8): 1391-1399]; and during the aseptic loosening of hip replacements Imai et al., (1998) J. Bone Joint Surg. Br.

80(4):701-710]. MMP13 has also been implicated in chronic adult periodontitis as it has been localised to the epithelium of chronically inflanied mucosa human gingival tissue J. Uitto et al., (1998) Am. J. Pathol 152(6):1489-1499] and in remodelling of the collagenous matrix in chronic wounds Vaalamo et al., (1997) J. Invest. Dermatol. 109(1):96-101].

MMP9 (Gelatinase B; 92kDa TypeIV Collagenase; 92kDa Gelatinase) is a secreted protein which was first purifed, then cloned and sequenced, in 1989 Wilhelm et al (1989) J. Biol Chem. 264 (29) 17213-17221. Pubished erratum in J. Biol Chem. (1990) 265 (36) 22570.). A recent review of MMP9 provides an excellent source for detailed information and references on this protease T.H. Vu Z. Werb (1998) (In Matrix Metalloproteinases. 1998. Edited by W.C. Parks R.P. Mecham. pp 115 148. Academic Press. ISBN 0-12-545090-7). The following points are drawn from that review by T.H. Vu Z. Werb (1998).

The expression of MMP9 is restricted normally to a few cell types, including trophoblasts, osteoclasts, neutrophils and macrophages. However, it's expression can be induced in these same cells and in other cell types by several mediators, including exposure of the cells to growth factors or cytokines. These are the same mediators often implicated in initiating an inflammatory response. As with other secreted MMPs, MMP9 is released as an inactive Pro-enzyme which is subsequently cleaved to form the enzymatically active enzyme.

The proteases required for this activation in vivo are not known. The balance of active MMP9 versus inactive enzyme is further regulated in vivo by interaction with TIMP-I (Tissue Inhibitor of Metalloproteinases a naturally-occurring protein. TIMP- 1 binds to the Cterminal region of MMP9, leading to inhibition of the catalytic domain of MMP9. The balance of induced expression of ProMMP9, cleavage of Pro- to active MMP9 and the presence of TIMP-1 combine to determine the amount of catalytically active MMP9 which is present at a local site. Proteolytically active MMP9 attacks substrates which include gelatin, elastin, and native Type IV and Type V collagens; it has no activity against native Type I collagen, proteoglycans or laminins.

There has been a growing body of data implicating roles for MMP9 in various physiological and pathological processes. Physiological roles include the invasion of embryonic trophoblasts through the uterine epithelium in the early stages of embryonic implantation; some role in the growth and development of bones; and migration of inflammatory cells from the vasculature into tissues. Increased MMP9 expression has observed in certain pathological conditions, therebye implicating MMP9 in disease processed such as arthritis, tumour metastasis, Alzheimer's, Multiple Sclerosis, arid plaque rupture in atherosclerosis leading to acute coronary conditions such as Myocardial Infarction.

In a first aspect of the invention we provide compounds of the formula I R3 n O R1 R2 wherein ring B is a monocyclic or bicyclic alkyl, aryl, aralkyl, heteroaryl or heteroaralkyl ring comprising up to 12 ring atoms and containing one or more heteroatoms independently chosen from N, O, and S; alternatively ring B may be biphenyl; ring B may optionally be linked to ring A by a C1-4.alkyl or a C1-4 alkoxy chain linking the 2-position of ring B with a carbon atom alpha to X2; each R3 is independently selected from hydrogen, halogen, N02, COOR wherein R is hydrogen or C1-6alkyl, CN, CF3, C1-6 alkyl, -S-C1-6 alkyl, -SO-C 1-6 alkyl, -S02-C1-6 alkyl ,C1-6 alkoxy and up to C10 aryloxy, n is 1,2 or 3; P is -(CH 2 wherein n 0, 1, 2, or P is an alkene or alkyne chain of up to six carbon atoms; where X2 is C, P may be -Het-, -(CH[R6])n-Het-, -Het-(CH[R6]n-or -Het-(CH[R6])n-Het-, wherein Het is selected from CO-, SO-, -S02-, -NR6-, or -0wherein n is 1 or 2, or P may be selected from -S02-N(R6)- and N(R6)-S02-, and R6 is hydrogen, C1-6 alkylup to C10 aralkyl or up to C9 heteroalkyl; Ring A is a 5-7 membered aliphatic ring and may optionally be mono- or disubstituted by optionally substituted C1-6 alkyl or C1-6 alkoxy, each substituent being independently selected from halogen, C1-6 alkyl or an oxo group; XI and X2 are independently selected from N and C, where a ring substituent on ring A is an oxo group this is preferably adjacent a ring nitrogen atom; Y is selected from -S02- and -CO-; Z is -CONHOH, Y is -CO- and Q is selected from -C(R6)(R7)-CH2-, and -N(R6)-CH2- wherein R6 is as defined above, and solely in relation to Q as here defined, R6 may also represent up to C10 aryl and up to C9 heteroaryl, and R7 is H, C1-6 alkyl, or together with R6 forms a carbocyclic or heterocyclic spiro 5, 6 or 7 membered ring, the latter containing at least one heteroatom selected from N, 0, and S; Z is -CONHOH, Y is -S02- and Q is selected from and -C(R6)(R7)- CH2-; or Z is -N(OH)CHO and Q is selected from -CH(R6)-,-CH(R6)-CH2-, and -N(R6)- CH2-; R1 is H, Cl-6 alkyl, C5-7 cycloalkyl, up to ClOaryl, up to C Oheteroaryl, up to C12aralkyl, or up to C12heteroarylalkyl, all optionally substituted by up to three groups independently selected from N02, CF3, halogen, C1-4alkyl, carboxy(CI-4)alkyl, up to C6cycloalkyl,-OR4, -SR4, C1-4alkyl substituted with -OR4, SR4 (and its oxidised analogues), NR4, N-Y-R4, or C1-4alkyl-Y-NR4, with the proviso that where Ri is -OH, -OR4, -SR4, or NR4, or N-Y-R4 then Z is not -N(OH)CHO, or R1 is 2 ,3,4,5, 6 -pentafluorophenyl; R4 is hydrogen, C1-6 alkyl, up to C10. aryl or up to C10 heteroaryl or up to C9 aralkyl, each independently optionally substituted by halogen, N02, CN, CF3, CI-6 alkyl, -S- C -6 alkyl, -SO-C -6 alkyl, -S02-C1-6 alkyl or C1-6 alkoxy; R2 is H, C 1-6 alkyl, or together with RI forms a carbocyclic or heterocyclic spiro 5, 6 or 7 membered ring, the latter containing at least one heteroatom selected from N, 0, and S; also the group Q can be linked to either RI or R2 to form a 5, 6 or 7 membered alkyl or heteroalkyl ring comprising one or more of O, S and N.

Any alkyl groups outlined above may be straight chain or branched.

Convenient values for the above groups include the following: ring A= a 5-6 membered aliphatic ring, such as a piperazine ring, and may optionally be mono- or di-substituted by optionally substituted C1-6 alkyl or C1-6 alkoxy, each substituent being independently selected from halogen, C 1-6 alkyl or an oxo group; R3 hydrogen, halogen, N02, CF3, C1-4 alkyl, and Cl-4 alkoxy, n is 1 or 2, such as individually 4-fluoro, CF3, 4-chloro and 3,4-dichloro;.

ring B =monocyclic or bicyclic aryl, aralkyl or heteroaryl having up to 10 ring atoms, especially monocyclic aryl, aralkyl or heteroaryl having up to 7 ring atoms, more especially monocyclic aryl or heteroaryl having up to 6 ring atoms, such as a phenyl or pyridyl ring; P -(CH2)n- wherein n is 0 or 1, or or -CO-N(R6)-, one or both of X2and X =N,or X1 is Nor X2isC Y -S02-, Y= -CO-, Q -CH(R6)-CH2-, and -N(R6)-CH2- wherein R6 is hydrogen or C1-6 alkyl; also where Q is linked to RI or R2 to form a C5-7 alkcyl or heteroalkcyl ring such as a cyclohexyl ring; RI hydrogen, CI -6alkyI, C5-7 cycloalkyl, up to C12aralkyI, up to C I Iheteroarylalkyl, up to CI10 aryl or heteroa-yl such as up to C6 aryl; all optionally substituted by up to three halogen atoms, or by CF3;- R2 =hydrogen, or together with RI represent a carbocyclic or heterocyclic spiro 6 membered ring, such as a tetrahydropyran ring; R4 up to CIO aryl optionally substituted by halogen, N02, CN, CF3, CI-6 alkyl, -S- CI1-6 alkyl, -SO-C 1-6 alkyl, -S02-C 1-6 alkyl or Cl1-6 alkoxy; Z =-CONIIGH-, Z -N(OH)CHO.

Preferred values for the above groups include the following: R3 hydrogen, halogen such as chlorine, bromine or fluorine, N02, CF3, methyl, ethyl, methoxy, ethoxy, particularly methoxy or fluorine;, ring B phenyl, pyridyl and pyrimidyl, more especially phenyl, 2-pyridyl and 2,4-pyrimidyl; both X2 and X I are N; Y -S02-; Q =-CH2-; RI is phenyl, 4-trifluoromethylphenyl, phenethyl, phenpropyl, isobutyl, cyclopentyl, benzyloxymethyl, 3 ,4-dichlorophenyl, pyridyl, pyridylethyl, thiophenylpropyl, bromothiophenyl, pyrimidinylethyl, pyrimidinylpropyl, pyridylethyl, pyridylpropyl or together with R2 is spirocyclohexane or spiro-4-pyran; R2 is hydrogen Z -N(OH)CHO.

More preferred values include R3 being halogen, the substituent is preferably meta or para to the ring junction, where ring B is phenyl then especially 4-fluoro and where ring B is pyridyl then or 4-chloro (as appropriate); Q -CH2-.

Preferred combinations of Rings B and A include phenyl and piperazinyl; pyridyl and piperazinyl, and pyrimidine and piperazinyl respectively.

Particular alicyclic, fused and heterocyclic rings for ring B include any one of C N~ NU N X> Nv rvC N^ N^

N

hL

S

P

S

N

NJ

N

N

N

T-N

IN

K

N

N

N

N

D

vS

P

0

S

N"

N-N

S

Particular rings for ring A include any one of N

-C-

-Cir- -N N-- -N N- \-i

-N

NC

and its corresponding seven membered analogue(s).

It will be appreciated that the particular substitituents and number of substituents on rings A and B are selected so as to avoid sterically undesirable combinations. This also applies to rings as may be formed by R1 and Q, R2 and Q as well as R6 and R7.

Where optically active centres exist in the compounds of formula I, we disclose all individual optically active forms and combinations of these as individual specific embodiments of the invention, as well as their corresponding racemates.

Specific compounds include M/Z M+ I (ESP+) 43 8 0 0 M/Z M+ I (ESP+) 455 MIZ M+I (ESP+) 487 0

K

MIZ M+1 (ESP+) 454

NI'S

N.

M/Z M+1 (ESP+) 420 M/Z M+1 (ESP+) 451 0 I0 t 11 NI 0

S

F/\O

NN-S

0 N o -N M/Z M±I (ESP+) 457 M/Z M+1 (iESP+) 438 rN "ICI M/Z M+lI (ESP+) 496 N 1- N MIZ M+ I (ESP+) 471 -11- F 0 cI F ~N N-S 0 MIZ M+1 (ESP+) 528 CI N N-S N 0- b N Br MIZM+1 (ESP+) 511 0

N

NN C M/Z M+I (ESP+) 475 0 Fl clN N-SF 0 VZ M+1 (ESP+) 495 -12cl N

N

0 0s N'IZ M+ I (ESP+) 455 .MIZ M+ I (ESP+) 468 0 cl \N N-S' N 0 MIZ M±1 (ESP+) 455 cl0 M/Z M-f- (ESP+) 427 CA 0 0

N-

NL\N

N-S%

11 0 0 MIZ M±1 (ESP+) 456 -13- 0 0-N 0 cl I )N

N/

-N

N

0 I IN cI N N-S0

N-

N 0 oN 0-N 0 0 0 clN

N-S-

N I

N

0 C1

C

l N N-S FN N N So..N 0 F 0 HO o

R

wherein R phenyl or phenethyl and O O F N

NOH

OH

R

wherein R isobutyl or a spiro-4-pyran ring As previously outlined the compounds of the invention are metalloproteinase inhibitors, in particular they are inhibitors of MMP13. Each of the above indications for the compounds of the formula I represents an independent and particular embodiment of the invention. Whilst we do not wish to be bound by theoretical considerations, the compounds of the invention are believed to show selective inhibition for any one of the above indications relative to any MMP1 inhibitory activity, by way of non-limiting example they may show 100- 1000 fold selectivity over any MMP 1 inhibitory activity.

In addition we have found that compounds of the formula 1 wherein ring B is phenyl, pyridyl (such as 2-pyridyl or 3-pyridyl, especially 2-pyridyl) ring optionally mono- or disubstituted, preferably mono-substituted, by halogen (for example chlorine P is a direct bond; ring A is a piperazinyl or piperidinyl:ring, Y is -S02- and Q is C1-4alkylene (for example especially -CH2-; R1 is as defined for Formula 1 and is especially 2phenyipropyl, 2 -(2-pyridyl)propyl, 2-(3-pyridy)propyl, 2 -(4-pyridyl)propyl, phenyl, benzyloxymethyl, 4-phenylbutyl, 2-phenylbutyl, or 2-(2-thienyl)propyl; and Z is N(OH)CHO; are of particular use as aggrecanase inhibitors ie. inhibitors of aggrecan degradation. Of particular note are compounds of the formula I wherein ring B is a phenyl, 3methylphenyl, 4-fluorophenyl, 3-chlorophenyl, 4-chlorophenyl, or 3,4-dichlorophenyl ring or 5-chloro-2-pyridyl; P is a direct bond; ring A is piperidinyl or piperazinyl especially -16piperazinyl, Y is S02, Q is -CH2-, Z is -N(OH)CHO and RI is phenyl, phenbutylene, phenisopropylene, 2-pyridylethylene, 2-pyridyfisopropylene, 3-pyridylisopropylene, 4pyridylisopropylene, or 4-chlorophenyloxydimethylmethylene. Also of mention are compounds of the formula I wherein ring B is phenyl monosubstituted by chlorine or fluorine, especially 4-chiorophenyl and 4-fluorophenyl; P is a direct bond; ring A is piperidinyl, Y is S02, Q is Z is -CONHOH and RI is hydrogen, i-butyl, or spiro-tetrahydropyranyl.

Particular compounds include

B--P

4-F-Ph 3-Cl-Ph 4-F-Ph 4-F-Ph 4-F-Ph 3-Cl-Ph 3-CH3-Ph 4-F-Ph 5-CI-2-Pyridyl 4-F-Ph 4-F-Ph 4-F-Ph 4-Br-Ph 4-F-Ph 4-F-Ph

A

PIP

PIP

PIP

pip Piperidinyl pip

PIP

pip pip

PIP

PIP

PIP

Piperidinyl

PIP

PIP

PIP

Y

S02 S02 S02 S02 S02 S02 S02 S02 S02 S02 S02 S02 S02 S02 S02 S02

Q

0H2 CH2 C H2 01-2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 0H2 R1 CH2CH(CH3)Ph PhCH2CH2CH2CH2 PhCH2OCH2 4-Pyrdy1CH(CH3)CH2 PhCH(CH3)CH2 (R)-2-PhCH(CH3)CH2 3-Pyddy1CH(CH3)CH2 Ph CH2CH(CH2CH3)Ph 3-PyridyiCH(CH3)CH2 2-thiophenyCH-(CH3)CH2 2-CH3PhCH2CH2 3-PyridyCH(CH3)CH2 PhCH(CH3)CH2 4-F-PhCH(CH3)CH2 2-PyrazinylCH(CH3)CH2 wherein PIP =piperazinyl RH reverse hydroxamate group and R2 hydrogen The compounds of the invention may be provided as pharmaceutically acceptable salts. These include acid addition salts such a.-hydro chloride, hydrobromide, citrate and maleate salts and salts formed with phosphoric and sulphuric acid.. In another aspect suitable salts are ba se salts such as an alkali metal salt for example sodium or potassium, an alkaline earth metal salt for example calcium or magnesium, or organic amine salt for example.

triethylamine.

-17- They may also be provided as in vivo hydrolysable esters. These are pharmaceutically acceptable esters that hydrolyse in the human body to produce the parent compound. Such esters can be identified by administering, for example intravenously to a test animal, the compound under test and subsequently examining the test animal's body fluids. Suitable in vivo hydrolysable esters for carboxy include methoxymethyl and for hydroxy include formyl.

and acetyl, especially acetyl.

In order to use a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof for the therapeutic treatment (including prophylactic treatment) of mammals including humans, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.

Therefore in another aspect the present invention provides a pharmaceutical composition which comprises a compound of the formula or a pharmaceutically acceptable salt or an in vivo hydrolysable ester and pharmaceutically acceptable carrier.

The pharmaceutical compositions of this invention may be administered in standard manner for the disease condition that it is desired to treat, for example by oral, topical, parenteral, buccal, nasal, vaginal or rectal adminstration or by inhalation. For these purposes the compounds of this invention may be formulated by means known in the art into the form of, for example, tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely divided powders or aerosols for inhalation, and for parenteral use (including intravenous, intramuscular or infusion) sterile aqueous or oily solutions or suspensions or sterile emulsions.

In addition to the compounds of the present invention the pharmaceutical composition of this invention may also contain, or be co-administered (simultaneously or sequentially) with, one or more pharmacological agents of value in treating one or more disease conditions referred to hereinabove.

The pharmaceutical compositions of this invention will normally be administered to humans so that, for example, a daily dose of 0.5 to 75 mg/kg body weight (and preferably of to 30 mg/kg body weight) is received. ThTs daily dose may be given in divided doses as necessary, the precise amount of the compound received and the route of administration depending on the weight, age and sex of the patient being treated and on the particular disease condition being treated according to principles known in the art.

-18- Typically unit dosage forms will contain about 1 mg to 500 mg of a compound of this invention.

Therefore in a further aspect, the present invention provides a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof for use in a method of therapeutic treatment of the human or animal body.

In yet a further aspect the present invention provides a method of treating a metalloproteinase mediated disease condition which comprises administering to a warmblooded animal a therapeutically effective amount of a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof.

In another aspect the present invention provides a process for preparing a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof which process comprises a) reacting a compound of the formula (II) or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof with a compound of the formula (III) B

A

P- X 2 1

II

R3 n Y

ZI

O il R1 R2 wherein Xi' represents X or a precursor of X (whether by modification or displacement) or an activated form of X suitable for reaction with Y:; Yi represents Y, a precursor of Y, or an activated form of Y suitable for reaction with

XI

1 by way of non-limiting example, when X is C then Xi may be derivatised to include a precursor of Y for reaction with a compound of formula III wherein Y' is a precursor of Y; Z' represents a protected form of Z, a precursor of Z (whether by modification or displacement of Z) or an activated form of Z; -19and where Q -(CH 2 then by reacting a compound of the formula IX with an appropriate compound of the formula R1-CO-R2 to yield an alkene of the formula X, which is then converted to a compound of the formula XI wherein Z* is a hydroxylamine precursor of the group Z, and then converting Z* to the group Z, all as set out below:

R

6 B

RI

P- X" P 2 X SO 2

CH

2

IX

R3R B

A

R3 R2 B X6 R R 2 XI or b) reacting a compound of the formula (IV) or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof with a compound of the formula P' X X-

Z-

Y Q -IV R1 R2 B

V

R3, wherein B' represents a suitable ring function or substituent group for reaction with P'; Z' is as hereinbefore defined; and P' represents a suitably activated form of the linker P for reaction with B' or where X2 N then P1 may be present on ring A rather than ring B or, as required, the linker P may be formed by appropriate reaction of precursor groups P" and provided on rings B' and A respectively, or vice versa.

A compound of the formula (II) is conveniently prepared by reacting a compound of the formula (VI) with a compound of the formula (VII)

BI

VI

R3 n X 2 VII wherein B' represents a suitable ring function or substituent group, X2 represents X or a precursor of X (whether by modification or displacement) or an activated form of X suitable for reaction with B' and wherein B' and X21 when reacted together provide the linker P between ring A and ring B in the compound of formula By way of non-limiting example, when X 2 is N then ring B is suitably derivatised to introduce the linker P via and when X 2 is C then both ring B and ring A are suitably derivatised to provide the linker P by the reaction of B' and X2

I

-21- It wkill be appreciated that many of the relevant starting materials are commercially available. In addition the followxing table shows details of aldehyde intermediates and their corresponding registry numbers in Chemical Abstracts.

RCHO Chemical Abstracts Registr-y Numbers 2-methy1-2- 4-chlorophenoxy ropionaldehyde 6390-87-0 2 -methyl-2(4-chiorophenylthio)-propionaldehyde 56421-90-0 4 -phenoxybutyraldehyde 19790-62-6 cyclhexvacealdeyde5664-21-1 3-cyclohexylpropionaldehyde 4361-28-8 4 -cyclohexylbutyadhe 180-41-9 3 3 -py1dylbu-yrnaldhyde 79240-21-4 3 2 -pyridyl)propionaldehyde 2057-32-1 eraldehyde368-83 3 -phenyl-4-methylvaleraidehyde 54784-84-8 3- 2 -yrazinyl)butyraldehyde 177615-94-0 furan-2-carboxaldehvde 221525-60-6 3-( 4 -chlorophenyl)propionaldehvde 75677-02-0 3 4 -fluorophe-,Nvl-propzonaldhde I63416-70-6 3 4 -*pvridv1)propionaldchvde 120690-80-4 4 -phenylbutraldehyde 170650-98-3 2 -pv1nidv~carboxa~dehvde 1121-60-4 3 -(3-pyridyl)propionadehvde -T1802-16-0 3 2 -furv1)propionadchvde 4543-51-5 4 2 -pvridyl)b-utvraldehvde 90943-32-1 4-Bromothiophene-2 carboxaldhvde 189 71-75-8 cyclo pentanecarboxaldehyde 87-3-7 Benzoxazole. I-pipcrazinvl)-(9C 1) 111628-39-8 Benzothiazole. 2 -piperazinYl)-(9C 1) 55745-83-0 Benzoxazole 5-choro-2-( 1-pipez-zinv1)-(9C 1) 140233-44-9 Benotiaole6-hlro--( -iperazinvl)-(9C 1) 153025-29-7

L

3 -pvridvl-5-bromo-carboxa;dehyde I _7113118-81-3 Aldehydes without Chemical Abstracts Reistrv Numbers 3 -(2-pyrimidyl) propionaldehyde. To a solution of 2 -Bromopyrimidine (7.95 g, 0.05 M) in acetonitrile (150 m.L) was added propargylalcohol (4.2 g, 0.075 M bis- (triphenylphosphine)-palladium(I 1)chloride (750 mg, 1 copper iodide (100 mg, 0. mM) and triethylamine (25mL, 0.25 M) and the mixture was stirred and heated at 70 0 C for 2 hours. An additional amount of propargyl alcohol 1&g 0.03 8 bis-(triphenylphosphine)- -22palladium(l 1)chloride (375 mg, 0.5 mil), and copper iodide (50 mg, 0.25 mil) was then added to the reaction mixture which was stirred and heated at 70 0 C for an additional 1 hour.

The reaction mixture was evaporated to dryness and the residue which was preadsorbed on to silica, chromatographed. Elution with ethyl acetate gave 3-(2-pyrimidyl) prop-2-yn-3-ol as a yellow solid 4.45 g (66 NMR (CDC13) 2.9 (1H, 4.5 (2H, 7.3.( 1H, 8.8 2H, MS found MN- 135 3 -(2-pyrimidyl) prop-2-yn-l-ol (4.45 g, 0.033 M) was dissolved in ethyl acetate (140 mL), 10 Pd/C (890 mg) was added and the mixture stirred under an atmosphere of hydrogen for 6 hours. The reaction mixture was filtered through Celite and the filtrate evaporated to give 3-(2-pyrimidyl) propan-1-ol as a yellow oil, 4.15 g (91 NMR (CDCI,) 2.1 2H, 3.2 (2H, 3.8 2H, 7.2 1H, 8.7 2H, d) MS found MH' 139.

3-(2-pyrimidyl) propan- -ol was oxidized to give 3-(2-pyrimidyl) propionaldehyde as a yellow oil NMR (CDC1 3 3.0 2H, 3.4 2H, 7.1 IH, 8.7 (2H, 9.9 1H, s) using the Swern oxidation described in this patent.

Using the procedure described above the following aldehydes were prepared 4-(2-pyrimidyl) butyraldehyde by using 3-butyn- -ol in place of propargylalcohol NMR CDC13 9.8(1H, 8.6 (2H, 7.15 (1H, 3.0 (2H, 2.5 (2H, 2.2 (2H, m).

butyraldehyde by using 3-butyn-l-ol in place of propargylalcohol and in place of 2-bromopyrimidine NMR CDCI 3 9.8 (1H, 9.1 (1H, 8.6 2h, 2.7 (2H, 2.55 (2H, 2.0(2H, m).

4 -(2-pyridyl) butyraldehyde by using 3-butyn-l-ol in place of propargylalcohol and 2bromopyridine in place of 2-bromopyrimidine NMR CDCI 3 9.8 (1H, 8.6 (lH, 7.6 1H, 7.1( 2H m 2.8 (2H, 2.55 (2H, t), 2.0(2H, m).

The compounds of the invention may be evaluated for example in the following assays: Isolated Enzyme Assays Matrix Metalloproteinase family including for example MVMP13.

Recombinant human proMMP 13 may be expressed and purified as described by Knauper et a. Knauper et al., (1996) The Biochemical Journal 271:1544-1550 (1996)].

The purified enzyme can be used to monitor inhibitors of activity as follows: purified -23proMMP 13 is activated using 1mM amino phenyl mercuric acid (APMA), 20 hours at 21 0

C;

the activated MIMP13 (11.25ng per assay) is incubated for 4-5 hours at 35'C in assay buffer 1M Tris-HCI, pH 7.5 containing 0. 1M NaCI, 20mM CaCI 2 0.02 mM ZnCI and 0.05% Brij 35 using the synthetic substrate 7 -methoxycoumarin-4yl)acetyl.Pro.Leu.Gly.Leu.N-3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl.Ala.Arg.NH2 in the presence or absence of inhibitors. Activity is determined by measuring the fluorescence at lex 3 28nm and kem 393nm. Percent inhibition is calculated as follows: Inhibition is equal to the [Fluorescenceplus inhibitor FluorescenCebackground] divided by the [Fluorescenceius inhibitor Fluorescencebackground] A similar protocol can be used for other expressed and purified pro MMPs using substrates and buffers conditions optimal for the particular MMP, for instance as described in C. Graham Knight et al., (1992) FEBS Lett. 296(3):263-266.

Adamalvsin famil includin2 for exam le TNF cnvertase The ability of the compounds to inhibit proTNFo convertase enzyme may be assessed using a partially purified, isolated enzyme assay, the enzyme being obtained from the membranes of THP-I as described by K. M. Mohier et al, (1994) Nature 370:218-220. The purified enzyme activity and inhibition thereof is determined by incubating the partially purified enzyme in the presence or absence of test compounds using the substrate 4',5'-Dimethoxy-fluoresceinyl Ser.Pro.Leu.Ala. Gln.Ala. Val.Arg. Ser. Ser. Ser.Arg. Cys(4-(3succinimid -yl)-fluorescein)-NH2 in assay buffer (50mM Tris HCI, pH 7.4 containing 0.1% Triton X- 100 and 2mM CaCI 2 at 26'C for I 8 hours. The amount of inhibition is determined as for MIMPI3 except Xex 4 90nm and Xem 530nm were used. The substrate was synthesised as follows. The peptidic part of the substrate was assembled on Fmoc-NH-Rink- MBHA-polystyrene resin either manually or on an automated peptide synthesiser by standard methods involving the use of Fmoc-amino acids and O-benzotriazolI tetramethyluronium hexafluorophosphate (HBTU) as coupling agent with at least a 4- or fold excess of Fmoc-amino acid and HBTU. Seri and Pro 2 were double-coupled. The following side chain protection strategy was employed; Ser'(Bu'), Gln 5 (Trityl), Arg 8 ,1 2 (Pmc or Pbf), Sero" 0 "'(Trityl), Cys (Trityl). Following assembly, the N-terminal Fmoc-protecting group was removed by treating the Fmoc-peptidyl-resin with in DMF. The amino-peptidylresin so obtained was acylated by treatment for 1.5-2hr at 70 0 C with 1.5-2 equivalents of -24- 4',5'-dimethoxy-fluorescein-4(5)-carboxylic acid [Khanna Ullman, (1980) Anal Biochem.

108:156-161) which had been preactivated with diisopropylcarbodiimide and I-hydroxybenzotriazole in DMF]. The dimethoxyfluoresceinyl-peptide was then simultaneously deprotected and cleaved from the resin by treatment with trifluoroacetic acid containing 5% each of water and triethylsilane. The dimethoxyfluoresceinyl-peptide was isolated by evaporation, trituration with diethyl ether and filtration. The isolated peptide was reacted with 4 -(N-maleimido)-fluorescein in DMF containing diisopropylethylamine, the product purified by RP-HPLC and finally isolated by freeze-drying from aqueous acetic acid.

The product was characterised by MALDI-TOF MS and amino acid analysis.

Natural Substrates The activity of the compounds of the invention as inhibitors of aggrecan degradation may be assayed using methods for example based on the disclosures of E. C. Arner et al., (1998) Osteoarthritis and Cartilage 6:214-228; (1999) Journal of Biological Chemistry, 274 6594-6601 and the antibodies described therein. The potency of compounds to act as inhibitors against collagenases can be determined as described by T. Cawston and A. Barrett (1979) Anal. Biochem. 99:340-345.

Inhibition of metalloproteinase activity in cell/tissue based activity Test as an agent to inhibit membrane sheddases such as TNF convertase The ability of the compounds of this invention to inhibit the cellular processing of TNFa production may be assessed in THP-1 cells using an ELISA to detect released TNF essentially as described K. M.'Mohler etal., (1994) Nature 370:218-220. In a similar fashion the processing or shedding of other membrane molecules such as those described in N. M.

Hooper et al., (1997) Biochem. J. 321:265-279 may be tested using appropriate cell lines and with suitable antibodies to detect the shed-protein.

Test as an agent to inhibit cell based invasign The ability of the compound of this invention to inhibit the migration of cells in an invasion assay may be determined as described in A. Albini et al., (1987) Cancer Research 47:3239-3245.

Test as an agent to inhibit whole blood TNF sheddase activity The ability of the compounds of this invention to inhibit TNFcx production is assessed in a human whole blood assay where LPS is used to stimulate the release of TNFa.

Heparinized (10Units/ml) human blood obtained from volunteers is diluted 1:5 with medium (RPMI1640 bicarbonate, penicillin, streptomycin and glutamine) and incubated (160ul) with 20 l of test compound (triplicates), in DMSO or appropriate vehicle, for 30 min at 37 0

C

in a humidified (5%C02/95%air) incubator, prior to addition of 20ul LPS coli. 0111:B4; final concentration 10pg/ml). Each assay includes controls of diluted blood incubated with medium alone (6 wells/plate) or a known TNFa inhibitor as standard. The plates are then incubated for 6 hours at 37°C (humidified incubator), centrifuged 2 0 00rpm for 10 min; plasma harvested (50-100) and stored in 96 well plates at -70 0 C before subsequent analysis for TNFa concentration by ELISA.

Test as an agent to inhibit in vitro cartilage degradation The ability of the compounds of this invention to inhibit the degradation of the aggrecan or collagen components of cartilage can be assessed essentially as described by K.

M. Bottomley et al., (1997) Biochem J. 323:483-488.

Pharmacodynamic test To evaluate the clearance properties.and bioavailability of the compounds of this invention an ex vivo pharmacodynamic test is employed which utilises the synthetic substrate assays above or alternatively HPLC or Mass spectrometric analysis. This is a generic test which can be used to estimate the clearance rate of compounds across a range of species.

Animals rats, marmosets) are dosed iv or po with a soluble formulation of compound (such as 2 0%w/v DMSO, 60% w/v PEG400) and at subsequent time points 5, 15, 120, 240, 480, 720, 1220 mins) the blood samples are taken from an appropriate vessel into 10U heparin. Plasma fractions are obtained following centrifugation and the plasma proteins precipitated with acetonitrile (80%w/v final concentration). After 30 mins at the plasma proteins are sedimented by centrifugation and the supernatant fraction is evaporated to dryness using a Savant speed vac. The sediment is reconstituted in assay buffer and subsequently analysed for compound content using the synthetic substrate assay. Briefly, a compound concentration-response curve is constructed for the compound undergoing -26evaluation. Serial dilutions of the reconstituted plasma extracts are assessed for activity and the amount of compound present in the original plasma sample is calculated using the concentration-response curve taking into account the total plasma dilution factor.

In vivo assessment Test as an anti-TNF agent The ability of the compounds of this invention as ex vivo TNFo inhibitors is assessed in the rat. Briefly, groups of male Wistar Alderley Park (AP) rats (180-210g) are dosed with compound (6 rats) or drug vehicle (10 rats) by the appropriate route e.g. peroral intraperitoneal subcutaneous Ninety minutes later rats are sacrificed using a rising concentration of CO 2 and bled out via the posterior vena cavae into 5 Units of sodium heparin/ml blood. Blood samples are immediately placed on ice and centrifuged at 2000 rpm for 10 min at 4 0 C and the harvested plasmas frozen at -20 0 C for subsequent assay of their effect on TNFc production by LPS-stimulated human blood. The rat plasma samples are thawed and 1751l of each sample are added to a set format pattern in a 96U well plate. Fifty .1 of heparinized human blood is then added to each well, mixed and the plate is incubated for min at 37 0 C (humidified incubator). LPS (251il; final concentrationl0 Og/ml) is added to the wells and incubation continued for a further 5.5 hours. Control wells are incubated with of medium alone. Plates are then centrifuged for 10 min at 2000 rpm and 200pl of the supernatants are transferred to a 96 well plate and frozen at -20 0 C for subsequent analysis of TNF concentration by ELISA.

Data analysis by dedicated software calculates for each compound/dose: Percent inhibition of TNFc= Mean TNFa (Controls) Mean TNFac (Treated) X 100 Mean TNFc (Controls) Test as an anti-arthritic agent Activity of a compound as an anti-arthritic is tested in the collagen-induced arthritis (CIA) as defined by D. E. Trentham et al., (1977) J. Exp. Med. 146,:857. In this model acid soluble native type II collagen causes polyarthritis in rats when administered in Freunds incomplete adjuvant. Similar conditions can be used to induce arthritis in mice and primates.

-27- Test as an anti-cancer agent Activity of a compound as an anti-cancer agent may be assessed essentially as described in I. J. Fidler (1978) Methods in Cancer Research 15:399-43 9, using for example the B 16 cell line (described in B. Hibner et al., Abstract 283 p 7 5 10th NCI-EORTC Symposium, Amsterdam June 16 19 1998).

The invention will now be illustrated but not limited by the following Examples:

EXAMPLES

Example 1 1 2 -(N-formvl-N-hvdroxvamino)-2-phenviethanesulfonvl-4-(4fluorophenyl)pierazine F N N-S HO'N

H

0 To a solution of 1-[2-(hydroxyamino)-2-phenylethanesulfonyl]-4-(4fluorophenyl)piperazine (338 mg, 0.89 mmol) in THF (5 ml) and formic acid (2 ml) was added a preformed mixture of formic acid (2 ml) and acetic anhydride (0.5 ml). The mixture was stirred at room temperature for one hour. The mixture was evaporated in vacuo and toluene (2 x 5 ml) was added and evaporated in vacuo. The residue was taken in CH 2 C2 methanol (6 ml, 9 1) and silica (1 g) was added. The mixture was stirred for 18 hours. The silica was filtered off and rinsed with CH 2 C2 methanol (9 The residue was purified on silica gel (eluant: CH 2 C2 MeOH 4 to give the title compound as a light orange solid (220 mg, 61 'H NMR (CDC1 3 8.45 and 8.15'(s, 1H), 7.39 5H), 6.97 2H), 6.88 2H), 5.89 and 5.35 1H), 4.05 and 3.85 1H), 3.30-3.53 5H), 3.20-3.10 4H); MS (ESI): 408 (MHI), 430 (MNa); EA: calculated for CgH22FN 3 0 4 S: C 56.01, H 5.44, N 10.31, S 7.87, Found: C 56.01, H 5.52, N 10.04, S 7.39.

The starting material was prepared as follows: i) To a solution of 1-(4-fluorophenyl)piperazine (35 g, 194 mmol) and pyridine (17.5 ml) in dry dichloromethane (200 ml) at 0°C was added methanesulfonyl chloride (20 mi, 258 mmol) dropwise. The mixture was stirred for 3 hours at room temperature. The mixture was -28washed with water and extracted with dichloromethane (2 x 100 ml). The organic layers were dried with MgSO 4 and evaporated in vacuo. The residue was triturated and washed with methanol to give l-( 4 -fluoropheiyl)-4-(methanesulfonyl)piperazine (39.35 g) as white crystals.

'H I'lvR (CDC1 3 7.00 (in, 2H1), 6.90 (in, 2M1, 3.40 (mn, 4H), 3.20 (in, 41-1), 2.83 3H).

ii) To a solution of LDA [8.5 inmol prepared by slow addition of n-butyl lithium ml, 8.5 inmol, 2.5 M in hexane) to a solution of diisopropvlamine (860 mng, 8.5 mmol) in dry THY (5 ml) at -78"C] at -78*C was added a solution of 1-(4-fluorophenyl)-4- (methanesulfonyl)piperazine (I g, 3.87 mmol) in THE (25 ml) dropwise. The mixture was .stir-red at -78'C for 1 hour and a solution of diethylchiorophosphate (670 mg 3.87 inmol) in THE (3 ml) was added. The mixture was sti rred at -78'C for I hour and benzaldehyde (450 mng; 4.24 inmol) in THE (3 ml) was added, The mixture was gently warmed to room temperature and stirred for 18 hours. The mixture was washed with aqueous arnmonium chloride and extracted with ethyl acetate. The organic layers were washed with water, brine and dried over MgSO 4 Purification of the residue on silica (eluant dichloromethane) afforded 1 4 -fluorophenyl)-4(trans-3-styrenesulfonyl)piperazine as a white powder (621 mg, 46%.

'H NUR (CDCl 3 7.50 (in, 311), 7.43 (mn, 3M1, 6.97 (mn, 2H), 6.89 (in, 2H), 6.71 111, J= 15.4 Hz), 3.37 (in, 411), 3.19 (in, 4H).

iii) To a solution of I 4 -fluorophenyl)-4-(trans- 3-styrenesulfony1)piperazine (620 mg, 1.79 mmol) in THE (20 ml) was added hydroxylainine (3 ml, 50 aqueous solution). The mixture was stirred for 18 hours. The solvent was evaporated. The residue was dissolved in dichioromethane and washed with water. The organic layer was dried on MgSO 4 to give (hydroxyamino)-2-phenylethanesulfonylp4-(4-fluorophenyl,)piperaine (730 mng), 'H NN{R (CDC] 3 7.4-7.1 (mn, 511), 6.97 (in, 211), 6.87 (mn, 2H), 5.95 (s br, 1H1), 4.74 111, 4.60 (dd, III, J= 4 Hz, Y=8 8 Hz), 3.56 (dd, 1H, J= 8.8 Hz, 14.3 Hz), 3.40 (in, 4H), 3.19 (dd, lH, J= 4 Hz, F1= 14.3 Hz), 3.12 (in, 411).

Exaqmple 2 Si milarly the following compounds were obtained: Comp~ound Data 0 Ph F N .N

H

HO'

0 .N H HO Ny 0 C1 NN-S0 b C1 HO'

H

0 F N \/N-S6, H'N yH 0 MS (EST): 436 458 (MNa") MS (ESI): 400 (MHi), 422 MS (ESI): 476 (MI-f, 35 C1) 498 (MNa, 35 Cl) MS (ESI): 422 (MNa') Example 3 P

H,

F- N/ N- N OH To a solution of N-(2,4-dirnethoxybenzyloxy)-N-(2,4,6-trimethoxybenzyl)-3 fluorophenyl)piperazine- I sulfonyl]propionamide (125 mg, 0. 19 minol) in dichloromethane (2 ml) was added triethylsilane (66 )Al, 0.42 rnmol) and trifluoroacetic acid (150 pl). The mnixture was stirred at room temperature for 4 hours. The solvents were evaporated in vacuo. The residue was purified by chromatography on silica (eluant dichloromethane, then ethyl acetate then dichloromethane -10 MeOH) to give 35 mg of the title compound.

'H NMR (DMSO d-6 CF 3 COOD): 7.16 4H), 3.36 6H), 3.25 4H), 2.45 2H, J 7.4 Hz); MS (ESI): 332 (MN 4 354 (MNa').

The starting material was obtained as follows: i) A solution of 3-mercaptopropionic acid (20 g, 185 mmol) in acetic acid (150 ml) water (30 ml) at 0° C was reacted with gaseous chlorine (preferably condensed at -78° C, 20 ml). After chlorine had distilled, the solvents were evaporated in vacuo; toluene was added and evaporated to give 1,2-oxathiolane-5-one 2-dioxyde (36.12 g).

'H NMR (DMSO 2.70 2H, J= 7.2 Hz), 2.50 2H, J= 7.2 Hz).

ii) A solution of 1,2-oxathiolane-5-one 2-dioxide (3.8 g, 28 mmol) in thionyl chloride ml) and DMF (5 drops) was stirred at room temperature for 18 hours. The mixture was heated at 40 0 C for 1 hour. The solvents were evaporated; toluene was added and evaporated in vacuo to give crude 3-chlorosulfonylpropionyl chloride (NMR purity 70 3.58 g).

'H NMR (CDC1 3 4.02 2H, J= 7.2 Hz), 3.63 2H, J= 7.2 Hz) iii) To a solution of 3 -chlorosulfonylpropionyl chloride (500 mg, 1.83 mmol, 70 purity) and diisopropylethylamine (75 ul) in dichloromethane (5 ml) at -78 0 C was added a solution of O-dimethoxybenzyl-N-trimethoxybenzylhydroxylamineRn 1 (664 mg 1.83 mmol) and diisopropylethylamine (320 gl, 1.83 mmol) in dichloromethane (5 ml) dropwise over 2 hours. After 30 minutes, a solution of 1-( 4 -fluorophenyl)piperazine (330 mg, 1.83 mmol) and diisopropylethylamine (320 .l 1.83 mmol) in dichloromethane (5 ml) was added to the reaction mixture. The solution was warmed to room temperature and stirred for 2 hours. The solution was partitioned between dichloromethane and IN hydrochloric acid. The organic layers were washed with brine and dried over MgSO 4 Chromatography of the residue on silica gel (eluant ethyl acetate petroleum ether: gradient from 50/50 to 80/20) gave N-(2,4dimethoxybenzyloxy)-N-(2,4,6-trimethoxybenzyl)-3-[ 4 4 -fluorophenyl)piperazine-1sulfonyl]propionamide (260 mg).

MS 661 -31- Example 4 N-hvdroxy-3-f4-benzvlrinerazine-l-sulfonvllp~ronionamide '0 H -N

-O

00 In a manner analogous to that described in Example 3, from 4-benzylpiperazine and 3chiorosulfonyipropionyl chloride there was obtained the title compound.

'H NMVR(DMSO d-6 CF 3 COOD)-: 7.50 (mn, 5H), 4.41 2H), 3.78 (in, 3.41 (mn, 4$, 3.18 (mn, 2H), 2.43 211, J= 7.1 Hz); MS (ESI) 3 28 (Ivflfl.

Example N-hvdroxv,-3-r4-(4-fluorophenvF)niperazine-1 -sulfonvll-2-isobt-vli~rotpionamide.

F ~N

NNSNOH

0 0 To a solution of N-(2,4-dimethoxybenzyloxy)-3 -[4-(4-fluorophenyl)piperazine- 1 sulfonyl]-2-isobutylpropionamide (220 mg) in dichloromethane (4 ml) was added trifluoroacetic acid (200 gl) and triethylsilane (145 pA). The mixture was stirred at room temperature for 15 minutes, evaporated in vacuo and the residue was purified on silica gel (eluant: dich-loroinethane-ether-inethanol (8020:0.5) to dichloroinethane-inethanol (80:20) to give the title compound (88 ing, 56%) 'H NMR (DMSO 10.72 1H), 7.08 (in, 2H), 6.99 (in, 2H), 3.37 (dd, 1K J= 8.4 Hz, F= 14.3 Hz), 3.27 (mn, 4H), 3.15 (mn, 47H, 3.00 (dd, Ili, J=4 Hz, JY= 14.3 Hz), 2.62 (in, 1.6-1.2 (in, 3H), 0.89 3H, J =6.6 Hz), 0.85 31-1, J= 6.6 Hz); MS (ESI): 388 (MHl 4 410 (MNa).

The starting material was obtained as follows: -32i) A solution of 3 -acetylthio-2-isobutylpropionic acid [obtained by Michael addition of thiolacetic acid onto 2 -isobutylacrylic acid] (7 g, 34.3 mmol), benzyl bromide (4.29 ml, 36 mmol) and DBU (5.2 ml, 35 mmol) in toluene (55 ml) was stirred for 18 hours at room temperature. The solvents were evaporated in vacuo. The residue was partitioned between ethyl acetate and 5% sodium bicarbonate. The organic layer was washed with brine and dried over MgSO 4 Purification of the residue by chromatography on silica gel (eluant: dichloromethane-ether gave benzyl 3 -acetylthio-2-isobutylpropionate (8.4 g) MS (ESI) 317 (MINa).

ii) A solution of benzyl 3 -acetylthio-2-isobutylpropionate (588 mg, 2 mmol) in acetic acid (12 ml) water (1.6 ml) at 0 0 C was reacted with gaseous chloride (prealably condensed at -78 0 C, 1.9 ml). After chlorine had distilled, the solvents are evaporated in vacuo to give crude benzyl 3-chlorosulfonyl-2-isobutylpropionate (630 mg).

MS 318 iii) A solution of benzyl 3 -chlorosulfonyl-2-isobutylpropionate (630 mg, 2 mmol), 1- 4 -fluorobenzyl)piperazine (378 mg, 2.1 mmol) and triethylamine (340 4l, 2.4 mmol) in dichloromethane (15 ml) was stirred at 00 C for 18 hours. After evaporation of the solvents, the residue was partitioned between ethyl acetate and water. The organic layer was washed with brine and dried over MgSO 4 After evaporation of the solvent in vacuo, the residue was purified by chromatography on silica gel (eluant: dichloromethane ether to give benzyl 3 4 4 -fluorophenyl)piperazine- 1 -sulfonyll-2-isobutylpropionate (640 mg) MS (EI) 462 iv) A solution of benzyl 3-[ 4 4 -fluorophenyl)piperazine- I1-sulfonyl]-2isobutylpropionate (630 mg) in methanol (10 mi) was hydrogenated under 40 PSI pressure for 18 hours in the presence of palladium on charcoal (63 mg, The catalyst was removed by filtration and the solvents were removed in vacuo to give fluorophenyl)piperazine- 1 -sulfonyll-2-isobutylpropionic acid (460 mg).

MS (ESI): 373 (MNI), 395 (MNa').

v) To a solution of 3-[ 4 -(4-fluorophenyl)piperazine- I -sulfonyl]-2-isobutylpropionic acid (230 mg, 0.62 mmol), 2 4 -dimethoxybenzylhydroxylamine Rc"I (124 mg, 0.68 mmol), DMAP (75 mg, 0.62 mmol) in DMF (1 mi) was added N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (152 mg, 0.8 mmol). The mixture was stirred at room temperature for 2 days. The reaction mixture was poured in water and extracted with ethyl -33acetate. The organic layer was washed with 5% sodium bicarbonate, brine and dried over MgSO 4 Purification of the residue on silica gel (eluant: dichioromethane ether: gradient from 9/1 to 8/2) gave N-(2,4-dimethoxybezyloxy)-3-[4-(4-fluorophenyl)piperazine-lsulfonyl]-2-isobutylpropionamide (158 mng).

'H NMIR (CDCI 3 :8.21 1K), 7.30 (mn, 1KH), 6.97 (mn, 2H), 6.88 (mn, 2H), 6.46 (in4 2H), 4.95 211), 3.82 6H), 3.5 0 (dd, I1K, J= 9 Hz, JF= 14.2 Hz), 3.3 7 (in, 4H), 3.14 (mn, 4H), 2.84 (dd, IH, 3= 14.2 Hz, 2 Hz), 2.60 IK), 1.7-1.2 (mn, 3H), 0.90 (in, 6K).

Example 6 4-f4-(4-fluoronhenvl)piuerazine- I-sulfonvlmethvlltetrihvdropvran-4-(N-hvdroxv carboxamide) 0

H

"n 0 To a solution of 4-[4-(4-fluorophenyl)piperazine-lI-sulfonylmethyljtetrahydropyran-4carboxylic acid (470 mng, 1.21 minol) in dichioromethane (S ml) was added oxalyl chloride (700 mg, 5.6 mmol) and DM/F (18 gl). The mixture was heated at 3 50 C for 1 hour. After evaporation of the solvents, the crude acid chloride dissolved in dichioromethane (4 ml) was added to a ice-cooled solution of hydroxylamine (440 p1, 50 aqueous solution) in TE (8 ml). The mixture was stirred for 90 minutes at. room temperature. After evaporation of the solvents, the residue was triturated in dichloromethane-ether-inethanol (80: 20 The resulting solid was washed with water and ethyl acetate and dried to give the title compound as white crystals (230 mg, 47%) 'H NMP. (DMSO 10.56 (s Or, 1M), 8.74 (s br, 1H), 7.07 (in, 2K), 6.99 (in, 2H), 3.66 (in, 2H), 3.47 (in, 2K), 3.40 2H), 3125 (in, 4H), 3.16 (in, 4H), 1.99 (in, 2K), 1.72 (in, 2K); MS (ESI): 402 424 (MNa).

The starting material was prepared as follows: Thiolacetic acid (760 gl, 10 minI) and tributylphosphine (2.5 ml, 10 inmol) in DMF ml) was added dropwise to a ice-cooled suspension of sodium hydride (530 mg, 60 in oil, 13 mmcl) in DMF (1.5 ml) under an argon atmosphere. The mixture was stirred at0 0

C

for 30 minutes. To the above solution was added 2,7-dioxaspiro[3,5]nonane-1I-one r~12 (1.4 g, 10 mmol) in DMF (10 ml). The mixture was stirred at 0' C for 3 0 minutes and at room temperature for I8 hours. The reaction mixture was diluted with ether. The precipitate was filtered and dried to give 4-(acetylthiomethyl)tetrahydropyran-4.carboxylic acid sodium salt.

'H NMR (DMSO 3.65-3.40 (mn, 4H), 2.99 2H), 2.27 3H), 1. 86 (mi, 2H), 1.23 (n, 2H).

(ii) Using the same procedure described in Example 5 ii), iii), iv), v) except that no DBU was used in step 1, from 4-(acetylthiomethyl)tetrahydropyran-4-carboxylic acid sodium salt was obtained 4-[4-(4-fluorophenyl)piperazine- I -sulfonylinethyl]tetrahydropyran-4carboxylic acid (490 mng).

4-(acetyithiomethyl)tetrahydropyran-4-( carboxylic acid benzyl ester): MS (ESD): 331 4-(chloro sulfonylrnethyl)tetrahydropyran-4-( carboxylic acid benzyl ester): MS (ESI): 3 54 (MINa'); 4-[4-(4-fluorophenyl)pipera-zine- I -sulfonylmethyl]tetrahydropyran-4carboxylic acid benzyl ester: MS 477 499 fluo rophenyl)piperazine- I -sulfonylm ethyl] tetrahydropyran-4-carboxyl I c acid: MS (ESI): 387 (Iv1Hl), 409 (MINa').

Ref 1; B.Barlaam, A.Hamon, M.Maudet; Tetrahedron Lett, 1998, 39 7865 Ref 2: F. Hoffmann-La Roche, Agouron Pharm.; Eur. Patent Appi. EP 7803 )86.

Example 7 1- 12-(N-formvl-N-hvdroxvamino)-2-phenvlethzinesulfonilj-4-phenvlpinerazine 0%S /0 0 N N

NK

To a solution of 1 2 -(hydroxyamino)-2-phenylethanesulfonyl]-phenylpiper-azine (140 mg) in TKF (0.75 ml) and formic acid (0.25 ml) was added a preformed mixture of formic acid (0.58 ml) and acetic anhydride (0.29 ml). The solution was stirred at ambient temperature for 18 hours. The mixture was evaporated in vacuo, diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate solution, dried (Na 2

SO

4 and evaporated. The residue was purified by chromatography eluting with 1 methanol in dichloromethane to give l-phenyl-(4- trans-b-styrenesulfonyl)piperazine (420 mg) as a foam (105 mg).

'H NMR (d6-DMSO at 373K): 9.60 1H), 8.25 1H), 7.40 2H), 7.30 (m, 3H), 7.20 2H), 6.90 2H), 6.75 1H),'5.60 1H), 3.85 (dd, 1H), 3.60 (dd, 1H), 3.30 4H); 3.15 4H) m/z: 390 The starting material was prepared as follows: A solution of phenylpiperazine (487 mg) in dichloromethane (6 ml) containing triethylamine (0.63 ml) was added dropwise over 5 minutes to trans-b-styrenesulfonyl chloride (638 mg) in dichloromethane (4 ml). The solution was stirred at ambient temperature for 18 hours. The solution was diluted with dichloromethane and washed with water, dried (Na 2

SO

4 and evaporated. The residue was purified by chromatography eluting with 1 methanol in dichloromethane to give I-phenyl-(4- trans-bstyrenesulfonyl)piperazine (420 mg) as a solid.

'HNMR (d6-DMSO): 7.75 2H), 7.40 4H), 7.30 1H), 7.20 (dd, 2H), 6.90 2H), 6.80 (dd, 1H), 3.20 8H) m/z 329 To a solution of 1-phenyl-4-( trans-b-styrenesulfonyl)piperazine (108 mg) in THF (3 ml) was added hydroxylamine (0.45 ml, 50 aqueous solution). The mixture was stirred at ambient temperature for 18 hours. Solvent was removed in vauo and the residue dissolved in dichloromethane, washed with water, dried (Na 2

SO

4 and evaporated to give the product 1-[2- (hydroxyamino)-2-phenylethanesulfonyl]-4-phenylpiperazine as a foam (120 mg).

'H NMR (d6-DMSO): 7.50 1H), 7.40 2H), 7.30 5H), 6.90 2H), 6.80 (dd, 1H), 5.90 1H), 4.20 1H), 3.60 (dd, 1H), 3.40 (dd, 1H), 3.20 4H); 3.10 (m, 4H) m/z 362 -36- Example 8 1-t2-(N-formvl-N-hvdroxamino)-2-(cuinoline-4lethane.1.sulfonvl14(4.

fluorophenvl~piperazine 1\ P N N N-N-S, 0 To a suspension of I 2 -(N-hydroxyamirio)-2-(quinoline-4-yl)ethane- I -sulfonyl]-4-(4fiuorophenyl)piperazine (148 mg, 0.34 mmol) in TH-F (2 ml) CH 2 C 2 (2 ml) was added methyl-3-formyl-l,3,4-thiadiazole-2(3H)thione( 1 (140 mg, 0.87 inmol). The mixture was stirred for 3 h. After addition of methanol (2 ml) and silica (I the mixture was stirred for 18 h. The solids were filtered. The filtrates were washed with sat. NaHCO 3 and brine.

Evaporation of the solvents followed by trituration in acetonitrile CH 2 C1 2 gave the starting material (60 mg). Chromatography of the mother liquors with acetonitrile CH 2 Cl 2 1) gave the title compound (20 ing, 13%).

1 H-NMiR (CDC1 3 8.97 (in, IH), 8.21 (in, 2H), 8.01 IH), 7.8-7.65 (in, 3H), 6.97 (mn, 2H), 6.86 (mn, 5.66 (mn, 3.55-3.1 (in, IOM); MS 459 (MI-I).

The starting material was prepared from quinoline-4-carboxaldehyde and I1- (fluorophenyl)-4-(inethanesulfonyl)piperazine in a similar manner to Example I ii-iii): 1 88 mg: MS (ESI): 431 HIPLC tR (Column TSKgeI super ODS 2mm 4.6mmn x 5cm, gradient methanol! water 20 to 100% in 5 min, flow rate: 1.4 mI/mn): 3.43 mnn Yazawa, Goto, S. Tetrahedron Lett., 1985, 2 6, 3 703 Exampnle 9 1-fl -(N-forniyl-N-hvdroxvamino)-I -(3,4-dichlororphenvl)nentane-2-sulfonvl-4-(4fluorophenflh~yerazine.

CCi HO

NY

0 Similarly to Example 1, the syn and anti diastereoisomers of 1IEl -(N-hydroxyarnino)l-( 3 4 -dichlorophenyl)pentane-2-sulfonyl]-4-(4-fluorophenyl)piperazine were converted to the title compound (as 2 diastereoisomers): diastereoisomer I from the less polar hydroxylamine: 36 mg, 70%; 'H-NMIR (CDC1 3 8.45 and 8. 10 IH), 7.6-7.2 (in, 3H), 7.0-6.8 (in, 2H), 5.96 arnd 5.18 (in, 1K), 3.8-3.4 (mn, 5H1), 2.9-3.15 (mn, 4H), 2.0-1.0 (4n 4H), 0.88 and 0.76 3H, J 7 Hz); MS (ESI): 540 (M{ 35 C1, 35 542 (M{ 3 7 Cl, 35 Cl})Na-).

diastereoisomer 2 from the more polar hydroxylamine: 49 mg, 63%; 'H-NNM (CDC1 3 8.28 and 8.13 lH), 7.6-7.2 (mn, 3H), 7.0-6.85 (mn, 5.54 and 5.02 (in, lH), 3.45-3.9 SF1), 3.15 (in, 4H), 1.7-1.2 (mn, 4H), 0.76 3H, J 7 Hz); MS 540

I

5 Cl, 35 Cl 542 37 C1, 35 C1 }Nai):- The starting material was prepared as follows: Similarly to Example 1 from I -(4-fluoropheny)piperazine and 1 -butanesulfonyl chloride was obtained 1-(4-fluorophenyl)-4-(butane-lI-sulfonyl)piperazine 84 similarly to Example 1 ii), this was reacted with 3,47dichlorobenizaldehyde to give 4-(4-fluorophenyl)- 3 ,4-dichlorophenyl)pent-1-ene-2-sulfonyl]piperazine as a mixture of Z/E isomers (330 ing, MS (IESI): 457 (M(3 5 Cl, 35 459 (M{ 37 C1, 35 similarly to Example I iii) except that the mixture was refluxed for 37days, this was converted to hydroxyainino)- 1-(3 ,4-dichlorophenyl)pentane-2-sulfonyl]-4-(4-fluorophenyl)pip erazine as the syn and anti diastereoisomers.

Less polar isomer (50 ing, 15%) (TLC: eluant EtOAc CH 2

CI

2 petroleum ether 45-50); 1 H-NMiR (CDCI 3 7.53 1KL J= 2.2 Hz), 7.46 1K, J= 7.4 Hz), 7.27 (in, 1K), -38- 6.97 (in, 2H), 6.88 (mn, 211), 4.63 (mn, IH), 3.55 (mn, 4H), 3.16 (in, 5H), 1.75 (mn, 2H), 1.4 (in, iH), 1.2 (mn, 1ff), 0.77 3K J= 7.4 Hz).

More polar isomer (76 mg, 23 'H-NMIR (CDCI 3 7.52 1K, J= 2 Hz), 7.45 (d, 1KL J= 8 Hz), 7.27 (in, 1I), 6.99 (mn, 2K), 6.89 (n,4 2H), 4.42 (in, 1K), 3.55 (in, 4K), 3.41 (mn, 1K), 3.14 4K), 1.6 (in, 2K), 1.25 (in, 2K), 0.76 3K J=7.3 Hz).

Example Trans 1 -[2-(N-formvl-N-hvdroxvaim ino~cvclohexan -sulfonvll-4-(4-fl uororhenvl)perazine.

0 F-&T N-S

HO-N

CHO

Sim-iarly to Example 1, from trans I -[2-(N-hydroxyamino)cyclohexane- I1-sulfonyl]-4- .(4-fluorophenyl)piperazine was obtained the title compound (68 mg, 23%).

'K-NMv~R (CDCl 3 8.3 9 and 8.02 I1K), 6.98 (in, 2H), 6.8 8 (in, 2K), 4.40 and 3.92 1K), 3.35-3.55 (n,4 5H), 3.15 (in, 4K) 2.35 (mn, 1K), 2.0-1.8 (in, 3H), 1.2-1.6 (mn, 4H); MS (ESI): 408 (MNa The starting material was obtained as follows: i) To a solution of LDA (51 mmol, prepared by slow addition of n-butyl lithium (20.4 ml, 2.5M in hexane, 5I iniol) to solution of diisopropylamine (5.16 g, 51 inmol) in THEI (30 ml) at -78'C) at -78'C was added a solution of 1 -(4-fluorophenyl)-4-(inethanesulfonyl)piperazine (6 g, 23.2 iniol) in THlE (150 ml). The mixture was stirred for I h at -78'C. A solution of 5-chiorovaleryl chloride (4 g, 25.8 iniol) in THYf (20 mnl) was added dropwise.

The mixture was stirred at -78'C for 1 h and at room temperature for 18 h. The solution was diluted with EtOAc and washed with sat. NH4CI and brine and dried over MgSO 4 Chromatography of the residue on silica gel (eluant EtOAc CH 2 Cl 2 petroleum ether (15:35:50)) afforded I -(6-chloro-2-hexanone-lI-sulfonyl)-4-(4-fluorophenyl)piperazine (5.22 g, 60%) as white crystals: MS (ESI): 399 (MNa').

ii) A mixture of this compound (5.22 g, 13.9 iniol) and Nal (42 g) in acetone (90 ml) was refluxed for 5 h. After cooling and partitioning between EtOAc and water, the organic -39layer was washed with 10 NaHSO 3 and brine, and dried over MgSO 4 to give l-(6-iodo-2hexanone- I -sulfonyl)-4-(4-fluoropheny)piperazijne (6.13 g, quantitative) as yellowish crystals: 'H-NMvR (CDC1 3 6.98 (mn, 2M), 6.88 (in, 2H), 4.00 2H), 3.46 41, 3= 4.8 Hz), 3.19 2K, J= 6.6 Hz), 3.16 4K J= 4.8 Hz), 2.79 21-L J= 6.6 Hz), 1.85 (in, 2H), 1.74 2H).

iii) A mixture of this compound (1.27 g, 4.85 minol) and cesium carbonate (8 g, 24.5 inmol) in CH2Cl 2 (90 ml) was stirred at roomf~tmnperature for 4 h. To the mixture was slowly added water and 2N HCI until pH 7. The mixture was extracted with CH 2

CI

2 The organic layer was dried over MgSO 4 Chromatography on silica gel (eluant: EtOAc petroleum ether afforded l-(cyclohexanone-2-sulfonyl>4(4-fluorophenyl)piperazine (880 mg, 53%).

'H-NMR (CDCI 3 6.97 (in, 2H), 6.88 (in, 2K), 3.83 (mn, 1H), 3.48 (in, 4H), 3.12 4H), 2.81 (in, 111), 2.54 (in, IH), 2.46 (in, IH), 2.2-2.0 (mn, 3H), 1.75 (in, 2H); MS (ESI): 363 Further elution (EtOAc petroleum ether afforded 1 -[(tetrahydropyran-2-.

yl)methylidenesufonyl-4(4fluoropheny)piperzne (63 0 mg, 3 8 'H-NMR (CDCI 3 6.98 (in, 2FD, 6.87 (mn, 2H), 5.21 1K), 4.14 2K J= 5.2 Hz), 3.32 (mn, 4M), 3.15 (in, 4H), 2.35 2H, J= 6.6 Hiz), 1.82 (in, 4H); MS (ESI): 363 (MNa 4 iv) To a solution of l-(cyclohexanone-2-sulfonyl)4(4fluorophenyl)piperaine (284 mng, 0. 83 mmol) in methanol-THF (16 ml, 3: 1) at 0 0 C was added sodium borohydride (3.7 ing, 1 minol). The mixture was stirred at 0 0 C for 30 min and at room temperature for 1 h The solvents were evaporated. Saturated NI{ 4 C1 and water were added. The precipitate was filtered, washed with water and dried to give 1 -(2-cyclohexanol- 1 -sulfonyl)-4-(4fluorophenyl)piperazine (250 ing, MS (ESI): 343 (MmH).

v) To a solution of 1 -(2-cyclohexanol- I -sulfonyl)-4-(4-fluoropheny)piperazine (3 mg, 0. 9 mmol) in THE (15 ml) was added triphenylphosphine 18 g 4.5 nunol) and DEAD (712 g.1, 4.5 mmol) dropwise. The mixture was stirred at room temperature for 18 h.

Evaporation of the solvents and purification on silica gel (eluant: EtOAc -petroleum ether, gradient from 2:8 to 3:7) gave 1 -cyclohexene-lI-sulfonyl]L 4 -(4-fluorophenyl)piperazine (285 mg, MS (ESI): 325 vi) Similarly to Example I iii) except that the reaction was heated at 65'C for 30 h, from 1-Iccoeeelsloy)4(-loohnlpprzn (280 mg, 0.86 mg) was obtained trans l-[ 2 -(N-hydroxyanino)cyclohexane I -sulfonyl]- 4 -(4-fluorophenyl)pip erazine (270 mng, 'H-NMiR (CDC 3 6.98 (in, 2H), 6.88 (in, 2H), 3.54 (mn, 4H), 3.34 (in, 2K), 3.14 (mn, 4H), 2.30 (in, 1K), 2.17 (mn, 1K), 2.05 (in, 1H), 1.9-1.2 (mn, 5H); MS (ESD: 358

(MW).

Example 11 Cis 1-T2-(N-formvl-N-hvdroxvamino)cvclohexane-l-sulfonvll-4-(4-fluoronheny).

virierazine.

F-0 Nl N-S

HO-N

CHO

Similarly to Example 1, from cis 1 -[2-(N-hydroxyamino)cyclohexcane- I -sulfonyll-4- (4-fluorophenyl)piperazine was obtained the title compound (18 in-, 'H-NMR (CDC1 3 8.39 and 8.07 1H), 6.98 2H), 6.88 (mn, 211), 4.77 and 4.25 (in, 1H), 3.48 (mn, 3.13 (mn, 2.25-1.3 (mn, MS (EST): 408 (Ma).

The starting material was obtained as follows: 1) A mixture of 1 -(cyclohexanone-2-sulfonyl)-4-(4-fluorophenyl)pperazine (50 mg, 0.14 inmol), hydroxylamine hydrochloride (51 mg, 0.73 minol) and potassium acetate (72 mg, 0.73 minol) in methanol (5 ml) was heated at 70'C for 4 h. The solvents were evaporated.

After partitioning between EtOAc and water, the organic layer was washed with brine and dried over MgSO 4 to give 1 -[2-(N-hydroxyiinino)cyclohexane- 1 -sulfonyl]-4-(4fluorophenyl)piperazine as a white solid (48 mng, MS (ESD: 356 ii) To this compound (210 mg, 0. 6 inmol) in a mixture of TRFf acetic acid (7 mH, 1: 1) was added sodium cyanoborohydride (276 mng, 4.4 iniol). The mixture was stirred at room temperature for 18 h. Water was added and the pH was adjusted to 9. The mixture was extracted with EtOAc. The organic layer was washed with brine and dried over MgSO 4 Chromatography on silica (eluant: EtOAc petroleum ether, gradient from 1: 1 to 8:2) afforded cis I -[2-(N-hydroxyami no)cyclohexane- I -sulfonyl]-4-(4-fluorophenyl)piperazine (97 -41mg, 'H-NTM (CDC1 3 6.98 (in, 6.89 (in, 2H), 3.63 (in, 3.52 (mn, 4H), 3.24 (dt, 111 Jd= 10.6 Hz, 3.5 Hz), 3.15 (mn, 4H), 2.2-1.2 (in, 8H1); MS (ESI): 358 OAR).

Example 12 The following compounds were made using the method outlined in Example 1:

M-H

R2 hydrogen PIP piperazinyl RH reverse hydroxamate A carboxylic acid Mp Mp M+HI Low HihB A Y Q RI 117 117 492-494 4-F-Ph PIP SO2tCH2 2-(5-Br-thiophene). RH 128 128 409 4-F-Ph PIP S021CH2 3-Pyridyl

RH

F125 1125 414 4-F-Ph PIP S021CH2 2-thiophenvi RH 135 1135 414 4-F-Ph PIP S021CH2 3-thiopherrvi RH 409 4-F-Ph PIP S021CH2 2-Pyridvi RH1 ___372 4-F-Ph PIP' CO N (S)-PhCH-2 A __426 14-F-Ph PIP S02 CR2 4-F-Ph RH 14-F-Ph PIP S02 CR2 Gem-di-Me RH __450 4-F-Ph 2-Me-PIP S02 CR2 PhCH2CH2 RH 498* 4-F-Ph PIP S02,CH2 4-CI-PhOCfMe)2 RH 129 130 389 4-Ph Piperidinyl S021CH.2 Ph RH 500-502 3.4-di-Cl-Ph IPIP S02 ICR2 CH2CH(CH3)Ph RH 1466 14-F-Ph IPIP S02 CH2 PhOCH2CH2CH2 RH 1101110 1514* 14-Cl-Ph IPIP 3S02 CR2 4-Cl-PhOC(Me)2 RH 138 140 1550-55213,4-di-Cl-Ph 1PIP. S02 CH2 4-CI-PhOC(Me)2 RHj -42- Mp Mp M+H Low Hig B A Y Q RI Z 69 70 389 Ph 4- S02 CH2 Ph RH 456 4-F-Ph PIP S0d2,lCR2 c-HexvlCH2CH2CH2 1__442 4-F-Ph PIP S02 CR2 lCvciohexvICH2CH2 RH 139 140 1407 4-F-Ph Piperidinvl S02 CR2 lPh RH 172 172 1516 4-F-Ph PIP S02 CR2 14-CI-PhSC(Me)2 RH 517-519 5-C1-2- PEP S02 CR2 14-CI-PhOC(Me)2 RH iPyridyl ___516-518 3-Cl-Ph PIP S02 CH2 4-Cl-PhOC(Me)2 RH 505 4-F-Ph PEP S02 CR2 N-PhCH2-4- RH eridinvi 104 104 548 4-F-Ph PIP S02 CR2 4-Cl-PhS02C(Me)2 RH 135 135 451 4-F-Ph PIP S02 CH2 3- RH Pyridv]CH(CH3)CH2 100 100 1451 4-F-Ph PIP S02 CR2 4- RH _____PyrxdvlCH(CH3)CR2__ 65 451 4-F-Ph PIP S02 CH2 2-yridvlCH(CH3)CH2 RH .69 70 449 4-F-Ph lPiperidinvl S021CH2 PhCH(CH3)CH2 RH 54 55 436 4-F-Ph Piperidinvl S021CH2 2-PvridvlCH2CH2 RH 66 67 449-501 4-F-Phi Piperidinvl S02 CR2 4-C1-PhOC(Me)2 RH 1_ 1480 3-Cl-Ph PIP S02 CH2 jPhCH2CH2CR2CH2 RH 55 1480-482 4-Cl-Ph PIP S02 CH2 j PhCH2CH2CH2CR2 RH 450 4-F-Ph PIp S02 CR-2 RH I PhCH(CH3)CH2 450 4-F-Ph PIP S02 CR2 RH PhCH(CH3)CH2 467 3-Cl-Ph PIP S02 CR2 3- RH PvridvlCH(CH3)CH2 ___464 4-F-Ph PIP S02 CR2 CH2CH(CH2CH3)Ph RH 160 163 428 4-F-Ph PIP 502 CR2 CH-2c-hexvi RH 468 5-CI-2- PIP 502 CH2 3- PH Pvridvi PvridviCH(CH3)CR2 456 4-F-Ph PIP S02 CH2 2- R thiophcnylCH(CH3)C __H2 146 1478 14-F-Ph PIP 502 CR2 PhCH2CH2CH.2CH2 RH 67 68 1450 4-F-Ph PIP S02 CR2 2-CH3PhCH2CH2 RH 76 450 4-F-Ph Pipenidinyl S02 CR2 3- RH RridylCH(CH3)CH2__ 69 70 510-512 4-Br-Ph PIP S02 CR2 PhCH(CH3)CH2 RH 133 135 346 4-F-Ph PIP 502 CH2 CR3 RH 465 4-F-Ph PIP S02 CR2 CH2CH2CH(CH3)3- RH _Pyr 63 .450 4-F-Ph PIP 502 CR2 CH(CH3)CH2Ph RH ___478 4-F-Ph PIP S02 CH2 CH2CHCPri)Ph RH -452 4-F-Ph PEP ]S02 CR2 CR2CH(CH3)Pyrazin RH I_ Yl -43- Mp IMp M+H Low High IB A Y QRI Z 420 2- PIP S02 CR2 PhCH2CH2 RH I Pyrimidinvl.

155 1157 1454 6-CI-4- .PIP S02 CR2 PhCH2CH2 RH Pyrimidinyl______ -452 14-Cl-Ph PIP S02 ICR2 PhCH2CH2 RH -452 3-Cl-Ph PIP S02 CR2 PhCH2CH2 RH -486 3,4-d-CI-Ph PIP S02 CR2 PhCH2CH2 RH 453 5-CI-2- PIP S02 CR2 PhCH2CH2 R Pvridvl 453 3-Cl-2- PIP S02 CR.2 PhCH2CH2 RH Pyridyl 466 4-Cl-Ph Honiapipe S02 CH2 PhCH2CH2 RH razine -419 2-P.Yrdvl PIP S02 ICR2 PhCH2CH2 RH 494 6-CM-- PIP S02 ICR2 3,4-d-Cl-Ph RH Pvrimidinvl 450 6-MeO-4- Pip S02 CH2 PhCH2CHq2 RH Pyrimidinvi 118 120 470 16-C1-4- PIP S02 CR2 PhCI{20CH2 RH Pyrimidinyl 493 6-C1-2- PIP S02 CR2 3,4-di-CI-Ph RH ____Pyridvi 527 5-CF3-2- PEP S02 CR2 3,4-di-CI-Ph RH Pyridvi 562 3-CI-5-CF3- PIP S02 CR2 3,4-d-CI-Ph RH ~2-Pyridyl 469 5-CI-2- PIP S02 CR2 PhCH2OCH2 RH ____Pvridyl___ 493 5-Cl-2- PIP S02 CR2 3,4-di-CI-Ph RH ____Pyridvi 494 6-Cl-4- PIP S02 CR2 4-CF3-Ph RH ____Pvrimidinvi 1523 4-Me-2- PIP S02 C2 3,4-di-Cl-Ph

RH

gquinolvi 468 3-Cl-Ph PIP S02 CR2 PhCH2OCH2 RH 454 2-C1-4- PIP S02 CR2 PhCH2CH2 RH Pyrimidinvi 459 2- Pip S02 CR2 PhCH2CH2 RH Benzoxazol yI 475 2- IPIP S02 CR2 PhCH2CH2 RH Benzthiazol 454 6-CI-3- PER- S02 CR2 PhCH2CH2 RH ____Pvidazinvl 44- NIP Mp jM+H I Low High B A Y Q RI Z ___460 2-Pyridyl PEP S02 [CR2 3,4-di-CI-Ph [RH ___459 2-Pyridyl PIP S02 [CR2 4-CF3-Ph IRH __435 2-Pyridyl PIP S02 [CR2 PhCH20CH2 RH __420 2-Pyvridvl PIP S02 CR2 2-PvridvlCH2CH2 RH 509 7-CI-2- PIP S02 CH2 PhCH2CH2 RH Benzthiazoly ___461 2-Pyrazinyi PIP S02 CR2 .3,4-di-C1-Ph RH __460 2-Pyrazinvl PIP S02 CR2 4-CF3 -Ph RH __436 2-PvrazinvI PIP S02 CR2 PhCH20CH.2 RH ___421 2-Pyrainvl PIP S02 CR2 3-PvridvICH2CH2 RH ___420 2-Pyrazinyl PIP S02 CH2 PhCH2CH2 JRH 470 6-CI-3- PIP S02 CR2 PhCH2OCH2 RH Pyridazinvi 16138 420 4- PIP S02 CR2 [PhCH2CH2 RE 1__421 2-Pyrzinvl PIP S02 CR2 12-PyridviCH2CH2 IRH 417 5-CI-2- PIP S02 CH2 c-Pentyl R ____PyridvI 484 5-CN-2- PIP S02 CH2 3,4-di-Cl-Ph RH Pyri dyi 494 6-CI-2- PIP S02 C2 2-PyridyICH2CR2 R- Benzoxazoly 4-F-Ph PIP S02 CR2 2-Furv! RH 437 4-F-Ph PIP S02 ICR2 3-PvridvlCH2CH2 RH 1437 4-F-Ph PIP S02 ICR2 4-PvridvICH2CR2 RH 42 4-F-Ph PIP S02 ICIR2 PhCH2CH2CH2C

RH

470,472 4-F-Ph PIP S02 JCR2 4-CI-PhCH2CH2 R-H 1450 4-F-Ph PIP S02 CR2 jPhCH2CH2CH2 RI-I 1426 4-F-Ph PIP S02 CH2 ~2-FurvlCH2C-L2 R.H 456 4-F-Ph PIP S02 CR2 2-Thiophenyl- RH I ~CH2CH.2CH2 468 4-F-Ph PIP S02 CH2 4-F- RH I PhCH2CH2CH2 I 1 454 4-F-Ph PIP S02 ICR2 4-F-PhCH2CH2 JRH 1 437 4-F-Ph PIP S02 CR2 2-PvridyICH2CH-2 RH 509,511 5-CI-2- PIP S02 CH2 4-Br-2-Thiophenyl RH 1 420 2-Pyridyl PIP S02 CR2 3-PyridvlCH2CH2 RH 453,455 3-Cl-Ph PIP S02 CR2 3-PvridvlCH2CH.2 RH 1487,489 3.4-di-Cl-Ph PIP S02 CR2 3-PvridylCH2CH2 RH 464 4-F-Ph PIrP SO02 CR2 PhCH2CH2CH2C RH 2 454,456 5-CI-2- PIP S02 CR2 13-PyridylCI-2CH2 RH _Pyndvl I I I_ Mp.- Mp M+H Low High B A IY Q RI Z 466,468 3-Cl-Ph PIP IS02 CR2 PhCH2CH.2CH2 RH 467,469 5-CI-2- PIP S02 CR2 PhCH2CH2CH2 RH Pyridyl 468,470 6-C1-4- PIP IS02 CR2 PhCH2CH2CH.2 RH Pyrixnidinv 455,457 2-C1-4- PIP S02 CR2 3-PyridylCH2CH.2 RH ______Pjrimidinvl 455,457 6-CI-4- PIP -S02 CR2 3-PyridylCH2CH2 RH ____PyrimidinvA 44463-CI-2- PIP S02 CR2 3-PyridylCH2CH2 RH Pyridvl 1__433 2-yrdv PIP S02 CH2 PhCH2CH2CH2 RH 50.3 5-CF3-2- PIP S02 CR2 PhCR2OCH2 RH Pdvl 468,470 P2-C1-4- PIP S02 CR2 PhCH2CH2CH2 RH ____Pyrimidinvi 453,455 3-Cl-Ph PIP S02 JCH2 2-PvridvlCH2CH2 RH 487,489 3,4-di-CI-Ph PIP S2 ]CH2 2-PvridvlCH2CH2 RH 135 137 455,457 6-CI-4- PIP S02 CR2 2-PvridylCH2CH2' RI- Pyrimidinyl 107 109 488 5-CF3-2- PIP IS02 CR2 2-PyridylCR2-CH2 RH Pyriy 451 4-F-Ph PIP IS02 CR2 2- RH _____PvridvlCH2CH2CH2 120 123 452 4-F-Ph PIP S02 CR2 2- RH PyrimidinylCR2 CR2 CR2 452 4-F-Ph PIP S02 CR2 5- RH PyrimidinviCH2CH2 CH2 119 121 468 5-C1-2- PIP S02 CR.2 2- RH- Pvridvl PvridvICH2CH2CH2 469,471 5-CI-2- PIP S02 CR2 5- RH Pyridyl PyrimidinylCH2CH2 CR2 131 134 469,471 5-CI-2- PIP S02 CR2 2- RH Pyridyl PyrimidinylCH2 CR2 426,428 15-CL-2- PIP S02 1 CR2 12-Pyridyl RH _Pyridv1 J MS for C17H24FN305S calcd 402, found 402.

Nf N-S-N 0 0'N 0 -46- The aryl/heteroar-ylpiperazines and piperidines used as starting materials were commercially available or were described in the literature, for example 4-(4-fluorophenyl)-piperidine, CAS number 37656-48-7 Piperazine, 1 '-biphenyl]-4-yl-( 180698-19-5) Piperazine, 1 1,1 '-biphenyl]-3 -yl- (115761-6 Pip erazine, 1 -(2-naphthalenyl)- (57536-91-1) Piperazinone, I -phenyl-(909 17-86-5) 1 H-i ,4-Diazepine, I -(4-chlorophenyl)hexahydro-(4 1885-98-7) Quinoline, 4-methyl-2-( 1-piperazinyl)-( 50693-78-2) Piperazine, 1-(4-phenoxyphenyl)- 62755-61-7 Piperazine, 1 -(3-chlorophenyl)- The 2-methyl-4-(4-fluorophenyl)-piperazine used as starting material was prepared as follows: Sodiurn-t-butoxide ig) was added to a solution of tir-tolylphosphine (0.63 8 g) and palladium acetate (0.3 1 9g in toluene (250 ml) under argon and the mixture was stirred for minutes. 4-Fluoro-bromobenzene (5g) and 2-methylpiperazine (2.85g) were added and the mixture was heated at 1 10 'C for 7 hours, then allowed to cool to ambient temperature and keep at this temperature for 20 hours. The reaction mixture was filtered through Celite®, the filter cake was washed twice with dichloromethane (2X25 mL) and the filtrate was evaporated to dryness. The residue was chrornatographed on silica eluting initially with dichioromethane and then with a mixture of dichlorornethane, methanol and ammonium hydroxide (100: 5: 1) to give 2-rnethyl-4-(4-fluorophenyl)-piperazine, Using this same method and 2,6-dimethylpiperazine as starting material there was obtained 2,6-dimethyl-4-(4-fluorophenyl)-piperazine.

Piperazine, 1 '-biphenyl-4'-fluoro]-4-yl hydrochloride tert-butoxycarbonyl piperazine, 1 I, I'-biphenyl-4'-fluoro] 4 -yl (0.71 2g) was stirred in a mixture of dichloromethane (1 OmI) and trifluoroacetic acid (1 .Oml) for 18 hours at ambient temperature, evaporated in vacuo to a grey solid and used without ifurther purification.

The-tert-butoxycarbonyl piperazine, 1 1,1 '-biphenyl-4'-fluoro]-4-yl used as starting material was prepared as follows: Sodium-t-butoxide (1.3 5g) was added to a solution of bis(diphenylphosphino)- 1,1 V-binapthyl 0 4 6g) and bis(dibenzylideneacetone)palladiurn -47- (0.023g in toluene (25 ml) under argon and then added 4-bromo-4'-fluorobiphenyl (2.51ig) and Il-te rt -butoxycarbonylpiperazine (2.2g) and the mixture was heated at 80'C for 5 hours.

The reaction mixture was filtered, filtrate evaporated in vacuo to a yellow solid which was triturated and then filtered from diethyl ether(20 ml) to give tert-butoxycarbonyl piperazine, 1'-biphenyl-4'-fluoro]-4-yl, 67g), mp 165-166 'C.

NM~R (d6-DMSO) 1.42 9H), 3.15 (in, 4K), 3.48 (in, 4M), 7.02 2K), 7.22 (in, 2K), 7.51 2H), 7.63 (mn, 2KH); m/z 357(M+1).

Example 13 C1 0

C

Acetic anhydride (0.23 ml) was added directly to formnic acid (0.9m1). The solution was stirred at room temperature for 30 minutes and then added a solution of chloropyrimidin-4-yl)tetrahydropyrazin- l-yl] sulfonyl dichlorophenyl)ethyl]hydroxylamine (0.227g) in tetrahydrofuran (5nmi). The solution was stirred at room temperature for 18 hours. The solution was evaporated (water-bath temperature 30 and the residual gum was purified by chromatography. using a I Og silica isolute eluting with CK2Cl2-3% MethanoVCH2Cl2 to give N-[2-([4-(6-chloropyrirnidin-4yl)piperazino]suffonyl I ,4-dichlorophenyl)ethylj-N-hydroxyformnamide 1 78g), 98- 101 0

C.

NMR (d6-DMSO 373 3.31 (in, 4H), 3.70 (dd, 1K), 3.75 (mn, 4M{, 3.95 (dd, 111), 5.61 (vbs, 1K), 6.89 1IM, 7.43 (dd, 1K), 7.60 1K), 7.70 IH), 8.29 111), 8.36 (s, IH);m/z 494

CIC

0 Acetic anhydride (0.63m1) was added directly to formic acid (2.48m1). The solution.

was stirred at room temperature for 30 minutes and then added a solution of chloropyridin-2-yl)piperazino~sulfonyl -3 ,4-dichlorophenyl)ethyl]hydroxylamine (0.61 g) in tetrahydrofuran (1 OmI). The solution was stirred at room temperature for 3 hours.and then diluted with ethyl acetate, neutralised the pH with saturated aqueous sodium hydrogen carbonate solution The ethyl acetate layer was separated, dried (Na2SO4), and evaporated to dryness. The residue was purified by chromatography using a IlOg silica isolute eluting with ethyl acetatelheaxane-80% ethyl acetatelhexane and then evaporated to dryness. The resulting white solid was filtered from diethyl ether to give [4-(5-chloropyridin-2yl)piperazino] sulfonyl)-1I ,4-dichlorophenyl)ethyl]-N-hydroxyformamide (0.43 1 g) 211 212 0

C.

NMvR (d6-DMSO 373 3.30 (in, 4H), 3.80 (mn, 4H), 3.85 (cid, 1K), 3.95 (cid, 1H), 5.58 (vbs, IH), 6.85 (di, 1H), 7.43 (in, IH), 7.58 (in, 2H), 7.85 (d, IH), 8. 10 (ci, 1H), 8.13 IH);in/z 493 N N o~> 0 Acetic anhydride (0.48ml) was added directly to formic acid (1.9in1). The solution was stirred at room temperature for 30 minutes and then added a solution of N-(2-(benzyloxy)-1- ([(4-pyridin-2-ylpiperazino)sulfonyl]methyl }ethyl)hydroxylamine 42g) in tetrahydrofliran The solution was stirred at room temperature for 3 hours.and then diluted with ethyl acetate, neutralised the pH with saturated aqueous sodium hydrogen carbonate solution The ethyl acetate layer was separated, dried (Na2SO4), and evaporated to dryness. The residue was purified by chromatography using a Thbg silica isolute eluting with CH125% Methanol/CH2C12 to give N-(2-(benzyloxy)- 1- {[(4-pyridin-2-ylpiperazino)sulfonyl]methylethyl)-N\-hydroxyforinamide (0.23 3g), 70-75 NMR (d6-DMSO 373 3.25 (dd, 1H), 3.31 (mn, 4H), 3.48 (dd, 1H), 3.65 (in, 3.66 (cid, 1H), 3.70 (dci, 1K), 4.55 (vbs, IH),4.55 2H), 6,70 (mn, 1K), 6.85 (di, 1K), 7.28 (in, 7.32 4H), 7.58 1KH), 8. 17 (in, 2H), 9.45 (bs, 1H);m/z 435 0 Acetic anhydride (0.48m1 was added directly to formic acid (1 .9m1). The solution was stirred at room temperature for 30 minutes and'-then added a solution of N-(3 -pyridin-2-yl- 1 [(4-pyridin-2-ylpiperazino)sulfonyl~methy1propyl)hydroxylamine 15 2g) in tetrahydrofliran (5 ml). The solution was stirred at room temperature for 3 hours.and then diluted with ethyl acetate, neutralised the pH with saturated aqueous sodium hydrogen carbonate solution The ethyl acetate layer was separated, dried (N&2SO4), and evaporated to dryness. The residue was purified by chromatography using a 1 Og silica isolute eluting with CH-2C25% Methanol/CH2CI2 to give N-hydroxy-N-(3-pyridin- 2-yl-lI [(4-pyridin-2-ylpiperazino)sulfonyl]rnethyl propyl)formarnide (0.039g) 80-84 0

C

NMR (d6-DMSO 373 'K):2.10 (in, 2H), 2.80 (mn, 2H), 3.25 (dd, 3.30 (in, 4H), 3.50 (dd, IH), 3.60 (in, 4H), 4.42 (vbs, LH), 6.70 (mn, 1H), 6.85 1H), 7.19 (in, 1H), 7.22 (d, 1H), 7.54 (in, IH), 7.65 (in, 1H), 8.10 (vbs, lIH), 8.15 (in, IN), 8.45 (in, IN), 9.50 (vbs, 1lH);in/z 420 Examnle 14 N-T I -r(4-If 5-chloropyridin-2-yl)ox-vlpiperidino Isulfonvl)methyll-3-uvridin-3-vloropvl hvdroxvfornainid e 00 0 N S 0-N To a solution of 1 -N-[2-(hydroxyanmiTo)-2-(3-pyridiny)butanesulfonyl]-4-O-(5chloro-2-pyridinyl)piperi dine (2.1 g, 4.18 minol) in TI-F (36 ml) added a preformed mixture of formic acid (9.0 ml) and acetic anhydride (2.25 ml). The mi xture was stirred at room temperature for 18 hrs. The reaction was neutralised using saturated aqueous NaI{C03 before extracting the solution with EtOAc The combined organics were dried over Na 2

SO

4 and evaporated in vacuo. The residue was stirred in MeOH at room temperature for hrs to remove the bis-formyl. The residue was crystallised from EtOH to afford a white solid (0.898g). m.p. 130-140 0

C.

'H NMR (DMSO-100°C) 9.50 (br s, 1H), 8.43 1H), 8.39 (dd, 1H), 8.15 1H), 8.13 (br s, 1H), 7.74 (dd, 1H), 7.60 1H), 7.27 1H), 6.83 1H), 5.12 1H), 4.32 (br s, 1H), 3.42 3H), 3.16 3H), 2.68-2.54 2H), 2.06-1.93 4H), 1.76 2H); MS 469.2 (MHW, 491.1 (MNa); EA: calculated for C 2 oH25CIN 4 OsS: C 51.22, H 5.37,CI 7.56, N 11.95, S 6.84, Found: C 50.92, H 530, Cl 7.55, N 11.90, S 6.75.

The starting material was prepared as follows i) NaH (2.88g, 66mmol, 55% dispersion in mineral oil) was stirred in dry DME (200ml), under Argon. A mixture of 2,5- dichloropyridine (8.87g, 60mmol) and 4hydroxypiperidine (6.67g, 66mmol) dissolved in dry DME (200ml) was added to the NaH suspension dropwise, over a period of 30 minutes. After complete addition the reaction is heated to 82C for 48 hrs, maintaining the Argon blanket. The reaction was slowly quenched with water before removing most of the THF. Extracted the aqueous with DCM The organic layers were dried with Na 2

SO

4 and evaporated in vacuo to afford 2-(4-piperidinyloxy) as a yellow oil (12.7g, quantitative). 'H NMR (DMSO): 8.17 1H), 7.76 (dd, 1H), 6.81 1H), 4.96 1H), 2.93 2H), 2.53 2H), 1.91 2H), 1.46 2H); .MS 213.3 225.3 (MNa) ii) To a solution 2-(4-piperidinyloxy) -5-chloropyridine(12.9 g, 0.06 mol) and Et 3

N

(25.4 ml, 0.182 mol) in dry dichloromethane (400 ml) at 0°C and under Argon, was added methanesulfonyl chloride (5.3 ml, 0.067 mol) in dry dichloromethane (100 ml), dropwise. The mixture was stirred for 20 hours at room temperature. The mixture was dilutedwith dichloromethane (250 ml), then washed with water (x3) then brine. The organic layers were dried with Na 2

SO

4 and evaporated in vacuo. The residue was triturated and washed with diethylether to give 2-(N-methanesulfonyl-4-piperidinyloxy) -5-chloropyridine (15.1 g) as a pale yellow solid.

'H NMR (DMSO): 8.20 1H), 7.81 (dd, 1H), 6.87 1H), 5.09 1H), 3.32 2H), 3.11 2H), 2.90 3H), 2.02 2H), 1.75 2H); MS 291.2 313.2 (MNa).

iii) 2-(N-methanesulfonyl-4-piperidinyloxy) -5-chloropyridine (2.0g, 6.89 Inmol) was taken into anhydrous THF (100 ml) under Argon then cooled to -78 0 C before the addition of Li(TMSA) (13.8 ml of a 1.OM solution in THF, 13.8 mmol), The mixture was stirred at -78°C for 20 minutes and a solution ofdiethylchlorophosphate (1.05 ml, 7.23 mmol) was added. The -51midxture was stirred at -781C for I hour before 3-pyridinyipropanal 12g, 8.27 inmol) was added then stirred at -78 for a fiuther I hr. The mixture was allowed to warmed to room temperature then was washed with aqueous ammonium chloride and extracted with ethyl acetate. The organic layers were washed with water, brine and dried over Na 2

SO

4 Purification of the residue on silica (eluant: gradient DCM 2% MeOHIDCM) afforded 2-fN-[IEIZ-4(3pyridyl)-but-lenyl] sulfonyl}4-piperidinyloxy) -5-chloropyridine as a yellow oil (2.09g).

1H NMR (DMSO): 8.45 (mn, 1Hi), 8.37 (in, IFI, 8.19 (in, 1K), 7.82 1K), 7.64 (mn, 1K), 7.30 (mn, 1K), 6.85 (mn, 1K), 6.88-6.27 (mn, 2H, E!Z isomers), 5.00 (mn, 1K), 3.15 (in, 2K), 2.83 (n,4 5K), 2.61 (in, IH), 1.85 (mn, 2H), 1.70 (mn, 2K); MS 408.1 (MK4), 430.2 (MNa4).

iv) To a s olution of 2-{N-[E/Z-4(3-pyridyl)-but-lenyl] sulfonyl}4-piperidinyloxy) chloropyridine(2.09 g, 5.1 mmol) in THE (20 ml) was added hydroxylainine (3.4 mld, 50 aqueous solution). The mixture was stirred for 18 hours. The solvent was evaporated. The residue was dissolved in EtOAc and washed with water The organic layer was dried on Na 2

SO

4 and evaporated in vacuo to give 2-(4-piperidinyloxy) -5-chioropyfidine 1-N-[2- (hydroxyaiino)-2-(3-pyridinyl)butanesulfonyl-4o.(-chloro-2pyridinyl)pperidine (730 mng).

'H NUR (DMSO): 8.43 1M, 8.37 (dd, 1H), 8.18 1K), 7.78 (dd, 1K), 7.61 (in, 1K), 7.36 1H), 7.29 (in, 1K), 7.85 1K), 5.70 1K), 5.08 (mn, 1K), 3.35 (mn, 3H), 3.16-3.00 (hr mn, 4K), 2.80-2.60 (hr m, 2K), 1.98 (in, 2K), 1.84 (nm, 2H), 1.69 (in, 2K); MS 441.2 463.2 (MINa4).

Using an analogous procedure to that described in Example X, a was reacted with the appropriate aldehyde to give the compounds listed below.

0 R3- [=0 o

,VN--OH

RR1 R1 R2 R3 MW MS (ES 4 Ph H 4-chlorophenyl 438 439 PhCH2CH2 H 3-chlorophenyl 466.99 468 PhCH2CH2 K 3,4-dichlorophenyl 501.43 501 PhCH2CH2 H 4-chlorophenyl 466.99 468 PhCH2CK2 H 5-chloro-2-pyridyl 467.98 468 PhCH2CH-2 K 6-chloro-4-pyrinidiyl -468.96 469 Methyl Methyl 5-chloro-2-pyridyl 391.88 392 PhCH2CH2 H 2-py-idyl 433.53 434 3 -pyridyl H 5-chloro-2-pyridyl 440.91 441 3-pyridylCH2CH2 H 5-chloro-2-pyridyl 468.96 469 2-pyridylCH2CH2 H 5-chloro-2-pyridyl 468.96 469 PhCA2OCH2 H 5-chloro-2pynidyl 483.97 .484 The following aryl-4-O-piperidines have been described previously: Piperidine, 4-(3 -chlorophenoxy)-(9C1), CAS (97840-40-9) Piperidine, 4-(4-chlorophenoxy)-(9C1), GAS (97839-99-1) Pyridine, 2-(4-piperidinyloxy)-(9C1), GAS (127806-46-6) Piperidine, 4-(3,4-dichlorophenoxy)-(9G1) was synthesised in the following alternative route:- 1) To a stirred solution 4-hydroxypiper-idine(3.5 g, 0.03 5 mol) in dry methanol (50 ml) at 0 0 C, was added dit butyl dicarbanate (9.2 ml, 0.042 rnol) in dry methanol (50 ml), dropwise.

The mixture was stirred for 20 hours at room temperature. The methanol was removedand the remaining solution was taken into Et 2 O, then washed with I M citric acid (x3) and water The combined aqueous extracts were extracted with Et2O which was dried with Na 2

SO

4 and evaporated in vacua. Purification of the residue on silica (eluant gradient, DCM MeOHIDCM) afforded N-BOC-4-hydroxypiperidine as a yellow oil 'HNM (DMS0): 4.05 3.70-3.52 (binm, 3H), 2.92 (mn, 1.66 (in, 2H), 1.40 9H), 1.33- 1. 18 (br m, 2H); MS 201.3 WMH), 219.4 (MNH 4 2) To a stirred solution N-BOC-4-hyciroxypiperidine(2.0 g, 0.01 mol), trip henyl phosphine (3.68g, 0.014 mol) and 3,4-dichlorophenol (1.96g, 0.012 inol) in dry toluene (75 ml) [with molecular sieves, at 0 0 C and under Argon] was added diethyl azodicarboxylate (2.52 ml 0.016 inol), dropwise. The mixture was stirred for l.5hrs at 0 0 C. Filtered the solution and removed the toluene before vigorously stirringin isohexane (I O0ml) and filtered the resulting suspension. The filtrate was washed with 2M aqueous NaOH dried with Na 2

SO

4 and evaporated in vacua. Purification of the residue on silica (eluant 20% Et0Acfisohexane afforded N-Boc-P iperi dine, 4-(3,4-dichloropiwnoxy)-(9C1) as a yellow solid (1 .96g). 'H NMR (DMSO): 7.52 1H), 7.31 IN), 7.01 (dd, 1H), 4.62 (mn, IH), 3.65 (mn, 2H), 3.15 (mn, 2H1), 1.88 (mn, 1.53 (in, 2H), 1.40 9H-I); MS 346.3 (M ),368.4 (MNa+).

3) 50% aquous trifluoroacetic acid (18m1) was added to a stirred solution N-Bocpiperidine, 4-(3,4-dichlorophenoxy)-(9C1) (1.96 g, 5.66 iniol). After 3.Shrs toluene is added and evaporated in vacua, this was repeated twice. The residue was then taken into EtOAc -53washed with saturated aqueous NaHCO 3 dried with Na 2

SO

4 and evaporated in vacuo to afforded'pipei-idine, 4-(3,4-dichlorophenoxy)-(9Cl) a white solid 'H NMvR (DMSO): 7.54 1K), 7.35 1K), 7.04 (dd, 1H), 4.70 (in, 1K), 3.31 (in, 2H), 3.09 2M, 2.08 (in, 2H), 1.-80 2H); MS 2.26.3 (M4H").

Piperidine, 4-(3,4-dichlorophenoxy)-(9C1) was then taken through steps ii-iv as described above.

Example 1 -Mesyl-4-(5-methoxycarbonyl-2-pyridyl)piperazine 1 -Mesylpiperazine hydrochloride (4.24g) was added to a solution of methyl 6chloronicotinate (1 .7g) and N,N-diisopropylethylamine (6.3m1) in dimethylacetainide and the mixture was heated at 120'C for 2 hours. The mixture was allowed to cool to ambient temperature and poured onto crushed ice/water (S0mI) to precipitate a tan solid. The solid was collected by filtration and dried at 80 'C for 18 hours in a vacuum oven, to give Imesyl-4-(5 -methoxycarbonyl-2-pyridyl)piperazine mp 205-207C.

NMR (d6-DMSO): 2.90 3H), 3.20 (in, 4H), 3.78 (mn, 3.80 3H), 6.92 1K), 8.00 (dd, 1K), 8.67 1K); mlz 300 Using an analogous procedure 1-inesylpiperazine hydrochloride, CAS( 16 13 57-89-7), was reacted with the appropriate chloropyridine to give the following compounds.

0 R-N N-SCH 3 R [MW mlz 6-Cl-2-pyridyl 275 276 5-CI-2-pyridyl 275 276 3 -2-pyridyl 309 310 3 -2-pyridyl 343 344 5-CN-2-pyridyl 266 267 3-CI-2-pyridyl .275 276 S-Br-2-pyridyl 320/322 -54- I -(6-chloropy-imidin-4-yl)-4- mesylpiperazine A mixture of 4, 6-dichloropyrimi dine (39. 4g), 1 -mesylpiperazine hydrochloride (55. 7g) and triethylarnine (I1 6ml) in ethanol (5O0mI) was stirred at reflux temperature for 4 hours.

The mixture was then stirred at room temperature for 12 hours. The solid, which had separated, was collected by filtration, slurry washed with ethanol (2x80m1,. 160m1) then with.

diethyl ether (1 50mI), and dried to give I -(6-chloropyfimidin-4-yl)-4- mesylpiperazine as a cream solid (71 mp 200-202*C NMvIR (d6-DMSO): 2.88 3. 18 (in, 4H), 3. 80 (in, 4H), 7.04 111), 8.3 8 (mn, IN); m/z 277.3 Using an analogous procedure I -mesylpiperazine hydrochloride, CAS( 161357-89-7), was reacted with the appropriate chioropyrimi dine or chioropyridazine to give the following compounds.

0 N 11-C R MW inlZ 2-CI-pyrimidin-4-yl 276 277 6-CI-pyridazin-3-yl 276 277 pyrimidin-4-yl 242 243 6-ehoxy-pryrimidin-4-yl Example 16 o 0 C1 N S.I N Acetic anhydride (19m1) was added directly to formic acid (76m1). The solution was stirred at room temperature for 30 minutes. A solution of 1-(6-chloropyrimi din-4-yl)-4-{[2- (hydroxyamino)-4-phenylbutyl]sulphony piperazine (1 7.2g) in tetrahydrofuran (S Smi) was added in portions, to the above solution at 27 °C over 25 minutes. The solution was stirred at room temperature for 1 hour. The solution was evaporated (water-bath temperature 30 0

C)

and the residual gum was dissolved in ethyl acetate (500ml). This solution was treated with saturated aqueous sodium hydrogen carbonate solution (200ml) and the mixture (pH8) was stirred at room temperature for 16 hours. The ethyl acetate layer was separated, washed with saturated brine (100ml), dried (Na 2

SO

4 and evaporated to dryness. The residual foam was dissolved in ethanol, a solid separated and the'mixture was stirred for 2 days. The solid was collected by filtration, slurry washed with diethyl ether (100ml), and dried to give (6-chloropyrimidin-4-yl)piperazino]sulphonyl}methyl)-3-phenylpropyl]-N-hydroxyformamide as a colourless solid (12.8g). mp 155-157C.

Found C, 50.29, H, 5.29, Cl, 7.82, N, 15.31, and S, 6.82 C 19

H

2 4

CIN

5 0 4 S requires C, 50.27, H, 5.33, Cl, 7.81, N, 15.43, and S, 7.06 NMR (d6-DMSO 373°K): 1.93 1H), 2.03 1H), 2.57 1H), 2.65 1H), 3.20 (dd, 1H), 3.26 4H), 3.48 (dd, 1H), 3.74 4H), 4.3 (v br, 1H), 6.90 1H), 7.19 3H), 7.27 2H), 8.1 (br, 1H), 8.38 1H), 9.5 1H); m/z 454.2 O0 S 0 CI N N N Acetic anhydride (31.5ml) was added directly to formic acid (126ml). The solution was stirred at room temperature for 30 minutes.

A solution of [3-benzyloxy-2-(hydroxyamino)propyl]sulphonyl}-4-(6-chloropyrimidin-4yl)piperazine (29.5g) in tetrahydrofuran (150ml) and formic acid (25ml), was added in portions to the above solution at 25 °C over 25 minutes. The solution was stirred at room temperature for 1 hour. The solution was evaporated (water-bath temperature 300C) and the residual gum was dissolved in ethyl acetate (500ml). This solution was treated with saturated aqueous sodium hydrogen carbonate solution (2x250ml) and the mixture (pH8) was stirred at room temperature for 16 hours. The ethyl acetate layer was separated, washed with saturated brine (100ml), dried (Na 2

SO

4 and evaporated to dryness. The residual foam was dissolved in methanol (70ml) and the solution was stirred for 16 hours. The solution was evaporated to dryness (water-bath temperature 30 The residual foam was stirred in ethanol (250ml), -56solid separated and the mixture was stirred for 1 8 hours. The solid was collected by filtration, slurry was .hed with diethyl ether (I G0mi), and dried to give N-[2-(benzyloxy)-I1-([ chloropyrimidin-4-yl)piperazino]sulphony }methyl)ethyl]-N-hydroxyformamide (25 mp I 18-120 0

C

Found C, 48.35, H, 5.09, Cl, 7.26, N, 14.73, and S, 6.78 CigH 24 ClNsO5S requires C, 48.56, H, 5.15, Cl, 7.54, N, 14.90, and S, 6.82 NMR (d6-DMSO 373 3.23 (dd, 1H), 3.30 4H), 3.46 (dd, lH), 3.57 (dd, IH), 3.67 (dd, I 3.72 4H), 4.5 0 2H), 4.5 0 (in, I 7.3 5 (rn, 8.15 (br, IH), 8.38 IH), 9.48 (br, IH); ma/z 470.2 0 0

N

Acetic anhydride (0.8m1) was added directly to formic acid (3.2m1). The solution was stirred at room temperature for 30 minutes.

A solution of I -(5-chloro-2-pyridyl)-4- {[2-(hydroxyamino)-4phenylbutyljsulphonyl)piperazine (0.72g) in tetrahydrofliran was added to the above solution at room temperature. The solution was stirred at room temperature for 2 days. The solution was evaporated (water-bath temperature The residue was dissolved in 5% methanol in dichloromethane. Silica (5g Merck 9385) was added to the solution, the mixture was stirred for 21 hours, and evaporated to dryness. The material (pre-adsorbed on the silica) was purified by chromatography on silica (Bond Elut I Og), using 0-3% methanol in dichioromethane as eluent, to give I chloro-2-pyridyl)piperazino] sulphonyl }methyl)-3 -phenylpropyl]-N-hydroxyformamide as an orange foam (0.17g).

NUR (d6-DMSO 373K): 1.92 (in, 1H), 2.04 (in, 1K), 2.55 1H), 2.64 (in, 1m), 3.20 (dd, 1H), 3.27 (in, 4H), 3.47 (dd, 1K), 3.58 (in, 4H), 4.35 (v br, 6.88 (dd, 1H), 7.17 (in, 7.27 (in, 2H), 7.57 (dd, IH), 8. 10 lH), 8. 10 (br, 1K), 9.5 lH); in/z 453.3 Example 17 H

H

0 a N N N N-S O

O

o Nb 0 To formic acid (31.5ml) at 0°C was added acetic anhydride (7.9ml). After 20 minutes this was added to the hydroxylamine (6.10g) dissolved in THF (80ml) and formic acid and the resulting solution stirred overnight at room temperature. The solvent was removed under reduced pressure and the residue dissolved in DCM (500ml), washed with saturated sodium bicarbonate solution (2x500ml), dried and evaporated to dryness. To the residue dissolved in DCM (10ml) was added diethyl ether (100ml) to give the product as a white solid (5.60g) which was collected by filtration. Mpt 168-170 0 C. NMR DMSOd 6 d 10.2 (br s, 9.8 (br s, 8.7 (br s, 8.6 (br s, 8.5 1H); 8.3 1H); 8.1 (d, 7.9-7.8 1H); 7.6 (dd, 1H); 7.4 (dd, 1H); 6.9 IH); 5.8 5.5 1H)*; 4.1-3.6 2H); 3.6 4H); 3.2 4H). Anal. Calcd for C 1 7

H

20 CINsO4S: C, 48.0; H, 4.7; Cl, 8.3; N, 16.5; S, 7.5. Found: C, 47.9; H, 4.7; Cl, 8.4; N, 16.3; S, 7.5. MS for

C

1 7

H

2 0 ClN 5 0 4 S calcd 426, found 426.

rotameric signals Step A OH -O S-NOH 0 (1) The oxime (31.05g) [Tetrahedron Letters 1994, 35, 1011 was dissolved in DCM S(500ml) and 3-pyridinecarboxaldehyde (12.09g) was added followed by anhydrous -58magnesium sulfate (13.6g). After 2 days stirring at room temperature more magnesium sulfate (13.6g) was added and stirring was continued for a further 3 days. The mixture was then filtered, the solvent evaporated and the residue triturated with diethyl ether to give the product (36.34g) as a white solid. Mpt 174-175 0 C. NMR CDC13 d 9.0 1H); 8.9 1H); 8.7 1H); 7.7 IH); 7.4 (dd, 1H); 5.6 1H); 5.3 1H); 4.9 (dd, 1H); 4.6 (dd, 1H); 4.4 (ddd 1H); 4.2 (dd, 1H); 3.7 (dd, 1H); 1.5 3H); 1.4 3H); 1.4(s, 3H); 1.3 3H).

Step B r OH SN ONNC. N 0 L\ ,N N Cl (2) The methyl sulfonamide (14.30g) was dissolved in THF (500ml) and cooled to when lithium hexamethyldisylazide (78ml, 1.OM in THF) was added. After 30 minutes the Ssolution was cooled to -78 0 C, and the nitrone (18.00g) dissolved in THF (350ml) was added, keeping the temperature below -65 0 C. The resulting solution was stirred for 3 hours at -78 C when it was quenched by the addition of brine (500ml) and the aqueous layer extracted with ethyl acetate (3x500ml). The combined organic layers were dried and evaporated to give a yellow solid which was triturated with ethyl acetate isohexane and then purified by flash column chromatography eluting with dichloromethane methanol (97:3) to give 1 (16.40g) as a white solid. Mpt 209-211°C (dec). NMR CDCI1 d 8.6 1H); 8.4 1H); 8.1 1H); 7.8 1H); 7.5 (br s, 1H); 7.4 (dd, 1H); 7.3 (dd, 1H); 6.6 1H); 4.9 1H); 4.8 1H); 4.7-4.6 2H); 4.2-4.1 3H); 3.8 (dd, 1H); 3.6 (dd, 1H); 3.5-3.4 5H); 3.3- 3.2 4H); 1.4 3H); 1.3 3H); 1.3 3H); 1.3 3H).

-59-a Step C

OH

0' NN 0 0 I K ,N IN N 014 (3) To a solution of hydroxylamine 2 (14.90g,) in ethanol (300rn1) was added water (220m1) followed by O-benzylihydroxylamine hydrochloride (13.91 g) and sodium bicarbonate (6.95 Heating gave a solution which was stirred overnight at 80"C. The ethanol was removed under reduced pressure and the residue separated between water (500m1) and ethyl acetate (500m1). The aqueous layer was washed with ethyl acetate (2x500m1), and the combined organic layers were dried and evaporated to give a residue which was triturated with dichloromethane (lO0mi) to give 3 (6.10g) as a white solid. The mother liquor was purified by flash column chromatography eluting with ethyl acetate followed by dichloromethane methanol (96:4) to give further 3 (0.85g). Mpt 170-1 73 0 C. NMvR D MS Od 6 d 8.6 I S.5 I1H); 8. 1 7.89 I1H); 7.6 (dd, I 7.6 1KH); 7.3 (dd, 1K); 6.9 IH); 6.1 (br s,1KH); 4.3 (br s, 3.7-3.4 (in, 6H); 3.2-3.1 (in, 4K).

Examnle 18 The following compounds were made using the method outlined in Example 7 ~B-A A-Y-QR 2

R

Low HI h jMB j A Y IQ RI Z I 403 14-PhCH2 IPi eridinvI S02 ICH2 Ph JRH 1 357 14-HCOO lPiperidinvi IS02 CH h RH PhNCO jPiperidinvi S02 JCH jhIRH 128 j131 1412 It-ButvlNCO IPiperidirIv IS02 ICH j RH MP MP M+H *Low-Hi h B A Y Q RI

Z

122 124 446 PhCH2NCO Pi eridinvl 502 CR2 Ph

RH

129 131 423 c-PentvlNCO'r Pi -n'dinvi S02 CH2 Ph

RH

390 Ph PIP S02 CR2 Ph

R

420 4-MeO-Ph PIP S0CRPhH 435 4-N02-Ph PTP S0CRPhR P0P SO2CCphRI- 396 CI-h~y PIP 02 C2 Ph

R

46 3-h-Ph PIP S02 CH2 Ph

R

4 4-C3 -h PIP S02 C2 Ph

RH

47 4-C3-PhC PIP 02 C2 Ph

RH

44 2--Nhh PIP S02 C2 Ph

RH

356 n-o viP PIP S02 C2 Ph

RH

48 4-i eo-C- PIP S02 C2 Ph

R

4--utl-PhCH2 PIP S0l CR2,h"R 60 4 52 4-CI-PhC P edn v 50 R hR 391 4-Pidy PP S02 IC2 Ph

R

44 4-4-2 -ph Pip S2 ICH2 Ph

R

42 4Ph-O-ph PIP 502 CH2 Ph

R

404 4-hhv 3-OoPEP S02 CH2 Ph

RH

49 5-O2-Pvnv PIP 02 C2 Ph

RH

301 2-PridiNC Pip Srdnv 02 C Ph-i-IPhR 5396 5CI-vc INC Piiv 502 C Ph-i-IPhR 54 4-CF-PC PIP eSdv S2 C2 JAdPh

RH

500 4lPhNCO Pipeiiv S02 C2 34dIPh

RH

356 n hyodrIP S02genP 448 4-P criperH IPz02inyPlP 46 RI-Itl-hC2 PI Sevrs hydroPamate The starting material was prepared as follows: The addition ofhydroxylamine to I-trans-P-styrenesulphonyl-piperidine-4-(Nphenylcarboxamide) and the subsequent formylation of the product was carried out as described in Example 7.

Dimethylformamide (2 drops) was added to a suspension of 1-trans-pstyrenesulphonyl-piperidine-4-carboxylic acid (0.75g) and oxalyl chloride (0.23 mL) in dichloromethane (10 mL) and was stirred for 2 hours. The reaction mixture was evaporated to dryness, re-dissolved in dichloromethane (10 mL) and evaporated to dryness again. The residue obtained was dissolved in dichloromethane (4 mL) and a mixture of aniline (0.23 mL) and triethylamine (0.35 mL) was added dropwise. The mixture was stirred for 20 hours and was washed with dilute 2M hydrochloric acid, water, aqueous saturated sodium bicarbonate solution and water and dried. Removal of the solvent gave 1-trans-P-styrenesulphonylpiperidine-4-(N-phenylcarboxamide), 0.89g Using the method described above there were prepared the following 1 -trans-P-styrenesulphonyl-piperidine-4-carboxamides -62- A solution of ethyl piperidine-4-carboxylate (3.99g) in a mixture of THF (30 mL) and methanol (6 mL) was treated with aqueous sodium hydroxide solution (20 mL of2M NaOH) and the mixture stirred for 3 hours, evaporated to small volume and acidified to pH 5 with.

dilute 2M hydrochloric acid. The mixture obtained was extracted with ethyl acetate (2x25 mL), the ethyl acetate extracts were washed with water, dried and evaporated to dryness to give 1-trans-P-styrenesulphonyl-piperidine-4-carboxylic acid, 2.64 g.

A solution of ethyl piperidine-4-carboxylate (3.0 mL) and triethylamine (2.7 mL) in dichloromethane (10 mL) was added dropwise to a cooled (ice bath) solution oftrans-3styrenesulphonyl chloride 3 .95g) in dichloromethane (10 mL). The reaction mixture was allowed to warm to ambient temperature and stirring was continued for 20 hours. The reaction mixture was evaporated to dryness the residue was diluted with water and extracted with ethyl acetate (2x25 mL). The combined ethyl acetate extracts were washed with brine and dried (MgS04) to give ethyl -(1-trans--styrenesulphonyl)-piperidine-4-carboxylate 5.76g, M+H 324.

An alternative procedure for the preparation of 1-trans-3- 3, 4 dichlorostyrenesulphonyl-piperidine-4-carboxylic acid may be used: To a solution of 1-trans-P- 3 ,4dichlorostyrenesulphonylchloride (2.7g) and isonipecotic acid (l,41g) in acetonitrile (15ml) was added 2M sodium hydroxide (11ml) and stirred at ambient temperature for 1 hour. The reaction mixture was acidified to pH 3 with 2M hydrochloric acid and extracted with ethyl acetate (2x15ml), the ethyl acetate extracts were dried (Na 2

SO

4 fitered and evaporated to give 1-trans-P-3,4dichlorostyrenesulphonylpiperidine-4-carboxylate (2.67g), m/z 364 Example 19 The following compounds were prepared R1 PIP piperazinyl Z reverse hydroxamate group R2 hydrogen M+H B A Y Q RI Z 4-F-Ph PIP S02 CH2 i-Propyl RH 360 4-F-Ph PIP S02 CH2 Ethyl RH 386 4-F-Ph PIP S02 CH2 spiro-c-pentyl RH 450.8 4-F-Ph PIP S02 CH2 14-NMe2-Ph RLH 442 4-F-Ph PIP S02 CH2_ 4-Cl-Ph RH 388 4-F-Ph PIP S02 CR.2 -'tert-Butyl RH 442 .4-F-Ph PIP S02 CR2 12-Cl-Ph RH 484 4-F-Ph PIP S02 CR2 4-Ph-Ph RH 468 4-F-Ph PIP S02 CHR2 2,4-di-OMe-Ph RH 452.9 4-F-Ph PIP S02 CR2 3-N02-Ph RH 475.9 4-F-Ph PIP S02 CR2 4-CF3-Ph RH 475.9 4-F-Ph PIP S02 CR2 2-CF3-Ph RHU 374 4-F-Ph PIP S02 CR2 Propyl 458 4-F-Ph PIP S02 CH2 I -Naphthyl 387.9 4-F-Ph PIP S02 1CR2 3-Furyl 450.9 4-F-Ph PEP S02 CH2 jCH2CH2SCH3 TRH 388 4-F-Ph PIP S02 CR2 liso-Butyl RH 491.8 4-F-Ph PIP S02 CR2 14-Br-2-Thiophenyl RH 485.8 4-F-Ph PIP S02 CR2 13-Br-Ph RH 458 4-F-Ph PIP S02 CR2 2-Naphthyl RH 496 4-F-Ph PIP S02 CR2 2-Fluorenyl RH 466 4-F-Ph PIP S02 CR2 4-CO2Me-Ph RH 414 4-F-Ph PIP S02 CH2 Cyclohexyl RH 402 4-F-Ph -PIP S02 CH2 2-neopentyl RH 533.9 4-F-Ph PIP S02 CH2 3-(4-Cf-PhO)-Ph RH 452 4-F-Ph PIP S02 CRTH2 PhCH2OCH2 RH 507.9 4-F-Ph PIP S02 I CH2. 2-(5-4'-CI-Ph)Furyl RH 450 4-F-Ph PIP S02 CR2 CH2CH(CH3)Ph RH 451.9 4-F-Ph* PIP S02 CR2 4-Piperonyl RH PIP S02 CR2. 3-(OCH2Ph)Ph RH PIP S02 CR2 4-(OCH2Ph)Ph RH +4-F-Ph PP S02 C23:-F3-Ph

RH

497.9 4-F-Ph PIP IS02 .ICR2 JC6 F5 RHd -64- Example We-provide NNM data for the following compounds: F

NN-S

o N 0-N (DMSO) 9.6 (1K1 8.5 (1KL 8.4 and 7.9 (1K 7.7 (1K in), 7.2 (211 mn), 7.1 (21K mn), 7.0 (2H, in), 4.7 and 4.2 (1K broad in), 3.4 (111 in), 3.3 (5HK in), 3.1 (4K1 in), 2.7 (21L, in), 0 N N-S 0 (DMSO) 9,8 and 9.5 (1K, broad 8.3 and 8.0 (1K, 8.1 (1K 7.6 (111, dd), 7.2 (511, mn), 6.9 (111,L 4.7 and 4. 1 (1 H broad in), 3.6 (4KL in), 3.4 (1KH, 3.3 (1 H, mn), 3.2 (4K, mn), 2.6 (2H, in), 1.6(41-L in).

0 clI N N N 1 (DMSO) 9.6 (1KL broad 8.4 (1K in), 8.3 and 7.9 (11, 8.1 (IH, 7.6 (2-L in), 7.2 (1K 7.1 (1K, in), 6.9(IK 4.7 and 4.1 (1K broad mn), 3.6 (4H, nm), 3.4 (1K mn), 3.3 (I1K 3.2 (4K in), 2.7 (2H, in), 2.0 (2K, in).

-0 Nt N N-S N 0 0 (DMSO) 9.7 (Ii, broad 8.5 (1,L in), 8.4 (1K mn), 8.1 and 7.9 (1KL 7.6 (1K, in), 7.2 (2H, 7.0 (1H, in), 4.6 and 4.1 (1K broad in), 3.7 (4K 3.4 (1Ki mn), 3.3 (5H, mn), 2.7 (2K, 2.0 (2H, in).

Ni 0 (DMSO) 9.9 (1H, 8.4 (2H, mn), 8.2 (1K, 7.65 (2H, in), 7.3 7.0 4.2 (2H, mn), 3.6 (41-L br mn), 3.4-3.2 (6K br in), 2.0 (21K, br mn).

N

(DMSO) 10.0 (1K, 8.5 (2H, 8.2 (1K, br 7.8 (1K br), 7.6 (1K, in), 7.4 (1K 6.9 (1KH, mn), 3.6 (41H, br in), 3.2 (6H, br in).

0 F 0 F N N-S 0 0 (1IH, 8.5 (2K 8.4 and 8. 0 (1K 7.9 (1KH, mn), 7.7 (1K inL 7.3 (1K In 7.1 (1K 3.7 (4H, br mn), 3.45 (2K mn), 3.3 (4H, br mn), 2.75 (3K, mn), 2.1 (2K in).

F0 -66- (DMSO) 10. 0 (lIi br 8. 6 (2-L mn), 8.2 (1 H, 7.2 (1H Rin), 6.9 (4H, in), 4.9 and 4.2' (111, br), -3.4 (61-L mn), 3.0 (6HK 1.9 (4K1 in).

N

N

I-

(DMSO) 9.8 (111, br), 8.7 (2H, nm), 8.3 and 7.9 (11, 8.1 (2H, 7.6 (111, mn), 7.3 (111, m), 6.9 (11H, mn), 4. 1 (11H, br mn), 3.6 (4K, 3.2 (6H, mn), 2.8 (2H, mn), 1. 8 (411, in).

0

'I

N 0 100

(DCI

3 8.5 8. 5 and 8. 0 (111, 7.(111, in), 7. 4 (16H, m 7 (11, 6. 11 n,48ad42(1,b n,3.6 (411 in) 3.2 (61 in), 2.8 (211 1.8 (411, n).

Exampnle 21 The folowing compounds were prepared PIP piperazinyl Z =reverse hydroxamate group R2 hydrogen Mp IMp M+H Low High B A Q_ I RI Z 1 467 4-Cl-Ph PIP 02 CH2 3-PyridylCH(CH3)CH2 RH 16() 456 4-F-Ph PIP S02 CH2 c hexvlC(Me)CH2 RH 125 128 440 4-F-Ph PIP S02 CH2 PhCH2SC142 RH 130 131 1460 4-F-Ph Piperidirivl S02 CR2 2-IndanCH2L RH 64 65 448 4-F-Ph Piperidinyl S02 CH2 (R)-2-PhCH(CH3)CH2 RH 63 64 448 4-F-Ph Piperidinvl S02 CH2 I(S)-2-PhCI-I(CH3)CH2 RH.

132,137 484 4-F-Ph PIP S02 CR.2 2-Cl-PhCfI(CH3)CH2 RH 484 4-F-Ph PIP S02 CH2 4-CI-PhCH(CH3)CH2 RH 484 4-F-Ph PIP S02 CR2 3-C1-PhCI-I(CH3)CH2 RH 469 5-Cl-2-Pyridyl PIP S02_CR2 2-PvrazineCH(CH.3)CH.2 RH 516 4-F-Ph PIP S02,CH2 4-CI-Ph-S-CH(CH3)CH2 _RH 466 3-Cl-Ph PIP S02 CR2 (S)-2-PhCHq(CH3)CH RH 467 5-CI-2-Pyridyl PIP S02_CH.2 (S)-2-PhCH(CR3)CH2 RH 151 450 4-F-Ph Piperidinyl S02_CH2- 2-PyrazineCH(CH3)CH2 RH 61 454 4-F-Ph Piperidinyl S02 CH2 2-ThiophenylCH(CH3)CH2 RH 82 83 449 4-F-Ph PiperidinyI S02, CR2 14-PyridyCH(CH3)CH-2 RH 66 407 4-F-Ph PIP S02 ICR-Ph RH 91 100 484 4-F-Ph PIP S021CH2 PhiCH2SOCI2 RH 142 145 484 4-F-Ph PIP S02 CR2 PhCH2SOCH2 RH 455 5-CI-2-Pyidyl PIP S02 CH2 2-PvrimidinylCH2CH2 RH 460 5-cyano-2-pyridyl PIP S02 CR2 2-PymnidinylCR2CH2CH2 RH 44-4 5-cyano-2-pyridyl PEP S02 CR2 PhCH.2CR2 RH S 464 5-cyano-2-pyridyl PIP S02.CR2 2-ThiophenylCH2CH2CH.2 RH 445 5-cyano-2-pyridyl PIP S02 CR2 3-PyridylCH2CH2 RH 459 5-cyano-2-pyfidyl PIP S02 CR2 2-PyridylCHEMHCH2CH2 RH 460 5-cyano-2-pyridyl PIP S02 CR2 PhCH.20CH2 RH 445 5-cyano-2-pyridyl PIP S02 CR2 2-PyridylCH2CH2 RH 417 5-cyano-2-pyridyl PIP S02 CR2 3-Pyridyl RH 498/ 5-Br-2-Pyridyl PIP S02 CR2 2-PyridylCH2CH2 RH ___500 498/ 5-Br-2-Pyiidyl PIP S02 CR.2 3-PyridylCH2CR2 RH 497/ 5-Br-2-Pyridyl PIP S02 CH.2 PhCH2CH2 RH 1 499 513/ 5-Br-2-Pyridyl PIP S02 CR.2 lPhCH2OCH2 RH 515 1470/ 5-Br-2-Pyridyl PIP S02 CR2 13-Pyridyl

RI-

472 11 -68- Mp Mp M+H Low High B A -Y Q RI Z 517/ 5-Br-2-Pyridyl PIP S02 CR 2-ThiophenylCH2CH2CH2 RH 519 513/ 5-Br-2-Pyridyl PIP S02 CH2 2-PyrimidinylCH2CH2CH2 RH 515 512/ 5-Br-2-Pyridyl PIP S02 CH2 2-PyfidI1CH2CH2CH2 RH 1 514 436 2-Pyrazinyl PIP S02 CR2 2-PyrimidinvICH2CH2CH2 RH 439 2-Pyridyl PIP S02 CR2 2-Thiopheny1CH2CH2CH2 RH 440 12-Pyrazinyl PIP S02,CR2 2-Thiophe v1CH2CH2CH2 RI! 488.1 5-CI-2-Pyridyl 4-0-Piperid- S02 CR2 2-Thiopheny1CH2CH2CR2 RH 103 104 484.1 5-CI-2-Pyridyl 4-0-Piperid- S02 CR2 2-Thiopheny1CH2CH2CH2 R1H 483.3 5-CI-2-Pyridyl 4-0-Piperid- S02 CH2 2-Pyridy]CH2CH2CH2 RH iny! 508.1 5-CI-2-Pyridyl 4-0-Piperid- S02 CR2 3,4-di-Cl-Pli

RH

I inyl 504/ 15-C1-2-Pyridyl PIP S02 CH2 3-Pyridyl-5-bromo RH- 506 123 125 466.3 16-MeO-4-Pyrirnidinyl PIP S2C2 PhCH2OCH2 RH 99 101 451.3 16-MeO-4-Pvrimidinyl PIP S02 CH2 2-PvridvlCH2CH-2 RH 99 451.4 16-MeO-4-Pyrirnidinyl PIP S02 jCEZ 3-PyridvICH2CH2 RH 156 158 470.3 16-MeO-4-Pyrimidinyl PIP S02 [CR2 2-Thiophenyl1CH2CH2CH.2 RH 122 -24 466.3 16-MeO-4-Pyrimidinyl PIP S02 CH2 2-PyrimidinylCH2CH-2CH2 R.H E465.3j6-MeO-4-Pyrimidinyl PIP S02 [CR2 2-Pyridy1CH2CH2C2 JRHI All compounds were prepared as in Example I except those where ring A is piperidinyl which were prepared as in Example 14.

Example 22 We provide NNM data for the following compounds listedi in Example 2 1: CI- -N N-S N 0 N 0 N' C I -N IE~~ 0 0-N M452587 new compound NMIR DMSO 9.9,9.6 (1KH broad s 8. 6 2H 8.3 and 7.9 1H 8.1 1H,dd); 7.3 (1H,nm 6.9 (1H.,d 4.7 and 4.2 (1H broad,in) 3.6 4K im); 3.4-3.2 2.8 (2H, m 1 (21Hm (DMSO) 9.9 and 9.6 (1KL broad-s), 8.7 (2H, d), 8.3 and 8.0 (1KL 8.2 (1K, 7.8 (111, dd), 7.3 (1H, in), 6.9 (1K, 5.1 (1H, broad in), 4.7 and 4.1 (1K, broad in), 3.4 (3K, mn), 3.1 (3H, mn), 2.9 (2H, in), 2.0 (2H, in), 1.7 (6H, mn).

-69- Example 23 Preparation of: H O 0 N

N

jINI

H

0 N

F

N N 2 To formic acid (4.8ml) at 0°C was added acetic anhydride (1.2ml). After 20 minutes this was added to the hydroxylamine 2 (0.68g) dissolved in THF (11ml) and formic acid ,and the resulting solution stirred overnight at room temperature. The solvent was removed under reduced pressure and the residue dissolved in DCM (100ml), washed with saturated sodium bicarbonate solution (2x100ml), dried (MgSO 4 and evaporated to dryness. The residue was purified by flash column chromatography eluting with dichloromethane methanol (96:4) to give the product (0.41g) as a gum. NMR CDC1 3 6 9.7 (br s, 9.2 (br s, 8.4 8.0 7.5-7.2 5H); 7.0-6.8 4H); 5.7 5.4 (m, 3.9-3.4 5H); 3.3 3.2-2.9 4H); 2.8 MS for C 20 H22FN 3 0 3 calcd 372, found 372.

rotameric signals Step A F N NH F/N N To 1-(4-fluorophenyl)piperazine (1.00g) dissolved in DCM (10ml) was added cinnamoyl chloride (0.85g) in DCM (10ml) followed by triethylamine (1.55ml). The solution was stirred at room temperature overnight.. It was then separated between DCM (150ml) and water (100ml), the organic layer was then washed with water (100ml), dried (MgSO 4 and evaporated to dryness to give a cream solid which was triturated with diethyl ether to give 1 (1.20g) as a white solid. NMR CDC1 3 6 7.7 1H); 7.5 2H); 7.4 (m, 3H); 7.0-6.9 5H); 4.0-3.8 4H); 3.1 4H). MS for C 19

H

1 9

FN

2 0 calcd 311, found 311.

Step B

H

0 0 0 N'H FNN N 1 2 To the amide (2.00g) dissolved in THF (40ml) was added hydroxylamine (1ml, aqueous solution). The solution was stirred at room temperature for 48 hours. The solvent was then evaporated under reduced pressure, toluene was added (50ml) and this was also evaporated under reduced pressure. The residue was triturated with dichloromethane methanol (98:2) and the mother liquor purified by flash column chromatography eluting with dichloromethane methanol (98:2) to give 2 (0.70g) as a gum. NMR CDC1 3 5 7.5-7.2 (m, 7.0-6.9 2H); 6.9-6.8 2H); 4.5 (dd, 1H); 3.8-3.7 2H); 3.6-3.5 2H); 3.1- 2.8 5H); 2.7 (dd, 1H). MS for C 1 9

H

22

FN

3 0 2 calcd 344, found 344.

It will be understood that the term "comprises" or its granmatical variants as used herein. is equivalent to the term "includes" and is not to be taken as excluding the presence of other elements or features.

Claims (12)

1. A compound of formula I B A 6 V ^Z \C2 Z R3n R1 R2 wherein ring B is a phenyl, pyridyl or pyrimidyl; each R3 is independently selected from hydrogen, halogen, N02, COOR wherein R is hydrogen or C 1 6 alkyl, CN, CF 3 C1-6 alkyl, -S-Ci.e alkyl, -SO-C-6 alkyl, -S02-C1-6 alkyl, C1-6 alkoxy and up to Cio aryloxy, n is 1, 2 or 3. P is Ring A is piperazinyl. both X, and X 2 are N; Y is -S02-; Z is -N(OH)CHO; Q is-CH 2 R1 is phenyl, 4-trifluoromethylphenyl, phenethyl, phenpropyl, isobutyl, cyclopentyl, benzyloxymethyl, 3,4-dichlorophenyl, pyridyl, pyridylethyl, thiophenylpropyl, bromothiophenyl, pyrimidinylethyl, pyrimidinylpropyl, pyridylethyl, pyridylpropyl, or together with R2 is spirocyclohexane or spiro-4-pyran; R2 is hydrogen; and wherein any alkyl groups outlined above may be straight chain or branched or a pharmaceutically acceptable salt or in vivo hydrolysable precursor thereof.

2. A compound as claimed in claim 1 wherein: R3 is hydrogen, halogen, NO2, CF 3 1-4 alkyl, and C14 alkoxy, n is 1 or 2; or a pharmaceutically-acceptable salt or in vivo hydrolysable precursor thereof.

3. A compound as claimed in claim 1 wherein: R3 is hydrogen, halogen, NO2, CF 3 methyl, ethyl, methoxy, or ethoxy; R1 is phenyl, 4-trifluoromethylphenyl, phenethyl, phenpropyl, isobutyl, cyclopentyl, benzyloxymethyl, 3,4-dichlorophenyl, 2-pyridyl, 3-pyridyl, 2- pyridylethyl, 3-pyridylethyl, thiophenylpropyl, bromothiopheneyl, 2-pyrimidinylethyl, 004390879 -72- 2-pyrimidinylpropyl, pyridylpropyl or together with R2 is spirocyclohexane or spiro-

4-pyran. 4. A compound of the formula 1 as claimed in claim 1 wherein ring B substituted by R3n is a phenyl, 3-methylphenyl, 4-fluorophenyl, 3-chlorophenyl, 4- chlorophenyl, or 3,4-dichlorophenyl ring or is 5-chloro-2-pyridyl; or a pharmaceutically-acceptable salt or in vivo hydrolysable precursor thereof.

A compound of formula I as claimed in claim 1 wherein ring B is a phenyl, 3-methylphenyl, 4-fluorophenyl, 3-chlorophenyl, 4-chlorophenyl, or 3,4- dichlorophenyl ring or 5-chloro-2-pyridyl; and R1 is phenyl, or a pharmaceutically- acceptable salt or in vivo hydrolysable precursor thereof.

6. A pharmaceutical composition which comprises a compound of the formula as claimed in claim 1 or a pharmaceutically acceptable salt or an in vivo hydrolysable ester and a pharmaceutically acceptable carrier.

7. A method of treating a metalloproteinase mediated disease condition which comprises administering to a warm-blooded animal a therapeutically effective amount of a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable precursor thereof.

8. A process for preparing a compound of the formula according to claim 1 or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof which process comprises a) reacting a compound of the formula (II) or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof with a compound of the formula (III) B A R3n R1 R2 004390879 -73- wherein X 1 represents X or a precursor of X (whether by modification or displacement) or an activated form of X suitable for reaction with Y'; Y' represents Y, a precursor of Y, or an activated form of Y suitable for reaction with X1', Z' represents a protected form of Z, a precursor of Z (whether by modification or displacement of or an activated form of Z; R 6 B A R1 2 1- SO--CH 2 IX R3n R 6 B A R, X P--X2 X1- SO2 or b) reacting a compound of the formula (IV) or a pharmaceutically acceptable salt or in viv hydrolysable ester thereof with a compound of the formula l- V IR B A z* XI P--X2 Xi- SO 2 XI R3n or b) reacting a compound of the formula (IV) or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof with a compound of the formula I 004390879 -74- wherein B 1 represents a suitable ring function or substituent group for reaction with P 1 Z 1 is as hereinbefore defined; and P 1 represents a suitably activated form of the linker P for reaction with B 1 or where X2 is N then P' may be present on ring A rather than ring B or, as required, the linker P may be formed by appropriate reaction of precursor groups P" and provided on rings B 1 and A respectively, or vice versa.

9. The use of a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable precursor thereof in the preparation of a medicament for use in a disease condition mediated by one or more metalloproteinase enzymes.

The use of a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable precursor thereof in the preparation of a medicament for use in the treatment of arthritis.

11. The use of a compound of the formula or a pharmaceutically acceptable salt or in vivo hydrolysable precursor thereof in the preparation of a medicament for use in the treatment of atherosclerosis.

12. A compound according to claim 1 substantially as hereinbefore described with reference to any one of the examples. AstraZeneca AB By their Registered Patent Attorneys Freehills Carter Smith Beadle 12 November 2003