ZA200606429B - Contrast agents for myocardial perfusion imaging - Google Patents

Description

CONTRAST AGENTS FOR MYOCARDIAL PERFUSION IMAGING

The present disclosure relates to novel compounds comprising imaging moieties, and their use for diagnosing certain disorders in a patient.

Mitochondria are membrane-enclosed organelles distributed through the cytosol of most eukaryotic cells. Mitochondria are especially concentrated in myocardium tissue.

Complex 1 (“MC-1") is a membrane-bound protein complex of 46 dissimilar subunits. This enzyme complex is one of three energy-transducing complexes that constitute the respiratory chain in mammalian mitochondria. This NADH- ubiquinone oxidoreductase is the point of entry for the majority of electrons that traverse the respiratory chain, eventually resulting in the reduction of oxygen to water (Q. Rev. Biophys. 1992, 25, 253-324).

Known inhibitors of MC-1 include deguelin, piericidin A, ubicidin-3, rolliniastatin-1, rolliniastatin-2 (bullatacin), capsaicin, pyridaben, fenpyroximate, amytal, MPP+, quinolines, and quinolones (BBA 1998, 1364, 222-235).

The present disclosure is based, in part, on the recognition that interrupting the normal function of mitochondria could advantageously concentrate certain compounds in the mitochondria, and hence in the mitochondria-rich myocardium tissue. If these compounds were labeled with an imaging moiety, such a build up could be detected, thereby providing valuable diagnostic markers for myocardial perfusion imaging. For purposes of this specification, a compound is referred to as «“|abeled” when an imaging moiety is attached to the compound.

In one embodiment the present disclosure provides a method of imaging myocardial perfusion comprising administering to a patient a contrast agent which comprises an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin, a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog; and scanning the patient using diagnostic imaging. In another embodment the imaging moiety is a radioisotope for nuclear medicine imaging, a paramagnetic species for use in MRI imaging, an echogenic entity for use in ultrasound imaging, a fluorescent entity for use in fluorescence imaging, or a light-active entity for use in optical imaging.

In another embodiment the present disclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben,

* pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog. In another embodiment the imaging moiety is a radioisotope for nuclear medicine imaging, a paramagnetic species for use in MRI imaging, an echogenic entity for use in ultrasound imaging, a fluorescent entity for use in fluorescence imaging, or a light- active entity for use in optical imaging.

In another embodiment the paramagnetic species for use in MRI imaging is

Gd", Fe**, In**, or Mn",

In another embodiment the echogenic entity for use in ultrasound imaging is a fluorocarbon encapsulated surfactant microsphere.

In another embodiment the radioisotope for nuclear medicine imaging is Hc, 13, 18F, 12, 127], ome 95Tc, ln, Cu, $C, "Ga, or ®Ga. In another embodiment the imaging moiety is 18F In another embodiment the imaging moiety is *¥™Tc.

In another embodiment the present diclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog wherein the contrast agent is of formula (I)

SEE

REN, A

R® RA , ““ a E §_A A

A : :

LO g Rr ! R14 c . A @, wherein each A is independently selected from O, CHR’, S, and NR’;

B is selected from hydrogen, Ci-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety;

C is selected from hydrogen, C1-Cs alkyl optionally substituted with an imaging moiety, an imaging moiety, and a bond to B;

D is selected from hydrogen, C,-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety;

E is selected from hydrogen, Ci-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; or

E and D, together with the carbon atom to which they are attached, form a double bond; or

E and D, together with the carbon atom to which they are attached, form a cyclopropyl ring;

Ta is a single or a double bond;

RL R%, R3, RY, RY, RR", and R™, are each independently selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety;

RS and R® are each independently selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, halo, hydroxy, and an imaging moiety; when present, R” and R® are independently selected from hydrogen, Ci-Cs alkyl optionally substituted with an imaging moiety, halo, hydroxy, and an imaging moiety; ox

RS and R’ together form an oxo group; or : RS and R® together form an oxo group; or

R” is O and R® is a bond to R’; provided that when "a is a double bond, R” and R® are absent;

R!! is hydrogen or hydroxy;

R12 is selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; or

R!! and R* together form an oxo group or =CHR'; with the proviso that at least one imaging moiety is present in formula (I).

In another embodiment

Ais O;

B and C are each independently CH; or CH,'®F;

D and E are each independently CHs or CH,'*F;

R}, RS, R’, and R'° are each independently hydrogen or 18%. and

R!! and R'? together form an oxo group.

In another embodiment the contrast agent is selected from ug “2 H z oF " Z 0 o oN ® y $0 : cho GY SRC

H 8 9 : o f ou , °~ ’ 6 JF ,

Z Zz 0 i 0 0 0 : 0 he Y 0. 0 _ fA . 17 H 2 ® @ H 0 o ® i ° ® H o 7 o_ ’ | o_ » i ou kd

Zz feocy oC

COC 9008 of or [o] H

[0] [e} [o)

N ~ ’ and T Og : ur

In another embodiment the present diclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog wherein the contrast agent is of formula (II),

G R24 ~N n

K

R28 /

Y. ~

L

R J

(ID, wherein 34

NS ¢

R22 ay) N N

SN

Ng R33 3

IN >

Vd

RZ S ~

SA Re R

Gis or R¥ , wherein misOorl; . a Jb. === and === gach independently represent a single ora double bond; rR”, R*, Rr, R*, R®, and R3* are independently selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; when present, R28 is selected from hydrogen and C;-Cs alkyl b optionally substituted with an imaging moiety, provided that when == is a double bond, R? is absent; : when present, R® is C;-Cg alkyl optionally substituted with an imaging moiety, provided that when =z is a double bond, R? is absent; 36 RY

R38

SE

Pis ® wherein R, R% RY, R®, and R¥ are independently selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; when present, P’ is hydrogen; or

P and P’ together form an 0Xo group; provided that when A is a double bond, P’ is absent;

Q is halo or haloalkyl;

Tis selected from NR?), 8, 0, C(=0), C(=0)0, NHCH,CH0, a bond, and

C(=0)N(R?"), with each group being drawn with its left end attached to G and its right end attached to the carbon substituted with R* and R%; when present, K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; when present, L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety;

M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or

L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring; nis 0, 1,2, 0r3; rR, RZ, RZ, R* RZ, and RS are independently selected from hydrogen, C;-

Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; and

Y is selected from a bond, carbon, and oxygen; provided that when Y is a bond, K and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, and heteroaryl; provided that at least one imaging moiety is present in formula (II).

In another embodiment R% is C;-Cg alkyl wherein the C.-C alkyl is tert- butyl.

In another embodiment R?® is C,-Cg alkyl wherein the C,-Cs alkyl is methyl.

In another embodiment the present diclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben,

pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog wherein the contrast agent is of formula (TID) 0

Q

RA RM KR rR?

A , R24 n rR? K

R% /

R25 SN

M

(1m), wherein:

J is selected from NR), S, 0, C(=0), C(=0)0, NHCH,CH,0, a bond, or

C(=0)N(R*"), with each group being drawn with its left end attached to G and its right end attached to the carbon substituted with R* and R%; when present, K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C1-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; when present, L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C1-Ce alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety;

M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C1-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or

L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring;

Q is halo or haloalkyl; nis 0, 1,2, or 3;

R* R%, R%,R%, R%, R%, and R? are independently selected from hydrogen,

C,-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety;

R is C;-Cs alkyl optionally substituted with an imaging moiety; and :

Y is selected from a bond, carbon, and oxygen; provided that when Y is a bond, K and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl, aryl, Ci-Cs alkyl optionally substituted with an imaging moiety, and heteroaryl; provided that at least one imaging moiety is present in formula (III).

In another embodiment J is O and R? is C,;-Cs alkyl wherein the C,-Cs alkyl is tert-butyl.

In another embodiment the contrast agent is selected from

Wy Sn geavN a

WE, BE and . -

In another embodiment the present diclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog wherein the contrast agent is of formula (IV):

RY ose

R® R35

R% Q

R2 R rR a BG \ R24

N J

/ n

R=

K

R* /

Y.

R? / SL

M avy, wherein:

J is selected from NR?"), S, 0, C(=0), C(=0)0, NHCH,CH;O, a bond, and

C(=O)NR?), with each group being drawn with its left end attached to G and its right end attached to the carbon substituted with R* and R%; when present, K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Ce alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety;

L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-C alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety;

M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-Cg alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or

L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring;

Q is halo or haloalkyl; nis 0,1, 2,0r 3; rR? RZ RZ, R% RS, R% RY, RZ, RS, R*, RY, R®, and R® are independently selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; and

Y is selected from a bond, carbon, and oxygen, provided that when Y isa bond, K and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl, aryl, C;-Cs alkyl optionally substituted with an imaging moiety, and heteroaryl; provided that at least one imaging moiety is present in formula (IV).

In another embodiment J is C(F0)N(H), and R* is Cy-Cs alkyl wherein the

C;-Cs alkyl is methyl.

In another embodiment the contrast agent is selected from , 1g

H H i ue : gen 1g - = \ ASUS »and 4 we

In another embodiment the present diclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog wherein the contrast agent is of formula (V)

RM

PN RY RZ

NY

T J n u K

R* / ~

Rr? J L

W), wherein

Tis selected from N(R?"), S, 0, C(=0), C(=0)0, NHCH;CH;0, 2 bond, and

CEONR™);

K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; when present, L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C1-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; : when present, M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C-

Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or

L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring;

T and U are independently selected from hydrogen, alkoxy, alkoxyalkyl, C;-

Cs alkyl optionally substituted with an imaging moiety, halo, and an imaging moiety; or

T and U, together with the carbon atoms to which they are attached, form a five- to six-membered aromatic or non-aromatic ring containing zero to two heterotoms selected from oxygen, nitrogen, and sulfur; wherein said ring is optionally substituted with one, two, or three substituents independently selected from C,-Ce alkyl optionally substituted with an imaging moiety and an imaging moiety; nis 0, 1,2, or 3; and rR? RZ, RZ, R* RY, R®, RY, and R* are independently selected from hydrogen, C1-Cs alky! optionally substituted with an imaging moiety, and an imaging moiety;

Y is selected from a bond, carbon, and oxygen, provided that when Yisa bond, X and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl, aryl, Ci-Ce alkyl optionally substituted with an imaging moiety, and heteroaryl; provided at least one imaging moiety is present in formula (V).

In another embodiment J is O.

In another embodiment the present diclosure provides a contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog wherein the contrast agent is of formula (VI)

R34

A R® ~N Re °~ R26 K

RS ML

(VD, wherein

RZ, R%, RZ, R%, and R* are independently selected from hydrogen, C-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; provided that at least one imaging moiety is present in formula (VI). -

In another embodiment the contrast agent is selected from

Neg Je 0 Neg o ~

PE p HOO

SN Ny ~n

SONNE >G EN GON wy , and .

Imaging moieties

Nuclear medicine contrast agents of the present disclosure include 'C, BN, 18p 123] 125] 99mTe SST, Hip 620y, $4Cu, Ga, and 68Ga. !'C Palmitate has been used to probe fatty acid oxidation and ''C-acetate has been used to assess oxidative metabolism in the myocardium (Circulation 1987, 76, 687-696). *N-Ammonia has been used widely to image myocardial perfusion (Circulation 1989, 80, 1328-37).

Agents based on '°F have been used as imaging agents for hypoxia and cancer (Drugs of the Future 2002, 27, 655-667). 15-(p-(‘*I)-iodophenyl)-pentadecanoic acid and 15-(p-("**T)-iodophenyl)-3(R,S)-methylpentadecanoic acid are two iodinated agents that have been used for imaging myocardial metabolism. In one embodiment, the imaging moiety employed in the present contrast agents is 18g Further imaging moieties of the present disclosure may be comprised of one or more X-ray absorbing or “heavy” atoms of atomic number 20 or greater, further comprising an optional linking moiety, L, between the parent molecular moiety and the X-ray absorbing atoms. A frequently used heavy atom in X-ray contrast agents is iodine. Recently,

X-ray contrast agents comprised of metal chelates (U.S. Pat. No. 5,417,959) and polychelates comprised of a plurality of metal ions (U.S. Pat. No. 5,679,810) have been disclosed. More recently, multinuclear cluster complexes have been disclosed as X-ray contrast agents (U.S. Pat. No. 5,804,161, WO 91/14460, and WO

92/17215). In certain embodiments of the present disclosure the specific metals used in the X-ray contrast agents include Re, Sm, Ho, Lu, Pm, Y, Bi, Pd, Gd, La, Au, Au,

Yb, Dy, Cu, Rh, Ag, and Ir.

MRI contrast agents of the present disclosure may be comprised of one or more analog moieties attached to one or more paramagnetic metal ions, further comprising an optional linking moiety, L, between the analog moieties and the paramagnetic metal ions. The paramagnetic metal ions may be present in the form of metal chelates or complexes or metal oxide particles. U.S. Pat. Nos. 5,412,148, and 5,760,191, describe examples of chelators for paramagnetic metal ions for use in

MRI contrast agents. U.S. Pat. No. 5,801,228, U.S. Pat. No. 5,567,411, and U.S. Pat.

No. 5,281,704, describe examples of polychelants useful for complexing more than one paramagnetic metal ion for use in MRI contrast agents. U.S. Pat. No. 5,520,904, describes particulate compositions comprised of paramagnetic metal ions for use as

MRI contrast agents. Examples of specific metals include G&**, Fe**, In**, and Mn".

The ultrasound contrast agents of the present disclosure may comprise a plurality of analog moieties attached to or incorporated into a microbubble of a biocompatible gas, a liquid carrier, and a surfactant microsphere, further comprising an optional linking moiety, L, between the analog moieties and the microbubble. In this context, the term “liquid carrier” means aqueous solution and the term wgurfactant” means any amphiphilic material which may produce a reduction in interfacial tension in a solution. A list of suitable surfactants for forming surfactant microspheres is disclosed, for example, in EP0727225A2. The term “surfactant microsphere” includes microspheres, nanospheres, liposomes, vesicles and the like.

The biocompatible gas can be any physiologically accepted gas, including, for example, air, or a fluorocarbon, such as a C3-Cs perfluoroalkane, which provides the difference in echogenicity and thus the contrast in ultrasound imaging. The gas may be encapsulated, contained, or otherwise constrained in or by the microsphere to which is attached the analog moiety, optionally via a linking group. The attachment can be covalent, ionic or by van der Waals forces. Specific examples of such contrast agents include, for example, lipid encapsulated perfluorocarbons with a plurality of tumor neovasculature receptor binding peptides, polypeptides or peptidomimetics.

Examples of gas filled imaging moieties include those found in U.S. Patent

Application Serial No. 09/931,317, filed August 16,2001, and U.S. Patent Nos.

5,088,499, 5,547,656, 5,228,446, 5,585,112, and 5,846,517.

Chelators

Many approaches to labeling compounds with IT are known, including direct labeling of the compound or inclusion of a chelating moiety (“chelator”). In one embodiment, the chelator is DADT, MAG3, MAMA, PAMA, or DOTA.

The compounds of the disclosure may optionally contain a chelator (“C”). In certain embodiments of the compounds of the disclosure, the chelator is a surfactant capable of forming an echogenic substance-filled lipid sphere or microbubble. In certain other embodiments, the chelator is a bonding unit having a formula selected from

Al /

FA B

Al PN 2 1

A a2 Ng —E E E A?

Al ~~” ~~ ~ Sg? ~~ E

J:

Nez

E E E

Al” Na” ~~ Nal

E E E E

Rd Nai” ~a37 Na al \ Ng SE 4 Al Al

Al {

M3—E E—A! / NY

Jk !

Al {

E—A \

E

/

Al , and

F AS

A wherein each A! is independently selected from _NR*RY, .NHR®, -SH, -S(Pg), -OH,

PR¥RY", -P(O)R*R*, and a bond to the compound that binds MC-1; each A? is independently selected from NER), NR*), S, 0, PR*), and -OP(O)R*)0-;

Adis N;

A% is selected from OH and OC(=0)C1-Czo alkyl;

AS is OC(=0) C1-Cyo alkyl; each E is independently selected from C;-Cj¢ alkylene substituted with 0-3

R30, C¢-Cjo arylene substituted with 0-3 R%, C3-C)p cycloalkylene substituted with 0- 3 R®, heterocyclyl-Ci-Cio alkylene substituted with 0-3 R%, C¢-Cyo aryl-Ci-Cro alkylene substituted with 0-3 R*, and heterocyclylene substituted with 0-3 R*;

E! is selected from a bond and E; each E2 is independently selected from C;-Ce alkyl substituted with 0-3 R,

Ce-Cio aryl substituted with 0-3 R*’, C3-Cyo cycloalkyl substituted with 0-3 R*, heterocyclyl-Ci-C)o alkyl substituted with 0-3 RC, Cs-Cyo aryl-Ci-Cyp alkyl substituted with 0-3 R®, C;-Co alkyl-Ce-Cio aryl substituted with 0-3 RC, and heterocyclyl substituted with 0-3 R®’;

E? is C;-Cjo alkylene substituted with 1-3 R*”;

Pg is a thiol protecting group;

R* and R* are each independently selected from a bond to the compound that binds MC-1, hydrogen, C1-Cy alkyl substituted with 0-3 R*, aryl substituted with 0-3 R®, C3-Cyo cycloalkyl substituted with 0-3 R¥, heterocyclyl-Ci-Cio alkyl substituted with 0-3 R*®, Cg-Cyo aryl-Ci-Cio alkyl substituted with 0-3 R%, and heterocyclyl substituted with 0-3R%;

R*® and R* are each independently selected from a bond to the compound that binds MC-1, -OH, C1-Cio alkyl substituted with 0-3 R®, aryl substituted with 0-3

RR, C3-C) cycloalkyl substituted with 0-3 R¥®, heterocyclyl-Ci-Cio alkyl substituted with 0-3 R%, Cs-Cyo aryl-Cy-Cyo alkyl substituted with 0-3 R% and heterocyclyl substituted with 0-3 R*; each R* is independently selected from a bond to the compound that binds

MC-1, =O, halo, trifluoromethyl, cyano, -COR", -C(=O)R®, -C(=0)NR*")z, -CHO, -CH,0R"!, _OC(=0)R®", -OC(=0)OR*, -OR*', -OC(=OINR"") NR®*'C(=O)R*, -

NR*'C(=0)OR”,

NRS'C(=O)N(R®!),, NRSONR 2, NR'SO,R?!, -SO3H, -SOR”, _SR®, -S(=O)R®", -SONR""), N(R), _NHC(=S)NHR®!, =NOR*!, NO, -

C(=O)NHOR’!, -C(F0)NHN(R*")2, -OCH,CO-H, 2-(1-morpholino)ethoxy, C1-Cs alkyl, C,-C, alkenyl, C3-Cs cycloalkyl, C3-Cs cycloalkylmethyl, C;-Ce alkoxyalkyl, aryl substituted with 0-2 R*', and heterocyclyl; each R%! is independently selected from a bond to the compound that binds

MC-1, hydrogen, C-Cs alkyl, phenyl, benzyl, and Cy. alkoxy;

R® is a co-ordinate bond to a metal; each R® selected from R®, =0, -COR®, -C(=O)R®, -C(=O)NR*), -

CH,OR®, -OR®, N(R®*),, and C-C alkenyl; each R¥ is independently selected from R®!, hydrogen, C;-Cs alkyl, phenyl, benzyl, and trifluoromethyl; and

R®! is a bond to the compound that binds MC-1; wherein at least one of A}, R%, RY, R*, R*’, R®, R*', and R® is a bond to the compound that binds MC-1.

Methods of Making

Typically °F labeled compounds are synthesized by S,2 displacement of an appropriate leaving group. These leaving groups are preferrably sulfonic acid esters such as toluenesulfonate (tosylate, TsO), methanesulfonate (mesylate, MsO), or trifluoromethanesulfonate (triflate, TO). The leaving group may also be a halide, a phosphineoxide (via Mitsunobu reaction), or an internal leaving group (such as an epoxide or cyclic sulfate). These compounds are made from highly activated, dry

K'®F, that is made “hotter” by the addition of cryptands such as krytofix(2.2.2}.

Purification is generally via salt removal by reverse-phase chromatography (Sep-

Pak).

Representative methods of making the contrast agents are described in the following examples. The foregoing chemical transformations may be conducted using techniques which would be readily apparent to one of ordinary skill in the art, once armed with the teachings in the present applications. Representative reaction : solvents include, for example, DMF, NMP, DMSO, THF, ethyl acetate, dichloromethane, and chloroform. The reaction solution may be kept neutral or basic by the addition of an amine such as triethylamine or DIEA. Reactions may be carried out at ambient temperatures and protected from oxygen and water with a nitrogen atmosphere.

Temporary protecting groups may be used to prevent other reactive functionality, such as amines, thiols, alcohols, phenols, and carboxylic acids, from participating in the reaction. Representative amine protecting groups include, for example, tert-butoxycarbonyl and trityl (removed under mild acidic conditions),

Fmoc (removed by the use of secondary amines such as piperidine), and benzyloxycarbonyl (removed by strong acid or by catalytic hydrogenolysis). The ‘trityl group may also used for the protection of thiols, phenols, and alcohols. In certain embodiments the carboxylic acid protecting groups include, for example, tert- butyl ester (removed by mild acid), benzyl ester (usually removed by catalytic hydrogenolysis), and alkyl esters such as methyl or ethyl (usually removed by mild base). All protecting groups may be removed at the conclusion of synthesis using the conditions described above for the individual protecting groups, and the final product may be purified by techniques which would be readily apparent to one of ordinary skill in the art, once armed with the present disclosure.

Use

The contrast agents of the present disclosure may be used in a method of imaging, including methods of imaging in a patient comprising administering the contrast agent to the patient by injection, infusion, or any other known method, and imaging the area of the patient wherein the event of interest is located.

The useful dosage to be administered and the particular mode of administration will vary depending upon such factors as age, weight, and particular region to be treated, as well as the particular contrast agent used, the diagnostic use contemplated, and the form of the formulation, for example, suspension, emulsion, microsphere, liposome, or the like, as will be readily apparent to those skilled in the art.

Typically, dosage is administered at lower levels and increased until the desirable diagnostic effect is achieved. In one embodiment, the above-described contrast agents may be administered by intravenous injection, usually in saline solution, at a.dose of about 0.1 to about 100 mCi per 70 kg body weight (and all combinations and subcombinations of dosage ranges and specific dosages therein), or preferably at a dose of about 0.5 to about 50 mCi. Imaging is performed using techniques well known to the ordinarily skilled artisan.

For use as nuclear medicine contrast agents, the compositions of the present disclosure, dosages, administered by intravenous injection, will typically range from about 0.5 umol/kg to about 1.5 mmol/kg (and all combinations and subcombinations of dosage ranges and specific dosages therein), preferably about 0.8 pmol/kg to about 1.2 mmol/kg.

For use as MRI contrast agents, the compositions of the present disclosure : may be used in a similar manner as other MRI agents as described in U.S. Patent No. 5,155,215; U.S. Patent No. 5,087,440; Magn. Reson. Med. 1986, 3, 808; Radiology 1988, 166, 835; and Radiology 1988, 166, 693. Generally, sterile aqueous solutions of the contrast agents may be administered to a patient intravenously in dosages ranging from about 0.01 to about 1.0 mmoles per kg body weight (and all combinations and subcombinations of dosage ranges and specific dosages therein).

The ultrasound contrast agents of the present disclosure may be administered by intravenous injection in an amount from about 10 to about 30 pL (and all combinations and subcombinations of dosage ranges and specific dosages therein) of the echogenic gas per kg body weight or by infusion at a rate of approximately 3 nL/kg/min.

Another aspect of the present disclosure is diagnostic kits for the preparation of diagnostic agents for detecting, imaging, and/or monitoring myocardial perfusion.

Diagnostic kits of the present disclosure comprise one or more vials containing the sterile, non-pyrogenic, formulation comprising a predetermined amount of a reagent of the present disclosure, and optionally other components such as one or two ancillary ligands such as tricine and 3-[bis(3-sulfophenyl)phosphine]benzenesulfonic acid (TPPTS), reducing agents, transfer ligands, buffers, lyophilization aids, stabilization aids, solubilization aids and bacteriostats. The kits may also comprise a reducing agent, such as, for example, tin(T{).

Buffers useful in the preparation of contrast agents and kits include, for example, phosphate, citrate, sulfosalicylate, and acetate buffers. A more complete list can be found in the United States Pharmacopoeia.

Lyophilization aids useful in the preparation of contrast agents and kits include, for example, mannitol, lactose, sorbitol, dextran, FICOLL® polymer, and polyvinylpyrrolidine (PVP).

Stabilization aids useful in the preparation of contrast agents and kits include, for example, ascorbic acid, cysteine, monothioglycerol, sodium bisulfite, sodium metabisulfite, gentisic acid, and inositol.

Solubilization aids useful in the preparation of contrast agents and kits include, for example, ethanol, glycerin, polyethylene glycol, propylene glycol, polyoxyethylene sorbitan monooleate, sorbitan monoloeate, polysorbates, poly(oxyethylene)-poly(oxypropylene)-poly(oxyethylene) block copolymers (“Pluronics”) and lecithin. In certain embodiments the solubilizing aids are polyethylene glycol and Pluronics.

Bacteriostats useful in the preparation of contrast agents and kits include, for example, benzyl alcohol, benzalkonium chloride, chlorbutanol, and methyl, propyl, or butyl paraben.

A component in a diagnostic kit can also serve more than one function. For example, a reducing agent for a radionuclide can also serve as a stabilization aid, or a buffer can also serve as a transfer ligand, or a lyophilization aid can also serve as a transfer, ancillary, or co-ligand.

The compounds herein described may have asymmetric centers. Unless otherwise indicated, all chiral, diastereomeric and racemic forms are included in the present disclosure. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. It will be appreciated that compounds of the present disclosure may contain asymmetrically substituted carbon atoms, and may be isolated in optically active or racemic forms. It is well known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis from optically active starting materials. Two distinct isomers (cis and trans) of the peptide bond are known to occur; both can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. The D- and L-isomers of a particular amino acid are designated herein using the conventional 3-letter abbreviation of the amino acid, as indicated by the following examples: D-Leu, or L-Leu.

For the sake of simplicity, connection points (“-”") are not depicted. When an atom or compound is described to define a variable, it is understood that it is intended to replace the variable in a manner to satisfy the valency of the atom or compound.

For example, if a variable A” was identified as CR¥®=CRY), both carbon atoms would form a part of the chain in order to satisfy their respective valences.

When any variable occurs more than one time in any substituent or in any formula, its definition in each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group, or plurality of groups, is shown to be substituted with 0-2 R®, then said group(s) may optionally be substituted withup to two R®, and R at each occurrence in each group is selected independently from the defined list of possible R¥. Also, by way of example, for the group NR), each of the two R®! substituents on N is independently selected from the defined list of possible R®! Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. When a bond to a substituent is shown to cross the bond connecting two atoms in a ring, then such substituent may be bonded to any atom on the ring.

Definitions

The number of carbon atoms in any particular group is denoted before the recitation of the group. For example, the term “Cg-C aryl” denotes an aryl group containing from six to ten carbon atoms, and the term «Cg-Croaryl-Ci-Croalkyl,” refers to an aryl group of six to ten carbon atoms attached to the parent molecular moiety through an alkyl group of one to ten carbon atoms.

The term “alkenyl,” as used herein, refers to a straight or branched chain hydrocarbon containing at Jeast one carbon-carbon double bond.

The term “alkoxy,” as used herein, refers to a C;-Cs alkyl group attached to the parent molecular moiety through an oxygen atom.

The term “alkoxyalkyl,” as used herein, refers to a C;-Cs alkyl group substituted with one, two, or three alkoxy groups.

The term “alkyl,” as used herein, refers to a group derived from a straight or branched chain saturated hydrocarbon.

The term “alkylaryl,” as used herein, refers to an alkyl group attached to the parent molecular moiety through an aryl group.

The term “alkylene,” as used herein, refers to a divalent group derived from a straight or branched chain saturated hydrocarbon.

The term “alkyloxy,” as used herein, refers to a C;-Ce alkyl group attached to the parent molecular moiety through an oxygen atom.

The term “analog moiety,” as used herein, refers to the compounds of the present disclosure excluding the imaging moiety or moieties.

The term “aryl,” as used herein, refers to a phenyl group, or a bicyclic fused ring system wherein one or more of the rings is a phenyl group. Bicyclic fused ring systems consist of a phenyl group fused to a monocyclic cycloalkenyl group, a monocyclic cycloalkyl group, or another phenyl group. The aryl groups of the present invention can be attached to the parent molecular moiety through any substitutable carbon atom in the group. Representative examples of aryl groups include, but are not limited to, anthracenyl, azulenyl, fluorenyl, indanyl, indenyl, naphthyl, phenyl, and tetrahydronaphthyl.

The term “arylalkyl,” as used herein, refers to an alkyl group substituted with one, two, or three aryl groups.

The term “arylalkylene,” as used herein, refers to a divalent arylalkyl group, where one point of attachment to the parent molecular moiety is on the aryl portion and the other is on the alkyl portion.

The term “arylene,” as used herein, refers to a divalent aryl group.

As used herein, the terms “ancillary” or “co-ligands” refers to ligands that serve to complete the coordination sphere of the radionuclide together with the chelator or radionuclide bonding unit of the reagent. For radiopharmaceuticals comprising a binary ligand system, the radionuclide coordination sphere comprises one or more chelators or bonding units from one or more reagents and one or more ancillary or co-ligands, provided that there are a total of two types of ligands, chelators or bonding units. For example, a radiopharmaceutical comprised of one chelator or bonding unit from one reagent and two of the same ancillary or co-ligands and a radiopharmaceutical comprising two chelators or bonding units from one or two reagents and one ancillary or co-ligand are both considered to comprise binary ligand systems. For radiopbarmaceuticals comprising a ternary ligand system, the radionuclide coordination sphere comprises one or more chelators or bonding units from one or more reagents and one or more of two different types of ancillary or co-ligands, provided that there are a total of three types of ligands, chelators or bonding units. For example, a radiopharmaceutical comprised of one chelator or bonding unit from one reagent and two different ancillary or co-ligands is considered to comprise a ternary ligand system.

Ancillary or co-ligands useful in the preparation of radiopharmaceuticals and in diagnostic kits useful for the preparation of said radiopharmaceuticals comprise one or more oxygen, nitrogen, carbon, sulfur, phosphorus, arsenic, selenium, and tellurium donor atoms. A ligand canbe a transfer ligand in the synthesis of a radiopharmaceutical and also serve as an ancillary or co-ligand in another radiopharmaceutical. Whether a ligand is termed a transfer or ancillary or co-ligand depends on whether the ligand remains in the radionuclide coordination sphere in the radiopharmaceutical, which is determined by the coordination chemistry of the radionuclide and the chelator or bonding unit of the reagent or reagents.

A “bacteriostat” is a component that inhibits the growth of bacteria in a formulation either during its storage before use of after a diagnostic kit is used to synthesize a radiopharmaceutical.

The term “bubbles” or “microbubbles,” as used herein, refers to vesicles which are generally characterized by the presence of one or more membranes or walls surrounding an internal void that is filled with a gas or precursor thereto.

Exemplary bubbles or microbubbles include, for example, liposomes, micelles, and the like.

The terms “chelator” and “bonding unit,” as used herein, refer to the moiety or group on a reagent that binds to a metal ion through one or more donor ators.

The term “contrast agent,” as used herein, refers to an agent used to highlight specific areas so that organs, blood vessels, and/or tissues are more visible. By increasing the visibility of the surfaces being studied, the presence and extent of disease and/or injury can be determined.

The term “cycloalkenyl,” as used herein, refers to a non-aromatic, partially unsaturated monocyclic, bicyclic, or tricyclic ring system having three to fourteen carbon atoms and zero heteroatoms. Representative examples of cycloalkenyl groups include, but are not limited to, cyclohexenyl, octahydronaphthalenyl, and norbornylenyl.

The term “cycloalkyl,” as used herein, refers to a saturated monocyclic, bicyclic, or tricyclic hydrocarbon ring system having three to fourteen carbon atoms and zero heteroatoms. Representative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclopentyl, bicyclo[3.1.1]heptyl, and adamantyl.

The term “C3-Cjo cycloalkylene,” as used herein, refers to a divalent cycloalkyl group containing from three to ten carbon atoms.

The term “diagnostic imaging,” as used herein, refers to a procedure used to detect a contrast agent.

A “diagnostic kit” or “kit” comprises a collection of components, termed the formulation, in one or more vials which are used by the practicing end user ina clinical or pharmacy setting to synthesize diagnostic radiopharmaceuticals. The kit preferably provides all the requisite components to synthesize and use the diagnostic pharmaceutical except those that are commonly available to the practicing end user, such as water or saline for injection, a solution of the radionuclide, equipment for heating the kit during the synthesis of the radiopharmaceutical, if required, equipment necessary for administering the radiopharmaceutical to the patient such as syringes, shielding, imaging equipment, and the like. Contrast agents are provided to the end user in their final form in a formulation contained typically in one vial, as either a lyophilized solid or an aqueous solution. The end user typically reconstitutes the lyophilized material with water or saline and withdraws the patient dose or just withdraws the dose from the aqueous solution formulation as provided.

The term “donor atom,” as used herein, refers to the atom directly attached to a metal by a chemical bond.

The terms “halo” and “halogen,” as used herein, refer to F, Cl, Br, or I.

The term “haloalkyl,” as used herein, refers to a C,-Cs alkyl group substituted by one, two, three, or four halogen atoms.

The term “heteroaryl,” as used herein, refers to an aromatic five- or six- membered ring where at least one atom is selected from N, O, and S, and the remaining atoms are carbon. The term “heteroaryl” also includes bicyclic systems where a heteroaryl ring is fused to a four- to six-membered aromatic or non-aromatic ring containing zero, one, or two additional heteroatoms selected from N, O, and S.

The heteroaryl groups are attached to the parent molecular moiety through any substitutable carbon or nitrogen atom in the group. Representative examples of heteroaryl groups include, but are not limited to, benzoxadiazolyl, benzoxazolyl, benzofuranyl, benzothienyl, furanyl, imidazolyl, indazolyl, indolyl, isoxazolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, quinolinyl, thiazolyl, thienopyridinyl, thienyl, triazolyl, thiadiazolyl, and triazinyl.

The term “heterocyclyl,” as used herein, refers to a five-, six-, or seven- membered ring containing one, two, or three heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. The five-membered ring has zero to two double bonds and the six- and seven-membered rings have zero to three double bonds. The term “heterocyclyl” also includes bicyclic groups in which the heterocyclyl ring is fused to a phenyl group, a monocyclic cycloalkenyl group, a monocyclic cycloalkyl group, or another monocyclic heterocyclyl group. The heterocyclyl groups of the present invention can be attached to the parent molecular moiety through a carbon atom or a nitrogen atom in the group. Examples of heterocyclyl groups include, but are not limited to, benzothienyl, furyl, imidazolyl, indolinyl, indolyl, isothiazolyl, isoxazolyl, morpholinyl, oxazolyl, piperazinyl, piperidinyl, pyrazolyl, pyridinyl, pyrrolidinyl, pyrrolopyridinyl, pyrrolyl, thiazolyl, thienyl, and thiomorpholinyl.

The term “heterocyclylalkyl,” as used herein, refers to an alkyl group substituted with one, two, or three heterocyclyl groups.

The term “heterocyclylalkylene,” as used herein, refers to a divalent heterocyclylalkyl group, where one point of attachment to the parent molecular moiety is on the heterocyclyl portion and the other is on the alkyl portion.

The term “heterocyclylene,” as used herein, refers to a divalent heterocyclyl group.

The term “hydroxy,” as used herein, refers to —OH.

The term “imaging moiety,” as used herein, refer to a portion or portions ofa molecule that allow for the detection, imaging, and/or monitoring of the presence and/or progression of a condition(s), pathological disorder(s), and/or disease(s).

The term “linking group,” as used herein, refers to a portion of a molecule that serves as a spacer between two other portions of the molecule. Linking groups may also serve other functions as described herein. Examples of linking groups include linear, branched, or cyclic alkyl, aryl, ether, polyhydroxy, polyether, polyamine, heterocyclic, aromatic, hydrazide, peptide, peptoid, or other physiologically compatible covalent linkages or combinations thereof.

As used herein, the term “lipid” refers to a synthetic or naturally-occurring amphipathic compound which comprises a hydrophilic component and a hydrophobic component. Lipids include, for example, fatty acids, neutral fats, phosphatides, glycolipids, aliphatic alcohols and waxes, terpenes and steroids. Exemplary compositions which comprise a lipid compound include suspensions, emulsions and vesicular compositions. “Liposome” refers to a generally spherical cluster or aggregate of amphipathic compounds, including lipid compounds, typically in the form of one or more concentric layers, for example, bilayers. They may also be referred to herein as lipid vesicles.

A “lyophilization aid” is a component that has favorable physical properties for lyophilization, such as the glass transition temperature, and is generally added to the formulation to improve the physical properties of the combination of all the components of the formulation for lyophilization.

The term “oxo,” as used herein, refers to =0.

As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term “pharmaceutically acceptable salt,” as used herein, represents salts or zwitterionic forms of the compounds of the present invention which are water or oil-soluble or dispersible, which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio, and are effective for their intended use The salts can be prepared during the final isolation and purification of the compounds or separately by reacting a suitable nitrogen atom with a suitable acid. Representative acid addition salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate; digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, npicotinate, 2-naphthalenesulfonate, oxalate, palmoate, pectinate, persulfate, 3-phenyliproprionate, picrate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, para-toluenesulfonate, and undecanoate. Examples of acids which can be employed to form pharmaceutically acceptable addition salts include inorganic acids such as hydrochloric, hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic, maleic, succinic, and citric.

By “reagent” is meant a compound of this disclosure capable of direct transformation into a metallopharmaceutical of this disclosure. Reagents may be utilized directly for the preparation of the metallopharmaceuticals of this disclosure or may be a component in a kit of this disclosure.

A “reducing agent” is a compound that reacts with a radionuclide, which is typically obtained as a relatively unreactive, high oxidation state compound, to lower its oxidation state by transferring electron(s) to the radionuclide, thereby making it more reactive. Reducing agents useful in the preparation of radiopharmaceuticals and in diagnostic kits useful for the preparation of said radiopharmaceuticals include, for example, stannous chloride, stannous fluoride, formamidine sulfinic acid, ascorbic acid, cysteine, phosphines, and cuprous or ferrous salts. Other reducing agents are described, for example, in Brodack et. al., PCT Application 94/22496.

A “stabilization aid” is a component that is typically added to the metallopharmaceutical or to the diagnostic kit either to stabilize the metallopharmaceutical or to prolong the shelf-life of the kit before it must be used.

Stabilization aids can be antioxidants, reducing agents or radical scavengers and can provide improved stability by reacting preferentially with species that degrade other components or the metallopharmaceuticals.

By “stable compound” or “stable structure” is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious pharmaceutical agent.

A “solubilization aid” is a component that improves the solubility of one or more other components in the medium required for the formulation.

The term “thiol protecting group,” as used herein, refers to a group intended to protect a thiol group against undesirable reactions during synthetic procedures.

Any thiol protecting group known in the art may be used. Examples of thiol protecting groups include, but are not limited to, the following: acetamidomethyl, benzamidomethyl, 1-ethoxyethyl, benzoyl, and triphenylmethyl.

A “transfer ligand” is a ligand that forms an intermediate complex with a metal ion that is stable enough to prevent unwanted side-reactions but labile enough to be converted to a contrast agent. The formation of the intermediate complex is kinetically favored while the formation of the metallopharmaceutical is thermodynamically favored. Transfer ligands useful in the preparation of contrast agents and in diagnostic kits useful for the preparation of diagnostic radiopharmaceuticals include, for example, gluconate, glucoheptonate, mannitol, glucarate, N,N,N’ N’-ethylenediaminetetraacetic acid, pyrophosphate and methylenediphosphonate. In general, transfer ligands are comprised of oxygen or nitrogen donor atoms.

As used herein, the term ‘“‘vesicle” refers to a spherical entity which is characterized by the presence of an internal void. In one embodiment vesicles are formulated from lipids, including the various lipids described herein. In any given vesicle, the lipids may be in the form of a monolayer or bilayer, and the mono- or bilayer lipids may be used to form one of more mono- or bilayers. In the case of more than one mono- or bilayer, the mono- or bilayers are generally concentric. The lipid vesicles described herein include such entities commonly referred to as liposomes, micelles, bubbles, microbubbles, microspheres and the like. Thus, the lipids may be used to form 2 unilamellar vesicle (comprised of one monolayer or bilayer), an oligolamellar vesicle (comprised of about two or about three monolayers or bilayers) or a multilamellar vesicle (comprised of more than about three monolayers or bilayers). The internal void of the vesicles may be filled with a liquid, including, for example, an aqueous liquid, a gas, a gaseous precursor, and/or a solid or solute material, including, for example, a bioactive agent, as desired.

As used herein, the term “vesicular composition” refers to a composition which is formulate from lipids and which comprises vesicles.

The present disclosure will now be described in connection with certain embodiments which are not intended to limit its scope. On the contrary, the present disclosure covers all alternatives, modifications, and equivalents as can be included within the scope of the claims. Thus, the following examples will illustrate one practice of the present invention, it being understood that the examples are for the purposes of illustration of certain embodiments and are presented to provide what is believed to be the most useful and readily understood description of its procedures and conceptual aspects. :

Synthesis of Fenazaquin Analog:

Example 1A

Synthesis of 4-[4-2-Hydroxyethyl)phenyl]-4-oxo-butyric acid methyl ester h Jon cl CO,Me

OH bow i aE oe 2. Sodium HO

MeOH

To a dry 250 mL flask under a nitrogen atmosphere was added phenethyl alcohol (2.50 g, 0.02 mol), anhydrous dichloromethane (150 mL), and methyl-4- chloro-4-oxobutyrate (6.02 g, 0.04 mol). The contents of the flask were cooled to 0 °C with an ice bath. To the solution was added aluminum chloride (25 g, 0.2 mol) in portions being careful to avoid a violent exotherm. The resulting yellowish mixture was stirred for 3 hours. At this point the reaction was quenched with ice water. The mixture was diluted with dichloromethane and transferred to a separatory funnel.

The organic layer was washed with a saturated solution of sodium bicarbonate, brine and then dried over magnesium sulfate. Filtration and concentration of the filtrate under reduced pressure provided a crude yellow oil. The oil was suspended in anhydrous methanol (100 mL) and sodium metal was added to the mixture until a pH of 9 was obtained. The mixture was stirred for 3 hours. The volume was reduced and then diluted with ethyl acetate. The solution was transferred to a separatory funnel and washed with aqueous 0.05 N hydrochloric acid, brine and dried over magnesium sulfate. The solution was concentrated under reduced pressure to give a crude yellow oil with a mass of 5.88 g. Column chromatography [silica gel; eluent hexanes-ethyl acetate (3:2)] provided the desired product (2.69 g, 57 %). 'H (CDCl3) 8(ppm): 2.65 (t, 2H); 2.81 (t, 2H); 3.19 (t, 2H); 3.6 (s, 3H); 3.75 (1, 2H); 7.22 (4, 2H); 7.81 (d, 2H). 13¢C (CDCl) 8(ppm): 27.76, 33.03, 38.66, 51.52, 62.68, 127.97, 128.99, 134.47, 144.78, 173.21,197.64. - Example 1B : Synthesis of 4-[4-(2-hydroxyethyDphenyl]butyric acid methyl ester fo) Hy, Pd/C

Neoas come _ MeOH ohe CO Me

HO HO

A mixture of Example 1A (2.50 g, 11 mmol), 10 % Pd/C (0.25 g, 0.23 mmol of Pd metal) in anhydrous methanol (25 mL) was first degassed to remove air (two vacuum/H, cycles) after which it was capped and a balloon filled with H, was applied to it for 12 hours. After this time the reaction mixture was filtered through diatomaceous earth (Celite®) and the filtrate was concentrated under reduced pressure to give 2.32 g of crude material. Column chromato graphy [silica gel; eluent hexanes-cthyl acetate (2:1)] provided the desired product (0.92 g, 39 %). 'H (CDCl3) (ppm): 1.91-1.96 (m, 2H); 2.32 (t, 2H); 2.62 (t, 2H); 2.83 (t, 2H); 3.66 (s, 3H); 3.85 (t, 2H); 7.11-7.15 (m, 4H).

Example 1C

Synthesis of 4-{4-[2-(quinazolin-4-yloxy)ethyllphenyl}butyric acid methyl ester

CO,Me cl NaH

HO N AN

CL

A dry 50 mL flask was fitted with an addition funnel. To the flask were added 4-chloroquinazoline (592 mg, 3.6 mmol), anhydrous tetrahydrofuran (10 mL), and 60 wt % sodium hydride (187 mg, 4.7 mmol). A solution of Example 1B (800 mg, 3.6 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise using the addition funnel. The reaction was stirred for 3.5 hours. The reaction was diluted with ethyl acetate and quenched by the addition of aqueous 0.1 N hydrochloric acid.

The mixture was transferred to a separatory funnel and washed with brine. The organic layer was dried over magnesium sulfate, filtered, and concentrated. Column chromatography [silica gel; eluent hexanes-ethyl acetate (4:1)] provided the desired product (538 mg, 43 %). 'H(CDCl,) 8(ppm): 1.92-1.98 (m, 2H); 2.33 (t, 2H); 2.64 (1, 2H); 3.19 (t, 2H); 3.66 (s, 3H); 4.79 (t, 2H); 7.15 (d, 2H); 7.27 (4, 2H); 7.57 (t, 1H); 7.83 (t, 1H); 7.94 (d, 1H); 8.15 (d, 1H); 8.80 (s, 1H). 26.68, 33.59, 34.93, 35.03, 51.67, 67.89, 116.48, 123.72, 127.23, 127.82, 128.87, 129.24, 133.74, 135.76, 139.90, 151.08, 154.56, 166.89, 174.10.

Example 1D

Synthesis of 4-{4-[2-(Quinazolin4-yloxy)ethyl]phenyl}butan-1-ol ~~ COMe 1 (AH a x ether xX 2. MnO; 0 DCM lo) =N =N

To a dry 15 mL flask was added lithium aluminum hydride (233 mg, 6.0 mmol) and anhydrous diethyl ether (3 mL). The mixture was cooled with an ice bath.

A solution of Example 1C (538 mg, 1.54 mmol) in anhydrous diethyl ether (3 mL) was slowly added with vigorous stirring. The bath was removed and the slurry was stirred for 15 minutes. The reaction was quenched with water (0.233 mL), aqueous

% sodium hydroxide (0.233 mL) and water (0.699 mL). The white solid was filtered and the filtrate was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give a clear oil. The oil was then dissolved in anhydrous dichloromethane (10 mL) and manganese(TV) oxide (500 mg, 5.8 mmol) was added to the solution. The mixture was stirred for 12 hours. Filtration through diatomaceous earth (Celite®) followed by concentration of the filtrate under reduced pressure afforded 395 mg of crude product. Column chromatography [silica gel; eluent pentane-ethyl acetate (2:3)] provided the desired product (225 mg, 49 %). 'H (CDCl3) 8(ppm): 1.55-1.61 (m, 2H); 1.65-1.68 (m, 2H); 2.61 (t, 2H); 3.17 (t, 2H); 3.64 (t, 2H); 4.79 (t, 2H); 7.12 (d, 2H); 7.23 (d, 2H); 7.56 (t, 1H); 7.82 (t, 1H); 7.93 (d, 1H); 8.14 (d, 1H); 8.77 (5, 1H). °C (CDCly) (ppm): 27.52, 32.31, 34.89, 35.21, 62.81, 67.74, 116.67, 123.54, 127.08, 127.49, 128.63, 128.98, 133.61, 135.23, 140.64, 150.68, 154.29, 166.79.

Example 1E

Synthesis of Toluene-4-sulfonic acid 4-{4-[2«(quinazolin-4- yloxyethyl]phenyl}butyl ester

OH OTs

Poa wo Poin

CL CY

Ne Ne

To a dry 10 mL flask was added p-toluenesulfonyl chloride (32.5 mg, 0.17 mmol), 4-(dimethylamino)pyridine (20.7 mg, 0.17 mmol), Example 1D (50.0 mg, 0.16 mmol), anhydrous dichloromethane (1 mL) and triethylamine (17.2 mg, 0.17 mmol). The resulting solution was stirred for 2 hours, concentrated under reduced pressure, and purified by column chromatography [silica gel; eluent pentane-ethyl acetate (1.86:1)] to provide the desired product (52 mg, 70 %). 'H(CDCl3) §(ppm): 1.64-1.68 (m, 4H); 2.44 (s, 3H); 2.56 (t, 2H); 3.19 (t, 2H); 4.04 (t, 2H); 4.78 (t, 2H); 7.08 (d, 2H); 7.26 (d, 2H); 7.57 (t, 1H); 7.78 (d, 2H); 7.84 (t, 1H), 8.14 (d, 1H); 8.80 (s, 1H).

Example 1F

Synthesis of 4-{2-[4-(4-Fluorobutyl)phenyl] ethoxy}quinazoline

OTs F

Poa . Pan lo) Kryptofix 0}

N SN

A dry 5 mL flask was fitted with a reflux condenser. To the flask was added potassium fluoride (6.1 mg, 0.1 mmol), kryptofix (40 mg, 0.1 mmol) and anhydrous acetonitrile (0.5 mL). To the resulting solution was added a solution of Example 1E (25 mg, 0.05 mmol) in anhydrous acetonitrile (1 mL). The flask was placed in a 90 oC oil bath. The solution was stirred for 1 hour. After cooling the reaction mixture was diluted with diethyl ether, transferred to a separatory funnel, and washed with aqueous 0.1 N hydrochloric acid, saturated aqueous solution of sodium bicarbonate, and then brine. The organic layer was dried with magnesium sulfate, filtered, and concentrated under reduced pressure. Column chromatography [silica gel; eluent hexanes-ethyl acetate (3:1)] provided the desired product (10.7 mg, 63 %).

IH(CDCls) 8(ppm): 1.65-1.73 (m, 4H); 2.63 (t, 2H); 3.17 (t, 2H); 4.40 (t, 1H); 4.48 (t, 1H); 4.77 (t, 2H); 7.13 (d, 2H); 7.24 (d, 2H); 7.55 (1H); 7.82 (t, 1H); 7.92 (d, 1H); 8.13 (d, 1H); 8.78 (s, 1H). "°C (CDCL) 8(ppm): 27.19 (d, Jor = 4.5), 30.20 (d, Jer =19.5), 35.15 (d, 2Jcr = 27.0), 67.94, 84.17 (4, or = 163.3), 116.93, 123.75, 127.26, 127.84, 128.82, 129.23, 129.42, 133.77, 135.62, 138.21, 140.54, 151.08, 154.59.

I9%(CDCl, CFCl internal standard) (ppm): -218.59 (tof t, J = -27.6, -50.4).

Synthesis of Pyridaben Analogs:

Example 2A

Synthesis of Butyric acid 4-phenylbutyl ester 0 ohas Te Shand

DCM 0

To 4-phenyl-1-butanol (7.0 g, 0.047 mol) was added anhydrous dichloromethane (20 mL). A solution of butyryl chloride (4.79 g, 0.045 mol) in anhydrous dichloromethane (20 mL) was added dropwise. The solution was stirred for 36 hours. At this point the reaction was concentrated under reduced pressure to give a crude oil. Column chromatography [silica gel; eluent hexanes-ethyl acetate (3:1)] provided the desired product (0.8 g, 94%) as a clear viscous liquid. 'H(CDCL) 5(ppm): 0.94 (t, 3H); 1.61-1.71 (m, 6H); 2.27 (t, 2H); 2.64 (t, 2H); 4.08 (t, 2H); 7.16- 7.19 (m, 3H); 7.25-7.29 (m, 2H).

Example 2B

Synthesis of 4-(4-Hydroxybutyl)benzoic acid methyl ester 0) 1. ay oO 0 po oH (o]

To aluminum chloride (6.7 g, 0.05 mol) in a dry 250 mL round bottom flask was added anhydrous dichloromethane (100 mL). The flask was cooled in a 0 °C ice bath. Oxalyl chloride (6.4 g, 0.05 mol) was added dropwise to the flask. The mixture was allowed to stir for 5 minutes. A solution of Example 2A (9.8 g, 0.044 mol) in anhydrous dichloromethane (50 mL) was then added dropwise. The mixture was allowed to stir for 4 hours at 0 °C. The reaction mixture was poured into a separatory funnel containing ice and brine. The organic layer was washed with brine and dried over magnesium sulfate. Filtration and concentration under reduced pressure provided 9.1 g of yellow oil. 9.0 g of this oil was suspended in methanol and the pH adjusted to 2 and stirred for 48 hours. The reaction mixture was concentrated under reduced pressure. Column chromatography [silica gel; eluent hexanes-ethyl acetate (2.57:1)] provided the desired product (2.80 g, 31%) as a clear viscous liquid. 'H (CDCl) 8(ppm): 1.56-1.61 (m, 2H); 1.63-1.73 (m, 2H); 2.67 (t, 2H); 3.64 (t, 2H); 3.88 (s, 3H); 7.23 (d, 2H); 7.93 (d, 2H).

Example 2C

Synthesis of 4-[4~tert-Butyldimethylsilanyloxy)butyl]benzoic acid methyl ester oan TB le J

AQ J 0 0) [0]

To Example 2B (1.0 g, 4.8 mmol) was added anhydrous dimethylformamide (10 mL), imidazole (0.5 g, 7.2 mmol) and tert-butyldimethylsilyl chloride (1.08 g, 7.3 mmol). The solution was stirred in a water bath for 2 hours. The reaction mixture was diluted with ethyl acetate, poured into a separatory funnel, washed with water (20 mL, 5x) then washed with a saturated sodium bicarbonate solution (20 mL, 2x).

The organic layer was dried with magnesium sulfate, filtered, and concentrated under reduced pressure to give the desired product (1.17 g, 75 %) which was used without further purification in the next step.

Example 2D

Synthesis of {4-[4~(tert-Butyldimethylsilanyloxy)butyl]phenyl}-methanol oTBS LAH OTBS

To Example 2C (1.17 g, 3.6 mmol) was added anhydrous diethyl ether (14 mL). The solution was cooled to 0 °C with an ice bath. Lithium aluminum hydride (0.28 g, 7.2 mmol) was added to the solution in portions. The mixture was stirred for 1 hour. To the reaction mixture was added distilled water (0.28 mL) and the mixture was stirred for 5 minutes. Next was added an aqueous 15% sodium hydroxide solution and the mixture was stirred for 5 minutes. Lastly distilled water (0.84 mL) was added and the mixture was stirred for 5 minutes. The white solid was removed by filtration. The filtrate was dried with magnesium sulfate, filtered, and concentrated to give 1.23 g of crude product. Column chromatography [silica gel; eluent hexanes-ethyl acetate (4:1)] provided the desired product (1.02 g, 96%) as a clear viscous liquid.

Example 2E

Synthesis of 2-tert-Butyl-5-{4-[4-(tert-butyldimethylsilanyloxy)butyl|benzyloxy}- 4-chloro-2H-pyridazin-3-one

A, i cl ores ChaC0s Fy oTBS

To a dry 25 mL round bottom flask, fitted with a reflux condenser, was added the product of Example 2D (0.41 g, 1.4 mmol), 2-tert-butyl-4,5-dichloro-2H- pyridazin-3-one (0.93 g, 4.2 mmol), cesium carbonate (1.37 g, 4.2 mmol), and anhydrous dimethylformamide (11 mL). The reaction flask was placed in a 68 °C oil bath and the reaction was stirred for 12 hours. The reaction flask was removed from the oil bath and allowed to cool. The mixture was diluted with ethyl acetate, transferred to a separatory funnel and washed with water (25 mL, 5x). The organic layer was dried with magnesium sulfate, filtered, and concentrated under reduced pressure to give 1.3 g of crude product. Column chromatography [silica gel; eluent hexanes-ethyl acetate (9:1)] provided the desired product (594 mg, 89%). 'H(CDCl) (ppm): 0.05 (s, 6H); 0.90 (s, 9H); 1.64 (s, 9H); 2.65 (t, 2H); 3.64 (t, 2H); 5.23 (s, 2H); 7.23 (4, 2H); 7.33 (4, 2H); 7.74 (s, 1H). 13¢ (CDCly) 8(ppm): 18.57, 26.19, 27.75, 28.09, 32.58, 35.61, 63.14, 66.57, 72.14, 118.46, 125.41, 127.44, 129.23, 132.38, 143.72, 154.02, 159.30.

Example 2F

Synthesis of 2-tert-Butyl-4-chloro-5-[4-(4-hydroxy-butyl)-benzyloxy]-2H- pyridazin-3-one %J nr i 9 SL. S— oy

So Ng

TC ome Cn

To the product of Example 2E (594 mg, 1.45 mmol) was added anhydrous tetrahydrofuran (3 mL) and a 1.0 M solution of tert-butylammonium fluoride in tetrahydrofuran (2.9 mL, 2.9 mmol). The solution was stirred for 1 hour then concentrated under reduced pressure. Column chromatography [silica gel; eluent pentane-ethyl acetate (1.8:1)] provided the desired product (410 mg, 77%). 'H (CDCls) 8(ppm): 1.61-1.64 (m, 11H); 1.67-1.74 (m, 2H); 2.68 (t, 2H); 3.68 (t, 2H); 5.23 (s, 2H); 7.23 (d, 2H); 7.33 (4, 2H); 7.74 (s, 1H). BC (CDCl) 8(ppm): 27.43, 27.86, 32.56, 35.35, 62.74, 66.36, 71.88, 118.27, 125.18, 127.27, 128.99, 132.28, 143.17,153.78, 159.07.

Example 2G

Synthesis of Toluene-4-sulfonic acid 4-[4-(1-tert-butyl-5-chloro-6-0x0-1,6- dihydro-pyridazin-4-yloxymethyl)-phenyl]-butyl ester 0 DAP o lane oe oe

Cn AC

To a 5 mL round bottom flask was added the product of Example 2F (200 mg, 0.55 mmol), p-toluenesulfonyl chloride (125 mg, 0.66 mmol), 4-

(dimethylamino)pyridine (80 mg, 0.66 mmol), diisopropylethylamine (85 mg, 0.66 mmol) and anhydrous dichloromethane (2 mL). The resulting solution was stirred for 2 hours. The reaction mixture was diluted with ethyl acetate, transferred to a separatory funnel and washed with a solution of aqueous 0.1 N hydrochloric acid and then washed with brine. The organic layer was dried with magnesium sulfate, filtered, and concentrated under reduced pressure to give 299 mg of crude product.

Column chromatography [silica gel; eluent pentane-ethyl acetate (3:1)] provided the desired product (197 mg, 69%). 111(CDCls) 3(ppm): 1.62-1.70 (m, 13H); 243 (5, 3H); 2.58 (t, 2H); 4.03 (t, 2H); 7.15 (d, 2H); 7.29-7.33 (m, 4H); 7.72 (s, 1H); 7.77 (4, 2H). 13C (CDCl) 8(ppm): 21.63, 26.98, 27.86, 28.34, 34.80, 66.37, 70.23, 71,81, 118.25, 125.12, 127.32, 127.87, 128.93, 129.82, 132.48, 133.15, 142.40, 144.72, 153.75, 159.05.

Example 2H

Synthesis of 2-tert-butyl-4-chloro-5-(4-(4-fluorobutyl)benzyloxy 3(2H) pyridazinone : 0}

Ky Lo KF-K222 Fy

Um, F

The product of Example 2G (57 mg, 0.10 mmol) was dissolved in 1 mL acetonitrile and to this was added a mixture of KF-K222 (1:1; 0.164 mmol) dissolved in 1 mL acetonitrile. The entire mixture was then immersed in an oil bath at 90 °C and heated at reflux for 15 minutes at which point the reaction was shown to be complete by TLC. The volatile components were removed in vacuo and the crude oil was purified by flash silica gel chromatography (hexanes-ethyl acetate 4:1) to provide 28 mg of the desired product as a oil which solidified upon standing. 'H (CDCl3) (ppm): 1.6 (s, 9H), 1.7 (m, 4H), 2.6 (t, 2H), 4.44 (d of t, 2H, J=47.4 & 6

Hz), 5.2 (s, 2H), 7.2 (4, 2H, J = 8.4 Hz), 7.3 (4, 2H, J = 8.4 Hz), 7.71 (s, 1H). BC (CDCls) 8(ppm): 26.8 (Jer = 4.65 Hz), 27.8, 29.8(Jcr= 19.8 Hz), 35.1, 66.3,71.8, 83.8 (Jer = 163.8 Hz), 118.2, 125.1, 127.2, 128.9, 132.3, 142.8, 153, 159. 9E(CDCl , CFCly as internal standard) 8(ppm): - 218.6 (toft, J = -27.6, -50.4)

Example 3A

Synthesis of (2)-1-tert-butyldimethylsilyloxy-2-hydroxybutane

TBSCI, imidazole

ER on

A 50mL round bottom flask was charged with (£)-1 ,2-butanediol (1g, 11.09 mmol) and to it was added dimethylformamide (8mL) followed by tert- butyldimethylsilyl chloride (2.5g, 16.64 mmol) and imidazole (1 .88g, 27.7 mmol).

The reaction mixture was stirred for 10 hours after which it was diluted with dichloromethane and poured into a separatory funnel and washed with water (80 mL) and brine and dried over magnesium sulfate. After filtration and concentration the _ crude oil was purified by silica gel flash chromatography (hexanes:ethylacetate) to obtain 1gm of pure desired product in 45% yield. '{ (CDCl) § (ppm): 3.6 (m, 1H). 3.5 (m, 1H), 3.4 (m, 1H), 2.4 (s, 1H), 1.44 (m, 2H), 0.99 (¢, 3H), 0.9 (s, 9H), 0.06 (s, 6H).

Example 3B

Synthesis of (£)-4-(1-tertbutyldimethylsityloxy but-2-oxy) methylbenzoate

MeO. 0 MeO._.0

PPhs, DIAD & I &

OH oy ome 4-Hydroxymethylbenzoate (1.1g, 7.34 mmol), the product of Example 3A (0.75g, 3.67 mmol) and triphenylphosphine (1.972 g, 7.34 mmol) were added to a round bottom flask and 8 mL tetrahydrofuran was added. The flask was cooled in an ice bath to 0 °C after which diisopropylazodicarboxylate (1.485g, 7.34mmol) was added via syringe. The reaction mixture was stirred for 2 hours after which the reaction was deemed complete by thin layer chromatography. All the solvent was removed under reduced pressure and the crude oil directly subjected to purification by silica gel flash chromatography (hexanes : diethyl ether) to obtain 1.0 gm (83%) of the desired compound as a thick oil. 'H (CDCls) 8 (ppm): 7.9 (4, 2H), 6.9 (d, 2H), 4.3 (p, 1H,J =5.4 Hz), 3.9 (s, 3H), 3.7 (2H), 1.78 (m, 1H), 1.7 (m, 1H), 0.9 (t,3H,J = 7.8 Hz), 0.89 (s, 9H), 0.05 (s, 3H), 0.01 (s, 3H). 13¢ (CDCls) 8 (ppm): 166.8, 162.8, 131.5, 122.3, 115.2, 80, 64.5, 51.7, 25.8, 24.1,18.2,9.5,-5.3.

Example 3C

Synthesis of (£)4-(1-tertbutyldimethylsilyloxy but-2-oxy) benzylalcohol

MeO._.0 HO : S LAH, Et,0 " yo ye

To a solution of the product of Example 3B (1g, 2.95 mmol) in ether (15mL) was added lithium aluminum hydride (0.336g, 8.8 mmol) and the mixture was stirred under nitrogen for 1.5 hours. The reaction was complete as shown by TLC by this time and was quenched by addition of 0.336 mL water, 0.336 mL of 15% NaOH solution and 1.00 mL water in succession. The resulting mixture was stirred for an additional 20 minutes after which the white precipitate formed was filtered and washed with ether. The filtrate was then dried over magnesium sulfate. Filtration and removal of the solvent gave 0.50g (54%) of the desired product as a white solid.

IH (CDCl) & (ppm): 72 (d, 2H), 6.9 (d, 2H), 4.3 (p, 1H), 3.77(d of d, 1H), 3.66 (d of d, 1H), 1.77-1.72 (m, 1H), 1.68-1.61 (m, 1H), 1.5(t, 1H, J = 5.4 Hz), 0.9 (t, 3H, J = 7.8 Hz), 0.89 (s, 9H), 0.04 (s, 3H), 0.01 (5, 3H). BBC (CDCl) 8 (ppm): 158.5, 133, 128.4, 116.1, 80.1, 65, 64.5, 25.8, 24.1, 18.2,9.5,-53

Example 3D

Synthesis of (+)-2-tert-butyl 4-chloro 5-(4(1-tertbutyldimethylsilyloxy but-2- oxy) benzyl)oxy 3(2H)-pyridazinone lo} i fo) 206 * > ee 206 oTBS ye : (+)-2-Tert-butyl-4-chloro-5-hydroxy-3 (2H)-pyridazinone (0.48g, 2.417 mmol) was charged to a 100 mL round bottom flask and tetrahydrofuran (40ml) was added. After the solution turned clear, Example 3C (0.5g, 1.611 mmol) and triphenylphosphine (0.633g, 2.417 mmol) were added to the flask and the flask was cooled to 0°C. Diisopropyl azodicarboxylate (0.488g, 2.417 mmol, 0.468 mL) was then added via a syringe and the reaction was stirred for two hours after which time it was shown to be complete by TLC. The contents of the flask were then concentrated in vacuo and the crude oil obtained was purified by flash chromatography using silica gel (hexanes:ethyl acetate) to obtain 0.33 g of the desired compound as an oil. 'H (CDCl) & (ppm): 7.72 Gs, 1H), 7.2 (4, 2H), 6.9 (d, 2H), 5.2 (s, 2H), 4.2 (p, 1H), 3.75 « of 4, 1H), 3.68 (d of 4, 1H), 1.75 (m, 2H), 1.65 (m, 1H), 1.6 (s, 9H), 0.99 (t, 3H), 0.85 (s, 9H), 0.04 (s, 3H), 0.02 (s, 3H). 13C (CDCls) 8 (ppm): 159.6, 159.3, 154, 129, 126.9, 125, 118.5, 116.5, 80.3, 72.1, 66.5, 64.8, 28.1, 26, 24.4, 18.4, 9.6,-5.3

Example 3E

Synthesis of (£)-2-tert-butyl-4-chloro-5-(4-(1-hydroxy-but-2-oxy)benzyloxy- 3(2H)-pyridazinone o 0} 206 ors TBAFInTHF 206

N “CL C TL Ng CL C 0 0

To the product of Example 3D (0.3 g, 0.6 mmol) ina 10 mL round bottom flask was added tetrahydrofuran (2 mL). Upon solution, tetrabutylammonium fluoride (1.8 mmol, 1.8 mL, IM solution in THF) was added and the reaction mixture was stirred for 90 minutes. The contents were then concentrated under reduced pressure and the crude mixture purified by flash chromatography using silica gel (hexanes:ethyl acetate) to obtain 185 mg (80%) of pure desired product. 'H (CDCl) 3 (ppm): 7.74 (s, 1H), 7.3 (4, 2H), 6.9 (d, 2H), 5.2 (s, 2H), 4.3 (m, 1H), 3.81-3.77 (two br s, 2H), 1.84 (br t, 1H), 1.77-1.69 (m, 2H), 1.64 (5, 9H), 0.98 (t, 3H); "°C (CDCl) 8 (ppm): 159.2, 158.9, 153.9, 129.2, 127.5, 125.4, 116.6, 80.4, 71.9, 66.5, 64.2,28,23.5,9.7.

Example 3F

Synthesis of (1)-2-tert-butyl 4-chloro 5-(4-(1-tosyloxy-but-2-oxy) benzyl)oxy 3(2H)-pyridazinone lo

AA TsCl, DMAP S66 ey oH “oom DEA aaa ol OTs

PS oA

Into a 10 mL round bottom flask was added the product of Example 3E (0.05g, 0.13 mmol) followed by dichloromethane (2 mL). Toluenesulfonyl chloride

(0.075g, 0.39 mmol), 4-N,N-dimethylaminopyridine (0.048g, 0.39 mmol) and diisopropylethylamine (0.05g, 0.39 mmol, 68.711) were then added in succession to the reaction mixture and this was stirred for 35 minutes. Water was then added to the mixture and the solution poured into 2 separatory funnel and the layers separated. :

The organic layer was washed with water and brine and dried over magnesium sulfate. The crude oil obtained after filtration and concentration was purified by silica gel flash chromatography (hexanes:ethyl acetate) to obtain 54 mg (77%) of the desired compound as a thick colorless oil. 'H (CDCls) 8 (ppm): 7.74 (3H, two singlets), 7.3 (m, 4H), 6.8 (d, 2H), 5.2 (s, 2H), 4.38 (p, 1H), 4.15 (m, 2H), 2.44 (s, 3H), 1.72 (m, 2H), 1.6 (5, 9H), 0.95 (t, 3H); BC (CDCl) 8 (ppm): 159.2, 158.5, 153.9, 145.1, 133, 130, 129, 128.1, 127.2, 125.4, 118.5, 116.5, 71.9, 70.2, 66.6, 28.1, 24.2,21.8,9.4.

Example 3G

Synthesis of (2)-2-tert-butyl-4-chloro 5-(4-(1-flnoro-but-2-oxy)benzyloxy-32H)- pyridazinone 0 0}

Aye KFK222 Aye s “CL KS AcN, 90C S “CL KS 0 0

The product of Example 3F (28mg, 52.4 pmol) was dissolved in 0.5 mL acetonitrile in a 5 mL flask and to this was added a solution of potassium fluoride (4.5 mg, 78.6 pmol) and Kryptofix 222 (29.6 mg, 7 8.6 pmol) in 0.5 mL acetonitrile.

The above solution was then immersed in a oil bath preheated to 90 °C. The reaction was allowed to stir for 90 minutes after which all the volatiles were removed under reduced pressure and the crude mixture purified by preparative thin layer chromatography to obtain 13 mg (65%) of pure desired compound. 'H (CDCl) § (ppm): 7.72 (s, 1H), 7.3 (d, 2H), 6.9 (4, 2H), 5.23 (s, 2H), 4.57-4.59 (m, 2H), 4.4 (m, 4H), 1.74 (m, 2H), 1.6 (s, 9H), 1.0 (+, 3H). 13¢ (CDCl) 8 (ppm): 159, 158.7, 153.7, 129, 127.5, 125.2, 118.3, 116.4, 83.85 (d, yep =172.2),78,71.1, 66.3,27.8,23.2, 9.48. °F (CDCl, CFCl, as internal standard) 8 (ppm): -228 (d of t, J =-19, -60 Hz )

Example 4A

Synthesis of 4-(3-hydroxypropoxy)-benzoic acid methyl ester

Q KoCOs 0 _~_POH jond on DME, Jes

HO o

To a 250 mL flask was added 3.bromo-1-propanol (4.17 g, 0.03 mol), anhydrous dimethylformamide (40 mL), methyl-4-hydroxybenzoate (3.0 g, 0.02 mol) and potassium carbonate (4.15 g, 0.03 mol). The flask was placed in a 50 °C oil bath and stirred for 12 hours. After cooling the reaction was diluted with ethyl acetate, transferred to separatory funnel, washed with aqueous 0.1 N hydrochloric acid, water then brine. The organic layer was dried with magnesium sulfate, filtered, and concentrated under reduced pressure to give 5.14 g of crude oil. Column chromatography [silica gel; eluent hexanes-ethyl acetate (1.68:1)] provided the desired product (1.25 g, 30 %) as a white powder. 'H (CDCl3) 8(ppm): 2.04-2.08 (m, 2H); 3.86-3.88 (x, SH); 4.17 (¢, 2H); 6.91 (4, 2H); 7.98 (d, 2H); "*C (CDCls) 3(ppm): 31.89, 51.81, 59.88, 65.50, 114.06, 122.67, 131.57, 162.60, 166.84.

Example 4B

Synthesis of 4-[3-(tert-Butyldimethylsilanyloxy)propoxy]benzoic acid methyl ester

OOH sole 0 _~_0OTBS oA or - 2 AJ lo} o]

To a 50 mL flask was added Example 4A (300 mg, 1.4 mmol), anhydrous dimethylformamide (4 mL), tert-butyldimethylsilyl chloride (317 mg, 2.1 mmol), and imidazole (146 mg, 2.1 mmol). The resulting solution was stirred for 2 hours. At this point the reaction was diluted with ethyl acetate and transferred to a separatory funnel. The organic phase was washed with aqueous 0.1 N hydrochloric acid(2x), water(2x), then brine. The organic layer was then dried over magnesium sulfate, filtered, and concentrated. Column chromatography [silica gel; eluent hexanes-ethyl acetate (9.5:1)] provided the desired product (413 mg, 91 %). 'H (CDCL) (ppm): 0.03 (s, 6H); 0.87 (s, 9H); 1.97-2.01 (m, 2H); 3.79 (t, 2H); 3.87 (s, 3H); 4.11 (t, 2H); 6.90 (d, 2H); 7.97 (4, 2H); "°C (CDCl) 8(ppm): 18.30, 25.89, 32.3, 51.78, 59.27,

64.67, 114.08, 122.43, 131.56, 162.90, 166.90

Example 4C

Synthesis of (4-[3tert-Butyldimethylsilanyloxy)propoxy]phenyl} methanol

O~_PTBS LAH oo

PC pau SEE (0)

Example 4B (396 mg, 1.22 mmol) was added to a dry 50 mL flask along with anhydrous diethyl ether (10 mL). The flask was lowered into an ice bath. Lithium aluminum hydride (93 mg, 2.44 mmol) was added in portions to the reaction flask.

The mixture was allowed to stir in the bath for 2 hours. The reaction was quenched with water (0.093 mL), aqueous 15 9, sodium hydroxide (0.093 mL) then water (0.279 mL). The white solid was filtered off and the filtrate was dried over magnesium sulfate, filtered, and concentrated to give the desired product (291 mg, 80 %)- 'H(CDCl;) (ppm): 0.04 (s, 6H); 0.88 (s, 9H); 1.95-1.99 (m, 2H); 3.79 (4, 2H); 4.05 (t, 2H); 4.60 (s, 2H); 6.88-6.89 (m, 2H); 7.25-7.27 (m, 2H); (CDCl) 8(ppm): 18.30, 25.91, 32.41, 59.50, 64.57, 65.10, 114.59, 128.60, 132.97, 158.75.

Example 4D :

Synthesis of 2-tert-butyl-4-chloro-5-{4-[3-(tert- butyldimethylsilanyloxy)propoxy]benzyloxy}-2H-pyridazin-3-one

PPh

UE SR Re wow IJ XX, “oy, 0 ~"oTBS

To a dry 25 mL flask was added Example 4C (211 mg, 0.71 mmol) and anhydrous tetrahydrofuran (3 mL). The flask was cooled in an ice bath. To the flask was added triphenylphosphine (187 mg, 0.71 mmol) and 2-tert-butyl-4-chloro-3- hydroxy-2H-pyridazin-3-one (142 mg, 0.71 mmol). Lastly, diisopropyl azodicarboxylate (144 mg, 0.71 mmol) was added. The reaction mixture was allowed to stir in the ice bath for 1 hour. At this point the mixture was diluted with diethyl ether and transferred to a separatory funnel. The organic solution was washed with water and then brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Column chromatography [silica gel; eluent hexanes-ethyl acetate (9:1)] provided the desired product (106 mg, 31 %). 'H (CDCl3) 8(ppm): 0.03 (s, 6H); 0.87 (s, 9H); 1.62 (s, 9H); 1.95-1.99 (m, 2H); 3.79 (t, 2H); 4.06 (t, 2H); 5.23

(s, 2H); 6.91-6.92 (m, 2H); 7.30-7.31 (m, 2H); 7.72 (s, 1H); °C (CDCl) (ppm): 18.29, 25.90, 27.87, 32.34, 59.41, 64.63, 66.30, 71.89, 114.90, 118.34, 125.34, 126.68, 128.92, 153.79, 159.07, 159.55

Example 4E

Synthesis of 2-tert-butyl-4-chloro-5-[4-(3-hydroxypropoxy)-benzyloxy}-2B- pyridazin-3-one 0 TBAF o

Fy = Oy

Ny CL Na CL 0 ~"oTBS 00H

To a dry 10 mL flask was added Example 4D (100 mg, 0.21 mmol) along with anhydrous tetrahydrofuran (2 mL). To the flask was added a solution of 1.0M tetrabutylammonium fluoride in tetrahydrofuran (0.42 mL, 0.42 mmol). The solution was stirred for 2 hours. At this point the reaction was concentrated under reduced pressure. Preparatory thin layer chromatography [silica gel; eluent hexanes-ethyl acetate (1:1)] provided the desired product (57.8 mg, 76 %). 'H(CDCl) 8(ppm): 1.62 (s, 9H); 2.02-2.06 (m, 2H); 3.86 (t, 2H); 4.13 (t, 2H); 5.30 Gs, 2H); 6.92-6.93 (m, 2H); 7.31-7.32 (m, 2H); 7.71 (s, 1H); °C (CDCL3) 8(ppm): 27.87, 31.97, 60.24, 65.67, 66.34, 71.81, 114.91, 118.37, 125.31, 127.06, 128.98, 153.76, 159.07, 159.27.

Example 4F

Synthesis of toluene-4-sulfonic acid 3.[4-(1-tert-butyl-5-chloro-6-0x0-1,6- dihydro-pyridazin-4-yloxymethyl)phenoxylpropyl ester

TsCl 0 rl °

Fy ye DCM Fa re 0 "0H TC on

To a dry 5 mL flask was added Example 4E ( 40 mg, 0.11 mmol), 4-methyl- benzenesulfonyl chloride (31 mg, 0.16 mmol), 4-(dimethylamino)pyridine (20 mg, 0.16 mmol), diisopropylethylamine (16.6 mg, 0.16 mmol) and anhydrous dichloromethane (0.6 mL). The resulting solution was stirred for 1 hour. The reaction mixture was concentrated under reduced pressure. Preparatory thin layer chromatography [silica gel; eluent pentane-ethyl acetate (3:2)] provided the desired product (18.6 mg, 33%). 'H (CDCl) 8(ppm): 1.62 (s, 9H); 2.09-2.13 (m, 2H); 2.37

(s, 3H); 3.95 (t, 2H); 4.23 (4, 2H); 5.22 (s, 2H); 6.78 (d, 2H); 7.23 (d, 2H); 7.29 (d, 2H); 7.73-7.75 (m, 3H). °C (CDCl) (ppm): 21.60, 27.85, 28.81, 63.15, 66.35, 66.87, 71.75, 114.76, 118.27, 125.18, 127.11, 127.83, 128.94, 129.80, 132.79, 144.80, 163.72, 158.90, 159.03.

Example 4G

Synthesis of 2-tert-butyl-4-chloro-5-[4-(3-flucropropoxy)benzyloxy]-2H- pyridazin-3-one 1% ke22 fo}

Fy ACN Fo Se

Naor ® Ny 0 ""0Ts ASW.

To a scintillation vial containing a suspension of Example 4F (4.5 mg, 8.64 x 10? mmol) in anhydrous acetonitrile (0.25 mL) was added a solution of potassium fluoride (1.6 mg, 4.07 x 10° mmol) and kryptofix (15.0 mg, 4.07 x 102 mmol) in anhydrous acetonitrile (0.25 mL). The vial was capped and lowered into a 90 °C oil bath. The reaction was allowed to stir for 40 minutes. The reaction was cooled and concentrated under reduced pressure. Preparatory thin layer chromatography [silica gel; eluent pentane-ethyl acetate (3:2)] provided the desired product (0.8 mg, 25 %). 'H(CDCl;) 3(ppm): 1.62 (s, 9H); 2.14-2.20 (m, 2H); 4.09-4.11 (m, 2H); 4.60 (t, 1H); 4.68 (t, 1H); 5.24 (s, 2H); 6.92 (d, 2H); 7.32 (4, 2H); 7.72. (5, 1H); ’F(CDCl;, CFCl as internal standard) 5(ppm): -222.66 (t of t, J = 28.2, -50.4)

Example SA

Synthesis of 4-(2-hydroxyethoxymethyl)benzoic acid methyl ester

CO,Me CO,Me

Co

C BF Et,0 Q

OH a

To a two-neck round bottom flask, which was equipped with a Dewar condenser, a solution of 4-hydroxymethylbenzoic acid methyl ester (2.50 g, 0.015 mol) in anhydrous dichloromethane (30 mL) was cooled to -10 °C in a salt/ice bath.

Ethylene oxide (1.10 mL) was added to the cooled stirring solution dropwise followed by the addition of boron trifluoride etherate (0.51 ml). The reaction mixture was stirred for 45 minutes and then warmed to room temperature for 30 minutes to boil off any excess of ethylene oxide in the reaction mixture. The reaction mixture was then diluted with brine. The aqueous layer was extracted with dichloromethane (3 times). All of the organic layers were combined, dried over Na;SOs, filtered, and concentrated to provide an oil. The crude material was purified using silica gel chromatography (4:1 pentane:ethyl acetate) to provide the desired product (537 mg, 2.56 mmol) in 17% yield. 'H (CDC1:8.36, 600 MHz): 8 (2H, d, J=8.4 Hz), 7.41 (2H, d, J=8.5 Hz), 4.62 (3H, 5), 3.92 (2H, 5), 3.78 (m, 2H), 3.63 (2H, m); Bc (CDC1L:167.1, 143.5, 130.0, 129.8, 127.5, 72.9, 72.0,, 150 MHz): 62.1, 52.3.

Example 5B

Synthesis of 4-[2(ter+-butyldimethylsilanyloxy)ethoxymethyl]benzoic acid methyl ester

CO,Me CO Me ® TBDMS-Ci, . imidazole, DMF o A~_oH ~_OTBDMS

To a solution of the product of Example 5A (544.5 mg, 2.59 mmol) in anhydrous DMF (26 mL) was added imidazole (264 mg, 3.89 mmol) and TBDMS-Cl (586 mg, 3.89 mmol). The reaction mixture stirred at room temperature overnight and was quenched with water. The aqueous layer was extracted with ethyl acetate (3x). All combined organic layers were dried over Na,;SOs, filtered, and concentrated. The crude material was purified using silica gel chromatography (4:1 pentane:ethyl acetate) to provide the desired product (67 7.5 mg, 2.19 mmol) in 84% yield. 'H (CDC1:8.01, 600 MHz): 3 (2H, d, J=8.3 Hz), 7.42 (2H, d, J=8.4 Hz), 4.63 (2H, s), 3.91 (2H, 5), 3.82 (2H, t, J=5.0), 3.58 (2H, t, J=5.1 Hz), 0.91 (9H, s), 0.07 (6H, s); °C (CDCl;166.5, 143.5, 129.2, 128.8, 126.5, 72.1, 71.6,, 150 MHz): § 62.3, 51.5,25.4,17.9,-5.8.

Example SC

Synthesis of (4-[2-(tert-butyldimethylsilanyloxy)ethoxymethyl]phenyljmethanol

CO, Me H

Q mi S o ~_-OTeDMS o ~_-OTBDMS

To a solution of the product of Example SB (670 mg, 2.18 mmol) dissolved in anhydrous THF (22 mL) was added a solution of LAH (1.0 M solution in THF, 2.18 mL, 2.18 mmol) dropwise. After completion of addition the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with water.

The aqueous layer was extracted with ethyl acetate (3x). All combined organic layers were dried over Na;SOs, filtered, and concentrated to provide an oil (587 mg, 1.98 mmol), which was used in the next step without any further purification (91% yield). 1H (CDCl; 7.34 (4H, 5), 4.68 (2H, 5), 4.57 (2H, 5), 3.80, 600 MHz): 8 (2H, t, J=52

Hz), 3.56 (2H, t, J=5.3 Hz), 1.69 (1H, br 8), 0.90 (9H, 5), 0.07 (6H, s); *C (CDCl; 140.4, 138.3, 128.0, 127.2, 73.2, 71.9, 65.4,, 150 MHz): § 63.0, 26.2, 18.6, -5.0.,

Example 5D

Synthesis of 2-tert-butyl-5-{4-[2-(tert- butyldimethylsilanyloxy)ethoxymethyl]benzyloxy}-4-chloro-2H-pyridazin-3-one

Q 0

FY ee ddl NY OTBDMS

N~ OH o Nx o - o~OTBDMS AY

To solution of the product of Example 5C (437 mg, 1.48 mmol) and 2-tert- butyl-4-chloro-5-hydroxy-2H-pyridazin-3-one (250 mg, 1.23 mmol) dissolved in anhydrous THF (12 mL) was added solid PPh; (485 mg, 1.85 mmol) and diisopropyl azodicarboxylate (DIAD, 0.358 mL, 1.85 mmol). After completion of addition the reaction mixture continued to stir at room temperature. After 20 hours, the reaction mixture was diluted with water. The aqueous layer was separated and extracted with ethyl acetate (3x). All combined organic layers were dried over NaySOs, filtered, and concentrated to provide an oil. The crude material was purified using silica gel chromatography (4:1 pentane: ethyl acetate) to provide the desired product 528 mg, 1.10 mmol) in 89% yield. 'H (CDCl; 7.70 (1H, 5), 7.38 (4H, m), 5.30 CH, 5), 4.58, 600 MHz): 5 (2H, 5), 3.80 2H, t, J= 5.4 Hz), 3.57 2H,t,J=5.4 Hz), 1.63 (9H, br s), 0.90 (9H, s), 0.07 (6H, s); BC (CDC);159.0, 153.7, 138.8, 134.4, 128.3,127.3,,150

MHz): § 125.1, 118.5,72.8, 71.7, 71.6, 66.4, 61.9, 29.7, 27.9, 25.6, -5.1.; HRMS calcd for CasHa7CIN2O4Si: 481.228389, found 481.2282.

Example SE

Synthesis of 2- tert-butyl-4-chloro-5-[4-(2-hydroxyethoxymethyhbenzyloxy]-2H- pyridazin-3-one 0 0

TOU

To a solution of the product of Example 5D (528 mg, 1.09 mmol) dissolved in anhydrous THF (11 mL) was added a solution of TBAF (1.0 M solution in THF, 1.65 mL, 1.65 mmol) dropwise. After completion of addition the reaction was stirred at room temperature for 1 hour and then quenched with water. The aqueous layer was separated and extracted with ethyl acetate (3x). All combined organic layers were dried over Na,SO,, filtered, and concentrated to provide an oil. The crude material was purified using silica gel chromatography (4:1 hexanes: ethyl acetate) to provide the desired product (311 mg, 0.850 mmol) in 78% yield. 'H (CDCl, 600 MHz): 1) 7.70 (1H, s), 7.38 (4H, m), 5.30 (2H, 5), 4.56 (2H, s), 3.76 (2H, t, J=4.9 Hz), 3.60 (2H, t, J=4.8 Hz), 2.00 (1H, brs), 1.61 (9H, brs); 13C (CDCL3159.0, 153.6,, 150

MHz): 5 138.8, 134.4, 128.2, 127.2, 125.1, 118.3, 72.8, 71.6, 71.6, 66.4, 61.9, 27.8;

HRMS calcd for C,sH23CIN;O4: 367.141911, found 367.1419.

Example SF

Synthesis of toluene-4-sulfonic acid 2-[4-(1-tert-butyl-5-chloro-6-oxe-1,6- dihydro-pyridazin-4-yloxymethyl)-benzyloxy}-ethyl ester [lo] lg J we gf

LN rut LN

To a solution of the product of Example SE (200 mg, 0.546 mmol) dissolved in anhydrous dichloromethane (5.50 mL) was added TsCl (125 mg, 0.656 mmol),

DMAP (100 mg, 0.819 mmol) and triethylamine (0.091 mL, 0.656 mmol). The reaction mixture continued stirring at room temperature. After 22 hours the reaction mixture was diluted with water. The aqueous layer was separated and extracted with ethyl acetate (3x). All combined organic layers were dried over Na,SO4, filtered, and concentrated to provide an oil. The crude material was purified using silica gel chromatography (3:2 pentane:ethyl acetate) to provide the desired product (232 mg, 0.447 mmol) in 82% yield. 'H (CDC1;7.79, 600 MHz): § (2H, d, J=8.3 Hz), 7.71 (1H, s), 7.38 (2H, 4, J=8.2 Hz), 7 32 (4H, m), 5.30 (2H, 5), 4.50 (2H, 5), 4.21 (2H, m), 3.69 (2H, m), 2.43 (3H, 5), 1.63 (9H, br s); 13C (CDCl; 159.0, 153.7, 144.8, 138.8,, 150 MHz): 5 134.4, 133.1, 129.8, 128.1, 128.0, 127.2, 125.1, 118.4, 72.8, 71.7, 69.2, 67.8, 66.4, 27.9, 21.6; HRMS calcd for CasHaeCIN;O6: 521.150762, found 521.1503.

Example 5G

Synthesis of 2-tert-butyl-4-chloro-5-[4-(2-fluoro-ethoxymethyl)-benzyloy]-2H- pyridazin-3-one

[0] [o]

To a solution of the product of Example 5F (50 mg, 0.096 mmol) in anhydrous acetonitrile (1.0 mL) was added KF (11.2 mg, 0.192 mmol) and Kryptofix (72.4 mg, 0.192 mmol). After completion of addition the reaction mixture was heated to 90 °C. After 10 minutes, the reaction mixture was cooled down to room temperature and diluted with water. The aqueous layer was separated and extracted with ethyl acetate (3x). All combined organic layers were dried over Na,;SOs, filtered, and concentrated to provide an oil. The crude material was purified using silica gel chromatography (4:1 pentane: ethyl acetate) to provide the desired product (28 mg, 0.076 mmol) in 79% yield. 'H (DMSO-d, 600 MHz): 8 8.22 (1H, s), 7.45 (2H, d, J=8.20 Hz), 7.39 (2H, d, J=8.24 Hz), 5.42 (2H, s), 4.60 (1H, m), 4.54 (2H, s), 4.52 (1H, m), 3.71 (1H, m), 3.66 (1H, m), 1.57 (9H, s); 13157.8,153.8,138.6,C (DMSO0-d6, 150 MHz): § 134.6, 127.8, 127.7, 126.2, 115.6, 83.5 (82.4), 71.6, 71.2, 69.1 (69.0), 65.3, 27.4; °F (DMSO-d-221.74 (IF, m)., 564 MHz): & HRMS calcd for C1sH22CIFN;03: 369.137575, found 369.1377.

Example 6A

Synthesis of 1-(4-hydroxymethylphenoxy)propan-2-one

OH

HO. 8 3

Acetone

BR

To a stirred solution of 4-hydroxybenzyl alcohol (1.0 g, 8.06 mmol) in acetone (80 mL) was added potassium carbonate (1.34 g, 9.68 mmol) and chloroacetone (0.771 mL, 9.68 mmol). After completion of addition the reaction mixture was heated to reflux. After 20 hours the reaction mixture was cooled down to room temperature and the solvent was removed. Water and ethyl acetate were added to the crude material. The aqueous layer was separated and extracted with ethyl acetate (3x, 100 mL). All combined organic layers were dried over Na,SO4, filtered, and concentrated to provide an oil. The crude material was purified using silica gel chromatography (gradient from 4:1 to 1:1 pentane:ethyl acetate) to provide the desired product (0.981 g, 5.45 mmol) in 98% yield. 'H (CDCls;, 600 MHz): 8 7.30 (2H, d, J=8.7 Hz), 6.87 (2H, d, J=8.7 Hz), 4.63 (2H, d, J=5.7 Hz), 4.54 (2H, s), 2.27 (3H, 5), 1.66 (1H, t, J=5.8 Hz); °C (CDCl, 150 MHz): 5205.7, 157.3, 134.3, 128.8, 114.6, 73.1, 64.8, 26.6.

Example 6B

Synthesis of 1-(4-hydroxymethyl-phenoxy)-propan-2-ol:

OH H

S NaBH,, MeOH S

BY

: BE

To a solution of 1-(4-hydroxymethylphenoxy)-propan-2-one (1.26 g,6.99 mmol) dissolved in methanol (60 mL) was added solid NaBH, (0.32 g, 8.39 mmol).

Claims (24)

PCT/US2005/004687 WHAT IS CLAIMED IS:

1. A contrast agent comprising an imaging moiety and a compound selected from deguelin, pyridaben, pyridimifen, tebufenpyrad, fenazaquin a deguelin analog, a pyridaben analog, a pyridimifen analog, a tebufenpyrad analog, and an fenazaquin analog.

2. The contrast agent of claim 1 wherein the imaging moiety is a radioisotope for nuclear medicine imaging, a paramagnetic species for use in MRI imaging, an echogenic entity for use in ultrasound imaging, a fluorescent entity for use in fluorescence imaging, or a light-active entity for use in optical imaging.

3. The contrast agent of claim 2 wherein the paramagnetic species for use in MRI imaging is Gd**, Fe**, In’*, or Mn*".

4. The contrast agent of claim 2 wherein the echogenic entity for use in ultrasound imaging is a fluorocarbon encapsulated surfactant microsphere.

S. The contrast agent of claim 2 wherein the radioisotope for nuclear medicine imaging is tHe, BN, IE, 123 1251, ome, ST, "In, 2c, Cu, Ga, or ¥Ga.

6. The contrast agent of claim 1 wherein the imaging moiety is '*F.

7. The contrast agent of claim 1 wherein the imaging moiety 1s a

8. The contrast agent of claim 1 of formula (I) AMENDED SHEET

PCT/US2005/004687 ® wR RS ] REN A R3 R* "a E A A A : AJC E R? SGN: f R14

Cc . A (0, wherein each A is independently selected from O, CHR, S, and NR'; B is selected from hydrogen, C,-Cg¢ alkyl optionally substituted with an imaging moiety, and an imaging moiety; C is selected from hydrogen, C;-C¢ alkyl optionally substituted with an imaging moiety, an imaging moiety, and a bond to B; D is selected from hydrogen, C;-Cg alkyl optionally substituted with an imaging moiety, and an imaging moiety; E is selected from hydrogen, C,-C¢ alkyl optionally substituted with an imaging moiety, and an imaging moiety; or E and D, together with the carbon atom to which they are attached, form a double bond; or E and D, together with the carbon atom to which they are attached, form a cyclopropyl ring; EN 1s a single or a double bond: R', R:, R’, RY R’, R', R'3 , and RY", are each independently selected from hydrogen, C;-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; R® and R® are each independently selected from hydrogen, C,-C alkyl optionally substituted with an imaging moiety, halo, hydroxy, and an imaging moiety; when present, R” and R® are independently selected from hydrogen, C,-Cs alkyl optionally substituted with an imaging moiety, halo, hydroxy, and an imaging moiety; or AMENDED SHEET

PCT/US2005/004687 @® R® and R’ together form an oxo group; or R® and R® together form an oxo group; or R’ is O and R® is a bond to R; provided that when 2 is a double bond, R’ and R® are absent; R'! is hydrogen or hydroxy; R!'? is selected from hydrogen, C,;-Cg alkyl optionally substituted with an imaging moiety, and an imaging moiety; or R'! and R'? together form an oxo group or =CHR; with the proviso that at least one imaging moiety is present in formula (I).

9. The contrast agent of claim 8, wherein Ais O; B and C are each independently CH; or CH-"*F; D and E are each independently CH; or CH '®F; R3, R®, R’, and R'? are each independently hydrogen or °F; and R'! and R'? together form an oxo group.

10. The contrast agent of claim | wherein the contrast agent is selected from 18 18 ! H : 0 [o] H , Fe) Lo) o a Ee) 0 0 IX Po oN NT i Se ; = d i I NYY H = i ! oo o ad oP 0 Lo , o. ’ PT" ,

~~. ~~ 18g EEL CN or On 0 Mo o 0 ~~ we IN ] 0” ST 1 RS 3 INC, E i Nh I To : pz 0” NF 0 0 o ] , lo , ° ; os ~ | 0 = = (0 oY IN : H - NF

[0] ~~ [o} I oY ~ ’ and Ong : 18g AMENDED SHEET

PCT/US2005/004687

11. The contrast agent of claim 1 of formula (II), R¥® R R23 G R24 ~N J n K R% / ~ R25 / L (ID), wherein 34 RF 1 R2 4 Q N N Ny N~“ a R33 3 b | ~

AN . <% R28 ~ R? R¥ RE Gis " or rR , wherein misOorl; CH _b. == and —— each independently represent a single or a double bond;

R*. RY R*' R*. R*, and R* are independently selected from hydrogen, C,-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; when present, R™ is selected from hydrogen and C,-C, alkyl optionally b a substituted with an imaging moiety, provided that when === is a double bond, R% is absent; when present, R% is C,-Cq alkyl optionally substituted with an imaging moiety, provided that when 2 is a double bond, R* is absent; R36 RY R33 Pis 3 , wherein R%, R¥, RY, R*®, and R*® are AMENDED SHEET

PCT/US2005/004687 ( independently selected from hydrogen, Ci-Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; when present, P’ is hydrogen; or P and P’ together form an oxo group; provided that when 2 is a double bond, P’ is absent; Q is halo or haloalkyl; J is selected from N(R), S, 0, C(=0), C(=0)0, NHCH,CH,O0, a bond, and C(=0)N(R?"), with each group being drawn with its left end attached to G and its right end attached to the carbon substituted with R?' and R%: when present, K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; when present, L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring; nis 0, 1,2, or 3; rR?! R%, R? ,R* R%, and R® are independently selected from hydrogen, C;- Cs alkyl optionally substituted with an imaging moiety, and an imaging moiety; and Y is selected from a bond, carbon, and oxygen; provided that when Y is a bond. K and L are absent and M is selected from arvl and heteroaryl: and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl. aryl, C,-Cs alkyl optionally substituted with an imaging moiety, and heteroaryl: provided that at least one imaging moiety is present in formula (II).

12. The contrast agent of claim 11, wherein R% is C,-Cq alkyl wherein the C,-Cs alkyl is tert-butyl.

13. The contrast agent of claim 11, wherein R® is C,-Cq alkyl wherein the C;-Cq alkyl is methyl. AMENDED SHEET

PCT/US2005/004687

(

14. The contrast agent of claim 1 of formula (III) Oo RA i R?*' R 23 N R n R* K R26 / R25 A (11D), wherein:

J is selected from N(R”), S, O, C(=0), C(=0)0O, NHCH,CH0, a bond, or C(=0)N(R?), with each group being drawn with its left end attached to G and its right end attached to the carbon substituted with R*! and R*;

when present, K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-C¢ alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety;

when present, L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-C¢ alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety;

M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cg alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or

L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring;

Q is halo or haloalkyl:

nis 0, 1,2, or 3;

rR? RR, R¥ R®,R®, and R? are independently selected from hydrogen, C,-C¢ alkyl optionally substituted with an imaging moiety. and an imaging moiety;

R* is C,-Cg alkyl optionally substituted with an imaging moiety; and

Y is selected from a bond, carbon, and oxygen; provided that when Y is a bond, K and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl, aryl, C,-Cg alkyl optionally substituted with an imaging moiety, and heteroaryl;

provided that at least one imaging moiety is present in formula (III).

AMENDED SHEET

PCT/US2005/004687 C

15. The contrast agent of claim 14, wherein J is O and R% is C;-Ce alkyl wherein the C;-Cq alkyl is tert-butyl.

16. The contrast agent of claim 1 wherein the contrast agent is selected from

[0] 0 0 “Oy XX Ci “ry 18

“U . “CL ~ CL 13 | P Ogg , and .

17. The contrast agent of claim 1 of formula (IV): RY R36 R38 R35 R39 Q J R?' R R23 NC 24 N J R / n R28 K R* /

Y. R25 / ~ M (IV), wherein: J is selected from N(R?"), S, O, C(=0), C(=0)0O, NHCH,CH,0, a bond, and C(=0)N(R?"), with each group being drawn with its left end attached to G and its right end attached to the carbon substituted with R*' and R32: when present, K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-C, alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; L is selected from hydrogen, alkoxyalkyl, alkyloxy. aryl, C,-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cg alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring; Q is halo or haloalkyl; nis0, 1,2, or 3;

RZ. RZ R® RR RZ R® RY RZ R¥ R* RY. R® and RY are AMENDED SHEET

PCT/US2005/004687 ( independently selected from hydrogen, C;-Cg alkyl optionally substituted with an imaging moiety, and an imaging moiety; and Y is selected from a bond, carbon, and oxygen, provided that when Y is a bond, K and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen, K and L are absent and M is selected from hydrogen, alkoxyalkyl, aryl, C,-Cg alkyl optionally substituted with an imaging moiety, and heteroaryl; provided that at least one imaging moiety is present in formula (IV).

18. The contrast agent of claim 17, wherein J is C(=0)N(H), and R* is C;-Cs alkyl wherein the C;-Cg alkyl is methyl.

19. The contrast agent of claim 1 wherein the contrast agent is selected from 1 bed cl cl “ “WN N MN N a ( fo) , [0] , 0 s 1g (o]] 74 J \ N H 1g N | H » NA X / / 0 F [o] ’ [eo] ’ ’ {a XX ¢ (o} cl oA ON - ~~ MY EN 5 N N H AO ue and a I of

20. The contrast agent of claim 1 of formula (V) R34 A rR? R R23 R24 n 4) K R26 /

Y. R25 / > M (V), wherein J is selected from N(R?), S, O, C(=0), C(=0)0, NHCH,CH,O, a bond, and C(=O)NR™; AMENDED SHEET

PCT/US2005/004687 @® K is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C;-C¢ alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; when present, L is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; when present, M is selected from hydrogen, alkoxyalkyl, alkyloxy, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, heteroaryl, and an imaging moiety; or L and M, together with the atom to which they are attached, form a three- or four-membered carbocyclic ring; T and U are independently selected from hydrogen, alkoxy, alkoxyalkyl, C;-Cs alkyl optionally substituted with an imaging moiety, halo, and an imaging moiety; or T and U, together with the carbon atoms to which they are attached, form a five- to six-membered aromatic or non-aromatic ring containing zero to two heterotoms selected from oxygen, nitrogen, and sulfur; wherein said ring is optionally substituted with one, two, or three substituents independently selected from C,-C¢ alkyl optionally substituted with an imaging moiety and an imaging moiety; nis 0, 1, 2, or 3; and R%, R%, RB , R%, R%, RS, RY, and R* are independently selected from hydrogen, C,-C¢ alkyl optionally substituted with an imaging moiety, and an imaging moiety; Y is selected from a bond, carbon, and oxygen, provided that when Y is a bond, K and L are absent and M is selected from aryl and heteroaryl; and provided that when Y is oxygen. K and L are absent and M 1s selected from hydrogen. alkoxyalkyl, aryl, C,-Cs alkyl optionally substituted with an imaging moiety, and heteroaryl; provided at least one imaging moiety is present in formula (V).

21. The contrast agent of claim 20 wherein J is O.

22. The contrast agent of claim 20 of formula (VI) AMENDED SHEET

PCT/US2005/004687 _ Re PN h RZ R24 No NAS oO K NN R26 R25 M L (VD), wherein R® , R%, R® , R%, and R* are independently selected from hydrogen, C;-C¢ alkyl optionally substituted with an imaging moiety, and an imaging moiety; provided that at least one imaging moiety is present in formula (VI).

23. The contrast agent of claim 1 wherein the contrast agent is selected from ent Hen J 0 0 o SX P ’ > ’ P ’ o IN 0 o x A yo ~ } J. NO Lr ir 7 YY A, ” OC NEO PN »and Z )

24. A contrast agent of any one of claims | to 23, substantially as herein described with reference to and as illustrated in any of the examples. AMENDED SHEET