ZA200109909B - Method for treating chronic pain using MEK inhibitors. - Google Patents

Description

METHOD FOR TREATING CHRONIC PAIN

USING MEK INHIBITORS

BACKGROUND

The invention features a method for treating chronic pain using MEK inhibitors. Chronic pain includes neuropathic pain, and chronic inflammatory pain.

Abnormality anywhere in a nerve pathway disrupts nerve signals, which in - turn are abnormally interpreted in the brain, causing neuropathic pain.

Neuropathic pain may be, for example, a deep ache, a burning sensation, or hypersensitivity to touch. Diseases or conditions associated with neuropathic pain include, without limitation, diabetic neuropathy, causalgia, plexus avulsion, neuroma, vasculitis, crush injury, viral infections (e.g., herpes virus infection or . . HIV), constriction injury, tissue injury, nerve injury from the periphery to the central nervous system, limb amputation, hypothyroidism, uremia, chronic . alcoholism, post-operative pain, arthritis, back pain, and vitamin deficiencies.

Infections such as herpes zoster (shingles) can cause nerve inflammation : and produce postherpetic neuralgia, a chronic burning localized to the area of .viral infection. Hyperalgesia is when an already noxious stimulus becomes more painful, and allodynia, when a previously non-noxious stimulus becomes painful (such as contact of clothing or a breeze). Reflex sympathetic dystrophy is accompanied by swelling and sweating or changes in local blood flow, tissue atrophy, or osteoporosis. Causalgia, including severe burning pain and swelling, : sweating, and changes in blood flow, may follow an injury or disease of a major nerve such as the sciatic nerve. Some types of chronic low back pain can have a neuropathic component (e.g., sciatica, postpoliomyelitis and CPRM).

Neuropathic pain may also be induced by cancer or chemotherapy.

Neuropathic pain is currently treated with anticonvulsants such as carbamazepine and antidepressants such as amitryptaline. NSAIDS and opioids generally have little effect (Fields et al 1994 Textbook of Pain p 991-996 (pub:

Churchill Livingstone), James & Page 1994 J.Am.Pediatr. Med Assoc, 8: 439-447,

Galer, 1995 Neurology 45 S17-S25. Neuropathic conditions that have been treated with gabapentin include: postherpetic neuralgia, postpoliomyelitis,

CPRM, HIV-related neuropathy, trigeminal neuralgia, and reflex sympathetic dystrophy (RSD). The generally weak efficacy of antiinflammatory agents suggests that the mechanism for chronic pain is separate from hyperalgesia.

SUMMARY OF THE INVENTION

The invention features a method for treating chronic pain, which method includes the step of administering a composition including a MEK inhibitor to a patient in need of such treatment. Chronic pain includes neuropathic pain, idiopathic pain, and pain associated with vitamin deficiencies, uremia, hypothyroidism post-operative pain, arthritis, back pain, and chronic alcoholism. : The invention also features compositions as disclosed, formulated for the treatment of chronic pain. Such a composition may include one or more MEK ; inhibitor compounds having a structure disclosed in patent applications USSN 60/115,652, filed January 13, 1999, USSN 60/115,670, filed January 13, 1999, © 20 USSN 60/115,876 filed January 13, 1999, USSN 60/115,874, PCT/US99/30417, international filing date December 21, 1999, PCT/US99/30418, international filing date December 21, 1999, PCT/US99/30491, international filing date December 21, 1998, and PCT/US99/30435, international filing date December 21, 1999.

RX

Examples of MEK inhibitors include a compound having the formula (1) below:

Ri~o 5

NU!

Ry 52° ’ Re

Ne Rs . ~1

Ry 0)

Ry is H, C 1.3 alkyl, C 3.5 alkenyl, C 35 alkynyl, C 3.3 cycloalkyl, phenyl, (phenyl)C 1.4 alkyl, (phenyl)C 34 alkenyl, (phenyl)C 34 alkynyl, (C 3.4 cycloalkyl)-

C 14 alkyl, (C 3.8 cycloalkyl)C a4 alkenyl, (C 3.5 cycloalkyl)C 14 alkynyl, C 35 heterocyclic radical, (C 3.3 heterocyclic radical)C 1.4 alkyl, (C 3.5 heterocyclic radical)C 34 alkenyl, (C 38 heterocyclic radical)C 34 alkynyl, (CH,).4 OR or (CH2)24 NRcRp. R2is H, C 1.4 alkyl, phenyl, C 35 cycloalkyl, C 3. heterocyclic radical, or (C 3.6 cycloalkyl) methyl. Each of R; and Ry is independently selected ] from H, F, NO2, Br and Cl. Rs is selected from Hand F. Rg is H, F, Cl or CHa.

Each of Rc and Rpis independently selected from H, C 14 alkyl, C 24 alkenyl, C 5. 4 alkynyl, C 36 cycloalkyl, and phenyl; or NRcRp may be a piperidino, morpholino, or N-(C 1.6 alkyl)piperazino ring. Each hydrocarbon radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, hydroxyl, amino, (amino)sulfonyl, and NO,. Each heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 14 alkyl, C 36 cycloalkyl, C 34 alkenyl, C 34 alkynyl, phenyl, hydroxyl, amino, (amino)sulfonyl, and NO, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C 1.2 alkyl, hydroxyl, amino, and

NO,. The invention also includes a pharmaceutically acceptable salt or C 1.5 ester of a disclosed compound. For example, the disclosed alcohol compounds may form esters having the structure obtained by replacing the H of a hydroxyl group with a —-C(=0)C 1.7 acyl group.

The invention also relates to a pharmaceutical composition including

(a) a compound of formula (1) and (b) a pharmaceutically-acceptable carrier.

The invention also features the use of compounds of formulae (Il)A below, such as formula (1A: w.__O uy Rs

I go!

R; I

Ry, X Ry Rg

Rs (IhA : WwW __0 R, . pny R ox 1!

Lo 1

R, > TRIS

X Ry

R,—

A Ry (OA

In formulae (HA and (IDA, Wis OR;, NR.OR;,, NRARs, NR,NRARg, or NR2(CH2)24

NRaRg. Rj is H, C 1. alkyl, C 3.5 alkenyl, C 3g alkynyl, C 3.5 cycloalkyl, phenyl, (phenyl)C 14 alkyl, (phenyl)C 34 alkenyl, (phenyl)C 34 alkynyl, (C 3. cycloalkyl)

C 14 alkyl, (C 3.5 cycloalkyl)C 34 alkenyl, (C 3 cycloalkyl)C 34 alkynyl, C 3.8 heterocyclic radical, (C 3.5 heterocyclic radical)C 14 alkyl, (C 3.8 heterocyclic _radical)C 34 alkenyl, (C 3.5 heterocyclic radical)C 3.4 alkynyl, or (CH2)24NRARB. R: is H, phenyl, C 14 alkyl, Cs4 alkenyl C 3. alkynyl, C 3.4 cycloalkyl, or (C s-

cycloalkyl)C 14 alkyl. Rais H, C 15 alkyl, C 3.5 alkenyl, C 3.5 alkynyl, C 24 cycloalkyl, phenyl, (C 3.5 cycloalkyl)C 1.4 alkyl, (C 3.8 cycloalkyl)C 34 alkenyl, (C 3.8 cycloalkyl)C 3.4 alkynyl, C 3.3 heterocyclic radical, (C 3.4 heterocyclic radical)C 14 alkyl, (aminosulfonyl)phenyl, [(aminosulfonyt)phenyl]C 1.4 alkyl, (aminosulfonyl)C yealkyl, (aminosulfonyl)C 35 cycloalkyl, or [(aminosulfonyl)C 36 cycloalkyl]C 14 alkyl. Rgis H, C 1g alkyl, C 35 alkenyl, C 1.5 alkynyl, C 35 cycloalkyl, or C 6. aryl.

Rs is halo, NO,, SO;NR; (CH2)24NReRr, SO2NR Rk or (CO)T. Tis C 4.5 alkyl, C a. g cycloalkyl, (NReRF)C 1.4 alkyl, ORF, NR{(CH2)2sNReRF, or NReRr. Rs is H or F;

Rs is H, methyl, halo, or NO;; and Rg is H, methyl, halo, or NO,. In formula (IA,

Aris phenyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl. Each of R; and Rg is independently selected from H, halo, C 44 alkyl, SO2NR; (CH2)2.4NRgRu, (CO)(CH2)24NRgRy, (CO)NR(CH2)24NRGgRH, (CO)O(CH2)24NRgRy, SO2NRgRy, and (CO)NRgRy.

However, where Ar is a pyridyl, each of R; and Rg is H. Each of Re, Rp, Re, Rr, :

Ra, and Ry is independently selected from H, C 14 alkyl, C 3.4 alkenyl, C 34 alkynyl, C 3 cycloalkyl, and phenyl. Each of NRcRp, NReRg, and NRgRu can also be independently morpholinyl, piperazinyl, pyrrolidinyl, or piperadinyl. Each . of Ry and R; is independently H, methyl, or ethyl. Rk is C 14 alkyl, C 34 alkenyl, C 34 alkynyl, C 16 cycloalkyl, or phenyl. Xis O, S, or NH. Finally, each . hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 14 alkyl, C a5 cycloalkyl, C 14 alkenyl, C 14 alkynyl, phenyl, hydroxyl, amino, (amino)sulfonyl, and NO, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C 1.; alkyl, hydroxyl, amino, and NO,. In addition to the above compounds, the invention also provides a pharmaceutically acceptable salt or

C 1.7 ester thereof.

The invention also relates to a pharmaceutical composition including (a) a diarylamine, e.g., of formula (l)A, and (b) a pharmaceutically acceptable carrier.

The invention also features the use of a compound having the formula (1)B below:

Wegz® R 10

H

R4 ReR11 J

Rs (HB

In formula (1)B, W is OR1, NR2OR1, NRaRg, NR2NRARs, O(CH2)14NRARs, or = NR2(CH;)14 NRaRp. Rjis H, C 15 alkyl, C 3 alkenyl, C 1. alkynyl, oo fez C 1g cycloalkyl, phenyl, (phenyl)C 14 alkyl, (phenyl)C 14 alkenyl, (phenyl)-

W410 C34 alkynyl, (C 3.5 cycloalkyl)C 1.4 alkyl, (C 1.5 cycloalkyl)C 14 alkenyl, : (C 38 cycloalkyl)C 3.4 alkynyl, C 34 heterocyclic radical, (C 3g heterocyclic radical) wae © C 14 alkyl, (C 1.8 heterocyclic radical)C 34 alkenyl, or (C 34 heterocyclic radical)C co “d+ a4 alkynyl. Each of R; and Rj is independently H, phenyl, C 14 alkyl, g Zn C 1s alkenyl, C 1.5 alkynyl, C 1.5 cycloalkyl, or (C 1.5 cycloalkyl)C 14 alkyl. Each of

R4 Rs and Rg is independently H, F, Br, Cl, or NO2. Rais H, C 1s alkyl, C 3s alkenyl, C 2.5 alkynyl, C 3.4 cycloalkyl, phenyl, (C 3g cycloalkyl)C 1.4 alkyl, (C 38 cycloalkyl)C 34 alkenyl, (C 3.5 cycloalkyl)C 3.4 alkynyl, C 3.3 heterocyclic radical, (C 1.8 heterocyclic radical)C 14 alkyl, (aminosulfonyl)phenyl, (aminosulfonyl)- phenyl] C 14 alkyl, (aminosulfonyl)C 1 alkyl, (aminosulfonyl)C 1.6 cycloalkyl, or [(aminosulfonyl)C 36 cycloalkyl]C 14 alkyl. Regis H, C 15 alkyl, C 35 alkenyl, C 3 alkynyl, C 3.5 cycloalkyl, or phenyl. J is SRc, ORc, SO2Rc, SORc, SO2NRoRE, C 1.8 alkyl, C 3. alkenyl, C 3.5 alkynyl, C 3.5 cycloalkyl, C ss cycloalkenyl, phenyl, (C 1.5 cycloalkyl)C 14 alkyl, (C 3.5 cycloalkyl)C 34 alkenyl, (C 35 cycloalkyl)-

C 14 alkynyl, C 5. heterocyclic radical (e.g., 1,2,5-thiadiazol-3-yl), .

(C 3.5 heterocyclic radical) C 4.4 alkyl, -M'E'G’, (heterocyclic radical)-M'-E’-G’, or (cycloalkyl)-M'-E’-G’. M'is O, SO, SO,, NRg, (CO)NRg, NRe (CO), SO:NRg,

NReSO;, or CH,. E’ is absent (in other words, a covalent bond), (CH3)44 or (CH2)m O(CH,), where 1 < (each of m and p independently) <3 and2<(m+p)< 5S 4. G'is OR3, SORc, SO2Rc, or NRrRg; provided that where p = 1, then G' is H.

Each of Rc, Rp, Re, Rr and Rg is independently selected from H, C 1 alkyl,

C 14 alkenyl, C 34 alkynyl, C 34 cycloalkyl, C 34 heterocyclic radical, and phenyl;

NRrRg and NRpREe can each also independently be selected from morpholinyl, pyrazinyl, piperazinyl, pyrrolidinyl, or piperadinyl. Rqq is H, C 14 alkyl, halo, NO, or SO;NRuR:. Riis H, halo, or NO,.

Each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo,

C 14 alkyl, C a6 cycloalkyl, C 34 alkenyl, C 34 alkynyl, phenyl, hydroxy, amino, (amino)sulfonyl, and NO2, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C 4.; alkyl, hydroxy, amino, and

NO.. The invention also encompasses a pharmaceutically acceptable salt or C 4. 7 ester of a compound of formula (1)B.

The invention also relates to a pharmaceutical composition including (a) a compound of formula (1)B and (b) a pharmaceutically-acceptable carrier.

The invention also features the use of a compound having the formula (1)C below: w_ _O

HOO

RR, ! (nc

W is OR;, NR2OR;, NRaRs, NRoNRaRg, or NRy(CH2)24 NRaRgs. Riis H, C 1 alkyl, C 3.3 alkenyl, C 33 alkynyl, C 34 cycloalkyl, phenyl, (phenyl)C 14 alkyl, (phenyl)C 3.4 alkenyl, (phenyl)C 3.4 alkynyl, (C 1.3 cycloalkyl)-C 1.4 alkyl, (C 3.6 cycloalkyl)C 34 alkenyl, (C 3.3 cycloalkyl)C 34 alkynyl, C 3.5 heterocyclic radical, (C 1g heterocyclic radical)C 1.4 alkyl, (C 3.5 heterocyclic radical)C 3.4 alkenyl, (C 35 heterocyclic radical)C 3.4 alkynyl, or (CH,)24NRaRs. Rz is H, phenyl, C 44 alkyl,

Cs4 alkenyl C 35 alkynyl, C 35 cycloalkyl, or (C 3.3 cycloalkyl)C 14 alkyl. Rais H, C 1-6 alkyl, C 3.5 alkenyl, C 35 alkynyl, C 35 cycloalkyl, phenyl, (C s.s cycloalkyl)C 1.4 alkyl, (C a cycloalkyl)C 34 alkenyl, (C 3.4 cycloalkyl)C 34 alkynyl, C 3 heterocyclic radical, (C 3. heterocyclic radical)C 1.4 alkyl, (aminosulfonyl)phenyl, [(aminosulfonyl)phenyl]C 1.4 alkyl, (aminosuifonyl)C 15 alkyl, (aminosulfony!)C 36 : cycloalkyl, or [(aminosulfonyl)C 3. cycloalkyl]C 14 alkyl. Rg is H, C 1.5 alkyl, C 35 alkenyl, C 3.3 alkynyl, C 34 cycloalkyl, or C gg aryl. Rzis H, F, Cl, Br, or NO2. R4 isHorF. Rsis H, methyl or Cl. Reis H, C 14 alkyl, hydroxyethyl, hydroxypropyl, + (CH2)24(NR¢Rp), phenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl or CHzAr, where Ar is = phenyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl. R;is H, Cy. alkyl, hydroxyethyl, hydroxypropyl, (CH2)24(NRcRp), phenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, or CHAr , where Ar is phenyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl. Each of Rc and Rp is 20» independently selected from H, C 1 alkyl, C 34 alkenyl, C 24 alkynyl, C 35 cycloalkyl, C 3¢ heterocyclic radical, and phenyl. NRcRp can also be selected from morpholinyl, piperazinyl, pyrrolidinyl, or piperadinyl. Each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 14 alkyl, C 3 cycloalkyl, C 24 alkenyl, C 24 alkynyl, phenyl, hydroxy, amino, (amino)sulfonyl, and NO», wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo,

C 1.2 alkyl, hydroxy, amino, and NO,. The invention also features pharmaceutically acceptable salts and C 4.7 esters thereof.

Preferred compounds include PD 297764, 3,4-Difluoro-2-(4-iodo- phenylamino)-N-methoxy-5-(4-pyridin-2-yl-piperazine-1-sulfonyl)-benzamide;

PD 297765, N-Allyloxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-(4-methyl- piperazine-1-sulfonyl)-benzamide; PD297766, N-Allyloxy-5-[(2-diethylamino- ethyl)-methyl-sulfamoyt}-3,4-difluoro-2-(4-iodo-phenylamino)-benzamide;

PD297767, N-Allyloxy-5-[(3-dimethylamino-propyl)-methyl-sulfamoyi]-3,4-difluoro- 2-(4-iodo-phenylamino)-benzamide; PD297768, N-Cyclopropylmethoxy-3,4- difluoro-2-(4-iodo-phenylamino)-5-(4-methyl-piperazine-1-sulfonyl)-benzamide;

PD297769, N-Cyclopropylmethoxy-5-[(2-diethylamino-ethyl)-methyl-sulfamoyi]- 3,4-difluoro-2-(4-iodo-phenylamino)-benzamide; PD297770, N-

Cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-[methyl-(2-pyridin-2- yl-ethyl)-sulfamoyl]-benzamide; PD297771, N-Cyclopropylmethoxy-3,4-difluoro-2- (4-iodo-phenylamino)-5-(4-pyridin-2-yl-piperazine-1-sulfonyl)-benzamide;

PD297772, 5-[Benzyl-(2-dimethylamino-ethyl)-sulfamoyl]-N-cyclopropylmethoxy- 3,4-difluoro-2-(4-iodo-phenylamino)-benzamide; PD297773, 3,4-Difluoro-2-(4- iodo-2-methyl-phenylamino)-N-methoxy-5-(4-pyridin-2-yl-piperazine-1-sulfonyl)- benzamide; and PD297774, 1-[5-Allyloxycarbamoyl-2,3-difluoro-4-(4-iodo-2- methyl-phenylamino)-benzenesulfonyl]-piperidine-3-carboxylic acid amide.

Preferred embodiments of the invention include methods using one or more of the following compounds: (a) said MEK inhibitor has a structure selected from: 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide; and 2-(2- chloro-4-iodo-phenylamino)--cyclopropimethoxy-3,4-difluoro- benzenesulfonamide; (b) said MEK inhibitor has a structure selected from: 2,4-bis-(2-chloro-4-iodo- phenylamino)-3-fluoro-5-nitro-benzoic acid; (c) said MEK inhibitor has a structure selected from: 2-(4-ethynyl-2-methyl- phenylamino)-4-fluoro-benzoic acid; and 2-(3’,5’-dichloro-biphenyl-4-ylamino)- benzoic acid; and (d) said MEK inhibitor has a structure selected from: 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-sulfamoyl-benzamide; 2-(2- chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-5-sulfamoyl-benzamide; C-(2- chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-dimethylsulfamoyl-difluoro- benzamide; N-cyclopropylmethoxy-dimethylsulfamoyl-difluoro-C-(4-iodo-2-

methyl-phenylamino)-benzamide; and C-(2-chloro-4-iodo-phenylamino)-difluoro- (methoxy-methyl-sulfamoyl)-N-(2-morpholin-4-yl-ethoxy)benzamide.

The invention also relates to a pharmaceutical composition including (a) a compound of formula (I)C and (b) a pharmaceutically-acceptable carrier.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a bar graph representing the paw withdrawal threshold (PWT) in grams as a function of time in days. The empty, cross-hatched, and single- hatched bars are vehicle, PD 198306, and pregabalin, respectively. The arrows indicate time of drug administration (30 mg/kg, p.o.).

AS FIG 2. is a bar graph representing the force required in grams to elicit paw i withdrawal using von Frey hair filaments as a function of time in days. Baseline i (BL) measurements were taken before treatment. Animals received a single p.o. administration of PD 198306 (3-30mg/kg), or pregabalin (30mg/kg) and . withdrawal thresholds were re-assessed 1h after treatment. Treatments were £20 repeated twice a day for two days. Results are expressed median + 1% and 3" quartiles. *P<0.05, **P<0.01, ***P<0.001 significantly different from vehicle treated animals (Mann-Whitney t test; n=7-8).

FIG. 3. is a bar graph representing the force required in grams to elicit paw withdrawal using von Frey hair filaments as a function of time in days. Baseline (BL) measurements were taken before treatment. Animals received a single p.o. administration of PD 198306 (3-30mg/kg), or pregabalin (30mg/kg) and withdrawal thresholds were re-assessed 1h after treatment. Treatments were repeated twice a day for two days. Results are expressed median + 1% and 3 quartiles. **P<0.01 significantly different from vehicle treated animals (Mann-

Whitney t test; n=6).

FIG. 4. is a bar graph representing the force required in grams to elicit paw withdrawal using von Frey hair filaments as a function of time in days. Baseline (BL) measurements were taken before treatment. Animals received a single i.t. administration of PD 198306 (1-30ug/10ul), or pregabalin (100ug/10ul) and withdrawal thresholds were re-assessed at 30min, 1h and 2h after treatment.

Results are expressed median + 1* and 3" quartiles. *P<0.05, ***P<0.001 significantly different from vehicle treated animals (Mann-Whitney t test; n=7-9).

FIG. 5. is a bar graph representing the force required in grams to elicit paw withdrawal using von Frey hair filaments as a function of time in days. Baseline (BL) measurements were taken before treatment. Animals received a single it. administration of PD 198306 (1-30ug/10ul), or pregabalin (100ug/10ul) and withdrawal thresholds were re-assessed at 30min, 1h and 2h after treatment.

Results are expressed median + 1% and 3“ quartiles. *P<0.05, **P<0.01, ***P<0.001 significantly different from vehicle treated animals (Mann-Whitney t test; n=6-8).

FIG. 6 is a bar graph representing the force required in grams to elicit paw withdrawal using von Frey hair filaments as a function of time in days . Animals received a single intraplantar (i.pl.) administration of PD 198306 (3mg/100ul), or an intrathecal injection of PD 198306 (30ng/10ul) and withdrawal thresholds were re-assessed 1h after treatment. Results are expressed median + 15 and 3" quartiles. **P<0.01 significantly different from vehicle treated animals (Mann-

Whitney t test; n=6-9).

FIG. 7. is a bar graph representing the force required in grams to elicit paw withdrawal using von Frey hair filaments as a function of time in days. Animals received a single intraplantar (i.pl.) administration of PD 198306 (3mg/100yul), or

’ an intrathecal injection of PD 198306 (30ug/10ul) and withdrawal thresholds were re-assessed 1h after treatment. Results are expressed median + 1% and 3' quartiles. **P<0.01 significantly different from vehicle treated animals (Mann-

Whitney t test; n=6).

FIG. 8 is a bar graph representing the force required in grams to elicit paw withdrawal using von Frey hair filaments. Baseline (BL) measurements were taken before treatment. Animals received a single i.t. administration of

PD219622, PD297447, PD 184352, or PD 254552 (30ug/10ul), or pregabalin (100ug/10ul) and withdrawal thresholds were re-assessed at 30min, 1h and 2h after treatment. Results are expressed median + 1% and 3" quartiles. *P<0.05, **P<0.01, ***P<0.001 significantly different from vehicle treated animals (Mann-

Whitney t test; n=7-8). : " DETAILED DESCRIPTION - The compounds disclosed herein are pharmaceutically active, for example, they inhibit MEK. MEK enzymes are dual specificity kinases involved in, for example, immunomodulation, inflammation, and proliferative diseases such as cancer and restenosis. = Proliferative diseases are caused by a defect in the intracellular signaling system, or the signal transduction mechanism of certain proteins. Defects include a change either in the intrinsic activity or in the cellular concentration of one or more signaling proteins in the signaling cascade . The cell may produce a growth factor that binds to its own receptors, resulting in an autocrine loop, which continually stimulates proliferation. Mutations or overexpression of intracellular signaling proteins can lead to spurious mitogenic signals within the cell. Some of the most common mutations occur in genes encoding the protein known as Ras, a G-protein that is activated when bound to GTP, and inactivated when bound to

GDP. The above-mentioned growth factor receptors, and many other mitogenic receptors, when activated, lead to Ras being converted from the GDP-bound state to the GTP-bound state. This signal is an absolute prerequisite for proliferation in most cell types. Defects in this signaling system, especially in the deactivation of the Ras-GTP complex, are common in cancers, and lead to the signaling cascade below Ras being chronically activated.

Activated Ras leads in turn to the activation of a cascade of serine/threonine kinases. One of the groups of kinases known to require an active Ras-GTP for its own activation is the Raf family. These in turn activate

MEK (e.g., MEK1 and MEK?>) which then activates MAP kinase, ERK (ERK; and

ERK). Activation of MAP kinase by mitogens appears to be essential for proliferation; constitutive activation of this kinase is sufficient to induce cellular transformation. Blockade of downstream Ras signaling, for example by use of a dominant negative Raf-1 protein, can completely inhibit mitogenesis, whether induced from cell surface receptors or from oncogenic Ras mutants. Although

Ras is not itself a protein kinase, it participates in the activation of Raf and other kinases, most likely through a phosphorylation mechanism. Once activated, Raf and other kinases phosphorylate MEK on two closely adjacent serine residues, $218 and $222 in the case of MEK-1, which are the prerequisite for activation of

MEK as a kinase. MEK in turn phosphorylates MAP kinase on both a tyrosine,

Y185, and a threonine residue, T183, separated by a single amino acid.

This double phosphorylation activates MAP kinase at least 100-fold.

Activated MAP kinase can then catalyze the phosphorylation of a large number of proteins, including several transcription factors and other kinases. Many of these

MAP kinase phosphorylations are mitogenically activating for the target protein, such as a kinase, a transcription factor, or another cellular protein. In addition to

Raf-1 and MEKK, other kinases activate MEK, and MEK itself appears to be a signal integrating kinase. Current understanding is that MEK is highly specific for the phosphorylation of MAP kinase. In fact, no substrate for MEK other than the

MAP kinase , ERK, has been demonstrated to date and MEK does not phosphorylate peptides based on the MAP kinase phosphorylation sequence, or even phosphorylate denatured MAP kinase. MEK also appears to associate strongly with MAP kinase prior to phosphorylating it, suggesting that phosphorylation of MAP kinase by MEK may require a prior strong interaction between the two proteins. Both this requirement and the unusual specificity of

MEK are suggestive that it may have enough difference in its mechanism of action to other protein kinases that selective inhibitors of MEK, possibly operating through allosteric mechanisms rather than through the usual blockade of the ATP binding site, may be found.

The effect of the MEK inhibitor PD 198306 has been investigated in two animal models of neuropathic pain by assessing static allodynia with von Frey hairs.

Oral administration of PD 198306 (3-30mg/kg) had no effect in the model of chronic constriction injury of the sciatic nerve (CCl). However, after repeated administration (3 doses over two days) it had a transient effect in the diabetic neuropathy model (streptozocin). This may be due to disorders of the blood-brain barrier induced by the diabetic condition in these animals, thus allowing central action of the compound. Intrathecal administration of PD 198306 (1-30png) dose- dependently blocked static allodynia in both the streptozocin and the CCI models of +, neuropathic pain, with minimum effective doses (MED) of 3 and 10ug respectively. =» The highest dose used (30ug) totally blocked the maintenance of static allodynia, for up to 1h. Intraplantar administration of PD 198306 (3mg/100ul) at a dose 100- - fold higher than the dose shown to be effective intrathecally (30ng/10ul) had no effect on static allodynia in either of the neuropathic pain models. This finding :# confirms the lack of effect seen after systemic administration and suggests a central site of action for the compound.

From this study we can suggest the use of MEK inhibitors as potential new therapeutic tools for chronic pain. The study of potential side-effects, especially related to memory, of future brain-penetrant MEK inhibitors will indicate the therapeutic window for this novel class of compounds in the treatment of pain.

A. Terms

Certain terms are defined below and by their usage throughout this disclosure.

Alkyl groups include aliphatic (i.e., hydrocarbyl or hydrocarbon radical structures containing hydrogen and carbon atoms) with a free valence. Alkyl groups are understood to include straight chain and branched structures.

Examples include methyl, ethyl, propyl, isopropyl, butyl, n-butyl, isobutyl, t-butyl, pentyl, isopentyl, 2,3-dimethylpropyl, hexyl, 2,3-dimethylhexyl, 1,1-dimethylpentyl, : heptyl, and octyl. Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

Alkyl groups can be substituted with 1, 2, 3 or more substituents which are independently selected from halo (fluoro, chloro, bromo, or iodo), hydroxy, amino, : alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy. Specific examples include fluoromethyl, hydroxyethyl, 2,3-dihydroxyethyl, (2- or 3-furanyl)methyl, cyclopropylmethyl, benzyloxyethyl, (3-pyridinyl)ymethyl, (2- or 3-furanyl)methyl, (2-thienyhethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminocbutyl, methoxymethyl, N-pyridinylethyl, diethylaminoethyt, and cyclobutylmethyl.

Alkeny! groups are analogous to alkyl groups, but have at least one double bond (two adjacent sp® carbon atoms). Depending on the placement of a double bond and substituents, if any, the geometry of the double bond may be entgegen (E), or zusammen (Z), cis, or trans. Similarly, alkynyl groups have at least one triple bond (two adjacent sp carbon atoms). Unsaturated alkenyl or alkynyl groups may have one or more double or triple bonds, respectively, or a mixture thereof; like alkyl groups, unsaturated groups may be straight chain or branched, and they may be substituted as described both above for alkyl groups and throughout the disclosure by example. Examples of alkenyls, alkynyls, and substituted forms include cis-2-butenyl, trans-2-butenyl, 3-butynyl, 3-phenyl-2- propynyl, 3-(2'-fluorophenyl)-2-propynyl, 3-methyl(5-phenyl}-4-pentynyl, 2- hydroxy-2-propynyl, 2-methyl-2-propynyl, 2-propenyl, 4-hydroxy-3-butynyl, 3-(3- fluorophenyl)-2-propynyl, and 2-methyl-2-propenyl. In formula (I), alkenyls and alkynyls can be C ,4 or C 2.3, for example, and are preferably C 34 or C 33.

y

R WO 01/05393 PCT/US00/18348

More general forms of substituted hydrocarbon radicals include hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl, and corresponding forms for the prefixes amino-, halo- (e.g., fluoro-, chloro-, or bromo-), nitro-, alkyl-, phenyl-, cycloalkyl- and so on, or combinations of 5S substituents. According to formula (1), therefore, substituted alkyls include hydroxyalkyl, aminoalkyl, nitroalkyl, haloalkyl, alkylalkyl (branched alkyls, such as methylpentyl), (cycloalkyl)alkyl, phenylalkyl, alkoxy, alkylaminoalkyl, dialkylaminoalkyl, arylalkyl, aryloxyalkyl, arylalkyloxyalkyl, (heterocyclic radical)alkyl, and (heterocyclic radical)oxyalkyl. Rj thus includes hydroxyalkyl, 10 hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl, aminoalkyl, aminoalkenyl, aminoalkynyl, aminocycloalkyl, aminoaryl, alkylalkenyl, (alkylaryl)alkyl, (haloaryl)alkyl, (hydroxyaryl)alkynyl, and so forth. Similarly, Ra includes hydroxyalkyl and aminoaryl, and Rg includes hydroxyalkyl, aminoalkyl, and hydroxyalkyl(heterocyclic radical)alkyl. 15 Heterocyclic radicals, which include but are not limited to heteroaryls, = include: furyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, pyrrolyl, imidazolyl, 1,3,4- . triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indolyl, and their nonaromatic counterparts. Further examples of heterocyclic radicals include ] piperidyl, quinolyl, isothiazolyl, piperidinyl, morpholinyl, piperazinyl, 20 tetrahydrofuryl, tetrahydropyrrolyl, pyrrolidinyl, octahydroindolyl, octahydrobenzothiofuranyl, and octahydrobenzofuranyl.

Selective MEK 1 or MEK 2 inhibitors are those compounds which inhibit the MEK 1 or MEK 2 enzymes, respectively, without substantially inhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase, EGF, and PDGF receptor kinases, and C-src. In general, a selective MEK 1 or MEK 2 inhibitor has an ICs for MEK 1 or MEK 2 that is at least one-fiftieth (1/50) that of its ICs for one of the above-named other enzymes. Preferably, a selective inhibitor has an ICs that is at least 1/100, more preferably 1/500, and even more preferably 1/1000, 1/5000, or less than that of its {Cs or one or more of the above-named enzymes.

if

B. Compounds

One aspect of the invention features the use of disclosed compounds shown in formula (1) in the Summary section.

Embodiments of the invention include compounds wherein: (a) Riis bromo or chloro; (b) Rs is fluoro; (¢) Rs is H; (d) each of Rs and Rs is H; (e) each of R4 and R; is fluoro; (f) R3 is bromo; (g) Rj is fluoro; (h) Ry is nitro; (i) Rs is H; (i) Rs is chloro; (k) Re is methyl; (I) Ry is H or C 14 alkyl, and Rz is H; (m) R, is (C 36 cycloalkyl)methyl; (n) Ry is H; (0) Ry is (CH) 240R¢ or (CH2) 2.4 NRcRp; (p) Rs is chloro or methyl; (q) Re is H; or combinations thereof.

Preferably, when R4, Re, or Rp is an alkenyl or alkynyl, the double or triple bond, respectively, is not adjacent the point of attachment when the point of attachment is a heteroatom. For example, R; is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferably prop-1-ynyl or but-1-enyl.

Examples of compounds of formula (I) include: 4-fluoro-2-(4-iodo-2- methyl-phenylamino)-benzenesulfonic acid; 4-fluoro-N-hydroxy-2-(4-iodo-2- methyl-phenylamino)-benzenesuifonamide; N-cyclopropylmethoxy-4-fluoro-2-(4- iodo-2-methyl-phenylamino)-benzenesulfonamide; 3,4-difluoro-2-(4-iodo-2- methyl-phenylamino)-benzenesulfonic acid; 3,4-difluoro-N-hydroxy-2-(4-iodo-2- methyl-phenylamino)-benzenesulfonamide; N-cyclopropyimethoxy-3,4-difluoro-2- (4-iodo-2-methyl-phenylamino)-benzenesulfonamide; 3,4,5-trifluoro-2-(4-iodo-2- methyl-phenylamino)-benzenesulfonic acid; 3,4,5-trifluoro-N-hydroxy-2-(4-iodo-2- methyl-phenylamino)-benzenesuifonamide; N-cyclopropylmethoxy-3,4,5-trifluoro- 2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide; 5-bromo-3,4-difluoro-2-(4- iodo-2-methyl-phenylamino)-benzenesulfonic acid; 5-bromo-3,4-difluoro-N- hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide; 5-bromo-N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)- benzenesulfonamide; 2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzenesuifonic acid; N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzenesulfonamide; or

N-cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro- benzenesulfonamide.

»

Further examples of compounds include: 2-(2-chloro-4-iodo- phenylamino)-4-fluoro-benzenesulfonic acid; 2-(2-chloro-4-iodo-phenylamino)-4- fluoro-N-hydroxy-benzenesulfonamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-4-fluoro-benzenesulfonamide; 2-(2-chloro-4-iodo- phenylamino)-3,4-difluoro-benzenesutfonic acid; 2-(2-chloro-4-iodo- phenylamino)-3,4-difluoro-N-hydroxy-benzenesulfonamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide; 2-(2- chloro-4-iodo-phenylamino)-3,4,5-trifluoro-benzenesulfonic acid; 2-(2-chloro-4- iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzenesulfonamide; 2-(2-chloro-4- iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzenesulfonamide; 5- bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-diflucro-benzenesulfonic acid; 5- bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy- benzenesulfonamide; 5-bromo-2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide; 2-(2-chloro-4-iodo- phenylamino)-4-nitro-benzenesulfonic acid; 2-(2-chloro-4-iodo-phenylamino)-N- = + hydroxy-4-nitro-benzenesulfonamide; or 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-4-nitro-benzenesulfonamide.

A second aspect of the invention features the use of compounds shown in formulae (1)A and (IIA in the Summary section. Embodiments of the invention * includes compounds of formula ()A wherein: (a) Rs is NO; (b) Rq is fluoro; (c) each of Rs and R, is independently selected from H and fluoro; (d) Rs is methyl, fluoro, or chloro; (e) Re is methyl, chloro, fluoro, nitro, or hydrogen; (f) Re is H; (g) Re is fluoro; (h) Rk is methyl or ethyl; (i) Ry is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenyl, phenethyl, allyl, C 1.5 alkenyl, C 16 cycloalkyl, (C 3.5 cycloalkyl)C 4.5 alkyl, (C 2.5 heterocyclic radical)C 1.2 alkyl, or (CHz)24

NRcRp; (j) Ry is H or (C 3.4 cycloalkyl)C 1.» alkyl; (k) R; is H or methyl; (I) Ra has at least one hydroxyl substituent; (m) Ra is H, methyl, ethyl, isobutyl, hydroxyethyl, phenyl, 2-piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2- hydroxyethyl)-piperazin-1-yll-propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino- ethyl; and Rg is H; or where Rg is methyl and Ra is phenyl; (n) Wis NRaRg or

NR2NRaRsg; (0) W is NR2(CHa),.4 NRaRg or O(CHa)2.3 NRaRg; (p) W is NR2ORj;

Wf (q) Wis OReg; (r) Ry is in the para position relative to X; (s) R; is iodo; (t) Rg is in the ortho position relative to X; (u) or combinations thereof.

In additional embodiments, if Rs is H, then Rs is nitro; or Rg is methyl, halo, or nitro; or R3 is SO2NR, (CH2)24NRegRg, SO2NR| Rk or (CO)T. In some embodiments, Aris phenyl (e.g., formula (1)A), and in other embodiments, Ar is 2- pyridyl, 3-pyridyl, or 4-pyridyl. Preferably, where one of Ry, Raz, Ra, Rs, Re, and Rp is an alkenyl or alkynyl group, the double or triple bond, respectively, is not adjacent the point of attachment. For example, where W is NR2OR;y, Rz is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferably prop-1-ynyl or but- 1-enyl. Some embodiments include the formula 2,4-bis-(2-chloro-4-iodo- phenylamino)-3-fluoro-5-nitro-benzoic acid, the compounds in the following list, and 2-methyl (instead of 2-chloro) analogs thereof. 1. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-(4-sulfamoyl- phenylamino)-benzoic acid; 2. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-phenylamino-benzoic acid; 3. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-phenoxy-benzoic acid; 4. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-phenylsulfanyl-benzoic acid; ’ 5. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-4-(methyl-phenyl-amino)-5-nitro- benzoic acid; 6. 2-{(2-chloro-4-iodophenyl)amino}-3-fluoro-4-[[4-[[(2-hydroxyethyl)amino}- carbonyl]phenyl]amino]-5-nitro-benzoic acid, 7. 2-[(2-chloro-4-iodophenyl)amino]-4-[[4-[(dimethylamino)carbonyl} phenyllamino]-3-fluoro-5-nitro-benzoic acid, 8. 2-(2-Chloro-4-iodo-phenylamino)-3,5-difluoro-4-phenylamino-benzoic acid; 9. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-(3-sulfamoyl- phenylamino)-benzoic acid; 10. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-(2-sulfamoyl- phenylamino)-benzoic acid;

A}

N WO 01/05393 PCT/US00/18348 11. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hyd roxy-5-nitro-4-(4- sulfamoyl-phenylamino)-benzamide; 12. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hydroxy-5-nitro-4- phenylamino-benzamide; 13. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hydroxy-5-nitro-4-phenoxy- benzamide; 14. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hydroxy-5-nitro-4- phenylsulfanyl-benzamide; 15. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hydroxy-4-(methyl-phenyi- ‘amino)-5-nitro-benzamide; 16. 2-[(2-chloro-4-iodophenyt)amino]-3-fluoro-N-hydroxy-4-[{4-[[(2- hydroxyethyl)amino]-carbonyl]phenylJamino}-5-nitro-benzamide; 17. 2-[(2-chloro-4-iodophenyl)amino]-4-[[4-[(dimethylamino)carbonyl] phenyl]lamino]-3-fluoro-N-hydroxy-5-nitro-benzamide; 18. 2-(2-Chloro-4-iodo-phenylamino)-3,5-difluoro-N-hydroxy-4-phenylamino- te benzamide; 19. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hydroxy-5-nitro-4-(3- sulfamoyl-phenylamino)-benzamide; a 20. 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-N-hydroxy-5-nitro-4-(2- sulfamoyl-phenylamino)-benzamide; = 21. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-5-nitro-4- (4-sulfamoyl-phenylamino)-benzamide; 22. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-5-nitro-4- phenylamino-benzamide; 23. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-5-nitro-4- phenoxy-benzamide; 24. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-5-nitro-4- phenylsulfanyl-benzamide; 25. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-4- (methyl-phenyl-amino)-5-nitro-benzamide; 26. 2-[(2-chloro-4-iodophenyl)amino]-3-fluoro-N-cyclopropyimethoxy-4-{[4-{[(2- hydroxyethyl)amino]-carbonyl]phenyl]amino]-5-nitro-benzamide;

tr 27. 2-[(2-chloro-4-iodophenyl)amino}-4-[[4-[(dimethylamino)carbonyl]phenyl]- amino]-3-fluoro-N-cyclopropylmethoxy-5-nitro-benzamide; 28. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,5-difluoro-4- phenylamino-benzamide; 29. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-5- nitro-4-(3-suifamoyl-phenylamino)-benzamide; and 30. 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3-fluoro-5-nitro-4- (2-suifamoyl-phenylamino)-benzamide.

In the scheme below, W can also be any of the values described herein for formula (I)A or (II)A in the section describing preferred values for W. The compound numbers provided in the scheme correspond to the numbers provided in the above list; these compounds are illustrative, not limitative, of the invention.

»

FORMULAE (l)A AND (IlI)A EXAMPLES w__0 w__0 uy wu © 1 N 6 N

Cmpd # (W) Cmpd # (W)

I (OH) 6 (OH)

ON F I 11 (NHOH) ON F I 16 (NHOH)

H 21 (NHOCH,"Pr) H 26 (NHOCH, Pr)

OU HC

Neh, . 0) 2 wW__O w__0 . y © 7 I

N N ot Crmpd # (W) TL Cmpd # (W) 2 (0H) 7 (OH)

ON F LP) (NHOH) ON F ! 17 (NHOH)

H 22 (NHOCH, Pr) H 27 (NHOCH, Pr) py ty /4 ~~

ANH OH

0] 0} 3 8 wW__O w__O nu § nw ©

N Cmpd # (W) N Cmpd # (W) 3 (OH) 2 (OH)

ON E | 13(NHOH) F F I 18 (NHOH) _

S 23 (NHOCH, Pr) H 28 (NHOCH,°Pr) 4 wW__O La wW__0 Loa

N 9 N

Cmpd # (W) Cmpd # (W) 4 (OH) 9 (OH)

O,N F I 14 (NHOH) ON F I 19 (NHOH)

UO 24 (NHOCH, Pr) H OC 29 (NHOCH, Pr) 0=5=0

NH

W_0 La 10 W_ 0 a

N Cmpd # (W) N Cmpd # (W) 5 (OH) 10 (OH) (NHOH) 20 (NHOH)

ON A F boos (NHOCH, Pr) ON . F ! 30 (NHOCH, Pr)

H,C’ 0,

HN lo}

[J

A third aspect of the invention features the use of compounds shown in formula (I)B in the Summary section. Embodiments of the invention include compounds wherein: (a) Re is C 1.2 alkyl; (b) Wis OH, or Wis NHOR, (c) Ry is methyl or chloro; (d) R44 is fluoro; (e) Ry4 is H; (f) J is trihalomethy! or methyithio; (9) J is SO,CHs3; (h) J is SOCHGg; (i) J is C 3.5 alkynyl where the triple bond is between the carbon atoms alpha and beta to the phenyl group; (j) Ry has at least one hydroxy substituent; (k) R, is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 35 alkenyl, C 3.5 alkynyl, C 36 cycloalkyl, (C 3.5 cycloalkyl)C 4; alkyl, or (C 3.5 heterocyclic radical)C 4.2 alkyl; (I) Ry is H or (C 34 cycloalkyl)C 1.; alkyl; (m) Rz is H, methyl, C 34 alkynyl, C 3.5 cycloalkyl, or (C 1.5 cycloalkyl)methyl; (n) Ra is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C 14 alkynyl, phenyl, 2-piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-{4-(2-hydroxyethyl)-piperazin-1-yi]-propyl, 2-pyrrolidin-1- 1S vyl-ethyl, or 2-diethylamino-ethyl; and Rg is H; or where Rg is methyl and Ra is phenyl; (0) each of Rs and Rg is H, and Rs is F; (p) each of R4, Rs, and Rg is F; (q)

Rs is F; (r) each Rs and Rg is F and Rg is Br; (s) each Rs and Rg is F and Rg is H; (t) J is 1,2,5-thiadiazol-3-yl; or a combination thereof.

Preferably, where one of Ry, Rz, Ra, Rs, Re, Rp, Re, Rr, and Rg, for example, is an alkenyl or alkynyl group, its double or triple bond, respectively, is not adjacent the point of attachment. For example, where W is NR,OR;, Rz is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferably prop-1-ynyl or but- 1-enyl.

Examples of compounds of formula (1)B include: 4-fluoro-2-(2-methyi-4- methylsulfanyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(2-methyl-4- methyisulfanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4-methanesulfinyl-2- methyl-phenylamino)-benzoic acid; 2-(4-methanesulfinyl-2-methyl-phenylamino)- 4-nitro-benzoic acid; 3,4,5-trifluoro-2-(4-methanesulfonyl-2-methyl-phenylamino)- benzoic acid; 3,4-difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 2-(2-methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzoic acid; 3,4,5- trifluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 4-fluoro-2-(4- methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(4-

] methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4,5-trifluoro-2-(2-methyl- 4-methylsulfanyl-phenylamino)-benzoic acid; 4-fluoro-2-(4-methane-sulfinyl-2- methyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(4-methanesulfinyl-2- methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4-methanesulfonyl-2-methyl- phenylamino)-benzoic acid; and 2-(4-methanesulfonyl-2-methyl-phenylamino)-4- nitro-benzoic acid; and the corresponding hydroxamic acid or . cyclopropylhydroxamic acid of each.

Preferred examples of compounds of formula (1)B are : 4-Fluoro-2-(4- methanesulfanyl-phenylamino)-benzoic acid (1); 4-Fiuoro-2-(4-methanesulfinyl- phenylamino)-benzoic acid (2); 4-Fluoro-2-(4-methanesulfonyl-phenylamino)- benzoic acid (3); 4-Fluoro-2-(2-methyl-4-trimethylsilanylethynyl-phenylamino)- benzoic acid (6); 4-Fluoro-2-(2-methyl-4-ethynyl-phenylamino)-benzoic acid (7).

Biological data on these seven compounds is given on page 17; full characterization of the compounds - MP, NMR, MS, IR and CHN- is given on pages 28-31. . Additional preferred compounds include the following: (a) 5-Bromo-2-(4- ~ ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy- 2-(4-ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzamide; 2-(4-Ethynyl-2- . methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4- ethynyl-2-methyl-phenylamino)-3,4,5-trifluoro-benzamide; 2-(4-Ethynyl-2-methyl- ~ phenylamino)-3,4,5-trifluoro-benzoic acid; 5-Bromo-N-cyclopropylmethoxy-2-(4- ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzamide; (b) 5-Bromo-2-(4-ethynyl-

Cl-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4- ethynyl-Cl-methyl-phenylamino)-3,4-difluoro-benzamide; 2-(4-Ethynyl-Cl-methyl- phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4-ethynyl-Cl- methyl-phenylamino)-3,4,5-trifluoro-benzamide; 2-(4-Ethyny!-Cl-methyl- phenylamino)-3,4,5-trifluoro-benzoic acid; 5-Bromo-N-cyclopropylmethoxy-2-(4- ethynyl-Cl-methyl-phenylamino)-3,4-difluoro-benzamide; (c) 5-bromo-3,4-difluoro- 2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4- methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 3,4,5-trifluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(4-

[J methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzoic acid; (d) 5-bromo-N- cyclopropylmethoxy-3,4-difluoro-2-(2-methyl-4-methylsulifanyl-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfinyl-2-methyi- phenylamino)-benzamide; N-cyclopropylmethoxy-3,4,5-trifluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4- difluoro-2-(2-methyl-4-methyisulfanyl-phenylamino)-benzamide;

N-cyclopropylmethoxy-3,4,5-trifluoro-2-(4-methanesulfinyl-2-methyl- phenylamino)-benzamide; 5-bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy- 3,4,5-trifluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; N- cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfonyl-2-methyl-phenylamino)- benzamide; (e) N-cyclopropylmethoxy-3,4-difluoro-2-(4-imidazol-1-yl-2-methyl- phenylamino)-benzamide,; (f) N-cyclopropylmethoxy-3,4,5-trifluoro-2-(2-methyl-4- [1,2,5]thiadiazol-3-yl-phenylamino)-benzamide; 2-[4-(4-chloro-[1,2,5]thiadiazol-3- yl)-2-methyl-phenylamino]-3,4,5-trifluoro-benzoic acid; 2-[4-(4-chloro- . [1,2,5]thiadiazol-3-yl)-2-methyl-phenylamino]-N-cyclopropylmethoxy-3,4,5- trifluoro-benzamide; (g) 2-{4-[4-(2-dimethylamino-ethoxy)-[1,2,5]thiadiazol-3-yl}-2- methyl-phenylamino}-3,4,5-trifluoro-benzoic acid; (h) N-cyclopropylmethoxy- 3,4,5-trifluoro-2-{2-methyl-4-[4-(2-piperidin-1-yl-ethoxy)-[1,2,5]thiadiazol-3-yl}- phenylamino}-benzamide.

Further preferred compounds include: (a) 5-bromo-2-(2-chloro4- methyisulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-chloro-4- methanesulfinyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-chloro-4- methanesulfonyl-phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-chloro- methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 5-bromo-2-(2-chloro-4- methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chioro-4- methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; (b) 5-bromo-2-(2- chloro-4-methylsulfanyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(2-chloro-4-methanesulfinyl-phenylamino)-N-cyclopropylmethoxy- 3,4-difluoro-benzamide; 2-(2-chloro-4-methanesulfonyl-phenylamino)- N- cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-(2-chloro-4-methyisulfanyl-

L" phenylamino)- N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4- ~ methanesulfinyl-phenylamino)- N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 5-bromo-2-(2-chloro-4-methanesulfonyl-phenylamino)-N-cyclopropylmethoxy-3,4- difluoro-benzamide; 2-(2-chloro-4-methylsulfanyl-phenylamino)-N- cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-(2-chloro-4-methanesulfonyl- phenylamino)-N-cyclopropyimethoxy-3,4-difluoro-benzamide; and (c) 2-[2-chloro 4-(3H-imidazol-1-yl)-phenylamino]-N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-[1,2,5]thiadiazol-3-yl-phenylamino)-N-cyclopropylmethoxy-3,4,5- trifluoro-benzamide; 2-[4-(2-chloro-4-chloro-[1,2,5]thiadiazol-3-yl)-phenylamino}- 3,4,5-trifluoro-benzoic acid; 2-[2-chloro-4-(4-chloro-[1,2,5]thiadiazol-3-yl)- phenylamino]-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-{4-[4-(2- dimethylamino-ethoxy)-[1,2,5}thiadiazol-3-yl}-2-methyl-phenylamino}-3,4,5- trifluoro-benzoic acid; 2-{2-chioro-4-[4-(2-piperidin-1-yl-ethoxy)-[1,2,5]thiadiazol-3- yl}-phenylamino}-N-cyclopropyimethoxy-3,4,5-trifluoro-benzamide;

Additional preferred compounds include: (a) 2-(2-Chloro-4-ethynyl- phenylamino)-4-fluoro-benzoic acid; 5-Bromo-2-(2-chloro-4-ethynyl- + phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro-4-ethynyl-phenylamino)- N- - cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)- : N-cyclopropylmethoxy-4-nitro-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)- N-hydroxy-3,4,5-trifluoro- benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)-3,4- -difluoro-benzoic acid; 2-(4-Ethynyl-2-chloro-phenylamino)-4-nitro-benzoic acid; 2- (2-Chloro-4-ethynyl-phenylamino)- N-Cyclopropylmethoxy-3,4,5-trifluoro- benzamide; 2-(2-chloro-4-methanesulfinyl-phenylamino)- 4-fluoro-N-hydroxy- benzamide; 5-Bromo-2-(4-ethynyl-2-chloro-phenylamino)-3,4-difluoro-N-hydroxy- benzamide; (b) 2-(2-Chloro-4-ethynyl-phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-Chloro-4-ethynyl-phenylamino)- N-cyclopropylmethoxy-4-fluoro-benzamide; 5-Bromo-2-(2-chloro-4-ethynyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(4-Ethynyl-2-chloro-phenylamino)-3,4-difluoro-N-hydroxy- benzamide; 2-(4-Ethynyl-2-chloro-phenylamino)-N-hydroxy-4-nitro-benzamide; and (c) 2-(2-Chloro-4-ethynyl-phenylamino)-4-fluoro-benzoic acid; 2-(2-Chloro-4- ethynyl-phenylamino)- N-cyclopropylmethoxy-4-fluoro-benzamide; 2-(2-Chloro-4-

[ methanesulfinyl-phenylamino)- 4-fluoro-N-hydroxy-benzamide; 2-(2-chloro-4- imidazol-1-yl-phenylamino)- 3,4-Difluoro-benzoic acid.

A fourth aspect of the invention features the use of compounds shown in formula (I)C in the Summary section.

Examples of compounds of formula (I)C have structures wherein: (a) the sulfamoyl! group is meta to W(CO)- and para to the bridging NH; (b) the sulfamoyl group is para to W (CO)- and meta to the bridging NH; (c) Ras is fluoro; (d) Ra is fluoro; (e) Rs is H; (f) Wis OH; (g) W is NR,ORy; (h) each of R; and R; is fluoro; (i) Ry has at least one hydroxy substituent; (k) Ry is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 2.5 alkenyl, C 2.5 alkynyl,

C 36 cycloalkyl, (C 3.5 cycloalkyl)C 1.2 alkyl, (C 3.5 heterocyclic radical)-

C 1.2 alkyl, or (CH3)24sNRARs; (I) R41 is H or (C 3.4 cycloalkyl)C 4. alkyl; (m) R; is H, methyl, C 24 alkynyl, C 3.5 cycloalkyl, or (C as cycloalkyl)methy!; (n) Ra is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C 14 alkynyl, phenyl, 2-piperidin-1-yl-ethyl, 2,3-dihydroxy- © propyl, 3{4-(2-hydroxyethyl)-piperazin-1-yl]-propyl, 2-pyrrolidin-1-yi-ethyl, or 2- diethylamino-ethyl; and Rg is H; or where Rg is methyl and Ra is phenyl; (0) R7 is (CH2)2.4(NRcRp); (p) NRcRp is selected from morpholinyl, piperazinyl, pyrrolidinyl, or piperadinyl; (9) Rc is methyl, ethyl, hydroxyethyl, or hydroxypropyl; (r) Rs is methyl or chloro; (s) Rp is methyl, ethyl, hydroxyethyl, or hydroxypropyl; (t) or combinations thereof, such as wherein each of Rc and Rp is methyl or ethyl.

Preferably, where one of Ry, Ra, Ra, Rs, Re, or Rp is an alkenyl or alkynyl group, the double or triple bond, respectively, is not adjacent the point of attachment. For example, where W is NR2OR;, R: is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferably prop-1-ynyl or but-1-enyl.

Examples of compounds of formula (1)C include: 2-(2-chloro-4-iodo- phenylamino)-4-sulfamoyl-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-N- hydroxy-4-sulfamoyl-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-4-sulfamoyl-benzamide; 2-(2-chloro-4-iodo-phenylamino)-4- (2-morpholin-4-yl-ethylsulfamoyl)-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-

N-hydroxy-4-(2-morpholin-4-yl-ethylsulfamoyl)-benzamide; 2-(2-chloro-4-iodo-

phenylamino)-N-cyclopropylmethoxy-4-(2-morpholin-4-yl-ethylsulfamoyl)- benzamide; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5-sulfamoyl-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-5-sulfamoyl- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- 5-sulfamoyl-benzamide; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5-(2- morpholin-4-yl-ethylsulfamoyl)-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-3,4- difluoro-N-hydroxy-5-(2-morpholin-4-yl-ethylsulfamoyl)-benzamide; and 2-(2- chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-(2-morpholin-4- yl-ethylsulfamoyl)-benzamide.

Other examples include 5-[bis-(4-methoxy-benzyl)-sulfamoyl}-2-(2-chloro- 4-iodo-phenylamino)-3,4-difluoro-benzoic acid; and 2-(2-chloro-4-iodo- phenylamino)-5-dimethylsulfamoyl-3,4-difluoro-benzoic acid methyl ester.

Additional examples include 5-(bis-pyridin-3-yimethyl-suifamoyl)-3,4- difluoro-2-(4-iodo-phenylamino)-benzoic acid; 5-(bis-pyridin-3-yimethyl- 15. sulfamoyl)-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)- - benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenytamino)-5-(methyl- + pyridin-3-yimethyl-sulfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4- iodo-phenylamino)-5-{(pyridin-3-ylmethyl)-sulfamoyl]-benzamide; N- . cyclopropylmethoxy-5-[(3-diethylamino-propyl)-pyridin-3-ylmethyl-sulfamoyl}-3,4- difluoro-2-(4-iodo-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-5- 5 [(3-hydroxy-propyl)-pyridin-3-yimethyl-sulfamoyl]-2-(4-iodo-phenylamino)- benzamide; N-cyclopropylmethoxy-5-(ethyl-pyridin-3-ylmethyi-sulfamoyl)-3,4- difluoro-2-(4-iodo-phenylamino)-benzamide; N-cyclopropy!methoxy-3,4-difluoro-5- [(2-hydroxy-ethyl)-pyridin-3-ylmethyl-sulfamoyl]-2-(4-iodo-phenylamino)- benzamide; 5-(bis-pyridin-2-ylmethyl-sulfamoy!)-3,4-difluoro-2-(4-iodo- phenylamino)-benzoic acid; 5-(bis-pyridin-2-ylmethyl-sulfamoyl)-N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-benzamide; N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-(methyl-pyridin-2- yimethyl-sulfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- phenylamino)-5-[(pyridin-2-ylmethyl)-sulfamoyl]-benzamide; 5-(bis-pyridin-3- ylmethyl-sulfamoyl)-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid;

* 5-(bis-pyridin-3-ylmethyl-sulfamoyl)-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- 2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- 2-methyl-phenylamino)-5-(methyl-pyridin-3-ylmethyl-sulfamoyl)-benzamide;

N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{(pyridin-3- ylmethyl)-sulfamoyl]-benzamide; N-cyclopropylmethoxy-5-[(3-diethylamino- propyl)-pyridin-3-ylmethyl-sulfamoyl}-3,4-difluoro-2-(4-iodo-2-methyl- phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-5-[(3-hydroxy- propyl)-pyridin-3-ylmethyl-sulfamoyl]-2-(4-iodo-2-methyl-phenylamino)- benzamide; N-cyclopropylmethoxy-5-(ethyl-pyridin-3-ylmethyl-sulfamoyl)-3,4- difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4- difluoro-5-[(2-hydroxy-ethyl)-pyridin-3-ylmethyl-suifamoyl]-2-(4-iodo-2-methyl- phenylamino)-benzamide; 5-(bis-pyridin-2-ylmethyl-sulfamoyl)-3,4-difluoro-2-(4- iodo-2-methyl-phenylamino)-benzoic acid; 5-(bis-pyridin-2-ylmethyl-sulfamoyi)-N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide; N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(methyl- pyridin-2-ylmethyl-suilfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4- iodo-2-methyl-phenylamino)-5-[(pyridin-2-ylmethyl)-sulfamoyl]-benzamide; 5-(bis- pyridin-3-ylmethyl-sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro- benzoic acid, 5-(bis-pyridin-3-ylmethyl-sulfamoyl)-2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-(methyl- pyridin-3-yimethyl-sulfamoyl)-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-5-{(pyridin-3-ylmethyl)-sulfamoyl}-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-5-[(3-diethylamino- propyl)-pyridin-3-ylmethyl-sulfamoyl]-3,4-difluoro-benzamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-{(3-hydroxy-propyl)-pyridin-3- ylmethyl-sulfamoyl]-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-5-(ethyl-pyridin-3-ylmethyl-suifamoyl)-3,4-difluoro- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- 5-[(2-hydroxy-ethyl)-pyridin-3-ylmethyl-sulfamoyl]-benzamide; 5-(bis-pyridin-2- ylmethyl-sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzoic acid; 5- (bis-pyridin-2-ylmethyl-sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-N-

0 cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-5-(methyl-pyridin-2-yimethyl-sulfamoyl)- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- 5-[(pyridin-2-ylmethyl)-sulfamoyl}-benzamide; N-cyclopropyimethoxy-3,4-difluoro- 5-[(3-hydroxy-propyl)-pyridin-2-yimethyl-sulfamoyl]-2-(4-iodo-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-2- yimethyl-sulfamoyl]-2-(4-iodo-phenylamino)-benzamide; 5-(benzyl-pyridin-2- ylmethyl-sulfamoyl)-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5- [(pyridin-4-yimethyl)-sulfamoyl]-benzamide; N-cyclopropylmethoxy-5-(ethyl- pyridin-4-ylmethyi-sulfamoyl)-3,4-difluoro-2-(4-iodo-phenylamino)-benzamide; N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-(methyl-pyridin-4- yimethyl-sulfamoyl)-benzamide; N-cyclopropylmethoxy-3 4-difluoro-5-[(3-hydroxy- propyl)-pyridin-4-yimethyl-sulfamoyl]-2-(4-iodo-phenylamino)-benzamide;

N-cyclopropylmethoxy-3,4-difluoro-5-{(2-hydroxy-ethyl)-pyridin-4-yimethyl- .. sulfamoyl]-2-(4-iodo-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4- . difluoro-2-(4-iodo-phenylamino)-5-(methyl-phenyl-sulfamoyl)-benzamide; N- cyclopropylmethoxy-3,4-diflucro-2-(4-iodo-phenylamino)-5-phenylsutfamoyl- . benzamide; N-Cyclopropylmethoxy-3,4-diflucro-2-(4-iodo-phenylamino)-5- (pyridin-3-ylsulfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-5-{(3- = hydroxy-propyl)-pyridin-2-yimethyl-suifamoyl]-2-(4-iodo-2-methyl-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-2- yimethyl-sulfamoyi]-2-(4-iodo-2-methyl-phenylamino)-benzamide; 5-(benzyl- pyridin-2-ylmethyl-sulfamoyl)-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2- methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2- methyl-phenylamino)-5-{(pyridin-4-ylmethyl)-sulfamoyl]-benzamide; N- cyclopropylmethoxy-5-(ethyl-pyridin-4-ylmethyl-sulfamoyi)-3,4-difluoro-2-(4-iodo- 2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- 2-methyl-phenylamino)-5-(methyl-pyridin-4-yimethyl-sulfamoyl)-benzamide; N- cyclopropylmethoxy-3,4-difluoro-5-{(3-hydroxy-propyl)-pyridin-4-ylmethyl- sulfamoyl]-2-(4-iodo-2-methyl-phenylamino)-benzamide; N-cyclopropyimethoxy- 3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-4-ylimethyl-sulfamoyl]-2-(4-iodo-2-methyi-

phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl- phenylamino)-5-(methyl-phenyl-sulfamoyl)-benzamide; N-cyclopropylmethoxy- 3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-5-phenyisulfamoyl-benzamide; N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(pyridin-3- ylsulfamoyl)-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy- 3,4-difluoro-5-[(3-hydroxy-propyl)-pyridin-2-ylmethyl-sulfamoyl)-benzamide; 2-(2- chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy- ethyl)-pyridin-2-yimethyl-sulfamoyl]-benzamide; 5-(benzyl-pyridin-2-ylmethyi- sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- 5-[(pyridin-4-ylmethyl)-sulfamoyl]-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-5-(ethyl-pyridin-4-ylmethyl-sulfamoyl)-3,4-difluoro- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- 5-(methyl-pyridin-4-ylmethyl-sulfamoyl)-benzamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-[(3-hydroxy-propyl)-pyridin-4- yimethyl-sulfamoyl}-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-4-yimethyl- sulfamoyl]-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy- 3,4-difluoro-5-(methyl-phenyl-sulfamoyl)-benzamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-phenylsulfamoyl-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-(pyridin-3- ylsulfamoyl)-benzamide; N-cyclopropylmethoxy-2-(4-iodo-phenylamino)-4- phenylsulfamoyl-benzamide; N-cyclopropylmethoxy-2-(4-iodo-phenylamino)-4- (pyridin-3-ylsulfamoyl)-benzamide; N-cyclopropylmethoxy-2-(4-iodo- : phenylamino)-4-[(pyridin-3-yimethyl)-sulfamoyl]-benzamide; 4-(bis-pyridin-3- ylmethyl-sulfamoy!)-N-cyclopropyimethoxy-2-(4-iodo-phenylamino)-benzamide;

N-cyclopropylmethoxy-4-[(2-hydroxy-ethyl)-pyridin-4-ylmethyl-sulfamoyl}-2-(4- iodo-phenylamino)-benzamide; -cyclopropylmethoxy-2-(4-iodo-phenylamino)-4- (methyl-pyridin-3-ylmethyl-sulfamoyt)-benzamide; N-cyclopropylmethoxy-4-[(3- diethylamino-propyl)-pyridin-3-ylmethyl-sulfamoyi}-2-(4-iodo-phenylamino)- benzamide; N-cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4- phenylsulfamoyl-benzamide; N-cyclopropylmethoxy-2-(4-iodo-2-methyl-

phenylamino)-4-(pyridin-3-yisulfamoyl)-benzamide; N-cyclopropylmethoxy-2-(4- iodo-2-methyl-phenylamino)-4-[(pyridin-3-ylmethyl)-sulfamoyl}-benzamide; 4-(bis- pyridin-3-ylmethyl-sulfamoyl)-N-cyclopropylmethoxy-2-(4-iodo-2-methyl- phenylamino)-benzamide; N-cyclopropyimethoxy-4-[(2-hydroxy-ethyl)-pyridin-4- ylmethyl-sulfamoyl]-2-(4-iodo-2-methyl-phenylamino)-benzamide; N- cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4-(methyl-pyridin-3- ylmethyl-suifamoyl)-benzamide; N-cyclopropylmethoxy-4-[(3-diethylamino- propyl)-pyridin-3-ylmethyl-sulfamoyl]-2-(4-iodo-2-methyl-phenylamino)- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy-4- phenylsulfamoyl-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-4-(pyridin-3-ylsulfamoyl)-benzamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-4-[(pyridin-3-ylmethyl)-sulfamoyl]- benzamide; 4-(bis-pyridin-3-ylmethyl-sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-

N-cyclopropylmethoxy-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- «15 cyclopropylmethoxy-4-[(2-hydroxy-ethyl)-pyridin-4-ylmethyl-sulfamoyl]- ; benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-(methyl- . pyridin-3-yimethyl-sulfamoyl)-benzamide; and 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-4-[(3-diethylamino-propyl)-pyridin-3-ylmethyl-sulfamoyl]- benzamide.

Further examples include: PD 298469, 2-(2-Chloro-4-iodo-phenylamino)- 3,4-difluoro-N-methoxy-5-(4-methyl-piperazine-1-sulfonyl)-benzamide; PD 298470, 2-(2-Chloro-4-iodo-phenylamino)-5-[(2-diethylamino-ethyl)-methyl- sulfamoyl}-3,4-difluoro-N-methoxy-benzamide; PD 298450, 2-(2-Chloro-4-iodo- phenylamino)-3,4-difluoro-N-methoxy-5-(methyl-prop-2-ynyl-sulfamoyl)- benzamide; PD 298451, 1-[4-(2-Chloro-4-iodo-phenylamino)-2,3-difluoro-5- methoxycarbamoyl-benzenesulfonyl]-piperidine-3-carboxylic acid amide; PD 298452, 2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-methoxy-5-[methyl-(2- pyridin-2-yl-ethyl)-sulfamoyl}-benzamide; PD 298453, 2-(2-Chloro-4-iodo- phenylamino)-5-[(3-dimethylamino-propyl)-methyl-sulfamoyl]-3,4-difluoro-N- methoxy-benzamide; PD 298454, 2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-

N-methoxy-5-(4-pyridin-2-yl-piperazine-1-sulfonyl)-benzamide; PD 298455, 5- (Bis-(2-methoxy-ethyl)-sulfamoyl]-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-

methoxy-benzamide; PD 298456, 5-[Benzyl-(2-dimethylamino-ethyl)-sulfamoyl]-2- (2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-methoxy-benzamide; and PD 298457, N-Allyloxy-2-(2-chloro-4-iodo-phenylamino)-5-dimethyisuifamoyl-3,4- difluoro-benzamide; PD 298461, N-Allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4- difluoro-5-(methyl-prop-2-ynyl-sulfamoyi)-benzamide; PD 298462, N-Allyloxy-2- (2-chloro-4-iodo-phenylamino)-3,4-difluoro-5-[4-(4-fluoro-phenyl)-piperazine-1- sulfonyl]-benzamide; PD 298466, N-Allyloxy-5-[benzyl-(2-dimethylamino-ethyl)- sulfamoyi]-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzamide; PD 298468, 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-(4-methyl- piperazine-1-sulfonyl)-benzamide; and PD 298448, 2-(2-Chloro-4-iodo- phenylamino)-3,4-difluoro-5-(methoxy-methyl-sulfamoyl)-N-(2-morpholin-4-y|- ethoxy)-benzamide.

Particularly preferred compounds include: PD 298458, N-Allyloxy-2-(2- chloro-4-iodo-phenylamino)-3,4-difluoro-5-(4-methyl-piperazine-1-sulfonyl)- benzamide; PD 298459, N-Allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro- 5-(methyl-phenyl-sulfamoyl)-benzamide; PD 298460, 5-(Allyl-methyl-sulfamoyl)-

N-allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzamide; PD 298463, 1-[5-Allyloxycarbamoyl-4-(2-chloro-4-iodo-phenylamino)-2, 3-difluoro- benzenesulfonyl]-piperidine-3-carboxylic acid amide; PD 298464, N-Allyloxy-2-(2- chiloro-4-iodo-phenylamino)-5-[(3-dimethylamino-propyl)-methyl-sulfamoyi]-3,4- difluoro-benzamide; PD 298465, N-Allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4- difluoro-5-(4-pyridin-2-yl-piperazine-1-sulfonyl)-benzamide; and PD 298467, N-

Allyloxy-2-(2-chioro-4-iodo-phenylamino)-3,4-difluoro-5-(methoxy-methyil- sulfamoyl)-benzamide.

v

C. Synthesis

The disclosed compounds can be synthesized according to

Scheme 1 below. af A 2 EN §=0 Ry” “§=0 R2™ 5% woe

F Et,N F LDA/THF/-78 °C N + NHR,OR,——» -_— OL

Nod CH,Cl, CX Re Neg I

Re Re “OL Ry

One equivalent of appropriately substituted sulfonyl chloride is added to a : solution of one equivalent of appropriately substituted hydroxylamine and excess triethylamine in CH,Cl; or Et,O and stirred for 30 minutes. The triethylamine hydrochloride precipitate is separated by filtration and discarded. If necessary, the product is further purified by chromatography on silica column. The pure 2- ) fluor hydroxamic or hydroxamate product is then added to a solution of appropriately substituted lithium anilide prepared by adding LDA to the aniline in

THF at-78 °C. After stirring at room temperature for 16 hours, the reaction mixture is poured in to Et;O-HCL. Any precipitated solid is separated by filtration ) and discarded. The filtrate is concentrated and the resulting crude product is . purified on silica column to give the desired target product.

B The disclosed compounds can also be made by other synthetic organic methods, as well as automated or combinatorial methods.

The disclosed compounds can be synthesized according to the following two Schemes, or variants thereof (see also Example 1A).

Regarding the first step of synthetic Scheme 1A, the reaction of the aniline and the benzoic acid derivative generally is accomplished by mixing the benzoic acid with an equimolar quantity or excess of the aniline in an unreactive organic solvent such as tetrahydrofuran, or toluene, in the presence of a base such as lithium diisopropylamide, lithium hexamethyldisilazide, n-butyl lithium, sodium hydride, or sodium amide. The reaction generally is carried out at a temperature of about —78 °C to about 25 °C, and normally is complete within 2 hours to about 4 days. The product can be isolated by removing the solvent, for example by evaporation under reduced pressure, and further purified, if desired, by standard methods such as chromatography, crystallization, or distillation.

Turning to the second step, the 2-phenylamino benzoic acid derivative is next reacted with an equimolar quantity or excess of a nucleophile such as an aniline, a phenol, or a thiophenol by mixing in an unreactive organic solvent such as tetrahydrofuran, or toluene, in the presence of a base such as lithium diisopropylamide, lithium hexamethyldisilazide, n-butyl lithium, sodium hydride, or sodium amide. The reaction generally is carried out at a temperature of about — 78 °C to boiling, and normally is complete within 2 hours to about 4 days. The product can be isolated by removing the solvent, for example by evaporation under reduced pressure, and further purified, if desired, by standard methods such as chromatography, crystallization, or distillation.

Finally, regarding step 3, the 4-arylheteroatom-2-phenylamino benzoic acid derivative next is reacted with a nucleophile such as ammonia, an amine, an alcohol, hydrazine, a hydrazine derivative, or a hydroxylamine derivative in the presence of a peptide coupling reagent. Amines that can be employed include monomethylamine and aniline. Alcohols that can be employed include cyclobutylmethanol and phenol. Hydrazine derivatives that can be employed include N,N-dimethylhydrazine and 1-aminopiperidine. Hydroxylamine derivatives that can be employed include methoxylamine, N-ethyl-isopropoxy amine, and tetrahydrooxazine. Typical coupling reagents include 2-ethoxy-1-

L ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), 1,3-dicyclohexylcarbodiimide (DCC), bromo-tris-(pyrrolidino)-phosphonium hexafluorophosphate (PyBrOP) and (benzotriazolyloxy) tripyrrolidino phosphonium hexafluorophosphate (PyBOP).

The 4-arylheteroatom-2-phenylamino benzoic acid derivative and the nucleophile 5S normally are mixed in approximately equimolar quantities in an unreactive solvent such as dichloromethane, tetrahydrofuran, chloroform, or xylene, and an : equimolar quantity of the coupling reagent is added. A base such as triethylamine or diisopropylethylamine can be added to act as an acid scavenger if desired. The coupling reaction generally is complete after about 10 minutes to 2 hours, and the product is readily isolated by removing the reaction solvent, for instance by evaporation under reduced pressure, and purifying the product by standard methods such as chromatography or crystallizations from solvents such as acetone diethyl ether or ethanol.

Referring to synthetic Scheme 2A, an altermative method for making the compounds of the invention involves first coupling the benzoic acid derivative with ) the arylheteroatom nucleophile, and then reacting this 4-arylheteroatom benzoic acid derivative with an aniline. The final step involves the coupling of the 4-arylheteroatom-2-phenylamino benzoic acid derivative with the ammonia, amine, alcohol, hydrazine, hydrazine derivative, or hydroxylamine derivative with a peptide coupling reagent. The general reaction conditions for all of the steps in

B Scheme 2A are similar to those described above for synthetic Scheme 1A.

Scheme 1A

HOO R, HO 4 R,

R, jou . Sol Base Jone! + ArXH

R, “UR, R, ~ I R, RR,

L, L, X=N,0,S

R,

HO © R, RogNLO

Bs ho! R,-N-O-R, UN

R, I 7 I

R,~ X RRs R, RR,

AF RX

Ar

Bs Ry rr _______________]

Scheme 2A

HO_O

HO_O 4

Li Ry xl R, Base HN

Le aH AS

RY TR, Ry RX ) Ry I

L, } Af

X=N,0,S R,

Ry

HO__O

T H R, Ri~gN Ne) R,

Base N R, NOR H =D, me UR

Ry PRA! 7 R, RR I

Rr aX Rye X08

Ar

Rs R,

LJ

HO__O gy ©

OL

ON" Y °F 1 PD 195928 . HN APK IC50=30+8 nM

JC “1

Lo WO 01/05393 PCT/US00/18348

The disclosed compounds can be synthesized according to the following five schemes, or variants thereof. The abbreviation PyBOP is (benzotriazolyl- oxy)-tripyrrolidino phosphonium hexafluorophosphate. These synthetic strategies are further exemplified in Examples 1B-5B below:

Scheme 1B

HO.__O HO. .O R. A lo]

H CHa O H CH,

F HN N N

N TL LDA/THF R,ONHR, s PyBOP s

F Fog t F

H A Ph

PhSO2 H

HO.__O R N 0 y CH Hor© Lom 0 bh CHa “O (nBu)y,NHSO4 N RD! 0 [ Ji Oo 77° 52° sg’

F 3 F 2 | F 4

R;ONHR,

PyBOP

R._N_o

OY 4 CHa

N

0

To

F 5

Scheme 2B 5

HO.__O HO._.O hd CHa HOC ony yn CHa

F HN N N

+ LDATHE. KOH/MeOH x x= x

F SiCHa)s Fg Si(CHa), Fog (PhsP),PdCl, | =—Si(CH,),

HN rR. _N_o “0” H CH, t “CL

ASS

F 8

[}

Scheme 3B

NO, NO, NO,

NH, F ) NHNH,

H oO oO

HsCO.__O. OCH; Noy MN

A »

NO, NO, NO,

N N, N,

Ww 9a wy 9b Q 9¢

H,/RaNi | Haman H,/RaNi

NH, NH, NH, g 10a $ 10b 0 10¢

N N, N_ oO. _O HO. _.O

HO © H Rio H f Rio § Rio

N © , Op, NN

F = F = F J 11a 11b 11c

OR, | OR, | OR4 ] 1

R,N.__O LR R,N._O Ly Rue RoN._.O LR

N N N ol Ts Ce, TL.

F = F = F Pa 12a 12b 12¢

Scheme 4B 3-Aryl-1,2,5-thiadiazols

HQ_O _— HQ__O _—

N jot CL.

N

F X F Neg 12 13

X=0, NOH

HQ_O no Ch HAO H CH HAO H CH

N N

OO 00 Or

Fa " Fos Ns F 16 N-g

Y = solubilizing side chain

Scheme 5B 0 V0 0 v0 o

HO-¢ Ng Ng R,

NH H H H 0

F vo FF Ipa —_— —_—

R Re PyBOP Rs Rs Rs Rs NY

N

Re Re | Re lv 17 2

NQ NQ NH

3" or oh

H

/

F Si / N 7 av Oc Oe 18 19

The disclosed compounds can be synthesized according to the following four Schemes, or variants thereof. These synthetic strategies, which are suitable for conventional or combinatorial synthetic methods, are further exemplified in

Examples 1C-4C below.

{

Scheme 1C F od H,CO —~ H—, iPr2NEt H,CO -{) , 304 2A + =/ NH — = z pz oN

Cl-8§ F RoR R O F > po CH2CI2 rr 0 F wo LT aco

I. LDA, THF, -78 °C HO © R, 2.CO2, THF/EQ0 IF LHN 3.H* eo —( a ol + OL > ~ NS °F I

I Ra 0 F

H,CO =f

HO. __O

Ty b 7 \ rN ES

LiN(TMS)2 (2 moleq excess) noo —{ 0 | os | P _—_—————>» = N-§ F I

THF I Oo F co oo 1. "WH," coupling reagent, 1. "WH," coupling reagent, .~ solvent solvent oo 2. SOCI2 2TFA 7° td ~ pe w_O oT y Rs

Tonk Joust oe Noy c-s F 1

Jens oF

ANSE SP oF R7R6NH

I solvent \ wv _O

EE

N

= of I TL

RRN-S F I or

Scheme 2C

O w_.0O C1-S-OH w_O 7 H R, o H R

SNA WN

PR | or o

T R, 7 1 equivalent a-s R, l

R, O R w_O

Tm st solvent Ses R, 2 1

R’ 0 Rr

Scheme 3C

HO__O ~F RmewH ~F 1a F as” YR solvent NSTY UR 50m N-S R, oO R, R/ 6) R, 3H R, o R,

Rs

H

NA W_0

Li A HO. __O hd R, THER

UL Be 8 oy “WH 2A NA ! Ry. oll we ZN Rs. 0 | PZ St

LiIN(TMS)2 N-S YR, NCI coupling reagent, | N°S R, ! (2molegexcess) R’ © R, solvent R;, O R

Scheme 4C

HO__O

Th by 1)RONO, oo 0) DN ANE pdm orice (NN pp : Z EE — = ) 2) Reduction > 2) KMhO4 0:=S:0 0°8:0 0:8:0

OH OH OH

HOP W_0

NN w_0

Z - X 1) soc EN

I "“WH' —_— — [aN OQ, 0:S:0 0:=S:0 (2 moleq excess) Reagent, 5 N

Amine reagents such as RgR7NH in the schemes above are either commercially available or through straightforward madification of commercially available intermediates. Examples of such amine reagents, which can be reacted with the appropriate intermediate in a combinatorial or matrix method, are provided below.

S For example, in section B (Compounds), starting at page 8, line 16, there are three sets of thirty (one set each for Rs = H, Me, and Cl). The table below provides a number (corresponding to order that the name is found in the text; for example, “1” corresponds to compounds 1, 31, and 61 in the list of 90 compounds); the amine reagent name; and a Chemical Abstracts number.

Where a PD number is listed, the amine reagent was prepared from commercially available starting materials.

Number CAS # (position in | Amine reagent name or subset of 30 PD # 1] 3,3-dipicolylamine 1656-94-6 2 3,3-dipicolylamine 1656-94-6 3-(methylaminomethyl)pyridine 20173-04-0 3-(aminomethyl)pyridine 3731-52-0 “N-(3-diethylaminopropyl)-N-(pyridin-3-ylmethylamine” | PD 0096419 6] 3-(3-pyridyimethylamino)-1-propanal 6951-00-4 3-(ethylaminomethy|)pyridine PD 0133573 [8 | 2-(3-pyridylmethylamino)ethanol hydrochloride PD 0018185-0002 [9 | di-(2-picolyl)amine 1539-42-0 di-(2-picolyl)amine 1539-42-0 2-(methylaminomethyl)pyridine PD 0091430 2-(aminomethyl)pyridine 3731-519 3-(2-pyridylmethylamino)-1-propanol 6950-99-8 2-(2-pyridylmethylamino)ethano! PD 0018354 2-(N-benzylaminomethyl)pyridine PD 0054372 4-(aminomethyl)pyridine 3731-53-1 4-(methylaminomethyl)pyridine 3-(4-pyridyimethylamino)-1-propanol 7251-629 2-(4-pyridylmethylamino)ethanol hydrochloride PD 0018008-0002

N-methylaniline 100-61-8 22 | online 0] 62-53-3 462-08-8 62-53-3 462-08-8 3731-52-0 3,3-dipicolylamine 1656-94-6 2-(4-pyridylmethylamino)ethanol hydrochloride | PD 0018008-0002 3-(methylaminomethyl)pyridine 20173-04-0 “N-(3-diethylaminopropyl)-N-(pyridin-3-yimethyl)amine” | PD 0096419

Additional compounds within claim 1 can be made with the following amine reagents. The corresponding CAS number is provided. 5S 2-(methylamino)pyridine 4597-87-9 2-benzylaminopyridine 6935-27-9 2-allylaminopyridine 5866-28-4 2,2-dipyridylamine 1202-34-2 2-anilinopyridine 6631-37-4 2-aminopyridine 504-29-0 4-aminopyridine 504-24-5 2-benzylaminopyridine 6935-27-9 2-(4-methoxybenzyl)aminopyridine 52818-63-0 © 2-methylaminopyridine 4597-87-9

Combinatorial Synthesis

The following stock solutions were prepared: 1) An acetonitrile (anhydrous) stock solution 0.05 M in 5-chlorosulfonyl-2,3,4- trifluoro-benzoyl chloride. 2) Acetonitrile (anhydrous) stock solutions 0.05 M in each of the four appropriate hydroxylamine hydrochlorides (see list A) and 0.3 M in 2,6-lutidine. 3) Acetonitrile stock solutions 0.05 M in each of the 25 appropriate amines (see list B). Note that amine salts that were not soluble were also 0.1 Min 2,6- lutidine. 4) Acetonitrile (anhydrous) stock solutions in each of the 3 appropriate anilines (see list C) and 0.88 M in lithium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran).

An array which treated 4 hydroxylamine hydrochlorides independently with 5-chlorosulfonyl-2,3,4-trifluoro-benzoyl chloride, 25 amines, and 1 aniline was prepared to yield a total of 100 reactions. A liquid handling robot was used to transfer the reagents in such a manner as to insure that all possible combinations were achieved. The appropriate hydroxylamine hydrochloride solution (0.05 mmol, 1 mL) was added to a 2-dram vial, and each vial was treated with 5- chlorosulfonyl-2,3 4-trifluoro-benzoy! chloride solution (0.05 mmol, 1 mL). After

20 minutes the appropriate amine solution (0.05 mmol, 1 mL) was added sequentially. After a further 20 minutes the vials were treated with the solution of 4-iodoaniline (0.055 mmol, 1 mL). The vials were capped and shaken overnight at room temperature. The reactions were quenched with 1 mL of a 1 M aqueous ammonium chloride solution. The vials were concentrated to dryness under a stream of nitrogen and purified by reverse phase HPLC using a 30x100 mm YMC

ODS-A (C18) column. The mobile phase was acetonitrile/water (both with 0.05% trifluoroacetic acid) at 25 mL/min and a linear gradient of 10-100% over 6.5 min and then 3.5 min at 100%, detection was at 214 nm.

An array which treated 4 hydroxylamine hydrochlorides independently with 5-chlorosulfonyi-2,3,4-triflucro-benzoyl chloride, 25 amines, and 1 aniline was prepared to yield a total of 100 reactions. A liquid handling robot was used to transfer the reagents in such a manner as to insure that all possible combinations were achieved. The appropriate hydroxylamine hydrochloride solution (0.05 mmol, 1 mL) was added to a 2-dram vial, and each vial was treated with 5- chlorosulfonyl-2,3,4-trifluoro-benzoyl chloride solution (0.05 mmol, 1 mL). After + 20 minutes the appropriate amine solution (0.05 mmol, 1 mL) was added sequentially. After a further 20 minutes the vials were treated with the solution of « 4-iodo-2-methylaniline (0.05 mmol, 0.91 mL). The vials were capped and shaken overnight at room temperature. The reactions were quenched with 1 mL ofa 1 M aqueous ammonium chloride solution. The vials were concentrated to dryness under a stream of nitrogen and purified by reverse phase HPLC using a 30x100 mm YMC ODS-A (C18) column. The mobile phase was acetonitrile/water (both with 0.05% trifluoroacetic acid) at 25 mL/min and a linear gradient of 10-100% over 6.5 min and then 3.5 min at 100%, detection was at 214 nm.

An array which treated 4 hydroxylamine hydrochlorides independently with 5-chlorosulfonyl-2,3,4-triflucro-benzoyl chloride, 25 amines, and 1 aniline was prepared to yield a total of 100 reactions. A liquid handling robot was used to transfer the reagents in such a manner as to insure that all possible combinations were achieved. The appropriate hydroxylamine hydrochloride solution (0.05 mmol, 1 mL) was added to a 2-dram vial, and each vial was treated with 5- chlorosulfonyl-2,3,4-trifluoro-benzoyl chloride solution (0.05 mmol, 1 mL). After

20 minutes the appropriate amine solution (0.05 mmol, 1 mL) was added sequentially.

After a further 20 minutes the vials were treated with the solution of 2-chloro-4-iodoaniline (0.05 mmol, 0.91 mL). The vials were capped and shaken overnight at room temperature.

The reactions were quenched with 1 mL ofa 1 M aqueous ammonium chloride solution.

The vials were concentrated to dryness under a stream of nitrogen and purified by reverse phase HPLC using a 30x100 mm YMC ODS-A (C18) column.

The mobile phase was acetonitrile/water (both with 0.05% trifluoroacetic acid) at 25 mL/min and a linear gradient of 10-100% over 6.5 min and then 3.5 min at 100%, detection was at 214 nm.

Combinatorial Synthesis

Table of Example Reagents

List A-Hydroxylamines: 1. O-methyl-hydroxylamine 2. Oa-allyl-hydroxylamine hydrochloride monohydrate (Aldrich) 3. O-cyclopropylmethyl-hydroxylamine hydrochloride 4. O-(2-morpholin-4-yl-ethyl)-hydroxylamine hydrochloride

List B-Amines: 1. dimethylamine 2. diethylamine 3. isopropyl-methyl-amine 4. diisopropylamine 5. methylhydrazine 6. 1-methylpiperazine 7. N,N-diethyl-N’-methylethane-1,2-diamine 8. benzylmethylamine 9. dibenzylamine 10. methyl-phenyl-amine 11. allyl-methyl-amine 12.methyl-prop-2-ynyl-amine 13. methylamino-acetonitrile hydrochloride 14. 1-(4-fluoro-phenyl)-piperazine 15. furan-2-ylmethyl-methyl-amine 16. piperidine-3-carboxylic acid amide 17.methyl-phenethyl-amine 18. methyl-(2-pyridin-2-yl-ethyl)-amine 19.N,N,N-trimethyl-propane-1,3-diamine 20. methyl-(1-methyl-piperidin-4-yl)-amine 21.1-pyridin-2-yl-piperazine 22.bis-(2-methoxy-ethyl)-amine 23. N-benzyl-N,N-dimethyl-ethane-1,2-diamine 24. methylamino-acetic acid fert-butyl ester hydrochloride 25. O,N-dimethyl-hydroxylamine hydrochloride 40

List C-Anilines: 1. 4-iodoaniline 45 2. 2-chloro-4-iodoaniline 3. 4-iodo-2-methylaniline

« z = w / \. ff « 2

LL

= be 7° / ©” Yo

Low o / o of (T'S = A ° Yo / \

Ls °°

A o- Za

S 2 ple

T =z, Zz, aw = J, O T Ce & = 5 2 ww (VI wu [TI ° o uw {1 = 4 \ Oo } 2° Fo) = 3 o= \ A z— 2 oO A °

I QQ

0»

Chemical Examples

Example 1

Preparation of 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4- difluoro-benzenesulfonamide (PD 0297447)

N-cyclopropylmethoxy-2,3 4-trifluoro-benzenesulfonamide.

To a stirring suspension comprised of O-cyclopropylmethyl- hydroxylamine hydrochloride (5.40 g, 0.0437 mol) in dichloromethane (20 mi) at ambient temperature under a nitrogen atmosphere was added diisopropylethylamine (10.8 mi, 0.062 mol). A solution comprised of 2,3,4- trifluorobenzenesulfonyl chloride (Oakwood Products, Inc., 1.00 g, 4.34 x 10° mol) in dichloromethane (120 ml) was added dropwise to the reaction vessel - containing the stirring suspension over a 12 minute period. The reaction mixture was stirred for another 12 minutes and was quenched with 10 % aqueous hydrochloric acid (140 ml). The biphasic mixture was stirred vigorously for 16 hours. The layers were separated and the organic phase was dried (MgSO,) and concentrated to 6 ml volume. The concentrated solution was administered to a flash silica column (Biotage, 90 g of silica gel). ) Elution with dichloromethane afforded 0.8283 g of a white amorphous solid; 68 % yield; "H-NMR (400 MHz; CDCl, signal offset to & 7.03; values reported are uncorrected) 6 7.50 (m, 1H), 7.10 (s, 1H), 6.95 (m, 1H), 3.59 (d, 2H, J=7.2

Hz), 0.80 (m, 1H), 0.31 (m, 2H), 0.02 (m, 2H); "*F-NMR (376 MHz; CDCl) 3 — 122.65 (m, 1F), -129.37 (m, 1F), -156.20 (m, 1F); MS (APCI-) 280 (M-1, 100), 210 (55), 195 (45). : 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzenesulfonamide (PD 0297447).

To a stirring solution comprised of 2-chloro-4-iodoaniline in tetrahydrofuran (10 ml) at —78 °C under a nitrogen atmosphere was added a 1.0 M tetrahydrofuran solution of lithium bistrimethylsilylamide (6.2 ml, 6.2 x

102 mol) to form a green suspension. The suspension was stirred for five minutes before a stirring suspension comprised of lithiated N- cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide (prepared by adding 3.0 ml of the 1.0 M lithium bistrimethylsilylamide solution to a stirring solution comprised of N-cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide in 10 mi of tetrahydrofuran at —78 °C under nitrogen gas) was added via canula.

The cold bath was removed and the stirring suspension was stirred for one hour. The reaction mixture was quenched with 10 % aqueous hydrochloric acid (50 ml) and the biphasic mixture was concentrated in vacuo to an aqueous suspension that was extracted with diethyl ether (200 mi). The organic phase was dried (MgSO,) an d was concentrated in vacuo to afford a tan oil. The crude product was purified by flash chromatography. Elution with a gradient (hexanes-ethyl acetate 99:1 > (2 min) 9:1 > (25 min) 3:1 afforded 1.10 g of a white amorphous foam; 73 % yield; "H-NMR (400 MHz; DMSO) 5 7.69 (m, 1H), 7.59 (d, 1H, J=1.9 Hz), 7.34 (dd, 1H, J=8.7, 1.9 Hz), 7.27 (s, 1H), 7.00 (s, 1H), 6.95 (m, 1H), 6.43 (dd, 1H, J=8.7, 5.8 Hz), 3.52 (d, 2H,

J=7.5 Hz), 0.74 (m, 1H), 0.34 (m, 2H), 0.02 (m, 2H); 'SF-NMR (376 MHz;

CDCls) 6 —124.76 (m, 1F), -136.69 (d, 1F, J=18.3 Hz); MS (APCI+) 515 (M+1, 100); (APCI-) 513 (M-1, 50), 443 (73), 428 (100); IR (KBr) 1491 cm™; Anal.

Calcdffound for CisH14CIF2IN2O3S C, 37.34/36.54; H, 2.74/2.71; N, 5.44/5.15;

F, 7.38/7.57.

The APK [Cs for PD 0297447 is 0.965 uM.

EXAMPLE 1A

Preparation of 2,4-bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5-nitrobenzoic acid /

Step a: Preparation of 5-nitro-2,3 4-trifluorobenzoic acid

To gently stirring concentrated sulfuric acid (50 ml) was added fuming nitric acid (3.4 ml, 0.076 mol). Solid 2,3,4-trifluorobenzoic acid (10.00 g, 0.05565 mol) was added directly in increments. After stirring 45 minutes, the reaction mixture had become an orange homogeneous solution which was then poured over chilled water (400 ml). The resulting aqueous suspension was extracted with diethyl ether (3 x 200 ml). The combined extracts were dried with anhydrous magnesium sulfate and concentrated in vacuo to yield 12.30 g of a dull, light-yellow solid. Recrystallization from chloroform (50 ml) ‘ afforded 9.54 g of the pale yellow microcrystalline product; 78 % yield; m.p. ; ~~ TH-NMR (400 MHz; DMSO) & 14.29 (broad s, 1H), 8.43-8.38 (m, 1H); "*C- " NMR (100 MHz; DMSO) 3 162.41, 154.24 (dd, Jc.==270.1, 10.7 Hz), 148.35 (dd, Jc£=267.0, 9.2 Hz), 141.23 (dt, Jc.r=253.4 Hz), 133.95, 123.30 (d, Jc. ¢=2.2 Hz), 116.92 (dd, Jcr=18.2, 3.8 Hz); '*F-NMR (376 MHz; DMSO) 5 - = 120.50 to -120.63 (m), -131.133 to -131.27 (m), -153.63 to -153.74 (m).

Step b: Preparation of 2 4-bis-(2-chloro-4-iodo-phenylamino)-3-fiuoro-5- nitrobenzoic acid

To a stirring solution comprised of 2-chloro-4-iodoaniline (Lancaster, 98 %, 12.33 g, 0.04864 mol) in tetrahydrofuran (20 ml) at -78 °C under nitrogen was added a 2.0 M lithium diisopropylamide solution in tetrahydrofuran- heptane-ethylbenzene (Aldrich, 35 ml, 0.070 mol) with a syringe. The addition formed a thick suspension. After five minutes of stirring, a solution comprised of 5-nitro-2,3,4-trifluorobenzoic acid (5.00 g, 0.0226 mol) in tetrahydrofuran (30 ml) was added with a syringe to give a dark reaction mixture. The cold bath was removed and the reaction mixture stirred for 20 minutes. The cool reaction mixture was poured into ether (600 mi) containing an excess of hydrogen chloride. The red solution instantly turned to a yellow suspension as a precipitate formed. This precipitate was removed by vacuum filtration.

The filtrate was concentrated in vacuo to a red powder (10.5 g). The red powder was triturated with boiling chloroform (800 ml). The triturated solids 5S were collected by vacuum filtration to give an orange powder (2.42 g). The mother liquor from the trituration was concentrated in vacuo to give a red- orange solid (ca. 10 g undried). This solid was loaded onto a flash silica column. Elution with dichloromethane removed some impurities. Continuing elution with 1 % methanol in dichloromethane afforede ca. 4 g of a red solid.

This red solid was dissolved in hot absolute ethanol (100 ml). The solution was boiled down to 50 mi before dilution to 300 ml with hexanes. This solution was boiled to 150 ml and rediluted to 300 ml with hexanes to produce slight turbidity. The mixture was cooled in the refrigerator for three days, affording a yellow precipitate. The precipitate was collected by vacuum filtration and was dried with suction to afford 0.15 g of a yellow solid; 1 % yield; "H-NMR (400 MHz; DMSO) § 8.94 (s, 1H), 8.55 (s, 1H), 7.79 (d, 2H,

J=2.0 Hz), 7.61-7.57 (m, 2H), 6.90 (dd, 1H, J=8.5, 3.9 Hz), 6.84 (dd, 1H,

J=8.3, 6.6 Hz); "*F-NMR (376 MHz; DMSO) § -122.62 (s); MS (APCI+) 692 (6), 691 (8B), 690 (31), 689 (10), 688 (55), 171 (47), 130 (100); (APCI-) 691 (4), 690 (12), 689 (14), 688 (70), 687 (32), 686 (100), 506 (50), 453 (97); IR (KBr) 1523 cm’; Anal. calcd/found for: C1gH10CLFIoN;O4 C, 33.17/33.32; H, 1.47/1.73; N, 6.11/5.73; Cl, 10.31/10.04; F, 2.76/3.70; |, 36.89/34 .32.

The APK ICs for 2,4-bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5- nitrobenzoic acid is 29.6 nM.

EXAMPLE 1B 4-Fluoro-2-(4-methanesulfanyl-phenylamino)-benzoic acid (1).

To a solution of 4-(methylmercapto)aniline (3.1622 g, 0.02 mole) in

THF at ~78°C, a solution of LDA in THF (2M, 30 ml, 0.06 mole) was added and the reaction mixture stirred for 30 minutes at "78°C (Scheme 1E) . Solid 2,4- diflluoro benzoic acid (3.1622 g, 0.02 mole) was added and the reaction stirred for 16 hours while it wormed up to room temperature. The reaction mixture was pour in to ether saturated with HCI gas. HCI gas was bubbled into until precipitation of salts ceased. The precipitated salts were separated by filtration and discarded. The ether layer was concentrated to give 1 as a white solid. Yield 5.63 g (100%); mp 173-179 °C (DEC); "H-NMR (400 MHz;

CDCl3) 59.39 (s, 1H), 8.04 (dd, 1H, J=9.2, 6.8 Hz), 7.32-7.17 (AB quartet, . 4H), 6.74 (dd, 1H, J=12.1, 2.4 Hz), 6.46-6.41 (m, 1H), 2.51 (s, 3H); *C-NMR a (100 MHz; CDCl5) 8 172.79, 167.57 (d, Jc.r=253.4 Hz), 151.55 (d, Jcr=12.2

Hz), 136.83, 135.40 (d, Jc.r=12.2 Hz), 134.72, 128.31, 124.60, 106.51, 105.12 (d, Jo.F=22.9 Hz), 99.79 (d, Jer=26.7 Hz), 16.51; *F-NMR (376 MHz; CDCl,) §-101.39 to -101.46 (m); MS (APCI+) 278 (M+1, 100); IR (KBr) 3319, 1664,

Jr 1589, 1258 cm; Anal. calcdffound for: C14H1,FNO,S C, 60.64/60.99; H, 3 4.36/4.63; N, 5.05/4.80; S, 11.56/10.97.

EXAMPLE 2B 4-Fluoro-2-(4-methanesulfinyl-phenylamino)-benzoic acid (2).

A mixture of 1 (Scheme 1B) (0.286 g, 0.001031 mole) and oxaziridine (0.235 g, 0.0009 mole) in CHCI; (30 ml) at room temperature for 2 hours. The solvent was removed and the resulting brown oil chromatographed on silica column. Elution with CH,Cl, removed fast moving byproduct. Further elution with CH,Cl;:CH30H (9.5:05), R¢= 0.27, gave pure 2 as a light brown solid.

Yield 132.8 mg (50%); mp 191-192 °C; "H-NMR (400 MHz; CDCl5) § 9.77 (s, 1H), 8.08 (dd, 1H, J=8.9, 6.7 Hz), 7.70-7.39 (AB quartet, 4H), 6.98 (dd, 1H,

J=11.6, 2.4 Hz), 6.57-6.52 (m, 1H), 2.80 (s, 3H); "*C-NMR (100 MHz; CDCl5)

5170.76, 167.18 (d, Jc,=253.3 Hz), 149.33 (d, Jcr=12.2 Hz), 143.02, 139.50, 135.37 (d, Jc.,=12.2 Hz), 125.47, 122.32, 108.22, 106.35 (d, Jc.¢=22.8 Hz), 100.69, (d, Jc+=25.9 Hz), 43.75; MS (APCI+) 294 (M+1, 100); IR (KBr) 1673, 1592, 1228 cm’; Anal. calcd/ffound for: C14H1,FNO5S C, 57.33/57.48: H,

S 412/427; N, 478/467.

EXAMPLE 3B 4-Fluoro-2-(4-methanesulfonyl-phenylamino)-benzoic acid (3). } A solution of 1 (Scheme 1B) (0.4458 g, 0.00152 mole) and tetrabutyl- ammonium oxon (1.1 g, 0.0030 mole) in CHCl, (20 ml) was stirred at room temperature for 16 hours. TLC showed the presence of starting material; so additional 1.1 g (0.0030 mole) of the tetrabutylammonium oxon was added and reaction mixture stirred for 16 more hours. The reaction mixture was loaded on to a silica column and eluted with CH.Cl.:CHsOH (9.75:0.25) and the fast moving fraction collected and concentrated to give 3 as a white solid.

Yield, 0.3856 g (82%); mp 200-202 °C; "H-NMR (400 MHz; CDCls) 6 9.78 (s, 1H), 8.13 (dd, 1H, J=8.9, 6.5 Hz), 7.94-7.38 (AB quartet, 4H), 7.10 (dd, 1H,

J=11.3, 2.4 Hz), 6.66-6.61 (m, 1H), 3.09 (s, 3H); "*C-NMR (100 MHz; CDCl,) 8 171.52, 167.28 (d, Jc.r=254.9 Hz), 148.32, 145.21, 135.59 (d, Jc.r=11.5 Hz), 134.50, 129.39, 120.62, 108.74, 107.46 (d, Jcr=22.8 Hz), 101.61 (d, Jc. r=26.7 Hz), 44.78; F-NMR (376 MHz; CDCl3) § -100.29 to -100.45 (m); MS (APCI+) 310 (M+1, 100); (APCI-) 308 (M-1, 100); Anal. calcd/found for:

C14H12FNO4S0.75 H,0 C, 52.08/52.36; H, 4.22/3.88; N,4.34/4.26.

EXAMPLE 4B 2-methyl-4-trimethylsilanylethynyl-aniline (5)

To a solution of 4-iodo-2-methyl-aniline (2.33g, 10 mmol), bis(triphenylphosphine)palladium(ll)chloride (1.4g, 0.2 mmol), Cul (0.19 g, 0.1 mmol) in Et3N (40 ml) at ice-bath temperature, (trimethylsilyl)acetylene (1.18 g. 12 mmol) was added dropwise (Scheme 2B). After an hour stirring, the ice- bath was removed and the reaction mixture heated at 40°C (oil-bath temperature) for one hour; cooled to room temperature and the solvent removed. The residue was partitioned between H,O and Et;0. The Et,O layer was separated, dried (MgSO) and concentrated to give an oil. The oil was purified by silica column, eluting with CH,Cl,. The fraction with R; = 0.37 was collected and concentrated to give 2-methyl-4-trimethylsilanylethynyl- aniline as a dark brown oil.

Yield 1.50 g (83%).

EXAMPLE 5B 4-Fluoro-2-(2-methyl-4-trimethylsilanylethynyl-phenylamino)-benzoic acid (6)

Continuing after Example 4B, to a solution of 2-methyl-4-trimethyl- silanylethynyl aniline (1.50 g, 0.008 mole) in THF (10 ml) at 78°C, LDA (2 M in THF, 6 ml, 0.012 mole) was added and the mixture was stirred at “78°C for 30 minutes. Solid 2,4-difluoro-benzoic acid (0.633 g, 0.004 mole) was added and the stirred for 16 hours while it warmed up to room temperature. The solvents were removed and water (30 ml) and Et;O (50 ml) added to the oil wn residue. The mixture was stirred vigorously and the Et,O layer separated, . dried (MgS04) and concentrated to give a brown solid. The solid was purified on silica column, eluted with CH.Cl,. The fraction with R; = 0.37 was collected ie and concentrated to give a light brown solid. The solid was added to pentane; some insoluble brown particulate was separated by filtration and discarded. : The pentane layer was concentrated to give 6 as a light yellow solid. Yield 0.65 g (47%); mp 170-171°C; "H-NMR (400 MHz; CDCl,) § 9.33 (s, 1H), 8.05 (dd, 1H, J=8.9, 6.8 Hz), 7.43 (d, 1H, J=1.2 Hz), 7.35 (dd, 1H, J=8.2, 1.7 Hz), 7.25 (d, 1H, J=8.2 Hz), 6.53 (dd, 1H, J=11.8, 2.4 Hz), 6.47-6.42 (m, 1H), 2.25 (s, 3H), 0.26 (s, 9H); *C-NMR (100 MHz; CDCl) § 172.86, 167.61 (d, Jc.

F=253.3), 1561.24 (d, Jc.r=12.3 Hz), 138.28, 135.38 (d, Jcr=11.4 Hz), 134.85, 132.82, 130.63, 123.81, 119.91, 106.63, 105.23 (d, Jc.r=22.8 Hz), 104.77, 99.98 (d, Jc£=26.7 Hz), 94.05, 17.78, 0.00; MS (APCI+) 342 (M+1, 100); IR (KBr) 2151, 1661, 1249 cm; Anal. calcdfound for: CigH20FNO.Si C, 66.83/67.02; H, 5.90/6.00; N, 4.10/4.09; F, 5.56/5.45.

EXAMPLE 6B 4-Fluoro-2-(2-methyl-4-ethynyl-phenylamino)-benzoic acid (7).

To a solution of 6 in CH30H (30 ml), aqueous 1N KOH (10 ml) was added. After stirring at room temperature for 16 hours, the CH;OH was removed and the aqueous layer was acidified with 6N HCI (Scheme 2B). The resulting white precipitation was extracted in to Et,O, the Et,O layer was dried (MgSO) and concentrated to give 7 as tan colored solid. Yield 0.4274 g (91%); mp 177-178 °C; H-NMR (400 MHz; CDCl5) § 9.35 (s, 1H), 8.08-8.04 (m, 1H), 7.44 (s, 1H), 7.38-7.25 (m, 2H), 6.57 (d, 1H, J=11.8 Hz), 6.48-6.44 (m, 1H), 3.08 (s, 1H), 2.27 (s, 3H); C-NMR (100 MHz; CDCls) § 172.84, 167.61 (d, Jc+=253.3), 1561.15 (d, Jcr=12.3 Hz), 138.63, 135.40 (d, Jc.r=12.3

Hz), 135.00, 132.87, 130.81, 123.76, 118.79, 106.75, 105.33 (d, Jc¢=22.8

Hz), 100.03 (d, Jcr=26.0 Hz), 83.37, 17.83, 0.00; *F-NMR (376 MHz; CDCl,) 8 -101.24 to -101.31 (m); MS (APCI+) 270 (M+1, 100); IR (KBr) 3315, 1672, 1594, 1253 cm’; Anal. calcd/found for: Cy¢H12FNO, C, 71.37/71.08; H, 4.49/4.82; N, 5.20/5.09.

EXAMPLE 7B 1-(4-nitro-pheny)-1H-pyrrole (9a)

To a gently refluxing mixture of 4-nitroaniline (6.906 g, 0.05 mole), and sodium acetate (23 g, 0.28 mole) in acetic acid (100 ml) was added 2,5- dimethoxytetrahydrofuran (7.26 g, 7.12 ml, 0.055 mole) dropwise (Scheme 38). After refluxing for 3 hours, the reaction mixture was poured on to crushed ice (~250 ml), basified with 10 % sodium hydroxide (250 mi) and extracted with CHCl. The CHLCl, layer was dried (K2CO3) to afford the product as a dark brown oil. Yield 9.40 g (100 %).

EXAMPLE 8B 1-(4-nitro-phenyl)-1H-pyrazole (Sb)

A mixture of pyrrazole (6.808 g, 0.1 mole) tetrabutylammonium bromide (3.22 g, 0.01 male) and KOH (11.22 g, 0.2 mole) were ground together and sonicated for 16 hours. To this 1-fluoro-4-nitrobenzene (15.521 g, 11.67 mi,

‘ 0.11 mole) was added and the mixture sonicated for 24 hours. The reaction mixture was extracted with CH,Cl,. The CHCl, layer was dried (MgSO) and concentrated to give dark brown solid. This was purified by silica column chromatography. Elution with CH,Cl, (Rf = 0.44) gave the product as a light brown solid. Yield 8.80 g (47 %); mp 171-172 °C; Anal. caicd/found for:

CgH/N30, C, 57.14/56.52; H, 3.73/3.62; N, 22.21/21.95.

EXAMPLE 9B 3,5-dimethyl-1-(4-nitro-phenyl)-1H-pyrazole (9c)

To a solution of 4-nitro-phenyl-hydrazine (15.3 g, 0.1 mole) and 2,4- pentanedione (10.01 g, 10.27 ml, 0.1 mole) in EtOH (200 ml) were added 5 drops of concentrated HCI. The mixture was refluxed for 16 minutes; and the solvent removed to give a gummy product. This was purified by silica column chromatography. Elution with CH.CI. gave the desired product (R¢= 0.10) as a brown solid. Yield 7.22 g (33 %).

EXAMPLE 108 4-Pyrrol-1-yl-phenylamine (10a) i Catalytic reduction (H2/RaNi (5 g) /THF) of 1-(4-nitro-phenyf)- 1H- pyrrole (9.69 g, 0.05149 mole) at 51 psi gave crude product as an oil (Scheme x 3B). The product was purified by silica column chromatography. Elution with

CHCl; (Rf = 0.13) gave the pure product as white solid. Yield 8.06 g (99 %); mp 80-81 °C.

EXAMPLE 11B

In a manner similar to the preparation of 4-pyrrol-1-yl-phenylamine, the following were prepared: 4-1H-Pyrazol-1-yl-phenylamine (10b). Dark brown oil, yield 6.26 g (100 %).

Benzenamine, 4-(3,5-dimethyl-1H-pyrazzol-1-yl) (10c). Dark brown oil.

Yield 6.45 g (100 %).

EXAMPLE 12B 4-Fluoro-2-(4-pyrrol-1-yl-phenylamino)-benzoic acid (11a) - 5 To a solution of 4-pyrrol-1-yl-phenylamine (3.16 g, 0.02 mole) in THF (30 ml) at 778°C, a solution of LDA (2M, 15 ml, 0.03 mole) was added and the mixture stirred for 30 minutes. Solid 2 4-difluorobenzoic acid was added and the reaction mixture stirred for 16 hours as it warmed up to room temperature.

The solvent was removed and ether (100 ml) added to the dark oily residue.

This was stirred vigorously and the insoluble gummy precipitate separated by filtration. The gamy residue was dissolved in HzO, acidified to pH 1 with 10%

HCI, and extracted with Et;O. The Et,O layer was dried (MgSQ.,) and concentrated to give the target compound as a brown solid. Yield 2.74 g (93 %); mp 223-225 °C (DEC); *F-NMR (376 MHz; CDCl3) § -101.44 (s); MS (APCI+) 297 (M+1, 100); IR (KBr) 1658, 1526, 1254 cm. ) In a manner similar to the preparation of 4-Fluoro-2-(4-pyrrol-1-yl- phenylamino)-benzoic acid, the following were prepared: 4-Fluoro-2-(4-pyrazoi-1-yl-phenylamino)-benzoic acid (11b). Light brown solid, mp 212-213 °C. 2-[4-(3,5-Dimethyl-pyrazol-1-yi)-phenylamino]- 4-Fluoro benzoic acid (11c). Tan powder, mp 198 —200 °C.

EXAMPLE 1C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-5-dimethylsulfamoyl-3.4- difluoro-benzoic acid methyl ester (APK {C5p=222 nM)

Step a: Preparation of 1-dimethylsulfamoyl-2, 3 4-trifluorobenzene

To a gently stirring solution comprised of 2,3,4-trifluorobenzenesulfonyl chloride (5.70 g, 0.0247 mol) in 1,2-dichloroethane (200 ml) was introduced by bubbling gaseous anhydrous dimethylamine. The mixture became cloudy after several minutes and was subsequently washed with water (200 ml), 6 N aqueous hydrochloric acid (200 ml), brine (200 ml), was dried over anhydrous magnesium sulfate, and was concentrated in vacuo to obtain a yellow oil. The crude product was purified by flash chromatography. Elution with dichloromethane afforded 3.40 g of a white solid; 58 % yield; "H-NMR (400

MHz; CDCls) § 7.63-7.56 (m, 1H), 7.12-7.04 (m, 1H), 2.812 (s, 3H), 2.807 (s, 3H); "*F-NMR (376 MHz; CDCl) § =124.91 to —125.03 (m), —127.98 to — 128.03 (m), —156.41 to —156.53.

Step b: Preparation of 5-dimethylsulfamoyl-2.3 4-trifluoro-benzoic acid

To a cold (-78 °C) stirring solution comprised of 1-dimethylsulfamoyl- 2,3,4-trifluorobenzene in anhydrous tetrahydrofuran (60 ml) under a nitrogen - atmosphere was added a commercially available lithium diisopropylamide solution (Aldrich, 2.0 M in tetrahydrofuran/heptane/ethylbenzene, 7.5 mi, 0.0150 mol). After stirring for about ten minutes, the purple solution was transferred via canula to a cold, stirring, saturated carbon dioxide in diethyl ether solution (200 ml). The reaction mixture took on a dull burgundy color.

The cold bath was removed and the reaction mixture warmed to ambient temperature over one hour. The mixture was then carefully quenched with 10 % aqueous hydrochloric acid (200 ml). The layers were separated. The organic phase was extracted twice (200, 100 ml portions) with 10 % (wt.) aqueous sodium hydroxide. The combined aqueous alkaline extracts were treated with concentrated aqueous hydrochloric acid (100 mi) to pH 0. A white precipitate formed. The suspension was allowed to cool, then was extracted with diethyl ether (600 ml). The organic extract was dried over anhydrous magnesium sulfate and was concentrated in vacuo to afford 2.70 g of an off-white solid; 67.5 % yield; mp 225-228 °C; "H-NMR (400 MHz:

DMSO) & 14.08 (broad s, 1H}, 8.02-7.97 (m, 1H), 2.75 (s, 3H), 2.74 (s, 3H) '9F-NMR (376 MHz; DMSO) & -122.50 to 122.63 (m), 122.95 to —123.08 (m), -154.49 to -164.61 (m); MS (APCI+) 284 (M+1, 22), 238 (100); (APCI-) 282 (M-1, 85), 259 (94), 238 (46), 216 (91), 195 (100); IR (KBr) 1702 cm";

Anal. calcd/found for: CgHgF3NO4S C, 38.17/38.40; H, 2.85/2.90; N, 4.95/4.80;

F, 20.12/19.75; S, 11.3211.12.

Step c: Preparation of 5-dimethylsulfamoyl-2,3.4-trifluoro-benzoic acid methyl ssl The solid 5-dimethylsulfamoyl-2,3,4-trifluoro-benzoic acid (1.47 g, 0.00519 mol) and p-toluenesulfonic acid catalyst (17.1 mg) were dissolved in methanol (125 ml). The stirring mixture was brought to reflux under a nitrogen atmosphere for 51 hours. The reaction mixture was concentrated in vacuo to give a solid. The product was partitioned between diethyl ether (200 mi) and saturated aqueous potassium carbonate (75 ml). The layers were separated and the organic phase was washed with water (75 ml), brine (75 ml), was dried over anhydrous potassium carbonate, and was concentrated in vacuo to afford 0.15 g of an off-white solid; 10 % yield; "H-NMR (400 MHz; CDCl) § 8.23-8.19 (m, 1H), 3.92 (s, 3H), 2.83 (s, 6H); '*F-NMR (376 MHz; CDCl) & — 120.79 to —-121.02 (m), -153.69 to -153.80.

Step d: Preparation of 2-(2-chloro-4-iodo-phenylamino)-5-dimethylsulfamoy!- 3.4-difluoro-benzoic acid methyl ester

To a stirring cold (-78 °C) solution comprised of 2-chloro-4-iodoaniline (0.143 g, 5.64x10* mol) in anhydrous tetrahydrofuran (5 ml) under a nitrogen atmosphere was added a commercially available lithium diisopropylamide solution (Aldrich, 2.0 M in tetrahydrofuran/heptane/ethylbenzene, 0.300 mi, 6.0x10™ mol). After stirring for 5 minutes, a solution comprised of 5- dimethylsulfamoyl-2,3,4-trifluoro-benzoic acid methyl ester (0.15 g, 5.0x10™ mol) in tetrahydrofuran (10 ml) was added via syringe. The cold bath was removed and the reaction mixture was stirred for 2 hours. The reaction mixture was then partitioned between diethyl ether (125 ml) and saturated aqueous sodium bicarbonate (125 ml). The aqueous bicarbonate phase was : extracted with an additional portion (125 ml) of diethy! ether. The combined organic phases were dried over anhydrous magnesium sulfate and concentrated in vacuo to give a yellow oil. The oil was crystallized from heptane-ethyl acetate to afford 0.060 g of an off-white powder; 23 % yield; mp 154-156 °C; "H-NMR (400 MHz; CDCl3) § 9.74 (s, 1H), 8.30 (d, 1H, J=7.1 Hz), 7.72 (s, 1H), 7.49 (d, 1H, J=8.3 Hz), 6.73-6.69 (m, 1H), 3.92 (s, 3H), 2.84 (s, 3H), 2.83 (s, 3H); '"*F-NMR (376 MHz; CDCl) § -123.90 (d), =139.55 (d).

EXAMPLE 2C

Preparation _of 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4- difluoro-5-suifamoyl-benzamide (PD 219622)

Step a: Preparation of 1-bis-(4-methoxybenzyl)sulfamoyl-2.3 4- trifluorobenzene

To a stirring solution comprised of bis-4-methoxybenzylamine (2.5 g, 9.7x1 03 “ mol) and diisopropylethylamine (1.7 ml, 9.7x1 0°° mol) in dichloromethane (50 - ml) at 0 °C under nitrogen atmosphere was added liquid 2,3 ,4- trifluorobenzenesulfony! chloride (2.26 g, 9.5x10° mol) directly. The mixture was stirred cold for ten minutes. The ice-water bath was removed and the “ mixture was stirred for an additional 15 minutes and was then diluted with

E dichloromethane to 350 ml volume and was washed with saturated aqueous ammonium chloride (200 ml). The organic phase was dried (MgSQ,) and concentrated in vacuo to afford 4.99 g of a sticky white solid. The crude product was recrystallized from hexanes-acetone to afford 3.00 g of white needles; 70 % yield; mp 87-90 °C; "H-NMR (400 MHz; CDCl3) § 7.64-7.58 (m, 1H), 7.04-6.99 (m, 1H), [6.97 (d, 4H, J=8.5 Hz), 6.75 (d, 4H, J=8.8 Hz) AB q], 4.33 (s, 4H), 3.76 (s, 6H); '"F-NMR (376 MHz; CDCl;) 5 —125.44 to —125.56 (m),-128.61 to —128.72 (m), —156.91 to —157.03 (m); MS (APCI+) 121 (M- 330, 100); (APCI-) 330 (M-121, 18), 195 (M-256, 100); IR (KBr) 1612, 1517, 1508, 1465, 1258, 1240, 1156, 1037, 1030 cm™'; Anal. calcd/found for:

CxoH20F3NO4S C, 58.53/57.98; H, 4.47/4.61; N, 3.10/2.85.

Step b: Preparation of 5-bis-(4-methoxybenzyl)sulfamoyl-2,3,4-trifluorobenzoic acid

To a stirring solution comprised of 1-bis-(4-methoxybenzyl)sulfamoyi- 2,3,4-trifluorobenzene (2.95 g, 6.5x10° mol) in tetrahydrofuran (60 ml) at -78 °C was added a solution comprised of 2.0 M lithium diisopropylamide in tetrahydrofuran/heptane/ethylbenzene (Aldrich, 3.35 ml, 6.7x10°> mol). After several minutes of stirring, the dark solution was transferred via canula over five minutes to a stirring solution comprised of carbon dioxide (excess) in diethyl ether at —78 °C. A white precipitate immediately formed. The cold bath was removed and the reaction mixture was stirred at ambient temperature for 18 hours. The reaction mixture was quenched with 200 ml of dilute aqueous hydrochloric acid. The layers were separated and the organic phase was dried (MgSO.) and concentrated in vacuo to give 2.82 g of an off- white solid. Recrystallization from dichloromethane (150 ml) afforded 2.10g of the white powder product; 65 % yield; mp 158-161 °C; "H-NMR (400 MHz;

DMSO) & 7.80-7.76 (m, 1H), 7.05-6.74 (AB q, 8H, J=8.6 Hz), 4.33 (s, 4H), 3.66 (s, 6H); "*F-NMR (376 MHz; DMSO) 5 —123.28 to —123.36 (m), —124.12 to —124.21 (m), —-155.41 to —155.53 (m); MS (APCI-) 494 (M-1, 47), 216 (89), 195 (100); IR (KBr) 3420, 2954, 2838, 1695, 1613, 1512, 1347, 1238, 1152, 1079 cm’: Anal. calcdffound for: Ca3HaoFsNOsS C, 55.76/55.85: H, 4.07/4.02:

N, 2.83/2.71; F, 11.50/11.41; S, 6.47/6.25.

Step c: Preparation of 5-bis-(4-methoxybenzyl)sulfamoyl-2-(2-chloro-4- iodophenylamino)-3.4-diflucrobenzoic acid (PD 215729)

To a stirring solution comprised of 2-chloro-4-iodoaniline (0.53 g, 2.0x107 mol) in tetrahydrofuran (10 ml) at —78 °C under a nitrogen atmosphere was added a solution comprised of 1.0 M lithium bis(trimethylsilyl)amide in tetrahydrofuran (Aldrich, 4.1 ml, 4.1x10> mol).

Within several minutes the solution became a thick light-green suspension.

To this mixture was added a solution comprised of lithium 5-bis-(4- methoxybenzyl)sulfamoyl-2,3,4-trifluorobenzoate in tetrahydrofuran, which was prepared by adding 2.0 mi of the Aldrich lithium bis(trimethylsilyl}amide solution (0.0020 mmol) to a solution comprised of 5-bis-(4-

methoxybenzyl)sulfamoyl-2,3,4-trifluorobenzoic acid (1.00 g, 2.0x1 0° mol) in tetrahydrofuran (10 ml) at —=78 °C. The reaction mixture was stirred for 15 minutes and was then concentrated in vacuo to a crude semisolid. The semisolid was taken up into diethyl ether (250 ml) and was washed with 1 % aqueous hydrochloric acid (150 ml). The ether phase was then washed with neutral water (200ml, pH 4 after wash), a second portion of water (200 mi, pH 6 after wash), and brine (200 ml). The organic phase was then dried (MgSOQOy) and was concentrated in vacuo to give 1.88 g of a sticky residue which was crystallized from toluene-heptane to afford 1.12 g of an off-white powder; 76 % yield; mp 162-166 °C; "H-NMR (400 MHz; DMSO) § 9.86 (s, 1H), 7.92 (d, 1H, J=6.8 Hz), 7.86 (d, 1H, J=1.7 Hz), 7.60 (dd, 1H, J=8.5, 1.7 Hz), 7.06- 7.04/6.78-6.75 (AB q, 8H, J=8.5 Hz), 6.93-6.89 (m, 1H), 4.31 (s, 4H), 3.66 (s, 6H): '°F-NMR (376 MHz; DMSO) 5 —127.22 (d), —141.36 (d); MS (APCI+) 729 (M+1, 1), 256 (50), 121 (100); (APCI-) 727 (M-1, 100); IR (KBr) 1698, 1673, 1513, 1251 cm™; Anal. calcdffound for: CxgH24CIF2IN2O6S C, 47.78/47.93; H, 3.32/3.33; N, 3.84/3.80; Cl, 4.86/4.84; F, 5.21/5.46; 1, 17.41/17.16; S, : 4.40/4 29. ] Step d: Preparation of 5-bis-(4-methoxy benzyl)sulfamoyl-2-(2-chloro-4- iodophenylamino)-N-cyclopropylmethoxy-3 4-difluorobenzamide (PD 218774) s To a stirring solution comprised of 5-bis-(4-methoxybenzyl)sulfamoyl-2- (2-chloro-4-iodophenylamino)-3,4-difluorobenzoic acid (0.935 g, 1.28x1 03 mol), cyclopropylmethoxylamine hydrochloride (0.175 g, 1.42x107 mol), and diisopropylethylamine (0.75 ml, 4.26x10™ mol) in a 1:1 v/v tetrahydrofuran- dichloromethane mixture (50 ml) was added solid PyBOP ([benzotriazolyloxy]tripyrrolidino phosphonium hexafluorophosphate,

Advanced ChemTech, 0.76 g, 1.46x1 0° mol). The reaction mixture was stirred for one hour, was then evaporated to a crude residue which was purified by flash silica column chromatography. Elution with a gradient (25 % dichloromethane to 75 % dichloromethane in hexanes) afforded 0.63 g of the off-white powder product; 62 % yield; mp 70->300 °C; "H-NMR (400 MHz;

DMSO) § 11.92 (s, 1H), 9.35 (s, 1H), 7.60 (s, 1H), 7.50-7.45 (m, 1H), 7.34 (d, 1H, J=8.5 Hz), 6.82-6.54 (AB q, 8H, J=8.3 Hz), 6.59-6.54 (m, 1H), 4.09 (s,

4H), 3.46 (s, 6H), 0.90-0.80 (m, 1H), 0.30-0.25 (m, 2H), 0.03-0.00 (Mm, 2H); 'F-NMR (376 MHz; DMSO) § -129.05 (s), =140.23 (d, J=18.3 Hz); MS (APCI+) 798 (M+1, 70); (APCI-) 796 (M-1, 15), 726 (50), 131 (100); IR (KBr) 1642, 1611, 1584, 1513, 1478 cm"; Anal. calcd/found for: C33H31CIF2IN3O6S

C,49.67/49.88; H, 3.92/3.95; N, 5.27/5.19.

Step e: Preparation of 2-(2-chloro-4-iodophenylamino)-N-cyclopropylmethoxy- : 3,4-difluoro-5-sulfamoyl-benzamide (PD 219622)

A reaction solution comprised of 5-bis-(4-methoxybenzyl)sulfamoyl-2- (2-chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide (0.1010 g, 1.266x10™ mol) in trifluoroacetic acid (4 ml) was stirred at ambient temperature for 24 hours. The mixture was vacuum filtered and the precipitate rinsed with hexanes to afford 28.6 mg of a pale lavender powder; 42 % yield; mp 219-227 °C DEC; "H-NMR (400 MHz; DMSO) § 11.89 (s, 1H), 9.08 (s, 1H), 7.60 (s, 3H), 7.55 (d, 1H, J=6.9 Hz), 7.32 (d, 1H, J=8.6 Hz), 6.63- 6.59 (m, 1H), 3.40 (d, 2H, J=6.6 Hz), 0.90-0.80 (m, 1H), 0.30-0.26 (m, 2H), 0.05-0.00 (m, 2H); "*F-NMR (376 MHz; DMSO) 5 -130.61 (s), —140.38 (d, J=21.4 Hz); MS (APCI+) 558 (M+1, 70), 282 (100); (APCI-) 556 (M-1, 73), 486 (100); IR (KBr) 3390, 3283, 1652, 1513, 1477, 1163 cm’; Anal. calcdffound for: C17H15CIF2IN5O4S - 0.1 CoHF30; C, 36.30/36.31; H, 2.67/2.55; N, 7.38/7.00.

EXAMPLE 3C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-3 4-difluoro-N-hydroxy-5- sulfamoyl-benzamide (PD 224213)

To a stirring solution comprised of 5-bis-(4-methoxybenzyl)sulfamoyl-2- (2-chloro-4-iodophenylamino)-3,4-difluorobenzoic acid (0.67 g, 9.2x10™* mol),

O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (0.113 g, 9.65x10™ mol), and diisopropylethylamine (0.50 ml, 2.9x10™ mol) in a 1:1 v/v tetrahydrofuran- dichloromethane mixture (20 ml) was added solid PyBOP ([benzotriazolyloxyltripyrrolidino phosphonium hexafluorophosphate,

Advanced ChemTech, 0.52 g, 1.0x10 mol). The reaction mixture was stirred for 30 minutes, was concentrated in vacuo to a yellow oil, and was crystallized from methanol to afford 0.35 g of the off-white amorphous intermediate; 46 %

yield; the intermediate was dissolved in trifluoroacetic acid (10 ml) and was stirred at ambient temperature for 16 hours. The mixture was vacuum filtered to collect the precipitate, which was recrystallized from methanol-chloroform to afford 0.055 g of the tan powder product; 26 % yield from intermediate; mp 230-236 °C DEC; "H-NMR (400 MHz; DMSO) & 11.73 (s, 1H), 9.46 (s, 1H), 9.38 (s, 1H), 7.80-7.75 (m, 2H), 7.79 (s, 2H), 7.50 (d, 1H, J=8.5 Hz), 6.82-6.78 (m, 1H); '9F-NMR (376 MHz; DMSO) § ~130.83 (s), —139.24 (s); MS (APCI+) 504 (M+1, 53), 488 (90), 471 (100); (APCI-) 502 (M-1, 12), 486 (100); IR (KBr) 3295, 1652, 1636, 1519, 1477, 1315, 1157 cm’; Anal. calcd/found for: C43HgCIF2IN3O4S - 0.41 CHCI; C, 29.15/29.05; H, 1.72/1.66; N, 7.60/7.21.

EXAMPLE 4C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-3.4-difluoro-5-sutfamoyl- benzoic acid (PD 215730)

Solid 5-bis-(4-methoxybenzyl)sulfamoyl-2-(2-chloro-4- iodophenylamino)-3,4-difluorobenzoic acid (0.0995 g, 1.36x10™ mol) was ‘ dissolved in trifluoroacetic acid (5 ml) under a nitrogen atmosphere. The reaction mixture was stirred at ambient temperature for 65 hours. The mixture was vacuum filtered to isolate 55.2 mg of a fine white precipitate. The crude product was recystallized from chloroform to afford 31.8 mg of the fluffy white solid product; 48 % yield; mp 295-296 °C DEC; ‘H-NMR (400 MHz; DMSO) & 9.77 (s, 1H), 8.16 (d, 1H, J=7.3 Hz), 7.82 (s, 3H), 7.56 (d, 1H, J=8.5 Hz), 6.97- 6.92 (m, 1H); "*F-NMR (376 MHz, DMSO) § 128.47 (s), —141.13 (d, 19.8

Hz); MS (APCI+) 489 (M+1, 5), 102 (100); (APCI-) 491 (32), 490 (18), 489 (100), 488 (18), 487 (M-1, 75); IR (KBr) 3372, 3244, 1688 cm™; Anal. calcd/found for: C43HsCIF2IN,O4S C, 31.96/32.19; H, 1.65/1.81; N, 5.73/5.37.

EXAMPLE 5C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3.4- difluoro-5-dimethylsuifamoyl-benzamide (PD 250253)

Step a: Preparation of 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5- dimethylsulfamoyl-benzoic acid (PD 224339)

To a stirring solution comprised of 5-dimethylsulfamoyl-2,3,4- trifluorobenzoic acid (1.00 g, 3.53x10°® mol) in tetrahydrofuran (15 mi) at -78 °C under a nitrogen atmosphere was added a 1.0 M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (Aldrich, 3.6 ml, 3.6x107 mol). A lithium 2-chloro-4-iodoanilide suspension formed by adding a 1.0 M solution of lithium bis(trimethylsilyl)amide solution (7.2 ml, 7.2x1073 mol) to a solution comprised of 2-chloro-4-iodoaniline (0.94 g, 3.63x107 mol) in tetrahydrofuran (15 mi) at —78 °C was added via canula to the lithium 5-dimethylsulfamoyl- 2,3 4-trifluorobenzoate suspension. The cold bath was removed and the reaction mixture was stirred for one hour. The mixture was concentrated in vacuo to a crude solid. The crude product was suspended in diethyl ether (200 ml), to which suspension hydrogen chloride gas was introduced to produce a white precipitate. The precipitate was removed by vacuum filtration. The filtrate was concentrated in vacuo to give a dull-colored solid, which was triturated with hexanes-dichloromethane to afford 1.31 g of the white powder product; 72 % yield; mp 218-222 °C; "H-NMR (400 MHz;

DMSO) 6 9.89 (s, 1H), 8.06 (d, 1H, J=6.1 Hz), 7.85 (d, 1H, J=1.9 H2), 7.58 (dd, 1H, J=8.5, 1.9 Hz), 7.03 (dd, 1H, J=8.3, 6.6 Hz), 2.71 (s, 6H); "F-NMR (376 MHz; DMSO) 6 -125.58 (d, J=18.3 Hz), —140.14 (d, J=16.8 Hz); MS (APCI+) 519 (40), 518 (15), 517 (M+1, 100); (APCI-) 517 (6), 516 (2), 515 (M- 1, 5), 480 (45), 127 (100); IR (KBr) 3346, 1665, 1487, 1283 cm; Anal. calcdffound for: C1sH12CIF2IN2O4S C, 34.87/34.98; H, 2.34/2.32; N, 5.42/5.32.

Step b: Preparation of 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-5-dimethylsulfamoyl-benzamide

To a suspension comprised of 2-(2-chloro-4-iodo-phenylamino)-3,4- difluoro-5-dimethylsulfamoyl-benzoic acid (0.5 g, 9.68x10™ mol) and cyclopropylmethoxylamine hydrochloride (0.13 g, 1.05x103 mol) ina 1:1 viv mixture of dichloromethane-tetrahydrofuran (10 ml) was added diisopropylethylamine (0.65 ml, 3.73x10™ mol) followed by the addition of solid

PyBOP (0.55 g, 1.06x10™ mol). The reaction mixture was stirred at ambient temperature for three days. The mixture was concentrated in vacuo to a red oil. The crude product was treated with 10 % aqueous hydrochloric acid (150 mi) and was extracted with diethyl ether (150 ml). The organic phase was dried (MgSO.) and concentrated in vacuo to a crude solid. The solid was triturated with dichloromethane-hexanes and recovered by vacuum filtration to afford 0.3558 g of the white powder product; 63 % yield; mp 222-225 °C DEC; "H-NMR (400 MHz; DMSO) § 11.97 (s, 1H), 9.32 (s, 1H), 7.60 (d, 1H, J=1.9

Hz), 7.49 (d, 1H, J=5.8 Hz), 7.33 (dd, 1H, J=8.4, 1.9 Hz), 6.70 (dd, 1H, 8.4, 6.3 Hz), 3.43 (d, 2H, J=7.2 Hz), 2.53 (s, 6H), 0.87-0.83 (m, 1H), 0.30-0.25 (m, 2H), 0.03-0.00 (m, 2H); '°F-NMR (376 MHz, DMSO) § -127.67 (d, J=19.8 Hz), -139.32 (d, J=19.8 Hz); MS (APCI+) 586 (M+1, 100); (APCI-) 584 (M-1, 40), 514 (100); IR (KBr) 3263, 1644, 1585, 1507, 1480 cm’; Anal. calcd/found for:

C1oH1oCIF2IN30,4S C, 38.96/39.08; H, 3.27/3.18; N, 7.17/7 17.

EXAMPLE 6C

Preparation of N-cyclopropylmethoxy-3,4-difluoro-5-dimethylsulfamoyi-2-(4- . lodo-2-methyl-phenylamino)-benzamide (PD 252745) : Step a: Preparation of 3 4-difluoro-5-dimethylsulfamoyi-2-(4-iodo-2-methyl- : phenylamino)-benzoic acid (PD 224340)

Same procedure and same scale as Example 4C, Step a, except 4- iodo-2-methylaniline was used instead of 2-chloro-4-iodoaniline; afforded “0.9592 g of the off-white powder product; 55 % yield; mp 235-238 °C; "H-NMR (400 MHz; DMSO) & 9.69 (s, 1H), 8.04 (d, 1H, J=6.1 Hz), 7.60 (d, 1H, J=1.5

E Hz), 7.45 (dd, 1H, J=8.3, 1.7 Hz), 6.88 (dd, 1H, J=8.3, 5.4 Hz), 2.70 (s, 6H), 2.21 (s, 3H); "F-NMR (376 MHz; DMSO) § —126.25 (d, J=16.8 Hz), 142.74 (d, J=19.8 Hz); MS (APCI+) 497 (M+1, 69), 357 (70), 316 (100); (APCI-) 495 (M-1, 3), 127 (100); IR (KBr) 3240, 1686, 1512, 1473, 1341, 1151 cm”; Anal. calcd/found for: CigH1sF2IN204S C, 38.72/38.70; H, 3.05/3.01; N, 5.64/5.49.

Step b: Preparation of N-cyclopropyimethoxy-3,4-difluoro-5- dimethylsulfamoyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

Same procedure and same scale as Example 4C, Step b, except the product was purified by recrystallization from absolute ethanol to afford 0.1718 g of the pale yellow microcrystalline product; 28 % yield; mp 171-172 °C; H-

NMR (400 MHz; DMSO) § 11.79 (s, 1H), 8.91 (s, 1H), 7.40 (d, 1H, J=4.3 Hz),

7.36 (s, 1H), 7.21 (d, 1H, J=8.2 Hz), 6.54 (dd, 1H, 8.2, 4.3 Hz), 3.30 (d, 2H,

J=6.5 Hz), 2.52 (s, 6H), 2.00 (s, 3H), 0.85-0.75 (m, 1H), 0.29 (d, 2H, J=7.7

Hz), 0.01 (d, 2H, J=4.1 Hz); "*F-NMR (376 MHz; DMSO) 5 —128.94 (s), — - 143.32 (d, J=19.8 Hz), MS (APCI+) 566 (M+1, 100); (APCI-) 564 (M-1, 85), 494 (100); IR (KBr) 1649, 1609, 1588, 1512, 1475 cm; Anal. calcd/found for:

C20H22F2IN3O4S C, 42.49/42.42; H, 3.92/3.78; N, 7.43/7.40.

EXAMPLE 7C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-N-hydroxy-4- dimethylsulfamoyl-benzamide

Step a: Preparation of 4-methyl-benzene-N,N-dimethyisulfonamide

To a stirring solution comprised of para-toluenesulfonyl chloride in dichloromethane at 0 °C is introduced excess gaseous dimethylamine. The precipitate is removed by filtration and the filtrate is concentrated in vacuo to obtain the product.

Step b: Preparation of 4-methyl-3-nitro-benzene-N,N-dimethylsulfonamide

To a gently stirring solution comprised of 1 molar equivalent of fuming nitric acid in excess concentrated sulfuric acid is added 1 molar equivalent of 4-methyl-benzene-N,N-dimethylisulfonamide in increments. The mixture is stirred for one hour and then poured over chilled water. The mixture is extracted with a suitable solvent like diethyl ether or dichloromethane. The organic phase is dried over a suitable drying agent like magnesium sulfate and concentrated in vacuo to afford a crude product which may be purified by normal methods such as chromatography or crystallization from a solvent like chloroform or heptane.

Step c: Preparation of 3-amino-4-methyl-benzene-N.N-dimethylsulfonamide

The compound 4-methyl-3-nitro-benzene-N,N-dimethyisulfonamide is dissolved in ethanol. A catalyst like Raney nickel is added and the mixture hydrogenated in a shaker. The catalyst is removed by filtration. The solvent is removed in vacuo to give a product which may be purified if necessary by chromatography or crystallization from an appropriate solvent like chloroform or heptane-ethyl acetate.

Step d: Preparation of 3-fluoro-4-methyl-benzene-N,N-dimethylsulfonamide

The compound 3-amino-4-methyl-benzene-N,N-dimethylsulfonamide is diazotized with an alkyl nitrite like tert-butyl nitrite under anhydrous conditions in a non-reactive solvent like tetrahydrofuran or dichloromethane. The intermediate diazonium species is then treated with pyridinium fluoride to give the product, which may be purified by chromatography or crystallization.

Step e: Preparation of 4-dimethylsulfamoyl-2-fluoro-benzoic acid

A mixture comprised of 3-fluoro-4-methyl-benzene-N,N- dimethylsulfonamide and potassium permanganate (2.2 molar equivalents) in water is brought to reflux for four hours. The reaction mixture is filtered through celite. The filtrate is treated with activated carbon and refiltered through fresh celite. The second filtrate is acidified with concentrated hydrochloric acid to pH 0. The mixture is allowed to cool and is extracted with ia diethyl ether. The organic phase is dried over a drying agent like magnesium sulfate and is concentrated in vacuo. The product may be purified by recrystallization from an appropriate solvent like ethanol or chloroform. & Step f: Preparation of 2-(2-chloro-4-iodo-phenylamino)-4-dimethylsulfamoyl- : benzoic acid

To a stirring cold (-78 °C) solution comprised of 2-chloro-4-iodoaniline (1 molar equivalent) in anhydrous tetrahydrofuran under a nitrogen atmosphere is added a commercially available lithium diisopropylamide solution (Aldrich, 2.0 M in tetrahydrofuran/heptane/ethylbenzene, 1 molar equivalent). After stirring for 5 minutes, a solution comprised of 4- dimethylsulfamoy|-2-fluoro-benzoic acid (1 molar equivalent) in tetrahydrofuran is added. The cold bath is removed and the reaction mixture is stirred for 2 hours. The reaction mixture is then partitioned between diethyl ether and dilute aqueous hydrochloric acid. The organic phase is washed with brine, dried over magnesium sulfate, and concentrated in vacuo to afford a product which may be purified by chromatography of recrystallization from an appropriate solvent like chloroform or heptane-ethanol.

Step g: Preparation of 2-(2-chloro-4-iodo-phenylamino)-4-dimethylsulfamoyl- benzoic acid O-(tetrahydro-2H-pyran-2-yl)-oxyamide

A solution comprised of 2-(2-chloro-4-iodo-phenylamino)-4- dimethylsulfamoyl-benzoic acid, O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (1.25 molar equivalents), benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafiuorophosphate (1.25 molar equivalents), and diisopropylethylamine (3 molar equivalents) in 1:1 v/v tetrahydrofuran-dichloromethane is stirred for 30 minutes. The reaction mixture is concentrated in vacuo and the residue is purified by flash chromatography; elution with dichloromethane affords the desired product. The product may be recrystallized with an appropriate solvent like methanol if further purification is necessary.

Step h: Preparation of 2-(2-chloro-4-iodo-phenylamino)-4-dimethylsulfamoyl-

N-hydroxy-benzamide

The compound 2-(2-chloro-4-iodo-phenylamino)-4-dimethylsulfamoyi- benzoic acid O-(tetrahydro-2H-pyran-2-yl)-oxyamide is dissolved in an appropriate hydrogen chioride-saturated solvent like methanol or ethanol.

Once homogeneous, the solution is concentrated in vacuo to give the desired product. The product may be triturated with an appropriate solvent like chloroform or dichloromethane if further purification is necessary.

D. Uses

The disclosed compositions are useful as both prophylactic and therapeutic treatments for diseases or conditions relating to chronic pain, including neuropathic pain, as provided in the Summary section, as well as diseases or conditions modulated by the MEK cascade. For example, in one embodiment, the disclosed method relates to postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, crush injury, constriction injury, tissue injury, post-surgical pain, arthritis pain, or limb amputation

For example, local injuries can be treated with local or topical administration. Chronic pain affecting the entire body, such as diabetic neuropathy can be treated with systemic administration (injection or orally) of a disclosed composition. Treatment for chronic pain (e.g., post-operative

N pain) confined to the lower body can be administered centrally, e.g., : epidurally. Formulations and methods of administration can include the use of more than one MEK inhibitor, or a combination of a MEK inhibitor and another pharmaceutical agent, such as an anti-inflammatory, analgesic, muscle relaxing, or anti-infective agent. Preferred routes of administration are oral, intrathecal or epidural, subcutaneous, intravenous, intramuscular, and, for non-human mammals, intraplantar, and are preferably epidural. 1. Dosages

Those skilled in the art will be able to determine, according to known methods, the appropriate dosage for a patient, taking into account factors such as age, weight, general health, the type of pain requiring treatment, and the presence of other medications. In general, an effective amount will be between 0.1 and 1000 mg/kg per day, preferably between 1 and 300 mg/kg body weight, and daily dosages will be between 10 and 5000 mg for an adult subject of normal weight. Commercially available capsules or other formulations (such as liquids and film-coated tablets) of 100 mg, 200 mg, 300 mg, or 400 mg can be administered according to the disclosed methods. 2. Formulations

Dosage unit forms include tablets, capsules, pills, powders, granules, aqueous and nonaqueous oral solutions and suspensions, and parenteral solutions packaged in containers adapted for subdivision into individual doses.

Dosage unit forms can also be adapted for various methods of administration, including controlled release formulations, such as subcutaneous implants.

Administration methods include oral, rectal, parenteral (intravenous, intramuscular, subcutaneous), intracisternal, intravaginal, intraperitoneal, intravesical, local (drops, powders, ointments, gels, or cream), and by inhalation (a buccal or nasal spray).

Parenteral formulations include pharmaceutically acceptable aqueous or nonaqueous solutions, dispersion, suspensions, emulsions, and sterile powders for the preparation thereof. Examples of carriers include water, ethanol, polyols (propylene glycol, polyethylene glycol), vegetable oils, and injectable organic esters such as ethyl oleate. Fluidity can be maintained by the use of a coating such as lecithin, a surfactant, or maintaining appropriate particle size. Carriers for solid dosage forms include (a) fillers or extenders, (b) binders, (c) humectants, (d) disintegrating agents, (e) solution retarders, (f) absorption acccelerators, (g) adsorbants, (h) lubricants, (i) buffering agents, and (j) propellants.

Compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispensing agents; antimicrobial agents such as parabens, chlorobutanol, phenol, and sorbic acid; isotonic agents such as a sugar or sodium chloride; absorption-prolonging agents such as aluminum monostearate and gelatin; and absorption-enhancing agents. 3. Related compounds

The invention provides the disclosed compounds and closely related, pharmaceutically acceptable forms of the disclosed compounds, such as salts, esters, amides, hydrates or solvated forms thereof, masked or protected forms; and racemic mixtures, or enantiomerically or optically pure forms.

Pharmaceutically acceptable salts, esters, and amides include carboxylate salts (e.g., C 1g alkyl, cycloalkyl, aryl, heteroaryl, or non-aromatic heterocyclic), amino acid addition salts, esters, and amides which are within a reasonable benefit/risk ratio, pharmacologically effective, and suitable for. contact with the tissues of patients without undue toxicity, irritation, or allergic response. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactiobionate, and laurylsulfonate. These may include alkali metal and alkali earth cations such as sodium, potassium, calcium, and magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations such as tetramethyl ammonium, methylamine, trimethylamine, and ethylamine. See, v for example, S.M. Berge, et al., “Pharmaceutical Salts,” J. Pharm. Sci., 1977,

E 66:1-19 which is incorporated herein by reference. Representative pharmaceutically acceptable amides of the invention include those derived - from ammonia, primary C 1 alkyl amines and secondary di (C 1. alkyl) amines. Secondary amines include 5- or 6-membered heterocyclic or 5 heteroaromatic ring moieties containing at least one nitrogen atom and optionally between 1 and 2 additional heteroatoms. Preferred amides are derived from ammonia, C 1.3 alkyl primary amines, and di (C 4.2 alkyl)amines.

Representative pharmaceutically acceptable esters of the invention include

C17 alkyl, C 57 cycloalkyt, phenyl, and phenyl(C 1.)alkyl esters. Preferred esters include methyl esters.

The invention also includes disclosed compounds having one or more functional groups (e.g., hydroxyl, amino, or carboxyl) masked by a protecting group. Some of these masked or protected compounds are pharmaceutically acceptable; others will be useful as intermediates. Synthetic intermediates and processes disclosed herein, and minor modifications thereof, are also within the scope of the invention.

HYDROXYL PROTECTING GROUPS

Hydroxyl protecting groups include: ethers, esters, and protection for 1,2- and 1,3-diols. The ether protecting groups include: methyl, substituted methyl ethers, substituted ethyl ethers, substituted benzyl ethers, silyl ethers and conversion of silyl ethers to other functional groups.

Substituted Methyl Ethers

Substituted methyl ethers include: methoxymethyl, methylthiomethyl, t- utylthiomethyl, (phenyldimethylsilyl) methoxymethyl, benzyloxymethyl, p- ethoxybenzyloxymethyl, (4-methoxyphenoxy) methyl, guaiacolmethyl, t- butoxymethyl, 4-pentenyloxymethyl, siloxymethyl, 2-methoxyethoxymethyi, 2,2 2-trichloroethoxymethyl, bis(2-chloro- ethoxy)methyl, 2- (trimethylsilyl)ethoxymethyl, tetrahydropyranyl, 3-bromotetrahydro-pyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4- methoxytetrahydrothio-pyranyl, 4-methoxytetrahydrothiopyranyl S, S-dioxido, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, and 2,3,3a,4,5,6,7,7a-octahydro- 7,8,8-trimethyl-4,7-ethanobenzofuran-2-yl.

Substituted Ethyl Ethers

Substituted ethyl ethers include: 1-ethoxyethyl, 1-(2,chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2- fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilyethyl, 2-(phenylselenyl)ethyl, {- butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, and benzyl.

Substituted Benzyl Ethers

Substituted benzyl ethers include: p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyj, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl, p. pdinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, a-naphthyldiphenyl- methyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri-(p-methoxyphenyl)methyl, 4-(4’-bromophenacyloxy)phenyldiphenylmethyil, 4.,4' 4"tris(4,5-dichlorophthalimidophenyl)methyl, 4,4’,4"- tris(levulinoyloxyphenyl) methyl, 4, 4’ 4"tris(benzoyloxyphenyl)methyl, 3-

(imidazol-1-yimethyl)bis(4’,4"-dimethoxyphenyl)-methyl, 1,1-bis(4- methoxyphenyl)-1’'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl) xanthenyl, 9-(9- phenyl-10-oxo0) anthryl, 1,3-benzodithiolan-2-yl, and benzisothiazolyl S,S- dioxido.

Silyl Ethers

Silyl ethers include: trimethylsilyl, triethylsilyl, triisopropyisilyl, dimethylisopropyisilyl, diethylisopropylsilyl, dimethyithexyisilyl, butyldimethylsilyl, t-butyldiphenylsilyl, tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenyimethylsilyl, and t-butylmethoxyphenyisilyl.

ESTERS

Esters protecting groups include; esters, carbonates, assisted cleavage, miscellaneous esters, and sulfonates.

Esters «15 Examples of protective esters include: formate, benzoylformate, acetate, = chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate,

Ee methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p- chlorophenoxyacetate, p-P-phenylacetate, 3-phenylpropionate, 4- oxopentanoate (levulinate), 4,4-(ethylenedithio) pentanoate, pivaloate, : - 20 adamantoate,crotonate,4-methoxycrotonate, benzoate, p-phenylbenzoate, ‘ and 2,4,6-trimethylbenzoate (mesitoate).

Carbonates

Carbonates include: methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, : 2-(trimethylsilyl) ethyl, 2-(phenylsulfonyl) ethyl, 2-(triphenyiphosphonio) ethyl, isobutyl, vinyl, allyl, p-nitrophenyl, benzyl, p-methoxybenzyl, 3,4- dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, S-benzyl thiocarbonate, 4- ethoxy-1-naphthyl, and methyl dithiocarbonate.

Assisted Cleavage

Examples of assisted cleavage protecting groups include: 2-iodobenzoate, 4- azido-butyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl) benzoate, 2- formylbenzene-sulfonate, 2-(methylthiomethoxy) ethyl carbonate, 4-

(methylthiomethoxymethyl) benzoate, and 2-(methylthiomethoxymethyl) benzoate.

Miscellaneous Esters :

In addition to the above classes, miscellaneous esters include: 2,6-dichloro-4- methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl) phenoxyacetate, 2,4-bis(1,1-dimethylpropyl) phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2- butenoate (tigloate), o-(methoxycarbonyl) benzoate, p-P-benzoate, a- naphthoate, nitrate, alkyl N,N,N’ N '-tetramethylphosphorodiamidate, N- phenylcarbamate, borate, dimethylphosphinothioyl, and 2,4- dinitrophenylsulfenate.

Sulfonates

Protective sulfates includes: sulfate, methanesulfonate(mesylate), benzylsulfonate, and tosylate.

PROTECTION FOR 1,2- AND 1,3-DIOLS

The protection for 1,2 and 1,3-diols group includes: cyclic acetals and ketals, cyclic ortho esters, and silyl derivatives.

Cyclic Acetals and Ketals

Cyclic acetals and ketals include: methylene, ethylidene, 1-f-butylethylidene, 1-phenylethylidene, (4-methoxyphenyl) ethylidene, 2,2,2-trichloroethylidene, acetonide (isopropylidene), cyclopentylidene, cyclohexylidene, cycloheptylidene, benzylidene, p-methoxybenzylidene, 2,4- dimethoxybenzylidene, 3,4-dimethoxybenzylidene, and 2-nitrobenzylidene.

Cyclic Ortho Esters

Cyclic ortho esters include: methoxymethylene, ethoxymethylene, dimethoxy- methylene, 1-methoxyethylidene, 1-ethoxyethylidine, 1,2- dimethoxyethylidene, a-methoxybenzylidene, 1-(N,N- dimethylamino)ethylidene derivative, a-(N, N-dimethylamino) benzylidene derivative, and 2-oxacyclopentylidene.

PROTECTION FOR THE CARBOXYL GROUP

ESTERS

Ester protecting groups include: esters, substituted methyl esters, 2- substituted ethyl esters, substituted benzyl! esters, silyl esters, activated esters, miscellaneous derivatives, and stannyl esters.

Substituted Methyl Esters

Substituted methyl! esters include: 9-fluorenyimethyl, methoxymethyl, methyithiomethyl, tetrahydropyranyl, tetrahydrofuranyl, methoxyethoxymethyl, 2-(trimethylsilyl)ethoxy-methyl, benzyloxymethyl, phenacyl, p-bromophenacyl, o-methylphenacyl, p-methoxyphenacyl, carboxamidomethyl, and N- phthalimidomethyl. 2-Substituted Ethyl Esters 2-Substituted ethyl esters include: 2,2,2-trichloroethyl, 2-haloethyl, - chloroalkyl, 2-(trimethylsily)ethyl, 2-methylthioethyl, 1,3-dithianyl-2-methyl, 2(p-nitrophenylsulfenyl)-ethyl, 2-(p-toluenesulfonyl)ethyl, 2-(2'-pyridyl)ethyl, 2- " (diphenylphosphino)ethyl, 1-methyl-1-phenylethyl, t-butyl, cyclopentyl, cyclohexyl, allyl, 3-buten-1-yl, 4-(trimethylsily)-2-buten-1-yl, cinnamyl, a- - methylcinnamyl, phenyl, p-(methylmercapto)-phenyl, and benzyl.

Substituted Benzyl Esters = Substituted benzyl esters include: triphenylmethyl, diphenyimethyl, bis(o-nitrophenyl)methyl, 9-anthrylmethyl, 2-(9,10-dioxo)anthrylmethyl, 5- dibenzo-suberyl, 1-pyrenylmethyl,2-(trifluoromethyl)-6-chromylmethyl, 2,4,6- trimethylbenzyl, p-bromobenzyl, o-nitrobenzyl, p-nitrobenzyl, p- methoxybenzyl, 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl, 4-sulfobenzyl, piperonyl, and 4-P-benzyl.

Silyl Esters

Silyl esters include: trimethylsilyl, triethylsilyl, t-butyldimethyisilyl, i- propyldimethyisilyl, phenyldimethytsilyl, and di- t-butylmethylsilyl.

Miscellaneous Derivatives

Miscellaneous derivatives includes: oxazoles, 2-alkyl-1,3-oxazolines, 4-alkyl- 5-oxo-1,3-oxazolidines, 5-alkyl-4-oxo-1,3-dioxolanes, ortho esters, phenyl group, and pentaaminocobalt(lll) complex.

Stannyl Esters

Examples of stannyl esters include: triethylstannyl and tri-n-butylstannyl.

AMIDES AND HYDRAZIDES

Amides include: N,N —dimethyt, pyrrolidinyl, piperidinyt, 5,6- dihydrophenanthridinyl, o-nitroanilides, N-7-nitroindolyl, N-8-nitro-1,2,3,4- tetrahydroquinolyl, and p-P-benzenesulfonamides. Hydrazides include: N- phenyl, N,N '-diisopropyl and other dialkyl hydrazides.

PROTECTION FOR THE AMINO GROUP

CARBAMATES

Carbamates include: carbamates, substituted ethyl, assisted cleavage, : photolytic cleavage, urea-type derivatives, and miscellaneous carbamates.

Carbamates

Carbamates include: methyl and ethyl, 9-fluorenylmethyl, 9-(2- sulfo)fluorenylmethyl, 9-(2,7-dibromo)fluorenylmethyl, 2,7-di-t-butyi-[9-(10,10- dioxo-10,10,10,10-tetrahydro- thioxanthyl)Jmethyl, and 4-methoxyphenacyi.

Substituted Ethyl

Substituted ethyl protective groups include: 2,2,2-trichloroethyil, 2- trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl, 1,1-dimethyl- 2-haloethyl, 1,1dimethyl-2,2-dibromoethyl, 1,1-dimethyl-2,2,2-trichloroethyl, 1- methyl-1-(4-biphenylyl)ethyl, 1-(3,5-di-t-butylphenyl)-1-methylethyl, 2-(2’-and 4'-pyridyl)ethyl, 2-(N,N-icyclohexylcarboxamido)- ethyl, t-butyl, 1-adamantyl, vinyl, allyl, 1-isopropylallyl, connamyl, 4-nitrocinnamyl, quinolyl, N- hydroxypiperidinyl, alkyldithio, benzyl, p-methoxybenzyl, p-nitrobenzyl, p- bromobenzyl, p-chlorobenzyl, 2,4dichlorobenzyl, 4-methyisulfinylbenzyl, 9-anthrylmethyl, and diphenylmethyl.

C WO 01/05393 PCT/US00/18348

Assisted Cleavage

Protection via assisted cleavage includes: 2-methylthioethyl, 2-methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl, [2-(1,3-dithianyl)imethyl, 4-methylthiophenyl, 2,4-dimethyl-thiophenyl, 2-phosphoniocethyl, 2-triphenylphosphonioisopropyl, 1,1-dimethyl-2cyanoethyl, m-chloro-p- acyloxybenzyl, p-(dihydroxyboryl)benzyl, 5-benzisoxazolyl-methyl, and 2-(trifluoromethyl)-6-chromonylimethyil.

Photolytic Cleavage Photolytic cleavage methods use groups such as: m-nitrophenyl, 3,5- dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, and phenyl(o- nitrophenyl)methyl.

Urea-Type Derivatives

Examples of of urea-type derivatives include: phenothiazinyl-(10)-carbonyl derivative, N '-p-toluenesulfonylaminocarbonyl, and N ’- phenylaminothiocarbonyl. . Miscellaneous Carbamates

In addition to the above, miscellaneous carbamates include: t-amyl, S-benzyl thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl, » 20 cyclopropylmethyl, p-decyloxy-benzyl, diisopropylmethyl, 2,2- % dimethoxycarbonylvinyl, o-(N,N-dimethyl-carboxamido)-benzyl, 1,1-dimethyl- 3(N,N-dimethylcarboxamido)propyl, 1,1-dimethyl-propynyl, di(2-pyridy!)methyl, 2-furanylmethyl, 2-iodoethyl, isobornyl, isobutyl, isonicotinyl, p(p = methoxyphenyl- azo)benzyl, 1-methylcyclobutyl, 1-methylcyclohexyl, 1- methyi-1-cyclopropyl- methyl, 1-methyl-(3,5-dimethoxyphenyl)ethyl, 1-methyl- 1(p-henylazophenyl)- ethyl, 1-methyl-1-phenylethyl, 1-methyl-1-(4- pyridyl)ethyl, phenyl, p-(phenylazo)benzyl, 2,4,6-tri-t-butylphenyl, 4-(trimethylammonium) benzyl, and 2,4 ,6-trimethylbenzyl.

AMIDES

Amides

Amides includes: N-formyl, N-acetyl, N-chloroacetyl, N-trichloroacetyl,

N-trifluoroacetyl, N-phenylacetyl, N-3-phenylpropionyl, N-picolinoyl, N-3- pyridyl-carboxamide, N-benzoylphenylalanyl derivative, N-benzoyl, and N-p- phenylbenzoy!. :

Assisted Cleavage

Assisted cleavage groups include: N-o-nitrophenylacetyl, N-o- nitrophenoxyacetyl, N-acetoacetyl, (N-dithiobenzyloxycarbonylamino)acetyl, N-3-(p-hydroxphenyl) propionyl, N-3-(o-nitrophenyl)propionyl, N-2-methyl-2- (o-nitrophenoxy)propionyl, N-2-methyl-2-(o-phenylazophenoxy)propionyl, N-4- chlorobutyryl, N-3-methyl-3-nitrobutyryl, N-o-nitrocinnamoyl, N- acetylmethionine derivative, N-o-nitrobenzoyl, N-o- (benzoyloxymethyl)benzoyl, and 4,5-diphenyl-3-oxazolin-2-one.

Cyclic Imide Derivatives

Cyclic imide derivatives include: N-phthalimide, N-dithiasuccinoyl, - N-2,3-diphenyl-maleoyl, N-2,5-dimethylpyrrolyl,

N-1,1,4,4-tetramethyldisilylazacyclopentane adduct, 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl- 1,3,5-triazacyclohexan-2-one, and 1-substituted 3,5-dinitro-4-pyridonyl.

SPECIAL -NH PROTECTIVE GROUPS

Protective groups for — NH include: N-alkyl and N-aryl amines, imine derivatives, enamine derivatives, and N-hetero atom derivatives (such as N- metal, N-N, N-P, N-Si, and N-S), N-sulfenyl, and N-sulfonyl.

N-Alkyl and N-Aryl Amines

N-alkyl and N-aryl amines include: N-methyl, N-allyl,

N-[2-(trimethylsilyl)ethoxyl]-methyl, N-3-acetoxypropyl, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), quaternary ammonium salts,

N-benzyl, N-di(4-methoxyphenyl)methyl, N-5-dibenzosuberyl,

N-triphenyimethyl, N-(4-methoxyphenyl)diphenylmethyl, N-9-phenyifluorenyl,

N-2,7-dichloro-9-fluorenylmethylene, N-ferrocenyimethyl, and

N-2-picolylamine N '-oxide.

Imine Derivatives

Imine derivatives include: N-1,1-dimethylthiomethylene, N-benzylidene,

N-p-methoxybenzylidene, N-diphenyimethylene,

N-[(2-pyridyl)mesityljmethylene,

N-(N ',N '-dimethylaminomethylene), N,N -isopropylidene,

N-p-nitrobenzylidene,

N-salicylidene, N-5-chlorosalicylidene, N-(5-chloro-2-hydroxyphenyl)phenyl- methylene, and N-cyclohexylidene.

Enamine Derivative

An example of an enamine derivative is N- (5,5-dimethyl-3-oxo-1-cyclohexenyl).

N-Hetero Atom Derivatives N-metal derivatives include: N-borane derivatives, N-diphenylborinic acid derivative, N-[phenyl(pentacarbonylchromium- or -tungsten)]carbenyl, and

N-copper or N-zinc chelate. Examples of N-N derivatives include: N-nitro,

N-nitroso, and N-oxide. Examples of N-P derivatives include:

N-diphenylphosphinyl, N-dimethylthiophosphinyl, N-diphenylthiophosphinyl, N-dialkyl phosphoryl, N-dibenzy! phosphoryl, and N-diphenyl phosphoryl.

Examples of N-sulfenyl derivatives include: N-benzenesulfenyl,

N-o-nitrobenzenesulfenyl, N-2,4-dinitrobenzenesulfenyl,

N-pentachlorobenzenesulfenyl, N-2-nitro-4-methoxy-benzenesulfenyl, N- triphenylmethylsulfenyl, and N-3-nitropyridinesulfenyl. N-sulfonyl derivatives include: N-p-toluenesulfonyl, N-benzenesulfonyl, N-2,3,6-trimethyl- 4-methoxybenzenesulfonyl, N-2,4,6-trimethoxybenzenesulfonyl, N- 2,6-dimethyl-4-methoxy-benzenesulfonyl, N-pentamethylbenzenesulfonyl, N- 2,3,5,6-tetramethyl-4-methoxybenzene- sulfonyl, N- 4-methoxybenzenesulfonyl, N-2,4,6-trimethylbenzenesulfonyl, N- 2,6-dimethoxy- 4-methylbenzenesulfonyl, N- 2,2,5,7,8-pentamethylchroman-6-sulfonyl, N-methanesulfonyl,

N-p-trimethylsilylethanesulfonyl, N-9-anthracenesulfonyl, N- : 4-(4' 8'-dimethoxynaphthylmethyl)-benzenesulfonyl, N-benzylsulfonyl, N- trifluoromethyisulfonyl, and N-phenacylsuifony!.

Disclosed compounds which are masked or protected may be prodrugs, compounds metabolized or otherwise transformed in vivo to yield a disclosed compound, e.g., transiently during metabolism. This transformation may be a hydrolysis or oxidation which results from contact with a bodily fluid such as blood, or the action of acids, or liver, gastrointestinal, or other enzymes.

Features of the invention are further described in the examples below.

E. Examples

BIOLOGICAL EXAMPLES

Example 1

Effect of PD 198306 on streptozocin-induced static allodynia

Animals

Male Sprague Dawley rats (250-3009), obtained from Bantin and

Kingman, (Hull, U.K.) were housed in groups of 3. All animals were kept under a 12h light/dark cycle (lights on at 07h 00min) with food and water ad libitum.

All experiments were carried out by an observer blind to drug treatments.

Development of diabetes in the rat

Diabetes was induced in rats by a single i.p. injection of streptozocin (50 mg/kg) as described previously (Courteix et al., 1993).

Evaluation of static allodynia

Mechanical hypersensitivity was measured using Semmes-Weinstein von Frey hairs (Stoelting, Illinois, U.S.A.). Animals were placed into wire mesh bottom cages allowing access to the underside of their paws. Animals were habituated to this environment prior to the start of the experiment. Mechanical hypersensitivity was tested by touching the plantar surface of the animals right hind paw with von Frey hairs in ascending order of force ( 0.7, 1.2, 1.5, 2, 3.6, 5.5,8.5, 11.8, 15.1 and 29g) for up to 6 sec. Once a withdrawal response was established, the paw was re-tested, starting with the next descending von

Frey hair until no response occurred. The highest force of 29 g lifted the paw as well as eliciting a response, thus represented the cut off point. The lowest amount of force required to elicit a response was recorded as the paw withdrawal threshold (PWT) in grams.

Drugs

PD 198306 [N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl- phenylamino)-benzamide] and CI-1008 (pregabalin) were synthesized at

Parke-Davis (Ann Arbor, Mi, USA). PD 198306 was suspended in cremophor.ethanol:water (1:1:8) vehicle. Pregabalin was dissolved in water.

Both compounds were administered orally. Streptozocin (Aldrich, UK) was dissolved in 0.9% wiv NaCl and administered intraperitoneally. Drug administrations were made in a volume of 1 ml/kg.

Statistics

The static allodynia data were analysed using a Kruskail-Wallis ANOVA for non-parametric results, followed when significant by Mann-Whitney's t test.

Experimental protocol

Static allodynia was assessed with von Frey hairs, before (baseline,

BL) and 1h after oral administration of PD 198306 (30mg/kg, p.o.), vehicle (cremophor:ethanol:water, 1:1:8) or pregabalin (30mg/kg, p.o.) (test).

Animals were administered again the same compounds on the following day, both in the morning and the afternoon. Static allodynia was assessed only _ before and 1h after the afternoon administration, in order to minimise the habituation of the animals to the testing conditions. Animals treated with pregabalin received water in the morning administration, in order to avoid the potential development of tolerance to the compound with repeated administration.

Day 1: Day 2: a.m... PD 198306

Water

Vehicle p.m.: BL p.m.: BL

PD 198306 PD 198306

Pregabalin Pregabalin

Vehicle Vehicle

Test Test

RESULTS

A single administration of pregabalin (30mg/kg, p.o.) significantly blocked streptozocin-induced static allodynia 1h after administration. In contrast, a single administration of PD 198306 (30mg/kg, p.o) had no effect on streptozocin-induced static allodynia 1h after administration (see below).

However, after the compound had been administered twice more on the following day, it significantly blocked streptozocin-induced static allodynia 1h after the third administration. The effects had disappeared by the following day (see FIG. 1).

Example 2

MATERIALS AND METHODS

Animals

Male Sprague Dawley rats (250-3009), obtained from Charles River,

Margate, U.K.) were housed in groups of 3-6. All animals were kept under a 12h light/dark cycle (lights on at 07h 00min) with food and water ad libitum. All experiments were carried out by an observer blind to drug treatments.

Diabetes was induced in rats by a single i.p. injection of streptozocin (50mg/kg) as described previously (Courteix et al., 1993).

Development of Chronic Constriction Injury in the rat

Animals were anaesthetised with 2% isoflurane 1:4 O,/N>0 mixture maintained during surgery via a nose cone. The sciatic nerve was ligated as previously described by Bennett and Xie, 1988. Animals were placed on a homeothermic blanket for the duration of the procedure. After surgical preparation the common sciatic nerve was exposed at the middle of the thigh by blunt dissection through biceps femoris. Proximal to the sciatic trifurcation, about 7mm of nerve was freed of adhering tissue and 4 ligatures (4-0 silk) were tied loosely around it with about 1mm spacing. The incision was closed in layers and the wound treated with topical antibiotics. intrathecal injections

PD 1988306 and pregabalin were administered intrathecally in a volume of 10 pl using a 100 pl Hamilton syringe by exposing the spine of the rats under brief isoflurane anaesthesia. Injections were made into the intrathecal space between lumbar region 5-6 with a 10 mm long 27 gauge needle. Penetrations were judged successful if there was a tail flick response. The wound was sealed with an autoclip and rats appeared fully awake within 2-3 min following injection.

Evaluation of static allodynia

Mechanical hypersensitivity was measured using Semmes-Weinstein von Frey hairs (Stoelting, illinois, U.S.A). Animals were placed into wire mesh bottom cages allowing access to the underside of their paws. Animals were habituated to this environment prior to the start of the experiment. Mechanical hypersensitivity was tested by touching the plantar surface of the animais right hind paw with von Frey hairs in ascending order of force ( 0.7, 1.2, 1.5, 2, 3.6, 5.5, 8.5, 11.8, 15.1 and 29g) for up to 6sec. Once a withdrawal response was established, the paw was re-tested, starting with the next descending von

Frey hair until no response occurred. The highest force of 29g lifted the paw as well as eliciting a response, thus represented the cut off point. The lowest amount of force required to elicit a response was recorded as the paw withdrawal threshold (PWT) in grams.

Experimental protocol

Static allodynia was assessed with von Frey hairs, before (baseline,

BL) and 0.5h, 1h and 2h after intrathecal or intraplantar administration of PD 198306 (1-30pg, i.t.), vehicle (cremophor:ethanol:water, 1:1:8) or pregabalin (10ug, i.t). For oral administration experiments, static allodynia was assessed with von Frey hairs, before (baseline, BL) and 1h after oral administration of

PD 198306 (3-30mg/kg, p.o.), vehicle (cremophor:ethanol:water, 1:1:8) or pregabalin (30mg/kg, p.o.). Animals were administered again the same compounds on the following day, both in the morning and the afternoon. Static allodynia was assessed before and 1h after the morning administration. In the afternoon static allodynia was assessed before, 1h, 2h and 3h after administration for streptozocin treated animals. CCl animals were assessed before, 1h and 2h after administration

Drugs used

PD 198306 and pregabalin were synthesised at Parke-Davis (Ann

Arbor, MI, USA). PD 198306 was suspended in cremophor:ethanol:water (1:1:8) vehicle. Pregabalin was dissolved in water. Both compounds were administered orally, intrathecally or intraplantar in volumes of 1ml/kg, 10ul and 100ul respectively. Streptozocin (Aldrich, UK) was dissolved in 0.9% w/v NaCl and administered intraperitoneally in a volume of 1mikag.

Statistics

Data were analysed using a Kruskall-Wallis ANOVA for non-parametric results, followed when significant by Mann-Whitney's t test vs vehicle group.

RESULTS

1. Effects of PD 198306 on static allodynia, following systemic administration 1.1. Effect of PD198306 on streptozocin-induced static allodynia

A single administration of pregabalin (30mg/kg, p.o.) significantly blocked streptozocin-induced static allodynia 1h after administration. In contrast, a single administration of PD 198306 (3-30mg/kg, p.o) had no effect on streptozocin-induced static allodynia 1h after administration (FIG. 2).

However, after the compound had been administered twice more on the following day, PD 198306 (30mg/kg) significantly blocked streptozocin- induced static allodynia for 2h after the third administration (FIG. 2).

1.2. Effect of PD198306 on CCl-induced static allodynia

A single administration of pregabalin (30mg/kg, p.o.) significantly blocked CCI- induced static allodynia 1h after administration. In contrast, neither a single or multiple administration of PD 198306 (3-30mg/kg, p.o) had any effect on CCI- induced static allodynia (FIG. 3). 2. Effects of PD 198306 on static allodynia, following intrathecal administration

Intrathecally administered PD198306 (1-30ug) dose-dependently blocked the maintenance of static allodynia in both streptozocin (FIG. 4) and CCl animals (FIG. 5) with respective MEDs of 3 and 10 ug. This antiallodynic effect lasted for 1h. 3. Effects of PD 198306 on static allodynia, following intraplantar administration

An intrathecal administration of PD 198306 (30ug) significantly blocked static allodynia in both neuropathic pain models (FIGS. 6,7). In contrast, a single administration of PD 198306 at a dose 100-fold higher (3mg/100yl) directly into the paw had no effect on streptozocin (FIG. 6) or CCl-induced static allodynia (FIG. 7).

REFERENCES

Bennett GJ, Xie Y-K. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 1988,;33:87-107.

Courteix C, Eschalier A and Lavarenne J. Streptozocin —induced rats: behavioural evidence for a model of chronic pain. Pain 1993;53:81-8

Example 3

Effect of other MEK inhibitors in a neuropathic pain model in the rat

SUMMARY

The effect of several MEK inhibitors, with different binding affinities, has been investigated in the CCl mode! of neuropathic pain in the rat, by assessing static allodynia with von Frey hairs. Intrathecal administration of PD219622 or

PD297447 (30ug) had no significant effect on allodynia. This lack of effect may reflect the low affinity or solubility of the compounds. However, intrathecal administration of PD 254552 or PD 184352 (30ug), which posses higher binding affinities, blocked the maintenance of static allodynia in CCl animals. The antiallodynic effect was only evident for 30min post-injection and thus, shorter than the one observed for pregabalin (100ug). The magnitude of the effect was similar for 30ug of PD 184352 and 100ug of pregabalin. From this study it is concluded that MEK inhibitors exert an antiallodynic effect in

CCl-induced neuropathic rats when administered intrathecally, and that the antiallodynic effect correlates with the affinity of the compounds.

The animals and methods for developing chronic constriction injury in the rat, injecting test compounds, and evaluation of static allodynia were according to Example 2 above. PD219622, PD297447, PD 184352, PD 254552 and pregabalin were administered intrathecally at doses of 30ug for all

PD compounds and 100ug for pregabalin. Static allodynia was assessed with von Frey hairs, before (baseline, BL) and 0.5h, 1h and 2h after intrathecal administration of the compounds

Drugs used

PD297447, PD219622, PD 254552, PD 184352 (Cl-1040), and pregabalin were synthesised at Parke-Davis (Ann Arbor, MI, USA). PD297447,

PD219622, PD 254552 and PD 184352 were suspended in cremophor:ethanol:water (1:1:8) vehicle. Pregabalin was dissolved in water. All compounds were administered intrathecally in a 10pl volume.

Statistics

Data were analysed using a Kruskall-Wallis ANOVA for non-parametric results, followed when significant by Mann-Whitney's t test vs vehicle group.

RESULTS

Intrathecally administered PD297447 or PD219622 (30ug) had no significant effect on allodynia. This lack of effect may reflect the low affinity of the compounds (965nM and 100nM respectively). However, intrathecal administration of PD 184352 or PD 254552 (30ug) blocked the maintenance of static allodynia in CCl animals (see FIG. 8). These compounds possess higher affinity (2 and 5 nM respectively). The antiallodynic effect was only evident for 30min post-injection and thus, shorter than the one observed for pregabalin (100ug). The magnitude of the effect was similar for 30ug of PD 184352 and 100ug of pregabalin.

The results indicate that MEK inhibitors exert an antiallodynic effect in

CCl-induced neuropathic rats when administered intrathecally, and that the antiallodynic effect correlates with the affinity of the compounds. a3

CHEMICAL EXAMPLES

Example 1

Preparation of 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3.4- difluoro-benzenesulfonamide (PD 0297447)

N-cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide.

To a stirring suspension comprised of O-cyclopropylmethyl- hydroxylamine hydrochloride (5.40 g, 0.0437 mol) in dichloromethane (20 ml) at ambient temperature under a nitrogen atmosphere was added diisopropylethylamine (10.8 ml, 0.062 mol). A solution comprised of 2,3,4- trifluorobenzenesulfonyl chloride (Oakwood Products, Inc., 1.00 g, 4.34 x 103 mot) in dichloromethane (120 ml) was added dropwise to the reaction vessel containing the stirring suspension over a 12 minute period. The reaction mixture was stirred for another 12 minutes and was quenched with 10 % aqueous hydrochloric acid (140 ml). The biphasic mixture was stirred : vigorously for 16 hours. The layers were separated and the organic phase "was dried (MgS0,) and concentrated to 6 ml volume. The concentrated solution was administered to a flash silica column (Biotage, 90 g of silica gel). :

Elution with dichloromethane afforded 0.8283 g of a white amorphous solid, 68 % yield; "H-NMR (400 MHz; CDCl; signal offset to 5 7.03; values reported are uncorrected) 57.50 (m, 1H), 7.10 (s, 1H), 6.95 (m, 1H), 3.59 (d, 2H, J=7.2

Hz), 0.80 (m, 1H), 0.31 (m, 2H), 0.02 (m, 2H); '*F-NMR (376 MHz; CDCl;) & — 122.65 (m, 1F), -129.37 (m, 1F), -156.20 (m, 1F); MS (APCI-) 280 (M-1, 100), 210 (55), 195 (45). 2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzenesuffonamide (PD 0297447).

To a stirring solution comprised of 2-chloro-4-iodoaniline in tetrahydrofuran (10 ml) at =78 °C under a nitrogen atmosphere was added a 1.0 M tetrahydrofuran solution of lithium bistrimethyisilylamide (6.2 ml, 6.2 x 1073 mol) to form a green suspension. The suspension was stirred for five minutes before a stirring suspension comprised of lithiated N- cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide (prepared by adding 3.0 ml of the 1.0 M lithium bistrimethylsilylamide solution to a stirring solution comprised of N-cyclopropyimethoxy-2,3,4-trifluoro-benzenesulfonamide in 10 ml of tetrahydrofuran at -78 °C under nitrogen gas) was added via canula.

The cold bath was removed and the stirring suspension was stirred for one hour. The reaction mixture was quenched with 10 % aqueous hydrochloric acid (50 ml) and the biphasic mixture was concentrated in vacuo to an aqueous suspension that was extracted with diethyl ether (200 mi). The organic phase was dried (MgSQO,) an d was concentrated in vacuo to afford a tan oil. The crude product was purified by flash chromatography. Elution with a gradient (hexanes-ethyl acetate 99:1 > (2 min) 9:1 = (25 min) 3:1 afforded 1.10 g of a white amorphous foam; 73 % yield; "H-NMR (400 MHz; DMSO) § 7.69 (m, 1H), 7.59 (d, 1H, J=1.9 Hz), 7.34 (dd, 1H, J=8.7, 1.9 H2), 7.27 (s, 1H), 7.00 (s, 1H), 6.95 (m, 1H), 6.43 (dd, 1H, J=8.7, 5.8 Hz), 3.52 (d, 2H,

J=7.5 Hz), 0.74 (m, 1H), 0.34 (m, 2H), 0.02 (m, 2H); "*F-NMR (376 MHz; ~ CDCl3) 8 -124.76 (m, 1F), -136.69 (d, 1F, J=18.3 Hz); MS (APCI+) 515 (M+1, 100); (APCI-) 513 (M-1, 50), 443 (73), 428 (100); IR (KBr) 1491 cm; Anal.

Calcdffound for C1sH14CIF2IN2O3S C, 37.34/36.54; H, 2.74/2.71; N, 5.44/5.15;

F, 7.38/7.57.

The APK ICs for PD 0297447 is 0.965 pM.

EXAMPLE 1A

Preparation of 2.4-bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5-nitrobenzoic acid

Step a: Preparation of 5-nitro-2, 3 4-trifluorobenzoic acid

To gently stirring concentrated sulfuric acid (50 ml) was added fuming nitric acid (3.4 mi, 0.076 mol). Solid 2,3,4-trifluorobenzoic acid (10.00 g, 0.05565 mol) was added directly in increments. After stirring 45 minutes, the reaction mixture had become an orange homogeneous solution which was then poured over chilled water (400 ml). The resulting aqueous suspension was extracted with diethyl! ether (3 x 200 ml). The combined extracts were dried with anhydrous magnesium sulfate and concentrated in vacuo to yield 12.30 g of a dull, light-yellow solid. Recrystallization from chloroform (50 ml) afforded 9.54 g of the pale yellow microcrystalline product; 78 % yield; m.p. ; ) "H-NMR (400 MHz; DMSO) § 14.29 (broad s, 1H), 8.43-8.38 (m, 1H); °C-

NMR (100 MHz; DMSO) § 162.41, 154.24 (dd, Jc.7=270.1, 10.7 Hz), 148.35 (dd, Jc.¢=267.0, 9.2 Hz), 141.23 (dt, Jc¢=253.4 Hz), 133.95, 123.30 (d, Jc.

F=2.2 Hz), 116.92 (dd, Jc=18.2, 3.8 Hz); "F-NMR (376 MHz; DMSO) § - 120.50 to -120.63 (m), -131.133 to -131.27 (m), -153.63 to -153.74 (m).

Step b: Preparation of 2,4-bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5- nitrobenzoic acid

To a stirring solution comprised of 2-chloro-4-iodoaniline (Lancaster, 98 %, 12.33 g, 0.04864 mol) in tetrahydrofuran (20 mi) at -78 °C under nitrogen was added a 2.0 M lithium diisopropylamide solution in tetrahydrofuran- heptane-ethylbenzene (Aldrich, 35 ml, 0.070 mol) with a syringe. The addition formed a thick suspension. After five minutes of stirring, a solution comprised of 5-nitro-2,3,4-trifluorobenzoic acid (5.00 g, 0.0226 mol) in tetrahydrofuran (30 ml) was added with a syringe to give a dark reaction mixture. The cold bath was removed and the reaction mixture stirred for 20 minutes. The cool reaction mixture was poured into ether (600 ml) containing an excess of hydrogen chloride. The red solution instantly turned to a yellow suspension as a precipitate formed. This precipitate was removed by vacuum filtration.

The filtrate was concentrated in vacuo to a red powder (10.5 g). The red powder was triturated with boiling chloroform (800 ml). The triturated solids were collected by vacuum filtration to give an orange powder (2.42 g). The mother liquor from the trituration was concentrated in vacuo to give a red- orange solid (ca. 10 g undried). This solid was loaded onto a flash silica column. Elution with dichloromethane removed some impurities. Continuing elution with 1 % methanol in dichloromethane afforede ca. 4 g of a red solid. 10. This red solid was dissolved in hot absolute ethanol (100 ml). The solution was boiled down to 50 ml before dilution to 300 mi with hexanes. This solution was boiled to 150 ml and rediluted to 300 ml with hexanes to produce slight turbidity. The mixture was cooled in the refrigerator for three days, : affording a yellow precipitate. The precipitate was collected by vacuum filtration and was dried with suction to afford 0.15 g of a yellow solid; 1 % yield; "H-NMR (400 MHz; DMSO) 5 8.94 (s, 1H), 8.55 (s, 1H), 7.79 (d, 2H,

J=2.0 Hz), 7.61-7.57 (m, 2H), 6.90 (dd, 1H, J=8.5, 3.9 Hz), 6.84 (dd, 1H,

J=8.3, 6.6 Hz); '°F-NMR (376 MHz; DMSO) § -122.62 (s); MS (APCI+) 692 (6), 691 (8), 690 (31), 689 (10), 688 (55), 171 (47), 130 (100); (APCI-) 691 (4), 690 (12), 689 (14), 688 (70), 687 (32), 686 (100), 506 (50), 453 (97); IR (KBr) 1523 em’; Anal. calcdffound for: CygH1oClFI2N304 C, 33.17/33.32; H, 1.47/1.73; N, 6.11/5.73; CI, 10.31/10.04; F, 2.76/3.70; |, 36.89/34.32.

The APK ICs, for 2,4-bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5- nitrobenzoic acid is 29.6 nM.

. EXAMPLE 1B 4-Fluoro-2-(4-methanesulfanyl-phenylamino)-benzoic acid (1).

To a solution of 4-(methylmercapto)aniline (3.1622 g, 0.02 mole) in

THF at “78°C, a solution of LDA in THF (2M, 30 ml, 0.06 mole) was added and the reaction mixture stirred for 30 minutes at “78°C (Scheme 1E) . Solid 2,4- diflluoro benzoic acid (3.1622 g, 0.02 mole) was added and the reaction stirred for 16 hours while it wormed up to room temperature. The reaction mixture was pour in to ether saturated with HCI gas. HCI gas was bubbled into until precipitation of salts ceased. The precipitated salts were separated by filtration and discarded. The ether layer was concentrated to give 1 as a white solid. Yield 5.63 g (100%); mp 173-179 °C (DEC); ‘H-NMR (400 MHz;

CDCl) 5 9.39 (s, 1H), 8.04 (dd, 1H, J=9.2, 6.8 Hz), 7.32-7.17 (AB quartet, 4H), 6.74 (dd, 1H, J=12.1, 2.4 Hz), 6.46-6.41 (m, 1H), 2.51 (s, 3H); ’C-NMR ; (100 MHz; CDCl3) 8 172.79, 167.57 (d, Jc.r=253.4 Hz), 151.55 (d, Jor=12.2 ) Hz), 136.83, 135.40 (d, Jc.,=12.2 Hz), 134.72, 128.31, 124.60, 106.51, 105.12 (d, Jer=22.9 Hz), 99.79 (d, Jc£=26.7 Hz), 16.51; *F-NMR (376 MHz; CDCl5) 8 -101.39 to -101.46 (m); MS (APCI+) 278 (M+1, 100); IR (KBr) 3319, 1664, 1589, 1258 cm’; Anal. calcdffound for: C14H12,FNO,S C, 60.64/60.99; H, : 4.36/4.63; N, 5.05/4.80; S, 11.56/10.97.

EXAMPLE 2B 4-Fluoro-2-(4-methanesulfinyl-phenylamino)-benzoic acid (2).

A mixture of 1 (Scheme 1B) (0.286 g, 0.001031 mole) and oxaziridine (0.235 g, 0.0009 mole) in CHCI3 (30 mi) at room temperature for 2 hours. The solvent was removed and the resulting brown oil chromatographed on silica column. Elution with CHCl; removed fast moving byproduct. Further elution with CH2Cl:CH3OH (9.5:05), Rf = 0.27, gave pure 2 as a light brown solid. © 30 Yield 132.8 mg (50%); mp 191-192 °C; "H-NMR (400 MHz; CDCl3) 5 9.77 (s, 1H), 8.08 (dd, 1H, J=8.9, 6.7 Hz), 7.70-7.39 (AB quartet, 4H), 6.98 (dd, 1H,

J=11.6, 2.4 Hz), 6.57-6.52 (m, 1H), 2.80 (s, 3H); *C-NMR (100 MHz; CDCl;) 5 170.76, 167.18 (d, Jc-r=253.3 Hz), 149.33 (d, Jc.r=12.2 Hz), 143.02, 139.50,

135.37 (d, Jcr=12.2 Hz), 125.47, 122.32, 108.22, 108.35 (d, Jc.r=22.8 Hz), 100.69, (d, Jcr=25.9 Hz), 43.75; MS (APCI+) 294 (M+1, 100); IR (KBr) 1673, 1592, 1228 cm; Anal. calcd/found for: C14H12FNO3S C, 57.33/57.48; H, 4.12/4.27, N, 4.78/4.67.

EXAMPLE 3B 4-Fluoro-2-(4-methanesulfonyl-phenylamino)-benzoic acid (3).

A solution of 1 (Scheme 1B) (0.4458 g, 0.00152 mole) and tetrabutyi- ammonium oxon (1.1 g, 0.0030 mole) in CH,C!; (20 ml) was stirred at room temperature for 16 hours. TLC showed the presence of starting material; so additional 1.1 g (0.0030 mole) of the tetrabutylammonium oxon was added and reaction mixture stirred for 16 more hours. The reaction mixture was loaded on to a silica column and eluted with CH,Cl,:CH;0H (9.75:0.25) and the fast moving fraction collected and concentrated to give 3 as a white solid.

Yield, 0.3856 g (82%); mp 200-202 °C; ‘H-NMR (400 MHz; CDCl) § 9.78 (s, 1H), 8.13 (dd, 1H, J=8.9, 6.5 Hz), 7.94-7.38 (AB quartet, 4H), 7.10 (dd, 1H,

J=11.3, 2.4 Hz), 6.66-6.61 (m, 1H), 3.09 (s, 3H); >*C-NMR (100 MHz; CDCls) 8171.52, 167.28 (d, Jcr=254.9 Hz), 148.32, 145.21, 135.59 (d, Jc.r=11.5 Hz), 134.50, 129.39, 120.62, 108.74, 107.46 (d, Jcr=22.8 Hz), 101.61 (d, Jc. ¢=26.7 Hz), 44.78; 'F-NMR (376 MHz; CDCl) 8 -100.29 to -100.45 (m); MS (APCI+) 310 (M+1, 100); (APCI-) 308 (M-1, 100); Anal. calcd/found for:

C14H12FNO4S0.75 HO C, 52.08/52.36; H, 4.22/3.88; N,4.34/4.26.

EXAMPLE 4B 2-methyl-4-trimethylsilanylethynyl-aniline (5)

To a solution of 4-iodo-2-methyl-aniline (2.33g, 10 mmol), bis(triphenyiphosphine)palladium(il)chioride (1.4g, 0.2 mmol), Cut (0.19 g, 0.1 mmol) in EtsN (40 ml) at ice-bath temperature, (trimethylsilyl)acetylene (1.18 g, 12 mmol) was added dropwise (Scheme 2B). After an hour stirring, the ice- bath was removed and the reaction mixture heated at 40°C (oil-bath temperature) for one hour; cooled to room temperature and the solvent removed. The residue was partitioned between H,O and Et,O. The Et,O layer was separated, dried (MgSO,) and concentrated to give an oil. The oil was purified by silica column, eluting with CH,Cl,. The fraction with R= 0.37 was collected and concentrated to give 2-methyl-4-trimethylsilanylethynyl- aniline as a dark brown oil.

Yield 1.50 g (83%).

EXAMPLE 5B 4-Fluoro-2-(2-methyl-4-trimethylsilanylethynyl-phenylamino)-benzoic acid (6)

Continuing after Example 4B, to a solution of 2-methyl-4-trimethyl- silanylethynyl aniline (1.50 g, 0.008 mole) in THF (10 mi) at 778°C, LDA (2 M in THF, 6 ml, 0.012 mole) was added and the mixture was stirred at ~78°C for 30 minutes. Solid 2,4-difluoro-benzoic acid (0.633 g, 0.004 mole) was added and the stirred for 16 hours while it warmed up to room temperature. The solvents were removed and water (30 ml) and Et;O (50 ml) added to the oil residue. The mixture was stirred vigorously and the Et,O layer separated, dried (MgS0O,) and concentrated to give a brown solid. The solid was purified on silica column, eluted with CHCl. The fraction with Rs = 0.37 was collected and concentrated to give a light brown solid. The solid was added to pentane; some insoluble brown particulate was separated by filtration and discarded.

The pentane layer was concentrated to give 6 as a light yellow solid. Yield 0.65 g (47%); mp 170-171°C; "H-NMR (400 MHz; CDCl3) 5 9.33 (s, 1H), 8.05 (dd, 1H, J=8.9, 6.8 Hz), 7.43 (d, 1H, J=1.2 Hz), 7.35 (dd, 1H, J=8.2, 1.7 Hz), 7.25 (d, 1H, J=8.2 Hz), 6.53 (dd, 1H, J=11.8, 2.4 Hz), 6.47-6.42 (m, 1H), 2.25 (s, 3H), 0.26 (s, 9H); '*C-NMR (100 MHz; CDCls) § 172.86, 167.61 (d, Jc

F=253.3), 151.24 (d, Jc¢=12.3 Hz), 138.28, 135.38 (d, Jc.Fr=11.4 Hz), 134.85, 132.82, 130.63, 123.81, 119.91, 106.63, 105.23 (d, Jc.r=22.8 Hz), 104.77, 99.98 (d, Jc.k=26.7 Hz), 94.05, 17.78, 0.00; MS (APCI+) 342 (M+1, 100); IR (KBr) 2151, 1661, 1249 cm: Anal. calcdffound for: CigHzoFNO2Si C, 66.83/67.02; H, 5.90/6.00; N, 4.10/4.09; F, 5.56/5.45.

EXAMPLE 6B 4-Fluoro-2-(2-methyl-4-ethynyl-phenylamino)-benzoic acid (7).

To a solution of 6 in CH30H (30 ml), aqueous 1N KOH (10 ml) was added. After stirring at room temperature for 16 hours, the CH;OH was removed and the aqueous layer was acidified with 6N HCI (Scheme 2B). The resulting white precipitation was extracted in to Et,0, the Et,0O layer was dried (MgSO4) and concentrated to give 7 as tan colored solid. Yield 0.4274 g (91%); mp 177-178 °C; "H-NMR (400 MHz; CDCl) § 9.35 (s, 1H), 8.08-8.04 (m, 1H), 7.44 (s, 1H), 7.38-7.25 (m, 2H), 6.57 (d, 1H, J=11.8 Hz), 6.48-6.44 (m, 1H), 3.08 (s, 1H), 2.27 (s, 3H); *C-NMR (100 MHz; CDCls) § 172.84, 167.61 (d, Jc#=253.3), 151.15 (d, Jcr=12.3 Hz), 138.63, 135.40 (d, Jc.r=12.3

Hz), 135.00, 132.87, 130.81, 123.76, 118.79, 106.75, 105.33 (d, Jc.r=22.8

Hz), 100.03 (d, Jc#=26.0 Hz), 83.37, 17.83, 0.00; '*F-NMR (376 MHz; CDCl) 8 -101.24 to -101.31 (m); MS (APCI+) 270 (M+1, 100); IR (KBr) 3315, 1672, 1594, 1253 cm™; Anal. calcdffound for: CygH12FNO, C, 71.37/71.08; H, 4.49/4.82; N, 5.20/5.09.

EXAMPLE 7B 1-(4-nitro-phenyl)-1H-pyrrole (9a)

To a gently refluxing mixture of 4-nitroaniline (6.906 g, 0.05 mole), and sodium acetate (23 g, 0.28 mole) in acetic acid (100 ml) was added 2,5- dimethoxytetrahydrofuran (7.26 g, 7.12 ml, 0.055 mole) dropwise (Scheme 3B). After refluxing for 3 hours, the reaction mixture was poured on to crushed ice (~250 ml), basified with 10 % sodium hydroxide (250 ml) and extracted with CH,Cl,. The CHCl, layer was dried (K>CO3) to afford the product as a dark brown oil. Yield 9.40 g (100 %).

EXAMPLE 8B 1-(4-nitro-phenyl)-1H-pyrazole (Sb)

A mixture of pyrrazole (6.808 g, 0.1 mole) tetrabutylammonium bromide (3.22 g, 0.01 mole) and KOH (11.22 g, 0.2 mole) were ground together and sonicated for 16 hours. To this 1-fluoro-4-nitrcbenzene (15.521 g, 11.67 mi,

0.11 mole) was added and the mixture sonicated for 24 hours. The reaction mixture was extracted with CH2Cl,. The CH,Cl; layer was dried (MgSO,) and concentrated to give dark brown solid. This was purified by silica column chromatography. Elution with CH,Cl, (Rf = 0.44) gave the product as a light brown solid. Yield 8.80 g (47 %); mp 171-172 °C; Anal. calcd/found for: "

CgH;N3O; C, 57.14/56.52; H, 3.73/3.62; N, 22.21/21.95.

EXAMPLE 9B 3.5-dimethyl-1-(4-nitro-phenyl)-1H-pyrazole (9c)

To a solution of 4-nitro-phenyl-hydrazine (15.3 g, 0.1 mole) and 2,4- pentanedione (10.01 g, 10.27 ml, 0.1 mole) in EtOH (200 ml) were added 5 drops of concentrated HCI. The mixture was refluxed for 15 minutes; and the solvent removed to give a gummy product. This was purified by silica column chromatography. Elution with CH,Cl, gave the desired product (R; = 0.10) as abrown solid. Yield 7.22 g (33 %).

EXAMPLE 10B 4-Pyrrol-1-yl-phenylamine (10a)

Catalytic reduction (H2/RaNi (6 g) /THF) of 1-(4-nitro-phenyl)- 1H- pyrrole (9.69 g, 0.05149 mole) at 51 psi gave crude product as an oil (Scheme 3B). The product was purified by silica column chromatography. Elution with

CH.Cl, (Rf = 0.13) gave the pure product as white solid. Yield 8.06 g (99 %); mp 80-81 °C. :

EXAMPLE 11B

In a manner similar to the preparation of 4-pyrrol-1-yl-phenylamine, the following were prepared: 4-1H-Pyrazol-1-yl-phenylamine (10b). Dark brown oil, yield 6.26 g (100 %).

Benzenamine, 4-(3.5-dimethyl-1H-pyrazzol-1-yl) (10c). Dark brown oil.

Yield 6.45 g (100 %).

EXAMPLE 12B 4-Fluoro-2-(4-pyrrol-1-yl-phenylamino)-benzoic acid (11a)

To a solution of 4-pyrrol-1-yl-phenylamine (3.16 g, 0.02 mole) in THF (30 ml) at 778°C, a solution of LDA (2M, 15 ml, 0.03 mole) was added and the mixture stirred for 30 minutes. Solid 2,4-difluorobenzoic acid was added and the reaction mixture stirred for 16 hours as it warmed up to room temperature.

The solvent was removed and ether (100 ml) added to the dark oily residue.

This was stirred vigorously and the insoluble gummy precipitate separated by filtration. The gamy residue was dissolved in HzO, acidified to pH 1 with 10% - HCI, and extracted with Et;O. The Et,0 layer was dried (MgSO,) and concentrated to give the target compound as a brown solid. Yield 2.74 g (93 %); mp 223-225 °C (DEC); **F-NMR (376 MHz; CDCl) & -101.44 (s); MS (APCI+) 297 (M+1, 100); IR (KBr) 1658, 1526, 1254 cm™'.

In a manner similar to the preparation of 4-Fluoro-2-(4-pyrrol-1-yl- phenylamino)-benzoic acid, the following were prepared: 4-Fluoro-2-(4-pyrazol-1-yl-phenylamino)-benzoic acid (11b). Light brown solid, mp 212-213 °C. 2-[4-(3,5-Dimethyi-pyrazol-1-yl)-phenylamino]- 4-Fluoro benzoic acid (11c). Tan powder, mp 198 —200 °C.

EXAMPLE 1C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-5-dimethylsulfamoyl-3.4- difluoro-benzoic acid methyl ester (APK IC5,=222 nM) ’ Step a: Preparation of 1-dimethylsulfamoyl-2,3.4-trifluorobenzene

To a gently stirring solution comprised of 2,3,4-trifluorobenzenesulfonyl N chloride (5.70 g, 0.0247 mol) in 1,2-dichloroethane (200 ml) was introduced by bubbling gaseous anhydrous dimethylamine. The mixture became cloudy after several minutes and was subsequently washed with water (200 ml), 6 N aqueous hydrochloric acid (200 ml), brine (200 ml), was dried over anhydrous magnesium sulfate, and was concentrated in vacuo to obtain a yellow oil. The crude product was purified by flash chromatography. Elution with dichloromethane afforded 3.40 g of a white solid; 58 % yield; ‘H-NMR (400

MHz; CDCl) § 7.63-7.56 (m, 1H), 7.12-7.04 (m, 1H), 2.812 (s, 3H), 2.807 (s, 3H); "*F-NMR (376 MHz; CDCl3) § —124.91 to —125.03 (m), —127.98 to —- 128.03 (m), —156.41 to —156.53.

Step b: Preparation of 5-dimethylsulfamoyl-2,3,4-trifluoro-benzoic acid

To a cold (-78 °C) stirring solution comprised of 1-dimethylsulfamoyl- 2,3 4-trifluorobenzene in anhydrous tetrahydrofuran (60 ml) under a nitrogen atmosphere was added a commercially available lithium diisopropylamide solution (Aldrich, 2.0 M in tetrahydrofuran/heptane/ethylbenzene, 7.5 mi, 0.0150 mol). After stirring for about ten minutes, the purple solution was transferred via canula to a cold, stirring, saturated carbon dioxide in diethyl ether solution (200 ml). The reaction mixture took on a dull burgundy color.

The cold bath was removed and the reaction mixture warmed to ambient temperature over one hour. The mixture was then carefully quenched with 10 % aqueous hydrochloric acid (200 mi). The layers were separated. The organic phase was extracted twice (200, 100 ml portions) with 10 % (wt.) aqueous sodium hydroxide. The combined aqueous alkaline extracts were treated with concentrated aqueous hydrochloric acid (100 mi) to pHO. A white precipitate formed. The suspension was allowed to cool, then was extracted with diethyl ether (600 ml). The organic extract was dried over anhydrous magnesium sulfate and was concentrated in vacuo to afford 2.70 g of an off-white solid; 67.5 % yield; mp 225-228 °C; "H-NMR (400 MHz;

DMSO) 6 14.08 (broad s, 1H), 8.02-7.97 (m, 1H), 2.75 (s, 3H), 2.74 (s, 3H)

SF-NMR (376 MHz; DMSO) § -122.50 to ~122.63 (m), —122.95 to —123.08 (m), —154.49 to -154.61 (m); MS (APCI+) 284 (M+1, 22), 238 (100); (APCI-) 282 (M-1, 85), 259 (94), 238 (46), 216 (91), 195 (100); IR (KBr) 1702 cm™;

Anal. calcd/found for: CoHgF3NO4LS C, 38.17/38.40; H, 2.85/2.90; N, 4.95/4 80;

F, 20.12/19.75; S, 11.32/11.12.

Step c: Preparation of 5-dimethylsulfamoyl-2,3 4-trifluoro-benzoic acid methyl 10 . sete The solid 5-dimethylsulfamoyl-2,3,4-triflucro-benzoic acid (1.47 g, 0.00519 mol) and p-toluenesulfonic acid catalyst (17.1 mg) were dissolved in methanol (125 ml). The stirring mixture was brought to reflux under a nitrogen atmosphere for 51 hours. The reaction mixture was concentrated in vacuo to give a solid. The product was partitioned between diethyl ether (200 m!) and saturated aqueous potassium carbonate (75 ml). The layers were separated and the organic phase was washed with water (75 ml), brine (75 ml), was dried over anhydrous potassium carbonate, and was concentrated in vacuo to afford 0.15 g of an off-white solid; 10 % yield; "H-NMR (400 MHz; CDCl3) & 8.23-8.19 (m, 1H), 3.92 (s, 3H), 2.83 (s, 6H); "*F-NMR (376 MHz; CDCl3) & — 120.79 to —121.02 (m), —153.69 to —153.80.

Step d: Preparation of 2-(2-chloro-4-iodo-phenylamino)-5-dimethylsulfamoyl- 3.4-diflucro-benzoic acid methyl ester

To a stirring cold (-78 °C) solution comprised of 2-chloro-4-iodoaniline (0.143 g, 5.64x10* mol) in anhydrous tetrahydrofuran (5 ml) under a nitrogen atmosphere was added a commercially available lithium diisopropylamide solution (Aldrich, 2.0 M in tetrahydrofuran/heptane/ethylbenzene, 0.300 ml, 6.0x10™* mol). After stirring for 5 minutes, a solution comprised of 5- dimethylsulfamoyl-2,3,4-trifluoro-benzoic acid methyl ester (0.15 g, 5.0x10* mol) in tetrahydrofuran (10 ml) was added via syringe. The cold bath was removed and the reaction mixture was stirred for 2 hours. The reaction mixture was then partitioned between diethyl ether (125 ml) and saturated aqueous sodium bicarbonate (125 ml). The aqueous bicarbonate phase was extracted with an additional portion (125 ml) of diethyl ether. The combined organic phases were dried over anhydrous magnesium sulfate and concentrated in vacuo to give a yellow oil. The oil was crystallized from heptane-ethyl acetate to afford 0.060 g of an off-white powder; 23 % yield; mp 154-156 °C; "H-NMR (400 MHz; CDCl3) 5 9.74 (s, 1H), 8.30 (d, 1H, J=7.1 Hz), 7.72 (s, 1H), 7.49 (d, 1H, J=8.3 Hz), 6.73-6.69 (m, 1H), 3.92 (s, 3H), 2.84 (s, 3H), 2.83 (s, 3H); "F-NMR (376 MHz; CDCl;) § =123.90 (d), =139.55 (d).

EXAMPLE 2C 1 Preparation of 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3.4- difluoro-5-sulfamoyl-benzamide (PD 219622)

Step a: Preparation of 1-bis-(4-methoxybenzyl)sulfamoyl-2,3 4- trifluorobenzene

To a stirring solution comprised of bis-4-methoxybenzylamine (2.5 g, 9.7x10° mol) and diisopropylethylamine (1.7 mi, 9.7x10° mol) in dichloromethane (50 ml) at 0 °C under nitrogen atmosphere was added liquid 2,34- ’ trifluorobenzenesulfonyl chloride (2.26 g, 9.5x10 mol) directly. The mixture was stirred cold for ten minutes. The ice-water bath was removed and the mixture was stirred for an additional 15 minutes and was then diluted with dichloromethane to 350 ml volume and was washed with saturated aqueous ammonium chloride (200 ml). The organic phase was dried (MgSQOs) and concentrated in vacuo to afford 4.99 g of a sticky white solid. The crude product was recrystallized from hexanes-acetone to afford 3.00 g of white needles; 70 % yield; mp 87-90 °C; "H-NMR (400 MHz; CDCl;) § 7.64-7.58 (m, 1H), 7.04-6.99 (m, 1H), [6.97 (d, 4H, J=8.5 Hz), 6.75 (d, 4H, J=8.8 Hz) AB q], 4.33 (s, 4H), 3.76 (s, 6H); ®F-NMR (376 MHz; CDCl) § —125.44 to —125.56 (m), — 128.61 to —128.72 (m), —156.91 to —-157.03 (m); MS (APCI+) 121 (M-330, 100); (APCI-) 330 (M-121, 18), 195 (M-256, 100); IR (KBr) 1612, 1517, 1506, 1465, 1258, 1240, 1156, 1037, 1030 cm”; Anal. calcd/found for:

Ca2H20F3NO4S C, 58.53/57.98; H, 4.47/4.61; N, 3.10/2.85.

Step b: Preparation of 5-bis-(4-methoxybenzyl)sulfamoyl-2 3 4-trifluorobenzoic acid

To a stirring solution comprised of 1-bis-(4-methoxybenzyl)sulfamoyl-

S 2,3,4-trifluorobenzene (2.95 g, 6.5x10 mol) in tetrahydrofuran (60 ml) at -78 °C was added a solution comprised of 2.0 M lithium diisopropylamide in tetrahydrofuran/heptane/ethylbenzene (Aldrich, 3.35 ml, 6.7x10 mol). After several minutes of stirring, the dark solution was transferred via canula over five minutes to a stirring solution comprised of carbon dioxide (excess) in diethyl ether at -78 °C. A white precipitate immediately formed. The cold bath was removed and the reaction mixture was stirred at ambient temperature for 18 hours. The reaction mixture was quenched with 200 mi of dilute aqueous hydrochloric acid. The layers were separated and the organic phase was dried (MgSO,) and concentrated in vacuo to give 2.82 g of an off- white solid. Recrystallization from dichloromethane (150 ml) afforded 2.10g of the white powder product; 65 % yield; mp 158-161 °C; "H-NMR (400 MHz;

DMSO) § 7.80-7.76 (m, 1H), 7.05-6.74 (AB q, 8H, J=8.6 Hz), 4.33 (s, 4H), 3.66 (s, 6H); '°F-NMR (376 MHz; DMSO) 5 —123.28 to —123.36 (m), —124.12 to —124.21 (m), -155.41 to —155.53 (m); MS (APCI-) 494 (M-1, 47), 216 (89), 195 (100); IR (KBr) 3420, 2954, 2838, 1695, 1613, 1512, 1347, 1238, 1152, 1079 cm; Anal. calcd/found for: Ca3H20F3NOsS C, 55.76/55.85; H, 4.07/4.02;

N, 2.83/2.71; F, 11.50/11.41; S, 6.47/6.25.

Step c: Preparation of 5-bis-(4-methoxybenzyl)sulfamoyl-2-(2-chloro-4- iodophenylamino)-3 4-difluorobenzoic acid (PD 215729)

To a stirring solution comprised of 2-chloro-4-iodoaniline (0.53 g, 2.0x10"* mol) in tetrahydrofuran (10 ml) at —78 °C under a nitrogen atmosphere was added a solution comprised of 1.0 M lithium bis(trimethylsilyl)amide in tetrahydrofuran (Aldrich, 4.1 ml, 4.1x10™ mol).

Within several minutes the solution became a thick light-green suspension.

To this mixture was added a solution comprised of lithium 5-bis-(4- methoxybenzyl)sulfamoyl!-2,3,4-trifluorobenzoate in tetrahydrofuran, which was prepared by adding 2.0 ml of the Aldrich lithium bis(trimethylsilyl)amide solution (0.0020 mmol) to a solution comprised of 5-bis-(4-

methoxybenzyl)sulfamoyl-2,3,4-trifluorobenzoic acid (1.00 g, 2.0x1 0° mol) in tetrahydrofuran (10 ml) at —78 °C. The reaction mixture was stirred for 15 minutes and was then concentrated in vacuo to a crude semisolid. The semisolid was taken up into diethyl ether (250 ml) and was washed with 1 % aqueous hydrochloric acid (150 mi). The ether phase was then washed with neutral water (200ml, pH 4 after wash), a second portion of water (200 mi, pH 6 after wash), and brine (200 ml). The organic phase was then dried (MgSO) and was concentrated in vacuo to give 1.88 g of a sticky residue which was crystallized from toluene-heptane to afford 1.12 g of an off-white powder; 76 % yield; mp 162-166 °C; 'H-NMR (400 MHz; DMSO) 5 9.86 (s, 1H), 7.92 (d, 1H, J=6.8 Hz), 7.86 (d, 1H, J=1.7 Hz), 7.60 (dd, 1H, J=8.5, 1.7 Hz), 7.06- 7.04/6.78-6.75 (AB q, 8H, J=8.5 Hz), 6.93-6.89 (m, 1H), 4.31 (s, 4H), 3.66 (s, 6H); '9F-NMR (376 MHz; DMSO) § —127.22 (d), —141.36 (d); MS (APCI+) 729 (M+1, 1), 256 (50), 121 (100); (APCI-) 727 (M-1, 100); IR (KBr) 1698, 1673, 1513, 1251 cm™; Anal. calcd/found for: C9H24CIF2IN2O6S C, 47.78/47.93; H, 3.32/3.33; N, 3.84/3.80; Cl, 4.86/4.84; F, 5.21/5.46; 1, 17.41/17.16; S, 4.40/4.29.

Step d: Preparation of 5-bis-(4-methoxybenzyl)sulfamoyl-2-(2-chloro-4- iodophenylamino)-N-cyclopropyimethoxy-3,4-diflucrobenzamide (PD 218774)

To a stirring solution comprised of 5-bis-(4-methoxybenzyl)sulfamoyl-2- (2-chloro-4-iodophenylamino)-3,4-difluorobenzoic acid (0.935 g, 1.28x1 0? mol), cyclopropylmethoxylamine hydrochloride (0.175 g, 1.42x10° mol), and diisopropylethylamine (0.75 mi, 4.26x10™ mol) in a 1:1 v/v tetrahydrofuran- dichloromethane mixture (50 ml) was added solid PyBOP ([benzotriazolyloxy]tripyrrolidino phosphonium hexafluorophosphate,

Advanced ChemTech, 0.76 g, 1.46x10 mol). The reaction mixture was stirred for one hour, was then evaporated to a crude residue which was purified by flash silica column chromatography. Elution with a gradient (25 % dichloromethane to 75 % dichloromethane in hexanes) afforded 0.63 g of the off-white powder product; 62 % yield; mp 70->300 °C; 'H-NMR (400 MHz;

DMSO) & 11.92 (s, 1H), 9.35 (s, 1H), 7.60 (s, 1H), 7.50-7.45 (m, 1H), 7.34 (d, 1H, J=8.5 Hz), 6.82-6.54 (AB q, 8H, J=8.3 Hz), 6.59-6.54 (m, 1H), 4.09 (s,

4H), 3.46 (s, 6H), 0.90-0.80 (m, 1H), 0.30-0.25 (m, 2H), 0.03-0.00 (m, 2H);

YF.NMR (376 MHz; DMSO) & -129.05 (s), —140.23 (d, J=18.3 Hz); MS (APCI+) 798 (M+1, 70), (APCI-) 796 (M-1, 15), 726 (50), 131 (100); IR (KBr) 1642, 1611, 1584, 1513, 1478 cm"; Anal. calcd/found for: C33H3iCIF2IN3O6S

C,649.67/49.88; H, 3.92/3.95; N, 5.27/5.19.

Step e: Preparation of 2-(2-chloro-4-iodophenylamino)-N-cyclopropylmethoxy- 3.4-difluoro-5-sulfamoyl-benzamide (PD 219622)

A reaction solution comprised of 5-bis-(4-methoxybenzyl)sulfamoyl-2- (2-chloro-4-iodophenylamino)-N-cyclopropylimethoxy-3,4-difluorobenzamide (0.1010 g, 1.266x10™ mol) in trifluoroacetic acid (4 mi) was stirred at ambient temperature for 24 hours. The mixture was vacuum filtered and the precipitate rinsed with hexanes to afford 28.6 mg of a pale lavender powder; 42 % yield; mp 219-227 °C DEC; "H-NMR (400 MHz; DMSO) 5 11.89 (s, 1H), 9.08 (s, 1H), 7.60 (s, 3H), 7.55 (d, 1H, J=6.9 Hz), 7.32 (d, 1H, J=8.6 Hz), 6.63- 6.59 (m, 1H), 3.40 (d, 2H, J=6.6 Hz), 0.90-0.80 (m, 1H), 0.30-0.26 (m, 2H), 0.05-0.00 (m, 2H); "*F-NMR (376 MHz; DMSO) & -130.61 (s), —140.38 (d, J=21.4 Hz), MS (APCI+) 558 (M+1, 70), 282 (100); (APCI-) 556 (M-1, 73), 486 (100); IR (KBr) 3390, 3283, 1652, 1513, 1477, 1163 cm’: Anal. calcd/found for: C47H;5CIF2IN3O4S 0.1 C,HF30, C, - 36.30/36.31; H, 2.67/2.55; N, 7.38/7.00.

EXAMPLE 3C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-3.4-difluoro-N-hydroxy-5- sulfamoyl-benzamide (PD 224213)

To a stirring solution comprised of 5-bis-(4-methoxybenzyl)sulfamoyl-2- (2-chloro-4-iodophenylamino)-3,4-difluorobenzoic acid (0.67 g, 9.2x1 0 mol),

O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (0.113 g, 9.65x10™ mol), and diisopropylethylamine (0.50 ml, 2.9x10™ mol) in a 1:1 v/v tetrahydrofuran- dichloromethane mixture (20 ml) was added solid PyBOP ([benzotriazolyloxyltripyrrolidino phosphonium hexafluorophosphate,

Advanced ChemTech, 0.52 g, 1.0x10 mol). The reaction mixture was stirred for 30 minutes, was concentrated in vacuo to a yellow oil, and was crystallized from methanol to afford 0.35 g of the off-white amorphous intermediate; 46 %

yield; the intermediate was dissolved in trifluoroacetic acid (10 ml) and was stirred at ambient temperature for 16 hours. The mixture was vacuum filtered to collect the precipitate, which was recrystallized from methanol-chloroform to afford 0.055 g of the tan powder product; 26 % yield from intermediate; mp 230-236 °C DEC; 'H-NMR (400 MHz; DMSO) 3 11.73 (s, 1H), 9.46 (s, 1H), 9.38 (s, 1H), 7.80-7.75 (m, 2H), 7.79 (s, 2H), 7.50 (d, 1H, J=8.5 Hz), 6.82-6.78 (m, 1H); *°F-NMR (376 MHz; DMSO) 5 —130.83 (s), —139.24 (s); MS (APCI+) 504 (M+1, 53), 488 (90), 471 (100); (APCI-) 502 (M-1, 12), 486 (100); IR (KBr) 3295, 1652, 1636, 1519, 1477, 1315, 1157 cm™; Anal. calcd/found for: Cy3HGCIF2IN3O4S “0.41 CHCl; C, 29.15/29.05; H, 1.72/1.66; N, 7.60/7.21.

EXAMPLE 4C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5-sulfamoyi- benzoic acid (PD 215730)

Solid 5-bis-(4-methoxybenzyl)sulfamoyl-2-(2-chloro-4- } iodophenylamino)-3,4-difluorobenzoic acid (0.0995 g, 1.36x10™ mol) was dissolved in trifluoroacetic acid (5 ml) under a nitrogen atmosphere. The reaction mixture was stirred at ambient temperature for 65 hours. The mixture was vacuum filtered to isolate 55.2 mg of a fine white precipitate. The crude product was recystallized from chloroform to afford 31.8 mg of the fluffy white solid product; 48 % yield; mp 295-296 °C DEC; "H-NMR (400 MHz; DMSO) § 9.77 (s, 1H), 8.16 (d, 1H, J=7.3 Hz), 7.82 (s, 3H), 7.56 (d, 1H, J=8.5 Hz), 6.97- 6.92 (m, 1H); 'SF-NMR (376 MHz; DMSO) § —128.47 (s), —141.13 (d, 19.8

Hz); MS (APCI+) 489 (M+1, 5), 102 (100); (APCI-) 491 (32), 490 (18), 489 (100), 488 (18), 487 (M-1, 75); IR (KBr) 3372, 3244, 1688 cm™; Anal. calcd/ffound for: C13HsCIF2IN204S C, 31.96/32.19; H, 1.65/1.81; N, 5.73/5.37.

EXAMPLE 5C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3.4- difluoro-5-dimethyisulfamoyl-benzamide (PD 250253)

Step a: Preparation of 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5- dimethylsulfamoyl-benzoic acid (PD 224339)

To a stirring solution comprised of 5-dimethylsulfamoyl-2,3,4- trifluorobenzoic acid (1.00 g, 3.53x107 mol) in tetrahydrofuran (15 ml) at —78 °C under a nitrogen atmosphere was added a 1.0 M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (Aldrich, 3.6 ml, 3.6x10° mol). A lithium 2-chloro-4-iodoanilide suspension formed by adding a 1.0 M solution of lithium bis(trimethylsilyl)amide solution (7.2 ml, 7.2x10® mol) to a solution comprised of 2-chloro-4-iodoaniline (0.94 g, 3.63x10"2 mol) in tetrahydrofuran (15 ml) at -78 °C was added via canula to the lithium 5-dimethylsulfamoyl- 2,3 4-trifluorobenzoate suspension. The cold bath was removed and the reaction mixture was stirred for one hour. The mixture was concentrated in vacuo to a crude solid. The crude product was suspended in diethyl ether (200 ml), to which suspension hydrogen chloride gas was introduced to produce a white precipitate. The precipitate was removed by vacuum filtration. The filtrate was concentrated in vacuo to give a dull-colored solid, which was triturated with hexanes-dichloromethane to afford 1.31 g of the white powder product; 72 % yield; mp 218-222 °C; "H-NMR (400 MHz;

DMSO) 8 9.89 (s, 1H), 8.06 (d, 1H, J=6.1 Hz), 7.85 (d, 1H, J=1.9 Hz), 7.58 (dd, 1H, J=8.5, 1.9 Hz), 7.03 (dd, 1H, J=8.3, 6.6 Hz), 2.71 (s, 6H); *F-NMR (376 MHz; DMSO) 6 -125.58 (d, J=18.3 Hz), -140.14 (d, J=16.8 Hz); MS (APCI+) 519 (40), 518 (15), 5617 (M+1, 100); (APCI-) 517 (6), 516 (2), 515 (M- 1, 5), 480 (45), 127 (100); IR (KBr) 3346, 1665, 1487, 1283 cm; Anal. calcd/found for: C15sH12CIF2IN204S C, 34.87/34.98; H, 2.34/2.32; N, 5.42/5.32.

Step b: Preparation of 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-5-dimethyisulfamoyl-benzamide

To a suspension comprised of 2-(2-chloro-4-iodo-phenylamino)-3,4- difluoro-5-dimethylsulfamoyl-benzoic acid (0.5 g, 9.68x10™ mol) and cyclopropyimethoxylamine hydrochloride (0.13 g, 1.05x10 mol) in a 1:1 viv mixture of dichloromethane-tetrahydrofuran (10 ml) was added diisopropylethylamine (0.65 ml, 3.73x1 03 mol) followed by the addition of solid

PyBOP (0.55 g, 1.06x10 mol). The reaction mixture was stirred at ambient temperature for three days. The mixture was concentrated in vacuo to a red oil. The crude product was treated with 10 % aqueous hydrochloric acid (150 mi) and was extracted with diethyl ether (150 ml). The organic phase was dried (MgSO,) and concentrated in vacuo to a crude solid. The solid was triturated with dichloromethane-hexanes and recovered by vacuum filtration to afford 0.3558 g of the white powder product; 63 % yield; mp 222-225 °C DEC;

H-NMR (400 MHz; DMSO) § 11.97 (s, 1H), 9.32 (s, 1H), 7.60 (d, 1H, J=1.9

Hz), 7.49 (d, 1H, J=5.8 Hz), 7.33 (dd, 1H, J=8.4, 1.9 Hz), 6.70 (dd, 1H, 8.4, 6.3 Hz), 3.43 (d, 2H, J=7.2 Hz), 2.53 (s, 6H), 0.87-0.83 (m, 1H), 0.30-0.25 (m, 2H), 0.03-0.00 (m, 2H); 'F-NMR (376 MHz; DMSO) § —127.67 (d, J=19.8 Hz), -139.32 (d, J=19.8 Hz); MS (APCI+) 586 (M+1, 100); (APCI-) 584 (M-1, 40), 514 (100); IR (KBr) 3263, 1644, 1585, 1507, 1480 cm’: Anal. calcd/found for:

C19H19CIF2IN30,4S C, 38.96/39.08; H, 3.27/3.18; N, 7.17/7.17.

EXAMPLE 6C

Preparation of N-cyclopropylmethoxy-3,4-difluoro-5-dimethylsulfamoyl-2-(4- iodo-2-methyl-phenylamino)-benzamide (PD 252745)

Step a: Preparation of 3 4-difluoro-5-dimethylsulfamoyl-2-(4-iodo-2-methyl- phenylamino)-benzoic acid (PD 224340)

Same procedure and same scale as Example 4C, Step a, except 4- iodo-2-methylaniline was used instead of 2-chloro-4-iodoaniline; afforded 0.9592 g of the off-white powder product; 55 % yield; mp 235-238 °C; "H-NMR (400 MHz, DMSO) & 9.69 (s, 1H), 8.04 (d, 1H, J=6.1 Hz), 7.60 (d, 1H, J=1.5 i Hz), 7.45 (dd, 1H, J=8.3, 1.7 Hz), 6.88 (dd, 1H, J=8.3, 5.4 Hz), 2.70 (s, 6H), 2.21 (s, 3H); "*F-NMR (376 MHz; DMSO) § —126.25 (d, J=16.8 Hz), -142.74 (d, J=19.8 Hz); MS (APCI+) 497 (M+1, 69), 357 (70), 316 (100); (APCI-) 495 (M-1, 3), 127 (100); IR (KBr) 3240, 1686, 1512, 1473, 1341, 1151 cm’; Anal. calcd/found for: C4eH15F2IN2O4S C, 38.72/38.70; H, 3.05/3.01; N, 5.64/5.49.

Step b: Preparation of N-cyclopropylmethoxy-3.4-difluoro-5- dimethylsulfamoyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

Same procedure and same scale as Example 4C, Step b, except the product was purified by recrystallization from absolute ethanol to afford 0.1718 g of the pale yellow microcrystalline product; 28 % yield; mp 171-172 °C; H-

NMR (400 MHz; DMSO) 8 11.79 (s, 1H), 8.91 (s, 1H), 7.40 (d, 1H, J=4.3 Hz),

7.36 (s, 1H), 7.21 (d, 1H, J=8.2 Hz), 6.54 (dd, 1H, 8.2, 4.3 Hz), 3.30 (d, 2H,

J=6.5 Hz), 2.52 (s, 6H), 2.00 (s, 3H), 0.85-0.75 (m, 1H), 0.29 (d, 2H, J=7.7

Hz), 0.01 (d, 2H, J=4.1 Hz); '°F-NMR (376 MHz; DMSO) 5 —128.94 (s), — 143.32 (d, J=19.8 Hz); MS (APCI+) 566 (M+1, 100); (APCI-) 564 (M-1, 85),

S 494 (100); IR (KBr) 1649, 1609, 1588, 1512, 1475 cm; Anal. calcd/found for:

CaoH22F2IN304S C, 42.49/42.42; H, 3.92/3.78; N, 7.43/7.40.

EXAMPLE 7C

Preparation of 2-(2-chloro-4-iodo-phenylamino)-N-hydroxy-4- dimethylsuifamoyl-benzamide

Step a: Preparation of 4-methyl-benzene-N.N-dimethylsulfonamide

To a stirring solution comprised of para-toluenesulfonyl chloride in dichloromethane at 0 °C is introduced excess gaseous dimethylamine. The precipitate is removed by filtration and the filtrate is concentrated in vacuo to obtain the product.

Step b: Preparation of 4-methyl-3-nitro-benzene-N,N-dimethyisulfonamide

To a gently stirring solution comprised of 1 molar equivalent of fuming nitric acid in excess concentrated sulfuric acid is added 1 molar equivalent of 4-methyl-benzene-N,N-dimethylsulfonamide in increments. The mixture is stirred for one hour and then poured over chilled water. The mixture is extracted with a suitable solvent like diethyl ether or dichloromethane. The organic phase is dried over a suitable drying agent like magnesium sulfate and concentrated in vacuo to afford a crude product which may be purified by normal methods such as chromatography or crystallization from a solvent like chloroform or heptane.

Step c: Preparation of 3-amino-4-methyl-benzene-N,N-dimethylsulfonamide

The compound 4-methyl-3-nitro-benzene-N,N-dimethylsulfonamide is dissolved in ethanol. A catalyst like Raney nickel is added and the mixture hydrogenated in a shaker. The catalyst is removed by filtration. The solvent is removed in vacuo to give a product which may be purified if necessary by chromatography or crystallization from an appropriate solvent like chloroform or heptane-ethyl acetate.

Step d: Preparation of 3-fluoro-4-methyl-benzene-N,N-dimethylsulfonamide

The compound 3-amino-4-methyl-benzene-N,N-dimethylsuifonamide is diazotized with an alkyl nitrite like tert-butyl nitrite under anhydrous conditions in a non-reactive solvent like tetrahydrofuran or dichloromethane. The intermediate diazonium species is then treated with pyridinium fluoride to give the product, which may be purified by chromatography or crystallization.

Step e: Preparation of 4-dimethylsulfamoyl-2-fluoro-benzoic acid

A mixture comprised of 3-fluoro-4-methyl-benzene-N,N- dimethylsulfonamide and potassium permanganate (2.2 molar equivalents) in water is brought to reflux for four hours. The reaction mixture is filtered through celite. The filtrate is treated with activated carbon and refiltered through fresh celite. The second filtrate is acidified with concentrated hydrochloric acid to pH 0. The mixture is allowed to cool and is extracted with : diethyl ether. The organic phase is dried over a drying agent like magnesium sulfate and is concentrated in vacuo. The product may be purified by recrystallization from an appropriate solvent like ethanol or chloroform. .

Step f: Preparation of 2-(2-chloro-4-iodo-phenylamino)-4-dimethyisulfamoyl- benzoic acid

To a stirring cold (-78 °C) solution comprised of 2-chloro-4-iodoaniline (1 molar equivalent) in anhydrous tetrahydrofuran under a nitrogen atmosphere is added a commercially available lithium diisopropylamide solution (Aldrich, 2.0 M in tetrahydrofuran/heptane/ethylbenzene, 1 molar equivalent). After stirring for 5 minutes, a solution comprised of 4- dimethylsulfamoyi-2-fluoro-benzoic acid (1 molar equivalent) in tetrahydrofuran is added. The cold bath is removed and the reaction mixture is stirred for 2 hours. The reaction mixture is then partitioned between diethyl ether and dilute aqueous hydrochloric acid. The organic phase is washed with brine, dried over magnesium sulfate, and concentrated in vacuo to afford a product which may be purified by chromatography of recrystallization from an appropriate solvent like chloroform or heptane-ethanol.

Step g: Preparation of 2-(2-chloro-4-iodo-phenylamino)-4-dimethylsulfamoyl- benzoic acid O-(tetrahydro-2H-pyran-2-yl)-oxyamide

A solution comprised of 2-(2-chloro-4-iodo-phenylamino)-4- dimethylsulfamoyl-benzoic acid, O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (1.25 molar equivalents), benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (1.25 molar equivalents), and diisopropylethylamine (3 molar equivalents) in 1:1 v/v tetrahydrofuran-dichloromethane is stirred for 30 minutes. The reaction mixture is concentrated in vacuo and the residue is purified by flash chromatography; elution with dichloromethane affords the desired product. The product may be recrystallized with an appropriate solvent like methanol if further purification is necessary.

Step h: Preparation of 2-(2-chloro-4-iodo-phenylamino)-4-dimethyisulfamoy!-

N-hydroxy-benzamide

The compound 2-(2-chloro-4-iodo-phenylamino)-4-dimethylsulfamoyl- benzoic acid O-(tetrahydro-2H-pyran-2-yl)-oxyamide is dissolved in an appropriate hydrogen chloride-saturated solvent like methanol or ethanol.

Once homogeneous, the solution is concentrated in vacuo to give the desired product. The product may be triturated with an appropriate solvent like chloroform or dichloromethane if further purification is necessary.

F. Other Embodiments

From the above disclosure and examples, and from the claims below, the essential features of the invention are readily apparent. The scope of the invention also encompasses various modifications and adaptations within the knowledge of a person of ordinary skill. Examples include a disclosed compound modified by addition or removal of a protecting group, or an ester, pharmaceutical salt, hydrate, acid, or amide of a disclosed compound.

Publications cited herein are hereby incorporated by reference in their entirety.

What is claimed is:

Claims (1)

1. PCT/US00/18348 N | CLAIMS

   1. Use of a MEK inhibitor selected from: a compound of formula (1): Rig . } 0 NO Ry ~s® LR : CL Rs Rs l Oo Riis H, C 15 alkyl, C 35 alkenyl, C 35 alkynyl, C 55 cycloalkyl, phenyl, (phenyl)C 14 alkyl, (phenyl)C 34 alkenyl, (phenyl)C 1. alkynyl, (C 3.5 cycloalkyl)-C 14 alkyl, (C 33 cycloalkyl)C 34 alkenyl, (C 3.5 cycloalky)C 34 “alkynyl, C a6 heterocyclic radical, (C 34 heterocyclic radical)C ;4 alkyl, (C 18 i heterocyclic radical)C 34 alkenyl, (C 33 heterocyclic radical)C 14 alkynyl, (CH2)24 ORc or (CH2)2.4 NRcRp ; : Rz is H, C 14 alkyl, phenyl, C 35 cycloalkyl, C 35 heterocyclic radical, or (C 16 cycloalkyl) methyl; oo each of Rj; and Ry is independently selected from H, F , NOs, Br and CI; Rs is selected from H and F; Re is H, F, Cl or CHx; each of Rc and Rp is independently selected from H, C 14 alkyl, C 34 alkenyl, C 34 alkynyl, C 35 cycloalkyl, and phenyl; or NRcRp may be a piperidino, morpholino, or N-(C 4 alkyl)piperazino ring; 117 AMENDED SHEET

   PCT/US00/18348 wherein each hydrocarbon radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, hydroxyl, amino, (amino)sulfonyl, and NO,; and wherein each heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C , alkyl, C 35 : cycloalkyl, C ,, alkenyl, C ,, alkynyl, phenyl, hydroxyl, amino, (amino)sulfonyl, and NO,, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 2 substituents : independently selected from halo, C ,, alkyl, hydroxyl, amino, and NO,; or a pharmaceutically acceptable saltor C 1. ester thereof, in the manufacture of a medicament for treating chronic pain.

   2. Use of claim 1, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   3. Use of claim 2, wherein said chronic pain is a type of neuropathic pain.

   4. Use of claim 3, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other nerve injury between the peripheral nervous system and the central nervous system, inclusively.

   5. Use of claim 2, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism 118 AMENDED SHEET

   ® | | PCT/US00/18348

   6. Use of claim 2, wherein said chronic pain is associated with idiopathic pain.

   7. Use of claim 1, wherein said chronic pain is associated withinflammation.

   8. | Use of claim 1 wherein said chronic pain is associated with arthritis.

   S. Use of claim 1, wherein said chronic pain is associated with post- operative pain. . 10. Use of claim 1, wherein R; is rom or chloro.

   11. Use of claim 1, wherein R, is fluoro.

   12. Use of claim 1, wherein Rs is H. .

   13. Use of claim 13, wherein each of R, and Rs is H. | :

   14. Use of claim 1, whersin each of R, and R; is fluoro.

   15. | Use of claim 14, wherein R; is bromo.

   16. Use of claim 14, wherein R; is fluoro.

   17. Use of claim 1, wherein R, is nitro,

   18. Use of claim 16, wherein Rs is H.

   18. Use of claim 1, wherein R; is chloro. 119 AMENDED SHEET

   PCT/US00/18348

   20. Use of claim 1, wherein Rg is methyl. :

   21. Use of claim 1, wherein R, is Hor C , alkyl, and R, is H.

   22. Use of claim 1, wherein R, is (C 54 cycloalkyl) methyl.

   23. Use of claim 1, wherein R, is H.

   24. Use of claim 1, wherein R, is (CH,) ,, OR. or (CH,),4 NRcR,.

   25. Use of claim 1, wherein said MEK inhibitor has a structure selected from: 4-fluoro-2-(4-iodo-2-methyl-rhenylamino)-benzenesuifonic acid; 4-fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenytamino)- benzenesulfonamide; N-cyclopropyimethoxy-4-fluoro-2-(4-iodo-2-methyl- : phenylamino)-benzenesulfonamide; 3,4-difluoro-2-(4-iodo-2-methyi- phenylamino)-benzenesulfonic acid; 3,4-difluoro-N-hydroxy-2-(4-iodo-2- methyl-phenylamino)-benzenesulfonamide; N-cyclopropyimethoxy-3.4- difluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide; 3,4,5-trifluoro- 2-(4-iodo-2-methyl-phenylamino)-benzenesulfonic acid; 3,4,5-trifluoro-N- : hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide; N- cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)- benzenesulfonamide; 5-bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)- benzenesulfonic acid; 5-bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyi- phenylamino)-benzenesulfonamide; 5-bromo-N-cyciopropyimethoxy-3,4- difiuoro-2-(4-iodo-2-methyl-phenytamino)-benzenesuffonamide; 2-(4-iodo-2- methyl-phenylamino)-4-nitro-benzenesulfonic acid; N-hydroxy-2-(4-iodo-2- methyl-phenyiamino)-4-nitro-benzenesuffonamide; and - 30 N-cyclopropyimethoxy-2-(4-iodo-2-methyl-phenylaminc)-4-nitro- benzenesulfonamide. oo 120 AMENDED SHEET

   : PCT/US00/18348 ®

   26. Use of claim 1, wherein said MEK inhibitor has a structure selected from: 2-(2-chloro-4-iodo-phenylamino)-4-fluoro-benzenesulfonic acid; 2-(2-chloro-4-iodo-phenylamino)-4-fluoro-N-hydroxy- _ benzenesulfonamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy- 4-fluoro-benzenesulfonamide; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro- benzenesulfonic acid; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N- hydroxy-benzenesulfonamide; 2-(2-chloro-4-iodo-phenylamino)-N- cyclopropyimethoxy-3,4-difluoro-benzenesuifonamide; 2-(2-chioro-4-iodo- phenylamino)-3,4,5-trifluoro-benzenesulfonic acid; 2-(2-chloro-4-iodo- phenylamino)-3,4,5-trifluoro-N-hydroxy-benzenesulfonamide; 2-(2-chloro-4- : iodo-phenylamino)-N-cyclopropylmethoxy-3,4, 5-trifluoro-benzenesulfonamide; : 5-bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-diflucro-benzenesulfonic acid; 5-bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy- benzenesulfonamide; 5-bromo-2-(2-chloro-4-iodo-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide; 2-(2-chloro-4-iodo- phenylamino)-4-nitro-benzenesulfonic acid; 2-(2-chloro-4-iodo-phenylamino)- N-hydroxy-4-nitro-benzenesulfonamide; and 2-(2-chloro-4-iodo-phenytamino)- N-cyclopropylmethoxy-4-nitro-benzenesulfonamide.

   27. Use of a MEK inhibitor selected from a compound having the formula (INA:w__0 q R,

   Fr. i . | 1 Rg (IhA 121 . AMENDED SHEET

   PS | PCT/US00/18348 Wis ORjy, NR2OR;,, NRaRs.

   NR2NR4Rg, of NR2(CHz)4 NRaRE: : Riis H, C 15 alkyl, C 55 alkenyl, C 35 alkynyl, C 35 cycloalkyl, phenyi, : (phenyl)C 14 alkyl, (phenyl)C 3. alkenyl, (phenyl)C 5. alkynyl, (C 1¢ E cycloalkyl)- | | oo C14 alkyl, (C 3.5 cycloalkyl)C 3-4 alkenyl, (C 34 cycioalkyl)C 4 alkynyl, C 24 : heterocyclic radical, (C sg heterocyclic radical)C 14 alkyl, (C 3.5 heterocyclic : radical)C 34 alkenyl, (C ss heterocyclic radical)C 14 alkynyl or (CH2)24NRARE; Rz is H, phenyl, C 14 alkyl, Cs4 alkenyl, C 35 alkynyl, C 33 cycloalkyl, or (C 38 cycloalkyl)-C 1 alkyl; oo oo oo Rais H, C 1 alkyl, C 35 alkenyl, C 35 alkynyl, C sq cycloalkyl, phenyl, (C55 ~~ ~ cycloalkyl)C 14 alkyl, (C 35 cycloalkyl)C 14 alkenyl, (Css cycloalkyl)C 34 alkynyl, C ss heterocyclic radical, (C 35 heterocyclic radical)C 14 alkyl, (aminosuifonyl)phenyl, [(aminosuifony)phenyl]C 14 alkyl, (aminosulfonyl)C 16 alkyl, (aminosulfonyl)C a5 cycloalkyl, or [(aminosulfonyl)C 15 cycloalkyl]C 14: _— alkyl; I Rg is H, C 15 alkyl, C 35 alkenyl, C 55 alkynyl, C 35 cycloalkyl, or C ea aryl; . : Rs is halo, NO3, SO:NR((CH2)2«NReRF, SONR, Rk or (CO)T; Tis C 1 alkyl, C 34 cycloalkyl, (NReRr)C 14 alkyl, ORe, NR|(CH2)24NReRF, or : NReRe | 3 Rs is Hor F; | | - Rs is H, methyl, halo, or NO2;

   Rs is H, methyl, halo, or NO; : oo | AMENDED SHEET

   PCT/US00/18348 Ar is phenyl, 2-pyridyl, 3-pyridyl, or 4-pyridy!; each of R; and Rg is independently selected from H, halo, C alkyl, SO2NR, (CH2)24NRgRH, (CO) CH;)24NReRy, (CO)NRj(CH2)24NRRy, (CO)O(CH2)24NRgR#, SO2NRGRy, and (CO)NRgRy; provided that where Ar is a pyridyl, each of R; and Rg is H; each of Re, Rp, Re, Rr, Rg, and Ry is independently selected from H, C 1 alkyl, : 10 Casa alkenyl, C 34 alkynyl, C 35 cycloalkyl, and phenyl; each of NRcRp, : NReRg, and NRgRH can also be independently morpholinyl, piperazinyl, - pyrrolidinyl, or piperadinyi; each of Rand R; is independently H, methyl, or ethyl; ) Rk is C 14 alkyl, C 34 alkenyl, C 34 alkynyl, C 25 cycloalkyl, or phenyl; Xis O, S, or NH; and wherein each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 14 alkyl, C 36 cycloalkyl, C ,4 alkenyl, C o4 alkynyl, phenyl, hydroxyl, amino, (amino)sulfonyl, and NO,, wherein each substituent alkyl, cycloalkyt, alkenyl, alkynyl or phenyl is in tum optionally substituted with between 1 and 2 substituents independently selected from halo, C 1.; alkyl, hydroxyl, amino, and NO; or a pharmaceutically acceptable salt or C ,; ester thereof, in the manufacture of a medicament for treating chronic pain. 123 AMENDED SHEET

   PCT/US00/18348

   28. Use of claim 27, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   29. Use of claim 28, wherein said chronic pain is a type of neuropathic pain.

   30. Use of claim 29, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other nerve injury between the peripheral nervous system and the central nervous system, inclusively.

   31. Use of claim 28, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism © 32. Use of claim 28, wherein said chronic pain is associated with idiopathic pain.

   33. Use of claim 27, wherein said chronic pain is associated with inflammation.

   34. Use of claim 27 wherein said chronic pain is associated with arthritis.

   35. Use of claim 27, wherein said chronic pain is associated with post- operative pain. 124 AMENDED SHEET

   PCT/US60/18348

   36. Use of claim 27, wherein the MEK inhibitor has the following formula : (DA: CW _0 N T LC PN Za | NUN Ry™ YR 1 X R, (DA wherein | . Wis ORy, NR20OR;, NRARag, NRoNRARE, or NR2(CH,).4 NRARSg; a Riis H, C 1.5 alkyl, C 2g alkenyl, C 14 alkynyl, C 35 cycloalkyl, phenvt, (phenyi)C 14 alkyl, (phenyl)C 34 alkenyl, (phenyl)C 14 alkynyl, (C 38 : cycloalkyl)- oo C 14 alkyl, (C 34 cycloalkyl)C 34 alkenyl, (C 1s cycloalkyl)C 34 alkynyl, C 24 heterocyclic radical, (C 3.5 heterocyclic radical)C 14 alkyl, (C 14 heterocyclic radical)C 34 alkenyl, (C 3 heterocyclic radical)C 34 alkynyl or (CH,)>4<NRARs: oo 15 Rz is H, phenyl, C 1 alkyl, C3. alkenyl, C 54 alkynyl, C 34 cycloalkyl, or (C 3s cycloalkyl)-C 14 alkyi; Rais H, C 1 alkyl, C 35 alkenyl, C 35 alkynyl, C 14 cycloalkyl, phenyl, (C 18 : cycloalkyl)C 44 alkyl, (C 34 cycloalkyl)C 14 alkenyl, (C 33 cycloalkyl)C 34 alkynyl, C 3.5 heterocyclic radical, (C 35 heterocyclic radical)C 4 alkyl, : (aminosulfonyf)phenyl, [(aminosulfonyl)phenyl]C 1 alky!, (aminosulfonyl)C 16 125 AMENDED SHEET

   ® - ] PCT/US00/18348 : alkyl, (aminosulfonyl)C 15 cycloalkyl, or [(amincsulfonyl)C 35 cycloalkyl]C 4 alkyl; | | oo oo - Rg is H, C 1s alkyl, C35 alkenyl, C 3-8 alkynyl, C 35 cycloalkyl, or Cg aryl; : Rs is halo, NO3, SO,NR; (CHa);4NReR-, SO,NR, Rx or (CO)T; - TisCsalkyl, Css cycloalkyl, (NReRg)C 14 alkyl, ORF, NRi(CHz)2<NReRe, or NReRE: } ST .

   RiisHor F; - Lo ’

   Rs is H, methyl, halo, or NO; - Co oo N ~~ 15 Reis H, methyl, halo, or NO; . | : oo each of R7 and Rj is independently selected from H, halo, Cid alkyl, oo SONRy (CH2)24NRgRu, (CO)(CH2)24NReRy, (CO)NR(CH2)2NRgRx, :

   : (CO)O(CHa2)24NRGRH, SONRgRy, and (CO)NRgRy; Ce each of Reg, Rp, Rg, Rf, Ra, and Ry is independently selected from H, C 14 alkyl, So oo © C 34 alkenyl, C 34 alkynyl, C 55 cycloalkyl, and phenyl; each of NRcRop, NReRr, and NReR# can aiso be independently morphelinyl, piperazinyl, pyrrolidinyl, or piperadinyt; each of R, and R, is independently H, methyl, or ethyl; Rk is C 14 alkyl, C 54 alkenyl, C 14 alkynyl, C 35 cycloalkyl, or phenyl;

   : Xis QO, S, or NK; and E AMENDED SHEET

   PCT/US00/18348 wherein each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C , alkyl, C 3 cycloalkyl, C ,4 alkenyl, C 4 alkynyl, phenyl, hydroxyl, amino, (amino)sulfonyl, and NO,, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 2 substituents independently selected from halo, C ,, alkyl, hydroxyl, amino, and NO; or a pharmaceutically acceptable salt or C , ; ester thereof. :

   37. Use of claim 27, wherein R; is NO,.

   38. Use of claim 27, wherein R, is fluoro.

   39. Use of claim 27, wherein each of Ry; and Ris independently selected from H and fluoro. — : 40. Use of claim 27, wherein Rs is methyl, fluoro, or chloro.

   41. Use of claim 27, wherein R, is methyl, chloro, fluoro, nitro, or hydrogen.

   42. Use of claim 41, wherein Rg is H.

   43. Use of claim 41, wherein Rg is fluoro. 44, Use of claim 27, wherein R, is methyl or ethyl.

   45. Use of claim 27, wherein R, is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenyl, phenethyl, allyl, C ,5 alkenyl, C ,5 cycloalkyl, (C 3s cycloalkyl)C 4, alkyl, (C 55 heterocyclic radical)C ,_, alkyl, or (CH,),4 NRcRp. 127 "AMENDED SHEET

   ® | | PCT/US00/18348

   46. Use of claim 45, wherein R, is ; or (C ,, cycloalkyl)-C ,., alkyl. : 44, | Use of claim 27, wherein R, is H or methyl.

   48. Use of claim 27, wherein R, has at least one hydroxyl substituent.

   49. Use of claim 27, wherein R, is H, methyl, ethyl, isobutyl, hydroxyethyl, phenyl, 2-piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)- ) piperazin-1-yl]-propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino-ethyl; and Rg is H: or where Rj is methyl and R, is phenyl.

   50. Use of claim 27, wherein W is NR Rg or NR, NR, R;. . 51 . Use of claim 27, wherein W is NR,(CH,)>4 NR,Rg or O(CH,),5 NR4Rg.

   52. Use of claim 27, wherein W is NR,OR;. | oo

   53. Use of claim 27, wherein W is ORg. : . 54. Use of claim 27 wherein R, is in the para position relative to X.

   55. Use of claim 54, wherein R; is iodo.

   56. Use of claim i. wherein Rg is in the ortho position relative to X. 128 AMENDED SHEET

   PCT/US00/18348

   57. Use of claim 27 having the formula 2,4-bis-(2-chloro-4-iodo- phenylamino)-3-fluoro-5-nitro-benzoic acid.

   58. Use of claim 27, wherein said MEK inhibitor has a 1 structure selected from: 2-(2-chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-(4- sulfamoyl-phenylamino)-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-3- fluoro-S-nitro-4-phenylamino-benzoic ‘acid; 2-(2-chloro<4-iodo-phenyiamino)-3- fluoro-5-nitro-4-phenoxy-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-3- fluoro-5-nitro-4-phenyisulfanyl-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)- 3-fluoro-4-(methyl-phenyl-amino)-5-nitro-benzoic acid; benzamide, 2-[(2- chloro-4-iodophenyl)amino}-3-flucro-N-hydroxy—4-{[4-[[(2-hydroxyethyl)amino]- carbonyl]phenyllamino]-5-nitro-; benzamide, 2-[(2-chloro-4-iodophenyl)amino]- 4-[[4-[(dimethylamino)carbonyl]phenyilamino]-3-fluoro-N-hydroxy-5-nitro-; 2- (2-chloro-4-iodo-phenylamino)-3,5-diflucro-4-phenylamino-benzoic acid; 2-(2- chioro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-(3-sulfamoyl-phenylamino)- benzoic acid; and 2-(2-chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-4-(2- : sulfamoyl-phenylamino)-benzoic acid; and the comresponding hydroxamic acids and cyclopropylmethyl hydroxamates.

   59. Use of a MEK inhibitor selected from a compound of formula (DB: w-_-20 IST H ) N R4 ReRg J : ‘ Rs (he 129 AMENDED SHEET

   ° PCT/US00/18348 i wherein oo oo . | : Wis OR,, NR20OR;, NRaRs, NRzNRaRg, O(CH2):4NRaRa, or NRy{CHa)i4 ~ NRAaRg; - . O(CH2)14OR4, or NRz(CHgz)i4 ORy; : : oo © RyisH, Cigalkyl, Css alkenyl, C 35 alkynyl, C 3.5 cycloalkyl, phenyl, : (phenyl)C 14 alkyl, (phenyl)C 34 alkenyl, (phenyl)C 34 alkynyl, (C 38 cycloalkyl)- : EE C 14 alkyl, (C 15 cycloalkyl)C 14 alkenyl, (C 3g cycloalkyl)C 34 alkynyl, C 33 heterocyclic radical, (C 3s heterocyclic radical)C 14 alkyl, (C 38 heterocyclic radical)C a+ alkenyl, or (C 3s heterocyclic radical)C 34 alkynyl; oo : - .each of R> and Rj; is independently H, phenyl, C 14 alkyl, C 5g alkynyl, C3s ) oo | cycloalkyl, or (C 38 cycloalkyl)C 14 alkyl; each of Rs, Rs and Rg is independently H, CI, F, or Br; Ea - : Ce Ra is H, C 1 alkyl, C 35 alkenyl, C 35 alkynyl, C 35 cycloalkyl, phenyl, (C zs. | } : cycloalkyl)C 14 alkyl, (C 35 cycloalkyl)C 34 alkenyl, (C 33 cycloalkyl)C 34 CY alkynyt, C is heterocyclic radical, (C 28 heterocyclic radical)C 14 alkyl, (aminosulfonyl)phenyl, [(@minosulfonyl)phenyl]C 14 alkyl, (aminosulfonyl)C 16 alkyl, (amincsutfonyl)C sz cycloalkyl, or [(aminosulfonyl)C 35 cycloalkyi]C 14 alkyl; Co oo : Re is H, C 4.3 alkyl, C 35 alkenyl, C 15 alkynyl, C 3¢ cycloalkyl, or phenyl; Jis SRe, ORe, SOR, SOR, SO-NRpRE, Cc 1-8 alkyl, C 38 alkenyl, Cas . alkynyl, C ss cycloalkyl, C ss cycloalkenyl, phenyl, (C ss cycloalkyl)C 1+ alkyl, AMENDED SHEET 130 | Co

   PCT/US00/18348 (C 3.8 cycloalkyl)C 34 alkenyl, (C 3.4 cycloalkyl)C 34 alkynyl, C 33 heterocyclic radical, (C 34 heterocyclic radical)C 14 alkyl, -M'E'G’, (heterocyclic radical)- M’-E’-G’, or (cycloalkyl)-M'-E’-G’; M' is O, SO, SO, NRg, (CO)NRg, NRg (CO), SO;NRg, NReSO,, or

   CHa; .

   E’ is absent (a covalent bond), (CHa)14 or (CHz)m O(CH3), where 1 < (each of m and p independently) <3 and 2 <(m + p) < 4,

   G’ is OR3, SOzR¢, or NRgRg; provided that where p=1, then G'is H; : : | each of Rg, Rp, Re, Rr and Rg is independently selected from H, C 15 alkyl, C 34 alkenyl, C 34 alkynyl, C 15 cycloalkyl, C 15 heterocyclic radical,

   and phenyl; NReRg and NRpRe can each also independently be selected from

   ~ morpholinyl, pyrazinyl, piperazinyl, pyrrolidinyl, or piperadinyi; Rio is H, C 14 alkyl, halo, NO,, or SO;NRuR;; and R41 is H, halo, or NO; wherein each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 14 alkyl, C 35 cycloalkyl, C 24 alkenyl, C 24 alkynyl, phenyl, hydroxy,

   amino, (amino)sulfonyl, and NO,, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in tum optionally substituted with between 1 and 3 substituents independently selected from halo, C 4.z alkyl, hydroxy, amino, and NO; or a pharmaceutically acceptable salt or C ,_; ester thereof, in the manufacture of a medicament for treating chronic pain.

   131 AMENDED SHEET

   PCT/US00/18348

   60. Use of claim 59, wherein said chronic pain is selected from neuropathic : pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   61. . Use of claim 60, wherein said chronic pain is a type of neuropathic pain.

   62. Use of claim 61, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any : other nerve injury between the peripheral nervous system and the central nervous system, inclusively. 63 Use of claim 60, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism

   64. Use of claim 60, wherein said chronic pain is associated with idiopathic pain.

   65. Use of claim 59, wherein said chronic pain is associated with inflammation.

   66. Use of claim 59 wherein said chronic pain is associated with arthritis.

   67. Use of claim 59, wherein said chronic pain is associated with post-- operative pain.

   68. Use of claim 59, wherein R. is C ,, alkyl. 132 AMENDED SHEET

   ® | PCT/US00/18348

   69. Use of claim 59, wherein W is OH. : | : © 70. - Useof iim 59, wherein W is NHOH.

   71. Use of claim 59, wherein W is NHO(cyclopropyimethyl).

   72. Use of claim 59, wherein R,, is methyl or chloro.

   73. Use of claim 59, wherein R,, is fluoro.

   74. Use of claim 59, where Ry is H. :

   75. Use of claim 59, wherein J is trihalomethyl or methylthio.

   76. Use of claim 59, wherein J is + 2 5hiadiazol-3yl.

   77. Use of claim 59, wherein J is SO,CHa.

   78. Use of claim 59, wherein J is SOCHs,.

   79. Use of claim 59, wherein J is C ,4 alkynyl where the triple bond is between the carbon atoms alpha and beta to the phenyl group.

   80. Use of claim 59, wherein R, has at least one hydroxy substituent

   81. Use of claim 58, wherein R, is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 55 alkenyl, C 55 alkynyl, C 54 cycloalkyl,

   (C,.s cycloalky!)C ;, alkyl, or (C 55 heterocyclic radical)-C 1, alkyl. 133 : AMENDED SHEET

   ~ PCT/US00/18348

   82. Use of claim 59, wherein R, is H or (C 5, cycloalkyl)-C ,_, alkyl. :

   83. Use of claim 59, wherein R, is H, methyl, C 5, alkynyl, C 55 cycloalkyl, or (C45 cycloalkyl)methyl.

   84. Use of claim 59, wherein R, is H, methyl, ethyl, isobutyl, hydroxyethyl, ‘hydroxypropyl, cyclopropyimethyl, cyclobutylmethyl, C 5, alkynyl, phenyl, 2- piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-{4-(2-hydroxyethyl)-piperazin-1-yl}- propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino-ethyl; and Rg is H; orwhere Rg is methyl and R, is phenyl. :

   85. Use of claim 59, wherein each of R, and Rg is H, and Rs is F.

   86. Use of claim 59, wherein each of R,, Rand Rg is F.

   87. Use of claim 59, wherein each of R, and R; is F and Rg is Br.

   88. Use of claim 59, wherein Rs is F.

   89. Use of claim 59, wherein said MEK inhibitor has a structure selected from: 4-fluoro-2-(2-methyl-4-methylsufanyl-phenylamino)-benzoic acid; 5- bromo-3,4-difluoro-2-(2-methyl-4-methylsufanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid, 2-(4- methanesuifinyl-2-methyl-phenylamino)-4-nitro-benzoic acid; 3,4,5-trifluoro-2- (4-methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(2- methyl-4-methylsufanyl-phenylamino)-benzoic acid; 2-(2-methyl-4- - methylsulfanyl-phenylamino)-4-nitro-benzoic acid, 134 AMENDED SHEET

   PCT/US00/18348 C 3,4,5-trifluoro-2-(4-methanesuffinyl-2-methyl-phenylamino)-benzoic acid; 4- fluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid: 5-bromo-3 4- difluoro-2-(4-methanesulfonyl-2-methyi-phenylamino)-benzoic acid; 3,4,5- trifluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 4-fluoro-2-

   (4-methane-sulfinyl-2-methyl-phenylamino)-benzoic acid; 5-bromo-3,4- difluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 3,4- difiuoro-2-(4-methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 2-(4- methanesulfonyl-2-methyil-phenylamino)-4-nitro-benzoic acid; N-

   cyclopropylmethoxy-4-fluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)- . benzamide; 5-bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(2-methyi-4- . methylsulfanyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro- 2-(4-methanesulfinyl-2-methyl-phenylamino)-benzamide; N- cyclopropylmethoxy-2-(4-methanesuffinyl-2-methyl-phenylamino)-4-nitro- benzamide; N-cyclopropylmethoxy-3,4,5-trifluoro-2-(4-methanesulfonyl-2- methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(2- : : methyl-4-methylsulfanyl-phenylamino)-benzamide: N-cyclopropylmethoxy-2- (2-methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzamide; N- - cyclopropylmethoxy-3,4,5-trifluoro-2-(4-methanesulfinyi-2-methyl- phenylamino)-benzamide; N-cyclopropyimethoxy-4-fluoro-2-(4- © 20 methanesulfinyl-2-methyl-phenylamino)-benzamide; 5-bromo-N- : cyclopropylmethoxy-3,4-difluoro-2-(4-methanesutfonyl-2-methyl-

   : phenylamino)-benzamide; N-cyclopropyimethoxy-3,4,5-trifluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzamide; N-cyclopropylimethoxy-<4-fluoro-2-(4- methanesulfinyl-2-methyl-phenylamino}-benzamide; 5-bromo-N-

   ~ 25 cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-methanesuifonyi-2- methyl-phenylamino)-benzamide; and N-cyclopropyimethoxy-2-(4- methanesulfonyl-2-methyl-phenylamino)-4-nitro-benzamide. © 90. Use of claim 59 wherein said MEK inhibitor has a structure selected from: 4-fluoro-N-hydroxy-2-(2-methyl-4-methylsulfanyl- phenylamino)-benzamide; 5-bromo-3,4-difluoro-N-hydroxy-2-(2-methyl-4- 135 AMENDED SHEET

   ® PCT/US00/18348 methylsulfanyl-phenylamino)-benzamide; 3,4-difluoro-N-hydroxy-2-(4- methanesulfinyl-2-methyl-phenylamino)-benzamide; N-hydroxy-2-(4- ‘methanesulfinyl-2-methyl-phenylamino)-4-nitro-benzamide; 3,4,5-trifluoro-N- hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino)-benzamide; 3,4- difluoro-N-hydroxy-2-(2-methyl-4-methyisulfanyl-phenylamino)-benzamide; N- hydroxy-2-(2-methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzamide; 8:

   3.4 5-trifluoro-N-hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)- benzamide; 4-fluoro-N-hydroxy-2-(4-methanesuffinyl-2-methyl-phenylamino)- benzamide; 5-bromo-3,4-difluoro-N-hydroxy-2-(4-methanesulfonyl-2-methyi- phenylamino)-benzamide; 3,4,5-trifluoro-N-hydroxy-2-(2-methyl-4- methylsulfanyi-phenylamino)-benzamide; 4-fluoro-N-hydroxy-2-(4- methanesulfinyl-2-methyl-phenylamino)-benzamide; 5-bromo-3,4-difluoro-N- hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzamide; 3,4-difluoro- N-hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino)-benzamide: and N- hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino}-4-nitro-benzamide. -

   91. Use of claim 59, wherein said MEK inhibitor has a structure selected from: 3,4-difluoro-2-(4-imidazol-1-yl-2-methyl- phenylamino)-benzoic acid; N-cyciopropylmethoxy-3,4-difluoro-2-(4-imidazol- 1-yl-2-methyl-phenylamino)-benzamide; 3,4-difluoro-N-hydroxy-2-(4-imidazol- 1-yl-2-methyl-phenylamino)-benzamide; 3,4,5-trifluoro-2-(2-methyl4-

   [1.2,5]thiadiazol-3-yl-phenylamino)-benzoic acid; N-cyclopropyimethoxy-3,4,5- trifluoro-2-(2-methyl-4-[1,2,5]thiadiazol-3-yl-phenylamino)-benzamide; 3,4,5- trifluoro-N-hydroxy-2-(2-methyl-4-[1 ,2,5thiadiazol-3-yl-phenylamino)- benzamide; 2-[4-(4-chioro-[1,2,5}thiadiazol-3-yl)-2-methyl-phenylamino}-3,4,5- triftuoro-benzoic acid; 2-[4-(4-chloro-[1,2,5]thiadiazol-3-yl)-2-methyl- phenylamino]-N-cyclopropyimethoxy-3,4,5-trifluoro-benzamide; 2-{4-(4-chloro- [1,2,5]thiadiazol-3-yl)-2-methyl-phenylamino}-3,4, 5-trifluoro-N-hydroxy- benzamide; 2-{4-[4-(2-dimethylamino-ethoxy)-[1,2,5}thiadiazol-3-yl}-2-methyi- phenylamino}-3,4,5-trifluoro-benzoic acid; N-cyclepropylmethoxy-3,4,5- trifluoro-2-{2-methyl-4-[4-(2-piperidin-1 -yl-ethoxy)-{1,2,5}thiadiazol-3-yl}- 136 AMENDED SHEET

   ~ PCT/US00/18348 ® phenylamino}-benzamide; and 3,4, 5-trifluoro-N-hydroxy-2-{2-methyl-4-[4-(2- morpholin4-yl-ethoxy)-(1 2,5]thiadiazol-3-yl}-phenylamino}-benzamide.

   +92. Use of claim 59, wherein said MEK inhibitor has a - structure selected from: 5-bromo-2-(2-chloro<4-methylsulfanyl-phenylamino)-

   3.4-difluoro-benzoic acid; 2-(2-chloro-4-methanesulfinyl-phenytamino)-3,4- - difluoro-benzoic acid; 2-(2-chloro-4-methanesulfonyl-phenylamino)-3,4,5- trifluoro-benzoic acid; 2-(2-chloro-methylsulfanyl-phenylamino)-3,4-difluoro- benzoic acid; 5-bromo-2-(2-chloro-4-methanesulfonyl-phenylamino)-3,4- difluoro-benzoic acid; 2-(2-Chioro-4-methanesulfonyl-phenylamino)-3,4- difluoro-benzoic acid: 5-bromo-2-(2-chioro-4-methylsulfanyl-phenylamino)-N- cyclopropyimethoxy-3,4-difluoro-benzamide; 2-(2-chioro-4-methanesulfinyl- ~ phenylamino)-N-cyclopropylmethoxy-3,4-diflucro-benzamide; 2-(2-chloro—4- : methanesulfonyl-phenylamino)- N-cyclopropyimethoxy-3,4,5-trifluoro- benzamide,; 2-(2-chloro4-methylsulfanyl-phenylamino)- N- ) cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-methanesuifinyl-phenylamino)- N-cyclopropylmethoxy-3.4,5- : trifluoro-benzamide; 5-bromo-2-(2-chloro-4-methanesulfonyl-phenylamino)-N- : cyclopropyimethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-methylisulfanyi- phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-(2-chioro-4- methanesuifonyl-phenylamino)-N-cyclopropyimethoxy-3,4-diflucro-benzamide; 2-[2-chloro 4-(3H-imidazol-1-yl)-phenylamino}-N-cyclopropylmethoxy-3,4- difluoro-benzamide; 2-(2-chloro-4-[1,2,5]thiadiazol-3-yl-phenylamino)-N- cyclopropylmethoxy-3,4,5-triflucro-benzamide; 2-[4-(2-chioro4-chloro- [1,2,51thiadiazol-3-yl)-phenylamino]-3,4,5-trifluoro-benzoic acid; 2-[2-chloro4- (4-chloro-[1 ,2,5}thiadiazol-3-yl)-phenytamino]-N-cyclopropylmethoxy-3,4,5- trifluoro-benzamide; 2-{4-[4-(2-dimethylamino-ethoxy)-[1,2,5}thiadiazol-3-yi}-2- methyl-phenylamino}-3,4,5-trifluoro-benzoic acid; and 2-{2-chloro-4-[4-(2- piperidin-1-yl-ethoxy)-[1,2,5]thiadiazol-3-yl}-phenylamino}-N- cyclopropylmethoxy-3,4,5-trifluoro-benzamide. 137 AMENDED SHEET oo

   PCT/US00/18348 ®

   93. Use of claim 59, wherein said MEK inhibitor has a structure selected from: 2-(4-Ethynyl-2-methyi-phenylamino)-4-fluoro-benzoic’ acid; 5-Bromo-2-(4-ethynyi-2-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropyimethoxy-2-(4-ethynyi-2-methyl-phenylamino)-3,4-difluoro- benzamide; N-Cyclopropylmethoxy-2-(4-ethynyl-2-methyl-phenylamino)-4- nitro-Benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)-3,4,5-trifluoro-N- hydroxy-benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(4-Ethynyl-2-methyl-phenylamino)-4-nitro-benzoic acid; N- Cyclopropyimethoxy-2-(4-ethynyl-2-methyl-phenylamino)-3,4, 5-trifluoro- benzamide; 4-Fluoro-N-hydroxy-2-(4-methanesulfinyl-2-methyi-phenylamino)- benzamide; 5-Bromo-2-(4-ethynyl-2-methyl-phenylamino)-3,4-difluoro-N- hydroxy-benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)-3,4,5-trifluoro- benzoic acid; N-Cyclopropylmethoxy-2-(4-ethynyl-2-methyl-phenylamino)-4- fluoro-benzamide; 5-Bromo-N-cyclopropylmethoxy-2-(4-ethynyl-2-methyl- phenylamino)-3,4-difluoro-benzamide; 2-(4-Ethynyl-2-methyi-phenylamino)- : 3,4-diflucro-N-hydroxy-benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)-N- hydroxy-4-nitro-benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)-4-fluoro- benzoic acid; N-Cyclopropyimethoxy-2-(4-ethynyl-2-methyl-phenylamino)<4- ~ fluoro-benzamide; and 4-Fluoro-N-hydroxy-2-(4-methanesulfinyl-2-methyl- phenylamino)-benzamide.

   94. Use of claim 59, wherein said MEK inhibitor has a structure selected from: 2-(2-Chloro4-ethynyl-phenylamino)-4-fluoro-benzoic acid; 5-Bromo-2-(2-chloro4-ethynyi-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro4-ethynyl-phenylamino)- N-cyclopropylmethoxy-3.4-diflucro- benzamide; 2-(2-Chioro-4-ethynyl-phenylamino)- N-cyclopropyimethoxy-4- ’ nitro-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)- N-hydroxy-3,4,5- trifluoro- benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)-3,4-difluoro-benzoic acid: 2-(4-Ethynyl-2-chloro-phenylamino)-4-nitro-benzoic acid; 2-(2-Chloro-4- ethynyl-phenylamino)- N-Cyclopropyimethoxy-3,4,5-trifluoro-benzamide; 2-(2- chloro-4-methanesulfinyl-phenylamino)- 4-fluoro-N-hydroxy-benzamide; 5- Bromo-2-(4-ethynyl-2-chloro-phenylamino)-3,4-diflucro-N-hydroxy-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-Chloro- 138 AMENDED SHEET

   PCT/US00/18348 4-ethynyl-phenylamino)- N-cyclopropylmethoxy-4-fluoro-benzamide; 5-Bromo- : 2-(2-chloro-4-ethynyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(4-Ethynyl-2-chloro-phenylamino)-3,4-difluoro-N-hydroxy- benzamide; 2-(4-Ethynyl-2-chloro-phenylamino)-N-hydroxy-4-nitro-

   5 . benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)-4-fluoro-benzoic acid; 2-(2- Chloro-4-ethynyl-phenylamino)- N-cyclopropylmethoxy-4-fluoro-benzamide; 2- (2-Chloro4-methanesulfinyl-phenylamino)- 4-flucro-N-hydroxy-benzamide; and 2-(2-chloro4-imidazol-1-yl-phenylamino)- 3,4-Difluoro-benzoic acid.

   85. Use ofaMEK inhibitor selected from a compound of formula (1)C: w,__O - HRs di OL R4RNO-S—+ : me 2 NS - : A : (he wherein : Wis ORy, NR,ORjy, NRARg, NR.oNRARE, or NR(CH3)24 NRARE; ’ R1is H, C 13 alkyl, C 33 alkenyl, C 35 alkynyl, C 3.5 cycloalkyl, phenyl, (phenyi)C ,4 alkyl, (phenyi)C 14 alkenyl, (phenyl)C 34 alkynyl, (C 13 cycloalkyt)- : 139 AMENDED SHEET

   ® | | PCT/US00/18348 N C + alkyl, (C 54 cycloalkyi)C 34 alkenyl, (C ss cycloalkyl)C a4 alkynyl, C 5.3 heterocyclic radical, (C 3-8 heterocyclic radical)C 14 alkyl, (C 35 heterocyclic radical)C 14 alkenyl, (C zs heterocyclic radical)C 14 alkynyl or (CH,)24NRaRs; Rgis H, phenyl, C 14 alkyl, Cs4 alkenyi, Cc ss alkynyl, C 35 cycloalkyl, or - - © (C 35 cycloalkyl)-C 14 alkyl; : I Rais H, C 1 alkyl, C 35 alkenyl, C 3s alkynyl, C 33 cycloalkyl, phenyl, (C 38 Co ~~ cycloalkyl)C 14 alkyl, (C zg cycloalkyl)C 54 alkenyl, (C 3. cycloalkyl)C 34 . oo © 10 . alkynyl, C zg heterocyclic radical, (C 3s heterocyclic radical)C 1-4 alkyl, (aminosulfonyl)phenyl, [(aminosulfonyl)phenyl]C 14 alkyl, (aminosuifonyl)C 1s ST alkyl, (aminosulfonyl)C 35 cycloalkyl, or [(aminosulfonyl)C 35 cycloalkyi]C 14 } ~alkyi; FE RgisH,C 18 alkyl, C 35 alkenyl, C 28 alkynyl, C sg cycloalkyl, orC sg aryl; | | Co ‘Rs is H, F, Cl, Br, or NOg; | oo EE. ) Re is H or F; co LL oo Rs is H, methyl or Cl; Rs is H, C 14 alkyl, hydroxyethyl, hydroxypropyl, (CHz)2+(NRcRp), phenyl, 2- pyridyl, 3-pyridyl, 4-pyridyl or CH Ar, where Ar is phenyl, 2-pynidyl, 3-pyridyi, © or 4-pyridyi; R7 is H, C 14 alkyl, hydroxyethyl, hydroxypropyl, (CHa)2+(NRcRp), phenyl, 2- : pyridyl, 3-pyridyl, 4-pyridyl, or CHz2Ar , where Ar is phenyl, 2-pyridyl, 3-pyridyi, : or 4-pyndyl; : | oo each of Re and Rp is independently selected from H, C 15 alkyl, C 3 alkenyt, AMENDED SHEET :

   PCT/US00/18348 C a4 alkynyl, C 34 cycloalkyl, C 35 heterocyclic radical, and phenyl; NRcRp : can also be selected from morpholinyl, piperazinyl, pyrrolidinyl, or piperadinyl; wherein each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 14 alkyl, C 35 cycloalkyl, C ,4 alkenyl, C 54 alkynyl, phenyl, hydroxy, amino, (amino)sulfonyl, and NO,, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C 4.; alkyl, hydroxy, amino, and NO; - or a pharmaceutically acceptable salt or C , ; ester thereof E in the manufacture of a medicament for treating chronic pain.

   96. Use of claim 95, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism. :

   97. Use of claim 96, wherein said chronic pain is a type of neuropathic pain.

   98. Use of claim 97, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, | Co gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other nerve injury between the peripheral nervous system and the central nervous system, inclusively. 99 Use of claim 96, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism. 141 AMENDED SHEET

   PY PCT/US00/18348

   100. Use of claim 96, wherein said chronic pain is associated with idiopathic pain.

   101. Use of claim 95, wherein said chronic pain is associated with inflammation.

   102. Use of claim 95, wherein said chronic pain is associated with arthritis.

   103. Use of claim 95, wherein said chronic pain is associated with post- operative pain.

   104. Use of claim 95, wherein the sulfamoy! group is meta to W(CO)- and para to the bridging NH.

   105. Use of claim 95, wherein the sulfamoy! group is para to W(CO)- and meta to the bridging NH.

   106. Use of claim 95, wherein R, is fluoro.

   107. Use of claim 95, wherein R; is fluoro. : 108. Use of claim 95, wherein R; is H.

   109. Use of claim 95, wherein each W is OH.

   110. Use of claim 95, wherein W is NR,OR,.

   111. Use of claim 109, wherein each of R; and R, is fluoro. :

   112. Use of claim 95, wherein R, has at least one hydroxy substituent. 142 AMENDED SHEET

   ® PCT/US00/18348 ~ 113. Useof claim 95, wherein R, is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 55 alkenyl, C 55 alkynyl, C 34 cycloalkyl, (C45 cycloalkyl)C 4 alkyl, or (C 55 heterocyclic radical)-C 4; alkyl.

   114. Use of claim 113, wherein R, is H or (C 5, cycloalkyl)-C ,, alkyl.

   115. Use of claim 95, wherein R, is H, methyl, C 5, alkenyl, C ;5 cycloalkyl, or (C55 cycloalkyl)methyl.

   116. Use of claim 95, wherein R, is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C 54 alkynyl, phenyl, 2- : piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)-piperazin-1-yl]- propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino-ethyl; and Rg is H; or where Rg is methyl and R, is phenyl. Co © 117. Use of claim 95, wherein R; (CH,),4(NRcRp).

   118. Use of claim 95, wherein NR.R;, is selected from morpholinyl, piperazinyl, pyrrolidinyl, or piperadinyl.

   119. Use of claim 85, wherein Rg is methyl or chloro.

   120. Use of claim 95, wherein said MEK inhibitor has a structure selected from: 2-(2-chloro-4-iodo-phenylamino)-4-sulfamoyi-benzoic acid; 2- (2-chloro-4-iodo-phenylamino)-N-hydroxy-4-sulfamoyl-benzamide; 2-(2-chloro- 4-iodo-phenylamino)-N-cyclopropylmethoxy-4-sulfamoyl-benzamide; 2-(2- chloro-4-iodo-phenylamino)-4-(2-morpholin-4-yl-ethylsulfamoyl)-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-N-hydroxy-4-(2-morpholin-4-yl- ethylsulfamoyl)-benzamide; 2-(2-chioro-4-iodo-phenylamino)-N- 143 oo AMENDED SHEET

   ° PCT/US00/18348 cyclopropylmethoxy-4-(2-morpholin-4-yl-ethylsulfamoyl)-benzamide; 2-(2- chioro4-iodo-phenylamino)-3,4-difluoro-5-sulfamoyl-benzoic acid: 2-(2-chloro- 4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-5-sulfamoyl-benzamide; 2-(2-

   SE chioro-4-iodo-phenylamino)-N-cyclopropyimethoxy-3,4-difiuoro-5-sulfamoyl- - benzamide: 2-(2-chioro-4-iodo-phenylamino)-3,4-difluoro-5-(2-morpholin-4-yl- oC ~ ethylsulfamoyi)-benzoic acid; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N- hydroxy-5-(2-morpholin4-yl-ethyisulfamoyl)-benzamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difiuoro-5-(2-morpholin-4-yi- : : oo ethylsulfamoyl)-benzamide; 5-(bis-pyridin-3-yimethyl-sulfamoyl)-3,4-difluoro-2- (4-iodo-phenylamino)-benzoic acid; 5-(bis-pyridin-3-ylmethyl-sutfamoyl)-N- oo cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-benzamide: N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-(methyl-pyridin-3- -ylmethyl-suifamoyl)-benzamide; N-cyclopropyimethoxy-3,4-difluoro-2-(4-iodo- Co phenylamino)-5-[(pyridin-3-yimethyl)-sulfamoy-benzamide: N- | Co cyclopropylmethoxy-5-[(3-diethylamino-propyl)-pyridin-3-ylmethyl-sulfamoyl]- . 3 4-difluoro-2-(4-iodo-phenylamino)-benzamide: N-cyclopropylmethoxy-3,4- : difluoro-5-[(3-hydroxy-propyl}-pyridin-3-yimethyl-sulfamoyi]-2-(4-iodo- ) phenylamino)-benzamide; N-cyclopropylmethoxy-5-(ethyl-pyridin-3-yimethyl- oo N suifamoyl)-3,4difluoro-2-(4-iodo-phenylamino)-benzamide; N-

   cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-3-ylmethyl- sulfamoyl]-2-(4-iodo-phenylamino)-benzamide; 5-(bis-pyridin-2-ylmethyi- Co -sulfamoyl)-3,4-difluoro-2-(4-iodo-phenylamino)-benzoic acid; 5-(bis-pyridin-2- ylmethyl-sulfamoyl)-N-cyclopropyimethoxy-3,4-diflucro-2-(4-iodo-

   : phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- phenylamino)-5-(methyl-pyridin-2-ylmethyl-sulfamoyi)-benzamide; N- oo cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-{(pyridin-2- yimethyl)-sulfamoyl}-benzamide; 5-(bis-pyridin-3-ylmethyl-sulfamoyl)-3,4- : : diflucro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid; 5-(bis-pyridin-3- ylmethyl-sulfamoyl)-N-cyciopropyimethoxy-3,4-difiuoro-2-(4-iodo-2-methyl-

   phenylaminc)-benzamide; N-cyclopropylmethoxy-3,4-diftuoro-2-(4-iodo-2- methyl-phenylamino)-5-(methyl-pyridin-3-yimethyl-sutfamoyl)-benzamide; N-

   cyclopropyimethoxy-3 4-difiuoro-2-(4-iodo-2-methyl-phenylamino)-5-[(pyridin- Co AMENDED SHEET

   ® PCT/US00/18348 3-yimethyl)-sulfamoyl]-benzamide; N-cyclopr opylmethoxy-5-(3-diethylamino- propyl)-pyridin-3-ylmethyl-sulfamoyl]-3,4-difluoro-2-(4-iodo-2-methyl- | : : * phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difiuoro-5-{(3-hydroxy- propyl)-pyridin-3-ylmethyl-sulfamoyl}-2-(4-iodo-2-methyl-phenylamino)- benzamide; N-cyclopropylmethoxy-5-(ethyl-pyridin-3-yimethyl-sulfamoyf)-3,4- - difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy- 3 4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-3-ylmethyl-sulfamoyl]- 2-(4-iodo-2- : methyl-phenyiamino)-benzamide, 5-(bis-pyridin-2-ylmethyl-sulfamoyl)-3,4- difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid; 5-(bis-pyndin-2- | Co : 10 yimethyk-sulfamoyl)-N-cyclopropylmethoxy-3,4-difiuoro-2-(4-iodo-2-methyl- : 3 phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2- methyl-phenylamino)-5-(methyl-pyridin-2-yimethyk-sutfamoyl)-benzamide; N- oC cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{(pyridin- 2-ylmethyl)-sulfamoyli}-benzamide; 5-(bis-pyridin-3-yimethyl-sulfamoyl)-2-(2- - 15 chioro-4-iodo-phenylamino)-3,4-difluoro-benzoic acid: 5-(bis-pyridin-3- Co yimethyl-sulfamoy!)-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy- oo 3,4-difluoro-benzamide; 2-{2-chloro-4-iodo-phenyiamino)-N- : oo cyclopropyimethoxy-3,4-difiuoro-5-(methyl-pyridin-3-yimethyl-suffamoyl)- : oo benzamide; 2-(2-chloro4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4- 20° difluoro-5-[(pyridin-3-yimethyl)-sulfamoyl}-benzamide; 2-(2-chloro-4-iodo- phenylamino)}-N-cyclopropyimethoxy-5-{(3-diethylamino-propyl}-pyridin-3- yimethyl-sulfamoyi]-3,4-difluoro-benzamide; 2-(2-chloro-4-iodo-phenylamino)- N-cyclopropylmethoxy-3,4-difiuoro-5-[(3-hydroxy-propyl)-pyridin-3-ylmethy!- sulfamoyl]-benzamide; 2-(2-chloro-4-iodo-phenylamino)-N- : cyclopropyimethoxy-5-(ethyi-pyridin-3-yimethyl-sutfamoyi)-3,4-difluoro- benzamide, 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy-3,4- diflucro-5-[(2-hydroxy-ethyl)-pyridin-3-ylimethyl-suffamoyli}-benzamide; 5-{bis- oo pyridin-2-yimethyl-sutfamoyl)-2+2-chloro4-iodo-phenylamino)-3,4-diiiuoro- oo - benzoic acid; 5-(bis-pyridin-2-ylmethyl-sulfamoyi)-2~2-chioro-4-iodo- phenylamino)-N-cyclopropylmethoxy-3,4-difluorc-benzamide; 2-(2-chloro-4- iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difiuoro-5{methyl-pyridin-2- : yimethyi-sufamoyi)-benzamide; 2-(2-chloro4-iodo-phenylamino)-N- AMENDED SHEET

   ® PCT/USOO/I8348" + a cyciopropyimethoxy-3,4-difluoro-5-{(pyridin-2-yimethyl)-sulfamoyl}-be nzamide;

   - . N-cyclopropyimethoxy-3,4-difluore-5-[(3-hydroxy-propyl)-pyridin-2-yimethyl- } sulifamoyl]-2-(4-iodo-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-. Co difluoro-5-[(2-hydroxy-ethyl)-pyridin-2-yimethyl-sulfamoyl}-2-(4-iodo- | Co phenylamino)-benzamide; 5-(benzyl-pyridin-2-yimethyl-sulfamoyl)-N- ~~ cyclopropylmethoxy-3,4-difiuoro-2-(4-iodo-phenylamino)-benzamide; N- .. . : | cyclopropylmethoxy-3,4-diflucro-2-(4-iodo-phenylamino)-5-[(pyridin4- EE ylmethyl)-sulfamoy(}-benzamide; N-cyclopropylmethoxy-5-(ethyl-pyridin-4- yimethyl-sulfamoyl)-3,4-diftuoro-2-(4-iodo-phenylamino)-benzamide; N- cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-(methyl-pyridin-4- : ylimethyl-sulfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-5-[(3- : hydroxy-propyh)-pyridin-4-ylmethyl-sulfamoyl}-2-(4-iodo-phenylamino)- oo benzamide; N-cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-4- : - yimethyl-sulfamoyl}-2-(4-iodo-phenylamino)-benzamide; N- : oo cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-phenylamino)-5-(methyl-phenyl- Co sulfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- | Co phenylamino)-5-phenyisulfamoyl-benzamide; N-cyclopropylmethoxy-3.4- - Co oo difluoro-2-(4-iodo-phenylamino)-5-(pyridin-3-ylsulfamoyl)-benzamide; N- : ~ ‘cyclopropylmethoxy-3 A-difluoro-5-[(3-hyd roxy-propyl)-pyridin-2-ylmethyl- . 20 sulfamoyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide: N- oo cyclopropylmethoxy-3,4-difluoro-5-[(2-hyd roxy-ethyl)-pyridin-2-ylmethyl- = :

   EE. sulfamoyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide; 5-(benzyl-pyridin-2- - ylmethyl-sulfamoyl)-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl- Co phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2- : . 25 methyl-phenylamino)-5-{(pyridin-4-yimethyl)-sulfamoyl}-benzamide; N- . - cyclopropylmethoxy-5-(ethyl-pyridin-4-ylmethyl-sulfamoyl)-3 4-difluoro-2-(4- iodo-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2- (4-iodo-2-methyl-phenylamino)-5-(methyl-pyridin-4-ylmethyl-sulfamoyl)- benzamide, N-cyciopropylmethoxy-3,4-difluoro-5-{(3-hydroxy-propyl)-pyridin- } 4-yimethyl-sulfamoyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide; N- : cyclopropyimethoxy-3,4-difluoro-5-[(2-hydroxy-ethyl)-pyridin-4-vimsthy}- sulfamoyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide; N- : : 146 . Co : : AMENDED SHEET

   ° . PCT/USO0/18348 Co cyclopropylmethoxy-3,4-difiuoro-2-(4-iodo-2-methyl-phenylamino)-5-{methyl- h oo phenyl-sulfamoyl)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2- i methyl-phenylamino)-5-phenylsujfamoyl-benzamide; N-cyclopropylmethoxy- - Co 3,4-difluoro-2-(4-iod o-2-methyl-phenylamino)-5-(pyridin-3-ylsulfamoyl)- . - . : 5 benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropyimethoxy-3,4- oo : ; difluoro-5-{(3-hyd roxy-propyl)-pyridin-2-ylmethyl-sulfamoyl]l-benzamide; 2-(2- chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difiuoro-5-{(2-hydroxy- R ethyl)-pyridin-2-yimethyl-suifamoyi}-benzamide: 5-(benzyl-pyridin-2-yimethyi- sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4~ - : : 10- - difluoro-benzamide; 2-(2-chioro-4-iodo-phenylamino)-N-cyclopropyimethoxy- + 3,4-difluoro-5-(pyridin4-yimethyl)-suifamoyl}-benzamide; 2-(2-chloro-4-iodo- : : phenyiamino)-N-cyclopropylmethoxy-5-(ethyl-pyridin-4-yimethyl-sulfamoyl)- 3,4-difluoro-benzamide; 2-(2-chloro-4-iodo-phenyiamino)-N- . = oo BE cyclopropylmethoxy-3,4-difiuoro-5-(methyl-pyridin-4-ylmethyl-suliamoyl)- oo oo 15 ~benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4- = Co difluoro-5-{(3-hydroxy-propyl)-pyridin-4-ylmethyl-sulfamoyl]-benzamide; 2-(2- - chiofo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-[(2-hydroxy- ethy!)-pyridin-4-ylmethyl-sulfamoyl}-benzamide; 2-(2-chloro-4-icdo- - phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-(methyl-phenyl-sulfarmoyl)- 20: benzamide; 2-(2-chioro-4-iodo-phenylamino)-N-cyciopropylmethoxy-3,4- A : difluoro-5-phenylsulfamoyk-benzamide; 2-(2-chioro-4-iodo-phenyiamino)-N- : cyclopropylmethoxy-3,4-difiucro-5-(pyridin-3-ylsuifamoyl)-benzamide; N- cyclopropylmethoxy-2-(4-iodo-phenylamino)<4-phenylsulfamoyl-benzamide: - N-cyclopropyimethoxy-2-(4-iod o-phenylamino)-4-(pyridin-3-ylsulfamoyl)- E benzamide; N-cyclopropylmethoxy-2-(4-iodo-phenylamino)-4-{(pyridin-3- 'yimethyl)-sulfamoyl}-benzamide; 4-(bis-pyridin-3-ylmethyl-sulfamoyl)-N- | i oo cyclopropylmethoxy-2-(4-iodo-phenylamino)-benzamide; N- cyclopropylmathoxy-4-{(2-hydroxy-ethyl)-pyridin-4-yimethyl-sutfamoyJ-2-(4- oo | iodo-phenylamino)-benzamide; N-cyclopropylmethoxy-2-(4-iodo- : : phenylamino)-4-(methyl-pyridin-3-ylmethyl-sulfamoyl)-benzamide; N- cyclopropyimethoxy-4-{(3-diethylamino-propyl)-pyridin-3-yimethyl-sulfamoyl}- 2-(4-iodo-phenylamino)-benzamide; N-cyclopropylmethoxy-2-(4-iodo-2- : 147 AMENDED SHEET :

   * PCT/USH0/18348 methyl-phenylamino)-4-phenylsulfamoyl-benzamide; N-cyclopropylmethoxy-2- (4-iodo-2-methyl-phenylamino)-4-(pyridin-3-yisulifamoyl)-benzamide: N- cyclopropyimethoxy-2-(4-iodo-2-methyl-phenylamino)-4-[(pyridin-3-ylmethyl)- sulfamoyl]-benzamide; 4-(bis-pyridin-3-ylmethyl-sulfamoyl)-N- cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-benzamide; N- cyclopropylmethoxy-4-[(2-hydroxy-ethyl)-pyridin-4-yimethyl-sulfamoyl}-2-(4- iodo-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-2-(4-iodo-2- methyi-phenylamino)-4-(methyl-pyridin-3-ylmethyl-sulfamoyl)-benzamide; N- cyclopropylmethoxy-4-{(3-diethylamino-propyl)-pyridin-3-yimethyl-sulfamoyi}- : | 2-(4-iodo-2-methyil-phenylamino)-benzamide; 2-(2-chloro-4-iodo- ) oo phenylamino)-N-cyclopropyimethoxy-4-phenylsulfamoyl-benzamide: 2-(2- chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-(pyridin-3-ylsuffamoyl)- benzamide,; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4- : [(pyridin-3-ylmethyi)-sulfamoyil-benzamide; 4-(bis-pyridin-3-yimethyl- ‘ + sulfamoyl)-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy- benzamide,; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy—4-{(2- hydroxy-ethyt)-pyridin-4-yimethyl-sulfamoyi}-benzamide; 2-(2-chloro-4-iodo- phenylamino)-N-cyclopropylmethoxy-4-(methyl-pyridin-3-yimethyl-sulfamoyl)- benzamide; 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-{(3- diethylamino-propyl)-pyridin-3-yimethyl-suifamoyl]-benzamide; and 5-[bis-(4- methoxy-benzyl)-sulfamoyl}-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro- : benzoic acid; and 2-(2-chloro-4-iodo-phenylamino)-5-dimethylisuifamoyi-3,4- ) difluoro-benzoic acid methyl ester. :

   121. Use of claim 95, wherein said MEK inhibitor has a structure selected from: N-Allyloxy-2-(2-chloro-4-iodo- : phenylamino)-3,4-difluoro-5-(4-methyl-piperazine-1-sulfonyl)-benzamide; N-Allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5-(methyl- phenyl-sulfamoyl)-benzamide; 5-(Allyl-methyl-sulfamoyl)-N- allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzamide; 1-[5-Allyloxycarbamoyl-4-(2-chloro-4-iodo-phenylamino)-2,3-difluoro- : benzenesulfonyl]-piperidine-3-carboxylic acid amide; N-Allyloxy- 148 AMENDED SHEET

   PCT/US00/18348 2-(2-chloro-4-iodo-phenylamino)-5-[(3-dimethylamino-propyt)-methyl- sulfamoyl]-3,4-difluoro-benzamide; N-Allyloxy-2-(2-chloro-4-iodo- phenylamino)-3,4-difluoro-5-(4-pyridin-2-yl-piperazine-1-sulfonyl)-benzamide; : and N-Allyloxy-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-5- (methoxy-methyl-sulfamoyl)-benzamide.

   122. Use of claim 1, wherein said MEK inhibitor has a structure selected from: 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzenesulfonamide; and 2-(2-chloro-4-iodo-phenylamino)- * cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide. :

   123. Use of claim 27, wherein said MEK inhibitor has a structure selected from : 2,4-bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-benzoic acid.

   124. Use of claim 59, wherein said MEK inhibitor has a structure selected from : 2-(4-ethynyl-2-methyl-phenylamino)-4-fluoro-benzoic acid; and 2-(3',5'- dichloro-biphenyl-4-ylamino)-benzoic acid.

   125. Use of claim 95, wherein said MEK inhibitor has a structure selected from: 2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5- sulfamoyl-benzamide; 2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy- oo 5-sulfamoyl-benzamide; C-(2-chloro-4-iodo-phenylamino)-N- cyciopropylmethoxy-dimethylsulfamoyl-difluoro-benzamide; N- cyclopropylmethoxy-dimethylsulfamoyl-difluoro-C-(4-iodo--2-methyl- phenylamino)-benzamide; and C-(2-chloro-4-iodo-phenylamino)-difiuoro- (methoxy-methyl-sulfamoyl)-N-(2-morpholin-4-yl-ethoxy)benzamide.

   126. A substance or composition for use in a method for treating chronic pain, said substance or composition comprising a MEK inhibitor selected from : a compound of formula (I) as defined in claim 1, or a pharmaceutically acceptable salt or C , 5 ester thereof, and said method comprising administering said substance or composition to a subject in need of such treatment. 149 AMENDED SHEET

   PCT/US00/18348

   127. A substance or composition for use in a method of treatment according to claim 126, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   128. A substance or composition for use in a method of treatment according to claim 127, wherein said chronic pain is a type of neuropathic pain. :

   129. A substance or composition for use in-a method of treatment according to claim 128, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other nerve injury between the peripheral nervous system and the central nervous system, inclusively.

   130. A substance or composition for use in a method of treatment according to claim 127, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism

   131. A substance or composition for use in a method of treatment according to claim 127, wherein said chronic pain is associated with idiopathic pain.

   132. A substance or composition for use in a method of treatment according to claim 126, wherein said chronic pain is associated with inflammation. © 133. A substance or composition for use in a method of treatment according to claim 126 wherein said chronic pain is associated with arthritis.

   134. A substance or composition for use in a method of treatment according to claim 126, wherein said chronic pain is associated with post-operative pain. 150 AMENDED SHEET

   PCT/US00/18348

   135. A substance or composition for use in a method of treatment according to claim 126, wherein R; is bromo or chloro.

   136. A substance or composition for use in a method of treatment according to claim 126, wherein R, is fluoro.

   137. A substance or composition for use in a method of treatment according to claim 126, wherein R; is H.

   138. A substance or composition for use in a method of treatment according to claim 137, wherein each of R, and R; is H.

   139. A substance or composition for use in a method of treatment according to claim 126, wherein each of R, and Rj is fluoro.

   140. A substance or composition for use in a method of treatment according to claim 139, wherein R, is bromo.

   141. A substance or composition for use in a method of treatment according to claim 139, wherein R; is fluoro.

   142. A substance or composition for use in a method of treatment according } to claim 126, wherein R, is nitro.

   143. A substance or composition for use in a method of treatment according to claim 141, wherein R; is H. :

   144. A substance or composition for use in a method of treatment according to claim 126, wherein Ry is chloro.

   145. A substance or composition for use in a method of treatment according to claim 126, wherein R; is methyl. 151 AMENDED SHEET

   : PCT/US00/18348

   146. A substance or compasition for use in a method of treatment according to claim 126, wherein R,is Hor C ,, alkyl, and R, is H.

   147. A substance or composition for use in a method of treatment according to claim 126, wherein R, is (C ; 4 cycloalkyl) methyl.

   148. A substance or composition for use in a method of treatment according to claim 126, wherein R, is H.

   149. A substance or composition for use in a method of treatment according to claim 126, wherein R, is (CH,),, OR; or (CH,) ,4 NRcR. :

   150. A substance or composition for use in a method of treatment according to claim 126, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 25.

   151. A substance or composition for use in a method of treatment according ] to claim 126, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 26.

   152. A substance or composition for use in a method for treating chronic pain, said substance or compasition comprising a MEK inhibitor selected from a compound having the formula (l)A as defined in claim 27, or a oo pharmaceutically acceptable salt or C ,; ester thereof, said substance or composition comprising administering said substance or composition to a subject in need of such treatment.

   153. A substance or composition for use in a method of treatment according to claim 152, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism. 152 AMENDED SHEET

   PCT/US(0/18348

   154. A substance or composition for use in a method of treatment according to claim 153, wherein said chronic pain is a type of neuropathic pain.

   155. A substance or composition for use in a method of treatment according to claim 154, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other nerve injury between the peripheral nervous system and the central nervous system, inclusively.

   156. A substance or composition for use in a method of treatment according : to claim 153, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   157. A substance or composition for use in a method of treatment according to claim 153, wherein said chronic pain is associated with idiopathic pain.

   158. A substance or composition for use in a method of treatment according to claim 152, wherein said chronic pain is associated with inflammation.

   159. A substance or composition for use in a method of treatment according to claim 152 wherein said chronic pain is associated with arthritis.

   160. A substance or composition for use in a method of treatment according to claim 152, wherein said chronic pain is associated with post-operative pain. 153 AMENDED SHEET :

   PCT/US00/18348

   161. A substance or composition for use in a method of treatment according to claim 152, wherein the MEK inhibitor has the formula (1)A as defined in claim 36, or a pharmaceutically acceptable salt or C ,, ester thereof. } 162. A substance or composition for use in a method of treatment according to claim152, wherein R; is NO,.

   163. A substance or composition for use in a method of treatment according to claim 152, wherein R, is fluoro.

   164. A substance or composition for use in a method of treatment according : to claim 152, wherein each of R; and R, is independently selected from H and fluoro.

   165. A substance or composition for use in a method of treatment according to claim 152, wherein R; is methyl, fluoro, or chloro.

   166. A substance or composition for use in a method of treatment according to claim 152, wherein Ry is methyl, chloro, fluoro, nitro, or hydrogen.

   167. A substance or composition for use in a method of treatment according to claim 166, wherein R; is H.

   168. A substance or composition for use in a method of treatment according to claim 166, wherein Rs is fluoro. :

   169. A substance or composition for use in a method of treatment according to claim 152, wherein Ry is methyl or ethyl. 154 AMENDED SHEET

   PCT/US00/18348 ’

   170. A substance or composition for use in a method of treatment according to claim 152, wherein R; is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenyl, phenethyl, allyl, C , 5 alkenyl, C , ¢ cycloalkyl, (C 5 cycloalkyl)C ,, alkyl, (C 55 heterocyclic radical)C ,_, alkyl, or (CH,),, NR:Rp.

   171. A substance or composition for use in a method of treatment according : to claim 170, wherein R, is H or (C ,, cycloalkyl)-C ,, alkyl.

   172. A substance or composition for use in a method of treatment according to claim 152, wherein R, is H or methyl. ~ 173. A substance or composition for use in a method of treatment according to claim 152, wherein R, has at least one hydroxy! substituent.

   174. A substance or composition for use in a method of treatment according to claim 152, wherein R, is H, methyl, ethyl, isobutyl, hydroxyethyl, phenyl, 2- piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)-piperazin-1-yl]- propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino-ethyl; and Rg is H; or where Rg . is methyl and R, is phenyl. )

   175. A substance or composition for use in a method of treatment according to claim 152, wherein W is NR,R; or NR,NR,Rg.

   176. A substance or composition for use in a method of treatment according to claim 152, wherein W is NR,(CH,),, NR,Rg or O(CH,), ; NR,R;.

   177. A substance or composition for use in a method of treatment according to claim 152, wherein W is NR,OR,.

   178. A substance or composition for use in a method of treatment according to claim 152, wherein W is OR;. 155 AMENDED SHEET

   PCT/US00/18348

   179. A substance or composition for use in a method of treatment according to claim 152, wherein R, is in the para position relative to X.

   180. A substance or composition for use in a method of treatment according to claim 179, wherein R; is iodo. :

   181. A substance or composition for use in a method of treatment according to claim 152, wherein R; is in the ortho position relative to X.

   182. A substance or composition for use in a method of treatment according to claim 152, having the formula 2,4-bis-(2-chloro-4-iodo-phenylamino)-3- fluoro-5-nitro-benzoic acid. -

   183. A substance or composition for use in a method of treatment according to claim 152, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 58.

   184. A substance or composition for use in a method for treating chronic pain, said substance or composition comprising a MEK inhibitor selected from a compound of formula (1)B as defined in claim 59, or a pharmaceutically acceptable salt or C ,, ester thereof, said method comprising administering said substance or composition to a subject in need of such treatment.

   185. A substance or composition for use in a method of treatment according to claim 184, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   186. A substance or composition for use in a method of treatment according to claim 185, wherein said chronic pain is a type of neuropathic pain. 156 AMENDED SHEET

   PCT/US00/18348

   187. A substance or composition for use in a method of treatment according to claim 186, wherein said neuropathic pain is associated with one of the : following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other - nerve injury between the peripheral nervous system and the central nervous system, inclusively. . 188 A substance or composition for use in a method of treatment according to claim 185, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism

   189. A substance or composition for use in a method of treatment according to claim 185, wherein said chronic pain is associated with idiopathic pain.

   190. A substance or composition for use in a method of treatment according to claim 184, wherein said chronic pain is associated with inflammation.

   191. A substance or composition for use in a method of treatment according to claim 184, wherein said chronic pain is associated with arthritis.

   192. A substance or composition for use in a method of treatment according to claim 184, wherein said chronic pain is associated with post-operative pain.

   193. A substance or composition for use in a method of treatment according to claim 184, wherein R. is C ,, alkyl.

   194. A substance or composition for use in a method of treatment according to claim 184, wherein W is OH. 157 AMENDED SHEET

   PCT/US00/18348

   185. A substance or composition for use in a method of treatment according to claim 184, wherein W is NHOH.

   196. A substance or composition for use in a method of treatment according to claim 184, wherein W is NHO(cyclopropylmethyl).

   197. A substance or composition for use in a method of treatment according to claim 184, wherein R,, is methyl or chloro.

   198. A substance or composition for use in a method of treatment according to claim 184, wherein Ry, is fluoro.

   199. A substance or composition for use in a method of treatment according to claim 184, wherein R,; is H.

   200. A substance or composition for use in a method of treatment according to claim 184, wherein J is trihalomethyl or methylthio. oo

   201. A substance or composition for use in a method of treatment according to claim 184, wherein J is 1,2,5-thiadiazol-3-yl.

   202. A substance or composition for use in a method of treatment according to claim 184, wherein J is SO,CH;.

   203. A substance or composition for use in a method of treatment according to claim 184, wherein J is SOCH,.

   204. A substance or composition for use in a method of treatment according to claim 184, wherein J is C ,; alkynyl where the triple bond is between the carbon atoms alpha and beta to the phenyl group. 158 : AMENDED SHEET

   PCT/US00/18348

   205. A substance or composition for use in a method of treatment according to claim 184, wherein R, has at least one hydroxy substituent.

   206. A substance or composition for use in a method of treatment according to claim 184, wherein R, is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 55 alkenyl, C ;5 alkynyl, C ,4 cycloalkyl, (C; 5 cycloalkyl)C ,., alkyl, or (C ,_5 heterocyclic radical)-C ,_, alkyl. : :

   207. A substance or composition for use in a method of treatment according to claim 184, wherein R, is H or (C , cycloalkyl)-C ,_, alkyl.

   208. A substance or composition for use in a method of treatment according to claim 184, wherein R, is H, methyl, C ,, alkynyl, C , 5 cycloalkyl, or (C4 cycloalkyl)methyl.

   209. A substance or composition for use in a method of treatment according to claim 184, wherein R, is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C ,, alkynyl, phenyl, 2- piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)-piperazin-1-yl]- propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino-ethyl; and R; is H; or where Rg : is methyl and R, is phenyl. oo

   210. A substance or composition for use in a method of treatment according to claim 184, wherein each of R, and Rg is H, and Rg is F.

   211. A substance or composition for use in a method of treatment according to claim 184, wherein each of R,, R;and Ris F.

   212. A substance or composition for use in a method of treatment according to claim 184, wherein each of R, and Rs is F and R; is Br. 159 AMENDED SHEET

   PCT/US00/18348

   213. A substance or composition for use in a-method of treatment according to claim 184, wherein Ry is F.

   214. A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 8S.

   215. A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 90. co 216. - A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 91.

   217. A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 92.

   218. A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from the compounds listed in claim.93. : : 219. A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 94.

   220. A substance or composition for use in a method for treating chronic : pain, said method comprising a MEK inhibitor selected from a compound of ~ formula (1)C as defined in claim 95, or a pharmaceutically acceptable salt or

   : . C,, ester thereof, said method comprising administering said substance or composition to a subject in need of such treatment. 160 AMENDED SHEET

   PCT/US00/18348

   221. A substance or composition for use in a method of treatment according to claim 220, wherein said chronic pain is selected from neuropathic pain, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism.

   222. A substance or composition for use in a method of treatment according to claim 221, wherein said chronic pain is a type of neuropathic pain.

   223. A substance or composition for use in a method of treatment according to claim 222, wherein said neuropathic pain is associated with one of the following: inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and any other nerve injury between the peripheral nervous system and the central nervous system, inclusively. 224 A substance or composition for use in a method of treatment according to claim 221, wherein said chronic pain is associated with chronic alcoholism, vitamin deficiency, uremia, or hypothyroidism

   225. A substance or composition for use in a method of treatment according to claim 221, wherein said chronic pain is associated with idiopathic pain.

   226. A substance or composition for use in a method of treatment according to claim 220, wherein said chronic pain is associated with inflammation.

   227. A substance or composition for use in a method of treatment according to claim 220, wherein said chronic pain is associated with arthritis. 161 AMENDED SHEET

   PCT/US00/18348

   228. A substance or composition for use in a method of treatment according to claim 220, wherein said chronic pain is associated with post-operative pain.

   229. A substance or composition for use in a method of treatment according to claim 220, wherein the sulfamoyl group is meta to W(CO)- and para to the bridging NH.

   230. A substance or composition for use in a method of treatment according "to claim 220, wherein the sulfamoyl group is para to W(CO)- and meta to the bridging NH.

   231. A substance or composition for use in a method of treatment according to claim 220, wherein R, is fluoro.

   232. A substance or composition for use in a method of treatment according to claim 220, wherein R; is fluoro. . 233. A substance or composition for use in a method of treatment according to claim 220, wherein R; is H. oo

   234. A substance or composition for use in a method of treatment according to claim 220, wherein each W is OH.

   235. A substance or composition for use in a method of treatment according to claim 220, wherein W is NR,OR;.

   236. A substance or composition for use in a method of treatment according to claim 234, wherein each of R; and R, is fluoro.

   237. A substance or composition for use in a method of treatment according to claim 220, wherein R, has at least one hydroxy substituent. 162 AMENDED SHEET

   PCT/US00/18348

   238. A substance or composition for use in a method of treatment according : to claim 220, wherein R, is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 55 alkenyl, C alkynyl, C ;4 cycloalkyl, (C, 5 cycloalkyl)C ,, alkyl, or (C ;5 heterocyclic radical)-C ,_, alkyl.

   239. A substance or composition for use in a method of treatment according to claim 238, wherein R, is H or (C ,, cycloalkyl)-C ,_, alkyl.

   240. A substance or composition for use in a method of treatment according to claim 220, wherein R, is H, methyl, C ,, alkynyl, C ; ; cycloalkyl, or (Cis oo cycloalkyl)methyl.

   241. A substance or composition for use in a method of treatment according to claim 220, wherein R, is H, methyl, ethyl, isobutyl, hydroxyethyl, : hydroxypropyl, cyclopropyimethyl, cyclobutylmethytl, C ,, alkynyl, phenyl, 2- piperidin-1-yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)-piperazin-1-yl]- propyl, 2-pyrrolidin-1-yl-ethyl, or 2-diethylamino-ethy!; and Ry is H; or where Rg : is methyl and R, is phenyl.

   242. A substance or composition for use in a method of treatment according to claim 220, wherein R; (CH,),,(NR.Rp).

   243. A substance or composition for use in a method of treatment according to claim 220, wherein NR_R;, is selected from morpholinyl, piperazinyl, pyrrolidinyl, or piperadinyl.

   244. A substance or composition for use in a method of treatment according to claim 220, wherein R; is methyl or chloro.

   245. A substance or composition for use in a method of treatment according to claim 220, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 120. 163 AMENDED SHEET

   ® PCT/US00/18348

   246. A substance or composition for use in a method of treatment according to claim 220, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 121.

   247. A substance or composition for use in a method of treatment according to claim 126, wherein said MEK inhibitor has a structure selected from: 2-(2- chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzenesulfonamide; and 2-(2-chloro-4-iodo-phenylamino)- cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide.

   248. A substance or composition for use in a method of treatment according to claim 152, wherein said MEK inhibitor has a structure selected from : 2,4- bis-(2-chloro-4-iodo-phenylamino)-3-fluoro-5-nitro-benzoic acid.

   249. A substance or composition for use in a method of treatment according to claim 184, wherein said MEK inhibitor has a structure selected from : 2-(4- ethynyl-2-methyl-phenylamino)-4-fluoro-benzoic acid; and 2-(3',5'-dichloro- : biphenyl-4-ylamino)-benzoic acid.

   250. A substance or composition for use in a method of treatment according to claim 220, wherein said MEK inhibitor has a structure selected from the compounds listed in claim 125.

   251. Use according to any one of claims 1 - 125, substantially as herein described and illustrated.

   252. A substance or composition for use in a method of treatment according to any one of claims 126 - 250, substantially as herein described and illustrated. 164 AMENDED SHEET

   ® PCT/US00/18348.

   253. A new use of a compound which is defined or listed in any one of claims 1 - 125, or a substance or composition for a new use in a method of treatment, substantially as herein described. 165 AMENDED SHEET