HK1193032B

HK1193032B - Derivatives of n-(arylamino) sulfonamides including polymorphs as inhibitors of mek as well as compositions, methods of use and methods for preparing the same

Info

Publication number: HK1193032B
Application number: HK14106509.5A
Authority: HK
Inventors: J-M．韦尼耶; C．E．罗林斯; J-L．吉拉德特; S．迪莫克; B．夸尔特; J．N.．迈纳
Original assignee: Ardea Biosciences, Inc.
Priority date: 2007-07-30
Filing date: 2014-06-27
Publication date: 2017-10-20

Description

N- (arylamino) sulfonamide derivatives and compositions, including polymorphs, as MEK inhibitors, methods of use, and methods of preparation thereof

The present application is a divisional application of the chinese patent application having application number 200880108829.6, filing date 28/7/2008 entitled "N- (arylamino) sulfonamide derivatives and compositions, including polymorphs, as MEK inhibitors, methods of use and methods of making the same".

Cross Reference to Related Applications

Priority of U.S. provisional application No. 61/044,886 filed on 14.4.2008, U.S. provisional application No. 61/034,466 filed on 6.3.2008, and U.S. provisional application No. 61/034,464 filed on 6.3.2008, each of which is incorporated herein by reference in its entirety. This application also claims priority from U.S. application No. 11/830,733 filed on 30/7/2007, which claims benefit from U.S. provisional application No. 60/833,886 filed on 28/7/2006 and benefit from international application No. PCT/US2006/028326 filed on 21/7/2006 as a partial continuation, each of which is incorporated herein by reference in its entirety. International application No. PCT/US2006/028326 also claims priority from US provisional application No. 60/701,814 filed on 21/7/2005; the priority of 60/706,719 filed on 8/2005 and the priority of 60/731,633 filed on 28/2005, each of which is incorporated herein by reference in its entirety.

Technical Field

The present invention relates to N- (2-arylamino) arylsulfonamide compounds that are inhibitors of MEK including crystalline polymorphs that exhibit a specific powder X-ray diffraction pattern and/or a specific differential scanning calorimetry pattern. The invention also relates to pharmaceutical compositions comprising the compounds described herein and methods of using the compounds and compositions described herein, including use in the treatment and/or prevention of cancer, hyperproliferative diseases, and inflammatory conditions. The invention also relates to methods of making the compounds and compositions described herein.

Background

Oncogenes-genes that cause the production of cancer-are often mutated forms of certain normal cellular genes ("proto-oncogenes"). Oncogenes typically encode aberrant forms of signaling pathway components (e.g., receptor tyrosine kinases, serine-threonine kinases, or downstream signaling molecules). The major downstream signaling molecule is the Ras protein, which is immobilized on the inner surface of the cytoplasmic membrane and hydrolyzes bound Guanosine Triphosphate (GTP) to Guanosine Diphosphate (GDP). When the growth factor receptor is activated by the growth factor, it triggers a reactive chain that leads to activation of the exchange activity of guanine nucleotides for Ras. Ras alternates between an activated "on" state with bound GTP (hereinafter "Ras. GTP") and an inactivated "off state with bound GDP. Gtp binds and activates proteins that control cell growth and differentiation.

For example, in the "mitogen-activated protein kinase (MAP kinase) cascade," gtp causes activation of the serine/threonine kinase cascade. Among the several groups of kinases known to require ras gtp to activate themselves is the Raf family. Raf proteins activate "MEKl" and "MEK 2", abbreviations for mitogen-activated ERK-activated kinases (where ERK denotes extracellular signal-regulated protein kinase, another designation for MAPK). MEKl and MEK2 are dual-functional serine/threonine and tyrosine protein kinases, also known as MAP kinase kinases. Gtp activates Raf, thereby activating melk and MEK2, which in turn activates MAP kinase (MAPK). Activation of MAP kinase by mitogens appears to be essential for proliferation, and constitutive activation of this kinase is sufficient to induce cell transformation. Blocking downstream Ras signal transduction, whether induced from cell surface receptors or from oncogenic Ras mutants, can completely inhibit mitogenesis, such as by using dominant negative Raf-1 proteins.

In the control of cell proliferation, the interaction of Raf and Ras is a key regulatory step. To date, no other substrate for MEK has been identified, except for MAPK; however, recent reports indicate that MEK can also be activated by other upstream signaling proteins such as MEK kinase or MEKKl and PKC. Activated MAPK translocates and accumulates in the nucleus where it can phosphorylate and activate transcription factors such as EIk-1 and Sapla, resulting in enhanced expression of genes such as the c-fos gene.

Once activated, Raf and other kinases are at two serine residues adjacent (S for MEK 1)²¹⁸And S²²²) To phosphorylate MEK. These phosphorylations are necessary for the activation of MEK as a kinase. In turn, MEK is separated at two residues separated by a single amino acid: tyrosine Y¹⁸⁵And threonine T¹⁸³Phosphorylate MAP kinase. MEK appears to bind strongly to MAP kinase prior to phosphorylating MAP kinase, suggesting that phosphorylation of MAP kinase by MEK may require a strong interaction between these two proteins in advance. Two factors, the unique specificity of MEK and its requirement for a strong interaction with MAP kinase prior to phosphorylation, suggest that the mechanism of action of MEK may be sufficiently different from that of other protein kinases to allow for selective inhibitors of MEK. It is likely that such inhibitors will act through allosteric mechanisms and not through the more common mechanisms involved in blocking ATP binding sites.

Thus, MEKl and MEK2 are proven and accepted targets for anti-proliferative therapies even when oncogenic mutations do not affect MEK structure or expression. See, for example, U.S. patent publication 2003/0149015 to Barrett et al and 2004/0029898 to Boyle et al.

Several examples of l-substituted-2 (para-substituted-phenylamino) -aryl inhibitors of MEK have been reported. U.S. Pat. Nos. 6,440,966 and 6,750,217 and corresponding publication WO 00/42003 describe carboxylic and hydroxamic esters and N-substituted amide derivatives of sulfonamide-substituted-2 (4-iodophenylamino) -benzoates and N-substituted benzamides that function as MEK inhibitors. The sulfonamides may also be N-substituted.

U.S. patent 6,545,030 and corresponding publication WO 00/42029 describe the MEK inhibitor l-heterocyclyl-2 (4-iodophenylamino) -benzene in which the heterocycle is a 5-membered nitrogen-containing ring such as pyrazole, triazole, oxazole, isoxazole and isoxazolinone (isoxazolinone). More recently, U.S. patent publication 2005/004186 describes related compounds in which the 4-iodo substituent of the' 030 patent is replaced by a very wide variety of groups including alkyl, alkoxy, acyloxy, alkenyl, carbamoyl, carbamoylalkyl, carboxy, carboxyalkyl, N-acylsulfonamido, and others.

U.S. patent 6,469,004 and corresponding publication WO 00/42022 describe a group of carboxylic acid esters and hydroxamic acid esters of heterocyclic fused phenylene compounds (i.e., benzimidazoles, benzoxazoles, benzothiazoles, benzothiadiazoles, quinazolines, etc.). The heterocycle is 7-F-6- (4-iodo-phenylamino) -5-carboxylate, carboxylic acid amide, or hydroxamate. A more recent publication, us 2005/0026970, describes similar compounds in which the 4-iodo substituent is replaced by a very broad class of structures. Related compounds are described in patent publications WO 03/077855, WO 03/77914 and US 2005/0554701. In WO 2005/028426, other examples of 2- (4-iodophenylamino) -phenyl hydroxamates that are reported to be useful as MEK inhibitors can be found.

Patent publication WO 02/06213 and corresponding U.S. application serial No. 10/333,399 (u.s.2004/0054172) describe esters of hydroxy-substituted acids of 1-oxamic acid-2 (4-halophenylamino) -3, 4-difluorobenzene. U.S. patent No. 6,891,066 and corresponding publication WO03/62191 describe similar compounds in which the 4-halogen substituent is replaced by a very broad class of structures. The substituents at the 4-position are methyl, ethyl, ethynyl and 2-hydroxyethyl. Specific related compounds are described in U.S. patent No. 6,770,778.

Patent publication WO 04/083167 (japanese), published on 9/30/2004, discloses more than 2000-but only 400 NMR data-l- (N-substituted sulfonylurea) -2(2, 4-dihalophenylamino) -3, 4-difluorobenzene and asserts that they are useful as MEK inhibitors. Data indicating inhibition of MEK are given for a subset of only 12 compounds. In addition to secondary or tertiary amines, these 12 compounds each contain one of the following groups: n, N-disubstituted sulfonylureas, N-piperazine sulfonamides, N-piperidine sulfonamides or N-pyrrolidine sulfonamides.

The MEK cascade has also been implicated in inflammatory diseases and disorders. U.S. application publication No. 2006/0030610 to Koch et al, U.S. application publication No. 2006/0140872 to Furue et al. This includes both acute and chronic inflammatory conditions. Examples of such conditions are allergic contact dermatitis, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, chronic obstructive pulmonary disorders, psoriasis, multiple sclerosis, asthma, diseases and disorders associated with diabetic complications, and inflammatory complications of the cardiovascular system such as acute coronary syndrome. Inflammatory bowel diseases include Crohn's disease and ulcerative colitis.

MEKl and MEK2 are proven and accepted targets for anti-proliferative therapies even when oncogenic mutations do not affect MEK structure or expression. See, for example, U.S. patent publication 2003/0149015 to Barrett et al and 2004/0029898 to Boyle et al.

Summary of The Invention

Provided herein are compounds of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof:

wherein

Z is H or F;

x is F, Cl, CH₃、CH₂OH、CH₂F、CHF₂Or CF₃；

Y is I, Br, Cl, CF₃、C₁-C₃Alkyl radical, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, cyclopropyl, OMe, OEt, SMe, phenyl or Het wherein Het is a 5 to 10 membered monocyclic or bicyclic heterocyclic group containing 1 to 5 ring heteroatoms independently selected from N, O and S, said heterocyclic group being saturated, olefinic or aromatic; wherein

All said phenyl or Het groups being optionally substituted by F, Cl, Br, I, acetyl, methyl, CN, NO₂、CO₂H、C₁-C₃Alkyl radical, C₁-C₃Alkoxy radical, C₁-C₃alkyl-C (= O) -, C₁-C₃alkyl-C (= S) -, C₁-C₃alkoxy-C (= S) -, C₁-C₃alkyl-C (= O) O-, C₁-C₃alkyl-O- (C = O) -, C₁-C₃alkyl-C (= O) NH-, C₁-C₃alkyl-C (= NH) NH-, C₁-C₃alkyl-NH- (C = O) -, di-C₁-C₃alkyl-N- (C = O) -, C₁-C₃alkyl-C (= O) N (C)₁-C₃Alkyl) -, C₁-C₃alkyl-S (= O)₂NH-or trifluoromethyl;

All of said methyl, ethyl, C₁-C₃Alkyl and cyclopropyl groups optionally substituted with OH;

all of said methyl groups being optionally substituted with 1, 2 or 3F atoms;

R⁰is H, F, Cl, Br, I, CH₃NH-、(CH₃)₂N-、C₁-C₆Alkyl radical, C₁-C₄Alkoxy radical, C₃-C₆Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, phenyl, monosubstituted phenyl, O (C)₁-C₄Alkyl), O-C (= O) (C)₁-C₄Alkyl) or C (= O) O (C)₁-C₄Alkyl groups); wherein

Said alkyl, alkoxy, cycloalkyl, alkenyl, alkynyl and phenyl groups optionally substituted with 1-3 substituents independently selected from F, Cl, Br, I, OH, CN, cyanomethyl, nitro, phenyl and trifluoromethyl;

said C is₁-C₆Alkyl and C₁-C₄Alkoxy radicals optionally being further OCH₃Or OCH₂CH₃Substitution; g is G₁、G₂、R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃(ii) a Wherein

G₁Is optionally substituted by an amino group, C₁-C₃Alkylamino or dialkylamino radical substituted C₁-C₆An alkyl group, said dialkylamino group comprising 2C's which can be the same or different₁-C₄An alkyl group; or

G₁Is C₃-C₈A diaminoalkyl group;

G₂is a saturated, unsaturated or aromatic 5-or 6-membered ring comprising 1-3 ring heteroatoms independently selected from N, O and S, optionally independently selected from F, Cl, OH, O (C)₁-C₃Alkyl), OCH₃、OCH₂CH₃、CH₃C(=O)NH、CH₃C(=O)O、CN、CF₃And 1-3 substituents of a 5-membered aromatic heterocyclic group comprising 1-4 ring heteroatoms independently selected from N, O and S;

R_1aIs methyl, optionally substituted by 1 to 3 fluorine atoms or 1 to 3 chlorine atoms, or OH, cyclopropoxy or C₁-C₃Alkoxy substituted, wherein the ringPropoxy group or said C₁-C₃C of alkoxy radicals₁-C₃Alkyl moiety optionally substituted by a hydroxy or methoxy group, and C₁-C₄All C in alkoxy₃-each alkyl group is optionally further substituted by another OH group;

R_1bis CH (CH)₃)-C_1-3Alkyl or C₃-C₆Cycloalkyl, said alkyl and cycloalkyl groups being optionally independently selected from F, Cl, Br, I, OH, OCH₃And 1-3 substituents of CN;

R_1cis (CH)₂)_nO_mR'; wherein

m is 0 or 1; and wherein

When m is 0, n is 1 or 2;

when m is 1, n is 2 or 3;

r' is C₁-C₆Alkyl, optionally independently selected from F, Cl, OH, OCH₃、OCH₂CH₃And C₃-C₆1-3 substituents of cycloalkyl;

R_1dis C (A), (A') (B) -; wherein

B is H or C_1-4Alkyl, optionally substituted with one or two OH groups;

a and A' are independently H or C_1-4Alkyl, optionally substituted with one or two OH groups; or

A and A' together with the carbon atom to which they are attached form a 3-6 membered saturated ring;

R_1eis that

Wherein

q is 1 or 2;

R₂and R₃Each independently is H, F, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl or methylsulfonyl;

R₄Is H, F, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, N-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl, methylsulfonyl, nitro, acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazole, 1,3, 4-thiadiazole, 5-methyl-l, 3, 4-thiadiazole, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl and N-pyrrolidinylcarbonylamino;

R₅is H, F, Cl or methyl;

R₆is H, F, Cl or methyl;

Ar₁is that

Wherein

U and V are independently N, CR₂Or CR₃；

R₂、R₃And R₄Independently H, F, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl, acetamidoA group selected from the group consisting of an amidino group, a cyano group, a carbamoyl group, a methylcarbamoyl group, a dimethylcarbamoyl group, a l,3, 4-oxadiazol-2-yl group, a 5-methyl-l, 3, 4-oxadiazolyl group, a 1,3, 4-thiadiazolyl group, a 5-methyl-l, 3, 4-thiadiazolyl group, an lH-tetrazolyl group, an N-morpholinylcarbonylamino group, an N-morpholinylsulfonyl group, an N-pyrrolidinylcarbonylamino group and a methanesulfonyl group;

R₅and R₆Independently H, F, Cl or methyl;

Ar₂is that

Wherein

The dotted line represents an alternative formal position of the second ring double bond;

U is-S-, -O-or-N =, and wherein

When U is-O-or-S-, V is-CH =, -CCl = or-N =;

when U is-N = V is-CH =, -CCl =, or-N =;

R₇is H or methyl;

R₈is H, acetamido, methyl, F or Cl;

Ar₃is that

Wherein

U is-NH-, -NCH₃-or-O-;

R₇and R₈Independently H, F, Cl or methyl.

In some embodiments, the present invention provides a compound of formula I selected from the following compounds:

in some embodiments, the present invention provides a compound of formula I selected from:

wherein the 2-OH carbon is in the R configuration.

wherein the 2-OH carbon is in the S configuration.

In some embodiments, the present invention provides compositions comprising a compound of formula I selected from the compounds shown below, wherein the 2-OH carbon is in the R configuration, said compositions being substantially free of the S-isomer.

In some embodiments, the present invention provides compositions comprising a compound of formula I selected from the compounds shown below, wherein the 2-OH carbon is in the S configuration, said compositions being substantially free of the R-isomer.

In some embodiments, the present invention provides a compound of formula I, wherein Y is phenyl, pyridinyl, or pyrazolyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein Y is substituted phenyl, pyridyl, or A pyrazolyl group. In another subgeneric embodiment, the invention provides a compound of formula I wherein Y is Br or I. In a subgeneric embodiment, the invention provides a compound of formula I, wherein G is 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, or 4-piperidinyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is 1-piperazinyl or 2-piperazinyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is morpholinyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-2-aminoethyl. In a subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-3-amino-N-propyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1, 2 or 3. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is (CH)₃CH₂)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1 or 2. In a more specific subgeneric embodiment, the invention provides a compound of formula I, wherein G is 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, or 4-piperidinyl; r^oIs H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is 1-piperazinyl or 2-piperazinyl; r ^oIs H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is morpholinyl; r^oIs H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-2-aminoethyl; r^oIs H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-3-amino-N-propyl; r^oIs H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I, whereinG is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1, 2 or 3; r^oIs H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is (CH)₃CH₂)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1 or 2; r^oIs H, halogen or methoxy; x is F; and Y is I.

In some embodiments, the present invention provides pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.

In some embodiments, the present invention provides a composition comprising a compound selected from:or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier. In some embodiments, the compound is in the R configuration. In some embodiments, the compound is in the R configuration, substantially free of the S-isomer. In some embodiments, the compound is in the S configuration.

In some embodiments, the compound is in the S configuration, substantially free of the R-isomer. In some embodiments, the compound is:in some embodiments, the compound is:

the present invention also relates to N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (also referred to herein as "compound a" and "N- (-) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide"):exhibit a specific powder X-ray diffraction pattern. In some embodiments, the powder X-ray diffraction pattern comprises at least 50% of the peaks shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 70% of the peaks shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 90% of the peaks shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5. Compound a has been characterized as the "S" isomer by making R and S-MTPA esters on secondary alcohols and comparing proton chemical shift differences. See, e.g., Dale, j.a.; mosher, h.s., j.am.chem.soc.,1973,95,512 and Ohtani et al, j.am.chem.soc, 1991,113,4092.

The invention also relates to N- (R) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (also referred to herein as "compound B" and "N- (+) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide"):compound B has been characterized as the "R" isomer by making R and S-MTPA esters on secondary alcohols and comparing proton chemical shift differences. See, e.g., Dale, j.a.; mosher, h.s., j.am.chem.soc.,1973,95,512 and Ohtani et al, j.am.chem.soc, 1991,113,4092.

The invention also relates to N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -lCrystalline polymorph a of- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide:it shows a specific differential scanning calorimetry pattern. In some embodiments, the differential scanning calorimetry map is substantially the same as the differential scanning calorimetry map shown in fig. 6.

The present invention also relates to pharmaceutical compositions comprising an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and a pharmaceutically acceptable carrier or vehicle.

In some embodiments, crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is used to treat or prevent cancer or an inflammatory disease. The present invention further relates to a method of treating or preventing cancer or inflammatory diseases comprising administering to a subject in need thereof an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

In other aspects, the invention relates to pharmaceutical compositions comprising an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. Such compositions may contain adjuvants, excipients and preservatives, agents for delaying absorption, fillers, binders, adsorbents, buffers, disintegrants, solubilizers, other carriers, and other inert ingredients. Methods of formulating such compositions are well known in the art.

In other aspects, the invention relates to pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In some embodiments, the pharmaceutical composition is in a form suitable for oral administration. In further or other embodiments, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release formulation, solution, suspension, sterile solution, suspension or emulsion for parenteral injection, ointment or cream for topical administration, or suppository for rectal administration. In further or other embodiments, the pharmaceutical composition is in unit dosage form suitable for single administration of an accurate dose. In further or other embodiments the amount of the compound of formula I is in the range of about 0.001 to about 1000mg/kg body weight/day. In further or other embodiments the amount of the compound of formula I is in the range of about 0.5 to about 50 mg/kg/day. In further or other embodiments the amount of the compound of formula I is from about 0.001 to about 7 g/day. In further or other embodiments the amount of the compound of formula I is from about 0.002 to about 6 g/day. In further or other embodiments the amount of the compound of formula I is from about 0.005 to about 5 g/day. In further or other embodiments the amount of the compound of formula I is from about 0.01 to about 5 g/day. In further or other embodiments the amount of the compound of formula I is from about 0.02 to about 5 g/day. In further or other embodiments the amount of the compound of formula I is from about 0.05 to about 2.5 g/day. In further or other embodiments the amount of the compound of formula I is from about 0.1 to about 1 g/day.

In further or alternative embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate. In further or alternative embodiments, dosage levels above the upper limit of the range recited above may be necessary. In further or other embodiments, the compound of formula I is administered once daily in a single dose. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily.

In some embodiments, the pharmaceutical composition is for administration to a mammal. In further or other embodiments, the mammal is a human.

In further or other embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient and/or adjuvant. In further or other embodiments, the pharmaceutical composition further comprises at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins; antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormone/anti-hormone therapeutics, and hematopoietic growth factors. In further or other embodiments, the therapeutic agent is paclitaxel, bortezomib, or both. In further or other embodiments, the pharmaceutical composition is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable salt of the compound of formula I.

Provided herein comprise compounds selected fromAnd the use of a composition comprising a compound selected fromA method of making a composition of the compound of (1). In some embodiments, the 2-OH carbon on the compound is in the R configuration. In some embodiments, the 2-OH carbon on the compound is in the S configuration. In some embodiments, the composition is substantially free of the S-isomer of the compound. In some embodiments, the composition is substantially free of the R-isomer of the compound. In some embodiments, the compound comprises less than 10% of the compoundThe S-isomer of the compound. In some embodiments, the compound comprises less than 10% of the R-isomer of the compound. In some embodiments, the compound comprises less than 5% of the S-isomer of the compound. In some embodiments, the compound comprises less than 5% of the R-isomer of the compound. In some embodiments, the compound comprises less than 1% of the S-isomer of the compound. In some embodiments, the compound comprises less than 1% of the R-isomer of the compound.

Also provided herein are pharmaceutical compositions comprising from about 1 mg to about 100mg of a compound having the structure: And with a composition comprising about 1-100mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition allows for modified release of the compound. In some embodiments, the composition allows for sustained release of the compound. In some embodiments, the composition allows for delayed release of the compound. In some embodiments, the compound is present in an amount of about 1-50 mg. In some embodiments, the compound is present in an amount of about 1-10 mg. In some embodiments, the compound is present in an amount of about 10-20 mg. In some embodiments, the compound is present in an amount of about 20-40 mg. In some embodiments, the compound is present in an amount of about 40-50 mg.

Also provided herein are pharmaceutical compositions comprising from about 1 to 50mg of a compound having the structure:and with a composition comprising about 1-50mg of a compound having the structure:for the treatment of cancer or inflammationThe method of (4), wherein said composition allows for modified release of said drug. In some embodiments, the composition further comprises microcrystalline cellulose. In some embodiments, the composition further comprises croscarmellose sodium. In some embodiments, the composition further comprises sodium lauryl sulfate. In some embodiments, the composition further comprises magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 1mg of a compound having the structure:the composition of (1). In some embodiments, the composition further comprises about 222.2mg microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 10mg of a compound having the structure:and with a composition comprising about 10mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 213.2mg microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 20mg of a compound having the structure:and with a composition comprising about 20mg ofA compound of the structure: The composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 203.2mg microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 40mg of a compound having the structure:and with a composition comprising about 40mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 183.2mg of microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 0.4 wt% of a compound having the structure:and about 99.6% by weight of a pharmaceutically acceptable carrier or vehicle and a pharmaceutically acceptable excipient or vehicle comprising about 0.4% by weight of a compound having the structure: And about 99.6% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In some embodiments, the microcrystalline cellulose comprises about 92.6% by weight of the composition. In some embodiments, the composition further comprises about 5% by weight croscarmellose sodium. In some embodiments, the composition further comprises about 1% by weight sodium lauryl sulfate. In some embodiments, the composition further comprises about 1% by weight magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 4.2 wt% of a compound having the structure:and about 95.8% by weight of a pharmaceutically acceptable carrier or vehicle and a pharmaceutically acceptable excipient or vehicle comprising about 4.2% by weight of a compound having the structure:and about 95.8% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In some embodiments, the microcrystalline cellulose comprises about 88.8% by weight of the composition. In some embodiments, the composition further comprises about 5% by weight croscarmellose sodium. In some embodiments, the composition further comprises about 1% by weight sodium lauryl sulfate. In some embodiments, the composition further comprises about 1% by weight magnesium stearate.

Also provided herein are compositions comprising from about 2% to about 10% by weight of a compound having the structure:and about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle and a pharmaceutically acceptable excipient or vehicle comprising about 2% to about 10% by weight of a compound having the structure:and about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle for treating cancer or inflammation. In some embodiments, the pharmaceutically acceptable carrier or vehicle further comprises microcrystalline cellulose. In some embodiments, the microcrystalline cellulose comprises from about 85% to about 95% by weight of the composition. In some embodiments, the composition further comprises from about 1% to about 6% by weight croscarmellose sodium. In some embodiments, the composition further comprises from about 0.1% to about 2% by weight sodium lauryl sulfate. In some embodiments, the composition further comprises from about 0.25% to about 1.5% by weight magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 1mg of a compound having the structure:and with a composition comprising about 1mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 222.2mg microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 10mg of a compound having the structure:and with a composition comprising about 10mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 213.2mg microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg of lauryl sulfuric acidSodium. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 20mg of a compound having the structure:and with a composition comprising about 20mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 203.2mg microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 40mg of a compound having the structure: And with a composition comprising about 40mg of a compound having the structure:the composition of (a) is used in a method of treating cancer or inflammation. In some embodiments, the composition further comprises about 183.2mg of microcrystalline cellulose. In some embodiments, the composition further comprises about 12.0mg croscarmellose sodium. In some embodiments, the composition further comprises about 2.4mg sodium lauryl sulfate. In some embodiments, the composition further comprises about 2.4mg magnesium stearate.

Also provided herein is a pharmaceutical composition comprising about 0.4 wt% of a compound having the structure:and about 99.6% by weight of a pharmaceutically acceptable carrier or vehicle and pharmaceutical compositions containing the sameAbout 0.4 wt% of a compound having the structureAnd about 99.6% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In some embodiments, the microcrystalline cellulose comprises about 92.6% by weight of the composition. In some embodiments, the composition further comprises about 5% by weight croscarmellose sodium. In some embodiments, the composition further comprises about 1% by weight sodium lauryl sulfate. In some embodiments, the composition further comprises about 1% by weight magnesium stearate.

Also provided herein are compositions comprising from about 2% to about 10% by weight of a compound having the structure:and about 98% to about 90% by weight of a pharmaceutically acceptable carrierA composition for receiving a carrier or vehicle and a pharmaceutical composition comprising from about 2% to about 10% by weight of a compound having the structure:and about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In some embodiments, the microcrystalline cellulose comprises from about 85% to about 95% by weight of the composition. In some embodiments, the composition further comprises from about 1% to about 6% by weight of croscarmellose sodium. In some embodiments, the composition further comprises from about 0.1% to about 2% by weight sodium lauryl sulfate. In some embodiments, the composition further comprises from about 0.25% to about 1.5% by weight magnesium stearate.

Also provided herein is a crystalline polymorph a of N- (-) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide that exhibits a powder X-ray diffraction pattern comprising at least 50% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5, and compositions comprising this compound. In some embodiments, the crystalline polymorph a, wherein the powder X-ray diffraction pattern comprises at least 70% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 90% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5. In some embodiments, the crystalline polymorph has a melting point onset (melting point onset) of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph is substantially free of solvent.

Also provided herein is a crystalline polymorph a of N- (-) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide that exhibits a differential scanning calorimetry trace substantially the same as the differential scanning calorimetry trace shown in figure 6, and compositions comprising this compound. In some embodiments, the crystalline polymorph has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph of claim 67 or 68, wherein said crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph of any one of claims 67-69, wherein the crystalline polymorph is substantially free of solvent.

Also provided herein are polymorphs of N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide prepared by a process comprising the step of crystallizing amorphous N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, and compositions comprising this compound. In some embodiments, the crystallizing step comprises crystallizing from a mixture of ethyl acetate and heptane. In some embodiments, the ratio of the mixture of ethyl acetate and heptane is from about 1-4 parts ethyl acetate to about 2-10 parts heptane. In some embodiments, the ratio of the mixture of ethyl acetate and heptane is about 2 parts ethyl acetate to about 5 parts heptane.

Also provided herein are methods of inhibiting a MEK enzyme comprising contacting the MEK enzyme with a compound or composition described herein, wherein the compound is present in an amount sufficient to inhibit the enzyme by at least 25%. In some embodiments, the MEK enzyme is a MEK kinase. In some embodiments, the contacting occurs within a cell.

Also provided herein are methods of treating a MEK-mediated disorder in a subject suffering from such disorder comprising administering to the subject an effective amount of a compound or composition described herein. In some embodiments, the MEK inhibitor is administered in combination with other therapies. In some embodiments, the other therapy is radiation therapy, non-MEK kinase inhibitor therapy, chemotherapy, surgery, glucocorticoids, methotrexate, biological response modifiers, or any combination thereof. In some embodiments, the MEK mediated disorder is selected from an inflammatory disease, an infection, an autoimmune disease, stroke, ischemia, a cardiac disorder, a neurological disorder, a fibrotic disorder, a proliferative disorder, a hyperproliferative disorder, a tumor, a leukemia, a neoplasm (neoplasms), a cancer, a carcinoma, a metabolic disease, and a malignant disease. In some embodiments, the MEK mediated disorder is a hyperproliferative disease. In some embodiments, the MEK mediated disorder is a cancer, tumor, leukemia, neoplasm, or carcinoma. In some embodiments, the MEK mediated disorder is an inflammatory disease. In some embodiments, the inflammatory disease is rheumatoid arthritis or multiple sclerosis.

Also provided herein are methods of treating or preventing a proliferative disease in a subject, comprising administering to the subject an effective amount of a compound or composition described herein. In some embodiments, the proliferative disease is cancer, psoriasis, restenosis, disease, or atherosclerosis. In some embodiments, the proliferative disease is cancer. In some embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, colorectal cancer, leukemia, myeloid leukemia, glioblastoma, follicular lymphoma (folliculular lymphoma), pre-B acute leukemia, chronic lymphocytic B leukemia, gastric cancer, mesothelioma, or small cell lung cancer. In some embodiments, the method further comprises administering at least one therapeutic agent. In some embodiments, this step comprises administering at least one additional cancer therapy. In some embodiments, the other therapy is radiation therapy, non-MEK kinase inhibitor therapy, chemotherapy, surgery, glucocorticoids, methotrexate, biological response modifiers, or any combination thereof.

Also provided herein are methods of treating or preventing an inflammatory disease in a subject comprising administering to the subject an effective amount of a composition comprising a compound described herein. In some embodiments, the inflammatory disease is rheumatoid arthritis or multiple sclerosis.

Also provided herein are methods of degenerating, inhibiting the growth of, or killing a cancer cell comprising contacting the cell with a compound or composition described herein in an amount effective to degenerate, inhibit the growth of, or kill a cancer cell. In some embodiments, the cancer cell comprises a brain cancer cell, a breast cancer cell, a lung cancer cell, an ovarian cancer cell, a pancreatic cancer cell, a prostate cancer cell, a renal cancer cell, a gastric cancer cell, or a colorectal cancer cell.

Also provided herein are methods of inhibiting tumor size increase, reducing tumor size, reducing tumor proliferation, or preventing tumor proliferation in a subject, comprising administering to the subject an effective amount of a compound or composition described herein to inhibit tumor size increase, reduce tumor size, reduce tumor proliferation, or prevent tumor proliferation. In some embodiments, the tumor is present in the brain, breast, lung, ovary, pancreas, prostate, kidney, stomach, colon, or rectum.

Also provided herein are methods of treating or preventing ankylosing spondylitis, gout, tendonitis, bursitis, or sciatica, comprising administering to a subject in need thereof an effective amount of a compound of formula (I):

Wherein:

z is H or F;

x is F, Cl, CH₃、CH₂OH、CH₂F、CHF₂Or CF₃；

Y is I, Br, Cl, CF₃、C₁-C₃Alkyl radical, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, cyclopropyl, OMe, OEt, SMe, phenyl or Het wherein Het is 1-5 comprising independently selected from N, O and SA 5 to 10 membered monocyclic heterocyclic group or bicyclic heterocyclic group of a ring heteroatom, said heterocyclic groups being saturated, olefinic or aromatic; wherein

all of said methyl groups being optionally substituted with 1, 2 or 3F atoms;

Said C is₁-C₆Alkyl and C₁-C₄Alkoxy radicals optionally being further OCH₃Or OCH₂CH₃Substitution;

g is G₁、G₂、R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃(ii) a Wherein

G₁Is C₃-C₈A diaminoalkyl group;

R_1ais methyl, optionally substituted by 1 to 3 fluorine atoms or 1 to 3 chlorine atoms, or OH, cyclopropoxy or C₁-C₃Alkoxy substituted, wherein said cyclopropoxy group or said C₁-C₃C of alkoxy radicals₁-C₃Alkyl moiety optionally substituted by a hydroxy or methoxy group, and C₁-C₄All C in alkoxy₃-each alkyl group is optionally further substituted by another OH group;

R_1bis CH (CH)₃)-C_1-3Alkyl or C₃-C₆Cycloalkyl, said alkyl and cycloalkyl groups being optionally independently selected from F, Cl, Br, I、OH、OCH₃And 1-3 substituents of CN;

R_1cIs (CH)₂)_nO_mR'; wherein

m is 0 or 1; and wherein

When m is 0, n is 1 or 2;

when m is 1, n is 2 or 3;

R_1dis C (A), (A') (B) -; wherein

B is H or C_1-4Alkyl, optionally substituted with one or two OH groups;

R_1eis that

Wherein

q is 1 or 2;

R₄is H, F, Cl、Br、CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, N-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl, methylsulfonyl, nitro, acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazole, 1,3, 4-thiadiazole, 5-methyl-l, 3, 4-thiadiazole, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl and N-pyrrolidinylcarbonylamino;

R₅Is H, F, Cl or methyl;

R₆is H, F, Cl or methyl;

Ar₁is that

Wherein

U and V are independently N, CR₂Or CR₃；

R₂、R₃And R₄Independently H, F, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, N-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl, acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazolyl, 1,3, 4-thiadiazolyl, 5-methyl-l, 3, 4-thiadiazolyl, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrolidinylcarbonylamino and methanesulfonyl;

R₅and R₆Independently H, F, Cl or methyl;

Ar₂is that

Wherein

u is-S-, -O-or-N =, and wherein

When U is-O-or-S-, V is-CH =, -CCl = or-N =;

when U is-N = V is-CH =, -CCl =, or-N =;

R₇is H or methyl;

R₈is H, acetamido, methyl, F or Cl;

Ar₃is that

Wherein

U is-NH-, -NCH₃-or-O-;

R₇and R₈Independently H, F, Cl or methyl.

In some embodiments, the compound is selected from:

in some embodiments, the compound is selected fromWherein the 2-OH carbon is in the R configuration. In some cases In embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected fromWherein the 2-OH carbon is in the S configuration. In some embodiments, the compound isIn some embodiments, the compound is

Also provided herein are methods of treating gastric cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating leukemia melanoma (leukaemiam melanoma) or hepatoma by administering a therapeutically effective amount of a compound or composition described herein.

Also provided herein are methods of treating non-small cell lung cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating colon cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating CNS cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating ovarian cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating renal cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating prostate cancer by administering a therapeutically effective amount of a compound or composition described herein. Also provided herein are methods of treating breast cancer by administering a therapeutically effective amount of a compound or composition described herein. In various embodiments, the methods further comprise administering at least one additional therapeutic agent. In some embodiments, at least one additional cancer therapy is administered. In some embodiments, the other cancer therapy is radiation therapy, chemotherapy, surgery, or any combination thereof.

Also provided herein are methods of treating or preventing psoriasis by administering a therapeutically effective amount of a compound or composition described herein in a topical dosage form.

In various embodiments, the composition is administered orally. In some embodiments, the composition is administered once daily or twice daily. In some embodiments, the composition is administered once daily for at least one week.

In some embodiments, when the composition is administered orally, the T of the compound is achieved between 1 hour and 3 hours after administration of the composition to a fasted subject_max. In some embodiments, the compound reaches C on day 1 when administered to a subject_maxAbout 0.01 μ g/ml to about 1.0 μ g/ml. In some embodiments, the compound reaches C on day 1 when administered to a subject_maxFrom about 0.01. mu.g/ml to about 0.8. mu.g/ml. In some embodiments, the compound reaches C on day 1 when administered to a subject_maxFrom about 0.03. mu.g/ml to about 0.5. mu.g/ml. In some embodiments, the AUC of the compound over a 0-12 hour period is about 0.1 μ g hr/mL to about 5.0 μ g hr/mL. In some embodiments, the AUC of the compound is from about 0.1 μ g hr/mL to about 4.0 μ g hr/mL. In some embodiments, the AUC of the compound is about 0.5 μ g hr/mL to about 3.0 μ g hr/mL. In some embodiments, T of the compound _maxBetween 0.5 and 5.0 hours. In some embodiments, T of the compound_maxBetween 1.0 and 3.0 hours. In some embodiments, T of the compound_maxBetween 1.0 and 2.5 hours. In some embodiments, the plasma concentration of the compound is greater than about 0.01mg/mL 5 hours after a single administration. In some embodiments, the plasma concentration of the compound is greater than about 0.01mg/mL after 10 hours of a single administration. In some embodiments, the plasma concentration of the compound is greater than about 0.01mg/mL 15 hours after a single administration.

In some embodiments, when administered to a group of 10 subjects, the compound reaches a mean C on day 1_maxAbout 0.01 μ g/ml to about 1.0 μ g/ml. In some embodiments, when administered to a group of 10 subjects, the compound reaches a mean C on day 1_maxFrom about 0.01. mu.g/ml to about 0.8. mu.g/ml. In some embodiments, when administered to a group of 10 subjects, the compound reaches a mean C on day 1_maxFrom about 0.03. mu.g/ml to about 0.5. mu.g/ml. In some embodiments, the average AUC of the compound is about 0.1 μ g hr/mL to about 5.0 μ g hr/mL. In some embodiments, the average AUC of the compound is about 0.1 μ g hr/mL to about 4.0 μ g hr/mL. In some embodiments, the average AUC of the compound is about 0.5 μ g hr/mL to about 3.0 μ g hr/mL. In some embodiments, the average T of the compound _maxBetween 0.5 and 5.0 hours. In some embodiments, the average T of the compound_maxBetween 1.0 and 3.0 hours. In some embodiments, the average T of the compound_maxBetween 1.0 and 2.5 hours.

Also provided herein are methods of reducing tumor volume by administering the compounds and compositions described herein. In some embodiments, the tumor volume is reduced by at least about 25% after 5 days of daily administration of the drug. In some embodiments, the tumor volume is reduced by at least about 50% after 5 days of daily administration of the drug. In some embodiments, the tumor volume is reduced by at least about 20-70% after 5 days of daily administration of the drug. In some embodiments, the tumor volume is reduced by at least about 25% after 15 days of daily administration of the drug. In some embodiments, the tumor volume is reduced by at least about 50% after 15 days of daily administration of the drug. In some embodiments, the tumor volume is reduced by at least about 20-70% after 15 days of daily administration of the drug. In some embodiments, the tumor volume is reduced by at least about 25% after 30 days of daily administration of the medicament. In some embodiments, the tumor volume is reduced by at least about 50% after 30 days of daily administration of the medicament. In some embodiments, the tumor volume is reduced by at least about 20-70% after 30 days of daily administration of the medicament.

Also provided herein are methods of inhibiting tumor growth by administering the compounds and compositions described herein. In some embodiments, the tumor growth is inhibited by at least about 20% following administration of the agent. In some embodiments, the tumor growth is inhibited by at least about 40% following administration of the agent. In some embodiments, the tumor growth is inhibited by at least about 60% following administration of the agent. In some embodiments, the tumor growth is inhibited by at least about 80% following administration of the agent. In some embodiments, the tumor growth is inhibited by about 20% to about 100% following administration of the medicament. In some embodiments, the tumor growth is substantially inhibited following administration of the agent.

In some embodiments, the composition is administered twice daily. In some embodiments, the composition is administered once daily.

In some embodiments, the MEK inhibitor does not interfere with the co-administration of another tumor inhibitor.

In some embodiments, the composition is in the form of a tablet, capsule, soft capsule (gel cap), lozenge, pellet, or granule. In some embodiments, the composition is in the form of a capsule or tablet dosage form having a total weight of about 50mg to about 1000 mg. In some embodiments, the composition is in the form of a capsule or tablet having a total weight selected from 50mg, 75mg, 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, and 500 mg. In some embodiments, the composition is in the form of a capsule or tablet having a total weight of about 240 mg.

In some embodiments, the composition further comprises at least one filler selected from the group consisting of microcrystalline cellulose, silicified microcrystalline cellulose, lactose, compressible sugar, xylitol, sorbitol, mannitol, pregelatinized starch, maltodextrin, calcium phosphate, calcium carbonate, starch, and calcium silicate.

In some embodiments, the composition further comprises at least one disintegrant selected from the group consisting of croscarmellose sodium, sodium starch glycolate, crospovidone, methylcellulose, alginic acid, sodium alginate, starch derivatives, bentonite (beta), and magnesium aluminum silicate.

In some embodiments, the composition further comprises at least one lubricant selected from the group consisting of magnesium stearate, metallic stearates, talc, sodium stearyl fumarate, and stearic acid.

In some embodiments, the composition further comprises at least one wetting agent or surfactant selected from sodium lauryl sulfate, glycerin, sorbitan oleate, sorbitan stearate, polyoxyethylated sorbitan laurate, palmitate, stearate, oleate or hexaoleate (hexaoleate), polyoxyethylene stearyl alcohol, and sorbitan monolaurate.

Provided herein are compositions in the form of capsules or tablets that release at least 60% of the drug in 30 minutes using United States Pharmacopeia (USP) apparatus II at 50rpm with 1% sodium lauryl sulfate in water as the dissolution medium. In some embodiments, the composition is in the form of a capsule or tablet, and the capsule or tablet releases about 60-100% of the drug within 30 minutes using United States Pharmacopeia (USP) apparatus II at 50rpm with 1% sodium lauryl sulfate in water as the dissolution medium. In some embodiments, the composition is in the form of a capsule or tablet, and the capsule or tablet releases about 60-90% of the drug within 30 minutes using United States Pharmacopeia (USP) apparatus II at 50rpm with 1% sodium lauryl sulfate in water as the dissolution medium. In some embodiments, the composition is in the form of a capsule or tablet, and the capsule or tablet releases about 60-80% of the drug within 30 minutes using United States Pharmacopeia (USP) apparatus II at 50rpm with 1% sodium lauryl sulfate in water as the dissolution medium.

Also provided herein are batches of capsules or tablets, each comprising from about 1 to about 50mg of a compound described herein, and having a content uniformity USP acceptance value (acceptance value) of less than about 15.

Method of treatment

The present invention relates to a method of treating or preventing cancer comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a compound of formula (I) as described herein. In various embodiments, compounds and compositions useful in these methods are as described in the genus of formula (I) or as described in any subgenus or species by way of example within the scope of formula (I) throughout this application.

The present invention relates to a method of treating or preventing an inflammatory disease comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a compound of formula (I) as described herein. In various embodiments, compounds and compositions useful in these methods are as described in the genus of formula (I) or as described in any subgenus or species by way of example within the scope of formula (I) throughout this application.

In some embodiments, the present invention relates to a method of treating or preventing ankylosing spondylitis, gout, tendonitis, bursitis, or sciatica, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a compound of formula (I) as described herein. In various embodiments, compounds and compositions useful in these methods are of the genus of formula (I) or as described by any subgenus or species by way of example within the scope of formula (I) throughout this application.

In some aspects, the invention also relates to a method of treating a disease in a subject suffering from said disease, comprising administering to said subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof.

In other aspects, the invention relates to methods of treating a disorder in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof.

In other aspects, the invention relates to methods of treating conditions in a human comprising administering to the mammal a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof.

In other aspects, the invention relates to methods of treating hyperproliferative disorders in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof.

In other aspects, the invention relates to a method of treating an inflammatory disease, condition, or disorder in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.

In other aspects, the invention relates to methods of treating a disorder or condition modulated by the MEK cascade in a mammal, including a human, comprising administering to said mammal an amount of a compound of formula I, or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof, effective to modulate said cascade. The appropriate dosage for a particular patient can be determined by one skilled in the art according to known methods.

Inhibition of MEK enzymes

In other aspects, the invention relates to methods of inhibiting MEK enzymes. In some embodiments, the methods comprise contacting the MEK enzyme with an amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof, sufficient to inhibit the enzyme, wherein the enzyme is inhibited. In further or other embodiments, the enzyme is inhibited by at least about 1%. In further or other embodiments, the enzyme is inhibited by at least about 2%. In further or other embodiments, the enzyme is inhibited by at least about 3%. In further or other embodiments, the enzyme is inhibited by at least about 4%. In further or other embodiments, the enzyme is inhibited by at least about 5%. In further or other embodiments, the enzyme is inhibited by at least about 10%. In further or other embodiments, the enzyme is inhibited by at least about 20%. In further or other embodiments, the enzyme is inhibited by at least about 25%. In further or other embodiments, the enzyme is inhibited by at least about 30%. In further or other embodiments, the enzyme is inhibited by at least about 40%. In further or other embodiments, the enzyme is inhibited by at least about 50%. In further or other embodiments, the enzyme is inhibited by at least about 60%. In further or other embodiments, the enzyme is inhibited by at least about 70%. In further or other embodiments, the enzyme is inhibited by at least about 75%. In further or other embodiments, the enzyme is inhibited by at least about 80%. In further or other embodiments, the enzyme is inhibited by at least about 90%. In further or other embodiments, the enzyme is substantially completely inhibited. In further or other embodiments, the MEK enzyme is a MEK kinase. In further or other embodiments, the MEK enzyme is MEK 1. In further or other embodiments, the MEK enzyme is MEK 2. In further or other embodiments, the contacting occurs intracellularly. In further or other embodiments, the cell is a mammalian cell. In further or other embodiments, the mammalian cell is a human cell. In further or other embodiments, the MEK enzyme is inhibited with a composition comprising a pharmaceutically acceptable salt of a compound of formula I.

MEK mediated disorders

In other aspects, the invention relates to methods of treating a MEK-mediated disorder in a subject suffering from such disorder comprising administering to the subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof. In some embodiments, the composition comprising a compound of formula I is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically, or rectally. In some embodiments, the pharmaceutical composition is in a form suitable for oral administration. In further or other embodiments, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release formulation, solution, suspension, sterile solution, suspension or emulsion for parenteral injection, ointment or cream for topical administration, or suppository for rectal administration. In further or other embodiments, the pharmaceutical composition is in unit dosage form suitable for single administration of an accurate dose. In further or other embodiments, the pharmaceutical composition further comprises a pharmaceutical carrier, excipient and/or adjuvant.

In further or other embodiments, the amount of the compound of formula I ranges from about 0.001 to about 1000mg/kg body weight/day. In further or other embodiments, the amount of the compound of formula I is in the range of about 0.5 to about 50 mg/kg/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.001 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.01 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.02 to about 5 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.05 to about 2.5 g/day. In further or other embodiments, the amount of the compound of formula I is from about 0.1 to about 1 g/day. In further or alternative embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate. In further or alternative embodiments, dosage levels above the upper limit of the range recited above may be necessary.

In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the subject having a MEK mediated disorder is a mammal. In further or other embodiments, the subject is a human.

In some embodiments, the composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins; antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormone/anti-hormone therapeutics, and hematopoietic growth factors. In further or other embodiments, the therapeutic agent is selected from paclitaxel, bortezomib, or both.

In some embodiments, the MEK mediated disorder is selected from inflammatory diseases, infections, autoimmune diseases, stroke, ischemia, cardiac disorders, neurological disorders, fibrotic disorders, proliferative disorders, hyperproliferative disorders, non-cancer hyperproliferative disorders, tumors, leukemias, tumors, cancers, metabolic diseases, malignant diseases, vascular restenosis, psoriasis, atherosclerosis, rheumatoid arthritis, osteoarthritis, heart failure, chronic pain, neuropathic pain, dry eye, angle-closure glaucoma and open angle glaucoma. In further or other embodiments, the MEK mediated disorder is an inflammatory disease. In further or other embodiments, the MEK mediated disorder is a hyperproliferative disease. In further or other embodiments, the MEK mediated disorder is selected from a tumor, a leukemia, a neoplasm, a cancer, a carcinoma and a malignant disease. In further or other embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, stomach cancer, kidney cancer, colorectal cancer, or leukemia. In further or other embodiments, the fibrotic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid formation, interstitial nephritis, or pulmonary fibrosis. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

Obtain the effect

In other aspects, the invention relates to methods of achieving an effect in a patient comprising administering to the patient an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof, wherein the effect is selected from the group consisting of inhibition of various cancers, immune diseases and inflammatory diseases. In some embodiments, the effect is inhibition of various cancers. In further or other embodiments, the effect is suppression of an immune disease. In further or other embodiments, the effect is inhibition of an inflammatory disease.

In some embodiments, the composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent.

In some embodiments, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly, or by infusion), topically, or rectally. In further or other embodiments, the amount of the compound of formula I ranges from about 0.001 to about 1000mg/kg body weight/day. In further or other embodiments, the amount of the compound of formula I is in the range of about 0.5 to about 50 mg/kg/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.001 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.01 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.02 to about 5 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.05 to about 2.5 g/day. In further or other embodiments, the amount of the compound of formula I is from about 0.1 to about 1 g/day. In further or alternative embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate. In further or alternative embodiments, dosage levels above the upper limit of the range recited above may be necessary.

In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the individual having cancer is a mammal. In further or other embodiments, the subject is a human. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

In other aspects, the invention relates to a method of degenerating, inhibiting the growth of, or killing a cancer cell comprising contacting the cell with a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof, in an amount effective to degenerate, inhibit the growth of, or kill the cell. In some embodiments, the cancer cell comprises a brain cancer cell, a breast cancer cell, a lung cancer cell, an ovarian cancer cell, a pancreatic cancer cell, a prostate cancer cell, a renal cancer cell, or a colorectal cancer cell. In further or other embodiments, the composition is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is paclitaxel, bortezomib, or both. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins; antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormone/anti-hormone therapeutics, and hematopoietic growth factors. In some embodiments, the cancer cell is degenerated. In further or other embodiments, 1% of the cancer cells are degenerated. In further or other embodiments, 2% of the cancer cells are degenerated. In further or other embodiments, 3% of the cancer cells are degenerated. In further or other embodiments, 4% of the cancer cells are degenerated. In further or other embodiments, 5% of the cancer cells are degenerated. In further or other embodiments, 10% of the cancer cells are degenerated. In further or other embodiments, 20% of the cancer cells are degenerated. In further or other embodiments, 25% of the cancer cells are degenerated. In further or other embodiments, 30% of the cancer cells are degenerated. In further or other embodiments, 40% of the cancer cells are degenerated. In further or other embodiments, 50% of the cancer cells are degenerated. In further or other embodiments, 60% of the cancer cells are degenerated. In further or other embodiments, 70% of the cancer cells are degenerated. In further or other embodiments, 75% of the cancer cells are degenerated. In further or other embodiments, 80% of the cancer cells are degenerated. In further or other embodiments, 90% of the cancer cells are degenerated. In further or other embodiments, 100% of the cancer cells are degenerated. In further or other embodiments, substantially all of the cancer cells are degenerated. In various embodiments, the above-described degeneration occurs within 1 day, 5 days, 10 days, 1 month, 2 months, 6 months, or a year.

In some embodiments, the cancer cells are killed. In further or other embodiments, 1% of the cancer cells are killed. In further or other embodiments, 2% of the cancer cells are killed. In further or other embodiments, 3% of the cancer cells are killed. In further or other embodiments, 4% of the cancer cells are killed. In further or other embodiments, 5% of the cancer cells are killed. In further or other embodiments, 10% of the cancer cells are killed. In further or other embodiments, 20% of the cancer cells are killed. In further or other embodiments, 25% of the cancer cells are killed. In further or other embodiments, 30% of the cancer cells are killed. In further or other embodiments, 40% of the cancer cells are killed. In further or other embodiments, 50% of the cancer cells are killed. In further or other embodiments, 60% of the cancer cells are killed. In further or other embodiments, 70% of the cancer cells are killed. In further or other embodiments, 75% of the cancer cells are killed. In further or other embodiments, 80% of the cancer cells are killed. In further or other embodiments, 90% of the cancer cells are killed. In further or other embodiments, 100% of the cancer cells are killed. In further or other embodiments, substantially all of the cancer cells are killed. In various embodiments, the killing of cancer cells as described above occurs within 1 day, 5 days, 10 days, 1 month, 2 months, 6 months, or 1 year.

In further or other embodiments, the growth of the cancer cell is inhibited. In further or other embodiments, the growth of the cancer cells is inhibited by about 1%. In further or other embodiments, the growth of the cancer cells is inhibited by about 2%. In further or other embodiments, the growth of the cancer cells is inhibited by about 3%. In further or other embodiments, the growth of the cancer cells is inhibited by about 4%. In further or other embodiments, the growth of the cancer cells is inhibited by about 5%. In further or other embodiments, the growth of the cancer cells is inhibited by about 10%. In further or other embodiments, the growth of the cancer cells is inhibited by about 20%. In further or other embodiments, the growth of the cancer cells is inhibited by about 25%. In further or other embodiments, the growth of the cancer cells is inhibited by about 30%. In further or other embodiments, the growth of the cancer cells is inhibited by about 40%. In further or other embodiments, the growth of the cancer cells is inhibited by about 50%. In further or other embodiments, the growth of the cancer cells is inhibited by about 60%. In further or other embodiments, the growth of the cancer cells is inhibited by about 70%. In further or other embodiments, the growth of the cancer cells is inhibited by about 75%. In further or other embodiments, the growth of the cancer cells is inhibited by about 80%. In further or other embodiments, the growth of the cancer cells is inhibited by about 90%. In further or other embodiments, the growth of the cancer cells is inhibited by about 100%. In various embodiments, the inhibition occurs within 1 day, 5 days, 10 days, 1 month, 2 months, 6 months, or 1 year.

In other aspects, the invention relates to a method of reducing tumor size, inhibiting tumor size increase, reducing tumor proliferation, or preventing tumor proliferation in a subject, comprising administering to the subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof. In some embodiments, the size of the tumor is reduced. In further or other embodiments, the size of the tumor is reduced by at least 1%. In further or other embodiments, the size of the tumor is reduced by at least 2%. In further or other embodiments, the size of the tumor is reduced by at least 3%. In further or other embodiments, the size of the tumor is reduced by at least 4%. In further or other embodiments, the size of the tumor is reduced by at least 5%. In further or other embodiments, the size of the tumor is reduced by at least 10%. In further or other embodiments, the size of the tumor is reduced by at least 20%. In further or other embodiments, the size of the tumor is reduced by at least 25%. In further or other embodiments, the size of the tumor is reduced by at least 30%. In further or other embodiments, the size of the tumor is reduced by at least 40%. In further or other embodiments, the size of the tumor is reduced by at least 50%. In further or other embodiments, the size of the tumor is reduced by at least 60%. In further or other embodiments, the size of the tumor is reduced by at least 70%. In further or other embodiments, the size of the tumor is reduced by at least 75%. In further or other embodiments, the size of the tumor is reduced by at least 80%. In further or other embodiments, the size of the tumor is reduced by at least 85%. In further or other embodiments, the size of the tumor is reduced by at least 90%. In further or other embodiments, the size of the tumor is reduced by at least 95%. In further or other embodiments, the tumor is eradicated. In some embodiments, the size of the tumor is not increased. In various embodiments, the above effect on tumor size occurs within 1 day, 5 days, 10 days, 1 month, 2 months, 6 months, or 1 year.

In some embodiments, tumor proliferation is reduced. In some embodiments, tumor proliferation is reduced by at least 1%. In some embodiments, tumor proliferation is reduced by at least 2%. In some embodiments, tumor proliferation is reduced by at least 3%. In some embodiments, tumor proliferation is reduced by at least 4%. In some embodiments, tumor proliferation is reduced by at least 5%. In some embodiments, tumor proliferation is reduced by at least 10%. In some embodiments, tumor proliferation is reduced by at least 20%. In some embodiments, tumor proliferation is reduced by at least 25%. In some embodiments, tumor proliferation is reduced by at least 30%. In some embodiments, tumor proliferation is reduced by at least 40%. In some embodiments, tumor proliferation is reduced by at least 50%. In some embodiments, tumor proliferation is reduced by at least 60%. In some embodiments, tumor proliferation is reduced by at least 70%. In some embodiments, tumor proliferation is reduced by at least 75%. In some embodiments, tumor proliferation is reduced by at least 75%. In some embodiments, tumor proliferation is reduced by at least 80%. In some embodiments, tumor proliferation is reduced by at least 90%. In some embodiments, tumor proliferation is reduced by at least 95%. In some embodiments, tumor proliferation is prevented. In various embodiments, the above-described effect on cell proliferation occurs within 1 day, 5 days, 10 days, 1 month, 2 months, 6 months, or 1 year.

In some embodiments, the composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins; antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormone/anti-hormone therapeutics and hematopoietic growth factors. In further or other embodiments, the therapeutic agent is selected from paclitaxel, bortezomib, or both.

Proliferative diseases

In other aspects, the invention relates to treating or preventing increase in an individualCulture mediumA method of treating sexual disorders, comprising administering to the subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In some embodiments, the increase is Culture mediumThe sexual disease is cancer, psoriasis, restenosis, autoimmunityDisease or atherosclerosis. In further or other embodiments, the increaseCulture mediumThe sexual disorder is a hyperproliferative disorder. In further or other embodiments, the increaseCulture mediumThe sexual disorder is selected from the group consisting of a tumor, leukemia, a neoplasm, a cancer, a carcinoma, and a malignant disease. In further or other embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, colorectal cancer, gastric cancer, head and neck cancer, or leukemia. In further or other embodiments, the fibrotic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid formation, interstitial nephritis, or pulmonary fibrosis. In further or other embodiments, the cancer is gastric cancer, brain cancer, breast cancer, lung cancer, non-small cell lung cancer, ovarian cancer, pancreatic cancer, liver cancer, prostate cancer, renal cancer, colorectal cancer, or leukemia. In further or other embodiments, the cancer is brain cancer or adrenocortical cancer. In further or other embodiments, the cancer is breast cancer. In further or other embodiments, the cancer is ovarian cancer. In further or other embodiments, the cancer is pancreatic cancer. In further or other embodiments, the cancer is prostate cancer. In further or other embodiments, the cancer is renal cancer. In further or other embodiments, the cancer is colorectal cancer. In further or other embodiments, the cancer is myeloid leukemia. In further or other embodiments, the cancer is glioblastoma. In further or other embodiments, the cancer is follicular lymphoma. In further or other embodiments, the cancer is pre-B acute leukemia. In further or other embodiments, the cancer is chronic lymphocytic B leukemia. In further or other embodiments, the cancer is mesothelioma. In further or other embodiments, the cancer is small cell lung cancer. In other embodiments, the cancer is gastric cancer.

In some embodiments, the combination comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins; antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormone/anti-hormone therapeutics and hematopoietic growth factors.

In further or other embodiments, the therapeutic agent is selected from paclitaxel, bortezomib, or both. In some embodiments, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly, or by infusion), topically, or rectally.

In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the subject having a proliferative disease is a mammal. In further or other embodiments, the subject is a human. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

Inflammatory diseases

In other aspects, the invention relates to a method of treating or preventing an inflammatory disease in a subject, comprising administering to the subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In further or other embodiments, the inflammatory disease is selected from chronic inflammatory disease, rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, juvenile arthritis, acute rheumatic arthritis, enteropathic arthritis, neuropathic arthritis, psoriatic arthritis, suppurative arthritis, atherosclerosis, systemic lupus erythematosus, inflammatory bowel disease, irritable bowel syndrome, ulcerative colitis, reflux esophagitis, crohn's disease, gastritis, asthma, allergy, respiratory distress syndrome, pancreatitis, chronic obstructive pulmonary disease, pulmonary fibrosis, psoriasis, eczema, or scleroderma.

In some embodiments, a composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent.

In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the subject having an inflammatory disease is a mammal. In further or other embodiments, the subject is a human. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

Cancer treatment

In other aspects, the invention relates to methods of treating or preventing cancer in a subject comprising administering to the subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof. In further or other embodiments, the cancer is brain cancer, breast cancer, gastric cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, colorectal cancer, or leukemia. In further or other embodiments, the fibrotic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid formation, interstitial nephritis, or pulmonary fibrosis. In further or other embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, gastric cancer, colorectal cancer, or leukemia. In further or other embodiments, the cancer is brain cancer or adrenocortical cancer. In further or other embodiments, the cancer is breast cancer. In further or other embodiments, the cancer is ovarian cancer. In further or other embodiments, the cancer is pancreatic cancer. In further or other embodiments, the cancer is prostate cancer. In further or other embodiments, the cancer is renal cancer. In further or other embodiments, the cancer is colorectal cancer. In further or other embodiments, the cancer is myeloid leukemia. In further or other embodiments, the cancer is glioblastoma. In further or other embodiments, the cancer is follicular lymphoma. In further or other embodiments, the cancer is pre-B acute leukemia. In further or other embodiments, the cancer is chronic lymphocytic B leukemia. In further or other embodiments, the cancer is mesothelioma. In further or other embodiments, the cancer is small cell lung cancer. In some embodiments, the cancer is gastric cancer.

In some embodiments, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly, or by infusion), topically, or rectally. In further or other embodiments, the amount of the compound of formula I ranges from about 0.001 to about 1000mg/kg body weight/day. In further or other embodiments, the amount of the compound of formula I is in the range of about 0.5 to about 50 mg/kg/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.001 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.01 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.02 to about 5 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.05 to about 2.5 g/day. In further or other embodiments, the amount of the compound of formula I is from about 0.1 to about 1 g/day. In further or alternative embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate.

In further or alternative embodiments, dosage levels above the upper limit of the range recited above may be necessary. In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the individual having cancer is a mammal. In further or other embodiments, the subject is a human. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

Introduction by reference

All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

Brief Description of Drawings

The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings.

FIG. 1 shows: graph of mean tumor volume versus time (days) in mice implanted with A375 melanoma, Colo205 colon tumor, A431 epidermoid tumor, or HT-29 colon tumor cells. Mice were orally administered once daily (25mg/kg, 50mg/kg or 100mg/kg) for 14 days.

FIG. 2 shows: a% tumor growth inhibition (% TGI) profile in A375 xenografted mice dosed with 50mg/kg QD, 25mg/kg BID, 50mg/kg QD, and 12.5mg/kg BID.

FIG. 3 shows: graph of plasma concentration (log nM) versus pERK% inhibition in female nu/nu mice implanted with Colo205 tumor cells. Mice were administered single doses of 2.5, 5, 10 or 25 mg/kg.

FIG. 4 shows: graph of plasma concentration (ng/mL) versus time (hours) in humans after single dose administration of 2mg (2x1mg capsules), 4mg (4x1mg capsules), or 6mg (6x1mg capsules).

FIG. 5 is a powder X-ray diffraction (PXRD) pattern of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, form A, produced using an Inel XRG-3000 diffractometer. The peak intensity, defined by counts per second, is plotted against diffraction angle 2 θ (degrees).

FIG. 6 is a modulated Differential Scanning Calorimetry (DSC) thermogram of form A N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide produced using a TA instruments differential scanning calorimeter Q1000. Normalized heat flow in watts/gram (W/g) is plotted against measured sample temperature in degrees celsius.

FIG. 7 is a diagram of: PXRD pattern of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide form a (top) and PXRD pattern of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide amorphous (bottom) produced using Inel XRG-3000 diffractometer. The peak intensity, defined by counts per second, is plotted against diffraction angle 2 θ (degrees).

Fig. 8 shows: dynamic vapor adsorption/Desorption (DVS) isotherms of the N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide type a produced using a VTI SGA-100 vapor adsorption analyzer.

Fig. 9 shows: thermogravimetric (TG) thermogram of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (a) type generated using TA instruments 2950 thermogravimetric analyzer.

Fig. 10(a) and 10(b) show: growth arrest of dividing A375 cells in log phase in contact with increasing concentrations of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. The assay cells were assayed for ATP content. 100% growth arrest was determined using 1 μ M N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Fig. 11 shows: 48hr AK assay in A375 cells. Log phase dividing a375 cells were contacted with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and PD-325901 for 48hr and analyzed for AK release.

FIGS. 12A-12C show: n- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (A) inhibits Colo205 cell Growth (GI) in human colorectal cancer₅₀=11 nM); (B) inhibition of A375 cell Growth (GI)₅₀=22nM), and (C) inhibits MDA-MB231 cells, which show no growth arrest by N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide in a two-dimensional adherence dependent assay.

FIG. 13A shows Colo205 cell growth inhibition in human colorectal cancer, GI₅₀Values were 6nM and 11nM, respectively.

FIG. 13B shows inhibition of A375 cell growth, GI₅₀Values were 5nM and 22 nM.

Fig. 14A and 14B show: the effect of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on cell cycle progression demonstrates that contacting a375 cells with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide causes arrest in the G1 phase of the cell cycle, as indicated by a decrease in cells in both G2 and S phases.

Fig. 15A and 15B show: effect of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on gastric cancer (gastric adenocarcinoma) cell line AGS after 3 days (fig. 15A) and after 6 days (fig. 15B). The y-axis is the number of cells relative to the carrier and the x-axis is the concentration uM of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Fig. 16 shows: average liver weight in tumor-bearing mice after treatment with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (2mg/kg, once daily, oral; 10mg/kg, once daily, oral; and 50mg/kg, once daily, oral).

Fig. 17 shows: liver tumor weight in tumor-bearing mice after treatment with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (2mg/kg, once daily, oral; 10mg/kg, once daily, oral; and 50mg/kg, once daily, oral).

Fig. 18 shows: mean tumor weight after treatment with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (2 mg/kg; 10mg/kg; and 50 mg/kg).

Fig. 19 shows: hs746t inhibition of cell proliferation in a plot of cell number (relative to vehicle) versus N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide concentration.

Fig. 20A shows: a graph comparing the levels of apoptosis with increasing concentrations of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of non-small cell lung cancer (NSCLC) MV522 cells treatment.

Fig. 20B shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of non-small cell lung cancer (NSCLC) H358 cells treatment.

Fig. 20C shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 6 of non-small cell lung cancer (NSCLC) a549 cell treatment.

Fig. 20D shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of non-small cell lung cancer (NSCLC) H727 cell treatment.

Fig. 20E shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of colon HT29 cells.

Fig. 20F shows: graph of the levels of apoptosis increased with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 6 of treatment of colonic HCT116 cells.

Fig. 20G shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of colon HUH7 hepatoma cells.

Fig. 20H shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of sarcoma U2-OS cells treatment.

Fig. 20I shows: graph of the levels of apoptosis increasing with the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of glioma D37 cells.

Fig. 21 shows: compound a was selective for MEK1 and MEK2 versus a panel of 205 enzymes at 10 μ M. The cell lines are Colo205, A375, A431 and HT-29.

Fig. 22 shows: graph of paw volume increase and edema reduction relative to vehicle control in each treatment group after administration of 6, 20, 60 and 200mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide to rats in the rat carrageenan paw edema model.

Fig. 23A shows: inhibition of swelling in an adjuvant-induced arthritis model in the acute phase in rats treated with 2,6 and 20mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Fig. 23B shows: inhibition of swelling in an adjuvant-induced arthritis model in the delayed phase in rats treated with 2, 6 and 20mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Fig. 24 shows: mean arthritis score in collagen-antibody induced arthritis (CAIA) mice treated with 1, 3&10mg/kg QD N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Detailed Description

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All or a portion of the documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are hereby expressly incorporated by reference in their entirety for any purpose.

Certain chemical terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, published materials referred to throughout this disclosure are incorporated herein by reference in their entirety, unless otherwise indicated. If there are multiple definitions of terms in this document, the definitions in this section prevail. In the case of references to URLs or other such identifiers or addresses, it is understood that such identifiers may change and that the specific information on the internet may change constantly, but equivalent information may be found by searching the internet or other suitable reference sources. The reference demonstrates the availability and public dissemination of such information.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or" means "and/or" unless stated otherwise. Moreover, the use of the term "including" and other forms is not limiting.

Definitions of the standardized chemical terms can be found in the reference works (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY, 4 th edition" volume A (2000) and volume B (2001), Plenum Press, New York). Unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, IR and UV/Vis spectroscopy and pharmacology are used in the art. Unless a specific definition is provided, the nomenclature used in connection with the analytical chemistry, synthetic organic chemistry, and pharmaceutical chemistry described herein, and the laboratory procedures and techniques in these areas, are those known in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and administration, and treatment of patients. For example, reaction and purification techniques can be performed using manufacturer kit instructions, or in a manner commonly done in the art, or as described herein. The techniques and methods described above can generally be performed by conventional methods well known in the art, and as described in various general and more specific references that are cited and discussed throughout the present specification. Throughout this specification, one skilled in the art will be able to select groups and substituents thereof to provide stable moieties and compounds.

Where substituent groups are illustrated by their conventional chemical formula written from left to right, they likewise include chemically identical substituents that would result from writing the structure from right to left. with-CH₂O-corresponds to-OCH₂As non-limiting examples.

Unless otherwise indicated, the use of general chemical terms such as, but not limited to, "alkyl", "amine", "aryl" correspond to their optionally substituted forms. For example, "alkyl" as used herein includes optionally substituted alkyl.

The compounds described herein may have one or more stereocenters, and each center may exist in the R or S configuration, or a combination thereof. Likewise, the compounds described herein may have one or more double bonds, and each double bond may be present in the E (trans) or Z (cis) configuration, or a combination thereof. It is understood that the description of a particular stereoisomer, regioisomer (regioisomer), diastereomer, enantiomer or epimer includes all possible stereoisomers, regioisomers, diastereomers, enantiomers or epimers and mixtures thereof. Thus, the compounds described herein include all individual configurational stereoisomeric forms, positional stereoisomeric forms, diastereomeric forms, enantiomeric forms and epimeric forms, and mixtures thereof. It is understood that the description of a particular chemical structure or chemical name for a compound containing one or more chiral centers without specifying a particular stereochemistry includes all possible stereoisomers, including mixtures of all possible stereoisomers, pure or substantially pure forms of a particular stereoisomer, and pure or substantially pure forms of alternative stereoisomers. Techniques for inverting or leaving unchanged a particular stereocenter, as well as techniques for resolving stereoisomers, are well known in the art, and it is well within the ability of those skilled in the art to select an appropriate method for a particular situation. See, e.g., Furniss et al (eds.), VOGEL' S ENCYCLOPEDIA OF practicalalanic CHEMISTRY 5. th ed., Longman Scientific and Technical ltd., Essex,1991, 809-816; and Heller, acc, chem, res, 1990,23,128.

The terms "moiety," "chemical moiety," "group," and "chemical group" as used herein mean a specific moiety or functional group of a molecule. Chemical moieties are generally considered to be chemical entities embedded within or attached to a molecule.

The term "bond" or "single bond" means a chemical bond between two atoms, or, when the atoms connected by the bond are considered part of a larger substructure, between two parts.

The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" means "alkyl" or "substituted alkyl" as defined below. Further, the optionally substituted group may be unsubstituted (e.g., -CH)₂CH₃) Fully substituted (e.g. -CF)₂CF₃) Monosubstituted (e.g. -CH)₂CH₂F) Or any level of substitution between fully and mono-substituted (e.g., -CH)₂CHF₂、-CH₂CF₃、-CF₂CH₃、-CFHCHF₂Etc.). For any group containing one or more substituents, those skilled in the art will appreciate that such groups are not intended to introduce any substitution or substitution pattern that is not sterically impractical and/or synthetically non-feasible (e.g., substituted alkyl includes optionally substituted cycloalkyl groups, which in turn are defined as including optionally substituted alkyl groups, possibly infinite). Thus, any substituent described should generally be understood as having a maximum molecular weight of about 1,000 daltons, more typically up to about 500 daltons (except in those cases where macromolecular substituents such as polypeptides, polysaccharides, polyethylene glycols, DNA, RNA, and the like are expressly intended).

Unless otherwise indicated, the use of general chemical terms such as, but not limited to, "alkyl", "amine", "aryl" is unsubstituted.

As used herein C₁-C_xComprising C₁-C₂、C₁-C₃……C₁-C_x. By way of example only, denoted as "C₁-C₄"indicates the presence of 1 to 4 carbon atoms in the moiety, i.e., containing 1 carbon atom, 2 carbon atoms, 3 carbon atoms, or 4 carbon atoms, and range C₁-C₂And C₁-C₃A group of (1). Thus, by way of example only, "C₁-C₄Alkyl "indicates the presence of 1 to 4 carbon atoms in the alkyl group, i.e., the alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Whenever a numerical range such as "1 to 10" appears herein, each integer within the given range is meant; for example, "1 to 10 carbon atoms" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

The term "a and a' together with the carbon atom to which they are attached form a 3-6 membered saturated ring" as used herein means the following structure of the compound of formula I:

the term "heteroatom" or "hetero", as used herein, alone or in combination, means an atom other than carbon or hydrogen. The heteroatoms may be independently selected from oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin, but are not limited to these atoms. In embodiments where two or more heteroatoms are present, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may each be different from the other heteroatoms.

The term "alkyl" as used herein, alone or in combination, is intended to encompassA linear or branched saturated hydrocarbon monovalent radical of 1 to about 10 carbon atoms, or 1-6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2-dimethyl-1-butyl, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1, N-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl and hexyl, and longer alkyl groups such as heptyl, octyl and the like. Whenever a range of values, e.g. "C₁-C₆Alkyl "or" C_1-6Alkyl "when present herein means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. In one embodiment, the "alkyl" is substituted. Unless otherwise indicated, the "alkyl" is unsubstituted.

The term "alkenyl", as used herein, alone or in combination, means a straight or branched chain hydrocarbon monovalent radical having one or more carbon-carbon double bonds and having from 2 to about 10 carbon atoms, or from 2 to about 6 carbon atoms. The groups may be in either the cis or trans configuration for the double bond, and it is understood that the groups include both isomers. Examples include, but are not limited to, vinyl (-CH = CH)₂) 1-propenyl (-CH)₂CH=CH₂) Isopropenyl [ -C (CH)₃)=CH₂]Butenyl, 1, 3-butadienyl and the like. Whenever a range of values, e.g. "C₂-C₆Alkenyl "or" C_2-6Alkenyl "when present herein means that the alkenyl group can consist of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. In one embodiment, the "alkenyl" group is substituted. Unless otherwise indicated, the "alkenyl" is unsubstituted.

The term "alkynyl", as used herein, alone or in combination, means having one or more carbon-carbon triple bonds andand having a linear or branched hydrocarbon monovalent radical of 2 to about 10 carbon atoms, or 2 to about 6 carbon atoms. Examples include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, 1, 3-butadiynyl, and the like. Whenever a range of values, e.g. "C ₂-C₆Alkynyl "or" C_2-6Alkynyl "when present herein means that the alkynyl group can consist of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. In one embodiment, the "alkynyl" group is substituted. Unless otherwise indicated, the "alkynyl" is unsubstituted.

The terms "heteroalkyl," "heteroalkenyl," and "heteroalkynyl" as used herein, alone or in combination, mean alkyl, alkenyl, and alkynyl structures, respectively, as described above, wherein one or more backbone chain carbon atoms (and any associated appropriate hydrogen atoms) are each independently heteroatom (i.e., an atom other than carbon, such as, but not limited to, oxygen, nitrogen, sulfur, silicon, phosphorus, tin, or combinations thereof) or heteroatomic group (such as, but not limited to, -O-, -S-, -O-S-, -S-O-, ═ N-N ═ N-, -N ═ N-NH-, -p (O))₂-、-O-P(O)₂-、-P(O)₂-O-、-S(O)-、-S(O)₂-、-SnH₂-etc.) substitution.

The terms "haloalkyl", "haloalkenyl" and "haloalkynyl" as used herein, alone or in combination, mean alkyl, alkenyl and alkynyl groups, respectively, as defined above, wherein one or more hydrogen atoms are replaced by a fluorine atom, a chlorine atom, a bromine atom or an iodine atom, or combinations thereof. In some embodiments, two or more hydrogen atoms may be substituted with the same halogen atom as each other (e.g., difluoromethyl); in other embodiments, two or more hydrogen atoms may be substituted with halogen atoms that are not all identical to each other (e.g., 1-chloro-1-fluoro-1-iodoethyl). Non-limiting examples of haloalkyl groups are fluoromethyl, chloromethyl and bromoethyl. A non-limiting example of a haloalkenyl group is bromovinyl. A non-limiting example of a haloalkynyl group is chloroethynyl.

The term "carbon chain" as used herein, alone or in combination, means any alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl or heteroalkynyl group, which is linear, cyclic, or any combination thereof. If the chain is part of a linker that contains one or more rings as part of the core backbone, for purposes of calculating chain length, the "chain" includes only those carbon atoms that make up the bottom or top of a given ring, rather than both the bottom and top, and where the top and bottom of the ring are not equal in length, a shorter distance should be used to determine chain length. If the chain contains heteroatoms as part of the backbone, those atoms are not counted as part of the carbon chain length.

The terms "ring", "cyclic", "ring (ring) and" membered ring "as used herein, alone or in combination, mean any covalently closed structure, including alicyclic, heterocyclic, aromatic, heteroaromatic and polycyclic fused or non-fused ring systems as described herein. The ring may be optionally substituted. The rings may form part of a fused ring system. The term "element" means the number of backbone atoms that make up the ring. Thus, by way of example only, cyclohexane, pyridine, pyran and pyrimidine are six-membered rings, while cyclopentane, pyrrole, tetrahydrofuran and thiophene are five-membered rings.

The term "fused" as used herein, alone or in combination, means a ring structure in which two or more rings share one or more bonds.

The term "cycloalkyl", as used herein, alone or in combination, means a saturated hydrocarbon monovalent ring containing from 3 to about 15 ring carbon atoms or from 3 to about 10 ring carbon atoms, but may include as substituents additional acyclic carbon atoms (e.g., methylcyclopropyl). Whenever a range of values, e.g. "C₃-C₆Cycloalkyl radicals "or" C_3-6Cycloalkyl "when present herein means that the cycloalkyl group may consist of 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms, i.e. cyclopropyl, cyclobutyl, cyclopentyl or cycloheptyl (cyclohepty), although the present definition also covers the occurrence of the term" cycloalkyl "where no numerical range is specified. This term includes fused, non-fused, bridge and spiro groups. The fused cycloalkyl group mayTo contain 2-4 fused rings, wherein the connecting ring is a cycloalkyl ring and each of the other rings can be alicyclic, heterocyclic, aromatic or heteroaromatic or any combination thereof. Examples include, but are not limited to, cyclopropyl, cyclopentyl, cyclohexyl, decahydronaphthyl, and bicyclo [2.2.1]Heptyl and adamantyl ring systems. Illustrative examples include, but are not limited to, the following:

In one embodiment, the "cycloalkyl" is substituted. Unless otherwise indicated, the "cycloalkyl" is unsubstituted.

The terms "non-aromatic heterocyclyl" and "heteroalicyclic" as used herein, alone or in combination, mean a monovalent radical of a saturated, partially unsaturated, or fully unsaturated non-aromatic ring containing from 3 to about 20 ring atoms, wherein one or more of the ring atoms is an atom other than carbon independently selected from the group consisting of, but not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin. In embodiments where two or more heteroatoms are present within a ring, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may each be different from the other heteroatoms. These terms include fused, non-fused, bridge and spiro groups. The fused non-aromatic heterocyclyl group may contain 2 to 4 fused rings, where the connecting ring is a non-aromatic heterocyclic ring and the other rings may be alicyclic, heterocyclic, aromatic or heteroaromatic rings or any combination thereof. Fused ring systems may be fused via single or double bonds, and may also be fused via carbon-carbon, carbon-heteroatom or heteroatom-heteroatom bonds. These terms also include groups having from 3 to about 12 skeletal ring atoms and having from 3 to about 10 skeletal ring atoms. The attachment of the non-aromatic heterocyclic subunit to its parent molecule may be through a heteroatom or a carbon atom. Likewise, other substitutions may be through heteroatoms or carbon atoms. By way of non-limiting example, the imidazolidine non-aromatic heterocycle may be substituted by any of its N atoms (II) Imidazolidin-1-yl or imidazolidin-3-yl) or any of its carbon atoms (imidazolidin-2-yl, imidazolidin-4-yl or imidazolidin-5-yl) is attached to the parent molecule. In certain embodiments, the non-aromatic heterocyclic ring comprises one or more carbonyl or thiocarbonyl groups such as oxo-and thio-containing groups. Examples include, but are not limited to, pyrrolidinyl, tetrahydrofuryl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepanyl, and the likeRadical diazaRadical, sulfur nitrogen heteroA group, 1,2,3, 6-tetrahydropyridinyl group, 2-pyrrolinyl group, 3-pyrrolinyl group, indolinyl group, 2H-pyranyl group, 4H-pyranyl group, dioxanyl group, 1, 3-dioxolanyl group, pyrazolinyl group, dithianyl group, dithiolanyl group, dihydropyranyl group, dihydrothienyl group, dihydrofuranyl group, pyrazolidinyl group, imidazolinyl group, imidazolidinyl group, 3-azabicyclo [3.1.0]Hexyl, 3-azabicyclo [4.1.0]Heptyl, 3H-indolyl and quinolizinyl. Illustrative examples of heterocycloalkyl groups, also referred to as non-aromatic heterocyclyl groups, include:

The term also includes all ring forms of carbohydrates including, but not limited to, monosaccharides, disaccharides, and oligosaccharides. In one embodiment, the "non-aromatic heterocyclyl" or "heteroalicyclic" group is substituted. Unless otherwise indicated, the "non-aromatic heterocyclyl" or "heteroalicyclic" group is unsubstituted.

The term "aryl" as used herein, alone or in combination, means an aromatic hydrocarbon radical of from 6 to about 20 ring carbon atoms, and includes fused and non-fused aryl rings. Fused aryl ring groups contain 2-4 fused rings, where the connecting ring is an aryl ring, and each of the other rings can be alicyclic, heterocyclic, aromatic or heteroaromatic, or any combination thereof. In addition, the term aryl includes fused and non-fused rings containing from 6 to about 12 ring carbon atoms and from 6 to about 10 ring carbon atoms. Non-limiting examples of monocyclic aryl groups include phenyl; non-limiting examples of fused ring aryl groups include naphthyl, phenanthryl, anthracenyl, azulenyl; non-limiting examples of non-fused biaryl groups include biphenyl. In one embodiment, the "aryl" is substituted. Unless otherwise indicated, the "aryl" is unsubstituted.

The term "heteroaryl" as used herein, alone or in combination, means an aromatic monovalent radical comprising from about 5 to about 20 skeletal ring atoms, wherein one or more of the ring atoms is a heteroatom independently selected from the group consisting of, but not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin, with the proviso that the ring of the radical does not contain two adjacent O or S atoms. In embodiments where two or more heteroatoms are present within a ring, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may each be different from the other heteroatoms. The term heteroaryl includes fused and non-fused heteroaryl groups having at least one heteroatom. The term heteroaryl also includes fused and non-fused heteroaryl groups having from 5 to about 12 skeletal ring atoms and having from 5 to about 10 skeletal ring atoms. The bond to the heteroaryl group may be through a carbon atom or a heteroatom. Thus, as a non-limiting example, an imidazole group may be attached to the parent molecule through any of its carbon atoms (imidazol-2-yl, imidazol-4-yl, or imidazol-5-yl), or any of its nitrogen atoms (imidazol-1-yl or imidazol-3-yl). Likewise, a heteroaryl group may be further substituted by any or all of its carbon atoms and/or any or all of its heteroatoms. The fused heteroaryl group can contain 2 to 4 fused rings, wherein the connecting ring is a heteroaromatic ring and the other rings can be alicyclic, heterocyclic, aromatic, heteroaromatic or any combination thereof. Non-limiting examples of monocyclic heteroaryl groups include pyridyl; non-limiting examples of fused ring heteroaryl groups include benzimidazolyl, quinolinyl, acridinyl; non-limiting examples of non-fused biaryl groups include bipyridyl. Other examples of heteroaryl groups include, without limitation, furyl, thienyl, oxazolyl, acridinyl, phenazinyl, benzimidazolyl, benzofuryl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzothiophenyl, benzooxadiazolyl, benzotriazolyl, imidazolyl, indolyl, isoxazolyl, isoquinolyl, indolizinyl, isothiazolyl, isoindolyl oxadiazolyl, indazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrrolyl, pyrazinyl, pyrazolyl, purinyl, phthalazinyl, pteridinyl, quinolinyl, quinazolinyl, quinoxalinyl, triazolyl, tetrazolyl, thiazolyl, triazinyl, thiadiazolyl, and the like, and oxides thereof, such as pyridyl-N-oxide. Illustrative examples of heteroaryl groups include the following moieties:

In one embodiment, the "heteroaryl" is substituted. Unless otherwise indicated, the "heteroaryl" is unsubstituted.

The term "heterocyclyl", as used herein, alone or in combination, means both heteroalicyclic and heteroaryl groups. Herein, whenever the number of carbon atoms in the heterocycle (e.g. C) is specified₁-C₆Heterocyclic ring), at least one non-carbon atom (heteroatom) must be present within the ring. Names such as "C₁-C₆The heterocyclic ring "means only the number of carbon atoms in the ring and does not mean the total number of atoms in the ring. By a name such as "4-6 membered heterocyclic" is meant the total number of atoms contained within the ring (i.e., a 4-membered, 5-membered, or 6-membered ring in which at least one atom is a carbon atom, at leastOne atom is a heteroatom and the remaining 2-4 atoms are carbon or heteroatoms). For heterocycles having two or more heteroatoms, those two or more heteroatoms may be the same or different from each other. Non-aromatic heterocyclic groups include groups having only 3 atoms in the ring, whereas aromatic heterocyclic groups must have at least 5 atoms in the ring. May be bonded to the heterocycle (i.e., attached to the parent molecule or further substituted) through a heteroatom or carbon atom. In one embodiment, the "heterocyclyl" is substituted. Unless otherwise indicated, the "heterocyclyl" is unsubstituted.

The terms "halogen", "halo" or "halide", as used herein, alone or in combination, mean fluorine, chlorine, bromine and/or iodine.

The term "amino" as used herein, alone or in combination, means a monovalent radical-NH₂。

The term "alkylamino" as used herein, alone or in combination, means the monovalent radical-NH (alkyl), wherein alkyl is as defined herein.

The term "dialkylamino," as used herein, alone or in combination, means a monovalent radical-N (alkyl), wherein each alkyl can be the same or different and is as defined herein.

The term "diaminoalkyl" as used herein, alone or in combination, means an alkyl group comprising two amine groups, wherein the amine groups may be substituents on the alkyl group, which may be amino, alkylamino or dialkylamino groups, or wherein one or two of the amine groups may constitute part of the alkyl chain to form-alkylene-N (H or alkyl) -alkylene-N (H or alkyl or alkylene-).

The term "hydroxy" as used herein, alone or in combination, means a monovalent radical-OH.

The term "cyano," as used herein, alone or in combination, means a monovalent radical — CN.

Terms used herein, alone or in combination " Cyanomethyl "means a monovalent radical-CH₂CN。

The term "nitro" as used herein, alone or in combination, means a monovalent radical-NO₂。

The term "oxy", as used herein alone or in combination, means the divalent radical-O-.

The term "oxo" as used herein, alone or in combination, means a divalent radical = O.

The term "carbonyl", as used herein, alone or in combination, means a divalent radical-C (= O) -, which may also be written as-C (O) -.

The term "carboxy" as used herein, alone or in combination, means a-C (O) OH moiety, which may also be written as-COOH.

The term "alkoxy" as used herein, alone or in combination, means an alkyl ether group, -O-alkyl, which includes the groups-O-aliphatic and-O-carbocyclyl, wherein the alkyl, aliphatic and carbocyclyl groups may be optionally substituted, and wherein the terms alkyl, aliphatic and carbocyclyl are as defined herein. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like.

The term "sulfinyl", as used herein, alone or in combination, means the divalent radical-S (= O) -.

The term "sulfonyl", as used herein, alone or in combination, means a divalent radical-S (= O) ₂-。

The terms "sulfonamide", "sulfonamido", and "aminosulfonyl" (sulfamoyl) as used herein, alone or in combination, mean the diradical radical-S (= O)₂-NH-and-NH-S (= O)₂-。

The terms "sulfamide" and "aminosulfonylamino" as used herein, alone or in combination, mean the diradical radical-NH-S (= O)₂-NH-。

The term "reactant" as used herein means a nucleophile or electrophile used to generate a covalent bond.

It is understood that where two or more groups are used in succession to define a substituent attached to a structure, the first named group is considered to be terminal and the last named group is considered to be attached to the structure in question. Thus, for example, the group arylalkyl is linked to the structure in question via an alkyl group.

Certain pharmaceutical terms

The term "MEK inhibitor" as used herein means that it exhibits an IC of no more than about 100 μ M or no more than about 50 μ M for MEK activity as measured by the MEK1 kinase assay outlined herein₅₀The compound of (1). ' IC₅₀"is the concentration of inhibitor that reduces the activity of an enzyme (e.g., MEK) to a level of half maximal. It has been found that the compounds described herein exhibit inhibitory effects on MEK. The compounds of the invention preferably exhibit an IC for MEK of up to about 10 μ M as measured by the Mek1 kinase assay described herein ₅₀More preferably up to about 5. mu.M, even more preferably no more than about 1. mu.M, and most preferably no more than about 200 nM.

The terms "subject", "patient" or "individual" and the like as used herein with respect to a diseased individual include mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class mammalia: humans, non-human primates such as chimpanzees, and other ape and monkey species; livestock such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; including rodents such as rats, mice and guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish, and the like. In one embodiment of the methods and compositions provided herein, the mammal is a human.

The term "treat," "treating," and other grammatical equivalents as used herein includes alleviating, or ameliorating a symptom of a disease or condition, preventing other symptoms, ameliorating, or preventing an underlying metabolic cause of a symptom, inhibiting a disease or condition, e.g., arresting the development of a disease or condition, alleviating a disease or condition, causing regression of a disease or condition, alleviating a condition caused by a disease or condition, or terminating a symptom of a disease or condition, and is intended to include preventing. The term also includes achieving a therapeutic benefit and/or a prophylactic benefit. Therapeutic benefit means eradication or amelioration of the underlying disorder being treated. In addition, improvements are observed in patients who may still be afflicted with an underlying condition by achieving a therapeutic benefit by eradicating or ameliorating one or more physiological symptoms associated with the underlying condition. For prophylactic benefit, the composition can be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even if a diagnosis of the disease has not yet been made.

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein means an amount of at least one agent or compound administered sufficient to treat or prevent a particular disease or condition. The result can be a reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein that is required to provide clinically significant relief from the disease. The appropriate "effective" amount in any individual case can be determined using techniques such as dose escalation studies.

The terms "substantially anhydrous" and "substantially solvent-free" as used herein mean that the crystalline polymorph contains less than 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, or 2% by weight of water or solvent, respectively.

The term "substantially the same" as used herein means a powder X-ray diffraction pattern or a differential scanning calorimetry pattern which may not be identical to the patterns described herein, but which is within the limits of experimental error considered by those skilled in the art.

The term "administering" or the like as used herein means a method that can be used to enable delivery of a compound or composition to a desired site of biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion administration), topical administration and rectal administration. Those skilled in The art are familiar with The administration techniques that may be used with The compounds and methods described herein, such as, for example, Goodman and Gilman, The pharmaceutical Basis of Therapeutics, current edition; pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions described herein are administered orally.

The term "acceptable" as used herein, in terms of a formulation, composition, or ingredient, means that there is no lasting deleterious effect on the overall health of the subject being treated.

The term "pharmaceutically acceptable" as used herein means a material, such as a carrier or diluent, that does not eliminate the biological activity or properties of the compounds described herein and is relatively non-toxic, i.e., the material can be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of a composition in which it is contained.

The term "pharmaceutical composition" as used herein means a biologically active compound, optionally mixed with at least one pharmaceutically acceptable chemical ingredient (such as, but not limited to, carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and/or excipients).

The term "carrier" as used herein means a relatively non-toxic chemical compound or agent that promotes the incorporation of a compound into a cell or tissue.

The term "agonist" as used herein means a molecule such as a compound, drug, enzyme activator, or hormone modulator that enhances the activity of another molecule or the activity of a receptor site.

The term "antagonist" as used herein means a molecule, such as a compound, drug, enzyme inhibitor, or hormone modulator, that reduces or prevents the action of another molecule or the activity of a receptor site.

The term "modulate" as used herein means to interact with a target, either directly or indirectly, to alter the activity of the target, including, by way of example only, to enhance the activity of the target, inhibit the activity of the target, limit the activity of the target, or prolong the activity of the target.

The term "modulator" as used herein means a molecule that interacts directly or indirectly with a target. Such interactions include, but are not limited to, agonist and antagonist interactions.

The term "pharmaceutically acceptable derivative or prodrug" as used herein means any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of formula I which, when administered to a recipient, is capable of providing, directly or indirectly, a compound of the invention or a pharmaceutically active metabolite or residue thereof. Particularly preferred derivatives or prodrugs are those that enhance the bioavailability of the compounds of the invention when such compounds are administered to a patient (e.g., by allowing an orally administered compound to be more readily absorbed into the blood), or which improve delivery of the parent compound to a biological compartment, such as the brain or lymphatic system.

As used herein, a "prodrug" is a compound that can be converted under physiological conditions or by solvolysis to the specified compound or a pharmaceutically acceptable salt of such a compound. Prodrugs include compounds wherein an amino acid residue or a polypeptide chain of two or more amino acid residues is covalently linked via an amide or ester bond to a free amino, hydroxyl or carboxylic acid group of a compound of formula I. Contemplated amino acid residues include, but are not limited to, the 20 naturally occurring amino acids. Other suitable amino acids include 4-hydroxyproline, hydroxylysine, desmosine (desmosine), isodesmosine (isodemosine), 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, citrulline (cirtuline), homocysteine, homoserine, ornithine and methionine sulfone. Other types of prodrugs are known in the art.

Pharmaceutically acceptable prodrugs of the compounds described herein include, but are not limited to, esters, carbonates, thiocarbonates, N-acyl derivatives, N-acyloxyalkyl derivatives, quaternary ammonium derivatives of tertiary amines, N-Mannich bases, Schiff bases, amino acid conjugates, phosphates, metal salts, and sulfonates. Various forms of prodrugs are known in the art. See, e.g., Design of Drugs, Bundgaard, a.ed., Elseview,1985 and Method in Enzymology, Widder, k, et al, ed.; academy, 1985, vol.42, p.309-396; bundgaard, H. "Design and Application of precursors" in ATextwood of Drug Design and Development, Krosgaard-Larsen and H.Bundgaard, Ed.,1991, Chapter 5, p.113-191; and Bundgaard, h., Advanced Drug Delivery Review,1992,8,1-38, which is incorporated herein by reference. Prodrugs described herein include, but are not limited to, the following groups and combinations of these groups; amine-derived prodrugs:

Hydroxy prodrugs include, but are not limited to, acyloxyalkyl esters, alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters, and ester-containing disulfides.

The term "pharmaceutically acceptable salt" as used herein includes salts that retain the biological effectiveness of the free acid and free base of the specified compound and are not biologically or otherwise undesirable. The compounds may have acidic or basic groups and may therefore react with some inorganic or organic bases and inorganic or organic acids to form pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts include those salts prepared by reacting a compound described herein with an inorganic or organic acid or an inorganic base, such salts including acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bromide, butyrate, butyne-1, 4-dioate, camphorate, camphorsulfonate, hexanoate, octanoate, chloride, chlorobenzoate, chlorocarbonate, and mixtures thereofA compound, a citrate salt, a cyclopentanepropionate salt, a caprate salt, a digluconate salt, a dihydrogen phosphate salt, a dinitrobenzoate salt, a dodecylsulfate salt, an ethanesulfonate salt, a formate salt, a fumarate salt, a glucoheptonate salt (glucoheptonate salt), a glycerophosphate salt, a glycolate salt, a hemisulfate salt, a heptanoate salt, a hexanoate salt, a hexyne-1, 6-dioate salt, a hydroxybenzoate salt, a gamma-hydroxybutyrate salt, a hydrochloride salt, a hydrobromide salt, a hydroiodide salt, a 2-hydroxyethanesulfonate salt, an iodide salt, an isobutyrate salt, a lactate salt, a maleate salt, a malonate salt, a methanesulfonate salt, a mandelate salt, a metaphosphate salt, a methanesulfonate salt, a methoxybenzoate salt, a methylbenzoate salt, a hydrogen phosphate salt, a 1-naphthalenesulfonate salt, a 2-naphthalenesulfonate salt, a nicotinate salt, an oxalate salt, a pamoate salt, a pectate salt (, Persulfates, phenylacetates, phenylpropionates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, pyrosulfates, pyrophosphates, propiolates, propionates, phthalates, phenylbutyrates, propanesulfonates, pyrophosphates, salicylates, succinates, sulfates, sulfites, succinates, suberates, sebacates, sulfonates, tartrates, thiocyanates, tosylates, undecanoates, and xylenesulfonates. Other acids, such as oxalic, while not pharmaceutically acceptable in themselves, may be used to prepare salts which are useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts (see, e.g., Berge et al, j. pharm. sci.1977,66, 1-19.). Furthermore, those compounds described herein that may contain free acid groups may be reacted with a suitable base such as the hydroxide, carbonate, bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali metal salts or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Illustrative examples of the base include sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, N ⁺(C_1-4Alkyl radical)₄And the like. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, dihydrogenated amine, and mixtures thereof,Piperazine, and the like. It is to be understood that the compounds described herein also include the quaternization of any basic nitrogen-containing groups they may contain. Water-soluble or oil-soluble or water-dispersible or oil-dispersible products can be obtained by such quaternization. See, e.g., Berge et al, supra. These salts may be prepared in situ during the final isolation and purification of the compounds of the invention or by separately reacting the purified compound in free base form with a suitable organic or inorganic acid and isolating the salt thus formed.

The term "enhance" as used herein means to increase or prolong the efficacy or duration of the intended effect. Thus, with respect to enhancing the effect of a therapeutic agent, the term "enhance" means the ability to increase or prolong in efficacy or persistence the effect of other therapeutic agents on the system. As used herein, "synergistic amount" means an amount sufficient to enhance the effect of another therapeutic agent in a desired system.

The terms "pharmaceutical combination," "administering other therapy," "administering other therapeutic agent," and the like, as used herein, mean a pharmaceutical treatment resulting from the mixing or combining of more than one active ingredient, and include fixed and non-fixed combinations of active ingredients. The term "fixed combination" means that at least one compound described herein and at least one auxiliary agent are administered to a patient simultaneously in the form of a single entity or dose. The term "non-fixed combination" means that at least one compound described herein and at least one adjuvant are administered to a patient as separate entities either simultaneously, concurrently or sequentially with variable time intervals, wherein such administration provides effective levels of the two or more compounds in the patient. These are also suitable for combination therapy, for example administration of three or more active ingredients.

The terms "co-administration," "co-administration with …," and grammatical equivalents thereof, as used herein, are intended to include the administration of a selected therapeutic agent to a single patient, and are intended to include treatment regimens in which the agents are administered by the same or different routes of administration, or at the same or different times. In some embodiments, the compounds described herein will be administered simultaneously with other agents. These terms include administering two or more agents to an animal such that the agents and/or their metabolites are present in the animal at the same time. They include simultaneous administration in separate compositions, administration at different times in separate compositions, and/or administration in a composition in which both agents are present. Thus, in some embodiments, the compounds of the present invention and other agents are administered as a single composition. In some embodiments, the compounds of the present invention and other agents are mixed in the composition.

The term "metabolite" as used herein means a derivative of a compound that is formed when the compound is metabolized.

The term "active metabolite" as used herein means a biologically active derivative of a compound that is formed when the compound is metabolized.

The term "metabolized" as used herein means the sum of processes (including, but not limited to, hydrolysis reactions and enzyme-catalyzed reactions) in which a particular substance is biologically altered. Thus, enzymes can produce specific structural changes to a compound. For example, cytochrome P450 catalyzes a variety of oxidation and reduction reactions, while uridine diphosphate glucuronosyltransferase catalyzes the transfer of an activated glucuronic acid molecule to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines and free sulfhydryl groups. Other information about metabolism is available from The Pharmacological Basis of therapeutics, 9 th edition, McGraw-Hill (1996).

Compound (I)

Described herein are compounds of formula I and pharmaceutically acceptable salts, solvates, polymorphs, esters, amides, tautomers, or prodrugs thereof:

wherein

Z is H or F;

x is F, Cl, CH₃、CH₂OH、CH₂F、CHF₂Or CF₃；

All said phenyl or Het groups being optionally substituted by F, Cl, Br, I, acetyl, methyl, CN, NO ₂、CO₂H、C₁-C₃Alkyl radical, C₁-C₃Alkoxy radical, C₁-C₃alkyl-C (= O) -, C₁-C₃alkyl-C (= S) -, C₁-C₃alkoxy-C (= S) -, C₁-C₃alkyl-C (= O) O-, C₁-C₃alkyl-O- (C = O) -, C₁-C₃alkyl-C (= O) NH-, C₁-C₃alkyl-C (= NH) NH-, C₁-C₃alkyl-NH- (C = O) -, di-C₁-C₃alkyl-N- (C = O) -, C₁-C₃alkyl-C (= O) N (C)₁-C₃Alkyl) -, C₁-C₃alkyl-S (= O)₂NH-or trifluoromethyl;

all of said methyl groups being optionally substituted with 1, 2 or 3F atoms;

R⁰the method comprises the following steps: H. f, Cl, Br, I, CH₃NH-、(CH₃)₂N-、C₁-C₆Alkyl radical, C₁-C₄Alkoxy radical, C₃-C₆Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, phenyl, monosubstituted phenyl, O (C)₁-C₄Alkyl), O-C (= O) (C)₁-C₄Alkyl) or C (= O) O (C)₁-C₄Alkyl groups); wherein

G₁Is optionally substituted by an amino group, C₁-C₃Alkylamino or dialkylamino radical substituted C₁-C₆An alkyl group, said dialkylamino group comprising 2C's which can be the same or different ₁-C₄An alkyl group; or

G₁Is C₃-C₈A diaminoalkyl group;

G₂is a saturated, unsaturated or aromatic 5-or 6-membered ring containing 1-3 ring heteroatoms independently selected from N, O and S, optionally independently selected from F, Cl, OH, O (C)₁-C₃Alkyl), OCH₃、OCH₂CH₃、CH₃C(=O)NH、CH₃C(=O)O、CN、CF₃And 1-3 substituents of a 5-membered aromatic heterocyclic group containing 1-4 ring heteroatoms independently selected from N, O and S;

R_1ais methyl, optionally substituted by 1 to 3 fluorine atoms or 1 to 3 chlorine atoms, or OH, cyclopropoxy or C₁-C₃Alkoxy substituted, wherein said cyclopropoxy group or said C₁-C₃C of alkoxy radicals₁-C₃Alkyl moiety optionally substituted with one hydroxy or methoxy group, and wherein said C₁-C₄All C in alkoxy₃-the alkyl group is optionally further substituted with another OH group;

R_1cis (CH)₂)_nO_mR'; wherein

m is 0 or 1; and wherein

When m is 0, n is 1 or 2;

when m is 1, n is 2 or 3;

R_1dis C (A), (A') (B) -; wherein

B is H or C_1-4Alkyl, optionally substituted with one or two OH groups;

A and A' form a 3-6 membered saturated ring together with the carbon atom to which they are attached;

R_1eis that

Wherein

q is 1 or 2;

R₂and R₃Each independently is: H. f, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl or methylsulfonyl;

R₄the method comprises the following steps: H. f, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, N-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl, methylsulfonyl, nitro, acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazole, 1,3, 4-thiadiazole, 5-methyl-l, 3, 4-thiadiazole, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl and N-pyrrolidinylcarbonylamino;

R₅is H, F, Cl or methyl;

R₆is H, F, Cl or methyl;

Ar₁is that

Wherein

U and V are independently N, CR₂Or CR₃；

R₂、R₃And R₄Independently are: H. f, Cl, Br, CH₃、CH₂F、CHF₂、CF₃OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl, acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl A group selected from the group consisting of methyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazolyl, 1,3, 4-thiadiazolyl, 5-methyl-l, 3, 4-thiadiazolyl, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrolidinylcarbonylamino, and methanesulfonyl;

R₅and R₆Independently H, F, Cl or methyl;

Ar₂is that

Wherein

u is-S-, -O-or-N =, and wherein

When U is-O-or-S-, V is-CH =, -CCl = or-N =;

when U is-N = V is-CH =, -CCl =, or-N =;

R₇is H or methyl;

R₈is H, acetamido, methyl, F or Cl;

Ar₃is that

Wherein

U is-NH-, -NCH₃-or-O-;

R₇and R₈Independently H, F, Cl or methyl.

Except that for the group G, R herein⁰X, Y and Z, including others that may occur to those skilled in the chemical and pharmaceutical artsAnd (4) substitution.

In some embodiments, the present invention provides compounds of formula I wherein G is G₁Or G₂. In other embodiments, G is G₁. In further or other embodiments, G is G₂。

In some embodiments, the present invention provides compounds of formula I wherein G is R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃. In further or other embodiments, G is R _1a、R_1b、R_1c、R_1dOr R_1e. In further or other embodiments, G is R_1a. In further or other embodiments, G is R_1b. In further or other embodiments, G is R_1c. In further or other embodiments, G is R_1d. In further or other embodiments, G is R_1e. In further or other embodiments, G is Ar₁、Ar₂Or Ar₃. In further or other embodiments, G is Ar₁. In further or other embodiments, G is Ar₂. In further or other embodiments, G is Ar₃。

In some embodiments, there is provided a compound of formula I or a pharmaceutically acceptable salt thereof. In further or other embodiments, provided herein are compounds of formula I or solvates thereof. In further or other embodiments, provided herein are compounds of formula I or their polymorphs. In further or other embodiments, provided herein are compounds of formula I or their esters. In further or other embodiments, provided herein are compounds of formula I or their amides. In further or other embodiments, provided herein are compounds of formula I or tautomers thereof. In further or other embodiments, provided herein are compounds of formula I or prodrugs thereof.

In some embodiments, Z is H. In some embodiments, Z is F. In some embodiments, X is F. In some embodiments, X is Cl. In some embodiments, X is CH₃. In some embodiments, X is CH₂And (5) OH. In some embodiments, X is CH₂F. In some embodiments, X is CHF₂. In some embodiments, X is CF₃. In some embodiments, X is F, Cl or CH₃。

In some embodiments, G is G₁Or G₂X is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃、C₁-C₃Alkyl, phenyl, pyridyl, pyrrolyl, pyrazolyl optionally substituted by F, Cl, Br, I, acetyl, methyl, CN, NO₂、CO₂H、C₁-C₃Alkyl radical, C₁-C₃Alkoxy radical, C₁-C₃alkyl-C (= O) -, C₁-C₃alkyl-C (= S) -, C₁-C₃alkoxy-C (= S) -, C₁-C₃alkyl-C (= O) O-, C₁-C₃alkyl-O- (C = O) -, C₁-C₃alkyl-C (= O) NH-, C₁-C₃alkyl-C (= NH) NH-, C₁-C₃alkyl-NH- (C = O) -, di-C₁-C₃alkyl-N- (C = O) -, C₁-C₃alkyl-C (= O) N (C)₁-C₃Alkyl) -, C₁-C₃alkyl-S (= O)₂NH-or trifluoromethyl; and Z is H or F. In further or other embodiments, G is G₁Or G₂And R is⁰Is F, Cl, C₁-C₄Alkyl or C₁-C₄Alkoxy radical, said C₁-C₄Alkyl radical and said C ₁-C₄C of alkoxy radicals₁-C₄Alkyl moieties optionally substituted by F, Cl, OCH₃Or OCH₂CH₃And (4) substitution. In further or other embodiments, G is G₁Or G₂And R is⁰Is H, F, Cl, C₁-C₄Alkyl radicalMethoxy, ethoxy or 2-methoxy-ethoxy.

In some embodiments, G₁Is N-methyl-2-aminoethyl. In further or other embodiments, G₁Is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1, 2 or 3. In further or other embodiments, G₁Is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1, 2 or 3 and X is F. In further or other embodiments, G₁Is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, wherein n is 1, 2 or 3, X is F and Z is F.

In some embodiments, G₂Is 1-piperidinyl, 2-piperidinyl, 3-piperidinyl or 4-piperidinyl. In further or other embodiments, G₂Is morpholinyl, 1-piperazinyl or 2-piperazinyl.

In some embodiments, G is R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃And X is F, Cl or CH₃. In further or other embodiments, G is R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃X is F, Cl or CH₃And Y is I, Br, Cl, CF₃Or C₁-C₃An alkyl group. In further or other embodiments, G is R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃X is F, Cl or CH₃Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, and Z is H or F.

In further or other embodiments, G is R _1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃And R is⁰Is F, Cl, C₁-C₄Alkyl or C₁-C₄Alkoxy radical, said C₁-C₄Alkyl radical and said C₁-C₄C of alkoxy radicals₁-C₄Alkyl moieties optionally substituted by F, Cl, OCH₃Or OCH₂CH₃And (4) substitution. In further or other embodiments, G is R_1a、R_1b、R_1c、R_1d、R_1e、Ar₁、Ar₂Or Ar₃And R is⁰Is H, F, Cl, C₁-C₄Alkyl, methoxy, ethoxy or 2-methoxy-ethoxy.

In some embodiments, G is R_1a(ii) a And Z is F. In further or other embodiments, G is R_1aWherein R is_1aIs CH₃，R⁰Is H; and Y is Br, I, CF₃Or CH₃. In some embodiments, G is R_1bAnd Z is F. In further or other embodiments, G is R_1bZ is F, and R⁰Is H, F or OCH₃. In further or other embodiments, G is R_1bZ is F, R⁰Is H, F or OCH₃And X is F or CH₃. In further or other embodiments, G is R_1bZ is F, R⁰Is H, F or OCH₃X is F or CH₃And Y is Br, I or CH₃. In further or other embodiments, G is R_1bWherein R is_1bIs C₃-C₆A cycloalkyl group. In further or other embodiments, G is R_1bWherein R is_1bIs substituted C₃-C₆A cycloalkyl group. In further or other embodiments, G is R_1bWherein R is_1bIs unsubstituted C₃-C₆A cycloalkyl group. In further or other embodiments, G is R _1bWherein R is_1bIs unsubstituted C₃-C₆Cycloalkyl radical and R⁰Is H. In further or other embodiments, G is R_1bWherein R is_1bIs isopropyl or cyclopropyl.

In some embodiments, G is R_1cAnd Y is I, Br, CH₃Or CF₃. In further or other embodiments, G is R_1cY is I, Br, CH₃Or CF₃And Z is F. In further or other embodiments, G is R_1cY is I, Br, CH₃Or CF₃Z is F and m is 0.

In some embodiments, G is R_1dAnd R is⁰Is fluorine, chlorine, methyl, ethyl, propyl, isopropyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino. In further or other embodiments, G is R_1d，R⁰Is fluorine, chlorine, methyl, ethyl, propyl, isopropyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino, and X is F, Cl, CH₃Or a monofluoromethyl, difluoromethyl or trifluoromethyl group. In further or other embodiments, G is R_1d，R⁰Is fluorine, chlorine, methyl, ethyl, propyl, isopropyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino, X is F, Cl, CH ₃Or monofluoromethyl, difluoromethyl or trifluoromethyl and Y is I, Br, Cl or monofluoromethyl, difluoromethyl or trifluoromethyl. In further or other embodiments, G is R_1d，R⁰Is fluorine, chlorine, methyl, ethyl, propyl, isopropyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino, X is F, Cl, CH₃Or monofluoromethyl, difluoromethyl or trifluoromethyl, Y is I, Br, Cl or monofluoromethyl, difluoromethyl or trifluoromethyl and Z is H or F. In further or other embodiments, G is R_1dAnd R is⁰Is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxyA group-ethoxy group.

In further or other embodiments, G is R_1d，R⁰Is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy, and X is F, Cl or CH₃. In further or other embodiments, G is R_1d，R⁰Is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy, X is F, Cl or CH₃And Y is I, Br, Cl or monofluoromethyl, difluoromethyl or trifluoromethyl. In further or other embodiments, G is R _1d，R⁰Is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy, X is F, Cl or CH₃Y is I, Br, Cl or monofluoromethyl, difluoromethyl or trifluoromethyl and Z is H or F. In further or other embodiments, G is R_1dAnd R is⁰Is H; x is F, Cl, CH₃Or a monofluoromethyl, difluoromethyl or trifluoromethyl group. In further or other embodiments, G is R_1d，R⁰Is H; x is F, Cl, CH₃Or monofluoromethyl, difluoromethyl or trifluoromethyl and Y is I, Br, Cl or monofluoromethyl, difluoromethyl or trifluoromethyl. In further or other embodiments, G is R_1d，R⁰Is H; x is F, Cl, CH₃Or monofluoromethyl, difluoromethyl or trifluoromethyl, Y is I, Br, Cl or monofluoromethyl, difluoromethyl or trifluoromethyl and Z is H or F.

In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆A group of cycloalkyl groups. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is the radical of H. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is methyl, ethyl, 2-hydroxyethyl, n-propyl, 3-hydroxypropyl, 2, 3-dihydroxypropyl, 3, 4-dihydroxybutyl, isopropyl, l-methyl-2-hydroxy Ethyl, n-butyl, sec-butyl, isobutyl or 2-hydroxymethyl-3-hydroxypropyl groups.

In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is the radical of 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is the radical of 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, where the chiral carbon in B is in the R configuration. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is the radical of 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, where the chiral carbon in B is in the S configuration. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is methyl optionally substituted by one OH group, or C optionally substituted by one or two OH groups₂-C₄The radical of an alkyl group. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Radical of cycloalkyl and R⁰Is fluorine, chlorine, methyl, ethyl, propyl, isopropyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino. In further or other embodiments, G is R _1dWherein R is_1dIs C (A), (A') is C₁-C₆Radical of cycloalkyl and R⁰Is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy. In further or other embodiments, G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Radical of cycloalkyl and R⁰Is H; x is F, Cl, CH₃Or a monofluoromethyl, difluoromethyl or trifluoromethyl group.

In a further or alternative embodiment, the invention provides a composition comprising a compound of formula IComposition wherein G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is a radical of 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the R configuration, said composition being substantially free of the S isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is a 2, 3-dihydroxypropyl group, wherein the chiral carbon in B is in the R configuration, said composition being substantially free of the S isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is the radical of a 3, 4-dihydroxybutyl wherein the chiral carbon in B is in the R configuration, said composition being substantially free of the S isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R _1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is a radical of 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the S configuration, said composition being substantially free of the R isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is a 2, 3-dihydroxypropyl group, wherein the chiral carbon in B is in the S configuration, said composition being substantially free of the R isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs C (A), (A') is C₁-C₆Cycloalkyl and B is the radical of a 3, 4-dihydroxybutyl in which the chiral carbon in B is in the S configuration, said composition being substantially free of the R isomer.

In further or other embodiments, G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl. In further or other embodiments, G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and B is H. In further or other embodiments, G is R_1dWherein R is_1dIs a radical in which C (A), (A') is cyclopropyl and B is methyl, ethyl, 2-hydroxyethyl, n-propyl, 3-hydroxypropyl, 2, 3-dihydroxypropyl, 3, 4-dihydroxybutyl, isopropyl, l-methyl-2-hydroxyethyl, n-butyl, sec-butyl, isobutyl or 2-hydroxymethyl-3-hydroxypropyl. In further or other embodiments, G is R _1dWherein R is_1dIs a radical in which C (A), (A') is cyclopropyl and B is 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl. In further or other embodiments, G is R_1dWherein R is_1dIs a radical in which C (A), (A') is cyclopropyl and B is 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the R configuration. In further or other embodiments, G is R_1dWherein R is_1dIs a radical in which C (A), (A') is cyclopropyl and B is 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the S configuration. In further or other embodiments, G is R_1dWherein R is_1dIs C (A) (A') is cyclopropyl and B is methyl optionally substituted with one OH group, or C optionally substituted with one or two OH groups₂-C₄The radical of an alkyl group. In further or other embodiments, G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and R⁰Is a radical of fluorine, chlorine, methyl, ethyl, propyl, isopropyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino. In further or other embodiments, G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and R ⁰Is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy. In further or other embodiments, G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and R⁰Is H; x is F, Cl, CH₃Or a monofluoromethyl, difluoromethyl or trifluoromethyl group.

In advance ofIn one or other embodiments, the present invention provides compositions comprising compounds of formula I wherein G is R_1dWherein R is_1dIs a group wherein C (A) (A') is cyclopropyl and B is 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the R configuration, said composition being substantially free of the S isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and B is 2, 3-dihydroxypropyl, wherein the chiral carbon in B is in the R configuration, said composition being substantially free of the S isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and B is 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the R configuration, said composition being substantially free of the S isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R _1dWherein R is_1dIs a group wherein C (A) (A') is cyclopropyl and B is 2, 3-dihydroxypropyl or 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the S configuration, said composition being substantially free of the R isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs a group wherein C (A) (A') is cyclopropyl and B is 2, 3-dihydroxypropyl, wherein the chiral carbon in B is in the S configuration, said composition being substantially free of the R isomer. In a further or other embodiment, the present invention provides a composition comprising a compound of formula I, wherein G is R_1dWherein R is_1dIs a group wherein C (A), (A') is cyclopropyl and B is 3, 4-dihydroxybutyl, wherein the chiral carbon in B is in the S configuration, said composition being substantially free of the R isomer.

In some embodiments, G is R_1eAnd n is 1. In further or other embodiments, G is R_1e，R⁰Is H, R_4-6Is H, R₂And R₃Independently H, F, Cl, Br, CH₃、CH₂F、CHF₂、CF₃、OCH₃、OCH₂F、OCHF₂、OCF₃Ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, sec-butyl, tert-butyl and methanesulfonyl, X is F and Y is I.

In some embodiments, G is Ar₁Wherein Ar is₁Is phenyl optionally substituted with one group selected from acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazolyl, 1,3, 4-thiadiazolyl, 5-methyl-1, 3, 4-thiadiazolyl, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrolidinylcarbonylamino and methanesulfonyl, optionally substituted with one group independently selected from F, Cl and CH ₃1-3 substituents of (A). In further or other embodiments, G is Ar₁Wherein Ar is₁Is phenyl optionally substituted with one group selected from acetylamino, amidino, cyano, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, l,3, 4-oxadiazol-2-yl, 5-methyl-l, 3, 4-oxadiazolyl, 1,3, 4-thiadiazolyl, 5-methyl-1, 3, 4-thiadiazolyl, lH-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrolidinylcarbonylamino and methanesulfonyl, optionally substituted with one group independently selected from F, Cl and CH₃Is substituted by 1 to 3 substituents of (A), R⁰Is H, X is F, Cl or methyl and Y is Br, I, CF₃、C₁-C₃Alkyl radical, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, cyclopropyl, OCH₃、OCH₂CH₃Or SCH₃. In some embodiments, G is Ar₁Wherein Ar is₁Is thatAnd wherein R₂And R₃Independently H, F, Cl, CH₃、CF₃、OCH₃. In further or other embodiments, G is Ar₁Wherein Ar is₁Is thatAnd wherein R₂And R₃Independently H, F, Cl, CH₃、CF₃、OCH₃X is F or CH₃Y is I, Br or Cl; and Z is F. In further or other embodiments, G is Ar₁Wherein Ar is₁Is phenyl or monosubstituted phenyl. In further or other embodiments, G is Ar₁Wherein Ar is ₁Is phenyl or monosubstituted phenyl, X is F or CH₃Y is I, Br or Cl, Z is F; and R is⁰Is F, methyl, ethyl, methoxy or 2-methoxy-ethoxy. In further or other embodiments, G is Ar₁Wherein U is N or CR₂And V is N. In further or other embodiments, G is Ar₁Wherein U is N or CR₂And V is CR. In further or other embodiments, G is Ar₁Wherein U is N or CR₂V is CR, R⁰Is H, X is F, Cl or methyl and Y is Br, I, CF₃、C₁-C₃Alkyl radical, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, cyclopropyl, OCH₃、OCH₂CH₃Or SCH₃。

In some embodiments, G is Ar₂Wherein Ar is₂Is thatWherein R is₇Is H or methyl and R₈Is H, acetamido, methyl, F or Cl. In further or other embodiments, G is Ar₂Wherein Ar is₂Is thatWherein R is₇Is H or methyl, R₈Is H, acetylamino, methyl, F or Cl, R⁰Is H, X is F, Cl or methyl, Y is Br, I, CF₃、C₁-C₃Alkyl radical, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, cyclopropyl, OCH₃、OCH₂CH₃Or SCH₃And Z is F. In further or other embodiments, G is Ar₂Wherein Ar is₂Is thatWherein U is S or O, V is CH =, and R₈Is H or CH₃，R₇Is H or methyl, R₈Is H, acetylamino, methyl, F or Cl, R⁰Is H, X is F, Cl or methyl, Y is Br, I, CF ₃、C₁-C₃Alkyl radical, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, cyclopropyl, OCH₃、OCH₂CH₃Or SCH₃And Z is F. In further or other embodiments, R⁰Is H. In further or other embodiments, R⁰Is H, X is F or Cl and Y is Br, I, CH₂CH₃Or SCH₃。

In some embodiments, G is Ar₃Wherein U is-O-.

In further or other embodiments, G is R_1aWherein R is_1aAs defined above. In further or other embodiments, G is R_1aAnd R is⁰Is H, wherein R_1aAs defined above. In further or other embodiments, G is R_1aAnd R is⁰Is as defined above except for H, and R_1aAs defined above. In further or other embodiments, G is R_1aWherein R is_1aIs methyl, monohalomethyl, C₁-C₃Alkoxymethyl or cyclopropoxymethyl. In further or other embodiments, G is R_1aWherein R is_1aIs methyl, monohalomethyl, C₁-C₃Alkoxymethyl or cyclopropoxymethyl and wherein R⁰Is F, Cl, C₁-C₃Alkyl, monochloro C₁-C₃Alkyl radical, C₁-C₃Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy.

In further or other embodiments, G is R_1bWherein R is_1bAs defined above. In further or other embodiments, G is R_1bAnd R is⁰Is H, wherein R _1bAs defined above. In further or other embodiments, G is R_1b，R⁰Is H and Z is F, wherein R_1bAs defined above. In further or other embodiments, G is R_1bAnd R is⁰Is as defined above except for H, and R_1bAs defined above. In further or other embodiments, G is R_1bWherein R is_1bIs isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, all optionally independently selected from F, Cl, OH and OCH₃Substituted with 1 or 2 substituents of (a); y is Br, I, methyl or trifluoromethyl. In further or other embodiments, G is R_1bWherein R is_1bIs isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, optionally independently selected from F, Cl, OH and OCH₃Substituted with 1 or 2 substituents of (a); y is Br, I, methyl or trifluoromethyl; and R is⁰The method comprises the following steps: F. cl, C₁-C₃Alkyl, monochloro C₁-C₃Alkyl radical, C₁-C₃Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In further or other embodiments, G is R_1bWherein R is_1bIs isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, all optionally substituted by 1 Cl or by 1 or 2 OH groups; and Y is Br, I, methyl or trifluoromethyl. In further or other embodiments, G is R _1bWherein R is_1bIs isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, all optionally substituted by 1 Cl or by 1 or 2 OH groups; y is Br, I, methyl or trifluoromethyl; and R is⁰Is F, Cl, C₁-C₃Alkyl, monochloro C₁-C₃Alkyl radical, C₁-C₃Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy.

In further or other embodiments, G is R_1cWherein R is_1cAs defined above. In further or other embodiments, G is R_1cAnd R is⁰Is H, wherein R_1cAs defined above. In further or other embodiments, G is R_1cAnd R is⁰Is as defined above except for H, and R_1cAs defined above. In further or other embodiments, G is R_1cAnd R is⁰Is H, wherein R_1cIs (CH)₂)_nO_mR 'wherein m is 0 or 1, n is 2 or 3 when m is 1, n is 1 or 2 when m is 0, and R' is C₁-C₆Alkyl, optionally independently selected from F, Cl, OH, OCH₃、OCH₂CH₃And C₃-C₆1-3 substituents of the cycloalkyl group. In another more specific subgeneric embodiment, m is 0, n is 1 or 2, and R' is C₁-C₄Alkyl, which is optionally substituted as described above. In another more specific subgeneric embodiment, m is 1, n is 2 or 3, and R' is C ₁-C₄Alkyl, which is optionally substituted as described above. In a still more specific subgeneric embodiment, m is 0, n is 1 or 2, and R' is C₁-C₄Alkyl, optionally substituted by one or more substituents selected from OH, OCH₃Cl and cyclopropyl.

In further or other embodiments, G is R_1dWherein R is_1dAs defined above. In further or other embodiments, G is R_1dAnd R is⁰Is H, wherein R_1dAs defined above. In further or other embodiments, G is R_1dAnd R is⁰Is as defined above except for H, and R_1dAs defined above. In further or other embodiments, G is R_1dAnd R is⁰Is H, wherein R_1dIs C (A) (A ') (B) -, wherein B, A and A' are independently H or C optionally substituted with one or two OH groups or halogen atoms_1-4Alkyl, or A and A' together with the carbon atom to which they are attached form a 3-6 membered saturated ring, optionally containing one or two heteroatoms independently selected from O, N and S, and optionallyIndependently substituted with one or two groups independently selected from methyl, ethyl, fluoro, chloro, bromo and iodo.

In further or other embodiments, G is R_1eWherein R is_1eAs defined above. In further or other embodiments, G is R _1eAnd R is⁰Is H, wherein R_1eAs defined above. In further or other embodiments, G is R_1eAnd R is⁰Is as defined above except for H, and R_1eAs defined above.

In further or other embodiments, G is Ar₁Wherein Ar is₁As defined above. In further or other embodiments, G is Ar₁And R is⁰Is H, wherein Ar₁As defined above. In further or other embodiments, G is Ar₁And R is⁰Is as defined above except for H, and Ar₁As defined above.

In further or other embodiments, G is Ar₂Wherein Ar is₂As defined above. In further or other embodiments, G is Ar₂And R is⁰Is H, wherein Ar₂As defined above. In further or other embodiments, G is Ar₂And R is⁰Is as defined above except for H, and Ar₂As defined above.

In further or other embodiments, X is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, and Z is H or F. In further or other embodiments, X is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, Z is H or F, and R⁰Is halogen, C₁-C₆Alkyl, monohalo C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, phenyl, monosubstituted phenyl, OR₃、O-C(=O)R₄OR C (= O) OR₅. In a further or alternative embodiment, the first and second electrodes are, X is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, Z is H or F, and R⁰Is furyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl. In further or other embodiments, X is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, Z is H or F, and R⁰Is F, Cl, C₁-C₄Alkyl radical, C₁-C₃Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy.

In another more specific subgeneric embodiment, R_1dIs cycloalkyl or 1-alkyl-cycloalkyl, wherein the 1-alkyl group is optionally substituted by one or two OH groups or by one or two halogen atoms.

In another more specific subgeneric embodiment, R⁰Is halogen, C₁-C₆Alkyl, monohalo C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, phenyl, monosubstituted phenyl, OR₃、O-C(=O)R₄OR C (= O) OR₅(ii) a And R is_1dIs cycloalkyl or 1-alkyl-cycloalkyl, wherein the 1-alkyl group is optionally substituted by one or two OH groups or by one or two halogen atoms.

In another more specific subgeneric embodiment, R⁰Is furyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl; and R is _1dIs cycloalkyl or 1-alkyl-cycloalkyl, wherein the 1-alkyl group is optionally substituted by one or two OH groups or by one or two halogen atoms.

In another more specific subgeneric embodiment, R_1dIs cycloalkyl or 1-alkyl-cycloalkyl, wherein the 1-alkyl group is optionally substituted with one or two OH groups, and wherein Y is Br, I, methyl or trifluoromethyl. In another more specific subgenusIn embodiments, R_1dIs cycloalkyl or 1-alkyl-cycloalkyl, wherein the 1-alkyl group is optionally substituted with one or two fluorine or chlorine atoms, and wherein Y is Br, I, methyl or trifluoromethyl. In another more specific subgeneric embodiment, R_1dIs cycloalkyl or (1-alkyl)) -cycloalkyl, wherein said 1-alkyl group is optionally substituted with one or two OH groups, and wherein R is⁰' is: F. cl, C₁-C₃Alkyl, monochloro C₁-C₃Alkyl radical, C₁-C₃Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In another more specific subgeneric embodiment, R_1dIs tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl, each optionally substituted as described above, and wherein Y is Br, I, methyl or trifluoromethyl. In another more specific subgeneric embodiment, R _1dIs oxazolidinyl, thiazolidinyl, isoxazolidinyl, isothiazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl, each optionally substituted as described above, and wherein Y is Br, I, methyl or trifluoromethyl. In another more specific subgeneric embodiment, R_1dIs cyclopropyl or l-alkyl-cyclopropyl, wherein said 1-alkyl group is optionally substituted with one or two OH groups, and wherein R is⁰' is F, Cl, methyl, ethyl, chloromethyl, C₁-C₂Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In an even more particular embodiment, R_1dIs a l- (monohydroxyalkyl) cycloalkyl group. In another more specific embodiment, R_1dIs l- (monohydroxyalkyl) cycloalkyl, wherein R⁰' is F, Cl, methyl, ethyl, chloromethyl, C₁-C₂Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In an even more particular embodiment, R_1dIs l- (dihydroxyalkyl) cycloalkyl. In another more specific embodiment, R_1dIs l- (dihydroxyalkyl) cycloalkyl, wherein R⁰' is F, Cl, methyl, ethyl, chloromethyl, C₁-C₂Alkoxy, trifluoromethylOxy or 2-methoxy-ethoxy.

In a more specific subgeneric embodiment, U is CR₂And V is N. In another more specific subgeneric embodiment, both U and V are N. In a more specific subgeneric embodiment, U is CR₂And V is CR₃。

In a still more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar₁And Ar₁Is phenyl or monosubstituted phenyl, R⁰Is F, methyl, ethyl, C₁-C₃Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy; x is F, Cl or CH₃(ii) a Y is I; and Z is F. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar₁Wherein Ar is₁Is phenyl or monosubstituted phenyl, R⁰Is halogen, C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, all such alkyl, cycloalkyl, alkenyl and alkynyl groups optionally substituted with 1-3 substituents independently selected from halo, OH, CN, cyanomethyl, nitro, phenyl and trifluoromethyl; or R⁰Is phenyl, OR₃Furyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl. In a more specific subgeneric embodiment, the invention provides a compound of formula I wherein A is Ar ₁Wherein Ar is₁Is phenyl or monosubstituted phenyl, R⁰Is F, Cl, C₁-C₃Alkyl radical, C₁-C₃Alkoxy, 2-methoxyethoxy, C₂-C₃Alkenyl radical, C₂-C₃Alkynyl, trifluoromethyl, phenyl, furyl or thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl; x is F, Cl or methyl; y is I, Br, Cl, CF₃Or C₁-C₃An alkyl group; and Z is F.

In another still more specific subgeneric embodiment, the invention providesA compound of formula I wherein G is Ar₁Wherein Ar is₁Is phenyl or monosubstituted phenyl, R⁰Is H; x is F, Cl or CH₃(ii) a Y is Br or I; and Z is F.

In another subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar₂Wherein Ar is₂Is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylcarbamoyl, acetylamino, acetyl, methyl, ethyl, trifluoromethyl or halogen. In a more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar₂Wherein Ar is₂Is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylcarbamoyl, acetylamino, acetyl, methyl, ethyl, trifluoromethyl or halogen; r ⁰Is not H; x is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, and Z is H or F. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar₂Wherein Ar is₂Is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylcarbamoyl, acetylamino, acetyl, methyl, ethyl, trifluoromethyl or halogen; r⁰Is F, Cl, C₁-C₃Alkyl, monochloro C₁-C₃Alkyl radical, C₁-C₃Alkoxy, trifluoromethoxy, methoxy-methoxy or 2-methoxy-ethoxy; x is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, and Z is H or F. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar₂Wherein Ar is₂Is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylcarbamoyl, acetylamino, acetyl, methyl, ethyl, trifluoromethyl or halogen; r⁰Is H; x is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, and Z is H or F. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is Ar ₂Wherein Ar is₂Is thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl, all optionally substituted with methoxycarbonyl, methylcarbamoyl, acetylamino, acetyl, methyl, ethyl, trifluoromethyl or halogen; r⁰Is H or methoxy; x is F, Cl or CH₃(ii) a Y is I, Br, Cl, CF₃Or C₁-C₃Alkyl, and Z is H or F.

In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from

In some embodiments, the present invention provides a compound of formula I, or a pharmaceutically acceptable salt thereof, selected from:wherein the 2-OH carbon is in the R configuration.

In some embodiments, the present invention provides a compound of formula I, or a pharmaceutically acceptable salt thereof, selected from:wherein the 2-OH carbon is in the S configuration.

In a further or additional embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is

In some embodiments, the present invention provides a compound of formula I, wherein Y is phenyl, pyridinyl, or pyrazolyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein Y is substituted phenyl, pyridyl, or pyrazolyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein Y is Br or I. In a subgeneric embodiment, the invention provides a compound of formula I, wherein G is 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, or 4-piperidinyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is 1-piperazinyl or 2-piperazinyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is morpholinyl. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-2-aminoethyl. In a subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-3-amino-N-propyl. In another subgeneric embodimentThe present invention provides compounds of formula I wherein G is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1, 2 or 3. In another subgeneric embodiment, the invention provides a compound of formula I wherein G is (CH) ₃CH₂)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1 or 2. In a more specific subgeneric embodiment, the invention provides a compound of formula I, wherein G is 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, or 4-piperidinyl; r⁰Is H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is 1-piperazinyl or 2-piperazinyl; r⁰Is H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is morpholinyl; r⁰Is H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-2-aminoethyl; r⁰Is H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is N-methyl-3-amino-N-propyl; r⁰Is H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is (CH)₃)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1, 2 or 3; r⁰Is H, halogen or methoxy; x is F; and Y is I. In another more specific subgeneric embodiment, the invention provides a compound of formula I wherein G is (CH) ₃CH₂)₂N-CH₂CH₂-NH-(CH₂)_n-, where n is 1 or 2; r⁰Is H, halogen or methoxy; x is F; and Y is I.

In some embodiments, the present invention provides pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound selected from the group consisting of:or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier. In some embodiments, the compound is in the R configuration. In some embodiments, the compound is in the R configuration, which is substantially free of the S-isomer. In some embodiments, the compound is in the S configuration.

In some embodiments, the compound is in the S configuration, substantially free of the R-isomer. In some embodiments, the compound is:in some embodiments, the compound is: In some embodiments, the compound is:in some embodiments, the compound is:

list of non-limiting examples of compounds of formula I

The following table shows examples of the individual compounds provided or referred to in the present invention. These examples should in no way be construed as limiting examples.

Table 1 shows embodiments of the compounds of formula I according to the invention, wherein R⁰G is R, as defined herein_1aWherein R is_1aAs defined in the table, and X, Y and Z are defined in the table.

TABLE 1

Table 2 shows embodiments of the compounds of formula I according to the invention, wherein R⁰G is R, as defined herein_1bWherein R is_1bAs defined in the table, and X, Y and Z are defined in the table.

TABLE 2

Table 3 shows embodiments of the compounds of formula I according to the invention, wherein R⁰G is R, as defined herein_1cWherein R is_1cAs defined in the table, and X, Y and Z are defined in the table.

TABLE 3

Tables 4a and 4b show embodiments of compounds of formula I according to the invention, wherein G = R_1dZ is F, X is F, and R is defined in the table_1dAnd R⁰. Each row in the table corresponds to five substances that differ only in the Y position.

TABLE 4a

TABLE 4b

CMPD#	A,A′	B	R⁰
				1(a-d)	H,H	H	2-furyl radical
2(a-d)	H,H	H	1,2, 3-triazol-4-yl
				3(a-d)	H,H	H	4-imidazolyl
4(a-d)	H,H	H	2-furyl radical
				5(a-d)	H,H	H	1,2, 3-triazol-4-yl
6(a-d)	H,H	H	4-imidazolyl
				7(a-d)	H,H	-(CH₂)₂CH(OH)CH₂OH	2-furyl radical
8(a-d)	H,H	-(CH₂)₂CH(OH)CH₂OH	1,2, 3-triazol-4-yl
				9(a-d)	H,H	-(CH₂)₂CH(OH)CH₂OH	4-imidazolyl
10(a-d)	-(CH₂)₂-	-CH₂(C₃H₅)	2-furyl radical
				11(a-d)	-(CH₂)₂-	-CH₂(C₃H₅)	1,2, 3-triazol-4-yl
12(a-d)	-(CH₂)₂-	-CH₂(C₃H₅)	4-imidazolyl
				13(a-d)	-(CH₂)₂-	CH₃	4-thiazolyl group
14(a-d)	-(CH₂)₂-	-CH₂CH₂OH	4-thiazolyl group
				15(a-d)	-(CH₂)₂-	-(CH₂)₂CH(OH)CH₂OH	4-thiazolyl group
16(a-d)	CH₃,H	-(CH₂)₂CH(OH)CH₂OH	4-thiazolyl group
				17(a-d)	-(CH₂)₂-	CH₃	2-oxazolyl radical
18(a-d)	-(CH₂)₂-	-CH₂CH₂OH	2-oxazolyl radical
				19(a-d)	-(CH₂)₂-	-(CH₂)₂CH(OH)CH₂OH	2-oxazolyl radical

CMPD#	A,A′	B	R⁰
				20(a-d)	CH₃,H	-(CH₂)₂CH(OH)CH₂OH	2-oxazolyl radical
21(a-d)	H,H	H	2-furyl radical
				22(a-d)	H,H	H	1,2, 3-triazol-4-yl
23(a-d)	H,H	H	4-imidazolyl
				24(a-d)	H,H	H	2-furyl radical
25(a-d)	H,H	H	1,2, 3-triazol-4-yl
				26(a-d)	H,H	H	4-imidazolyl
27(a-d)	H,H	-CH₂CH(OH)CH₂OH	2-furyl radical
				28(a-d)	H,H	-CH₂CH(OH)CH₂OH	1,2, 3-triazol-4-yl
29(a-d)	H,H	-CH₂CH(OH)CH₂OH	4-imidazolyl
				30(a-d)	-(CH₂)₂-	-CH₂(C₃H₅)	2-furyl radical
31(a-d)	-(CH₂)₂-	-CH₂(C₃H₅)	1,2, 3-triazol-4-yl
				32(a-d)	-(CH₂)₂-	-CH₂(C₃H₅)	4-imidazolyl
33(a-d)	-(CH₂)₂-	CH₃	4-thiazolyl group
				34(a-d)	-(CH₂)₂-	-CH₂CH₂OH	4-thiazolyl group
35(a-d)	-(CH₂)₂-	-(CH₂)₂CH(OH)CH₂OH	4-thiazolyl group
				36(a-d)	CH₃,H	-(CH₂)₂CH(OH)CH₂OH	4-thiazolyl group
37(a-d)	-(CH₂)₂-	CH₃	2-oxazolyl radical
				38(a-d)	-(CH₂)₂-	-CH₂CH₂OH	2-oxazolyl radical
39(a-d)	-(CH₂)₂-	-(CH₂)₂CH(OH)CH₂OH	2-oxazolyl radical
				40(a-d)	CH₃,H	-(CH₂)₂CH(OH)CH₂OH	2-oxazolyl radical

Table 5a shows embodiments of the compounds of formula I according to the invention, wherein G is Ar₁、Ar₂Or R_1dAnd R is⁰Is H, Z is F, and G and X are defined in the table. Each row in the table corresponds to five substances that differ only at the Y position (Y)_a、Y_b、Y_c、Y_dAnd Y_e) Wherein Y is_a=SCH₃；Y_b=Br；Y_c=I；Y_d=Cl；Y_e=CH₃。

TABLE 5a

Table 5b shows embodiments of compounds of formula I according to the invention, wherein G is Ar₁、Ar₂Or R_1dAnd R is⁰Is H, Z is F, and G and X are defined in the table. Each row in the table corresponds to five substances that differ only at the Y position (Y)_a、Y_b、Y_c、Y_dAnd Y_e) Wherein Y is_a= phenyl group; y is_b= 3-substituted phenyl; y is_c= 3-pyridyl; y is_d= 4-pyridyl; y is_e= 3-pyrazolyl group.

TABLE 5b

Synthesis procedure

In another aspect, methods of synthesizing the compounds described herein are provided. In some embodiments, the compounds described herein can be prepared by the following methods. The following steps and examples are intended to illustrate those methods. Neither the steps nor the examples should be construed in any way as limiting the invention. The compounds described herein can also be synthesized using standard synthetic methods known to those skilled in the art, or using methods known in the art in combination with the methods described herein. In addition, the solvents, temperatures, and other reaction conditions described herein may be varied according to the practice and knowledge of those skilled in the art.

The starting materials for the synthesis of the compounds described herein are available from commercial sources, such as Aldrich Chemical Co, (Milwaukee, Wis.), Sigma Chemical Co, (st. Using methods and materials known to those skilled in the art, e.g. in March, A_DVANCEDO_RGANICC_HEMISTRY4 th edition (Wiley 1992); carey and Sundberg, A_DVANCEDO_RGANICC_HEMISTRY4 th edition, volumes A and B (Plenum 2000,2001) and Green and Wuts, P_ROTECTIVEG_ROUPSIN O_RGANICS_YNTHESISThe compounds described herein, as well as other related compounds having different substituents, can be synthesized as described in 3 rd edition (Wiley 1999), which is incorporated by reference in its entirety. General methods for preparing the compounds disclosed herein may be derived from reactions known in the art, and the reactions may be modified by the use of appropriate reagents and conditions, as known to the skilled artisan, in order to introduce the various groups found in the formulae provided herein. The following synthetic methods can be used as guidance.

Formation of covalent bonds by reaction of electrophiles with nucleophiles

The compounds described herein may be modified to form new functional groups or substituents using a variety of electrophiles or nucleophiles. The following table entitled "examples of covalent bonds and precursors thereof" lists selected examples of covalent bonds and precursor functional groups that are generated and can be used as a guide for various available combinations of electrophiles and nucleophiles. The precursor functional groups are denoted as electrophilic and nucleophilic groups.

Examples of covalent bonds and precursors thereof

Product of covalent bonds	Electrophiles	Nucleophiles
			Amides of amides	Active ester	Amines/anilines
Amides of amides	Acyl azides	Amines/anilines
			Amides of amides	Acyl halide	Amines/anilines
Esters as pesticides	Acyl halide	Alcohols/phenols
			Esters as pesticides	Acylacetonitriles	Alcohols/phenols
Amides of amides	Acylacetonitriles	Amines/anilines
			Imines	Aldehydes	Amines/anilines
Hydrazone compounds	Aldehydes or ketones	Hydrazines
			Oximes	Aldehydes or ketones	Hydroxylamines
Alkyl amines	Alkyl halides	Amines/anilines
			Esters as pesticides	Alkyl halides	Carboxylic acids
Thioether	Alkyl halides	Thiols
			Ethers	Alkyl halides	Alcohols/phenols
Thioether	Alkyl sulfonic acid esters	Thiols
			Esters as pesticides	Alkyl sulfonic acid esters	Carboxylic acids
Ethers	Alkyl sulfonic acid esters	Alcohols/phenols
			Esters as pesticides	Acid anhydrides	Alcohols/phenols
Amides of amides	Acid anhydrides	Amines/anilines
			Thiophenols	Aryl halides	Thiols
Aryl amines	Aryl halides	Amines as herbicides
			Thioether	Aziridines	Thiols

Boric acid esters	Borate esters	Glycols
			Amides of amides	Carboxylic acids	Amines/anilines
Esters as pesticides	Carboxylic acids	Alcohols
			Hydrazines	Hydrazides	Carboxylic acids
N-acyl ureas or anhydrides	Carbodiimides	Carboxylic acids
			Esters as pesticides	Diazoalkanes	Carboxylic acids
Thioether	Epoxide compound	Thiols
			Thioether	Halogenated amides	Thiols
Aminotriazines (Ammotriazines)	Halotriazines	Amines/anilines
			Triazinyl ethers	Halotriazines	Alcohols/phenols
Amidines	Imido ester	Amines/anilines
			Urea	Isocyanates	Amines/anilines
Carbamates, their preparation and their use	Isocyanates	Alcohols/phenols
			Thiourea	Isothiocyanates	Amines/anilines
Thioether	Maleimide	Thiols
			Phosphorous acid esters	Phosphoramidites	Alcohols
Silyl ethers	Silyl halides	Alcohols
			Alkyl amines	Sulfonic acid esters	Amines/anilines
Thioether	Sulfonic acid esters	Thiols
			Esters as pesticides	Sulfonic acid esters	Carboxylic acids
Ethers	Sulfonic acid esters	Alcohols
			Sulfonamides	Sulfonyl halides	Amines/anilines
Sulfonic acid esters	Sulfonyl halides	Phenols/alcohols

Use of protecting groups

In such reactions, reactive functional groups, such as hydroxyl, amino, imino, thio or carboxyl groups, may need to be protected in case these groups are needed in the final product, to avoid their unwanted participation in the reaction. Protecting groups are used to block some or all of the reactive groups and prevent such groups from participating in chemical reactions until the protecting group is removed. In some embodiments, each protecting group is removable by a different method. The protecting groups cleaved under completely different reaction conditions meet the need for differential removal. The protecting group can be removed by acid, base and hydrogenolysis. Groups such as trityl, dimethoxytrityl, acetal and tert-butyldimethylsilyl are acid labile and they can be used to protect carboxyl and hydroxyl reactive groups in the presence of amino groups protected with a Cbz group (which can be removed by hydrogenolysis) and a base labile Fmoc group. Carboxylic acid and hydroxyl reactive groups can be blocked with base labile groups such as, but not limited to, methyl, ethyl, and acetyl groups in the presence of amines blocked with acid labile groups such as t-butyl carbamate or with carbamates that are both acid and base stable but removable by hydrolysis.

The carboxylic acid and hydroxyl reactive groups may also be blocked with hydrolytically removable protecting groups such as benzyl groups, while the amine groups capable of forming hydrogen bonds with acids may be blocked with base labile groups such as Fmoc. As exemplified herein, carboxylic acid reactive groups may be protected by conversion to simple ester compounds, or they may be blocked with protecting groups removable by oxidation, such as 2, 4-dimethoxybenzyl, while coexisting amino groups may be blocked with fluoride-labile silyl carbamates.

Allyl protecting groups are used in the presence of acidic and basic protecting groups because it is stable and can be subsequently removed with metal or pi acid catalysts. For example, allyl-blocked carboxylic acids can be deprotected using a Pd-catalyzed reaction in the presence of acid-labile t-butyl carbamate or base-labile amine acetate protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate may be attached. As long as the residue is attached to the resin, the functional group is blocked and cannot react. Once released from the resin, the functional groups are available to react.

The protecting or blocking group may be selected from:

Other Protecting Groups, as well as details of techniques suitable for generating Protecting Groups and removing them, are described in Greene and Wuts, Protective Groups in Organic Synthesis, 3 rd edition, John Wiley & Sons, New York, NY,1999, and Kocienski, Protective Groups, Thieme Verlag, New York, NY,1994, which are incorporated herein by reference in their entirety.

Preparation of Compounds of formula I

The compounds of the present invention may be prepared by various methods. The following steps are intended to illustrate those methods and the examples given are intended to illustrate the scope of the invention. These methods and examples are not to be construed as limiting the invention in any way.

I. The preparation of the compound of formula VI is summarized below

Scheme I above illustrates a process for preparing sulfonamide derivatives of formula VI. The 1, 2-diamine derivatives (formula IV) can be easily prepared from the desired nitro derivatives (formula I) in two steps. The compound of formula IV can be reacted with sulfonyl chloride derivatives (formula V, see next scheme) to form the desired sulfonamides. Alternatively, the 1, 2-diamine derivative IV may be protected as an imidazolidinone (formula VII) prior to reaction with the corresponding sulfonyl chloride. Deprotection of 1, 2-diamine VIII under basic conditions affords the desired starting material (formula VI).

General scheme for the synthesis of compounds of formula V

Scheme II above represents an example of the preparation of complex sulfonyl chlorides. From IX, compound XX can be synthesized, which can be alkylated and converted to potassium salt XII. With SOCl₂Or POCl₃The salt is worked up to give the desired compound. Other more specific methods of preparing unique sulfonyl chloride derivatives are reported in the experimental section.

Scheme 3 outlines a general route to synthesize compounds of general formula XIII.

Scheme III above illustrates the preparation of sulfonamide derivatives of formula XIII. These compounds can be readily obtained, for example, by reacting compound VI with boronic acid under Suzuki conditions using a palladium catalyst.

Scheme 4 outlines a general route to the synthesis of compounds of general formula XIII.

Scheme IV above illustrates the preparation of sulfonamide derivatives of general formula XV. The vinyl sulfonamide (XIV) is reacted with an amine to form a derivative of formula XV.

Other forms of the Compound of formula I

Isomers of the compound of formula I

The compounds described herein may exist as geometric isomers. The compounds described herein may have one or more double bonds. The compounds given herein include all cis, trans (E) and cis (Z) isomers and their corresponding mixtures. In certain instances, the compounds may exist as tautomers. The compounds described herein include all possible tautomers of the formulae described herein. The compounds described herein may have one or more chiral centers, and each center may exist in either the R or S configuration. The compounds described herein include all diastereomeric, enantiomeric and epimeric forms, and their corresponding mixtures. In other embodiments of the compounds and methods provided herein, mixtures of enantiomers and/or diastereomers resulting from a single preparation step, combination, or interconversion can also be used for the applications described herein. The compounds described herein may be prepared as their respective stereoisomers by reacting a racemic mixture of the compounds with an optically active resolving agent to form a pair of diastereomeric compounds, separating the resulting diastereomers and recovering the optically pure enantiomers. Enantiomeric resolution can be performed using covalent diastereomeric derivatives of the compounds described herein, or alternatively dissociable complexes (e.g., crystalline diastereomeric salts) can be used. Diastereomers have different physical properties (e.g., melting points, boiling points, solubilities, reactivities, etc.) and can be readily separated by taking advantage of these differences. Diastereomers can be separated by chiral chromatography or separation/resolution techniques based on differences in solubility. The optically pure enantiomer and resolving agent are then recovered by any feasible method that does not cause racemization. A more detailed description of a method suitable for resolving stereoisomers of compounds from their racemic mixtures can be found in Jean Jacques, Andre Collet, Samuel h.wilen, "eneromers, racemes and solutions," John Wiley and Sons, inc.,1981, which is incorporated herein by reference in its entirety.

Labelled Compounds of formula I

Isotopically labeled compounds of formula I and methods of treating disorders are also described herein. For example, the invention provides methods of treating diseases by administering isotopically labeled compounds of formula I. The isotopically labeled compound of formula I can be administered in a pharmaceutical composition. Thus, compounds of formula I also include isotopically-labeled compounds, which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be included in compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as²H、³H、¹³C、¹⁴C、^l5N、¹⁸O、¹⁷O、³¹P、³²P、³⁵S、¹⁸F and³⁶C1. compounds described herein, or pharmaceutically acceptable salts thereof, that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labelled compounds of formula I, e.g. radioactive isotopes such as³H and¹⁴those compounds in which C is contained are useful in drug and/or substrate tissue distribution assays. Tritium is³H and carbon-14 i.e¹⁴The C isotope is generally easy to prepare and detect. In addition, with heavier isotopes such as deuterium ²H substitution may result in certain therapeutic advantages resulting from better metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and thusThis may be desirable in some cases. Isotopically labeled compounds and pharmaceutically acceptable salts thereof can generally be prepared by employing the procedures described herein by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.

The compounds described herein may be labeled by other methods including, but not limited to, the use of chromophores or fluorophores, bioluminescent labels, or chemiluminescent labels.

Pharmaceutically acceptable salts of the compounds of formula I

Also described herein are pharmaceutically acceptable salts of the compounds of formula I and methods of treating disorders. For example, the invention provides methods of treating diseases by administering a pharmaceutically acceptable salt of a compound of formula I. Pharmaceutically acceptable salts of the compounds of formula I may be administered in the form of pharmaceutical compositions.

Thus, the compounds described herein may be prepared as pharmaceutically acceptable salts when the acidic proton present in the parent compound is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion; or when coordinated to an organic base, form the pharmaceutically acceptable salts. Base addition salts may also be prepared by reacting the free acid form of the compounds described herein with pharmaceutically acceptable inorganic or organic bases, including, but not limited to, organic bases such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like, and inorganic bases such as aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like. In addition, salt forms of the compounds disclosed herein can be prepared using salts of the starting materials or intermediates.

In addition, the compounds described herein may be prepared as pharmaceutically acceptable salts, which are formed by reacting the free base form of the compound with pharmaceutically acceptable inorganic or organic acids, including but not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, and the like; and organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, Q-toluenesulfonic acid, tartaric acid, trifluoroacetic acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, arylsulfonic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo- [2.2.2] oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4' -methylenebis- (3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, pivalic acid, tert-butylacetic acid, dodecylsulfuric acid, dimethylolpropionic acid, malic acid, dimethylolpropionic acid, maleic acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid.

Solvates of the compounds of formula I

Solvates of the compounds of formula I and methods of treating diseases are also described. For example, the present invention provides methods of treating diseases by administering solvates of the compounds of formula I. Solvates of the compounds of formula I may be administered in the form of pharmaceutical compositions.

Solvates comprise stoichiometric or non-stoichiometric amounts of solvent and may be formed during crystallization from pharmaceutically acceptable solvents such as water, ethanol and the like. When the solvent is water, hydrates are formed, or when the solvent is an alcohol, alcoholates are formed. In the methods as described herein, solvates of the compounds described herein may be conveniently prepared or formed. By way of example only, hydrates of the compounds described herein may be conveniently prepared by recrystallization from a mixture of water/organic solvents, including but not limited to dioxane, tetrahydrofuran or methanol. In addition, the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to unsolvated forms for the purposes of the compounds and methods provided herein.

Polymorphic forms of a compound of formula I

Polymorphs of the compounds of formula I and methods of treating disorders are also described herein. For example, the present invention provides methods of treating diseases by administering polymorphs of the compound of formula I. Polymorphs of the compound of formula I may be administered in the form of pharmaceutical compositions.

Accordingly, the compounds described herein include all their crystalline forms, referred to as polymorphs. Polymorphs include different crystal packing arrangements of the same elemental composition of a compound. Polymorphs can have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability and solubility. Various factors such as recrystallization solvent, crystallization rate and storage temperature may cause a single crystal form to dominate.

Crystalline polymorph of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

The invention also relates to N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide exhibiting a specific powder X-ray diffraction pattern:the crystalline polymorph a of (a). In some embodiments, the powder X-ray diffraction pattern comprises at least about 50% of the peaks shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least about 70% of the peaks shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least about 90% of the peaks shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5.

The present invention also relates to a crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, which exhibits a specific differential scanning calorimetry pattern. In some embodiments, the particular differential scanning calorimetry pattern is substantially the same as the differential scanning calorimetry pattern shown in fig. 6. In some embodiments, the crystalline polymorph a has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry.

The present invention also relates to polymorphs of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide prepared by a process comprising the step of crystallizing amorphous N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide from a solvent. The present invention also relates to polymorphs of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide prepared by a process comprising the step of crystallizing amorphous N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide from a mixture of hexane and ethyl acetate.

The present invention also relates to pharmaceutical compositions comprising an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and a pharmaceutically acceptable carrier or vehicle. Other aspects of the invention relate to pharmaceutical compositions comprising the crystalline polymorph a and at least one excipient or carrier.

Crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is used for the treatment or prevention of cancer or inflammatory diseases. The present invention further relates to a method of treating or preventing cancer or inflammatory diseases comprising administering to a subject in need thereof an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. Yet other aspects of the present invention relate to methods of treating or preventing an inflammatory disease comprising administering to a subject in need thereof an effective amount of a crystalline polymorph. A further aspect of the invention relates to a method of treating or preventing a proliferative disease comprising administering to a subject in need thereof an effective amount of a crystalline polymorph.

Prodrugs of compounds of formula I

Also described herein are prodrugs of the compounds of formula I and methods of treating disorders. For example, the present invention provides methods of treating diseases by administering prodrugs of compounds of formula I. Prodrugs of the compounds of formula I may be administered in the form of pharmaceutical compositions.

Prodrugs are generally precursors of drugs which, after administration to a patient and subsequent absorption, are converted to active or more active species by some process, such as conversion by metabolic pathways. Some prodrugs have chemical groups on the prodrug that render the prodrug less active and/or impart solubility or some other property to the drug. When the chemical group is cleaved and/or modified from the prodrug, an active drug is produced. All of them are generally useful because in some cases the prodrug may be easier to administer than the parent drug. For example, a prodrug may be bioavailable by oral administration, whereas its parent is not. The prodrug may also have better solubility in pharmaceutical compositions than the parent drug. A non-limiting example of a prodrug would be a compound as described herein, which is administered as an ester ("prodrug") to facilitate transport across the cell membrane where water solubility is detrimental to mobility, but which is metabolically hydrolyzed to the carboxylic acid active entity once it enters the cell where water solubility is beneficial. Other examples of prodrugs may be short peptides (polyamino acids) bonded to acid groups where the peptides are metabolized to release the active moiety.

To be used as a modifier to facilitate drug delivery to site-specific tissues, prodrugs can be designed to be reversible drug derivatives. Prodrugs have been designed to increase the effective aqueous solubility of the therapeutic compound to target regions where water is the primary solvent, to date. See, e.g., Fedorak et al, am.J.Physiol.,269: G210-218 (1995); McLoed et al, Gastroenterol,106: 405-; hochhaus et al, biomed.Chrom, 6:283-286 (1992); larsen and h.bundgaard, int.j.pharmaceuticals, 37,87 (1987); J.Larsen et al, int.J.pharmaceuticals, 47,103 (1988); sinkula et al, J.Pharm.Sci.,64:181-210 (1975); higuchi and v.stella, Pro-drugs as Novel Delivery Systems, volume 14 of the a.c.s. conference series; and Edward B.Roche, Bioreversible Carriers in Drug Design, American pharmaceutical Association and Pergamon Press,1987, incorporated herein in their entirety.

In addition, prodrug derivatives of the compounds as described herein can be prepared by methods known to those skilled in the art (see, e.g., Saulnier et al (1994), Bioorganic and Medicinal chemistry letters, Vol.4, p.1985 for more details). By way of example only, suitable prodrugs may be prepared by reacting a non-derivatized compound of formula I with a suitable carbamoylating agent such as, but not limited to, 1-acyloxyalkyl chloroformate, p-nitrophenyl carbonate, and the like. Prodrug forms of the compounds described herein, wherein the prodrug is metabolized in vivo to produce a derivative as described herein, are included within the scope of the claims. Indeed, some of the compounds described herein may be a prodrug of another derivative or active compound.

In some embodiments, prodrugs include compounds in which an amino acid residue or a polypeptide chain of two or more (e.g., 2, 3, or 4) amino acid residues is covalently linked through an amide or ester bond to a free amino, hydroxyl, or carboxylic acid group of a compound of the invention. The amino acid residues include, but are not limited to, the 20 naturally occurring amino acids commonly referred to by three letter symbols, and also include 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, 3-methylhistidine, norvaline, β -alanine, γ -aminobutyric acid, citrulline, homocysteine, homoserine, ornithine and methionine sulfone. Other types of prodrugs are also included.

Compounds of formula I having a free amino, hydroxyl or carboxyl group may be converted into prodrugs. For example, free carboxyl groups may be derivatized to amides or alkyl esters. As outlined in Advanced Drug Delivery Reviews 1996,19,115, free hydroxyl groups can be derivatized to groups including, but not limited to, hemisuccinate, phosphate, dimethylaminoacetate, and phosphoryloxymethyloxycarbonyl. Carbamate prodrugs of hydroxy and amino groups are also included, as are carbonate prodrugs, sulfonates, and sulfates of hydroxy groups.

Also included are derivatization of hydroxyl groups into (acyloxy) methyl ethers and (acyloxy) ethyl ethers, wherein the acyl group may be an alkyl ester, optionally substituted with groups including, but not limited to, ether, amine, and carboxylic acid functional groups, or wherein the acyl group is an amino acid ester as described above. Such prodrugs are described in j.med.chem.1996,39, 10. The free amine may also be derivatized as an amide, sulfonamide, or phosphoramide. All of these prodrug moieties may incorporate groups including, but not limited to, ether, amine, or carboxylic acid functional groups.

Sites on the aromatic ring portion of the compounds of formula I may be susceptible to various metabolic reactions, and thus the introduction of appropriate substituents on the aromatic ring structure may reduce, minimize or eliminate this metabolic pathway.

Pharmaceutical composition

Pharmaceutical compositions are described herein. In some embodiments, the pharmaceutical composition comprises an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition comprises an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is for treating a disorder. In some embodiments, the pharmaceutical composition is for treating a disorder in a mammal. In some embodiments, the pharmaceutical composition is for treating a disorder in a human.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. Such compositions may contain adjuvants, excipients and preservatives, agents for delaying absorption, fillers, binders, adsorbents, buffers, disintegrants, solubilizers, other carriers, and other inert ingredients. Methods of formulating such compositions are well known in the art.

In some embodiments, the pharmaceutical composition is in a form suitable for oral administration. In further or other embodiments, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release formulation, solution, suspension, sterile solution, suspension or emulsion for parenteral injection, ointment or cream for topical administration, or suppository for rectal administration.

In further or other embodiments, the pharmaceutical composition is in unit dosage form suitable for single administration of an accurate dose. In further or other embodiments, the amount of the compound of formula I ranges from about 0.001 to about 1000mg/kg body weight/day. In further or other embodiments, the amount of the compound of formula I is in the range of about 0.5 to about 50 mg/kg/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.001 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.01 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.02 to about 5 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.05 to about 2.5 g/day. In further or other embodiments, the amount of the compound of formula I is from about 0.1 to about 1 g/day. In further or alternative embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate. In further or alternative embodiments, dosage levels above the upper limit of the range recited above may be necessary.

In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the pharmaceutical composition is for administration to a mammal. In further or other embodiments, the mammal is a human.

In further or other embodiments, the pharmaceutical composition further comprises a pharmaceutical carrier, excipient and/or adjuvant. In further or other embodiments, the pharmaceutical composition further comprises at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins, antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal/anti-hormonal therapy agents, and hematopoietic growth factors. In further or other embodiments, the therapeutic agent is paclitaxel, bortezomib, or both. In further or other embodiments, the pharmaceutical composition is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable salt of the compound of formula I.

The invention also relates to a composition comprisingThe composition of (1). In some embodiments, the 2-OH carbon on the compound is in the R configuration. In some embodiments, the composition is substantially free of the S-isomer of the compound. In some embodiments, the compound comprises less than 10% of the S-isomer of the compound. In some embodiments, the compound comprises less than 5% of the S-isomer of the compoundAnd (3) a body. In some embodiments, the compound comprises less than 1% of the S-isomer of the compound. In some embodiments, the compound is in the R configuration.

In some embodiments, the 2-OH carbon on the compound is in the S configuration. In some embodiments, the composition is substantially free of the R-isomer of the compound. In some embodiments, the compound comprises less than 10% of the R-isomer of the compound. In some embodiments, the compound comprises less than 5% of the R-isomer of the compound. In some embodiments, the compound comprises less than 1% of the R-isomer of the compound. In some embodiments, the compound is in the S configuration.

In some embodiments, the composition comprises at least about 50% of the compound exhibiting a powder X-ray diffraction pattern comprising at least 50% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 70% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 90% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5.

In some embodiments, the composition comprises at least about 75% of the compound exhibiting a powder X-ray diffraction pattern comprising at least 50% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 70% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 90% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5.

In some embodiments, the composition comprises at least about 90% of the compound exhibiting a powder X-ray diffraction pattern comprising at least 50% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 70% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 90% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5.

In some embodiments, substantially all of the compound in the composition exhibits a powder X-ray diffraction pattern comprising at least 50% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 70% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern comprises at least 90% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5. In some embodiments, the powder X-ray diffraction pattern is substantially the same as the powder X-ray diffraction pattern shown in fig. 5.

In some embodiments, the crystalline polymorph present in the composition has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph is substantially free of solvent.

In some embodiments, the composition comprises at least about 50% of a compound that exhibits a differential scanning calorimetry pattern substantially the same as the differential scanning calorimetry pattern shown in figure 6. In some embodiments, the crystalline polymorph has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph is substantially free of solvent.

In some embodiments, the composition comprises at least about 75% of a compound that exhibits a differential scanning calorimetry pattern substantially the same as the differential scanning calorimetry pattern shown in figure 6. In some embodiments, the crystalline polymorph has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph is substantially free of solvent.

In some embodiments, the composition comprises at least about 90% of a compound that exhibits a differential scanning calorimetry pattern substantially the same as the differential scanning calorimetry pattern shown in figure 6. In some embodiments, the crystalline polymorph has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph is substantially free of solvent.

In some embodiments, substantially all of the compound in the composition exhibits a differential scanning calorimetry pattern substantially the same as the differential scanning calorimetry pattern shown in figure 6. In some embodiments, the crystalline polymorph has a melting point onset of about 143 ℃ as determined by differential scanning calorimetry. In some embodiments, the crystalline polymorph is substantially anhydrous. In some embodiments, the crystalline polymorph is substantially free of solvent.

In some embodiments, the polymorph of N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is prepared by a method comprising the step of crystallizing amorphous N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. In some embodiments, the crystallizing step comprises crystallizing from a mixture of ethyl acetate and heptane, for example, in a ratio of about 1-4 parts ethyl acetate to about 2-10 parts heptane, or more specifically, in a ratio of about 2 parts ethyl acetate to about 5 parts heptane.

In some embodiments, the compound is prepared as a formulation that directly releases the compound. In some embodiments, the compound is formulated to provide sustained release of the compound. In other embodiments, the compound is prepared as a formulation that delays release of the compound.

In some embodiments, the composition is in a tablet dosage form. In other embodiments, the composition is in the form of a capsule. The compositions may be prepared in capsule or tablet dosage forms, and a wide range of alternative compositions and methods of preparation may be used, references: (1) remington, The Science and Practice of Pharmacy, 20 th edition, 2000, (2) Pharmaceutical document Forms Tablets, volume 1-3, 1989, and (3) modern pharmaceuticals, 4 th edition, 2002. A range of manufacturing processes can be used including dry blending, wet granulation, roller compaction, extrusion, spheronization, coating and spray drying. Soft gelatin formulations and methods of preparation are also suitable.

In some embodiments, the composition comprises a filler or diluent. In various embodiments, the filler or diluent is selected from the group consisting of microcrystalline cellulose, silicified microcrystalline cellulose, lactose, mannitol, compressible sugar, calcium phosphate, calcium sulfate, calcium carbonate, calcium silicate, and starch. In other embodiments, the filler or diluent is microcrystalline cellulose.

In some embodiments, the composition comprises a disintegrant. In various embodiments, the disintegrant is selected from the group consisting of croscarmellose sodium, sodium starch glycolate, crospovidone, methylcellulose, alginic acid, sodium alginate, starch derivatives, bentonite, and magnesium aluminum silicate. In a certain embodiment, the disintegrant is croscarmellose sodium.

In some embodiments, the composition comprises a lubricant. In various embodiments, the lubricant is selected from magnesium stearate, metallic stearates, talc, sodium stearyl fumarate, and stearic acid. In some embodiments, the lubricant is magnesium stearate.

In some embodiments, the composition comprises a wetting agent or surfactant. In various embodiments, the wetting agent or surfactant is selected from sodium lauryl sulfate, glycerin, sorbitan oleate, sorbitan stearate, polyoxyethylated sorbitan laurate, palmitate, stearate, oleate or hexaoleate, polyoxyethylene stearate, and sorbitan monolaurate. In some embodiments, the wetting agent or surfactant is sodium lauryl sulfate.

Other excipients such as glidants, flavoring agents and coloring agents may also be added. Other optional Excipients can be found in The Handbook of pharmaceutical Excipients, 5 th edition, 2005 and FDA inactive ingredients database.

The present invention also relates to a composition comprising:

about 1mg of a structureA compound of (a) (as defined in any of the above embodiments);

about 222.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 10mg of structureA compound of (a) (as defined in any of the above embodiments);

about 213.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 20mg of the structureA compound of (a) (as defined in any of the above embodiments);

about 203.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 40mg of the structureA compound of (a) (as defined in any of the above embodiments);

About 183.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising: about 0.4 wt% of the structure isA compound of (a) (as defined in any of the above embodiments); and about 99.6% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrierThe body or carrier comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises about 92.6% by weight of the composition. In further or other embodiments, the composition further comprises: about 5% by weight croscarmellose sodium; about 1% by weight sodium lauryl sulfate; and about 1% by weight magnesium stearate.

The present invention also relates to a composition comprising: about 4.2 wt% of the structure isA compound of (a) (as defined in any of the above embodiments); and about 95.8% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises about 88.8% by weight of the composition. In further or other embodiments, the composition further comprises: about 5% by weight croscarmellose sodium; about 1% by weight sodium lauryl sulfate; and about 1% by weight magnesium stearate.

The present invention also relates to a composition comprising: about 2% to about 10% by weight of structures ofA compound of (a) (as defined in any of the above embodiments); and about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises from about 85% to about 95% by weight of the composition. In further or other embodiments, the composition further comprises: from about 1% to about 6% by weight of croscarmellose sodium; about 0.1% to about 2% by weight sodium lauryl sulfate; and about 0.25% to about 1.5% by weight of magnesium stearate. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline celluloseFrom about 85% to about 95% by weight of the composition. In further or other embodiments, the composition further comprises: from about 1% to about 6% by weight of croscarmellose sodium; and about 0.25% to about 1.5% by weight of magnesium stearate.

The present invention also relates to a composition comprising:

about 1mg of a structureA compound of (1);

about 222.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 10mg of structureA compound of (1);

about 213.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 20mg of the structureA compound of (1);

about 203.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 40mg of the structureA compound of (1);

about 183.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising: about 0.4 wt% of the structure isA compound of (1); and about 99.6% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises about 92.6% by weight of the composition. In further or other embodiments, the composition further comprises: about 5% by weight croscarmellose sodium; about 1% by weight sodium lauryl sulfate; and about 1% by weight magnesium stearate.

The present invention also relates to a composition comprising: about 4.2 wt% of the structure isA compound of (1); and about 95.8% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises the compositionAbout 88.8 wt%. In further or other embodiments, the composition further comprises: about 5% by weight croscarmellose sodium; about 1% by weight sodium lauryl sulfate; and about 1% by weight magnesium stearate.

The present invention also relates to a composition comprising: about 2% to about 10% by weight of structures ofA compound of (1); and about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises from about 85% to about 95% by weight of the composition. In further or other embodiments, the composition further comprises: from about 1% to about 6% by weight of croscarmellose sodium; about 0.1% to about 2% by weight sodium lauryl sulfate; and about 0.25% to about 1.5% by weight of magnesium stearate. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises from about 85% to about 95% by weight of the composition. In further or other embodiments, the composition further comprises: from about 1% to about 6% by weight of croscarmellose sodium; and about 0.25% to about 1.5% by weight of magnesium stearate.

The present invention also relates to a composition comprising:

about 1mg of a structureA compound of (1);

about 222.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 10mg of structureA compound of (1);

about 213.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 20mg of the structureA compound of (1);

about 203.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising:

about 40mg of the structureA compound of (1);

about 183.2mg microcrystalline cellulose;

about 12.0mg croscarmellose sodium;

about 2.4mg sodium lauryl sulfate; and

about 2.4mg magnesium stearate.

The present invention also relates to a composition comprising: about 4.2 wt% of the structure isA compound of (1); and about 95.8% by weight of a pharmaceutically acceptable carrier or vehicle. In some embodiments, the pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In further or other embodiments, the microcrystalline cellulose comprises about 88.8% by weight of the composition. In further or other embodiments, the composition further comprises: about 5% by weight croscarmellose sodium; about 1% by weight sodium lauryl sulfate; and about 1% by weight magnesium stearate.

Also described herein are pharmaceutical compositions comprising an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. In some embodiments, the pharmaceutical composition comprises an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is for treating a disorder. In some embodiments, the pharmaceutical composition is for treating a disorder in a mammal. In some embodiments, the pharmaceutical composition is for treating a disorder in a human. In some embodiments, the pharmaceutical composition is for treating or preventing an inflammatory disease. In some embodiments, the pharmaceutical composition is for treating or preventing a proliferative disease.

Methods of use of the compounds and compositions, including polymorphs

In other aspects, the invention relates to a method of achieving an effect in a patient comprising administering to the patient an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer or prodrug thereof, wherein the effect is selected from the group consisting of inhibition of various cancers, immune diseases and inflammatory diseases. In some embodiments, the compound, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or vehicle. In some embodiments, the effect is inhibition of various cancers. In further or other embodiments, the effect is suppression of an immune disease. In further or other embodiments, the effect is inhibition of an inflammatory disease.

Any of the compositions described and claimed herein can be used in the methods provided in this section.

In some embodiments, a composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, or surgery, or any combination thereof. In further or other embodiments, a composition comprising a compound of formula I is administered in combination with at least one therapeutic agent.

In some embodiments of the compositions and methods provided herein, further packets are providedA MEK protein kinase inhibitor comprising a compound of formula I wherein the compound of formula I is present in an amount from about 0.1m to about 200 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 0.2mg to about 100 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 0.3mg to about 90 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 0.4mg to about 80 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 0.5mg to about 70 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 0.4mg to about 80 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 0.5mg to about 70 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount of from about 1mg to about 60 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 1.5mg to about 50 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount of from about 2mg to about 45 mg. In other embodiments, the MEK protein kinase inhibitor comprises a compound of formula I, and is present in an amount from about 2.5mg to about 40 mg. In a further embodiment, MEK protein kinase inhibitors further comprising a compound of formula I in the dosages provided herein are selected from

In some embodiments of the compositions and methods provided herein, MEK protein kinase inhibitors are provided that further comprise a compound of formula I, wherein the compound of formula I is present in an amount of about 0.1mg, about 0.2mg, about 0.25mg, about 0.3mg, about 0.4mg, about 0.5mg, about 0.6mg, about 0.7mg, about 0.8mg, about 0.9mg, about 1mg, about 1.5mg, about 2mg, about 2.5mg, about 3mg, about 3.5mg, about 4.0mg, about 4.5mg, about 5mg, about 5.5 mgmg, about 6mg, about 6.5mg, about 7mg, about 7.5mg, about 8mg, about 8.5mg, about 9mg, about 9.5mg, about 10mg, about 10.5mg, about 11mg, about 11.5mg, about 12mg, about 12.5mg, and/or about 13mg, about 14mg, or about 15 mg. In a further embodiment, the compound of formula I present in the doses provided herein is selected from

In some embodiments of the compositions and methods provided herein, a MEK protein kinase inhibitor is provided that further comprises a compound of formula I, wherein the compound of formula I is present in an amount of about 15mg, about 20mg, about 25mg, about 30mg, about 35mg, about 40mg, about 45mg, about 50mg, about 55mg, about 60mg, about 65mg, about 75mg, about 80mg, about 85mg, about 90mg, about 95mg, about 100mg, about 110mg, about 120mg, about 125mg, about 130mg, about 140mg, about 150mg, about 160mg, about 170mg, about 175mg, about 180mg, about 190mg, or about 200 mg. In a further embodiment, the compound of formula I present in the doses provided herein is selected from

In some aspects, the invention relates to a method of treating a disease in a subject suffering from said disease, comprising administering to said subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof.

Diseases and disorders modulated by MEK

Also described herein are methods of modulating MEK activity by contacting MEK with an amount of a compound of formula I sufficient to modulate MEK activity. Modulation may be inhibition or activation of MEK activity. In some embodiments, the present invention provides methods of inhibiting MEK activity by contacting MEK with an amount of a compound of formula I sufficient to inhibit MEK activity. In some embodiments, the present invention provides methods of inhibiting MEK activity in a solution by contacting the solution with an amount of a compound of formula I sufficient to inhibit MEK activity in the solution. In some embodiments, the present invention provides methods of inhibiting MEK activity in a cell by contacting the cell with an amount of a compound described herein sufficient to inhibit MEK activity in the cell. In some embodiments, the present invention provides methods of inhibiting MEK activity in a tissue by contacting the tissue with an amount of a compound described herein sufficient to inhibit MEK activity in the tissue. In some embodiments, the present invention provides methods of inhibiting MEK activity in an organism by contacting the organism with an amount of a compound described herein sufficient to inhibit MEK activity in the organism. In some embodiments, the present invention provides methods of inhibiting MEK activity in an animal by contacting the animal with an amount of a compound described herein sufficient to inhibit MEK activity in the animal. In some embodiments, the present invention provides methods of inhibiting MEK activity in a mammal by contacting the mammal with an amount of a compound described herein sufficient to inhibit MEK activity in the mammal. In some embodiments, the present invention provides methods of inhibiting MEK activity in a human by contacting the human with an amount of a compound described herein sufficient to inhibit MEK activity in the human.

Compounds of formula I and compositions comprising compounds of formula I, and pharmaceutically acceptable salts, solvates, polymorphs, esters, amides, tautomers, or prodrugs thereof, modulate the activity of MEK enzymes; and as such are used to treat diseases or conditions in which abnormal MEK enzyme activity results in a pathology and/or symptom of the disease or condition.

In some aspects, the invention relates to methods of treating a disorder or condition modulated by the MEK cascade in a mammal, including a human, comprising administering to said mammal an amount of a compound of formula I, or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof, effective to modulate said cascade. The appropriate dosage for a particular patient can be determined by one skilled in the art according to known methods.

In other aspects, the invention relates to methods of inhibiting MEK enzymes. In some embodiments, the methods comprise contacting the MEK enzyme with an amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, sufficient to inhibit the enzyme, wherein the enzyme is inhibited. In further or other embodiments, the enzyme is inhibited by at least about 1%. In further or other embodiments, the enzyme is inhibited by at least about 2%. In further or other embodiments, the enzyme is inhibited by at least about 3%. In further or other embodiments, the enzyme is inhibited by at least about 4%. In further or other embodiments, the enzyme is inhibited by at least about 5%. In further or other embodiments, the enzyme is inhibited by at least about 10%. In further or other embodiments, the enzyme is inhibited by at least about 20%. In further or other embodiments, the enzyme is inhibited by at least about 25%. In further or other embodiments, the enzyme is inhibited by at least about 30%. In further or other embodiments, the enzyme is inhibited by at least about 40%. In further or other embodiments, the enzyme is inhibited by at least about 50%. In further or other embodiments, the enzyme is inhibited by at least about 60%. In further or other embodiments, the enzyme is inhibited by at least about 70%. In further or other embodiments, the enzyme is inhibited by at least about 75%. In further or other embodiments, the enzyme is inhibited by at least about 80%. In further or other embodiments, the enzyme is inhibited by at least about 90%. In further or other embodiments, the enzyme is substantially completely inhibited. In further or other embodiments, the MEK enzyme is a MEK kinase. In further or other embodiments, the MEK enzyme is MEK 1. In further or other embodiments, the MEK enzyme is MEK 2. In further or other embodiments, the contacting occurs intracellularly. In further or other embodiments, the cell is a mammalian cell. In further or other embodiments, the mammalian cell is a human cell. In further or other embodiments, the MEK enzyme is inhibited by a composition comprising a pharmaceutically acceptable salt of a compound of formula I.

In a further or other aspect, the present invention relates to a method of treating a MEK-mediated disorder in a subject suffering from such a disorder, comprising administering to the subject an effective amount of a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof. In some embodiments, the composition comprising a compound of formula I is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically, or rectally. In some embodiments, the pharmaceutical composition is in a form suitable for oral administration. In further or other embodiments, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release formulation, solution, suspension, sterile solution, suspension or emulsion for parenteral injection, ointment or cream for topical administration, or suppository for rectal administration. In further or other embodiments, the pharmaceutical composition is in unit dosage form suitable for single administration of an accurate dose. In further or other embodiments, the pharmaceutical composition further comprises a pharmaceutical carrier, excipient and/or adjuvant.

In some embodiments, a composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins, antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal/anti-hormonal therapy agents, and hematopoietic growth factors. In further or other embodiments, the therapeutic agent is selected from paclitaxel, bortezomib, or both.

In some embodiments, the MEK mediated disorder is selected from inflammatory diseases, infections, autoimmune diseases, stroke, ischemia, cardiac disorders, neurological disorders, fibrotic disorders, proliferative disorders, hyperproliferative disorders, non-cancer hyperproliferative disorders, tumors, leukemias, tumors, cancers, metabolic diseases, malignant diseases, vascular restenosis, psoriasis, atherosclerosis, rheumatoid arthritis, osteoarthritis, heart failure, chronic pain, neuropathic pain, dry eye, angle-closure glaucoma and open angle glaucoma. In further or other embodiments, the MEK mediated disorder is an inflammatory disease. In further or other embodiments, the MEK mediated disorder is a hyperproliferative disease. In further or other embodiments, the MEK mediated disorder is selected from a tumor, a leukemia, a neoplasm, a cancer, a carcinoma and a malignant disease. In further or other embodiments, the disorder is gastric cancer, brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, colorectal cancer, or leukemia. In further or other embodiments, the fibrotic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid formation, interstitial nephritis, or pulmonary fibrosis. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

The present invention also relates to methods of modulating MEK activity by contacting MEK with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to modulate MEK activity. Modulation may be inhibition or activation of MEK activity. In some embodiments, the present invention provides methods of inhibiting MEK activity by contacting MEK with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity. In some embodiments, the present invention provides methods of inhibiting MEK activity in a solution by contacting the solution with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the solution. In some embodiments, the present invention provides methods of inhibiting MEK activity in a cell by contacting the cell with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the cell. In some embodiments, the present invention provides methods of inhibiting MEK activity in tissue by contacting the tissue with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the tissue. In some embodiments, the present invention provides methods of inhibiting MEK activity in an organism by contacting the organism with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the organism. In some embodiments, the present invention provides methods of inhibiting MEK activity in an animal by contacting the animal with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the animal. In some embodiments, the present invention provides methods of inhibiting MEK activity in a mammal by contacting the mammal with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the mammal. In some embodiments, the present invention provides methods of inhibiting MEK activity in a human by contacting the human with an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide sufficient to inhibit MEK activity in the human.

Cancer treatment

In other aspects, the present invention relates to methods of treating or preventing cancer in a subject (prevention or prophyylaxis) comprising administering to the subject an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof. In some embodiments, the compound, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or vehicle. In further or other embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, colorectal cancer, gastric cancer, or leukemia. In further or other embodiments, the fibrotic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid formation, interstitial nephritis, or pulmonary fibrosis. In further or other embodiments, the cancer is brain, breast, lung, ovarian, pancreatic, prostate, renal, colorectal, leukemia, melanoma, thyroid, or basal cell carcinoma. In further or other embodiments, the cancer is brain cancer or adrenocortical cancer. In further or other embodiments, the cancer is breast cancer. In further or other embodiments, the cancer is ovarian cancer. In further or other embodiments, the cancer is pancreatic cancer. In further or other embodiments, the cancer is prostate cancer. In further or other embodiments, the cancer is renal cancer. In further or other embodiments, the cancer is colorectal cancer. In further or other embodiments, the cancer is myeloid leukemia. In further or other embodiments, the cancer is glioblastoma. In further or other embodiments, the cancer is follicular lymphoma. In further or other embodiments, the cancer is pre-B acute leukemia. In further or other embodiments, the cancer is chronic lymphocytic B leukemia. In further or other embodiments, the cancer is mesothelioma. In further or other embodiments, the cancer is small cell lung cancer. In some embodiments, the cancer is gastric cancer.

In some embodiments, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly, or by infusion), topically, or rectally. In further or other embodiments, the compound of formula I is administered in a single dose once daily. In further or other embodiments, the compound of formula I is administered in multiple doses more than once daily. In further or other embodiments, the compound of formula I is administered twice daily. In further or other embodiments, the compound of formula I is administered three times daily. In further or other embodiments, the compound of formula I is administered four times daily. In further or other embodiments, the compound of formula I is administered more than four times daily. In some embodiments, the individual having cancer is a mammal. In further or other embodiments, the subject is a human. In further or other embodiments, an effective amount of a composition comprising a pharmaceutically acceptable salt of a compound of formula I is administered.

Abnormal cell growth

Also described herein are compounds, pharmaceutical compositions and methods for inhibiting abnormal cell growth. In some embodiments, the abnormal cell growth occurs in a mammal. A method of inhibiting abnormal cell growth comprises administering an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, wherein abnormal cell growth is inhibited. A method of inhibiting abnormal cell growth in a mammal comprises administering to the mammal an amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, wherein the amount of the compound or salt is effective to inhibit abnormal cell growth in the mammal.

In some embodiments, the method comprises administering an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, in combination with an amount of a chemotherapeutic agent, wherein the amount of the compound, or salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, and the chemotherapeutic agent together are effective to inhibit abnormal cell growth. Many chemotherapeutic agents are currently known in the art and may be used in combination with the compounds of the present invention. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, angiogenesis inhibitors, and anti-androgens.

Also described is a method of inhibiting abnormal cell growth in a mammal, comprising administering to the mammal an amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, in combination with radiation therapy, wherein the amount of the compound, or the salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, in combination with radiation therapy is effective to inhibit abnormal cell growth in the mammal or to treat a hyperproliferative disease in the mammal. Techniques for administering radiation therapy are known in the art, and these techniques may be used in combination with the therapies described herein. Administration of a compound of formula I in this combination therapy can be determined as described herein.

The present invention also relates to methods and pharmaceutical compositions for inhibiting abnormal cell growth in a mammal comprising an amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, or an isotopically labeled derivative thereof, and an amount of one or more substances selected from the group consisting of anti-angiogenic agents, signal transduction inhibitors, and antiproliferative agents.

Anti-angiogenic agents, such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors, may be used in combination with the compounds of the present invention and the pharmaceutical compositions described herein. Examples of useful COX-II inhibitors include: CELEBREX^TM(celecoxib), valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96/33172 (published at 24.10.1996), WO 96/27583 (published at 7.3.1996), European patent application 97304971.1 (published at 8.7.1997), European patent application 99308617.2 (published at 29.10.1999), WO 98/07697 (published at 26.2.1998), WO 98/03516 (published at 29.1.1998), WO 98/34918 (published at 13.8.1998), WO 98/34915 (published at 13.8.1998), WO 98/33768 (published at 6.8.1998), WO 98/30566 (published at 16.7.1998), European patent publication 606,046 (published at 13.7.1994), European patent publication 931,788 (published at 28.7.1999), WO 90/05719 (published at 31.5.1990), WO 99/52910 (published on 21/10/1999), WO 99/52889 (published on 21/10/1999), WO 99/29667 (published on 17/6/1999), PCT International Application PCT/IB98/01113 (filed on 21/7/1998), European patent application 99302232.1 (filed on 25/3/1999), British patent application 9912961.1 (filed on 3/1999), U.S. provisional application 60/148,464 (filed on 12/8/1999), U.S. patent 5,863,949 (filed on 26/1999), U.S. patent 5,861,510 (filed on 19/1999), and European patent publication 780,386 (filed on 25/6/1997), all of which are incorporated herein by reference in their entirety. Some MMP-2 and MMP-9 inhibitors have little or no MMP-1 inhibitory activity, while some MMP-2 and MMP-9 inhibitors selectively inhibit MMP-2 and/or AMP-9 relative to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-ll, MMP-12, and MMP-13). Some specific examples of MMP inhibitors useful in the present invention are AG-3340, RO 32-3555, and RS 13-0830.

In other aspects, the invention relates to a method of degenerating, inhibiting the growth of, or killing a cancer cell comprising contacting the cell with a composition comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, in an amount effective to degenerate, inhibit the growth of, or kill the cell. In some embodiments, the cancer cell comprises a brain cancer cell, a breast cancer cell, a lung cancer cell, an ovarian cancer cell, a pancreatic cancer cell, a prostate cancer cell, a renal cancer cell, or a colorectal cancer cell.

In further or other embodiments, the composition is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is paclitaxel, bortezomib, or both. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins; antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormone/anti-hormone therapeutics, and hematopoietic growth factors.

In some embodiments, the cancer cell is degenerated. In further or other embodiments, 1% of the cancer cells are degenerated. In further or other embodiments, 2% of the cancer cells are degenerated. In further or other embodiments, 3% of the cancer cells are degenerated. In further or other embodiments, 4% of the cancer cells are degenerated. In further or other embodiments, 5% of the cancer cells are degenerated. In further or other embodiments, 10% of the cancer cells are degenerated. In further or other embodiments, 20% of the cancer cells are degenerated. In further or other embodiments, 25% of the cancer cells are degenerated. In further or other embodiments, 30% of the cancer cells are degenerated. In further or other embodiments, 40% of the cancer cells are degenerated. In further or other embodiments, 50% of the cancer cells are degenerated. In further or other embodiments, 60% of the cancer cells are degenerated. In further or other embodiments, 70% of the cancer cells are degenerated. In further or other embodiments, 75% of the cancer cells are degenerated. In further or other embodiments, 80% of the cancer cells are degenerated. In further or other embodiments, 90% of the cancer cells are degenerated. In further or other embodiments, 100% of the cancer cells are degenerated. In further or other embodiments, substantially all of the cancer cells are degenerated.

In some embodiments, the cancer cells are killed. In further or other embodiments, 1% of the cancer cells are killed. In further or other embodiments, 2% of the cancer cells are killed. In further or other embodiments, 3% of the cancer cells are killed. In further or other embodiments, 4% of the cancer cells are killed. In further or other embodiments, 5% of the cancer cells are killed. In further or other embodiments, 10% of the cancer cells are killed. In further or other embodiments, 20% of the cancer cells are killed. In further or other embodiments, 25% of the cancer cells are killed. In further or other embodiments, 30% of the cancer cells are killed. In further or other embodiments, 40% of the cancer cells are killed. In further or other embodiments, 50% of the cancer cells are killed. In further or other embodiments, 60% of the cancer cells are killed. In further or other embodiments, 70% of the cancer cells are killed. In further or other embodiments, 75% of the cancer cells are killed. In further or other embodiments, 80% of the cancer cells are killed. In further or other embodiments, 90% of the cancer cells are killed. In further or other embodiments, 100% of the cancer cells are killed. In further or other embodiments, substantially all of the cancer cells are killed.

In further or other embodiments, the growth of the cancer cell is inhibited. In further or other embodiments, the growth of the cancer cells is inhibited by about 1%. In further or other embodiments, the growth of the cancer cells is inhibited by about 2%. In further or other embodiments, the growth of the cancer cells is inhibited by about 3%. In further or other embodiments, the growth of the cancer cells is inhibited by about 4%. In further or other embodiments, the growth of the cancer cells is inhibited by about 5%. In further or other embodiments, the growth of the cancer cells is inhibited by about 10%. In further or other embodiments, the growth of the cancer cells is inhibited by about 20%. In further or other embodiments, the growth of the cancer cells is inhibited by about 25%. In further or other embodiments, the growth of the cancer cells is inhibited by about 30%. In further or other embodiments, the growth of the cancer cells is inhibited by about 40%. In further or other embodiments, the growth of the cancer cells is inhibited by about 50%. In further or other embodiments, the growth of the cancer cells is inhibited by about 60%. In further or other embodiments, the growth of the cancer cells is inhibited by about 70%. In further or other embodiments, the growth of the cancer cells is inhibited by about 75%. In further or other embodiments, the growth of the cancer cells is inhibited by about 80%. In further or other embodiments, the growth of the cancer cells is inhibited by about 90%. In further or other embodiments, the growth of the cancer cells is inhibited by about 100%. In further or other embodiments, compositions comprising pharmaceutically acceptable salts of compounds of formula I are used.

Also described herein are methods of inhibiting abnormal cell growth. In some embodiments, the abnormal cell growth occurs in a mammal. A method of inhibiting abnormal cell growth comprising administering an effective amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, wherein abnormal cell growth is inhibited. A method of inhibiting abnormal cell growth in a mammal comprising administering to said mammal an amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, wherein the amount of crystalline polymorph a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is effective to inhibit abnormal cell growth in the mammal.

In some embodiments, the methods comprise administering an effective amount of a crystalline polymorph A of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide in combination with an amount of a chemotherapeutic agent, wherein the crystalline polymorph A of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and the chemotherapeutic agent are in amounts that together are effective to inhibit abnormal cell growth.

Currently, a number of chemotherapeutic agents are known in the art and may be combined with the compounds and compositions of the present invention. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, angiogenesis inhibitors, and anti-androgens.

In some embodiments, a method of inhibiting abnormal cell growth in a mammal comprises administering to said mammal an amount of crystalline polymorph A of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide in combination with radiation therapy, wherein the amount of crystalline polymorph A of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is effective to inhibit abnormal cell growth in combination with the radiation therapy. Techniques for administering radiation therapy are known in the art, and these techniques may be used in the combination therapies described herein.

Treatment of hyperproliferative disorders

In other aspects, the invention relates to methods of treating hyperproliferative disorders in mammals, including humans, comprising administering to said mammal a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof.

In other aspects, the invention relates to a method of treating or preventing a proliferative disease in a subject, comprising administering to the subject an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer or prodrug thereof. In some embodiments, the compound, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or vehicle. In some embodiments, the proliferative disease is cancer, psoriasis, restenosis, autoimmune disease, or atherosclerosis. In further or other embodiments, the proliferative disease is a hyperproliferative disease. In further or other embodiments, the proliferative disease is selected from the group consisting of a tumor, leukemia, a neoplasm, a cancer, a carcinoma, and a malignant disease. In further or other embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, gastric cancer, colorectal cancer, or leukemia. In further or other embodiments, the fibrotic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid formation, interstitial nephritis, or pulmonary fibrosis. In further or other embodiments, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, gastric cancer, pancreatic cancer, prostate cancer, renal cancer, colorectal cancer, or leukemia. In further or other embodiments, the cancer is brain cancer or adrenocortical cancer. In further or other embodiments, the cancer is breast cancer. In further or other embodiments, the cancer is ovarian cancer. In further or other embodiments, the cancer is pancreatic cancer. In further or other embodiments, the cancer is prostate cancer. In further or other embodiments, the cancer is renal cancer. In further or other embodiments, the cancer is colorectal cancer. In further or other embodiments, the cancer is myeloid leukemia. In further or other embodiments, the cancer is glioblastoma. In further or other embodiments, the cancer is follicular lymphoma. In further or other embodiments, the cancer is pre-B acute leukemia. In further or other embodiments, the cancer is chronic lymphocytic B leukemia. In further or other embodiments, the cancer is mesothelioma. In further or other embodiments, the cancer is small cell lung cancer. In some embodiments, the cancer is gastric cancer.

In some embodiments, a composition comprising a compound of formula I is administered in combination with other therapies. In further or other embodiments, the other therapy is radiation therapy, chemotherapy, surgery, or a combination thereof. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent. In further or other embodiments, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenic agents, and anti-neoplastic agents. In further or other embodiments, the antineoplastic agent is selected from the group consisting of alkylating agents, antimetabolites, epiphyllotoxins, antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal/anti-hormonal therapy agents, and hematopoietic growth factors. In further or other embodiments, the therapeutic agent is selected from paclitaxel, bortezomib, or both.

Size of tumor

In other aspects, the present invention relates to a method of reducing tumor size, inhibiting tumor size increase, reducing tumor proliferation, or preventing tumor proliferation in a subject, comprising administering to the subject an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof. In some embodiments, the compound, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or vehicle. In some embodiments, the size of the tumor is reduced. In further or other embodiments, the size of the tumor is reduced by at least 1%. In further or other embodiments, the size of the tumor is reduced by at least 2%. In further or other embodiments, the size of the tumor is reduced by at least 3%. In further or other embodiments, the size of the tumor is reduced by at least 4%. In further or other embodiments, the size of the tumor is reduced by at least 5%. In further or other embodiments, the size of the tumor is reduced by at least 10%. In further or other embodiments, the size of the tumor is reduced by at least 20%. In further or other embodiments, the size of the tumor is reduced by at least 25%. In further or other embodiments, the size of the tumor is reduced by at least 30%. In further or other embodiments, the size of the tumor is reduced by at least 40%. In further or other embodiments, the size of the tumor is reduced by at least 50%. In further or other embodiments, the size of the tumor is reduced by at least 60%. In further or other embodiments, the size of the tumor is reduced by at least 70%. In further or other embodiments, the size of the tumor is reduced by at least 75%. In further or other embodiments, the size of the tumor is reduced by at least 80%. In further or other embodiments, the size of the tumor is reduced by at least 85%. In further or other embodiments, the size of the tumor is reduced by at least 90%. In further or other embodiments, the size of the tumor is reduced by at least 95%. In further or other embodiments, the tumor is eradicated. In some embodiments, the size of the tumor is not increased.

In some embodiments, tumor proliferation is slowed. In some embodiments, tumor proliferation is reduced by at least 1%. In some embodiments, tumor proliferation is reduced by at least 2%. In some embodiments, tumor proliferation is reduced by at least 3%. In some embodiments, tumor proliferation is reduced by at least 4%. In some embodiments, tumor proliferation is reduced by at least 5%. In some embodiments, tumor proliferation is reduced by at least 10%. In some embodiments, tumor proliferation is reduced by at least 20%. In some embodiments, tumor proliferation is reduced by at least 25%. In some embodiments, tumor proliferation is reduced by at least 30%. In some embodiments, tumor proliferation is reduced by at least 40%. In some embodiments, tumor proliferation is reduced by at least 50%. In some embodiments, tumor proliferation is reduced by at least 60%. In some embodiments, tumor proliferation is reduced by at least 70%. In some embodiments, tumor proliferation is reduced by at least 75%. In some embodiments, tumor proliferation is reduced by at least 75%. In some embodiments, tumor proliferation is reduced by at least 80%. In some embodiments, tumor proliferation is reduced by at least 90%. In some embodiments, tumor proliferation is reduced by at least 95%. In some embodiments, tumor proliferation is prevented.

Inflammatory diseases

In other aspects, the invention relates to a method of treating or preventing an inflammatory disease in a subject, comprising administering to the subject an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer or prodrug thereof. In some embodiments, the compound, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, or prodrug thereof, is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or vehicle. In a further or additional embodiment, the inflammatory disease is selected from chronic inflammatory disease, rheumatoid arthritis, spondyloarthropathies, ankylosing spondylitis, gout, tendonitis, bursitis, sciatica, gouty arthritis, osteoarthritis, juvenile arthritis, acute rheumatic arthritis, enteropathic arthritis, neuropathic arthritis, psoriatic arthritis, suppurative arthritis, atherosclerosis, systemic lupus erythematosus, inflammatory bowel disease, irritable bowel syndrome, ulcerative colitis, reflux esophagitis, crohn's disease, gastritis, asthma, allergy, respiratory distress syndrome, pancreatitis, chronic obstructive pulmonary disease, pulmonary fibrosis, psoriasis, eczema, or scleroderma.

In some embodiments, a composition comprising a compound of formula (la) is administered in combination with other therapies. In further or other embodiments, the composition comprising a compound of formula I is administered in combination with at least one therapeutic agent. In some embodiments, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly, or by infusion), topically, or rectally. In further or other embodiments, the amount of the compound of formula I ranges from about 0.001 to about 1000mg/kg body weight/day. In further or other embodiments, the amount of the compound of formula I is in the range of about 0.5 to about 50 mg/kg/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.001 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.01 to about 7 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.02 to about 5 g/day. In a further or additional embodiment, the amount of the compound of formula I is from about 0.05 to about 2.5 g/day. In further or other embodiments, the amount of the compound of formula I is from about 0.1 to about 1 g/day. In further or alternative embodiments, dosage levels below the lower limit of the aforesaid range may be more than adequate. In further or alternative embodiments, dosage levels above the upper limit of the range recited above may be necessary.

Modes of administration

Described herein are compounds of formula I or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof. Also described are pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof. The compounds and compositions described herein may be administered alone or in combination with pharmaceutically acceptable carriers, excipients, or diluents in pharmaceutical compositions in accordance with standard pharmaceutical practice.

Also described herein are pharmaceutical compositions comprising a crystalline polymorph of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form a). The compounds and compositions described herein may be administered alone or in combination with pharmaceutically acceptable carriers, excipients, or diluents in pharmaceutical compositions in accordance with standard pharmaceutical practice. Administration can be achieved by any method capable of delivering the compound to the site of action. These methods include, but are not limited to, delivery by enteral routes (including oral, gastric or duodenal feeding tubes, rectal suppositories, and rectal enemas), parenteral routes (injection or infusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and subcutaneous), inhalation, transdermal, transmucosal, sublingual, buccal, and topical (including epicutaneous (epicutaneous), dermal, enema, eye drops, ear drops, intranasal, vaginal) administration, although the most appropriate route may depend on, for example, the condition and disorder of the recipient. Those skilled in the art will be familiar with the administration techniques that may be used for the compounds and methods of the present invention. By way of example only, a compound described herein may be administered locally to an area in need of treatment by, for example, local infusion during surgery, local administration such as a cream or ointment, injection, catheter, or implant made of, for example, porous, non-porous, or gelatinous material (including membranes or fibers such as sialastic membranes). Administration can also be by direct injection at the site of the diseased tissue or organ.

Administration of the compounds and compositions described herein can be achieved by any method capable of releasing the compound to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), topical administration and rectal administration. For example, a compound described herein may be administered locally to an area in need of treatment. This may be achieved by, for example, but not limited to, local infusion during surgery, local administration of, for example, a cream, ointment, injection, catheter or implant made of, for example, porous, non-porous or gelatinous material (including membranes or fibers such as sialastic membranes). Administration can also be by direct injection at the site (or original site) of the tumor or neoplastic or pre-neoplastic tissue. Formulations and administration techniques that may be used for The compounds and methods of The present invention are well known to those skilled in The art, for example, in Goodman and Gilman, The pharmaceutical basic of Therapeutics, current edition; pergamon and Remington's, Pharmaceutical Sciences (current edition), Mack publishing Co., Easton, Pa.

The formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intraarticular and intramedullary), intraperitoneal, transmucosal, transdermal, rectal and topical administration (including dermal, buccal, sublingual and intraocular administration), although the most suitable route may depend, for example, on the condition and disorder of the recipient. The formulations may conveniently be presented in unit dosage form and may be formulated by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association a compound of the invention, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof ("active ingredient"), with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired formulation.

Formulations suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; powder or granules; a solution or suspension in an aqueous liquid or a non-aqueous liquid; or an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be a bolus, electuary or paste.

Pharmaceutical formulations for oral administration include tablets, push-fit capsules made of gelatin, and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. Tablets may be prepared by compression or moulding, optionally together with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, inert diluent or lubricant, surfactant or surface dispersant. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therefrom. All formulations for oral administration should be in dosages suitable for such administration. The push-fit capsules or tablets may contain an active ingredient; mixing with fillers such as microcrystalline cellulose, silicified microcrystalline cellulose, pregelatinized starch, lactose, calcium hydrogen phosphate or compressible sugar; binders such as hypromellose, povidone, or starch paste; disintegrants such as croscarmellose sodium, crospovidone or sodium starch glycolate; surfactants such as sodium lauryl sulfate and/or lubricants, and processing aids such as talc, magnesium stearate, stearic acid or colloidal silicon dioxide, and optionally stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. Dragee cores are suitably coated. For this purpose, concentrated sugar solutions are used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to identify or characterize different combinations of active compound doses, dyes or pigments may be added to the tablets or dragee coatings.

The pharmaceutical preparation may be prepared as a preparation for parenteral administration by injection, such as bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents (formulations) such as suspending, stabilizing and/or dispersing agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in powder form or in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example saline or sterile, pyrogen-free water, immediately prior to use. From sterile powders, granules and tablets of the type described above, ready-to-inject solutions and suspensions can be prepared.

Formulations for parenteral administration include aqueous and non-aqueous (oily) sterile injectable solutions of the active compound which may contain anti-oxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions may contain suspending agents and thickening agents. Suitable lipophilic solvents or vehicles include fatty oils, for example sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, to enable the formulation of highly concentrated solutions, the suspensions may also contain suitable stabilizers or agents that increase the solubility of the compounds.

The pharmaceutical formulation may also be prepared as a depot formulation. Such long acting formulations may be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (e.g. as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, e.g. as a sparingly soluble salt.

For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, pastilles or gels prepared in a conventional manner. Such compositions may contain the active ingredient in a flavored base such as sucrose and acacia or tragacanth.

Pharmaceutical formulations may also be prepared as rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter, polyethylene glycols or other glycerides.

The pharmaceutical preparation may be administered locally, i.e. by non-systemic administration. This includes applying the compounds of the invention externally to the epidermis or the oral cavity, as well as instilling such compounds into the ear, eye and nose so that the compounds do not enter the bloodstream in large quantities. By contrast, systemic administration means oral administration, intravenous administration, intraperitoneal administration and intramuscular administration.

Pharmaceutical formulations suitable for topical administration include liquid or semi-liquid formulations suitable for penetration through the skin into the site of inflammation, for example gels, liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. For topical administration, the active ingredient may comprise 0.001% to 10% w/w, for example 1% to 2% by weight of the formulation. However, the active ingredient may comprise up to 10% w/w of the formulation, or may comprise less than 5% w/w, or 0.1% to 1% w/w of the formulation.

Pharmaceutical formulations for administration by inhalation may conveniently be presented from an insufflator, nebulizer press pack or other convenient means of administering an aerosol spray. The pressurized pack may contain a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be dosed by providing a valve. Alternatively, for administration by inhalation or insufflation, the pharmaceutical formulations may take the form of a dry powder composition, for example a powder mix of the compound and a suitable powder base such as lactose or starch. The powder composition may be in unit dosage form, for example as a capsule, cartridge (cartridge), gelatin or blister tablet from which the powder may be administered by means of an inhaler or insufflator.

It will be understood that the compounds and compositions described herein may contain, in addition to the ingredients specifically mentioned above, other agents conventional in the art having regard to the type of formulation in question, for example, ingredients suitable for oral administration may include flavouring agents.

Preparation

It should be noted that any of the compositions and compounds described herein can be used in any of the formulations discussed in this section, which are not intended to be limiting and should not be construed as limiting.

The compounds or compositions described herein can be administered as vesicles, such as Liposomes (see, e.g., Langer, Science 1990,249, 1527) -1533; Treat et al, lipids in the Therapy of Infectious diseases and Cancer, Lopez-Bernstein and Fidler, Ed., Liss, N.Y., pp.353-365,1989). The compounds and pharmaceutical compositions described herein may also be administered in a controlled release system. In one embodiment, pumps may be used (see, Sefton,1987, CRC crit, Ref. biomed. Eng.14: 201; Buchwald et al, Surgery, 198088,507; Saudek et al, N.Engl. J. Med.1989,321, (574). furthermore, Controlled Release systems may be placed in the vicinity of the target of treatment (see, Goodson, Medical Applications of Controlled Release,1984, Vol.2, pp.115-138.) the pharmaceutical compositions described herein may also contain the active ingredients in a form suitable for oral use, for example, in the form of tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs A non-toxic pharmaceutically acceptable excipient. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents; fillers such as microcrystalline cellulose, silicified microcrystalline cellulose, pregelatinized starch, lactose, calcium hydrogen phosphate or compressible sugars; binders such as hypromellose, povidone, or starch paste; disintegrants such as croscarmellose sodium, crospovidone or sodium starch glycolate; surfactants such as sodium lauryl sulfate, and/or lubricants, and processing aids such as talc, croscarmellose sodium, corn starch, or alginic acid; binding agents (bindingbinders) such as starch, gelatin, polyvinylpyrrolidone or gum arabic, and lubricating agents such as magnesium stearate, stearic acid or colloidal silicon dioxide, and optionally talc. The tablets may be uncoated or coated by known methods in order to mask the taste of the drug or to delay disintegration and absorption in the gastrointestinal tract and thereby provide a longer lasting effect. For example, a water soluble taste masking material such as hydroxypropylmethyl cellulose or hydroxypropyl cellulose, or a time delay material such as ethyl cellulose or cellulose acetate butyrate may be suitably used. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier, for example polyethylene glycol, or an oily medium such as peanut oil, liquid paraffin or olive oil. The capsule and tablet dosage forms may be prepared by a variety of processing techniques, including dry mixing and wet granulation techniques. In the dry-blend manufacturing process, the drug may be included in the dosage form by dry-blending with excipients and then encapsulating into a capsule shell or compressing into tablet form. The dry mixing operation may be performed stepwise and includes a screening step between mixing steps to facilitate the formation of a homogeneous mixture. In the wet granulation manufacturing process, the drug may be added to the dry excipients, mixed, and then the binder solution added, or the drug may be dissolved and added as a solution in the granulation section. In the wet granulation technique, if a surfactant is used, it may be added to the dry excipient, or it may be added to the binder solution and contained therein in the form of a solution. Capsule dosage forms may also be prepared by dissolving the drug in a material that can be filled into a hard gelatin capsule shell and is compatible with a hard gelatin capsule shell that can then be banded and sealed. Capsule and tablet dosage forms may also be prepared by dissolving the drug in a molten form of a material such as high molecular weight polyethylene glycol, cooling to a solid form, grinding, and incorporating this material into conventional capsule and tablet processing.

Aqueous suspensions contain the active ingredient in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecavinyl-oxycetanol (heptadeca-ethyleneoxy-ethylene glycol), or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (e.g., ethyl or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents, and one or more sweetening agents (e.g., sucrose, saccharin, or aspartame).

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions can be preserved by the addition of antioxidants, such as butylated hydroxyanisole or alpha-tocopherol.

The active ingredient is formed by the addition of water to dispersible powders and granules suitable for the preparation of an aqueous suspension in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Other excipients, for example sweetening, flavouring and colouring agents, may also be present. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical composition may also be in the form of an oil-in-water emulsion. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin, or a mixture of these. Suitable emulsifying agents may be naturally-occurring phosphatides (e.g. soy bean lecithin), and esters or partial esters derived from fatty acids and hexitol anhydrides (e.g. sorbitan monooleate), and condensation products of the said partial esters with ethylene oxide (e.g. polyoxyethylene sorbitan monooleate). The emulsions may also contain sweetening, flavoring, preservative and antioxidant agents.

Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a flavouring and colouring agent and an antioxidant.

The pharmaceutical compositions may be in the form of sterile injectable aqueous solutions. Among the acceptable carriers and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may also be sterile injectableA injectable oil-in-water microemulsion wherein the active ingredient is dissolved in the oil phase. For example, the active ingredient may first be dissolved in a mixture of soybean oil or soybean lecithin. The oily solution is then introduced into a mixture of water and glycerol and processed to form a microemulsion. The injectable solutions or microemulsions may be introduced into the bloodstream of a patient by local bolus injection. Alternatively, it may be advantageous to administer the solution or microemulsion in such a way as to maintain a constant circulating concentration of the instant compound. To maintain such a constant concentration, a continuous intravenous administration set may be used. An example of such a device is DeltecCADD-PLUS^TMModel 5400 intravenous pump. The pharmaceutical compositions may be in the form of sterile injectable aqueous or oleaginous suspensions for intramuscular and subcutaneous administration. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The pharmaceutical compositions may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the inhibitor with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, glycerogelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights, and fatty acid esters of polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds or compositions of the present invention are used for topical administration. Topical application as used herein may include mouth washes and gargles.

The pharmaceutical compositions may be administered in intranasal form by topical use of suitable intranasal vehicles and delivery devices, or by transdermal routes, using those transdermal patch forms well known to those skilled in the art.

The formulations may conveniently be presented in unit dosage form and may be prepared by methods well known in the art of pharmacy. All methods include the step of bringing into association a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug or solvate thereof ("active ingredient") with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired formulation. Methods for preparing various pharmaceutical compositions using specific amounts of active compounds are known or will be apparent to those skilled in the art. For administration in transdermal form, the dosage form will, of course, be continuous rather than intermittent throughout the dosage regimen.

Dosage form

The amount of the pharmaceutical composition to be administered depends, inter alia, on the mammal to be treated. In the case of administration of a pharmaceutical composition to a human subject, the daily dosage will generally be determined by the prescribing physician, with the dosage generally being a function of the age, sex, diet, body weight, general health and response of the individual patient, the severity of the patient's symptoms, the precise indication or condition being treated, the severity of the indication or condition being treated, the time of administration, the route of administration, the disposition of the composition, the rate of excretion, the drug combination, and the discretion of the prescribing physician. Moreover, the route of administration may vary depending on the condition and its severity. The pharmaceutical composition may be in unit dosage form. In such form, the formulation is subdivided into unit doses containing appropriate quantities of the active ingredient, e.g., an effective amount to achieve the intended purpose. It is within the skill of the art to determine the appropriate dosage for a particular situation. In general, treatment is initiated with smaller doses than the optimal dose of the compound. Thereafter, with a small increaseThe dosage is such that the optimum effect under the circumstances is achieved. For convenience, the total daily dose may be divided and administered in portions during the day, if desired. The amount and frequency of administration of the compounds described herein, and if appropriate, of other therapeutic agents and/or therapies, is adjusted at the discretion of the attending clinician (physician) taking into account the factors described above. Thus, the dosage of pharmaceutical compositions administered may vary widely. The amount administered may be from about 0.001mg/kg body weight to about 100mg/kg body weight per day (administered in a single dose or in divided doses), or at least about 0.1mg/kg body weight per day. Specific therapeutic doses may include, for example, from about 0.01mg to about 7000mg of the compound, or, for example, from about 0.05mg to about 2500 mg. The amount of active compound in a unit dosage formulation may be varied or adjusted within the range of about 0.1mg to 1000mg, about 1mg to 300mg, or 10mg to 200mg, depending on the particular use. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases even larger doses may be employed without causing any harmful side effects, e.g., by dividing such larger doses into several small doses administered throughout the day. The amount administered will depend on the particular IC of the compound used ₅₀The value changes. In combination administration where the compound is not the only therapy, it is possible to administer lower amounts of the compound and still have a therapeutic or prophylactic effect.

Other dosages are provided throughout the specification and claims.

Dosage forms

The pharmaceutical compositions may be in the form of, for example, tablets, capsules, pills, powders, sustained release formulations, solutions, suspensions suitable for oral administration, sterile solutions, suspensions or emulsions suitable for parenteral injection, ointments or creams suitable for topical administration, or suppositories suitable for rectal administration. The pharmaceutical compositions may be in unit dosage form suitable for single administration of the correct dosage. The pharmaceutical compositions will include conventional pharmaceutical carriers or excipients and the compounds of the present invention as active ingredients. In addition, it may include other pharmaceutically or pharmacologically acceptable agents, carriers, adjuvants and the like.

Exemplary parenteral administration forms include solutions or suspensions of the active compounds in sterile aqueous solutions (e.g., aqueous propylene glycol or glucose solutions). Such dosage forms may be suitably buffered if desired.

Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. The pharmaceutical composition may, if desired, contain other ingredients such as flavouring agents, binders, excipients and the like. Thus, for oral administration, tablets contain various excipients, for example citric acid, together with various disintegrants (e.g. starch, alginic acid and certain complex silicates) and binding agents (e.g. sucrose, gelatin and acacia). Additionally, lubricants such as magnesium stearate, sodium lauryl sulfate and talc are often used for tableting purposes. Solid compositions of a similar type may also be used in soft-filled gelatin capsules and hard-filled gelatin capsules comprising lactose and a high molecular weight polyethylene glycol. When aqueous suspensions or elixirs are desired for oral administration, the active compound therein may be combined with various sweetening or flavoring agents, coloring matter or dyes, and, if desired, emulsifying or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin or combinations thereof.

Methods of formulating various pharmaceutical compositions containing a particular amount of an active compound are known, or will be apparent, to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, ester, Pa., 18 th edition (1990).

Combination therapy

The compounds described herein, or pharmaceutically acceptable salts, solvates, polymorphs, esters, amides, tautomers, prodrugs, hydrates or derivatives thereof, can be administered as a monotherapy. A compound described herein, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, can also be administered in combination with one or more other therapies.

Also described herein is N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form a), which can be administered as monotherapy. Crystalline polymorph form a of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide can also be administered in combination with one or more other therapies.

By way of example only, if one of the side effects experienced by a patient when receiving one of the compounds described herein is hypertension, then an antihypertensive agent and the compound may be suitably administered in combination. Alternatively, by way of example only, the therapeutic benefit of one of the compounds described herein may be enhanced by administration of an adjuvant (i.e., the adjuvant itself may have only minimal therapeutic benefit, but in combination with another therapeutic agent, the overall therapeutic benefit to the patient is increased). Alternatively, by way of example only, the benefit experienced by a patient may be enhanced by administering one of the compounds described herein in combination with another therapeutic agent that also has therapeutic benefit (which also includes a treatment regimen). By way of example only, in the treatment of diabetes involving administration of one of the compounds described herein, the therapeutic benefit may also be enhanced by providing the patient with another therapeutic agent for diabetes. In any case, regardless of the disease, disorder, or condition being treated, the overall benefit experienced by the patient may be a simple addition of the two therapeutic agents, or the patient may experience a synergistic benefit.

Other therapies include, but are not limited to, administration of other therapeutic agents, radiation therapy, or both. Where the compounds described herein are administered in combination with other therapeutic agents, the compounds described herein need not be administered in the same pharmaceutical composition as the other therapeutic agents and may be administered by different routes due to different physical and chemical characteristics. For example, the compound/composition may be administered orally to produce and maintain good blood levels thereof, while another therapeutic agent may be administered intravenously. In the possibility ofIn the case of (a), determining the mode of administration and suitability for administration in the same pharmaceutical composition is well known to the skilled clinician. Initial administration can be carried out according to protocols established in the art, and then, depending on the observed effect, a skilled clinician can adjust the dosage, mode of administration, and timing of administration. The particular choice of compound (and, where appropriate, other therapeutic agents and/or radiation) will depend upon the diagnosis of the attending physician and their judgment as to the condition of the patient and the appropriate treatment regimen. Other therapeutic agents may include chemotherapeutic agents such as anti-neoplastic substances, for example selected from mitotic inhibitors (e.g. vinblastine); alkylating agents (e.g., cisplatin, carboplatin, and cyclophosphamide); antimetabolites (e.g., 5-fluorouracil, cytarabine and hydroxyurea, or antimetabolites such as N- (5- [ N- (3, 4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)) -N-methylamino), as disclosed, for example, in European patent application No. 239362 ]-2-thenoyl) -L-glutamic acid); a growth factor inhibitor; a cell cycle inhibitor; intercalating antibiotics (e.g., doxorubicin and bleomycin); enzymes (e.g., interferons); and anti-hormones (e.g. anti-estrogens such as Nolvadex)^TM(tamoxifen) or, for example, antiandrogens such as Casodex^TM(4 '-cyano-3- (4-fluorophenylsulfonyl) -2-hydroxy-2-methyl-3' - (trifluoromethyl) propionylaniline)). Such combination therapy may be achieved by administering the therapeutic components simultaneously, sequentially or separately.

The compounds and compositions described herein (and the appropriate chemotherapeutic agent and/or radiation therapy) may be administered together (e.g., simultaneously, substantially simultaneously, or within the same treatment regimen), or sequentially, depending on the nature of the disease or condition of the patient, and the actual choice of chemotherapeutic agent and/or radiation therapy to be used in combination with the compound/composition (i.e., within a single treatment regimen).

In combined administration and use, the compound/composition and chemotherapeutic agent and/or radiation therapy need not be administered simultaneously or substantially simultaneously, and the initial order of administration of the compound/composition and chemotherapeutic agent and/or radiation therapy may not be important. Thus, the compounds/compositions of the present invention may be administered first, followed by administration of chemotherapeutic agents and/or radiation therapy; alternatively, the chemotherapeutic agent and/or radiation therapy may be administered first, followed by administration of the compounds/compositions of the present invention. This alternative method of administration may be repeated during a single treatment regimen. After evaluating a patient for a disease and condition to be treated, it is within the knowledge of a skilled physician to determine the order of administration and the number of times each therapy is to be repeated during a treatment regimen. For example, the chemotherapeutic agent and/or radiation therapy may be administered first, particularly if it is a cytotoxic agent, and then treatment is continued by subsequently administering the compounds/compositions of the invention, followed by administration of the chemotherapeutic agent and/or radiation, etc., where deemed advantageous, until the treatment regimen is over. Thus, based on experience and knowledge, as treatment progresses, the practitioner may adjust each dosing regimen of the compound/composition for treatment according to the needs of the individual patient. In determining whether a treatment at an administered dose is effective, the attending clinician will consider the overall health of the patient, as well as more specific signs, such as alleviation of disease-related symptoms, inhibition of tumor growth, actual shrinkage of the tumor, or inhibition of metastasis. The size of the tumor can be measured by standard methods such as radiological studies, e.g. CAT or MRI scans, and subsequent measurements can be used to determine whether the growth of the tumor has been inhibited or even reversed. Relief of disease-related symptoms such as pain and improvement of the overall condition may also be used to help judge the effectiveness of the treatment.

Specific non-limiting examples of possible combination therapies include the combination of a compound of the present invention with agents in the following drug treatment categories as described below. These lists should not be construed as closed but should be used as illustrative examples common to the relevant therapeutic arts at present. Furthermore, the combination regimen may include various routes of administration, and should include oral, intravenous, intraocular, subcutaneous, topical dermal and topical inhalation administration.

For the treatment of neoplastic diseases, proliferative disorders and cancer, the compounds of the invention may be administered in combination with an agent selected from the group consisting of: aromatase inhibitors, antiestrogens, antiandrogens, corticosteroids, gonadorelin agonists, topoisomerase 1 and 2 inhibitors, microtubule activators, alkylating agents, nitrosoureas, antineoplastic antimetabolites, platinum-containing compounds, lipid or protein kinase targeting agents, IMiDs, protein or lipid phosphatase targeting agents, antiangiogenic agents, Akt inhibitors, IGF-I inhibitors, FGF3 modulators, mTOR inhibitors, Smac mimetics, HDAC inhibitors, agents that induce cell differentiation, bradykinin 1 receptor antagonists, angiotensin II antagonists, cyclooxygenase inhibitors, heparanase (heparanase) inhibitors, lymphokine inhibitors, cytokine inhibitors, IKK inhibitors, P38MAPK inhibitors, ARRY-797, HSP90 inhibitors, multikinase inhibitors, bisphosphonates (bisphosphanate), rapamycin derivatives, anti-apoptotic pathway inhibitors, apoptotic pathway agonists, apoptosis pathway inhibitors, inhibitors of the activity of an enzyme, and/or of the activity of an enzyme, PPAR agonists, RAR agonists, Ras isoform inhibitors, telomerase inhibitors, protease inhibitors, metalloproteinase inhibitors, aminopeptidase inhibitors, SHIP activator-AQX-MN 100, Humax-CD20(ofatumumab), CD20 antagonists, IL 2-diphtheria toxin fusions.

For the treatment of neoplastic diseases, proliferative disorders and cancer, the compounds of the invention may be administered in combination with an agent selected from the group consisting of: dacarbazine (DTIC), actinomycin C₂Actinomycin C₃Actinomycin D and actinomycin F₁Cyclophosphamide, melphalan, estramustine, maytansinol, rifamycin, varicocele, doxorubicin, daunorubicin, epirubicin, idarubicin, ditobicin, nordaunorubicin, idarubicin, epirubicin, isorubicin, mitoxantrone, bleomycin A₂And bleomycin B, camptothecin, irinotecan, RTM, topotecan, RTM, 9-aminocamptothecin, 10, 11-methylenedioxycamptothecin, 9-nitrocamptothecin, bortezomib, temozolomide, TAS103, NPI0052, combretastatin A-2, combretastatin A-4, calicheamicin, neocarzinostatin, epothilone A, epothilone B, epothilone C and semisynthetic variants, sahetin, RTM, merozoite, RTM, CD40 antibodies, asparaginase, leukapheresis, RTMInterferon, leuprolide and pemphigenise, 5-fluorouracil, fluorodeoxyuridine, ptorafur, 5' -deoxyfluorouridine, UFT, MITC, S-1 capecitabine, diethylstilbestrol, tamoxifen, toremifene, tolmetrexed (tolmoudex), thymititaq, flutamide, fluomethyltestosterone, bicalutamide, finasteride, estradiol, trovaxifene, dexamethasone, leuprolide acetate, estramustine, droloxifene, medroxyprogesterone, megestrol acetate, aminoglutethimide, testolactone, testosterone, diethylstilbestrol, hydroxyprogesterone, mitomycin A, mitomycin B and mitomycin C, pofelycin, cisplatin, carboplatin, oxaliplatin, tetraplatin, platinum-DACH, ormaplatin, thalidomide, lenalidomide, CI-973, Rad, CHIR258, telithromycin, SAHA, saaag 216, Turafludoxylamine, Turafluoride, Tubifeprinolide, JM-17, and, Podophyllotoxin, epipodophyllotoxin, etoposide, teniposide, tassel, rtm, iressa, rtm, imatinib, rtm, miltefosine, rtm, perifosin, rtm, aminopterin, methotrexate, 6-mercaptopurine, thioguanine, azatuoprine, allopurinol, cladribine, fludarabine, pentostatin, 2-chloroadenosine, deoxycytidine, cytarabine, azacitidine, 5-azacytosine, gemcitabine, 5-azacytosine-arabinoside, vincristine, vinorelbine epoxide, isocartrine and vindesine, paclitaxel, taxotere and docetaxel.

For the treatment of inflammatory diseases or pain, the compounds of the present invention and pharmaceutically acceptable salts of the compounds may be administered in combination with an agent selected from the group consisting of: corticosteroids, non-steroidal anti-inflammatory drugs, muscle relaxants and combinations thereof with other agents, anesthetics and combinations thereof with other agents, expectorants and combinations thereof with other agents, antidepressants, anticonvulsants and combinations thereof; antihypertensives, opioids, topical cannabinoids, capsaicin, betamethasone dipropionate (synergistic and non-synergistic), betamethasone valerate, clobetasol propionate, prednisone, methylprednisolone, diflorasone diacetate, halobetasol propionate, amcinonide, dexamethasone, desoximetasone (desoxsimethone), fluocinonide, fluocinolone acetonide, halcinonide (halocinonide), clocortolone pivalate, desoximetasone (desoximetasone), flurandrenolone acetonide, salicylate, ibuprofen, ketoprofen, etodolac, diclofenac, meclofenamate sodium, naproxen, piroxicam, celecoxib, cyclobenzaprine, baclofen, cyclobenzaprine/lidocaine, baclofen/cyclobenzaprine, cyclobenzaprine/ketoprofen, lidocaine/D-glucose, Prilocaine, EMLA cream (eutectic mixture of local anesthetics (lidocaine 2.5% and prilocaine 2.5%), guaifenesin/ketoprofen/cyclobenzaprine, amitriptyline (amitriptiline), doxepin, desipramine, imipramine, amoxapine, clomipramine, nortriptyline, protriptyline, duloxetine, mirtazapine (mirtazepine), nisoxetine, maprotiline, reboxetine, fluoxetine, fluvoxamine, carbamazepine, fenoxaprop-e, lamotrigine, topiramate, tiagabine, oxcarbazepine (carbamazepine), zonisamide, mexiletine, gabapentin/clonidine, gabapentin/carbamazepine, carbamazepine/cycloxapriapine, antihypertensive including benzalkonium, codeine, loratadine, loperamide, trospidone, ketoprofen, doxoramide, doxorafen, doxorapride, doxoraprine, doxorapridine, doxoraprine, Levorphanol, butorphanol, menthol, wintergreen oil, camphor, eucalyptus oil, and turpentine oil; CB1/CB2 ligand, acetaminophen, infliximab, a nitric oxide synthase inhibitor, in particular an inducible inhibitor of nitric oxide synthase, a PDE4 inhibitor-mechanism similar to that of ibudilast (AV-411), CDC-801, a JNK inhibitor-CC-401, a TNF/PDE4 combination inhibitor-CDC-998, an IL1 antagonist such as anakinra-Kineret, AMG 108, a (mAb) targeting IL-1, a SHIP activator-AQX-MN 100, a C5 antagonist, a C5a inhibitor, Pekezumab, a pyrimidine synthesis inhibitor, a lymphokine inhibitor, a cytokine inhibitor, an IKK inhibitor, a P38MAPK inhibitor, HSP AR797, a HSP90 inhibitor, a multi-kinase inhibitor, a bisphosphonate, a PPAR agonist, a Cox1 and Cox2 inhibitor, an anti-CD 4 therapy, a B-cell inhibitor, a COX/LOX dual inhibitor, Immunosuppressive drugs, iNOS inhibitors, NSAIDs, sPLA2 inhibitors, colchicine, allopurinol, oxypurinol, gold, reed-auranofin, febuxostat, Puricase, PEG-uricase preparations, benzbromarone, long-acting beta-2 agonists (LABA), salmeterol (SereventDiskus) and formoterol (fordil), leukotriene modulators including montelukast (Singulair) and zafirlukast (accelate), inhaled cromoglycic acid (oral) or nedocromil (balade), theophylline. Short-acting beta-2 agonists, ipratropium (Atrovent), immunotherapeutics- (allergic desensitization injections), anti-IgE monoclonal antibody-Xolair, common DMARDs include: hydroxychloroquine (plaquinil), the gold compounds auranofin (regained), sulfasalazine (Azulfidine), minocycline (Dynacin, Minocin) and methotrexate (rhematrex), leflunomide (Arava), azathioprine (magnolia), cyclosporine (Neoral, Sandimmune) and cyclophosphamide (Cytoxan), antibiotics, CD80 antagonists, co-stimulatory factor antagonists, Humax-CD20 (ofatumumab); CD20 antagonists, MEK inhibitors, NF kappa-B inhibitors, anti-B-cell antibodies, disitumumab, mAbs that specifically target receptor activators of nuclear factor kappa B ligand (RANKL). IL17 inactivating antibodies, IL-17 receptor antagonists/inhibitors, CTLA inhibitors, CD20 inhibitors, soluble VEGFR-1 receptors, anti-VEGFR-1 receptor antibodies, anti-VEGF antibodies, integrin receptor antagonists, selectin inhibitors, P-selectin inhibitors and E-selectin inhibitors, phospholipase A2 inhibitors, lipoxygenase inhibitors, RANKL and RANK antagonists/antibodies, osteoprotegerin antagonists, lymphotoxin inhibitors, B-lymphocyte stimulators, MCP-1 inhibitors, MIF inhibitors, CD2 inhibitors, CD3 inhibitors, CD4 inhibitors, CD25 inhibitors, CD40 inhibitors and CD40 ligand CD152(CTLA4) inhibitors, macrolide immunosuppressants, selective inhibitors of nucleotide metabolism, inhibitors of chemotaxis, CXC receptor and CXC ligand inhibitors, Chemokine antagonists, leukocyte chemotaxis inhibitors, adhesion molecule blockers, selectin lymphocyte function antigen-1 (LFA-1, CD11a) antagonists, very late antigen-4 (VLA-4) antagonists, matrix metalloproteinase inhibitors, elastase inhibitors, cathepsin inhibitors.

For the treatment of ocular disorders and diseases, the compounds of the present invention and pharmaceutically acceptable salts of said compounds may be administered with an agent selected from the group consisting of: beta-blockers, carbonic anhydrase inhibitors, alpha-adrenergic antagonists, and beta-adrenergic antagonists include alpha l-adrenergic antagonists, alpha 2 agonists, miotics, prostaglandin analogs, corticosteroids, and immunosuppressive drugs.

For the treatment of ocular disorders and diseases, the compounds of the present invention and pharmaceutically acceptable salts of said compounds may be administered in combination with an agent selected from the group consisting of: timolol, betaxolol, levobetaxolol, carteolol, levobunolol, propranolol, brinzolamide, dorzolamide, niprolide, acerolane, brimonidine, pilocarpine, epinephrine, latanoprost, travoprost, bimatoprost, unoprostone, dexamethasone, prednisone, methylprednisolone, azathioprine, cyclosporine, and immunoglobulins.

For the treatment of autoimmune diseases, the compounds of the invention or pharmaceutically acceptable salts of the compounds may be administered in combination with an agent selected from the group consisting of: corticosteroids, immunosuppressive drugs, prostaglandin analogs, and antimetabolites.

For the treatment of autoimmune diseases, the compounds of the invention may be administered in combination with an agent selected from the group consisting of: dexamethasone, prednisone, methylprednisolone, azathioprine, cyclosporine, immunoglobulin, latanoprost, travoprost, bimatoprost, unoprostone, infliximab, rituximab, methotrexate, non-steroidal anti-inflammatory drugs, muscle relaxants and combinations thereof with other agents, anesthetics and combinations thereof with other agents, expectorants and combinations thereof with other agents, antidepressants, anticonvulsants and combinations thereof; antihypertensives, opioids, topical cannabinoids, and other agents, such as capsaicin, betamethasone dipropionate (both potentiated and non-potentiated), betamethasone valerate, clobetasol propionate, prednisone, methylprednisolone, diflunisal diacetate, halobetasol propionate, amcinonide, dexamethasone, desoximetasone, fluocinolone acetonide, fluocinonide, halcinonide, clocortolone pivalate, desoximetasone, fluroxypyr acetonide, salicylate, ibuprofen, ketoprofen, etodolac, diclofenac, meclofenamate sodium, naproxen, piroxicam, celecoxib, cyclobenzaprine, baclofen, cyclobenzaprine/lidocaine, baclofen/cyclobenzaprine, cyclobenzaprine/lidocaine/ketoprofen, lidocaine/deoxy-D-glucose, Prilocaine, EMLA cream (eutectic mixture of local anesthetics (lidocaine 2.5% and prilocaine 2.5%), guaifenesin/ketoprofen/cyclobenzaprine, amitriptyline, doxepin, desipramine, imipramine, amoxapine, clomipramine, nortriptyline, protriptyline, duloxetine, mirtazapine, nisoxetine, maprotiline, reboxetine, fluoxetine, fluvoxamine, carbamazepine, felbamine, lamotrigine, topiramate, tiagabine, oxcarbazepine, carbamazepine, zonisamide, mexiletine, gabapentin/codine, gabapentin/carbamazepine, carbamazepine/cyclobenzaprine, antihypertensives including clonidine, codeine, loperamide, tramadol, morphine, fentanyl, oxycodone, hydrocodone, levorphanol, pramipexorphanol, imipramipeline, pramipexole, doxine, clozapine, cloxacine, Menthol, wintergreen oil, camphor, eucalyptus oil, turpentine oil; CB1/CB2 ligand, acetaminophen, infliximab; nitric oxide synthase inhibitors, in particular inducible nitric oxide synthase inhibitors; and other agents, such as capsaicin. PDE4 inhibitors-similar in mechanism to ibudilast (AV-411), CDC-801, JNK inhibitors-CC-401, TNF/PDE4 combination inhibitors-CDC-998, IL1 antagonists such as anakinra-Kineret, AMG 108, IL-1 targeting (mAb), SHIP activators-AQX-MN 100, C5 antagonists, C5a inhibitors, pekelizumab, pyrimidine synthesis inhibitors, lymphokine inhibitors, cytokine inhibitors, IKK inhibitors, P38MAPK inhibitors, ARRY-797, HSP90 inhibitors, multi-kinase inhibitors, bisphosphonates, PPAR agonists, Cox1 and Cox2 inhibitors, anti-CD 4 therapy, B-cell inhibitors, COX/LOX dual inhibitors, immunosuppressive drugs, iNOS inhibitors, NSAIDs, sPLA2 inhibitors, colchicine, allopurinol, oxypurinol, gold, Rede-auranofin, febuxostat, Puricase, PEG-uricase preparations, benzbromarone, long-acting beta-2 agonists (LABA), salmeterol (Serevent Diskus) and formoterol (formadil), leukotriene modulators including montelukast (Singulair) and zafirlukast (Acclate). Inhaled cromolyn (oral) or nedocromil (Tilade), theophylline. Short-acting beta-2 agonists, ipratropium (Atrovent), immunotherapeutics- (allergic desensitization injection), anti-IgE monoclonal antibody-Xolair, common DMARDs including hydroxychloroquine (Plaquenil), the gold compound auranofin (reed), sulfasalazine (Azulfidine), minocycline (Dynacin, Minocin) and methotrexate (Rheumatrex), leflunomide (Arava), azathioprine (imulan), cyclosporine (Neoral, sandimune) and cyclophosphamide (Cytoxan), antibiotics, CD80 antagonists, co-stimulatory factor antagonists, Humax-CD20 (ofatumumab); CD20 antagonists, MEK inhibitors, NF κ B inhibitors, anti-B cell antibodies, disitumumab, mabs that specifically target receptor activators of nuclear factor κ B ligand (RANKL). IL17 inactivating antibodies, IL-17 receptor antagonists/inhibitors, CTLA inhibitors, CD20 inhibitors, soluble VEGFR-1 receptors, anti-VEGFR-1 receptor antibodies, anti-VEGF antibodies, integrin receptor antagonists, selectin inhibitors, P-selectin inhibitors and E-selectin inhibitors, phospholipase A2 inhibitors, lipoxygenase inhibitors, RANKL and RANK antagonists/antibodies, osteoprotegerin antagonists, lymphotoxin inhibitors, B-lymphocyte stimulators, MCP-1 inhibitors, MIF inhibitors, CD2 inhibitors, CD3 inhibitors, CD4 inhibitors, CD25 inhibitors, CD40 inhibitors and CD40 ligand CD152(CTLA4) inhibitors, macrolide immunosuppressants, selective inhibitors of nucleotide metabolism, inhibitors of chemotaxis, CXC receptor and CXC ligand inhibitors, Chemokine antagonists, leukocyte chemotaxis inhibitors, adhesion molecule blockers, selectin lymphocyte function antigen-1 (LFA-1, CD11a) antagonists, very late antigen-4 (VLA-4) antagonists, matrix metalloproteinase inhibitors, elastase inhibitors, cathepsin inhibitors.

For the treatment of metabolic disorders, the compounds of the present invention and pharmaceutically acceptable salts of said compounds may be administered in combination with an agent selected from the group consisting of: insulin, insulin derivatives and mimetics, insulin secretagogues, insulin sensitizers, biguanide agents, alpha-glucosidase inhibitors, insulinotropic sulfonylurea receptor ligands, protein tyrosine phosphatase-1B (PTP-1B) inhibitors, GSK3 (glycogen synthase kinase-3) inhibitors, GLP-1 (glucagon-like peptide-1), GLP-1 analogs, DPPIV (dipeptidyl peptidase IV) inhibitors, RXR ligand sodium-dependent glucose cotransporter inhibitors, glycogen phosphorylase A inhibitors, AGE disruptors, PPAR modulators, LXR and FXR modulators, non-glitazone PPARS agonists, selective glucocorticoid antagonists, metformin, glipizide, glyburide, submenuide, meglitinide, nateglinide, repaglinide, PT-112, SB-517955, SB4195052, SB-216763, NN-57-05441, NN-57-05445, GW-0791, AGN-. sup.194.sup.204, T-1095, BAY R3401, acarbose Exendin-4, DPP728, LAF237, vildagliptin, MK-0431, saxagliptin, GSK23A, pioglitazone, rosiglitazone, (R) -1- {4- [ 5-methyl-2- (4-trifluoromethyl-phenyl) -oxazol-4-ylmethoxy ] -benzenesulfonyl }2, 3-dihydro-1H-indole-2-carboxylic acid described in patent application WO 03/043985, Compound 19 as in example 4, and GI-262570.

Disease and disorder

Described herein are methods of treating a disease in a subject suffering from the disease, comprising administering to the subject an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof.

Also described herein are methods of treating a disease or disorder in a subject suffering from the disease or disorder, comprising administering to the subject an effective amount of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form a). The present invention relates to the use of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form a) for the preparation of a medicament for the treatment of a disease or disorder.

In some embodiments, the invention relates to the prevention or treatment of any disease or disorder in which MEK kinase plays a role, including, without limitation: neoplastic diseases, hematologic diseases, inflammatory diseases, ocular diseases, neurological diseases, immunological diseases, cardiovascular diseases, and dermatological diseases, as well as diseases caused by excessive or unregulated production of pro-inflammatory cytokines, including, for example, excessive or unregulated production of TNF, IL-1, IL-6, and IL-8 in humans or other mammals. The present invention relates to such use of the compounds, and to the use of the compounds in the manufacture of a medicament for the treatment of such cytokine mediated diseases or conditions. In addition, the invention relates to the administration of an effective amount of a MEK inhibitor to a human to treat any such disease or disorder.

Diseases or disorders in which MEK kinases act directly or via pro-inflammatory cytokines (including cytokines TNF, IL-1, IL-6, and IL-8) include, without limitation: dry eye, glaucoma, autoimmune diseases, inflammatory diseases, destructive bone diseases, proliferative diseases, neurodegenerative disorders, viral diseases, allergies, infectious diseases, heart attacks, angiogenic disorders, reperfusion/ischemia in the stroke, vascular hyperplasia, organ hypoxia, cardiac hypertrophy, platelet aggregation by thrombin, and conditions associated with prostaglandin endoperoxidase synthase-2 (COX-2).

In certain aspects of the invention, the disease is a hyperproliferative condition of the human or animal body, including, but not limited to, cancer, hyperplasia, restenosis, inflammation, immune diseases, cardiac hypertrophy, atherosclerosis, pain, migraine, angiogenesis-related conditions or disorders, proliferation caused following medical conditions (including, but not limited to, surgery, angioplasty, or other conditions).

In a further embodiment, the hyperproliferative condition is selected from the group consisting of a hematologic disease cancer and a non-hematologic disease cancer. In still other embodiments, the hematologic disease cancer is selected from multiple myeloma, leukemia, and lymphoma. In a still further embodiment, the leukemia is selected from acute leukemia and chronic leukemia. In still further embodiments, the acute leukemia is selected from Acute Lymphocytic Leukemia (ALL) and acute non-lymphocytic leukemia (ANLL). In still further embodiments, the chronic leukemia is selected from Chronic Lymphocytic Leukemia (CLL) and Chronic Myelogenous Leukemia (CML). In further embodiments, the lymphoma is selected from hodgkin's lymphoma and non-hodgkin's lymphoma. In a further embodiment, the hematological disease cancer is multiple myeloma. In other embodiments, the hematologic disease cancer is low grade cancer, moderate cancer, or high grade cancer. In other embodiments, the non-hematologic disease cancer is selected from brain cancer, head and neck cancer, lung cancer, breast cancer, cancer of the reproductive system, cancer of the digestive system, pancreatic cancer, and cancer of the urinary system. In a further embodiment, the cancer of the digestive system is a cancer of the upper digestive tract or colorectal cancer. In further embodiments, the cancer of the urinary system is bladder cancer or renal cell carcinoma. In a further embodiment, the cancer of the reproductive system is prostate cancer.

Other types of cancers that may be treated using the compounds and methods described herein include: cancers of the oral cavity and pharynx, cancers of the respiratory system, cancers of the bones and joints, cancers of soft tissues, skin cancers, cancers of the reproductive system, cancers of the eyes and orbit, cancers of the nervous system, cancers of the lymphatic system, and cancers of the endocrine system. In certain embodiments, these cancers may be selected from the group consisting of cancers of the tongue, mouth, pharynx, or other oral cavity; esophageal cancer, gastric cancer, or small bowel cancer; colon or rectal cancer, anal cancer, or anorectal cancer; liver cancer, intrahepatic bile duct cancer, gallbladder cancer, pancreatic cancer, or other cancers of the gallbladder or digestive organs; laryngeal, bronchial and other respiratory cancers; heart cancer, melanoma, basal cell carcinoma, squamous cell carcinoma, other non-epithelial cancers; uterine cancer or cervical cancer; uterine body cancer; ovarian, vulvar, vaginal, or other cancers of the female genitalia; prostate cancer, testicular cancer, cancer of the penis or other male genitalia; urinary bladder cancer; cancer of the kidney; kidney, pelvis, or urinary tract cancer, or other cancers of the genitourinary organs; thyroid cancer or other cancer of the endocrine glands; chronic lymphocytic leukemia; and granulocytic and unicellular cutaneous T-cell lymphoma.

Still other types of cancers that may be treated using the compounds and methods described herein include: adenocarcinoma, angiosarcoma, astrocytoma, acoustic neuroma, glioblastoma multiforme, basal cell carcinoma, glioblastoma multiforme (blastoglioma), chondrosarcoma, choriocarcinoma, chordoma, craniopharyngioma, cutaneous melanoma, cystadenocarcinoma, endotheliosarcoma, embryonal carcinoma, ependymoma, ewing's tumor, epithelial carcinoma, fibrosarcoma, gastric carcinoma, genitourinary tract cancer, glioblastoma multiforme, hemangioblastoma, hepatocellular carcinoma, liver carcinoma, kaposi's sarcoma, large cell carcinoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, medullary carcinoma of the thyroid gland, medulloblastoma, meningioma, mesothelioma, myeloma, myxosarcoma, neuroblastoma, neurofibrosarcoma, oligodendroglioma, osteogenic sarcoma, epithelial ovarian carcinoma, papillary adenocarcinoma, parathyroid tumor, pheochromocytoma, neuroblastoma, choriocarcinoma, myeloma, chordoma, neuroblastoma, carcinoma, pinealoma, plasmacytoma, retinoblastoma, rhabdomyosarcoma, sebaceous gland carcinoma, seminoma, skin carcinoma, melanoma, small cell lung carcinoma, squamous cell carcinoma, sweat gland carcinoma, synovioma, thyroid carcinoma, uveal melanoma, and wilms tumor.

Also described are methods of treating a hyperproliferative disorder in a mammal comprising co-administering to said mammal a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, and an antineoplastic agent. In some embodiments, the antineoplastic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors, biological response modifiers, anti-hormones, angiogenesis inhibitors, anti-androgens, SHIP activators-AQX-MN 100, Humax-CD20(ofatumumab), CD20 antagonists, IL 2-diphtheria toxin fusions.

The disease treated using the compounds, compositions, and methods described herein can be a hematological disorder. In certain embodiments, the hematologic disorder is selected from the group consisting of sickle cell anemia, Myelodysplastic Disorders (MDS), and myeloproliferative disorders. In a further embodiment, the myeloproliferative disease is selected from polycythemia vera, myelofibrosis, and essential thrombocythemia.

The compounds, compositions, and methods described herein can be used as anti-inflammatory agents with the added benefit of significantly less harmful side effects. The compounds, compositions, and methods described herein are useful for treating arthritis, including but not limited to rheumatoid arthritis, spondyloarthropathies, ankylosing spondylitis, gout, gouty arthritis, osteoarthritis, systemic lupus erythematosus, juvenile arthritis, acute rheumatic arthritis, enteropathic arthritis, neuropathic arthritis, psoriatic arthritis, and suppurative arthritis. The compounds, compositions, and methods described herein are also useful for treating osteoporosis and other related bone conditions. These compounds, compositions and methods described herein are also useful for treating gastrointestinal conditions such as reflux esophagitis, diarrhea, inflammatory bowel disease, crohn's disease, gastritis, irritable bowel syndrome and ulcerative colitis. The compounds, compositions, and methods described herein can also be used to treat pulmonary inflammation such as that associated with viral infections and cystic fibrosis. In addition, the compounds, compositions, and methods described herein are also useful in organ transplant patients, either alone or in combination with conventional immunomodulators. Further, the compounds, compositions and methods described herein are useful for treating pruritus and vitiligo. In particular, the compounds, compositions and methods described herein are useful for treating the particular inflammatory disease rheumatoid arthritis.

Other inflammatory diseases that may be prevented or treated include, without limitation: asthma, allergy, respiratory distress syndrome, or acute or chronic pancreatitis. In addition, respiratory diseases that may be prevented or treated include, but are not limited to, chronic obstructive pulmonary disease and pulmonary fibrosis. In addition, the MEK kinase inhibitors described herein are also associated with the production of prostaglandin endoperoxidase synthase-2 (COX-2). Proinflammatory mediators of the cyclooxygenase pathway derived from arachidonic acid, such as prostaglandins, are produced by inducible COX-2 enzymes. Modulation of COX-2 regulates these proinflammatory mediators, which affect a variety of cells and are important and key inflammatory mediators of a variety of disease states and conditions. In particular, these inflammatory mediators have been implicated in pain (e.g., sensitization of pain receptors) and edema. Thus, other MEK kinase-mediated conditions that may be prevented or treated include edema, analgesia, fever and pain (e.g. neuromuscular pain, headache, dental pain, arthritic pain and pain due to cancer).

Further, the disease treated by the compounds, compositions, and methods described herein can be an ophthalmic disorder. Ophthalmic diseases and other diseases where angiogenesis plays a role in pathogenesis, can be treated or prevented and include, but are not limited to: dry eye (including sjogren's syndrome), macular degeneration, angle-closure glaucoma and angle-opening glaucoma, retinal ganglionic degeneration, ocular ischemia, retinitis, retinopathy, uveitis, ocular photophobia, and inflammation and pain associated with acute injury to ocular tissue. The compounds, compositions, and methods described herein are also useful for treating post-operative inflammation or pain, such as that caused by ophthalmic surgery, e.g., cataract surgery and refractive surgery. In a further embodiment, the ophthalmic disorder is selected from dry eye, angle-closure glaucoma and angle-opening glaucoma.

In addition, the diseases treated by the compounds, compositions and methods described herein can be autoimmune diseases. Autoimmune diseases that may be prevented or treated include, but are not limited to: rheumatoid arthritis, inflammatory bowel disease, inflammatory pain, ulcerative colitis, crohn's disease, periodontal disease, temporomandibular joint disease, multiple sclerosis, diabetes, glomerulonephritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, graves' disease, hemolytic anemia, autoimmune gastritis, autoimmune neutropenia, thrombocytopenia, chronic active hepatitis, myasthenia gravis, atopic dermatitis, graft-versus-host disease and psoriasis. Inflammatory diseases that may be prevented or treated include, but are not limited to: asthma, allergy, respiratory distress syndrome or acute pancreatitis or chronic pancreatitis. In particular, the compounds, compositions and methods described herein are useful for treating the specific autoimmune diseases rheumatoid arthritis and multiple sclerosis.

Further, the disease treated by the compounds, compositions, and methods described herein can be a skin disorder. In certain embodiments, the skin condition is selected from the group consisting of, without limitation, melanoma, basal cell carcinoma, squamous cell carcinoma, and other non-epithelial skin cancers, as well as psoriasis and persistent pruritus, as well as other diseases associated with the skin and skin structure, which may be treated or prevented with MEK kinase inhibitors of the present invention.

Metabolic disorders that may be treated or prevented include, but are not limited to, metabolic syndrome, insulin resistance, and type I and type II diabetes. In addition, the compositions described herein may be used to treat insulin resistance and other metabolic disorders such as atherosclerosis that are typically associated with excessive inflammatory signals.

The compounds, compositions, and methods described herein are also useful in treating tissue damage in diseases such as vascular disease, migraine, adventitious arteritis nodosa, thyroiditis, aplastic anemia, hodgkin's disease, scleroderma (sclerodoma), rheumatic fever, type I diabetes, neuromuscular junction diseases (including myasthenia gravis), white matter diseases (including multiple sclerosis), sarcoidosis, nephritis, nephrotic syndrome, behcet's syndrome, polymyositis, gingivitis, periodontitis (periodontis), hypersensitivity reactions, post-injury swelling, ischemia (including myocardial ischemia, cardiovascular ischemia, and ischemia secondary to cardiac arrest), and the like. The compounds, compositions, and methods described herein may also be used to treat allergic rhinitis, respiratory distress syndrome, endotoxin shock syndrome, and atherosclerosis.

Further, the diseases treated by the compounds, compositions, and methods described herein can be cardiovascular conditions. In certain embodiments, the cardiovascular condition is selected from atherosclerosis, cardiac hypertrophy, idiopathic cardiomyopathy, heart failure, angiogenesis-related conditions or disorders, and proliferation following medical conditions including, but not limited to, restenosis due to surgery and angioplasty.

In addition, the diseases treated by the compounds, compositions, and methods described herein can be neurological disorders. In certain embodiments, the neurological disorder is selected from parkinson's disease, alzheimer's dementia, and central nervous system injury due to stroke, ischemia, and trauma. In other embodiments, the neurological disorder is selected from epilepsy, neuropathic pain, depression, and bipolar disorder.

In addition, the disease treated by the compounds, compositions, and methods described herein can be cancer, such as acute myeloid leukemia, thymus, brain, lung, squamous cell, skin, eye, retinoblastoma, intraocular melanoma, oral and oropharyngeal cancer, bladder, stomach (gastic), stomach (stomaching), pancreas, bladder, breast, cervix, head, neck, kidney, liver, ovary, prostate, colorectal, esophagus, testis, gynecological, thyroid, CNS, PNS, AIDS-related (e.g., lymphoma and kaposi's sarcoma), or virus-induced cancer. In some embodiments, the compounds and compositions are used to treat non-cancerous hyperproliferative disorders, such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., Benign Prostatic Hypertrophy (BPH)).

In addition, the diseases treated by the compounds, compositions and methods described herein can be pancreatitis, kidney diseases (including proliferative glomerulonephritis and nephropathy due to diabetes), pain, diseases associated with angiogenesis or vasculogenesis, tumor angiogenesis, chronic inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease, atherosclerosis, skin diseases (such as psoriasis, eczema, and scleroderma), diabetes, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, hemangioma, tendonitis, bursitis, sciatica, glioma, melanoma, kaposi's sarcoma, and ovarian, breast, lung, pancreatic, prostate, colon, and epidermoid cancers in mammals.

In addition, the disease treated by the compounds, compositions, and methods described herein can be prevention of blast implantation in a mammal.

Patients that may be treated with a compound described herein, or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate, or derivative thereof, according to the methods of the invention include, for example, patients who have been diagnosed with a disease comprising: psoriasis; restenosis; atherosclerosis; BPH; breast cancer, such as ductal carcinoma in ductal tissue in the breast, medullary carcinoma, jelly-like carcinoma, tubular carcinoma, and inflammatory breast cancer; ovarian cancer (including epithelial ovarian tumors such as adenocarcinoma in the ovary and adenocarcinoma that has metastasized from the ovary into the abdominal cavity); uterine cancer; cervical cancer, such as adenocarcinoma (including squamous cell carcinoma and adenocarcinoma) in the cervical epithelium; prostate cancer, such as prostate cancer selected from: adenocarcinoma that has metastasized to bone; pancreatic cancer, such as epithelioid cancer in pancreatic ductal tissue and adenocarcinoma in pancreatic duct; bladder cancer, such as transitional cell carcinoma in the urinary bladder, urothelial cancer (transitional cell carcinoma), tumors in urothelial cells in the bladder lining, squamous cell carcinoma, adenocarcinoma, and small cell cancer; leukemias such as Acute Myeloid Leukemia (AML), acute lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, and myeloproliferative disorders; bone cancer; lung cancer such as non-small cell lung cancer (NSCLC), which is classified into squamous cell carcinoma, adenocarcinoma, and large-cell undifferentiated carcinoma, and small-cell lung cancer; skin cancers such as basal cell carcinoma, melanoma, squamous cell carcinoma, and actinic keratosis (which is a skin disease that sometimes progresses to squamous cell carcinoma); retinoblastoma of the eye; cutaneous or intraocular (ocular) melanoma; primary liver cancer (cancer originating in the liver); kidney cancer; thyroid cancer such as papillary thyroid cancer, follicular thyroid cancer, medullary thyroid cancer, and thyroid cancer polymorphous; AIDS-related lymphomas such as diffuse large B-cell lymphoma, B-cell immunoblastic lymphoma, and small anucleated cell lymphoma; kaposi's sarcoma; viral-induced cancers (including Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), and hepatocellular carcinoma); human lymphotropic virus type 1 (HTLV-1) and adult T-cell leukemia/lymphoma; and Human Papilloma Virus (HPV) and cervical cancer; central nervous system Cancers (CNS) such as primary brain tumors, including gliomas (astrocytomas, glioblastoma multiforme, or glioblastoma multiforme), oligodendrogliomas, ependymomas, meningiomas, lymphomas, schwannoma, and medulloblastoma; peripheral Nervous System (PNS) cancers such as auditory neuroma and Malignant Peripheral Nerve Sheath Tumor (MPNST) including neurofibroma and nerve sheath tumor, malignant fibrous cell tumor, malignant fibrous histiocytoma, malignant meningioma, malignant mesothelioma, and malignant mixed muller tumor; oral and oropharyngeal cancers such as hypopharynx, larynx, nasopharynx, and oropharynx; stomach cancer such as lymphoma, gastric stromal tumor, and carcinoid tumor; testicular cancers such as Germ Cell Tumors (GCTs), which include seminoma and non-seminoma and gonadal stromal tumors, which include leydig cell tumors and serituximab; thymus cancer such as thymoma, thymus cancer, hodgkin's disease, non-hodgkin's lymphoma-like cancer or carcinoid tumor; rectal cancer; and colon cancer.

Medicine box

The compounds, compositions, and methods described herein provide kits for treating conditions such as those described herein. These kits comprise in a container one or more compounds or compositions described herein, and optionally instructions for using the kit according to the various methods and procedures described herein. Such kits may also include information such as scientific references, drug instruction materials, clinical trial results, and/or summaries of these, etc., which describe or demonstrate the activity and/or advantages of the compositions, and/or which describe dosages, methods of administration, side effects, drug interactions, or other information useful to the healthcare provider. Such information can be based on the results of various studies, for example, studies using experimental animals, including in vivo models as well as studies based on human clinical trials. The kits described herein can be provided, sold and/or marketed to medical providers, including physicians, nurses, pharmacists, prescription officials, and the like. In some embodiments, the kit may also be sold directly to the consumer.

The compounds described herein may be used in diagnostics and as research reagents. For example, the compounds described herein, alone or in combination with other compounds, can be used as tools in differential and/or combinatorial assays to elucidate the expression pattern of genes expressed in cells and tissues. As a non-limiting example, the expression pattern in cells or tissues treated with one or more compounds is compared to the expression pattern in control cells or tissues not treated with the compound, and the resulting patterns are analyzed to determine differential levels of gene expression as they relate to, for example, disease association, signaling pathways, cellular localization, detecting expression levels, size, structure or function of genes. These assays can be performed on stimulated or unstimulated cells, and in the presence or absence of other compounds that affect the expression pattern.

In addition to use in human therapy, the compounds and formulations of the present invention are also useful in veterinary therapy of companion animals (e.g., dogs, cats), exotic animals, and livestock (e.g., horses), including mammals, rodents, and the like.

The examples and methods of preparation provided below further illustrate and exemplify the compounds of the present invention and methods of making such compounds. It should be understood that the scope of the present invention is by no means limited to the scope of the following examples and preparation methods.

Examples

General exemplary procedure for the Synthesis of sulfonamides

Step A: to a solution of amine (1eq) in anhydrous dichloromethane (3mL/mmole) was added anhydrous triethylamine (5 eq). To this solution was added sulfuryl chloride (1eq) and the solution was stirred at room temperature for 16 h. The solvent was evaporated and the residue was purified by flash column chromatography over silica gel.

And B: to a stirred solution (5ml/mmole) of amine (1eq) in anhydrous pyridine was added sulfonyl chloride (1-5 eq). The reaction mixture was stirred at 40 ℃ for 48 hours. The reaction mixture was partitioned with water and EtOAc. Washed with brine and dried (MGSO)₄) And concentrated under reduced pressure. The residue was purified by flash column chromatography over silica gel.

And C: substitution of iodine atom:

in a microwave reactor, 1eq. aryl iodide, 1.5eq. boric acid or borate, 0.25eq. pdcl contained in a deoxygenated mixture of dioxane and water (3:1), are reacted ₂(dppf) xDCM and 10eq. Anhydrous K₂CO₃The suspension of the powder was heated at 115 ℃ for 60 min. By aq₄Cl/THF extraction and Na₂SO₄The organic portion is dried. Flash column chromatography (Si, EtOAc/hexanes, or CHCl) was used₃MeOH) purified the crude reaction product. Yield: 20 to 40 percent.

Step D: synthesis of N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2- (alkylamino) ethanesulfonamide:

2-chloro-ethanesulfonyl chloride (0.1ml,1mmol) was added to 5, 6-difluoro-N¹- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.364g,1mmol) and triethylamine (0.28ml,2mmol) in CH₂Cl₂(5ml) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture is then treated with an excess of amine (10eq) in solution or neat liquid. The reaction mixture was stirred at room temperatureAnd 6 h. By CH₂Cl₂The reaction mixture was diluted (10ml) and water (10 ml). The organic layer was successively diluted HCl (2X20ml,2N) and saturated NaHCO₃(2 × 10ml) solution wash. Then dried (MgSO)₄)CH₂Cl₂Layer and evaporated to give crude product. Purification of the impure product under preparative HPLC conditions gave the pure product in 50-60% yield.

Example 1

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) methanesulfonamide:

step A:2, 3-difluoro-N- (2-fluoro-4-iodophenyl) -6-nitroaniline ：

To a solution of 2-fluoro-4-iodoaniline (11.40g,47mmol) in 100ml dry THF was added dropwise a 1M solution of LHMDS in THF (47mmol) (47ml) at 0 ℃. The color of the solution turned dark purple. The solution was transferred via cannula to a dropping funnel and this solution (containing the amine free base) was added portionwise to a solution of 2,3, 4-trifluoronitrobenzene (8.321g,47.0mmol) in anhydrous THF (50ml) at 0 ℃. After the addition was complete, the mixture was stirred at room temperature under argon for 15 hours. The volume of the solvent was reduced and then extracted with ethyl acetate and brine. The organic layer was dried over sodium sulfate, the solvent removed, and purified by flash chromatography (EtOAc/hexane 1:5, R)_f=0.58) the resulting dark oil was purified to give crude product which was dried in vacuo to brown solid (yield: 6.23g, 33.6%): m/z =393[ M-1 ]]^-。

And B:5，6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1，2-diamines：

To a solution of nitro-diarylamine (6.23g,15.8mmol) in 300ml ethanol were added iron powder (13.74g,246mmol) and ammonium chloride (13.59g,254mmol) and the mixture was heated at 100 ℃ oil bath temperature with stirring for 14 hours. Filter and wash the residue twice with ethanol. The ethanol was removed in vacuo and the residue was extracted with ethyl acetate/1M NaOH solution. During the extraction, more precipitate formed was filtered and discarded. The combined organic layers were washed with brine and dried over sodium sulfate. Removing the solvent and separating from CHCl ₃The crude product was recrystallized from hexane (1: 50). The brown needle-like product (2.094g,66%,), R_f=0.44(EtOAc/Hex1:3)。¹H-NMR(500MHz,CDCl₃),=7.40-7.38(dd,1H,J=11.3Hz,J=1.5Hz),7.25-7.23(d,1H,J=8.5Hz),6.97-6.92(q,1H,J=9Hz),6.51-6.48(m,1H),6.24-6.21(t,1H,J=9Hz),5.3(s,1H,NH,br),3.80(s,2H,NH₂,br),LRMS(ESI):m/z=365[M+H]^-。

And C:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) methanesulfonamide：

Following general procedure a, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with methanesulfonyl chloride to afford the desired product.¹HNMR：(500MHz,CDCl₃)：=7.38-7.37(d,1H),7.35-7.34(m,1H),7.27-7.26(m,1H),7.20-7.0(q,1H),6.68(s,1H,br),6.15-6.12(q,1H),5.65(s,1H,br),2.95(s,3H)；m/z=441[M-1]^-。

Example 2

2-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropanesulfonamide:

following general procedure a, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with cyclopropanesulfonyl chloride to give the desired product.¹HNMR：(500MHz,CDCl₃)：=7.38-7.37(d,1H),7.35-7.34(m,1H),121-1Id(m,1H),7.20-7.0(q,1H),6.68(s,1H,br),6.15-6.12(q,1H),5.65(s,1H,br),3.25-3.20(m,1H),2.4-2.3(m,2H),2.0-1.8(m,2H)；m/z=467[M-1]^-。

Example 3

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) propane-2-sulfonamide:

the desired product was obtained according to general procedure a, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with isopropylsulfonyl chloride. Yield: 39 percent.¹H-NMR(500MHz,CDCl₃)：=7.50-7.43(m,1H),7.35-7.34(m,1H),7.27-7.26(m,1H),7.15-7.09(q,1H,J=1.6Hz),6.62(s,1H,br),6.22-6.18(q,1H,J=1.5Hz),5.65(s,1H,br),3.30-3.28(m,1H),1.38-1.37(d,6H,J=1.2Hz)；m/z=469[M-I]^-。

Example 4

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) butane-l-sulfonamide:

the desired product was obtained according to general procedure a, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with N-butylsulfonyl chloride. Yield: and 55 percent.¹H-NMR(500MHz,CDCl₃)：=7.50-7.43(m,1H),7.35-7.34(m,1H),7.27-7.26(m,1H),7.15-7.09(q,1H,J=1.6Hz),6.62(s,1H,br),6.22-6.18(q,1H,J=1.5Hz)55.65(s,1H,br),3.06-3.031(t,2H,J=1.4Hz),1.75-1.71(m,2H),1.38-1.36(m,2H),0.87-0.86(t,3H,J=1.3Hz)；m/z=483[M-1]^-。

Example 5

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2,2, 2-trifluoroethylsulfonamide:

the desired product was obtained according to general procedure a, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 1,1, 1-trifluoroethylsulfonyl chloride. Yield: 28 percent. M/z =509[ M-1 ]^-。

Example 6

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) butane-2-sulfonanilide:

following general procedure a, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with sec-butylsulfonyl chloride to give the desired product. Yield: 22 percent.¹H-NMR(500MHz,MeOH[d4])：=7.60-7.40(m,3H),7.18-7.00(q,1H),6.55-6.45(m,1H),3.55-3.50(m,1H),2.20-2.00(m,1H),1.80-1.60(m,1H),1.43-1.40(d,3H),1.06-1.04(t,3H)；m/z=483[M-1]^-。

Example 7

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -N-methylcyclopropanesulfonamide:

to a solution of N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-sulfonamide (see example 2) (283.9mg,0.61mmol) in 3ml of anhydrous THF was added a 1M solution of LHMDS (0.6ml,0.6mol) at-78 ℃ and the solution was stirred at this temperature for 10 min. Methyl iodide (0.8ml,1.824g,12.9mmol) was then added and the mixture allowed to warm to room temperature and stir for 7 h. The solvent was removed and the residue was extracted with EtOAc and brine. With Na₂SO₄The organic portion was dried and the solvent was removed. Flash column chromatography (Si, EtOAc/hexanes 1:2, R)_f=0.45) purifying the crude product obtained. Yield: (205mg, 70%).¹H-NMR(500MHz,CDCl₃)：=7.41-7.39(d,1H,J=10Hz),7.30-7.29(d,1H,J=8.0Hz),7.23-7.20(m,1H),6.98-6.93(q,1H,J=8.5Hz),6.60(s,1H,br),6.51-6.47(m,1H),3.23(s,3H),2.46-2.42(m,1H),1.19-1.16(m,2H),1.04-1.02(m,2H)；m/z=481[M-1]^-。

Example 8

l-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) methanesulfonamide:

following general procedure a, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with chloromethylsulfonyl chloride gave the desired product, M/z =475[ M-1 [ ]^-。

Example 9

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2-methylpropane-2-sulfonamide:

the desired product was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 2-methylpropane-2-sulfonyl chloride (synthesized according to literature procedures).¹H NMR(300MHz,CDCl₃)：7.50(m,1H),7.43(dd,J=1.8&10.5Hz,1H),7.28(br s,1H),7.10(dd,J=9.0&17.7Hz,1H),6.48(br s,D₂O exchangeable, 1H),6.19(t, J = 7.8)&9.6Hz,1H),5.58(br s,D₂O may be exchanged, 1H),1.39(s, 9H); m/z =383[ M-1 ]]^-。

Example 10

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopentane sulfonamide:

following general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with cyclopentanesulfonyl chloride to give the desired product.¹H NMR(300MHz,CDCl₃)：7.42(dd,J=2.1&10.5Hz,1H),7.36(ddd,J=2.4,4.8,&9.3Hz,1H),7.25(m,2H),7.10(dd,J=9.6&17.7Hz,1H),6.67(br s,D₂O exchangeable, 1H),6.20(dt, J =1.5, 8.4)&17.4Hz,1H),3.53(p,1H),1.80(m,8H)；m/z=495[M-1]^-。

Example 11

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclohexanesulfonamide:

according to the general procedure B, 5,6difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with cyclohexanesulfonyl chloride to give the desired product.¹H NMR(300MHz,CDCl₃)：7.43(dd,J=1.5&10.2Hz,1H),7.37(ddd,J=2.4,4.8&9.6Hz,1H),7.27(m,1H),7.11(dd,J=9.3&18.0Hz,1H),6.64(br s,1H),6.18(dt,J=1.5,9.0&17.4Hz,1H),5.63(br s,1H),2.95 (three sets of triplets, 2.10-1.16(m, 10H); m/z =509[ M-1]^-。

Example 12

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l-methylcyclopropane-l-sulfonamide:

step A:n-butyl 3-chloro-1-propanesulfonate:

will be in CH₂Cl₂Triethylamine (28ml,200mmol) in (50ml) was slowly added to 3-chloro-1-propanesulfonyl chloride (36.6g,200mmol) and 1-butanol (18.4g,240 mmol) in CH ₂Cl₂(250ml) in ice-cold solution and stirring was continued for 16 h. By CH₂Cl₂The mixture was diluted (200ml), washed (aqueous HCl) and dried (MgSO)₄) And the solvent was evaporated to give the title product 1(40.85g,95%) as a yellowish oil in crude form, which was used in the next reaction without further purification.¹H NMR(CDCl₃₎)0.94(t, J =7.5Hz,3H),1.44 (hexameric peak, 2H),1.72 (quintuple peak, 2H),2.31 (quintuple peak, 2H),3.27(t, J =6.9Hz,2H),3.68(t, J =6.3Hz),4.23(t, J =6.6Hz, 2H).

And B:1-butyl cyclopropanesulfonate:

3-chloro-l-substituted benzene at-78 ℃ under nitrogen atmosphere1-butyl propanesulfonate solution (4.6g,21.39mmol in 25ml THF) and butyllithium solution (14.7ml,23.53mmol,1.6M, THF) were added simultaneously to THF (150 ml). The solution was warmed to 0 ℃ and then quenched with (2 ml). Evaporate the volatiles under reduced pressure and use CH₂Cl₂The residue was extracted (150 ml). The extract was washed with water and dried (MgSO)₄) And evaporated to give the crude desired product (3.23g,78.22%) as a pale yellow oil in almost pure form, which was used in the next step without further purification.¹H NMR(300MHz,CDCl₃)0.94(t, J =7.5Hz,3H),1.07(m,2H),1.25(m,2H),1.45 (hexameric peak, 2H),1.74 (pentaric peak, 2H),2.45 (heptaric peak, 1H),4.23(t, J =6.6Hz, 2H).

And C:l-methyl-cyclopropanesulfonic acid butyl ester:

to a solution of 1-butyl cyclopropanesulfonate (1g,5.58mmol) in THF (15ml) was slowly added a solution of butyllithium (3.84ml,6,14mmol,1.6M, THF) at-78 deg.C under a nitrogen atmosphere. After 15 min, MeI (0.72ml,11.16mmol) was added, the solution was warmed to 0 ℃ and quenched with water (1 ml). Evaporate the volatiles under reduced pressure and use CH₂Cl₂The residue was extracted (100 ml). The extract was washed with water and dried (MgSO)₄) And evaporated. The residue is chromatographed on silica gel (eluent: hexane/CH)₂Cl₂) Purification gave the title product as a colorless oil (0.59g, 55.0%).¹H NMR(300MHz,CDCl₃))0.84(m,2H),0.95(t,J=7.2Hz,3H),1.43(m,4H),1.53(s,3H),1.74(m,2H),4.21((t,J=6.6Hz,2H)。

Step D:potassium 1-methyl-cyclopropanesulfonate:

1-butyl 1-methyl-cyclopropanesulfonateA mixture of (0.386g,2mmol) and potassium thiocyanate (0.194g,2mmol) in DME (5ml) and water (5ml) was refluxed for 16 h. The volatiles were evaporated to give the crude sulfonate salt (0.348g, quantitative), which was dried under vacuum at 50 ℃ for 16 h. This crude product was used in the next reaction without further purification.¹H NMR(300MHz,D₂O)0.56(t,J=6.3Hz,2H),0.96(t,J=6.3Hz,2H),1.26(s,3H)。

Step E:l-methyl-cyclopropanesulfonyl chloride:

a solution of potassium 1-methyl-cyclopropanesulphonate (0.348g,2mmol), thionyl chloride (5ml) and DMF (5 drops) was refluxed at 60 ℃ for 16 h. Evaporate the volatiles under reduced pressure and use CH ₂Cl₂The residue was extracted (50 ml). The extract was washed with water and dried (MgSO)₄) And evaporated to give a crude product as a yellow gummy oil which was used in the next reaction without further purification.

Step F:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1-methylcyclopropane-1-sulfonamide:

the desired product was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 1-methyl-cyclopropanesulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.42(dd,J=1.8&10.5Hz,1H),7.36(ddd,J=2.4,4.5&9.0Hz,1H),7.27(d,J=6.0Hz,1H),7.07(dd,J=9.3&17.7Hz,1H),6.24(dt,J=2.1,8.7&17.4Hz,1H),5.86(br s,1H),1.43(s,3H),1.33(t,J=5.4Hz,2H),0.75(dd,J=5.1&6.3Hz,2H)；m/z=481[M-1]^-。

Example 13

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-l-sulfonamide:

step A:butyl cyclopropanesulfonate:

cyclopropanesulfonyl chloride (5g,35mmol,1eq) was dissolved in excess BuOH (20mL), the reaction mixture was cooled at-10 ℃ and pyridine (5.8mL,70mmol,2eq) was slowly added dropwise. The mixture was allowed to warm slowly to room temperature and stirred overnight. The solvent was removed under reduced pressure and the resulting white solid was dissolved in CHCl₃In (1). The organic phase was washed with water, brine, dried (MgSO4), and concentrated to give an oil (4.8g,24.9mmol, 71%).¹H NMR(300MHz,CDCl₃)：4.25(t,2H),2.46(m,1H),1.74(m,2H),1.45(m,2H),1.25(dd,2H),1.09(dd,2H),.93(t,3H)。

And B:1-allylcyclopropane-1-sulfonic acid butyl ester:

to a solution of 1-butyl cyclopropanesulfonate (4.8g,24.9mmol) in THF was added a solution of butyllithium (15.6ml,24.9mmol,1.6M, THF) and allyl iodide (24.9mmol) simultaneously at-78 deg.C under a nitrogen atmosphere. The reaction mixture was stirred at-78 ℃ for 2 hours and at room temperature for 3 hours. Evaporate the volatiles under reduced pressure and use CH ₂Cl₂The residue was extracted (100 ml). The extract was washed with water and dried (MgSO)₄) And then evaporated. The residue is chromatographed on silica gel (eluent: hexane/CH)₂Cl₂) Purification gave the title product as a colorless oil (3.75g, 69.0%).¹H NMR(300MHz,CDCl₃)：5.6(m,1H),5.13-5.08(t,2H),4.21(t,2H),2.65(d,2H),1.7(m,2H),1.4(m,4H),.93(m,5H)。

And C:l-allylcyclopropane-l-sulfonic acid potassium:

a mixture of 1-butyl l-methyl-cyclopropanesulphonate (3.75g,17.2mmol) and potassium thiocyanate (1.7g,17.2mmol) in DME (20ml) and water (20ml) was refluxed for 16 h. The volatiles were evaporated to give the crude sulfonate (3.44g, quantitative), which was dried under vacuum at 50 ℃ for 16 h. This crude product was used in the next reaction without further purification.¹H NMR(CDCl₃)：5.6(m,1H),4.91-4.85(dd,2H),2.471-2.397(d,2H),0.756(m,2H),0.322(m,2H)。

Step D:l-allylcyclopropane-l-sulfonyl chloride:

a solution of potassium l-allylcyclopropane-l-sulfonate (3.44g,17.2mmol), thionyl chloride (10ml) and DMF (5 drops) was refluxed at 60 ℃ for 16 h. Evaporate the volatiles under reduced pressure and use CH₂Cl₂The residue was extracted (50 ml). The extract was washed with water and dried (MgSO)₄) And evaporated to give the crude product as a yellow gummy oil which was washed with hexane and used in the next reaction without further purification (2.7g,15mmol, 87%).¹H NMR(300MHz,CDCl₃)：5.728(m,1H),5.191(t,2H),2.9(d,2H),0.756(m,2H),0.322(m,2H)。

Step E:l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-1-sulfonamide:

According to the general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl)Reaction of benzene-1, 2-diamine with 1-allylcyclopropane-l-sulfonyl chloride gives the desired product. M/z =507[ M-1 ]]^-。

Step F:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1- (2, 3-dihydroxypropyl) cyclopropyl Alkane-1-sulfonamides:

1-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-1-sulfonamide (0,77g,1.52mmol) and 4-methylmorpholine N-oxide (0,18g,1.52mmol) were dissolved in THF (50 mL). Osmium tetroxide (0.152mmol,0.965mL,4% in H) was added at room temperature₂O) and the reaction mixture was stirred at room temperature for 16 hours. EtOAc was added and the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product (0.65g, 79%).¹H NMR(300MHz,CDCl₃+D₂O)：7.38(dd,J=1.8&10.5Hz,1H),7.36(ddd,J=2.4,5.1&9.3Hz,1H),7.25(d,J=8.7Hz,1H),7.02(dd,J=9.0&17.7Hz,1H),6.27(dt,J=3.0,8.7&17.4Hz,1H),3.92(m,1H),3.54(dd,J=3.9&11.1Hz,1H),3.39(dd,J=6.6&11.1Hz,1H),2.16(dd,J=9.6&15.9Hz,1H),1.59(d,J=14.1Hz,1H),1.41(m,1H),1.26(m,1H),0.83(m,2H)；m/z=542[M-1]^-。

Example 14

(S) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-l-sulfonamide:

the racemic mixture (example 13) was separated by chiral HPLC to give the pure S isomer.¹H NMR(300MHz,CDCl₃+D₂O)：7.38(dd,J=1.8&10.5Hz,1H),7.36(ddd,J=2.4,5.1&9.3Hz,1H),7.25(d,J=8.7Hz,1H),7.02(dd,J=9.0&17.7Hz,1H),6.27(dt,J=3.0,8.7&17.4Hz,1H),3.92(m,1H),3.54(dd,J=3.9&11.1Hz,1H),3.39(dd,J=6.6&11.1Hz,1H),2.16(dd,J=9.6&15.9Hz,1H),1.59(d,J=14.1Hz,1H),1.41(m,1H),1.26(m,1H),0.83(m,2H)；m/z=542[M-1]^-。

Example 15

(R) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-l-sulfonamide:

the racemic mixture (example 13) was separated by chiral HPLC to give the pure R isomer. ¹H NMR(300MHz,CDCl₃+D₂O)：7.38(dd,J=1.8&10.5Hz,1H),7.36(ddd,J=2.4,5.1&9.3Hz,1H),7.25(d,J=8.7Hz,1H),7.02(dd,J=9.0&17.7Hz,1H),6.27(dt,J=3.0,8.7&17.4Hz,1H),3.92(m,1H),3.54(dd,J=3.9&11.1Hz,1H),3.39(dd,J=6.6&11.1Hz,1H),2.16(dd,J=9.6&15.9Hz,1H),1.59(d,J=14.1Hz,1H),1.41(m,1H),1.26(m,1H),0.83(m,2H)；m/z=542[M-1]^-。

Example 16

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l- (2-hydroxyethyl) cyclopropane-l-sulfonamide:

step A:2- (1-bromocyclopropyl) ethanol:

to a solution of pure diethylzinc (3.3ml,3.977g,30mmol) in 100ml of anhydrous DCM was added dropwise very slowly to trifluoroacetic acid (2.31ml,3.4188g,30 mmol). (Note: vigorous gas evolution, exotherm!). After TFA addition was complete, the suspension was stirred at the same temperature for 20min, then diiodomethane (2.45ml,8.134g,30.4mmol) was added. Stirring is continued for a further 20min at 0 ℃ and then a solution of 3-bromobut-3-en-l-ol (1ml,1.523g,10.1mmol) in 10ml DCM is added at the same temperature. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction of this mixture was quenched with 100ml MeOH and 40ml brine, which was then stirred for an additional 30 min. Reducing solvent, and using CHCl₃/aq.NH₄The residue was extracted with Cl. The organic layer was collected, washed with brine and water, and the solvent was removed to yield 2- (l-bromocyclopropyl) -ethanol (1.6564g,100%) of sufficient purity.¹H-NMR(500MHz,CDCl₃)：=3.90-3.83(t,2H),1.91-1.87(t,2H),1.71(s,1H,br),1.14-1.09(m,2H),0.83-0.79(m,2H)。

And B:TBS protected 2- (l-bromocyclopropyl) ethanol:

to a solution of cyclopropyl alcohol (step A) (1.303g,7.95mmol) in 30ml of anhydrous DCM were added anhydrous pyridine (1.2ml,1,1736g,14.8mmol) and TBSOTf (2.7ml,3.1077g,11.76mol), and the solution was stirred at room temperature for 16 h. With CHCl ₃Extracted with brine and MgSO₄The organic portion is dried. Reducing solvent and performing flash column chromatography (Si, CHCl)₃Hexane 1:10, R_f=0.4) purifying the crude product. Yield: 0.796g, 36%.¹H-NMR(500MHz,CDCl₃)：=3.95-3.75(t,2H),1.95-1.85(t,2H),1.15-1.05(m,2H),0.95-0.80(m,HH),0.15-0.05(s,6H)。

And C:TBS protected 2- (1-chlorosulfonyl cyclopropyl) ethanol:

to a solution of cyclopropyl bromide prepared in step B (1.1227g,4.04mmol) in 15ml of anhydrous diethyl ether at-78 deg.C was added a 1.7M solution of t-BuLi in pentane (4.8ml,8.16 mmol). The solution was stirred at this temperature for 30min and then transferred via a transfer cannula (transfer canola) at-78 ℃ to a solution of freshly distilled sulfonyl chloride (0.65ml,1.029g,8.1mmol) in 8ml diethyl ether. The yellow suspension was allowed to warm to room temperature. The solvent was removed and the residue was dried in vacuo to remove excess sulfonyl chloride. The residue was then extracted twice with hexane, filtered and the solvent evaporated in vacuo to yield sulfonyl chloride as a colorless oil of sufficient purity. Yield: 870mg (72%).¹H-NMR(300MHz,CDCl₃)：=3.95-3.85(t,2H),2.35-2.25(t,2H),1.80-1.70(m,2H),1.45-1.38(m,2H),0.90(s,9H),0.10(s,6H)。

Step D:TBS protected N- (3.4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1- (2-hydroxyethyl) Cyclopropane-1-sulfonamides:

the desired product was obtained according to general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with cyclopropylsulfonyl chloride prepared in step C. ¹H-NMR(300MHz,CDCl₃)：=7.44-7.39(dd,1H),7.32-7.24(m,2H),7.1-6.98(q,1H),6.34-6.24(m,1H),6.16(s,1H,br),3.85-3.75(t,2H),2.15-2.00(t,2H),1.35-1.20(m,2H),0.95-0.75(m,11H),0.10(s,6H)；m/z=625[M-1]^-。

Step E:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l- (2-hydroxyethyl) cyclopropane-l- Sulfonamide:

at 0 deg.CTo a solution of the TBS protected sulfonamide prepared in D (21mg,0.033mmol) in 1ml THF was added 0.1ml aq.1.2N HCl solution and the solution was stirred for 2 h. Reducing solvent and using NaHCO₃Aqueous and EtOAc extract the residue. With MgSO₄The organic portion was dried and the volatiles were removed. By flash column chromatography (Si, CHCl)₃/MeOH10:1,R_f=0.45) purification of the crude product yielded a pure product. Yield: 16.9mg (100%).¹H-NMR(300MHz,CDCl₃)：=7.44-7.39(dd,1H),7.32-7.24(m,2H),7.1-6.98(q,1H),6.34-6.24(m,1H),6.16(s,1H,br),3.85-3.75(t,2H),2.15-2.00(t,2H),1.35-1.20(m,2H),0.95-0.85(m,2H)；m/z=511[M-1]^-。

Example 17

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -3-hydroxypropane-l-sulfonamide:

to 3-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -propane-1-sulfonamide (69.4mg,0.138mmol) in 8ml1, 4-dioxane and 2ml H₂To a solution in a mixture of O was added KOH powder (0.674g,12.0mmol) and the mixture was heated to reflux temperature for 3 days. Extracted with EtOAc/brine, over Na₂SO₄The organic portion was dried and the volatiles were removed. Flash column chromatography (Si, DCM/MeOH5:1, R)_f=0.3) purification of the residue. Yield: 41mg (62%).¹H-NMR(500MHz,MeOH[d4])：=7.38-7.21(d,1H),7.23-7.21(d,1H),7.06-7.00(q,1H),6.52-6.50(m,1H),6.17-6.13(t,1H),3.30-3.27(t,2H),2.86-2.83(t,2H),2.05-2.00(m,2H)；m/z=485[M-1]^-。

Example 18

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2-methyl-5- (trifluoromethyl) furan-3-sulfonamide:

According to general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 2-methyl-5- (trifluoromethyl) furan-3-sulfonyl chloride (0.5mmol) to form N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2-methyl-5- (trifluoromethyl) furan-3-sulfonamide.¹H NMR(CDCl₃)2.2(s,3H),5.3(s,1H),6.0(dt,1H),6.8(s,1H),6.95(s,1H),7.0-7.3(m,3H),7.4(dd,1H)。

Example 19

N- (5- (N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) sulfamoyl) -methylthiazol-2-yl) acetamide:

following general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 2-acetamido-4-methylthiazole-5-sulfonyl chloride (0.5mmol) to give N- (5- (N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) sulfamoyl) -4-methylthiazol-2-yl) acetamide.¹H NMR(CDCl₃))2.1(s,3H),2.2(s,3H),5.9(dt,1H),6.05(s,1H),7.0-7.6(m,3H),7.4(dd,1H),8.0(s,1H)。

Example 20

5- (5-chloro-1, 2, 4-thiadiazol-3-yl) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-2-sulfonamide:

according to general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 5- (5-chloro-1, 2, 4-thiadiazol-3-yl) thiophene-2-sulfonyl chloride (0.5mmol) to give 5- (5-chloro-1, 2, 4-thiadiazol-3-yl) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-2-sulfonamide.¹H NMR(300MHz,CDCl₃))5.8(dt,1H),5.95(s,1H),6.95(d,1H),7.4(m,2H),7.6(d,1H),7.8(s,1H)。

Example 21

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -3,5 dimethylisoxazole-4-sulfonamide:

According to general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 3, 5-dimethylisoxazole-4-sulfonyl chloride (0.5mmol) to give N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -3, 5-dimethylisoxazole-4-sulfonamide.¹H NMR(300MHz,CDCl₃))2.2(s,3H),2.4(s,3H),5.8(s,1H),6.0(dt,1H),5.95(s,1H),6.9(s,1H),7.0(q,1H),7.2(m,3H),7.4(dd,1H)。

Example 22

5-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1, 3-dimethyl-lH-pyrazole-4-sulfonamide:

according to the general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 5-chloro-1, 3-dimethyl-lH-pyrazole-4-sulfonyl chloride (0.5mmol) to give 5-chloro-N- (3, 4-difluoro-2-(2-fluoro-4-iodophenylamino) phenyl) -1, 3-dimethyl-lH-pyrazole-4-sulfonamide.¹H NMR(300MHz,CDCl₃))2.1(s,3H),3.6(s,3H),5.8(s,1H),5.95(dt,1H),7.0(q,1H),7.2(d,1H),7.3(m,2H),7.4(dd,1H)。

Example 23

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2, 5-dimethylfuran-3-sulfonamide:

following general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 2, 5-dimethylfuran-3-sulfonyl chloride (0.5mmol) to give N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2, 5-dimethylfuran-3-sulfonamide.¹H NMR(300MHz,CDCl₃))2.2(s,3H),2.3(s,3H),5.8(s,1H),6.0(dt,1H),6.8(s,1H),7.0(q,1H),7.2(d,1H),7.3(m,2H),7.4(dd,1H)。

Example 24

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l-methyl-3- (trifluoromethyl) -lH-pyrazole-4-sulfonamide:

according to general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with l-methyl-3- (trifluoromethyl) -lH-pyrazole-4-sulfonyl chloride (0.5mmol) to give N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -l-methyl-3- (trifluoromethyl) -lH-pyrazole-4-sulfonamide. ¹H NMR(300MHz,CDCl₃))3.8(s,3H),5.7(s,1H),6.0(dt,1H),7.0(q,1H),7.2(m,2H),7.4(dd,1H),7.8(s,1H)。

Example 25

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2, 4-dimethylthiazole-5-sulfonamide:

following general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine (0.182mmol) was reacted with 2, 4-dimethylthiazole-5-sulfonyl chloride (0.5mmol) to give N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2, 4-dimethylthiazole-5-sulfonamide.¹H NMR(300MHz,CDCl₃))2.3(s,3H),2.6(s,3H),5.7(s,1H),5.9(dt,1H),7.1(q,1H),7.2(d,1H),7.3(m,IH),7.4(d,1H),7.4(s,1H)。

Example 26

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1, 2-dimethyl-lH-imidazole-4-sulfonamide:

following general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with 1, 2-dimethyl-lH-imidazole-4-sulfonyl chloride to give the title compound.¹H NMR(300MHz,CDCl₃)：7.95(br s,1H),7.37(dd,J=1.8&10.8Hz,1H),7.32-7.14(m,3H),6.98(dd,J=9.6&17.7Hz,1H),5.87(dt,J=4.2,9.0&17.4Hz,1H),5.55(br s,1H),3.49(s,3H),2.31(s,3H)。

Example 27

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-3-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with thiophene-3-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：8.00(dd,J=1.2&3.3Hz,1H),7.45(dd,J=0.9&5.1Hz,1H),7.35(m,2H),7.27(m,2H),6.91(dd,J=9.3&17.1Hz,1H),6.64(ddd,J=2.1,4.8&8.7Hz,1H),6.34(dt,J=5.4,8.7&14.1Hz,1H),5.98(br d,J=2.1Hz,D₂O may be exchanged, 1H).

Example 28

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) furan-2-sulfonamide:

following general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with furan-2-sulfonyl chloride to give the title compound.¹HNMR(300MHz,CDCl₃)：7.53(br s,D₂O exchangeable, 1H),7.38(dd, J = 1.8)&10.5Hz,1H),7.30(d,J=8.4Hz,1H),7.21(d,J=3.0Hz,1H),6.96(dd,J=8.7&16.5Hz,1H),6.87(ddd,J=1.8,5.1&9.0Hz,1H),6.53(dd,J=1.8&3.6Hz,1H),6.44(dt,J=5.1,8.7&13.8Hz,1H),6.22(br s,D₂O may be exchanged, 1H).

Example 29

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -5-methylthiophene-2-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 5-methylthiophene-2-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.34(dd,J=0.9&10.2Hz,1H),7.30(ddd,J=2.1,4.8&9.0Hz,1H),7.25(d,J=3.9Hz,1H),7.07(m,2H),6.65(dd,J=1.2&3.9Hz,1H),5.89(dt,J=2.4,8.7&17.4Hz,1H),5.54(br s,D₂O may be exchanged, 1H),2.46(s, 3H).

Example 30

5-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-2-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 5-chlorothiophene-2-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.38(dd,J=1.5&10.2Hz,1H),7.32(ddd,J=2.1,5.1&9.3Hz,1H),7.25(d,J=3.9Hz,1H),7.10(dd,J=9.0&18.6Hz,3H),6.84(d,J=4.2Hz,1H),5.86(dt,J=1.8,8.7&17.4Hz,1H),5.49(br s,D₂O may be exchanged, 1H).

Example 31

5-bromo-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-2-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 5-bromothiophene-2-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.39-7.29(m,2H),7.20-7.05(m,3H),6.96(d,J=3.6Hz,1H),5.85(dt,J=2.1,9.0&17.4Hz,1H),5.54(br s,1H)。

Example 32

4-bromo-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-3-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 4-bromothiophene-3-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.48(br m,2H),7.39(dd,J=1.8&10.5Hz,1H),7.28(ddd,J=2.4,4.8&9.0Hz,1H),7.17(d,J=8.4Hz,1H),7.02(m,1H),6.02(dt,J=2.4,8.7&17.4Hz,1H),5.68(br s,1H)。

Example 33

4-bromo-5-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-2-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 4-bromo-5-chlorothiophene-2-sulfonyl chloride. ¹H NMR(300MHz,CDCl₃)：7.42-7.34(m,2H),7.25(br m,3H),7.13(dd,J=9.0&17.1Hz,1H),6.02(dt,J=2.4,6.6&17.4Hz,1H),5.52(br s,1H)。

Example 34

3-bromo-5-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-2-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 3-bromo-5-chlorothiophene-2-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.41(dd,J=2.1&10.5Hz,1H),7.35(br m,2H),7.31(dd,J=2.1&4.2Hz,1H),7.19(d,J=8.7Hz,1H),7.08(dd,J=9.0&17.4Hz,1H),6.02(dt,J=2.1,8.4&17.1Hz,1H),5.59(br s,1H)。

Example 35

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2, 5-dimethylthiophene-3-sulfonamide:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 2, 5-dimethylthiophene-3-sulfonyl chloride.¹H NMR(300MHz,CDCl₃)：7.39(dd,J=1.8&10.2Hz,1H),7.24-7.16(br m,2H),7.13(dd,J=9.0&17.4Hz,1H),6.77(d,J=9.6Hz,1H),5.98(dt,J=2.4,8.7&17.4Hz,1H),5.55(br s,1H),2.33(s,6H)。

Example 36

2, 5-dichloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) thiophene-3-sulfonamide:

according to the general procedure B, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzeneReaction of (E) -1, 2-diamine with 2, 5-dichlorothiophene-3-sulfonyl chloride gives the title compound.¹H NMR(300MHz,CDCl₃)：7.41(dd,J=1.5&10.5Hz,1H),7.28-7.20(m,2H),7.08(dd,J=9.0&17.4Hz,2H),6.99(s,1H),6.03(dt,J=2.1,8.7&17.4Hz,1H),5.56(br s,1H)。

Example 37

3- (N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) sulfamoyl) thiophene-2-carboxylic acid methyl ester:

the title compound was obtained according to general procedure B, reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with methyl 3- (chlorosulfonyl) thiophene-2-carboxylate.¹H NMR(300MHz,CDCl₃)：8.58(s,1H),7.43(dd,J=5.1&10.8Hz,2H),7.35(dd,J=1.8&10.2Hz,1H),7.31(ddd,J=2.1,4.2&9.3Hz,1H),7.04(m,2H),5.88(dt,J=2.7,8.7&17.4Hz,1H),5.65(br s,1H),3.85(s,3H)。

Example 38

5- (N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) sulfamoyl) -l-methyl-lH-pyrrole-2-carboxylic acid methyl ester:

The title compound was obtained by reaction of 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 5- (chlorosulfonyl) -l-methyl-1H-pyrrole-2-carboxylic acid methyl ester according to general procedure B.¹H NMR(300MHz,CDCl₃)：7.37(dd,J=1.8&10.5Hz,1H),7.29(m,2H),7.12-6.94(m,4H),5.87(dt,J=1.8,8.4&17.4Hz,1H),5.56(br s,1H),3.65(s,3H),3.75(s,3H)。

Example 39

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -5-methylisoxazole-4-sulfonamide:

following general procedure a, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with the corresponding sulfonyl chloride to give the title compound. Yield: 22 percent. M/z =508[ M-1 ]]^-。

Example 40

3-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) propane-l-sulfonamide:

the desired product was obtained according to general procedure a, reaction of 5, 6-difluoro-N-1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine with 3-chloropropane-l-sulfonyl chloride.¹H NMR(500MHz,CDCl₃)：=7.39-7.38(d,1H),7.35-7.34(m,1H),7.27-7.26(m,1H),7.10-7.0(q,1H),6.63(s,1H,br),6.15-6.11(q,1H),5.60(s,1H,br),3.60-3.56(t,2H),3.22-3.20(m,2H),2.22-2.16(m,2H)。

EXAMPLE 41

N- (2- (4-chloro-2-fluorophenylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide:

see example 1.¹H NMR(300MHz,CDCl₃)0.85-0.95(m,2H),1.05-1.15(ra,2H),2.2-2.4(m,1H),5.8(s,1H),6.3(t,1H),6.6-7.4(m,5H)；m/z=375[M-1]^-。

Example 42

N- (3, 4-difluoro-2- (4-iodo-2-methylphenylamino) phenyl) cyclopropanesulfonamide:

see example 1.¹H NMR(CDCl₃)0.80-1.0(m,2H),1.05-1.20(m,2H),1.55(s,3H),2.4-2.5(m,1H),5.6(s,1H),6.2(dd,1H),6.4(s,1H),7.1(q,1H),7.3-7.4(m,2H),7.5(s,1H)；m/z=463[M-1]^-。

Example 43

N- (2- (4-tert-butyl-2-chlorophenylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide:

see example 1.¹H NMR(300MHz,CDCl₃)0.9-1.0(m,2H),1.05-1.20(m,2H),1.3(s,9H),2.4-2.5(m,1H),5.8(s,1H),6.3(dd,1H),6.6(s,1H),7.0-7.2(m,2H),7.3-7.4(m,2H)；m/z=413[M-l]^-。

Example 44

N- (2- (2, 4-dichlorophenylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide:

see example 1. ¹H NMR(300MHz,CDCl₃)0.9-1.0(m,2H),1.05-1.20(m,2H),2.4-2.5(m,1H),6.0(s,1H),6.3(dd,1H),6.6(s,1H),7.0-7.2(m,2H),7.3-7.4(m,2H)；m/z=392[M-1]^-。

Example 45

3-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-trifluoromethyl) phenylamino) phenyl) propane-1-sulfonamide:

see example 1.¹H NMR(300MHz,CDCl₃)：7.39-7.26(m,2H),7.25(m,1H),7.18(dd,J=9.0&17.7Hz,1H),6.78(br s,D₂O interchangeable, 1H),6.50(t, J =8.1Hz,1H),6.00(br D, D)₂O interchangeable, J =1.5Hz,1H),3.63(t, J = 6.0)&6.3Hz,2H),3.29(t,J=7.2&7.8Hz,2H),2.26 (quintuple, 2H); m/z =445[ M-1 ]]^-。

Example 46

N- (3, 4-difluoro-2- (2-chloro-4-trifluoromethyl) phenylamino) methanesulfonamide:

see example 1.¹H NMR(300MHz,CDCl₃)：7.65(d,J=7.8Hz,1H),7.33(m,2H),7.19(dd,J=9.3&17.4Hz,1H),6.90(br s,D₂O exchangeable, 1H),6.45(dd, J = 1.5)&8.4Hz,1H),6.39(brs,D₂O may be exchanged, 1H),3.02(s, 3H); m/z =399[ M-1]^-。

Example 47

3-chloro-N- (3, 4-difluoro-2- (2-chloro-4-trifluoromethyl) phenylamino) phenyl) propane-1-sulfonamide:

see example 1.¹H NMR(300MHz,CDCl₃)：7.66(d,J=1.5Hz,1H),7.36(m,2H),7.19(dd,J=9.0&17.4Hz,1H),6.91(br s,D₂O exchangeable, 1H),6.50(dd, J = 8.4)&1.5Hz,1H),6.37(s,D₂O interchangeable, 1H),3.62(t, J =6.0Hz,2H),3.29(t, J =7.5&7.8Hz,2H),2.27 (quintuple, 2H); m/z =462[ M-1 ]]^-。

Example 48

3-chloro-N- (3, 4-difluoro-2- (2-bromo-4-trifluoromethyl) phenylamino) phenyl) propane-l-sulfonamide:

see example 1.¹H NMR(300MHz,CDCl₃)：7.82(s,1H),7.38(m,2H),7.20(dd,J=9.0&17.7Hz,1H),6.62(br s,D₂O interchangeable, 1H),6.43(D, J =8.4Hz,1H),6.23(s, D)₂O may be exchanged, 1H),3.65(t, J =6.0Hz,2H),3.30(t, J =7.5Hz,2H),2.28 (quintuple, 2H); m/z =506[ M-1]^-。

Example 49

Cyclopropanesulfonic acid (3,4, 6-trifluoro-2- (2-fluoro-4-iodo-phenylamino) -phenyl) -amide:

step A:(2-fluoro-4-iodo-phenyl) - (2,3, 5-trifluoro-6-nitro-phenyl) -amine:

A stirred solution of 2-fluoro-4-iodoaniline (3.64gm,15.37mmol) in anhydrous THF (100ml) was cooled to-78 deg.C under nitrogen and 1.0M lithium hexamethyldisilazide (LiN (SiMe) was added slowly₃)₂) A solution of "LHMDS" (15.37ml,15.37 mmol). The reaction mixture was stirred continuously at-78 ℃ for an additional hour, and then 2,3,4, 6-tetrafluoronitrobenzene was added. The reaction mixture was allowed to warm to room temperature and stirring was continued for a further 16 hours. Ethyl acetate (200ml) was added to the reaction mixture and washed with water. The organic layer was dried over sodium sulfate and further purified by column chromatography to yield a yellow solid (3.75gm, yield: 59.24%). M-H⁺：410.9。¹H NMR(DMSO,300MHz)：6.85(t,1H)；7.38(d,1H)；7.62(m,2H)；8.78(s,1H)。

And B:3,4, 6-trifluoro-N ² - (2-fluoro-4-iodo-phenyl) -benzene-1, 2-diamine:

to a stirred solution of (2-fluoro-4-iodo-phenyl) - (2,3, 5-trifluoro-6-nitro-phenyl) -amine 3(5.2gm,12.62mmol) in EtOH (200ml) was added ammonium chloride (10.12gm,189.3mmol) and iron powder (10.57gm,189.3 mmol). The reaction mixture was refluxed with constant stirring for 16 hours. The reaction mixture was cooled, filtered through celite (celite) and concentrated to dryness. The resulting residue was taken up in EtOAc and washed with water. The EtOAc layer was dried over sodium sulfate and further purified by crystallization from EtOH to yield an off-white solid (3.2gm, yield: 66.39%). M-H ⁺：381.1。¹H NMR(DMSO,300MHz)：5.0(s,2H)；6.2(t,1H)；7,2-7.3(m,2H)；7.45(s,1H)；7.5(d,1H)。

And C:4,6, 7-trifluoro-l- (' 2-fluoro-4-iodo-phenyl) -1, 3-dihydrobenzimidazol-2-one：

To 3,4, 6-trifluoro-N2- (2-fluoro-4-iodo-phenyl) -benzene-1, 2-diamine 3(0.285gm,0.74mmol) in CH₂Cl₂To the stirred solution in (2ml) was added 1, 1' -carbonyldiimidazole (0.125gm,0.75 mmol). The reaction mixture was stirred continuously at room temperature for 16 hours, during which time the product precipitated out. The white solid was filtered and used further without any purification (0.2gm, yield: 65.85%). M/z =407[ M-1%]^-。

Step D/E:cyclopropanesulfonic acid (3,4, 6-trifluoro-2- (2-fluoro-4-iodo-phenylamino) -phenyl) -amide:

a stirred solution of 4,6, 7-trifluoro-l- (2-fluoro-4-iodo-phenyl) -1,3, -dihydrobenzimidazol-2-one (0.2gm,0.41mmol) in anhydrous THF (4ml) was cooled to-78 ℃ under nitrogen and 1.0MLiHMDS solution (0.41ml,0.41mmol) was added slowly. (2ml) Cyclopropanesulfonyl chloride (0.050ml,0.49mmol) was then added. The reaction mixture was stirred continuously at room temperature for 16 h, concentrated to dryness and placed in EtOAc. The EtOAc portion was washed with water, dried over sodium sulfate, and concentrated to dryness. The resulting residue, l-cyclopropanesulfonyl-4, 5, 7-trifluoro-3- (2-fluoro-4-iodo-phenyl) -1, 3-dihydro-benzimidazol-2-one 5, was taken up in dioxane (2ml) and 1.0N NaOH (0.5ml) was added thereto and stirring was continued at 50 ℃ for 16 hours. TLC indicated incomplete reaction and the product was purified by HPLC to give an off-white solid (4.4 mg). M + H ⁺：484.7,M-H⁺：486.7。¹HNMR(CDCl₃,300MHZ)：0.9-1.1-(m,2H)；1.1-1.2(m,2H)；2.45-2.55(m,1H)；6.05(s,1H)；6.44-6.54(m,1H)；7.1(s,1H)；7.4-7.7(d,1H)；7.38-7.44(dd,1H)；m/z=485[M-1]^-。

Example 50

N- (3, 4-difluoro-2- (4-fluoro-2-iodophenylamino) -6-ethoxyphenyl) cyclopropanesulfonamide:

step A:(2, 3-difluoro-5-methoxy-6-nitro-phenyl) - (2-fluoro-4-iodo-phenyl) -amine:

a stirred solution of (2-fluoro-4-iodo-phenyl) - (2,3, 5-trifluoro-6-nitro-phenyl) -amine (1.23gm,3mmol) in anhydrous THF (25ml) was cooled to-78 ℃ under nitrogen and a 25% NaOMe solution (0.68ml,0.3mmol) was slowly added. The reaction mixture was allowed to warm to room temperature and stirring was continued for a further 16 hours. TLC indicated incomplete reaction. Ethyl acetate (100ml) was added to the reaction mixture and washed with water. The organic layer was dried over sodium sulfate and further purified by column chromatography to yield a yellow solid (0.6gm, yield: 47.6%). M/z =424[ M = H%]⁺。

And B:5, 6-difluoro-N1- (4-fluoro-2-iodophenyl) -3-methoxybenzene-1, 2-diamine:

to a stirred solution of (2, 3-difluoro-5-methoxy-6-nitro-phenyl) - (2-fluoro-4-iodo-phenyl) -amine (0.57gm,1.34mmol) in EtOH (20ml) were added ammonium chloride (1.18gm,20.16mmol) and iron powder (1.15gm,21.44 mmol). The reaction mixture was refluxed with constant stirring for 16 hours. The reaction mixture was cooled, filtered through celite, and concentrated to dryness. The resulting residue was taken up in EtOAc and washed with water. The EtOAc layer was dried over sodium sulfate and further purified by crystallization from EtOH to yield an off-white solid (0.47gm, yield: 90.3%). M-H ⁺：393.2。¹H NMR(DMSO,300MHz)：3.76(s,3H)；6.1(t,1H)；6.8-7.0(m,1H)；7.2(d,1H)；7.35(s,1H)；7.42(d,1H)。

And C:6, 7-difluoro-l- (4-fluoro-2-iodophenyl) -4-methoxy-lH-benzo [ d]Imidazol-2 (3H) -one:

to a solution of 5, 6-difluoro-N1- (4-fluoro-2-iodophenyl) -3-methoxybenzene-1, 2-diamine (0.17gm,0.43mmol) in CH₂Cl₂To a stirred solution in (2ml) was added 1, 1' -carbonyldiimidazole (0.085gm,0.53 mmol). The reaction mixture was stirred continuously at room temperature for 16 hours, during which time the product precipitated out. The white solid was filtered and used further without any purification (0.089 gm); m/z =419[ M-1 ]]^-。

Step D/F:n- (3, 4-difluoro-2- (4-fluoro-2-iodophenylamino) -6-methoxyphenyl) cyclopropanesulfonamide:

under nitrogen, 1- (cyclopropylsulfonyl) -4, 5-difluoro-3- (2-fluoro-4-iodophenyl) -7-methoxy-lH-benzo [ d]A stirred solution of imidazol-2 (3H) -one (0.89gm,0.17mmol) in dry THF (4ml) was cooled to-78 deg.C and a 1.0M solution of LiHMDS (0.17ml,0.17mmol) was added slowly. (2ml) Cyclopropanesulfonyl chloride (0.021ml,0.21mmol) was then added. The reaction mixture was stirred continuously at room temperature for 16 h, concentrated to dryness and placed in EtOAc. The EtOAc fractions were washed with water, dried over sodium sulfate, and concentrated to dryness. The resulting l- (cyclopropylsulfonyl) -4, 5-difluoro-3- (2-fluoro-4-iodophenyl) -7-methoxy-lH-benzo [ d ]Imidazol-2 (3H) -one was placed in dioxane (2ml) and 1.0N NaOH (0.5ml) was added thereto and stirring was continued at 50 ℃ for 16 hours. TLC indicated incomplete reaction and the product was purified by HPLC to give an off-white solid (2.5 mg). M + H⁺：484.7,M-H⁺：497.3。¹H NMR(CDCl₃,300MHz)：0.85-0.95(m,2H)；1.05-1.15(m,2H)；2.4-2.5(m,1H)；3.9(s,3H)；6.1(s,1H)；6.4-6.6(m,2H)；7.3(m,1H)；7.35-7.4(dd,1H)；m/z=497[M-1]^-。

Example 51

Methanesulfonic acid (3, 4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -6-methoxy-phenyl) -amide:

to 5, 6-difluoro-N1- (4-fluoro-2-iodophenyl) -3-methoxybenzene-1, 2-diamine (0.150gm,0.38mmol) in anhydrous CH₂Cl₂To the stirred solution in (4ml) was added slowly TEA (264ml,1.9mmol) and methanesulfonyl chloride. The reaction mixture was stirred continuously at room temperature for 16 hours, TLC showed incomplete reaction and starting material and two products were observed. The reaction mixture was washed with water, the organic layer was dried over sodium sulfate and concentrated to dryness, and the product was purified by column chromatography. The minor product was found to be the expected compound (6.4 mg). M-H⁺：471.5。¹H NMR(CDCl₃,300MHz)：3.9(s,3H)；6.05(s,1H)；6.4-6.5(m,1H)；6.5-6.6(m,1H)；7.2(s,1H)；7.28(d,1H)；7.35-7.4(d,1H)；m/z=471[M-1]^-。

Example 52

l- (2, 3-dihydroxy-propyl) -cyclopropanesulfonic acid [3,4, 6-trifluoro-2- (4-fluoro-2-iodo-phenylamino) -phenyl ] -amide:

step A:1-allyl-cyclopropanesulfonic acid [3,4, 6-trifluoro-2- (2-fluoro-4-iodo-phenylamino) phenyl]-acyl Amine:

according to the general procedure B, 1-allyl-cyclopropanesulfonyl chloride is reacted with 3,5, 6-trifluoro-N¹- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine to give the title product. ¹H NMR(CDCl₃,300MHz)：7.41(dd,1H),7.38(dd,1H),7.09(s,1H),6.78(m,1H),6.49(m,1H),5.96(s,1H),5.86(m,1H),5.18(d,2H),2.76(d,2H),1.23(m,2H),0.872(m,2H)。

And B:l- (2, 3-dihydroxypropyl) -N- (3,4, 6-trifluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane Alkane-1-sulfonamides:

1-allyl-cyclopropanesulfonic acid [3,4, 6-trifluoro-2- (2-fluoro-4-iodo-phenylamino) -phenyl]The amide (110mg,0.21mmol) and 4-methylmorpholine N-oxide (24.6mg,0.21mmol) were dissolved in THF (8 mL). Osmium tetroxide (0.021mmol,0.153mL,4% in H) was added at room temperature₂O) and the reaction mixture was stirred at room temperature for 16 hours. EtOAc was added and the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product (0.89g, 75%).¹H NMR(CDCl₃,300MHz)：7.39(dd,J=1.5&10.6Hz,1H),7.29(d,J=8.8Hz,IH),7.28(s,1H),6.97(s,1H),6.76(m,1H),6.49(m,1H),4.13(m,1H),3.66(dd,J=3.7&11.4Hz,1H),3.53(dd.J=6.7&11.2Hz,1H),2.50(dd,J=10.0&16.1Hz,1H),1.6(m,lH),1.46(m,1H),1.28(m,1H),1.20(m,2H),0.92(m,2H)；m/z=559[M-1]^-。

Example 53

(S) -l- (2, 3-dihydroxypropyl) -N- (3,4, 6-trifluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-l-sulfonamide:

the racemic mixture (example 52) was separated by chiral HPLC to give the pure S isomer.¹H NMR(CDCl₃,300MHz)：7.39(dd,J=1.5&10.6Hz51H),7.29(d,J=8.8Hz,1H),7.28(s,1H),6.97(s,1H),6.76(m,1H),6.49(m,1H),4.13(m,1H),3.66(dd,J=3.7&11.4Hz51H),3.53(dd,J=6.7&11.2Hz,1H),2.50(dd,J=10.0&16.1Hz,1H),1.6(m,lH),1.46(m,1H),1.28(m,1H),1.20(m,2H),0.92(m,2H)；m/z=559[M-1]^-。

Example 54

(R) -l- (2, 3-dihydroxypropyl) -N- (3,4, 6-trifluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-l-sulfonamide:

the racemic mixture (example 52) was separated by chiral HPLC to give the pure R isomer.¹H NMR(CDCl₃,300MHz)：7.39(dd,J=1.5&10.6Hz,1H),7.29(d,J=8.8Hz,1H),7.28(s,1H),6.97(s,1H),6.76(m,1H),6.49(m,1H),4.13(m,1H),3.66(dd,J=3.7&11.4Hz,1H),3.53(dd,J=6.7&11.2Hz,1H),2.50(dd,J=10.0&16.1Hz,1H),1.6(m,lH),1.46(m,1H),1.28(m,1H),1.20(m,2H),0.92(m,2H)；m/z=559[M-1]^-。

Example 55

Step A:1-allyl-N-3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane- 1-sulfonamides：

Following general procedure B, 1-allyl-cyclopropanesulfonyl chloride was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methoxybenzene-1, 2-diamine to give the title product.¹H NMR(CDCl₃,300MHz)：7.417(dd,1H),7.309(s,1H),7.25(m,1H),6.89(m,1H),6.52(m,1H),6.427(m,1H),6.03(s,lH),5.668(m,1H),5.11(t,1H),3.9(s,3H),2.75(d,2H),1.21(m,2H),0.767(m,2H)。

And B:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1- (2, 3-dihydroxypropyl) cyclopropyl Alkane-1-sulfonamides：

l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-l-sulfonamide (97mg,0.18mmol) and 4-methylmorpholine N-oxide (21mg,0.18mmol) were dissolved in THF (8 mL). Osmium tetroxide (0.018mmol,0.13mL,4% in H) was added at room temperature₂O) and the reaction mixture was stirred at room temperature for 16 hours. EtOAc was added and the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product (0.80g, 78%).¹H NMR(CDCl₃,300MHz)：7.38(dd,J=1.7&10.3Hz,1H),7.26(m,1H),7.14(s,1H),6.87(s,1H),6.53(dd,J=6.8&11.4Hz,1H),6.43(m,1H),4.06(m,1H),3.89(s,3H),3.63(dd,J=3.7&11.1Hz,1H),3.49(dd,J=6.4&11.1Hz,1H),2.3(dd,J=9.7&16.1Hz,1H),1.77(dd,J=1.9&16.0Hz,1H),1.37(m,1H),1.25(m,1H),1.21(m,2H),0.86(m,2H)；m/z=571[M-1]^-。

Example 56

(S) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide:

the racemic mixture (example 55) was separated by chiral HPLC to give the pure S isomer.¹H NMR(CDCl₃,300MHz)：7.38(dd,J=1.7&10.3Hz,1H),7.26(m,1H),7.14(s,1H),6.87(s,1H),6.53(dd,J=6.8&11.4Hz,1H),6.43(m,1H),4.06(m,1H),3.89(s,3H),3.63(dd,J=3.7&11.1Hz,1H),3.49(dd,J=6.4&11.1Hz,1H),2.3(dd,J=9.7&16.1Hz,1H),1.77(dd,J=1.9&16.0Hz,1H),1.37(m,1H),1.25(m,1H),1.21(m,2H),0.86(m,2H)；m/z=571[M-1]^-。

Example 57

(R) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-l-sulfonamide:

The racemic mixture (example 55) was separated by chiral HPLC to give the pure R isomer.¹H NMR(CDCl₃,300MHz)：7.38(dd,J=1.7&10.3Hz,1H),7.26(m,1H),7.14(s,1H),6.87(s,1H),6.53(dd,J=6.8&11.4Hz,1H),6.43(m,1H),4.06(m,1H),3.89(s,3H),3.63(dd,J=3.7&11.1Hz,1H),3.49(dd,J=6.4&11.1Hz,1H),2.3(dd,J=9.7&16.1Hz,1H),1.77(dd,J=1.9&16.0Hz,1H),1.37(m,1H),1.25(m,1H),1.21(m,2H),0.86(m,2H)；m/z=571[M-1]^-。

Example 58

1- (2-hydroxyethyl) -N- (3,4, 6-trifluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-1-sulfonamide:

step A:TBS protected 1-, (2-hydroxyethyl) -N- (3,4, 6-trifluoro-2- (2-fluoro-4-iodophenylamino) benzene Yl) cyclopropane-1-sulfonamides：

Following general procedure B, the sulfonyl chloride prepared in step C of example 16 was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-fluorobenzene-1, 2-diamine to give the title product. Yield: 13 percent.¹H-NMR(300MHz,CDCl₃)：=7.51(s,1H,br),7.37-7.35(d,1H),7.27-7.25(d,1H),6.94(s,1H,br),6.78-6.68(m,1H),6.46-6.44(m,1H),3.90-3.88(t,2H),2.12-2.10(t,2H),1.31-1.28(m,2H),0.91-0.89(m,2H),0.86(s,9H),0.05(s,6H)；m/z=643[M-l]^-。

And B:1- (2-hydroxyethyl) -N- (3,4, 6-trifluoro-2- (2-fluoro-4-iodophenylamino) -phenyl) cyclopropane- 1-sulfonamides：

The same procedure as in step E of example 16. Yield: 100 percent.¹H-NMR(300MHz,CDCl₃)：=7.51(s,1H,br),7.37-7.35(d,1H),7.27-7.25(d,1H),6.94(s,1H,br),6.78-6.68(m,1H),6.46-6.44(m,1H),3.90-3.88(t,2H),2.12-2.10(t,2H),1.31-1.28(m,2H),0.91-0.89(m,2H)；m/z=529[M-1]^-。

Example 59

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2-hydroxyethyl) cyclopropane-1-sulfonamide:

step A:TBS protected N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2- Hydroxyethyl) cyclopropane-1-sulfonamides：

Following general procedure B, the sulfonyl chloride prepared in step C of example 16 was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methoxy-benzene-1, 2-diamine to give the title product. Yield: 37 percent. ¹H-NMR(300MHz,CDCl₃)：=7.40-7.34(dd,1H),7.23-7.21(m,1H),6.61(s,1H,br),6.57-6.49(dd,1H),6.48-6.39(m,1H),3.9-3.7(m,5H),2.15-2.05(t,2H),1.30-1.20(m,2H),0.95-0.80(m,11H),0.05(s,6H)；m/z=655[M-1]^-。

And B:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2-hydroxyethyl) Cyclopropane-1-sulfonamides：

The same procedure as in step E of example 16. Yield: 100 percent.¹H-NMR(300MHz,CDCl₃)：=7.40-7.34(dd,1H),7.23-7.21(m,1H),6.61(s,1H,br),6.57-6.49(dd,1H),6.48-6.39(m,1H),3.9-3.7(m,5H),2.15-2.05(t,2H),1.30-1.20(m,2H),0.95-0.80(m,2H)；m/z=541[M-1]^-。

Example 60

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (3-hydroxy-2- (hydroxymethyl) propyl) cyclopropane-1-sulfonamide:

step A:2- (2-bromoallyl) malonic acid dimethyl ester：

To a solution of sodium hydride (5.0g,125mmol) in HMPA (50ml, hydrogen fromCalcium chloride distillation) was added a solution of dimethyl malonate (11.7ml,100mmol) in HMPA (5 ml). The mixture was heated to 50 ℃ and stirred for 1 hour. After this time, the solution was cooled to 0 ℃ again and a solution of 2, 3-dibromopropene (12.2ml,100mmol) in HMPA (5ml) was added to the reaction mixture. The solution was then allowed to warm to 40 ℃ and stirred for 1 hour. The reaction mixture was quenched with aqueous HCl (10%,88ml) and extracted with ether (3 × 45 ml). The organic fraction was collected over MgSO₄Dried and the solvent removed in vacuo. The crude oil was purified by silica gel chromatography (eluent: chloroform/hexane) to give the title product as a colorless oil (16.3g, 65%).¹H-NMR(300MHz,CDCl₃)5.70(d,J=1.8Hz,1H),5.48(d,J=1.8Hz,1H),3.63(t,J=7.5Hz,1H),3.76(s,6H),3.04(d,J=7.5Hz,2H)。

And B:2- (2-bromoallyl) propane-1, 3-diol ：

Lithium aluminum hydride (1.9g,7.65mmol) was slurried in anhydrous diethyl ether (50ml) and cooled to-78 ℃ in a dry ice/acetone bath. A solution of the product from step A (0.639g,16.84mmol) in dry ether (26ml) was then added dropwise. After addition of this malonate, the solution was allowed to warm to room temperature and stirring was continued for 3 hours. The reaction was quenched with brine (50ml), extracted with ethyl acetate (3 × 25ml), and over MgSO₄And (5) drying. The solvent was removed in vacuo to yield the desired product (1.3g,86%) which was used in the next step without further purification.¹H-NMR(300MHz,CDCl₃)5.66(d,J=1.2Hz,1H),5.48(d,J=1.5Hz,1H),3.86(m,2H),3.73(m,2H),2.51(d,J=7.5Hz,2H),2.40(br s,2H),2.15(m,1H)。

And C:di-tert-butyldimethylsilyl-protected 2- (2-bromoallyl) propane-1, 3-diol：

The product from step B (2.8g,14.20mmol) was dissolved in dry THF (140 ml). Anhydrous pyridine (2.5ml,31.24mmol) was added and the solution was cooled to 0 ℃. Tert-butyldimethylsilyltrifluoromethanesulfonate (7.2ml,31.24mmol) was added dropwise and when the addition was complete, the reaction solution was heated to 35 ℃. After stirring for 6 days, the reaction was quenched with 100ml brine, extracted with ethyl acetate (3 × 50ml) and over MgSO₄And (5) drying. The combined organic phases were evaporated to give the crude product as a yellow oil (5.5g,91%) which was used in the next step without further purification. ¹H-NMR(300MHz,CDCl₃)5.54(d,J=0.9Hz,1H),5.40(d,J=1.2Hz,1H),3.55(d,J=5.4,4H),2.40(d,J=6.9Hz,2H),1.97(m,1H),0.85(s,18H),0.02(s,9H)。

Step D:di-tert-butyldimethylsilyl-protected 2- ((1-bromocyclopropyl) methyl) propane-1, 3-diol：

Anhydrous CH was added to the reaction flask at 0 deg.C₂Cl₂(10ml) and diethyl zinc (1.0M in hexanes, 4.65ml,4.65 mmol). Trifluoroacetic acid (0.358ml,4.65mmol) was added dropwise and the solution was stirred for 20 min. Diiodomethane (0.375ml,4.65mmol) was then added and the solution stirred for an additional 20 minutes. Finally, the product from step C (0.492g,1.16mmol) was added and the solution was allowed to warm to ambient temperature and stirred for 16 h. With saturated NH₄The reaction was terminated with aqueous Cl solution. The layers were partitioned and the aqueous phase was extracted with chloroform (3x5 ml). The combined organic phases were washed with brine (10ml) over MgSO₄Dried and the volatiles removed in vacuo. The crude product obtained was purified by silica gel chromatography (eluent: chloroform/hexane) to give the product as a clear oil (0.280g, 64%).¹H-NMR(300MHz,CDCl₃)3.66(d,J=5.4,4H),2.08(m,1H),1.64(d,J=6.9,2H),1.13(m,2H),0.88(s,18H),0.81(m,2H),0.04(s,9H)。

Step E:di-tertButyldimethylsilyl-protected 1- (3-hydroxy-2- (hydroxymethyl) propyl) cyclopropane- 1-sulfonyl chlorides：

The product from step D (0.507g,1.16mmol) was dissolved in dry ether (6ml) and the reaction solution was cooled to-78 ℃. Thereafter, tert-butyllithium (1.7M in pentane, 1.50ml,2.55mmol) was added dropwise over 5 minutes. After stirring for 0.5 h, the lithiated product was transferred by cannula to a stirred solution of sulfonyl chloride (0.206ml,2.55mmol) in dry ether (6ml) at-78 ℃. Once the transfer was complete, the solution was allowed to warm to room temperature, the solvent was evaporated, and the resulting white solid was slurried in anhydrous hexane. The slurry was immediately filtered through celite and all volatiles were removed in vacuo. The resulting crude product was isolated as a yellow oil (0.376g,71%) and used in the next step without further purification. ¹H-NMR(300MHz,CDCl₃)3.60(m,4H),2.16(m,1H),2.03(d,2H),0.88(s,18H),0.04(s,9H)。

Step F:di-tert-butyldimethylsilyl-protected N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) - 6-methoxyphenyl) -1- (3-hydroxy-2- (hydroxymethyl) propyl) cyclopropane-1-sulfonamide：

5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methoxybenzene-1, 2-diamine (8.8mg,0.022mmol) was dissolved in anhydrous pyridine (0.5ml) under an argon atmosphere. The product from step E (20.5mg,0.045mmol) dissolved in anhydrous pyridine (0.5ml) was added to the reaction flask and the mixture was added at 80 ℃ for 21 h. The solvent was removed in vacuo, and the resulting crude product was purified by silica gel chromatography (eluent: ethyl acetate/hexane) to give the title compound (2.75mg, 15%). M/z813.5 (M-1).

Step G:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (3-hydroxy-2- (hydroxy) Ylmethyl) propyl) cyclopropane-1-sulfonamide：

The product from step F (27.9mg,0.0342mmol) was dissolved in THF (1ml) and treated with aqueous HCl (1.2N,0.2ml) at 0 ℃. The resulting solution was stirred for 4 hours. Thereafter, saturated NaHCO was used₃The reaction was quenched with aqueous ethyl acetate, extracted over MgSO₄Dried and the volatiles removed in vacuo. The resulting crude product was purified by silica gel chromatography (eluent: methanol/chloroform) followed by LC-MS to yield the title compound (11.8mg, 59%). ¹H-NMR(300MHz,CD₃OD)7.32(dd,1H),7.21(d,1H),6.76(dd,1H),6.33(m,1H),3.82(s,3H),3.52(d,4H),2.01(m,1H),1.88(d,2H),1.07(m,2H),0.75(m,2H),m/z585.3(M-1)^-。

Example 61

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclobutanesulfonamide:

step A:cyclobutanesulfonyl chloride：

To a suspension of Mg turnings (0.790g,32.5mmol) in 20ml of anhydrous diethyl ether was added a solution of cyclobutylbromide (1.8ml,2.5722g,19.1mmol) in 20ml of diethyl ether, in small portions, with vigorous stirring. After the initial exothermic reaction had ceased, the mixture was heated to reflux temperature for 30 min. The suspension was cooled to room temperature and the supernatant was added portionwise to an ice-cold solution of sulfonyl chloride (4.6ml,7.728g,57.2mmol) in 30ml of anhydrous DCM. After the addition was complete, the suspension was warmed to room temperature and the volatiles were removed in vacuo. The residue was dried in a vacuum oil pump for 15min and then extracted with hexane (150 ml). The hexane suspension was filtered and the hexane removed in vacuo to yield the crude product as a dark purple oil which was used in the next step without further purification. Some unreacted cyclopropyl bromide is still present. Yield of crude product: 1.1g (38%).

And B:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclobutanesulfonamide：

Following general procedure B, the cyclobutyl sulfonyl chloride prepared in the above step was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methoxybenzene-1, 2-diamine to give the title product. Yield: 75 percent. ¹H-NMR(300MHz,CDCl₃)：=7.44(s,1H,br),7.41-7.36(dd,1H),7.24-7.23(m,1H),6.54-6.38(m,2H),5.90(s,1H,br),3.85-3.75(m,5H),2.60-2.40(m,2H),2.25-2.15(m,1H),2.15-1.95(m,2H)；m/z=511[M-1]^-。

Example 62

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methylphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide:

step A:(3,4, 5-trifluorophenyl) methanol：

To a cooled (-5 ℃ C.) solution of 3,4, 5-trifluorobenzaldehyde (7.0g,43.75mmol) in a mixture of THF and water (50ml,9:1) was slowly added NaBH in portions over 30min₄(1.662g,43.75 mmol). In 2The reaction mixture was allowed to reach room temperature over a period of h and carefully poured into ice-cold dilute HCl (200ml, 1N). Extracting the oil layer into CH₂Cl₂In (250ml), the organic layer was washed with water (200ml) and dried (MgSO)₄) And evaporated. The resulting crude product (7.08g, quantitative) was used without further purification.

And B:5- (bromomethyl) -1,2, 3-trifluorobenzene：

To (3,4, 5-trifluorophenyl) methanol (40mmol) in CH₂Cl₂To a solution in (150ml) thionyl bromide (6.16ml,80mmol) in CH was added slowly₂Cl₂(50 ml). The reaction mixture was stirred at room temperature for 16h and poured into ice water (200 ml). The organic layer was separated and washed with saturated NaHCO₃(2X200ml), water (200ml) and dried (MgSO₄) And evaporated to give the corresponding bromo compound as a pale yellow oil in quantitative yield. The crude product was used for the next reaction without further purification.

And C:1,2, 3-trifluoro-5-methylbenzene：

The above bromine compound (40mmol) was mixed with triethylsilane (48mmol), and the mixture was treated with solid PdCl₂(4mmol) the reaction mixture was worked up in small portions. After a few minutes, a violent exothermic reaction ensued, taking care to reflux the contents of the flask by installing a reflux condenser. The reaction mixture was stirred at room temperature for a further 6h and the contents were allowed to stand for 16 h. The crude liquid product was then carefully poured off and used on for the next reaction without further purification. The reaction is assumed to proceed in quantitative yield.

Step D:1,2, 3-trifluoro-5-methyl-4-nitrobenzene：

1,2, 3-trifluoro-5-methylbenzene (40mmol) was added to concentrated H at 0-5 deg.C₂SO₄(50 ml). Then using concentrated HNO₃(3.39ml,48.44mmol,90%) the reaction mixture was slowly worked up while maintaining the internal temperature below 20 ℃. The reaction mixture was stirred at room temperature for 16h, poured onto ice (300g) and washed with CH₂Cl₂(2x125ml) extract the oil layer. The organic layer was washed with water (2X200ml), brine (200ml) and dried (MgSO)₄) And evaporated to give the crude product which was purified by flash silica gel chromatography to give the title product (6.5g, 85%).¹H-NMR(300MHz,CDCl₃): 6.96 (heptad, 1H),2.39(s, 3H). ¹⁹FNMR(CDCl₃)：-128.18,-141.50,-159.05。

Step E:2, 3-difluoro-N- (2-fluoro-4-iodophenyl) -5-methyl-6-nitroaniline：

The title compound was formed by reacting 2-fluoro-4-iodoaniline with 1,2, 3-trifluoro-5-methyl-4-nitrobenzene using the conditions described in example 1 (step a). M-H⁺：407.9。

Step F:5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methylbenzene-1, 2-diamine：

The title compound was formed by reduction of 2, 3-difluoro-N- (2-fluoro-4-iodophenyl) -5-methyl-6-nitroaniline using the conditions described in example 1 (step B). M-H⁺：377.4。

Step G:l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methylphenyl) cyclopropane-1- Sulfonamides：

Following general procedure B, 1-allyl-cyclopropanesulfonyl chloride (142mg ) was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methylbenzene-1, 2-diamine (150mg,0.4mmol) to give the title product (100mg, 47%); m/z =521[ M-1]^-。

Step H:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methylphenyl) -1- (2, 3-dihydroxypropane Yl) cyclopropane-1-sulfonamides：

1-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methylphenyl) cyclopropane-1-sulfonamide (150mg,0.29mmol) and 4-methylmorpholine N-oxide (33mg,0.29mmol) were dissolved in THF (5 mL). Osmium tetroxide (0.029mmol,0.18mL,4% in H) was added at room temperature ₂O) and the reaction mixture was stirred at room temperature for 16 hours. EtOAc was added and the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product (0.110g, 68%).¹H-NMR(300MHz,CDCl₃)：7.07(m,1H),6.97(br m,2H),6.84(m,2H),6.60(br m,2H),3.98(br m,1H),3.58(m,1H),3.43(m,1H),3.20(d,J=3.9Hz,1H),2.42(s,3H),2.31(dd,J=9.9&15.6Hz,1H),2.01(br t,1H),2.31(dd,J=9.9&15.6Hz,1H),1.66(dd,J=2.1&15.9Hz,1H),1.52(m,1H),1.40(m,1H),0.91(m,2H)。

Example 63

1- (2, 3-dihydroxypropyl) -N- (6-ethyl-3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-1-sulfonamide:

step A:1- (3,4, 5-trifluorophenyl) ethanol：

A solution of MeMgBr in ether (17.41ml,52.24mmol,3M) was added slowly to a solution of 3,4, 5-trifluorobenzaldehyde (6.96g,43.53mmol) in THF (125ml) at-78 ℃. The reaction mixture was stirred at room temperature for 16h, cooled (0 ℃ C.), and quenched with excess ethyl acetate (10ml) and water (5ml) in that order. Adding excess anhydrous MgSO₄(5g) And stirred at room temperature for 30 minutes. The suspension was filtered through celite and the solid was washed with ethyl acetate (2 × 25 ml). The combined filtrates were evaporated to give the product (7.65g) in quantitative yield.

And B:5- (l-bromoethyl) -1,2, 3-trifluorobenzene：

To a solution of 1- (3,4, 5-trifluorophenyl) ethanol (7.65g,43.5mmol) in CH₂Cl₂To a solution in (250ml) thionyl bromide (18.1g,87mmol) in CH was added slowly₂Cl₂(50 ml). The reaction mixture was stirred at room temperature for 16h and poured into ice water (200 ml). The organic layer was separated and washed with saturated NaHCO ₃(2X200ml), water (200ml) and dried (MgSO₄) And evaporated to give the corresponding bromine compound (10.4g) as a pale yellow oil in quantitative yield. The crude product was used for the next reaction without further purification.

And C:5-ethyl-1, 2, 3-trifluorobenzene：

The above bromine compound (9.65g,40.4mmol) was mixed with triethylsilane (41mmol), and the mixture was treated with solid PdCl₂(177mg,4mmol) the reaction mixture was worked up in small portions. After a few minutes, a violent exothermic reaction ensued, taking care to reflux the contents of the flask by installing a reflux condenser. The reaction mixture was stirred at room temperature for a further 6h and the contents were allowed to stand for 16 h. The crude liquid product was then carefully poured off and used on for the next reaction without further purification. The reaction is assumed to proceed in quantitative yield.

Step D:1-ethyl-3, 4, 5-trifluoro-2-nitrobenzene：

1,2, 3-trifluoro-5-methylbenzene (6.46g,40.4mmol) was added to concentrated H at 0-5 deg.C₂SO₄(50 ml). Then using concentrated HNO₃(3.39ml,48.44mmol,90%) the reaction mixture was slowly worked up while maintaining the internal temperature below 20 ℃. The reaction mixture was stirred at room temperature for 16h, poured onto ice (300g) and washed with CH ₂Cl₂(2x125ml) extract the oil layer. The organic layer was washed with water (2X200ml), brine (200ml) and dried (MgSO)₄) And evaporated to give the crude product which was purified by flash silica gel chromatography to give the title product (6.6g, 79%).¹H NMR(CDCl₃): 6.98 (heptad, 1H),2.68(q,2H),1.26(t, J = 7.8)&7.2Hz,3H)。

Step E:3-ethyl-5, 6-difluoro-N- (2-fluoro-4-iodophenyl) -2-nitroaniline：

Using the conditions described in example 1 (step a),reacting 2-fluoro-4-iodoaniline (2.05g,10mmol) and 1-ethyl-3, 4, 5-trifluoro-2-nitrobenzene (2.37g,10mmol) to form the title compound (2.47g, 60%); m/z =407[ M-1%]^-。

Step F:3-Ethyl-5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine：

1,2, 3-trifluoro-5-methyl-4-nitrobenzene (2.47g,5.85mmol) was reduced using the conditions described in example 1 (step B) to form the title compound. M-H⁺：393。

Step G:l-allyl-N- (6-ethyl-3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropane-l- Sulfonamides：

Following general procedure B, l-allyl-cyclopropanesulfonyl chloride (230mg,1.27mmol) was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3-methylbenzene-1, 2-diamine (100mg,0.255mmol) to give the title product (72mg, 53%); m/z =535[ M-1 ].

Step H:1- (2, 3-dihydroxypropyl) -N- (6-ethyl-3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) benzene Yl) cyclopropane-l-sulfonamides：

l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methylphenyl) cyclopropane-1-sulfonamide (70mg,0.13mmol) and 4-methylmorpholine N-oxide (15mg,0.13mmol) were dissolved in THF (2 mL). Osmium tetroxide (0.013mmol,0.075mL,4% in H) was added at room temperature₂O) and the reaction mixture was stirred at room temperature for 16 hours. Adding EtOAc, the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product.¹H NMR(300MHz,CDC1₃)：7.38(dd,J=2.1&10.8Hz,1H),7.27(m,2H),7.12(br s,1H),6.91(dd,J=8.1&10.8Hz,1H),6.69(br s,1H),6.36(dt,J=4.8,8.7&13.5Hz,1H),4.00(m,1H),3.62(dd,J=3.6&10.5Hz,1H),3.47(br m,2H),2.81(q,2H),2.40(dd,J=10.2&15.9Hz,1H),1.73(br m,2H),1.58(m,1H),1.43(m,1H),0.94(m,2H)。

Example 64

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- (2-methoxyethoxy) phenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide:

step A:1,2, 3-trifluoro-5- (2-methoxyethoxy) -4-nitrobenzene：

To 3,4, 5-trifluoro-2-nitrophenol (1.93,10mmol), Ph at 0 deg.C₃To a mixture of P (3.93g,15mmol) and 2-methoxy-ethanol (1.18ml,15mmol) in dry THF (25ml) was added a solution of diisopropyl azodicarboxylate (2.91ml,15mmol) in THF (5ml) and the reaction mixture was stirred at room temperature for 16 h. Evaporate the volatiles and dissolve the residue in CH₂Cl₂(100ml), the organic layer was washed with water (100ml), brine (100ml) and dried (MgSO ₄) And evaporated. The residue obtained was purified by flash chromatography on silica gel to give the title product in a yield of 68% (1.70 g).¹HNMR(300MHz,CDCl₃)：6.78(ddd,J=2.4,6.0,11.7Hz,1H),4.19(t,J=4.5Hz,2H),3.72(t,J=4.5Hz,2H),3.39(s,3H)。

And B:2, 3-difluoro-N- (2-fluoro-4-iodophenyl) -5- (2-methoxyethoxy) -6-nitroaniline：

Using the conditions described in example 1 (step a), 2-fluoro-4-iodoaniline (1.6g,6.8mmol) and 1,2, 3-trifluoro-5- (2-methoxyethoxy) -4-nitrobenzene (1.7g,6.8mmol) were reacted to form the title compound (1.02g, 32%); m/z =467[ M-1 ].

And C:5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3- (2-methoxyethoxy) benzene-1, 2-diamine：

Reduction of 2, 3-difluoro-N- (2-fluoro-4-iodophenyl) -5- (2-methoxyethoxy) -6-nitroaniline (1.017g,2.17mmol) using the conditions described in example 1 (step B) to form the title compound; m/z =337[ M-1 ].

Step D:l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- (2-methoxyethoxy) benzene Yl) cyclopropane-1-sulfonamides：

Following general procedure B, l-allyl-cyclopropanesulfonyl chloride (450mg,2.5mmol) was reacted with 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) -3- (2-methoxyethoxy) benzene-1, 2-diamine (219mg,2.5mmol) to give the title product (230mg, 78%); m/z =581[ M-1 ].

Step E:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- (2-methoxyethoxy) phenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide：

l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- (2-methoxyethoxy) phenyl) cyclopropane-l-sulfonamide (230mg,0.395mmol) and 4-methylmorpholine N-oxide (46mg,0.395mmol) were dissolved in THF (2 mL). Osmium tetroxide (0.039mmol,0.25mL,4% in H) was added at room temperature₂O) and the reaction mixture was stirred at room temperature for 16 hours. EtOAc was added and the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product.¹H NMR(300MHz,CDCl3)：7.36(dd,J=1.8&10.5Hz,1H),7.27(m,2H),6.56(dd,J=6.9&11.4Hz,1H),6.40(dt,J=5.7,7.5&12.9Hz,1H),4.17(m,2H),4.01(m,1H),3.78(m,2H),3.60(dd,J=3.6&11.1Hz,1H),3.47(m,1H),3.45(s,3H),2.36(dd,J=9.6&15.9Hz,1H),1.78(dd,J=2.4&15.6Hz,1H),1.45-1.25(m,2H),0.89(m,2H)。

Example 65

2, 4-dichloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) benzenesulfonamide:

synthesized by method a using the appropriate sulfonyl chloride, M/z =571[ M-1 ].

Example 66

2-chloro-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -4- (trifluoromethyl) benzenesulfonamide:

synthesized by method a using the appropriate sulfonyl chloride, M/z =605[ M-1 ].

Example 67

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2- (trifluoromethoxy) benzenesulfonamide:

synthesized by method a using the appropriate sulfonyl chloride, M/z =587[ M-1 ].

Example 68

4- (N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) sulfamoyl) benzoic acid:

Synthesized by method a using the appropriate sulfonyl chloride, M/z =584[ M-1 ].

Example 69

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) benzenesulfonamide:

synthesized by method a using the appropriate sulfonyl chloride, M/z =503[ M-1 ].

Example 70

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2-fluorobenzenesulfonamide:

synthesized by method a using the appropriate sulfonyl chloride, M/z =521[ M-1 ].

General procedure D: substitution of iodine atom:

in a microwave reactor, 1eq. aryl iodide, 1.5eq. boric acid or borate, 0.25eq. pdcl, will be included in the deoxygenated mixture of dioxane and water (3:1)₂(dppf) xDCM and 10eq. Anhydrous K₂CO₃The suspension of the powder was heated at 115 ℃ for 60 min. By aq₄Cl/THF extraction and Na₂SO₄The organic portion is dried. Flash column chromatography (Si, EtOAc/hexanes or CHCl) was used₃MeOH) purified the crude reaction product. Yield: 20 to 40 percent.

Example 71

N- (3, 4-difluoro-2- (2-fluoro-4-methylphenylamino) phenyl) cyclopropanesulfonamide;

general procedure D:¹H-NMR(500MHz,CDCl₃)：=7.38-7.36(m,1H),7.06-7.03(q,1H),6.92-6.90(1H),6.73-6.72(d,1H),6.63(s,1H,br),6.37-6.33(t,1H),5.54(s,1H,br),2.42-2.39(m,1H),2.25(s,3H),1.14-1.11(m,2H),0.94-0.90(m,2H)；m/z=355[M-1]。

where a racemic mixture of a chiral compound has been resolved into individual enantiomers, the phrase "substantially free" of epimers as used herein means an enantiomeric excess of at least 90%.

Example 72

N- (3, 4-difluoro-2- (2-fluoro-4- (1H-pyrazol-4-yl) phenylamino) phenyl) cyclopropanesulfonamide

Step A:2, 3-difluoro-N- (2-fluoro-4-iodophenyl) -6-nitroaniline:

to a solution of 2-fluoro-4-iodoaniline (11.40g,47mmol) in 100ml dry THF was added dropwise a 1M solution of LHMDS in THF (47mmol) (47ml) at 0 ℃. The color of the solution turned dark purple. The solution was transferred via cannula to a dropping funnel and this solution (containing the amine free base) was added portionwise to a solution of 2,3, 4-trifluoronitrobenzene (8.321g,47.0mmol) in anhydrous THF (50ml) at 0 ℃. After the addition was complete, the mixture was stirred at room temperature under argon for 15 hours. The volume of the solvent was reduced and then extracted with ethyl acetate and brine. The organic layer was dried over sodium sulfate, the solvent removed, and purified by flash chromatography (EtOAc/hexane 1:5, R)_f=0.58) the resulting dark oil was purified to give crude product which was dried in vacuo to brown solid (yield: 6.23g, 33.6%). M/z =393[ M-1 ]]^-。

And B:5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine

To a solution of nitro-diarylamine (6.23g,15.8mmol) in 300ml ethanol were added iron powder (13.74g,246mmol) and ammonium chloride (13.59g,254mmol) and the mixture was heated at 100 ℃ oil bath temperature with stirring for 14 hours. Filter and wash the residue twice with ethanol. The ethanol was removed in vacuo and the residue was extracted with ethyl acetate/1M NaOH solution. During the extraction, more precipitate formed was filtered and discarded. The combined organic layers were washed with brine and dried over sodium sulfate. Removing the solvent and separating from CHCl ₃The crude product was recrystallized from hexane (1: 50). A brown needle-like product (2.094g,66%,) was obtained. R_f=0.44(EtOAc/Hex1:3)。¹H-NMR(500MHz,CDCl₃)：=7.40-7.38(dd,1H,J=11.3Hz,J=1.5Hz).7.25-7.23(d,1H,J=8.5Hz),6.97-6.92(q,1H,J=9Hz),6.51-6.48(m,1H),6.24-6.21(t,1H,J=9Hz),5.3(s,1H,NH,br),3.80(s,2H,NH₂,br)；LRMS(ESI)：m/z=365[M+H]⁺。

And C:n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) cyclopropanesulphonamides：

Following general procedure a, 5, 6-difluoro-N1- (2-fluoro-4-iodophenyl) benzene-1, 2-diamine was reacted with cyclopropanesulfonyl chloride to give the desired product. (500MHz, CDCl)₃)：=7.38-7.37(d,1H),7.35-7.34(m,1H),7.27-7.26(m,1H),7.20-7.0(q,1H),6.68(s,1H,br),6.15-6.12(q,1H),5.65(s,1H,br),3.25-3.20(m,1H),2.4-2.3(m,2H),2.0-1.8(m,2H)；m/z=467[M-1]^-。

Step D:n- (3, 4-difluoro-2- (2-fluoro-4- (1H-pyrazol-4-yl) phenylamino) phenyl) cyclopropanesulfonamide:

general procedure C:¹H-NMR(500MHz,CDCl₃)：=8.00-7.90(m,2H),7.30-7.20(m,2H),7.15-7.10(m,1H),7.05-7.00(m,1H),6.70-6.60(m,1H),2.40-2.35(m,1H),1.05-1.0(m,2H),0.95-0.85(m,2H)；m/z=407[M-1]^-。

example 73

N- (3, 4-difluoro-2- (2-fluoro-4- (1-methyl-1H-pyrazol-4-yl) phenylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=7.95(s,1H),7.75(s,1H),7.30-7.20(m,2H),7.15-7.10(m,1H),7.05-7.00(m,1H),6.70-6.60(m,1H),3.95(s,3H),2.40-2.35(m,1H),1.05-1.0(m,2H),0.95-0.85(m,2H)；m/z=421[M-1]^-。

example 74

N- (3, 4-difluoro-2- (2-fluoro-4- (1H-pyrazol-3-yl) phenylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=7.90(s,1H),7.80(s,1H),7.30-7.20(m,2H),7.15-7.10(m,1H),7.05-7.00(m,1H),6.70-6.60(m,1H),3.95(s,3H),2.40-2.35(m,1H),1.05-1.0(m,2H),0.95-0.85(m,2H)；m/z=407[M-1]^-。

example 75

N- (3, 4-difluoro-2- (2-fluoro-4- (pyridin-4-yl) phenylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=8.62-8.61(d,2H),7.43-7.41(m,4H),7.23-7.22(m,1H),7.16-7.11(q,1H),6.61-6.58(t,1H),6.11(s,1H,br),2.53-2.50(m,1H),1.21-1.10(m,2H),1.02-0.99(m,2H)；m/z=418[M-1]^-。

example 76

N- (3, 4-difluoro-2- (2-fluoro-4- (pyridin-3-yl) phenylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,[D6]-DMSO)：=9.45(s,1H),8.91(s,1H),8.54(s,1H),8.07-8.06(d,1H),7.76-7.70(m,2H),7.46-7.34(m,2H),7.34-7.33(d,2H),6.80-6.78(m,1H),0.86-0.79(m,4H)；m/z=418[M-1]^-。

example 77

N- (2- (4-cyano-2-fluorophenylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide

In a microwave reactor, an aryl iodide (75.5mg,0.161mmol), CuCN (46.6mg,0.520mmol) and Pd (OAc) in 1ml of anhydrous DMF ₂(0.47mg) of the suspension was heated to 130 ℃ for 60 min. The mixture was extracted with brine/THF, and Na was used₂SO₄The organic portion is dried. Followed by flash column chromatography to yield a dark red semi-solid product (R)_f=0.42 (EtOAc/hexane 1: 1)). Yield: 15 percent. M/z =366[ M-1 ]]^-。

Example 78

N- (3, 4-difluoro-2- (3-fluorobiphenyl-4-ylamino) phenyl) cyclopropanesulfonamides

General procedure C:¹H-NMR(500MHz,CDCl₃)：=7.55-7.53(m,2H),7.45-7.3(m,5H),7.20-7.15(d,1H),7.13-7.10(q,1H),6.70(s,1H,br),6.60-6.55(t,1H),5.75(s,1H,br),2.53-2.50(m,1H),1.21-1.10(m,2H),1.02-0.99(m,2H)；m/z=417[M-1]^-。

example 79

N- (2- (3' -acetyl-3-fluorobiphenyl-4-ylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=8.6(s,1H),7.86-7.85(d,1H),7.68-7.66(d,1H),7.49-7.46(t,1H),7.38-7.33(m,2H),7.20-7.18(d,1H),7.09-7.03(q,1H),6.90(s,1H,br),6.57-6.54(t,1H),5.90(s,1H),br),2.61(s,3H),2.46-2.43(m,1H),1.15-1.13(m,2H),0.94-0.91(m,2H)；m/z=459[M-1]^-。

example 80

N- (2- (4' -cyano-3-fluorobiphenyl-4-ylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=7.68-7.66(m,2H),7.58-7.57(m,2H),7.38-7.35(m,2H),7.20-7.18(d,1H),7.18-7.02(q,1H),6.67(s,1H,br),6.58-6.54(t,1H),5.99(s,1H,br),2.47-2.44(m,1H),1.15-1.13(m,2H),0.94-0.91(m,2H)；m/z=442[M-1]^-。

example 81

N- (2- (3, 4' -difluorobiphenyl-4-ylamino) -3, 4-difluorophenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=7.44-7.37(m,3H),7.29-7.27(d,1H),7.11-7.05(m,4H),6.70(s,1H,br),6.53-6.50(t,1H),5.81(s,1H,br),2.47-2.44(m,1H),1.15-1.13(m,2H),0.94-0.91(m,2H)；m/z=435[M-1]^-。

example 82

N- (3, 4-difluoro-2- (3-fluoro-4' - (methylsulfonylamino) biphenyl-4-ylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,[D6]-DMSO)：=9.39(s,1H,br),7.63-7.60(m,3H),7.53-7.50(d,1H),7.30-7.23(m,4H),7.74-7.65(m,1H),2.99(s,3H),0.80-0.73(m,4H)；m/z=510[M-1]^-。

example 83

N- (3, 4-difluoro-2- (2-fluoro-4-methylphenylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,CDCl₃)：=7.38-7.36(m,1H),7.06-7.03(q,1H),6.92-6.90(1H),6.73-6.72(d,1H),6.63(s,1H,br),6.37-6.33(t,1H),5.54(s,1H,br),2.42-2.39(m,1H),2.25(s,3H),1.14-1.11(m,2H),0.94-0.90(m,2H)；m/z=355[M-1]^-。

example 84

4 '- (6- (cyclopropanesulfonamido) -2, 3-difluorophenylamino) -3' -fluorobiphenyl-3-carboxylic acid

General procedure C:¹H-NMR(500MHz,[D4]-MeOH)：=8.21(s,1H),7.93-7.91(d,1H),7.73-7.72(d,1H),7.47-7.43(m,2H),7.33-7.31(d,2H),7.15-7.12(q,1H),6.71-6.68(m,1H),2.51-2.46(m,1H),0.94-0.93(m,2H),0.88-0.87(m,2H)；m/z=499[M-1]^-。

example 85

N- (3, 4-difluoro-2- (3-fluoro-3' - (methylsulfonylamino) biphenyl-4-ylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,[D4]-MeOH)：=7.92(s,1H),7.46-7.34(m,5H),7.34-7.31(d,1H),7.29-7.22(m,1H),7.16-7.15(q,1H),6.74-6.71(m,1H),2.80(s,3H),2.54-2.51(m,1H),0.94-0.92(m,2H),0.91-0.90(m,2H)；m/z=510[M-1]^-。

example 86

N- (3, 4-difluoro-2- (3-fluoro-2' - (methylsulfonylamino) biphenyl-4-ylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,[D4]-MeOH)：=7.50-7.49(d,1H),7.40-7.32(m,4H),7.29-7.28(d,1H),7.26-7.10(m,2H),6.73-6.71(m,1H),2.80(s,3H),2.51-2.49(m,1H),0.94-0.92(m,2H),0.91-0.90(m,2H)；m/z=510[M-1]^-。

example 87

N- (3, 4-difluoro-2- (3-fluoro-4' - (trifluoromethoxy) biphenyl-4-ylamino) phenyl) cyclopropanesulfonamide

General procedure C:¹H-NMR(500MHz,[D4]-MeOH)：=7.69-7.67(d,2H),7.46-7.43(d,1H),7.36-7.33(m,4H),7.30-7.29(q,1H),6.73-6.72(m,1H),2.51-2.49(m,1H),0.94-0.92(m,2H),0.91-0.90(m,2H)；m/z=501[M-1]^-。

example 88

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2- (methylamino) ethanesulfonamide

General procedure D:¹HNMR(300MHz,CDCl₃)：9.01(br s,D₂o exchangeable, 1H),7.36(dd, J = 2.1)&10.5Hz,1H),7.27(m,1H),7.17(m,1H),7.03(dd,J=9.0&16.8Hz,1H),6.48(s,D₂O-exchangeable, 1H),6.31(dt, J =3.0, 8.7)&17.4Hz,1H),3.45(br t.2H).3.31(br s,2H),2.65(s.3H).1.80(br s,D₂O may be exchanged, 1H).

Example 89

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2- (2- (dimethylamino) ethylamino) ethanesulfonamide

General procedure D.¹H NMR(300MHz,CDCl₃)：7.35(m,1H),7.25(m,1H),7.18(d,J=8.7Hz,1H),7.02(dd,J=8.7&18.0Hz,1H),6.38(m,1H),6.18(dd,J=8.7&17.1Hz,1H),3.62(t,J=5.7&6.3Hz,2H),3.35(m,2H),3.26(m,2H),3.26(t,J=5.7&6.6Hz,2H),3.11(t,J=5.1&6.0Hz,2H),2.85(s,6H).

Example 90

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2- (ethyl (methyl) amino) ethanesulfonamide

General procedure D.¹H NMR(300MHz,(CDCl₃+D₂O))：7.39(dd,J=1.5&10.5Hz,1H),7.31(m,2H),7.07(dd,J=9.0&17.4Hz,1H),6.30(dt,J=2.4,9.0&17.4Hz,1H),3.55(t,J=6.9&7.8Hz,2H),3.38(br t,J=6.0&8.7Hz,2H),3.05(q,2H),2.69(s,3H),1.31(t,J=7.2Hz,3H)。

Example 91

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2- (4-methylpiperazin-1-yl) ethanesulfonamide

General procedure D.¹H NMR(300MHz,CD₃OD)：7.45(dd,J=2.1&10.8Hz,1H),7.30(m,2H),7.16(dd,J=9.6&17.7Hz,1H),6.39(dt,J=3.3,9.3&17.7Hz,1H),3.26(m,J=7.5Hz,2H),3.10(br m,6H),2.87(s,3H),2.82(t,J=7.5Hz,2H),2.48(br m,4H)。

In vitro biological Activity

Example 92

Generation of IC50 data

Preparing materials and reagents: human GST-MEK1 and constitutively active allele GST-MEK1^CA(with mutations Ser218Asp and Ser222Asp) was subcloned from the wild-type human MEK1 cDNA into the yeast expression vector pGEM4Z (Promega, Madison, Wis.). Expression of GST-MEK1 in E.coli ^CAAnd partially purified using glutathione sepharose 4B affinity resin (Amersham Pharmacia Biotech, Piscataway, NJ). The ERK2 allele was subcloned from MAPK2/ERK2 cDNA (wild type) in psusseamp (Upstate Biotechnology, inc., Waltham, MA) into vector pET21a (Novagen, Madison, WI) to yield an N-terminal histidine-tagged mouse ERK2 allele. Expression of ERK2 and purification to homogeneity [ Zhang,1993#33]. Myelin Basic Protein (MBP) was purchased from Gibco BRL (Rockville, Md.). EasyTides adenosine 5' -triphosphate (ATP) ([ gamma-³³P]) (NEN Perkin Elmer, Wellesley, Mass.) is a source of radiolabeling for all kinase reactions. Activated Raf-1 (truncated) and activated MAP kinase 2/ERK2 were purchased from Upstate, Inc. (Lake Placid, NY). 4-20% Criterion Precast gel was purchased from Bio-Rad (Hercules, Calif.).

And (3) measuring enzyme activity: compounds were diluted from dimethyl sulfoxide (DMSO) stock solutions to 1 XHNDE (20mM HEPES pH 7.2, 1mM MgCl)₂100mM NaCl, 1.25mM DTT, 0.2mM EDTA). A typical 25 microliter assay contains 0.002 nanomolar MEK1^CA0.02 nanomole ERK2, 0.25 nanomole MBP, 0.25 nanomolar unlabeled ATP and 0.1. mu. Ci [ gamma. ]³³P]ATP. This screening assay essentially comprised 4 additions. 5 μ l of the diluted compound was dispensed to a 96-well assay plate. Then, 10. mu.l of 2.5 Xenzyme cocktail (MEK 1 only) was added to each well ^CAAnd ERK2) followed by a 30 minute pre-incubation at ambient temperature. Then 10 μ l of 2.5x substrate cocktail (labeled and unlabeled ATP plus MBP) was added followed by incubation at ambient temperature for 60 minutes. Finally, 100 μ l 10% trichloroacetic acid (TCA) was added and incubated at room temperature for 30 min to stop the reaction and precipitate the radiolabeled protein product. The reaction product was collected onto a glass fiber 96-well filter plate pre-wetted with water and 1% pyrophosphate. The filter plate was then washed 5 times with water. The water was replaced with absolute ethanol and the plate was allowed to air dry at room temperature for 30 minutes. The back was sealed manually and 40 μ l of scintillation cocktail (scintillation cocktail) was dispensed to each well. The top seal was performed and the plates were counted in a TopCount at a rate of 2 seconds per well.

For some experiments, a truncated form of MEK requiring activation by Raf kinase was used.

Example 93

Generation of EC50 data

The effect of the compounds in the cells was detected by western blotting for phosphorylated ERK. MDA-MB-231 breast cancer cells were plated at 20,000 cells/well in 48-well plates and humidified CO at 37 °₂Culturing in an incubator. The next day, growth medium (DMEM +10% fetal bovine serum) was removed and replaced with starvation medium (DMEM +0.1% fetal bovine serum). Cells were incubated in starvation medium for 16 hours and then treated with a range of concentrations of compound for 30 minutes. After incubation with the compounds, cells were stimulated with 100ng/ml EGF for 5 minutes. The cells were then lysed and analyzed by western blotting using anti-phosphorylated ERK monoclonal antibodies. The signal was enhanced using a second antibody conjugated to a near infrared dye and detected on a LicorOdyssey scanner. Measuring the amount of signal intensity, this data being Used to generate dose response curves and EC50 calculations.

Legend: a, EC₅₀=<2.0nM；B,EC₅₀=2.0-15nM；C,EC₅₀=15nM-100nM；D,EC₅₀>100nM,IC₅₀<20μM；F,EC₅₀>100nM,IC₅₀>20μM

Legend: a, EC₅₀=<2.0nM；B,EC₅₀=2.0-15nM；C,EC₅₀=15nM-100nM；D,EC₅₀=100nM–200nM；E,EC₅₀>200 nM; ND = not determined

In vivo biological activity

Example 94

The compounds and compositions described herein are useful for treating or preventing one or more diseases, including, but not limited to, cancer, Inflammatory Bowel Disease (IBD), psoriasis, and Rheumatoid Arthritis (RA). The compounds and compositions described herein are also useful for once-daily or twice-daily oral treatment or prevention of one or more diseases including, but not limited to, cancer, IBD, psoriasis and RA.

In this example, in vivo testing of a compound of the following structure (compound a prepared as described herein) is described:

human tumors were implanted in nu/nu mice. Once the tumor size is about 100mm³I.e. compound a was administered orally for 14 days. Tumor Growth Inhibition (TGI) was determined as a reduction in tumor size in the treated groups compared to vehicle controls 14 days after treatment. The Time To Endpoint (TTE) was calculated as the time at which the tumor reached a particular endpoint volume or the time of any previous arrival in the final day of the study. The results of treatment were determined from% tumor growth delay (% TGD), which is defined as the% increase in the median TTE of mice receiving treatment compared to vehicle-treated control mice. Animals were also monitored for regression response. pERK levels in tumors and brain were determined by western blot and were correlated with plasma levels of compound a for pharmacodynamic/pharmacokinetic studies. Several tumor models were evaluated at different doses and dosing schedules. Treatment with 25 or 50mg/kg once daily (QD) in a375 melanoma tumor, Colo205 colon carcinoma and a431 epidermoid tumors showed statistically significant% TGD. Statistically significant TGI was observed for these tumor models as well as in HT29 colon cancer tumors, administered orally at 25mg/kg QD. The effect of different dosing regimens was evaluated in a375 xenografts. Although oral administration of 100mg/kg compound a once every two days showed statistically significant% TGD (91%), it was not therapeutically effective with QDs of 25mg/kg (143% TGD) or 50mg/kg (233% TGD). According to the% TGI determined, the twice daily (BID) dosing was also more effective than the QD dosing. Administration at 12.5mg/kg BID resulted in 79.5% TGI, compared to administration of compound a at 25mg/kg QD resulted in 51.7% TGI. Dosing at 25mg/kg BID resulted in 110.1% TGI, compared to 69.9% TGI given at 50mg/kg QD. Pharmacodynamic/pharmacokinetic studies in Colo205 xenografts Indicating that pERK formation is inhibited in tumors and minimal inhibition is observed in brain, indicating potent antitumor activity with limited CNS penetration.

Compound a is a potent MEK1/2 inhibitor that inhibits tumor cell growth in vitro and in vivo. BRAF status determines the sensitivity in anchorage-dependent growth rather than anchorage-independent growth, or in xenografts to compound-induced growth inhibition. Maintaining adequate MEK inhibition throughout the dosing interval appears to be more important than maintaining peak levels, since more frequent dosing is more effective. Depending on the xenograft results, compound a had a favorable pk profile in humans using the designed therapeutic dose of 20-40 mg/day in humans.

Example 94A

Inhibition of cancer cell Growth (GI)₅₀)

Anchorage-dependent growth inhibition was measured using CellTiterGlo reagent 48 hr after treating the cells cultured in 384-well plates with compound a. The anchorage-independent growth assay used MTS (methyl thiosulfonate) reagent 7 days after treatment of cells cultured in media containing 0.15% agarose or on non-adherent plates (a 431). Shown in the following table are growth inhibition values (GI)₅₀)。

Tumor cell lines	BRAF status	Anchorage dependent GI₅₀(nM±sd)	Anchorage-independent GI₅₀(nM±sd)
				A375 melanoma	V600E	67±12	68±34
Colo205 colon	V600E	74±45	33±16
				HT29 colon	V600E	70±12	Not determined
A431 epiderm sample	Is normal	>10,000	65±19

Example 94B

Anti-tumor xenograft activity

Implanting cells of A375 melanoma, Colo205 colon tumor, A431 epidermoid tumor or HT-29 colon tumor into female nu/nu mice to allow these tumors to grow to 100-³. Compound A or vehicle (25mg/kg, 50mg/kg or 100mg/kg) was administered orally once daily for 14 days. The mean tumor volumes of the vehicle and treatment groups were plotted and are shown in figure 1.

Example 94C

Tumor Growth Inhibition (TGI)25mg/kg QD

For the indicated tumor xenografts, the tumor growth inhibition of the group treated with 25mg/kg compound a was calculated. At the end of 14 days of once daily dosing, tumor growth inhibition was measured and calculated according to the following formula:

the range for a375 and Colo205 represents values from 2 independent studies.

Tumor xenografts	%TGI	P value
			A375 melanoma	52-72^**	<0.001
Colo205 colon	70-123^**	<0.001
			HT29 colon	56	<0.001
A431 epiderm sample	67	<0.001

^**Degradation was recorded during the test

Example 94D

ED in Colo205 xenografts₅₀

Colo205 tumor cells were implanted in male nu/nu mice. 10 days later, tumor size (range 126- ³) Animals were randomized and treated with paclitaxel (IV, QODx5), vehicle or compound a (PO, QDx 14).

Pharmacokinetic parameters were obtained from extrapolated values of Balb/c mice and lower dose groups dosed with 25mg/kg of Compound A and are given in the following table.

^*P<0.001

Example 94E

Tumor growth inhibition with A375 xenografts

Compound A50mg/kg QD, 25mg/kg BID, 50mg/kg QD and 12.5mg/kg BID were administered to A375 xenografted mice. The% TGI was calculated and plotted and is given in FIG. 2.

Example 94F

Plasma concentration in mice

Implanting A375 tumor cells into female nu/nu mice to allow tumor growth to 100-³. Compound A or vehicle (50mg/kg QD, 25mg/kg BID, 50mg/kg QD and 12.5mg @) is administered orally once daily (QD) or twice daily (BID)kg BID). At the end of 14 days of once daily dosing, tumor growth inhibition was measured and calculated according to the following formula:

AUC(μg·hr/ml)	132.5	117.0	66.5	78.0
					C_max(μg/ml)	23.8	10.2	11.9	7.8
C_min(μg/ml)	0.06	1.24	0.03	0.49
					C_minfree fraction (ng/ml)	0.117	2.48	0.059	0.986

Statistical significance = time series test

Example 94G

Inhibition of mouse xenograft tumor and brain MEK activity

Female nu/nu mice implanted with Colo205 tumor cells were administered a single dose of either vehicle or 2.5, 5, 10 or 25mg/kg of Compound A. The level of compound was determined in plasma samples, and pERK levels were determined in tumor and brain samples taken 2, 6, 12 and 24hr post-dose. pERK levels from western blots were quantified using LI-COR Odyssey, normalized to total ERK levels, and compared to vector treated levels to determine% MEK inhibition. MEK inhibition in each mouse tumor or brain and plasma concentrations of the corresponding compound a in the animals were plotted. Non-linear regression yields EC for MEK inhibition in tumors ₅₀Was 73 nM. Brain EC₅₀>5000nM。

Shown in FIG. 3 is a graph of plasma concentration (log nM) versus pERK% inhibition.

Preparation of capsules

Example 95A

Preparing a blue size 1 hard gelatin capsule comprising a dry powder blend composition wherein the structure isThe concentration of compound a (see the table shown in example 93 above) was 1mg and 10 mg.

Compound a was prepared as described herein and then micronized using a fluid energy Mill (helical Jet Mill, electrically grounded, milling chamber diameter 50 mm; 50 °.4 × 0.8mm nozzle ring; injector nozzle diameter 0.8mm, injector nozzle distance 3 mm). Compound a and a portion of microcrystalline cellulose were mixed, sieved through a #20 mesh sieve, and added to a diffusion-drum blender (V-blender). The remaining microcrystalline cellulose was sieved through a #20 mesh sieve, added to the material in the blender, and mixed. The croscarmellose sodium and sodium lauryl sulfate were sieved through a #20 mesh screen, added to the materials in the blender, and mixed. The powdered mixture was passed through a rotating impeller mill (Quadro coim) and added back to the mixer to continue mixing. The magnesium stearate was sieved through a #20 mesh screen and mixed with the powdered mixture. The powdered mixture was filled into size 1 capsules. For identification, 10mg capsules were banded.

The composition of the capsules is shown in the following table:

^atarget fill weights adjusted according to the actual efficacy of the mixture.

A typical batch formulation for 10,000 batches of 1mg capsules is as follows:

^atarget fill weights adjusted according to the actual efficacy of the mixture.

A typical batch formulation for 10,000 batches of 10mg capsules is as follows:

^atarget fill weights adjusted according to the actual efficacy of the mixture.

Example 95B

Preparing a blue size 1 hard gelatin capsule comprising a dry powder blend composition wherein the structure isThe concentration of compound B (see the table shown in example 93 above) was 1mg and 10 mg.

Compound B was prepared as described herein and micronized using a fluid energy Mill (helical Jet Mill, electrically grounded, milling chamber diameter 50 mm; 50 °.4 × 0.8mm nozzle ring; injector nozzle diameter 0.8mm, injector nozzle distance 3 mm). Compound B and a portion of microcrystalline cellulose were mixed, sieved through a #20 mesh sieve, and added to a diffusion drum mixer (V-blender). The remaining microcrystalline cellulose was sieved through a #20 mesh sieve, added to the material in the blender, and mixed. The croscarmellose sodium and sodium lauryl sulfate were sieved through a #20 mesh screen, added to the materials in the blender, and mixed. The powdered mixture was passed through a rotating impeller mill (Quadro coim) and added back to the mixer to continue mixing. The magnesium stearate was sieved through a #20 mesh screen and mixed with the milled powder blend. The powdered mixture was filled into size 1 capsules. For identification, 10mg capsules were bundled.

The following table shows the composition of the capsules:

in vivo activity in humans

Example 96

Administration of the capsules described in example 95A to human cancer patients

A single dose of either 1mg or 10mg of the capsule composition described above in example 95A was administered to a human cancer patient. For a 2mg dose, 2x1mg capsules were administered to the patient; for a 4mg dose, 4x1mg capsules were administered to the patient; for a 6mg dose, the patient was administered a 6x1mg capsule; for a 10mg dose, 1x10mg capsules were administered to the patient; for the 20mg dose, 2x10mg capsules were administered to the patient.

The concentration-time profile was monitored and is given in figure 4 and the following table:

crystalline polymorphs

Example 97: preparation of N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was prepared according to the procedure described above (see published International patent application WO 2007/014011) and as outlined below.

Step A: 2-fluoro-N- (2,3, 5-trifluoro-6-nitrophenyl) -4-iodoaniline

1.0M lithium hexamethyldisilazide (LiN (SiMe) was added under nitrogen at-78 deg.C₃)₂) The "LHMDS" solution (15.37mL,15.37mmol) was slowly added to a stirred solution of 2-fluoro-4-iodoaniline (3.64g,15.37mmol) in anhydrous THF (100mL) and stirring was continued for an additional 1 hour at-78 deg.C. 2,3,4, 6-Tetrafluoronitrobenzene was added and the reaction mixture was allowed to warm to room temperature and stirring was continued for an additional 16 hours. Ethyl acetate (200mL) was added and the organic phase was washed with water, dried over sodium sulfate and further purified by column chromatography to yield the product as a yellow solid (3.75g, 59.24%). M-H ⁺：410.9。¹H NMR(DMSO,300MHz)：6.85(t,1H)；7.38(d,1H)；7.62(m,2H)；8.78(s,1H)。

And B: 2-fluoro-N- (2, 3-difluoro-5-methoxy-6-nitrophenyl) -4-iodoaniline

A stirred solution of (2-fluoro-4-iodo-phenyl) - (2,3, 5-trifluoro-6-nitro-phenyl) -amine (1.23g,3mmol) in anhydrous THF (25ml) was cooled to-78 ℃ under nitrogen and 25% sodium methoxide solution (0.68ml,0.3mmol) was slowly added. The reaction mixture was allowed to warm to room temperature and stirring was continued for a further 16 hours. TLC indicated incomplete reaction. Ethyl acetate (100mL) was added to the reaction mixture, the organic layer was washed with water, dried over sodium sulfate, and further purified by column chromatography to give the desired compound (0.6g,47.6%) as a yellow solid. M/z =424[ M + H%]⁺。

And C: 5, 6-difluoro-N¹- (2-fluoro-4-iodophenyl) -3-methoxybenzene-1, 2-diamine

Ammonium chloride (1.18g,20.16mmol) and iron powder (1.15g,21.44mmol) were added to a stirred solution of (2, 3-difluoro-5-methoxy-6-nitro-phenyl) - (2-fluoro-4-iodo-phenyl) -amine (0.57g,1.34mmol) in ethanol (20 mL). The mixture was stirred at reflux for 16 h, cooled to room temperature, filtered through celite, and the filtrate was concentrated to dryness. The resulting residue was dissolved in ethyl acetate, washed with water, dried over sodium sulfate, and further purified by crystallization from ethanol to yield an off-white solid (0.47g, 90.3%). M-H ⁺：393.2。¹H NMR(DMSO,300MHz)：3.76(s,3H)；6.1(t,1H)；6.8-7.0(m,1H)；7.2(d,1H)；7.35(s,1H)；7.42(d,1H)。

Step D: 1-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-1-sulfonamide

To 5, 6-difluoro-N¹To a stirred solution of- (2-fluoro-4-iodophenyl) -3-methoxybenzene-1, 2-diamine (1eq) in anhydrous pyridine (5ml/mmole) was added 1-allyl-cyclopropanesulphonyl chloride (1-5 eq). The reaction mixture was stirred at 40 ℃ for 48 hours. The reaction mixture was partitioned with water and ethyl acetate. The organic layer was washed with brine and dried (MgSO)₄) And concentrated under reduced pressure.The residue was purified by flash column chromatography over silica gel to give the title product.¹H NMR(CDCl₃,300MHz)：7.417(dd,1H),7.309(s,1H),7.25(m,1H),6.89(m,1H),6.52(m,1H),6.427(m,1H),6.03(s,lH),5.668(m,1H),5.11(t,1H),3.9(s,3H),2.75(d,2H),1.21(m,2H),0.767(m,2H)。

Step E: n- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

l-allyl-N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-l-sulfonamide (97mg,0.18mmol) and 4-methylmorpholine N-oxide (21mg,0.18mmol) were dissolved in THF (8 mL). Osmium tetroxide (0.018mmol,0.13mL,4% in H) was added at room temperature₂O) and the reaction mixture was stirred at room temperature for 16 hours. Ethyl acetate was added and the organic phase was washed with water and dried (MgSO)₄) And concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: EtOAc/MeOH) to give the title product (0.80g, 78%). ¹H NMR(CDCl₃,300MHz)：7.38(dd,J=1.7&10.3Hz,1H),7.26(m,1H),7.14(s,1H),6.87(s,1H),6.53(dd,J=6.8&11.4Hz,1H),6.43(m,1H),4.06(m,1H),3.89(s,3H),3.63(dd,J=3.7&11.1Hz,1H),3.49(dd,J=6.4&11.1Hz,1H),2.3(dd,J=9.7&16.1Hz,1H),1.77(dd,J=1.9&16.0Hz,1H),1.37(m,1H),1.25(m,1H),1.21(m,2H),0.86(m,2H)；m/z=571[M-1]^-。

Example 98: preparation of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

The racemic mixture was separated by chiral HPLC to give the pure S isomer.¹H NMR(CDCl₃,300MHz)：7.38(dd,J=1.7&10.3Hz,1H),7.26(m,1H),7.14(s,1H),6.87(s,1H),6.53(dd,J=6.8&11.4Hz,1H),6.43(m,1H),4.06(m,1H),3.89(s,3H),3.63(dd,J=3.7&11.1Hz,1H),3.49(dd,J=6.4&11.1Hz,1H),2.3(dd,J=9.7&16.1Hz,1H),1.77(dd,J=1.9&16.0Hz,1H),1.37(m,1H),1.25(m,1H),1.21(m,2H),0.86(m,2H)；m/z=571[M-1]^-。

Example 99: preparation of N- (R) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

The racemic mixture was separated by chiral HPLC to give the pure R isomer.¹H NMR(CDCl₃,300MHz)：7.38(dd,J=1.7&10.3Hz,1H),7.26(m,1H),7.14(s,1H),6.87(s,1H),6.53(dd,J=6.8&11.4Hz,1H),6.43(m,1H),4.06(m,1H),3.89(s,3H),3.63(dd,J=3.7&11.1Hz,1H),3.49(dd,J=6.4&11.1Hz,1H),2.3(dd,J=9.7&16.1Hz,1H),1.77(dd,J=1.9&16.0Hz,1H),1.37(m,1H),1.25(m,1H),1.21(m,2H),0.86(m,2H)；m/z=571[M-1]^-。

Example 100: preparation of crystalline polymorph A of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

Preparation i) N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -L- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (216.10g) was added to a 4L Erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Ethyl acetate (about 600mL, from Fisher) was added. Heating was started and stirring was carried out to form a brown suspension. The mixture was slightly refluxed and additional ethyl acetate (about 200mL) was added to achieve complete dissolution to give a dark brown solution. Heptane (from Acros) was slowly added in portions to the refluxing solution at a rate such that all the precipitate formed on each addition dissolved rapidly and reflux was maintained. When 2L of heptane was added to this solution, the solid formed dissolved very slowly at reflux. The heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature over 16h with stirring. During aging, a thick layer of crystalline material forms around the glass surface. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a 25cm buchner funnel fitted with Whatman #1 filter media. The collected crystals were washed with heptane (1L) and allowed to air dry under vacuum. Further drying of this crystal at 40 ℃/<1 torr for 20h yielded a pink crystalline solid product (160.99g, 77.2%).

Preparation ii) N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (13.2g) and ethyl acetate (30mL) were added to an erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Stirring was started and heated to weak reflux to achieve complete dissolution to give a dark brown solution. Heptane was slowly added to the refluxing solution in portions at a rate that all the precipitate formed on each addition dissolved rapidly and maintained reflux until the addition of heptane to the solution caused the solids formed to dissolve very slowly at reflux (-90 mL heptane). The heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature over 16h with stirring. During aging, a thick layer of crystalline material forms around the glass surface. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a buchner funnel fitted with Whatman #1 filter media. The collected crystals were washed with heptane and allowed to air dry under vacuum. This crystal was further dried at 40 ℃/<1 torr for 20h to yield a pink crystalline solid product.

Preparation iii) N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (44.8g) and ethyl acetate (750mL) were added to an erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Stirring was started and heated to weak reflux to achieve complete dissolution to give a dark brown solution. Hexane was slowly added portion by portion to the refluxing solution at a rate that all the precipitate formed on each addition dissolved rapidly and maintained reflux until the addition of hexane to the solution caused the solids formed to dissolve very slowly at reflux (-2L hexane). The heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature over 16h with stirring. During aging, a thick layer of crystalline material forms around the glass surface. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a buchner funnel fitted with Whatman #1 filter media. The collected crystals were washed and allowed to air dry under vacuum. This crystal was further dried at 40 ℃/<1 torr for 20h to yield a pink crystalline solid product.

Example 101: preparation of crystalline polymorph of N- (R) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

N- (R) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -L- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (216.10g) was added to a 4L Erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Ethyl acetate (ca. 600mL) was added. Heating was started and stirring was carried out to form a brown suspension. The mixture was slightly refluxed and additional ethyl acetate (about 200mL) was added to achieve complete dissolution to give a dark brown solution. Heptane was slowly added portionwise to the refluxing solution at a rate that all the precipitate formed on each addition dissolved rapidly and maintained reflux. When 2L of heptane was added to this solution, the solid formed dissolved very slowly at reflux. The heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature over 16h with stirring. During aging, a thick layer of crystalline material forms around the glass surface. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a 25cm buchner funnel fitted with Whatman #1 filter media. The collected crystals were washed with heptane (1L) and allowed to air dry under vacuum. The crystals were further dried at 40 ℃/<1 torr for 20 h.

Example 102：IC50Generation of data

Preparing materials and reagents: human GST-MEK1 and constitutively activated allele GST-MEK1^CA(with mutations Ser218Asp and Ser222Asp) was subcloned from the wild-type human MEK1 cDNA into the yeast expression vector pGEM4Z (Promega, Madison, Wis.). Expression of GST-MEK1 in E.coli^CAAnd partially purified using glutathione sepharose 4B affinity resin (Amersham Pharmacia Biotech, Piscataway, NJ). The ERK2 allele was subcloned from MAPK2/ERK2 cDNA (wild type) in psusseamp (Upstate Biotechnology, inc., Waltham, MA) into vector pET21a (Novagen, Madison, WI) to yield an N-terminal histidine-tagged mouse ERK2 allele. Expression of ERK2 and purification to homogeneity [ Zhang,1993#33]. Myelin Basic Protein (MBP) was purchased from Gibco BRL (Rockville, Md.). EasyTides adenosine 5' -triphosphate (ATP) ([ gamma-³³P]) (NEN Perkin Elmer, Wellesley, Mass.) is a source of radiolabeling for all kinase reactions. Activated Raf-1 (truncated) and activated MAP kinase 2/ERK2 were purchased from Upstate, Inc. (Lake Placid, NY). 4-20% Criterion Precast gel was purchased from Bio-Rad (Hercules, Calif.).

And (3) measuring enzyme activity: compounds were diluted from dimethyl sulfoxide (DMSO) stock solutions to 1 XHNDE (20mM HEPES pH 7.2, 1mM MgCl) ₂100mM NaCl, 1.25mM DTT, 0.2mM EDTA). A typical 25 microliter assay contains 0.002 nanomolar MEK1^CA0.02 nanomolar ERK2, 0.25 nanomolar MBP, 0.25 nanomolar unlabeled ATP, and 0.1 μ Ci [ γ ] Ci³³P]ATP. This screening assay essentially comprised 4 additions. 5 μ l of the diluted compound was dispensed to a 96-well assay plate. Then, 10. mu.l of 2.5 Xenzyme cocktail (MEK 1 only) was added to each well^CAAnd ERK2) followed by a 30 minute pre-incubation at ambient temperature. Then 10 μ l of 2.5x substrate mix (labeled and unlabeled ATP plus MBP) was added followed by incubation at ambient temperature for 60 minutes. Finally, 100 μ l 10% trichloroacetic acid (TCA) was added and incubated at room temperature for 30 min to stop the reaction and precipitate the radiolabeled protein product. The reaction product was collected onto a glass fiber 96-well filter plate pre-wetted with water and 1% pyrophosphate. The filter plate was then washed 5 times with water. With anhydrous ethanolThe water was replaced and the plate was allowed to air dry at room temperature for 30 minutes. The back was sealed manually and 40 μ l of scintillation fluid was dispensed to each well. The top seal was performed and the plates were counted in a TopCount at a rate of 2 seconds per well. For some experiments, a truncated form of MEK requiring activation by Raf kinase was used.

Example 103: generation of EC50 data

The effect of the compounds in the cells was detected by western blotting for phosphorylated ERK. MDA-MB-231 breast cancer cells were plated at 20,000 cells/well in 48-well plates and humidified CO at 37 °₂Culturing in an incubator. The next day, growth medium (DMEM +10% fetal bovine serum) was removed and replaced with starvation medium (DMEM +0.1% fetal bovine serum). Cells were incubated in starvation medium for 16 hours and then treated with a range of concentrations of compound for 30 minutes. After incubation with the compounds, cells were stimulated with 100ng/ml EGF for 5 minutes. The cells were then lysed and analyzed by western blotting using anti-phosphorylated ERK monoclonal antibodies. The signal was enhanced using a second antibody conjugated to a near infrared dye and detected on a Licor Odyssey scanner. The amount of signal intensity was determined and this data was used to generate dose response curves and EC50 calculations.

Example 104: activity data of the Compounds

The compounds described in examples 1, 2 and 3 were tested in the above assay. The results are summarized in the following table (A, EC)₅₀=<2.0nM；B,EC₅₀=2.0-15nM)：

Example 105: XRPD data

XPRD was performed on an Inel XRG-3000 diffractometer equipped with a curved position-sensitive detector (curved position-sensitive detector) with a 120 ° 2 θ range. Real-time data was acquired using Cu ka radiation at a resolution of 0.03 ° 2 θ. The voltage and amperage of the tube were set to 40kV and 30mA, respectively. The graph is shown from 2.5 to 40 ° 2 θ to facilitate direct comparison of the graphs. A sample of (S) -N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -l- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (synthesized as described herein) was prepared for analysis by filling the sample into a thin-walled glass capillary tube. Each capillary was moved onto a motorized goniometer head, allowing the capillary to be rotated during data acquisition. The samples were analyzed for 5 minutes. Instrument calibration was performed daily using silicon reference standards. FIG. 5 is a powder X-ray diffraction (PXRD) pattern of form A N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. FIG. 7 is a powder X-ray diffraction (PXRD) pattern of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide form A (upper panel) and amorphous form (lower panel).

Example 106: differential Scanning Calorimetry (DSC)

The analysis was performed on a TA instruments differential scanning calorimeter Q1000. The instrument was calibrated using indium as a reference material. The samples were placed in a standard aluminum DSC pan with a non-crimped lid (non-crimped) construction and the weight was accurately recorded. To determine the glass transition temperature (T) of amorphous materials_g) The cell was cycled several times between-40 ℃ and 140 ℃. The final temperature was raised to 150 ℃. Inflection point report T from last cycle conversion_g. FIG. 6 is a modulated DSC thermogram of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form A).Normalized heat flow in watts per gram (W/g) is plotted against measured sample temperature in degrees celsius.

Example 107: dynamic steam adsorption/Desorption (DVS)

Moisture adsorption/desorption data were collected on a VTI SGA-100 steam adsorption analyzer. Adsorption/desorption data were collected at 10% RH intervals, under a nitrogen purge, over a range of 5% to 95% Relative Humidity (RH). The samples were not dried prior to analysis. The equilibrium criteria used for the analysis were a weight change of less than 0.0100% in 5 minutes, and if the weight criteria were not met, the maximum equilibration time was 3 hours. The initial moisture content of the sample was not corrected for data. Sodium chloride and polyvinyl pyrrolidine (polyvinypyrolidine) were used as calibration standards. FIG. 8 shows DVS isotherms for N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form A). During the experiment, this material showed negligible weight change.

Example 108: thermogravimetry (TG)

Analysis was performed on a TA instruments 2950 thermogravimetric analyzer. The calibration standards were nickel and Alumel^TM. Each sample was placed in an aluminum sample pan and inserted into a TG furnace. The sample was equilibrated at 25 ℃ and then heated at a heating rate of 10 ℃/min under a stream of nitrogen until the final temperature was 350 ℃. Figure 9 is a TG thermogram of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (form a) showing negligible weight loss up to 140 ℃, indicating that polymorph form a is unsolvated.

Example 109: in vitro cancer screening

In RPMI 164 containing 5% fetal bovine serum and 2mM L-glutamine0 medium to human tumor cell lines. Cells were seeded in 96-well microplates at 100 μ Ι _ at a plating density ranging from 5,000 to 40,000 cells/well depending on the doubling time of each cell line. After cell inoculation, 5% CO at 37 ℃₂These microplates were incubated for 24h, 95% air and 100% relative humidity, then N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was added.

To represent the measurements of the cell population for each cell line upon addition of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (Tz), after 24h, two plates of each cell line were fixed in situ with TCA. N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was dissolved in dimethyl sulfoxide at 400 times the desired final maximum test concentration and stored frozen prior to use. At the time of addition, an aliquot of the frozen concentrate was thawed and diluted to twice the desired final maximum test concentration with complete medium containing 50 μ g/ml gentamicin. To provide a total of 5 concentrations plus control, additional 4-fold, 10-fold, or 1/2 log-series dilutions were made. 100 μ l aliquots of these different dilutions were added to appropriate microtiter wells already containing 100 μ l of medium to give the desired final concentrations.

After addition of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, 5% CO at 37 deg.C₂These plates were incubated for an additional 48h, 95% air and 100% relative humidity. For adherent cells, the assay was terminated by the addition of cold TCA. Cells were fixed in situ by gently adding 50. mu.l of cold 50% (w/v) TCA (final concentration, 10% TCA) and incubated for 60 min at 4 ℃. The supernatant was discarded, and the plate was washed 5 times with tap water and air-dried. A0.4% (w/v) Sulforhodamine B (SRB) solution (100. mu.l) in 1% acetic acid was added to each well and the plates were incubated for 10 minutes at room temperature. After dyeing, unbound dye was removed by washing 5 times with 1% acetic acid and the plates were air dried. After that The bound dye was dissolved with 10mM tris (hydroxymethyl) aminomethane (trizma base) and the absorbance was read at 515nm wavelength using an automatic plate reader. For suspension cells, the procedure was the same except that the cells settled at the bottom of the well were fixed by gently adding 50 μ l 80% TCA (final concentration, 16% TCA) to terminate the assay. Test growth (Ti) using 7 absorbance measurements [ time zero (Tz), control growth (C) and the presence of 5 concentration levels of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide]The% growth at each drug concentration level was calculated. The% growth inhibition was calculated as follows:

three dose response parameters were calculated. 50% growth inhibition (GI50) was calculated from [ (Ti-Tz)/(C-Tz) ] x100=50, which is the concentration that causes a 50% decrease in net protein increase (as measured by SRB staining) in control cells during drug incubation. The N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide concentration that caused complete growth inhibition (TGI) was calculated from Ti = Tz. LC50 (drug concentration causing a 50% reduction in the measured protein at the end of drug treatment compared to the initial) representing the net reduction in treated cells was calculated from [ (Ti-Tz)/Tz ] x100= -50. If the activity level is reached, calculating each value of the three parameters; however, if this effect is not met or exceeded, the parameter is expressed as being above or below the highest or lowest concentration tested.

The indicated cell lines were examined and the following results are given, corresponding to panels of leukemia, non-small cell lung cancer, colon cancer, CN cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer.

Example 110: in vitro antiproliferative activity

In this example, the following effects of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide were examined: (1) inhibitory Activity on growth of several tumor cell lines with different mutations (GI)₅₀) (ii) a (2) Inhibitory Activity on growth of several B-Raf mutant cell lines (GI)₅₀) (ii) a (3) Effects on anchorage-independent cell growth; (4) effects on the cell cycle; and (5) toxic effects on primary hepatocytes and kidney cells.

Cell culture/growth inhibition assay

Human melanoma a375 cells and human colon cancer Colo205 cells were obtained from ATCC (Manassas, VA). A375 cells were cultured in DMEM supplemented with 10% fetal bovine serum, glutamine (2mM), penicillin (100U/ml) and streptomycin (100. mu.g/ml). At 37 deg.C, 5% CO₂And culturing the cells under 100% humidity. Colo205 cells were cultured in RPMI supplemented with 10% fetal bovine serum, glutamine (2mM), penicillin (100U/ml) and streptomycin (100. mu.g/ml). For growth inhibition assays, cells were plated in white 384-well microplates at 1000 cells/20 μ l/well. After 24hr, 5 μ l of 5X drug stock solution was added. All drugs were initially formulated as 200X stock solutions in DMSO, resulting in a final DMSO concentration of 0.5%. Cells were incubated at 37 ℃ for 48hr and ATP levels were determined using CellTiterGlo (Promega, Madison, Wis.). Adenylate Kinase (AK) release was measured using Toxilight (Cambrex, walker, MD). Nonlinear curve fitting was performed using GraphPad Prism 4(GraphPad software, San Diego, CA). 4-amino-8- ((2R,3R,4S,5R) -3, 4-dihydroxy Yl-5- (hydroxymethyl) tetrahydrofuran-2-yl) pyrido [2,3-d]Pyrimidin-5 (8H) -one (VRX-14686) is a cytotoxic agent used as a reference compound.

Growth inhibition(%) = (vehicle only control (RLU) -N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide RLU)/(vehicle only control RLU-1 μ M N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide RLU); according to the growth arrest caused by N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, ATP levels were measured.

Cell viability(%) = (N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide RLU-10 μ M VRX-14686 RLU)/(vehicle only control RLU-10 μ M tamoxifen RLU); based on cell killing by VRX-14686, where ATP levels are measured.

Cell killing(%) = (N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide RLU-vehicle only control RLU)/(10 μ M tamoxifen RLU-vehicle only control RLU); AK release was measured according to cell killing by tamoxifen.

RLU = relative luminescence unit

Evaluation of cell cycle arrest

A375 cells were plated at 10,000 cells/200. mu.l/well in 96-well microplates. After 24hr, the cells were approximately 50% confluent and 50 μ l of 5X drug stock solution was added. After an additional 24hr, the cells were trypsinized, fixed in 200. mu.l preferr (Anatech, Battle Creek, MI) and stored at 4 ℃ overnight. Cells were then washed in PBS, permeabilized, stained in 0.1% Triton X-100, 200. mu.g/ml DNase-free ribonuclease and 25. mu.g/ml propidium iodide (Molecular Probes, Sunnyvale, Calif.), and analyzed on Guava PCA-96(Guava Technologies, Foster City, Calif.). Data were analyzed using ModFit LT (version 3.0, Verity, Topsham, ME).

(1) Evaluation of growth inhibition of anchorage-independent cells

The wells of an "ultra-low adhesion" plate (Corning, Acton MA) were filled with 60. mu.l of 0.15% agarose solution in intact RPMI. Then, 60 μ l of complete RPMI containing 9000 Colo205 cells in 0.15% agarose was added to each well. After 24hr, 60 μ l of 3X drug solution in complete RPMI without agarose was added. After 7 days, 36. mu.l of 6X MTS reagent (CellTiter 96 Aqueous, Promega, Madison, Wis.) was added to each well. After 2hr at 37 deg.C, the absorbance was measured at 490nm on an M5 plate reader (Molecular Devices, Sunnyvale, Calif.). Nonlinear curve fitting was performed using GraphPad Prism 4.

(2) Growth inhibition of MEK-dependent cancer cell Growth (GI)₅₀)

Log phase dividing B-Raf mutant cells a375 (human melanoma), a431 (melanoma), Colo205 (colon carcinoma), HT29 (colorectal adenocarcinoma), MDA-MB231 (breast adenocarcinoma) and BxPC3 (pancreatic adenocarcinoma) were contacted with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide for 48hr and ATP content was determined. 100% growth arrest was determined using 1 μ M N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

The following table shows the average GI of each cell line from at least three experiments₅₀Values and showed that N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide caused growth inhibition in three B-Raf mutant cell lines (a375, Colo205, and HT29) and one ras/Raf/MEK/MAPK pathway wild-type cell line (a431) with an average potency of 79nM (+ -9 nM).

Cell lines	Mean value of	Standard deviation of	C.V.
				A375	71nM	12.1nM	17%
A431	86nM	25.4nM	30%
				Colo205	89nM	40.1nM	45%
HT29	70nM	12.2nM	18%
				MDA	>1uM
BxPC3	>1uM

In a separate study, B-Raf mutant cells a375 (human melanoma), SKMel28 (human melanoma), and Colo205 (human colon carcinoma) that are dividing in log phase were contacted with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide for 48 hrs and ATP content was determined. The following table shows the GI of each cell line ₅₀It was shown that N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide caused growth inhibition, which had EC inhibition similar to that of its MEK₅₀Efficacy of the value.

Cell lines	GI₅₀(nM)
		A375	56
SK Mel28	105
		Colo205	27

FIGS. 10A and 10B show growth arrest of dividing A375 cells in log phase in contact with increasing concentrations of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. The assay cells were assayed for ATP content. 100% growth arrest was determined using 1 μ M N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Cell supernatants were analyzed for cytotoxic lysis by measuring Adenylate Kinase (AK) release. Log phase dividing a375 cells were contacted with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and PD-325901 for 48 hr. (100% cell killing was determined using 20 μ M tamoxifen). The results are shown in fig. 11. This data indicates that N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide causes non-toxic growth arrest in several susceptible human cancer cell lines, as measured by i) growth arrest assay (ATP quantification); and ii) lack of cytotoxic cell lysis (AK release). For all cell lines tested, lack of AK release was demonstrated.

Anchorage-independent growth inhibition

Adherent independent growth of Colo205, a375, and MDA-MB231 cells in contact with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide for 7 days was quantitatively evaluated in a 96-well microplate format. Viability was determined by MTS assay. GI (GI tract)₅₀The values are as follows:

cell lines	Mean value of	Standard deviation of	C.V.
				Colo205	40nM	8.1nM	20%
A375	84nM	17.2nM	21%
				MDA-MB231	81nM	55.6nM	69%

FIGS. 12A-12C show the Growth Inhibition (GI) of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (A) on human colorectal carcinoma Colo205 cells₅₀=11 nM); (B) growth inhibition of A375 cells (GI)₅₀=22nM) and (C) inhibition of MDA-MB231 cells, which showed no growth arrest by N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide in a two-dimensional adherence dependent assay.

Log phase dividing a375 cells were contacted with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (1uM) for 48hr and cell supernatants were analyzed for growth inhibition (ATP content) and cytotoxic lysis (AK release). In the vehicle control wells only, 100% viability was determined (ATP content assay). The results shown in the table below indicate that N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide causes non-toxic growth arrest in B-Raf mutant human melanoma a375 cells.

	% control
		ATP, cell viability	27%
AK, cell killing	4%

Anchorage-independent growth inhibition

Anchorage-independent growth was quantitatively evaluated in 96-well microplate format. FIG. 13A shows growth inhibition, GI, of Colo205 cells in human colorectal cancer₅₀Values were not 6nM and 11nM, respectively. FIG. 13B shows growth inhibition of A375 cells, GI₅₀Values were 5nM and 22 nM.

N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropane YF) cell cycle analysis of growth arrest caused by cyclopropane-1-sulfonamide

MEK inhibition in A375 cells has been shown to cause cell cycle arrest in the G1/S phase.

Log phase dividing a375 cells were contacted with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide for 24hr and the percentage of cells stained for stage-dependent intracellular DNA amounts was determined using flow cytometry.

The following table shows the percentage distribution of cells in each growth phase among the cells treated with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and the control (vehicle only).

FIGS. 14A and 14B show the effect of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on cell cycle progression, indicating that contacting A375 cells with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide causes cell cycle arrest in the G1 phase, as indicated by depletion of cells in both the G2 and S phases.

Evaluation of Primary hepatocyte and Kidney cytotoxicity

Cryopreserved rat hepatocytes were obtained from CellzDirect (Austin, TX) and plated in collagen-coated 96-well plates according to the manufacturer's instructions. 4hr after plating, drug (final DMSO concentration 0.5%) was added.

Plated human hepatocytes were obtained from CellzDirect and processed according to the manufacturer's instructions.

Cryopreserved human Renal Proximal Tubule Epithelial Cells (RPTEC) were obtained from Cambrex and processed according to the manufacturer's instructions. Cells were allowed to proliferate for 4 days and then plated at 50,000 cells/well in 96-well plates for drug exposure.

After 48hr, supernatant AK levels were determined using Toxilight and cellular ATP levels were determined using CellTiterGlo. Complete kill values were determined using 15. mu.M VRX-14686.

The results are shown below. Very little cell lysis was observed. In freshly plated primary human hepatocytes, the lowest toxicity (81% survival) was observed at 30 μ M N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. RPTEC cells showed dose-dependent ATP depletion and significant cell lysis at 30 μ M.

The above data indicate that (1) N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide inhibits cell growth and division in selected human cancer cells, the GI thereof in an anchorage-dependent proliferation assay₅₀Values in the 70-89nM range, as determined in the cell lysis assay without toxicity; (2) n- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide inhibits cell growth and division in selected human cancer cells, GI in an anchorage-dependent and independent proliferation assay₅₀Values were 51nM and 22nM, respectively; (3) n- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide causes G1 arrest and inhibits anchorage-independent growth in a375 cells, providing evidence of anticancer activity in a physiologically relevant in vitro model; and (4) N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide showed little cytotoxicity against primary normal human hepatocytes, human renal proximal tubule epithelial cells, and rat hepatocytes.

Example 111: pharmacokinetics of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide in cancer patients after multiple dosing

R_ac: cumulative index

^*Due to limited sampling time, inaccurate evaluation

After multiple administrations of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide at 2, 4 or 6 mg/subject, N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was readily absorbed, with an average T of_maxThe range is 1.33 to 1.50 hr. Average C_max、C_τAnd AUC values increase with dose in a manner proportional to dose. For C_maxThe cumulative index ranged from 1.49 to 1.76, for AUC, the cumulative index ranged from 1.90 to 2.07, indicating a modest degree of accumulation. Although the half-life cannot be accurately determined due to limited sampling time after multiple administration, the half-life is expected to be longer than 22hr after multiple administration, according to the cumulative index. These half-life values are significantly longer than those observed in mouse efficacy models (the typical range observed is 2 to 3 hr). Furthermore, at all doses, encouraging peak-to-valley ratios were observed.

Example 112: pharmacokinetics of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide in healthy volunteers after multiple dosing

R_a: cumulative index

^*Inaccurate evaluation due to limited sampling time

After multiple administrations of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide at 10 or 20 mg/subject, N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was readily absorbed, with its average T_maxThe range is 2.00 to 2.25 hr. Average C_max、C_τAnd AUC values increase with dose. For C_maxTired ofThe product index ranged from 1.14 to 1.23, for AUC, the cumulative index ranged from 1.24 to 1.29, indicating that the accumulation was not significant. The half-lives of both dosing regimens are similar, ranging from 13 to 15 hr. These half-life values are shorter than those observed in cancer patients.

Example 113: in vitro antiproliferative activity

In a cell proliferation assay, the effect of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on inhibiting cell proliferation was tested in a cell line derived from human gastric cancer.

Cell culture/growth inhibition assay: human gastric cancer Hs746t cells were obtained from ATCC (Manassas, VA). Hs746t cells were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin (100U/ml) and streptomycin (100. mu.g/ml). At 37 deg.C, 5% CO₂And culturing the cells under 100% humidity. For cell proliferation assays, cells were plated at 3000 cells/100 μ l/well in white 96-well plates with a clear bottom. After 24hr, the cell culture medium was removed and replaced with media containing various doses of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. After 48hr incubation at 37 ℃, ATP levels were determined using CellTiterGlo (Promega, Madison, Wis.), and luminescence was read using LJL Biosystems analysis HT (Sunnyvale, Calif.). ATP levels were determined in triplicate using independent wells for each dose.

Relative cell number= (mean RLU treated with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide))/(mean RLU vehicle only control).

Figure 19 is a graph of cell number (relative to vehicle) versus N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide concentration and shows that N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide inhibits proliferation of human gastric cancer Hs746t cells after 48 hours of treatment.

Example 114: in vitro antiproliferative activity

The effect of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on inhibiting cell proliferation was tested in cell lines derived from human gastric adenocarcinoma ("gastric carcinoma") in a cell proliferation assay.

Cell culture/growth inhibition assay

Human gastric adenocarcinoma AGS cells were obtained from ATCC (Manassas, VA). AGS cells were cultured in DMEM/F12 supplemented with 10% fetal bovine serum, penicillin (100U/ml) and streptomycin (100. mu.g/ml). At 37 deg.C, 5% CO₂And culturing the cells under 100% humidity. For cell proliferation assays, cells were plated at 3000 cells/100 μ l/well in white 96-well plates with a clear bottom. After 24hr, the cell culture medium was removed and replaced with media containing various doses of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide. After 3 days of incubation at 37 ℃, ATP levels were determined using CellTiterGlo (Promega, Madison, WI) and luminescence values were read using LJL biosystems analysis HT (Sunnyvale, CA). ATP levels were determined in triplicate using independent wells for each dose. In another experiment, 1000 cells/100 ul/well were plated and cells were treated for 6 days and assayed as above.

FIGS. 15A and 15B are graphs showing the concentration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide versus cell number (relative to vehicle) after 3 days of contact with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (A) and after 6 days of contact with (B), indicating that N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide inhibits proliferation of human gastric adenocarcinoma AGS cell lines.

Example 115: growth response of in situ human Hep3B tumor in nude mice treated with varying amounts of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

The dose response potency of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide ("compound a") to inhibit in situ hep3b2.1-7 human hepatoma development was evaluated in BALB/c nu/nu mice in comparison to the optimal dose (75mg/kg) of 5-fluorouracil.

Animals: female BALB/c nu/nu mice (University of Adelaide, white Campus, SA, Australia), 10-14 weeks of age, body weight range: 19.1-29.94g (22.95 g on average) were used for the study. Mice were divided into 6 study groups (4 treatment groups and 2 control groups) as follows:

number of mice/group: including 10 in groups 1-5

15 of the "absorption Rate" control group (group 6)

Mice were kept in a controlled environment (target range: temperature 21 + -3 ℃, humidity 30-70%, 10-15 air changes/hour) under barrier (quarantine) conditions with a 12 hour light/12 hour dark cycle. The temperature and relative humidity are continuously monitored. Commercial rodent chow (Rat and Mouse Cubes, Speciality Feeds PtyLtd, Glen Forrest, Western Australia) and tap water were supplied to animals ad libitum. Both food and water supplies were sterilized by autoclaving.

Tumor inoculation: hep3B human hepatoma cells (passage 2 from working Stock VP-Stock 353) were cultured in RPMI1640 cell culture medium supplemented with 10% FBS and penicillin-Streptomycin (50IU/mL final concentration). Cells were harvested by trypsinization, washed twice in HBSS, and counted. The cells were then resuspended in HBSS: Matrigel (1:1, v/v) and adjusted to contain 1X10 ⁸Final volume of cells/mL. Prior to inoculation, the incision site was thoroughly wiped with alcohol and the abdominal wall was incised to expose the liver. The needle was inserted into the surface of the liver where 10. mu.L of cells (1X 10) were injected⁶A cell). To avoid leakage of tumor cells into the abdominal cavity, the needle was held in this position for about 30 seconds to allowAnd (4) polymerizing.

Treatment was started 14 days after inoculation. On study day 7 (day 21 post-inoculation), all mice from the "uptake" control group were sacrificed and the livers were visually evaluated to detect the presence of tumors.

Materials: the following materials were obtained from various suppliers.

Sterile saline solution (0.9% NaCl (aq)). was obtained from Baxter Healthcare Australia, Old Toongabobie, NSW, Australia. Milk float EL was obtained from Sigma-Aldrich Pty Ltd, Castle Hill, NSW, Australia. A clear colorless liquid clinical preparation of 5-fluorouracil was obtained from Mayne Pharma Pty Ltd. RPMI1640 cell culture medium, FBS and HBSS were obtained from Invitrogen, Pty Ltd, Mt Waverley, VIC, Australia. Penicillin-streptomycin and trypan blue were obtained from Sigma-Aldrich, Castle Hill, NSW, australia. Hep3B2.1-7 human hepatoma cells were derived from the American Type Culture Collection (ATCC), Rockville, Md., USA. Matrigel Obtained from BD Biosciences, North Ryde, NSW, australia.

Use in inoculation suspensionsIncrease the uptake rate of the tumor and reduce the variability of the tumor size,and the growth of Hep3B2.1-7 human hepatoma cells was more stable when seeded in the presence of this extracellular matrix.

Compound formulation and administration: cream flons were dosed as saline (1:9, v/v; vehicle control), N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide ("compound a") or 5-fluorouracil (compound control) according to the following schedule:

vehicle control milk floes/saline (1:9, v/v) was administered orally once daily for 21 days (day 0 to day 20) at a dosing volume of 10 mL/kg.

N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was formulated in cream suspension EL saline (1:9, v/v). Stock solutions were prepared weekly and stored at 4 ℃. Dosing solutions were prepared each day of dosing. N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was orally administered once daily for 21 days (day 0 to day 20) at an administration volume of 10 mL/kg. The compounds were administered at doses of 2, 10 and 50 mg/kg.

The 5-fluorouracil clinical formulation was diluted in sterile saline and administered intravenously via the tail vein at a concentration of 75mg/kg at a dosing volume of 10mL/kg once a week for 3 weeks (on days 0, 7 and 14).

No treatment was applied to mice in group 6 ("uptake rate" control). On study day 7 (21 days post inoculation), mice were sacrificed and livers were exposed to determine "uptake rate" and tumor size within the liver wall.

The body weight of each animal was measured immediately before dosing. The dose to each mouse was calculated and adjusted based on body weight.

Tumor measurement: liver and tumor wet weights were measured when each liver and tumor was excised post-mortem on the day of study termination. At the end of the study, livers were excised from all mice of each study group and weighed. If visible tumors are present, the number is counted. These tumors were removed from the liver and weighed.

Data measurement and sample collection schedules

Data acquisition and calculation: immediately prior to Data acquisition, the transponder of each animal (Bar Code Data Systems Pty Ltd, botanybay, NSW) was scanned using a barcode reader (LabMax I, DataMars, Switzerland). All measurements were taken with the same hand-held caliper (Absolute digital Model CD-6 "CS, Mitutoyo Corporation, Japan). Using Pendra Forms 4.0: ( Software Corporation, Libertyville, IL, u.s.a.) as the transport Software, data was synchronized using the vivo pharm's security relationship database. Aidamm v2.4 was used for data reporting and data calculation.

Counting and calculating: all statistical calculations were performed using SigmaStat 3.0 (SPSS australia Pty Ltd, North Sydney, NSW, australia).

A two-sample t-test was used to determine the significance of changes in body weight in the treated groups between study day 0 and the day of termination. In the case that the data did not pass the normality test or the homogeneity of variance test, the Mann-Whitney rank sum test was performed.

At the end of the study, one-way analysis of variance (ANOVA) was performed on the liver weight and tumor weight data (All pair-wise Multiple Comparison method (All pair Multiple Comparison Procedure) and Multiple Comparison to control group). In the case where the test did not pass the homogeneity of variance test, Kruskal-Wallis one-way analysis of variance (ANOVA) was performed by rank. The same statistical analysis was performed on data from tumor-bearing mice in the study.

P values below 0.05 are considered significant.

Liver weight and tumor weight data of tumor-bearing mice and average weight of liver and tumor of each mouse of each group of tumor-bearing mice

Samples were not taken from mice in group 5 (75mg/kg 5-fluorouracil), which were sacrificed during the study. The study was terminated 18 days after the initial treatment due to the presence of large tumors in some mice (as indicated by the appearance of abdominal swelling).

The dose-dependent trend of liver and tumor weight reduction was evident in the N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide-treated group. When only tumor-bearing mice were considered, the average weight of liver was found to be significantly different in the group treated with the highest dose (group 4 at 50mg/kg) of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide and 5-fluorouracil (group 5 at 75mg/kg) relative to the vehicle control group (group 1; p < 0.05). Furthermore, the mean weights of tumors in the N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (10 mg/kg and 50mg/kg for groups 3 and 4, respectively) and 5-fluorouracil (75mg/kg for group 5) treated groups were found to be significantly different relative to the vehicle control group.

These results are given graphically in figure 16 (mean liver weight-tumor-only mice) and figure 17 (liver tumor weight-tumor-only mice).

Example 116: growth response of orthotopic human HT-29 colon tumor in nude mice treated with varying amounts of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide

The dose-responsive potency of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide ("Compound A") for inhibition of in situ HT-29 human colorectal adenocarcinoma development was evaluated in BALB/c nu/nu mice in comparison to the optimal dose of 5-fluorouracil (75 mg/kg).

Animals: female BALB/c nu/nu mice (University of Adelaide, white Campus, SA, Australia), 7-12 weeks of age, body weight range: 16.58-25.39g (21.52 g on average) were used for the study. Mice were divided into 6 study groups (4 treatment groups and 2 control groups) as follows:

number of mice/group: including 10 in groups 1-5

9 out of the "absorption" control group (group 6)

Tumor inoculation: HT-29 human colorectal adenocarcinoma cells (passage 4 from working Stock VP-Stock 325) were cultured in RPMI1640 cell culture medium supplemented with 10% FBS and penicillin-streptomycin (50IU/mL final concentration). Cells were harvested by trypsinization, washed twice in HBSS, and counted. The cells were then resuspended in HBSS and adjusted to contain 2x10⁸Final volume of cells/mL. Prior to inoculation, the incision site was thoroughly wiped with alcohol and the abdominal wall was incised to expose the cecal wall. The needle was inserted into the surface of the cecum wall where 5 μ L of cells (1X 10) were injected⁶A cell).

Materials: the following materials were obtained from various suppliers.

Sterile saline solution (0.9% NaCl (aq)). was obtained from Baxter Healthcare Australia, Old Toongabobie, NSW, Australia. Milk float EL was obtained from Sigma-Aldrich Pty Ltd, Castle Hill, NSW, Australia. A clear colorless liquid clinical preparation of 5-fluorouracil was obtained from Mayne Pharma Pty Ltd. RPMI1640 cell culture medium, FBS and HBSS were obtained from Invitrogen, Pty Ltd, Mt Waverley, VIC, Australia. Penicillin-streptomycin and trypan blue were obtained from Sigma-Aldrich, Castle Hill, NSW, australia. HT-29 human colorectal adenocarcinoma cells were derived from the American Type Culture Collection (ATCC), Rockville, Md., USA.

Compound formulation and administration: cream flons were dosed as saline (1:9, v/v; vehicle control), N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, or 5-fluorouracil (compound control) according to the following schedule:

N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was formulated in cream suspension EL saline (1:9, v/v). Stock solutions were prepared weekly and stored at 4 ℃. Dosing solutions were prepared each day of dosing. N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is administered orally once daily for 21 days (day 0 to day 20) at doses of 2, 10 and 50mL/kg, in an administration volume of 10 mL/kg.

No treatment was applied to mice in group 6 ("uptake rate" control). On study day 7 (21 days post inoculation), mice were sacrificed and the colon was exposed to determine "uptake rate" and tumor size within the blinded intestinal wall.

Tumor measurement: the cecal and tumor wet weights were measured when each cecal and tumor was excised post-mortem on the day of study termination. At the end of the study, ceca were excised from all mice of each study group and weighed intact with the tumor. Tumors were then excised from the cecum and weighed.

Data measurement and sample collection schedules

Data acquisition and calculation: immediately prior to Data acquisition, the transponder of each animal (Bar Code Data Systems Pty Ltd, botanybay, NSW) was scanned using a barcode reader (LabMax I, DataMars, Switzerland). All measurements were taken with the same hand-held caliper (Absolute digital Model CD-6 "CS, Mitutoyo Corporation, Japan). Using Pendra Forms 4.0: (Software Corporation,LibertyvilleIL, u.s.a.) as transport software, data is synchronized using the vivo pharm's security relationship database. Using AIDAM v2.4 for data reporting and data computation

A two-sample t-test was used to determine the significance of changes in body weight in the treated groups between study day 0 and the day of termination. In the group treated with 2 and 50mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, the treatment was discontinued due to excessive weight loss. In these groups, a two-sample t-test was used to determine the significance of body weight changes in the treatment groups between day 0 of the study to the final treatment day and between the last treatment day of the study to the termination day. In the case that the data did not pass the normality test or the homogeneity of variance test, the Mann-Whitney rank sum test was performed.

At the end of the study, single-factor analysis of variance (ANOVA) was performed on the cecal and tumor weight data (All pair-wise Multiple Comparison (All pair Multiple Comparison Procedure) and Multiple Comparison to control). In the case where the data does not pass the normality test, the values are converted to natural logarithms before this method is performed. .

P values below 0.05 are considered significant.

And (4) observation: mean body weight loss was measured in all study groups, including vehicle control group. In all study groups, including vehicle control, signs of diarrhea and dehydration (loss of skin elasticity) were observed. Severe weight loss early during the study period resulted in discontinuation of treatment in the group receiving the lowest dose (2mg/kg) and highest dose (50mg/kg) of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide on days 9 and 7 of the study, respectively. Since the body weight loss was less severe in the group receiving 10mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, all treatments were administered to this group according to the schedule. For this group and the 5-fluorouracil treated group, mean body weight loss was significant at the end of the study.

Although the uptake rate of HT-29 tumors was 100% in the "uptake rate" group 21 days after inoculation, the size of these tumors was much lower than expected. This may result in no significant difference in mean cecal and tumor weight between the N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide treated group and the vehicle control group. 5-Fluorouracil also had no effect on the weight of cecum and HT-29 tumors.

Body weight measurement (± SEM) (final treatment day and study end date)

Body weight data was not collected for group 6 ("absorption" control). This group of mice was sacrificed at day 7 of the study (day 21 post-inoculation) to visually assess whether tumor growth was sufficient for the purposes of this study.

Treatment in group 2 (2mg/kg N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide) was discontinued on study day 9 and treatment in group 4 (50mg/kg N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide) was discontinued on study day 7 due to excessive weight loss by the mice. The remaining groups received all regular treatments during the study.

The mean tumor weight for each group is shown in figure 18. The mean tumor weight for each group included only mice that survived to the final day of the study. Values for mice that died during the study were not included in the calculated mean.

Cecal and tumor weight data

Shaded boxes represent samples taken from mice that died during the study. The calculated average of the cecal and tumor weights did not include these values. The trend shows that data for HT-29 tumor and cecal weight are reduced following treatment with 10mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Example 117: tumor growth delay in nude mice bearing human A375 melanoma xenografts

6 groups (n =9) of tumor-bearing mice were used. The control group included one mouse receiving 10% creosote EL/saline vehicle once a day for 14 days (qdx14) by oral gavage (po) and another mouse given paclitaxel as a reference agent by tail vein injection (iv) at 30mg/kg for 5 times every other day (qod x 5). 4 test groups received 25mg/kg or 50mg/kg, qd x14, or 12.5 or 25mg/kg, bid x14 of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide ("Compound A") orally. Treatment outcome was assessed by TGD, which is defined as the difference in median time to endpoint tumor volume in the treated group compared to the control group. Toxicity was assessed by body weight measurements and clinical observations.

Animals: on day 1 of the study, female athymic nude mice (nu/nu, Harlan) were 10-11 weeks old and had Body Weights (BW) ranging from 19.3 to 19.325.5 g. These animals were fed water ad libitum (reverse osmosis, 1ppm Cl) and NIH 31 modified and Irradiated LabThe feed consisted of 18.0% crude protein, 5.0% crude fat and 5.0% crude fiber. In a stationary micro-insulated incubator (microisolator) under a 12 hour photoperiod at 21-22 deg.C (70-72 deg.F) and 40-60% humidity And (5) feeding the mice on the experimental animal padding. The recommendations for restrictions, animal feeding management, surgical procedures, feed and liquid provisions, and veterinary Care were followed with guidelines for the management and Use of Laboratory Animals.

Tumor implantation: xenografts were initiated from a375 human melanoma tumors by serial transplantation in athymic nude mice. A375 tumor fragments (. about.1 mm)³) Subcutaneously implanted in the right flank of each test mouse and when the average size was close to 100-³Tumor growth was monitored. After 13 days, designated as study day 1, animals were divided into 6 groups, each group ranging from 63 to 221mm in tumor volume³And the group mean tumor volume is 125.3 to 125.9mm³9 mice (decreasing from 10) were composed. Tumor volume was calculated using the formula:where w = the width (mm) of the a375 tumor, and l = the length (mm) of the a375 tumor.

Materials: n- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide was dissolved in 10% cream EL in saline at a concentration of 5mg/mL, with sonication, shaking and heating to 35 ℃ to aid dissolution. 5mg/mL solution was used as the 50mg/kg therapeutic dosing solution and dosing solutions for 25mg/kg and 12.5mg/kg treatments were prepared by serial dilution. The dosing solution was stored up to 1 week at room temperature in the dark.

A daily dosing solution of paclitaxel (NPI) was prepared by diluting 30mg/mL stock solution to 3mg/mL in 5% ethanol, 5% cream EL in 5% aqueous glucose (D5W). The paclitaxel dose is 30 mg/kg.

Treatment: the following table shows the treatment regimen.

Mice in group 1 received 14 daily doses (qd x14) of vehicle consisting of 10% milk buoyant EL in saline by oral gavage (po) and served as controls for tumor development. Group 2 animals were dosed intravenously (iv) with paclitaxel as a reference reagent at 30mg/kg, 5 doses every other day (qod x 5). Groups 3-6 mice received oral N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide according to the following schedules: 50mg/kg, qd x 14; 25mg/kg, twice daily, 14 days, single dose (bidx14) on day 1 and on the last 1 day; 25mg/kg, qd x 14; and 12.5mg/kg, bid x 14. All doses were administered at a volume of 0.2 mL/20 g body weight and adjusted to the body weight of the animals.

End point: tumors in all groups were measured twice weekly using calipers. When the tumor size of each animal reached the end point of 2000mm³Either first, or on the final day of the study (day 60), each animal was euthanized. The Time To Endpoint (TTE) was calculated for each mouse from the following equation: Where b is the intercept of a straight line and m is the slope of the straight line obtained by linear regression of log transformed tumor growth data sets.

The data set included a first observation beyond the study endpoint volume and three consecutive observations immediately prior to reaching the endpoint volume. Animals that did not reach the endpoint were assigned a TTE value equal to the last day of the study. Animals classified as NTR (non-treatment related) deaths due to accident (NTRa) or due to unknown cause (NTRu) were not included in the TTE calculation (and all further analyses). The TTE value for animals assigned to TR (treatment-related) death or NTRm (non-treatment-related due to metastasis) death is equal to the day of death.

Treatment outcome was determined from Tumor Growth Delay (TGD), which is defined as the increase in median Time To Endpoint (TTE) in the treated group compared to the control group: TGD = T-C, expressed in days, or as a percentage of the median TTE of the control group:wherein: t = median TTE for treatment group, C = median TTE for control group (group 1).

Treatment may cause Partial Regression (PR) or Complete Regression (CR) of the tumor in the animal. In the PR response, the tumor volume measured in three consecutive measurements was 50% or less of the day 1 volume during the study, and the tumor volume measured in one or more of these 3 measurements was equal to or greater than 13.5mm ³. In the CR response, three consecutive measurements of tumor volume were less than 13.5mm during the study³. At the end of the study, animals with CR response were additionally classified as tumor-free survivors (TFS). Tumor regression was monitored and recorded.

Side effects: animals were weighed daily for the first 5 days of the study, and twice weekly thereafter. Mice are frequently observed for significant signs of any adverse, treatment-related side effects, and clinical signs are recorded when observed. Acceptable tolerability was defined as a mean weight loss of less than 20% of the group's body weight during the test period and no more than 1 treatment-related death in the animal group. Any dosing regimen that does not meet these criteria is considered to exceed the Maximum Tolerated Dose (MTD). Death is classified as TR if clinically indicated and/or autopsied to be attributable to a side effect of the treatment, or may be classified as TR if due to unknown causes during administration or within 10 days of the last administration. If there is no evidence of death associated with side effects of treatment, death is classified as NTR.

Statistical and graphical analysis: the significance of the difference between the TTE values of the treated and control groups was analyzed using a time series test. At significance level P =0.05, two-tailed statistical analysis was performed.

The median tumor growth curve indicates the median tumor volume of the group as a function of time. When animals were withdrawn from the study due to tumor size or TR death, the final tumor volumes of the animals recorded and the data were included for calculation of median group tumor volumes at subsequent time points. The curve was truncated after 50% of the animals in the group exited the study due to tumor progression. Kaplan-Meier plots were made to represent the percentage of animals remaining in the study as a function of time, and the same data set was used as for the time series test. Prism (graphpad) Windows 3.03 was used for all graphical and statistical analyses.

Summary of treatment response

Growth of a375 tumors in control mice (group 1): animals in group 1 received 10% milk buoyant EL/saline vehicle, po, qd x 14. Tumors of control mice grew progressively to 2000mm³The endpoint volume, median TTE was 22.8 days (the maximum possible T-C was determined in the 37.1 day study) or the TGD was 163%.

Effect of treatment with paclitaxel (group 2): paclitaxel, 30mg/kg, iv, qod x5, was administered as a reference agent to animal 2. All 9 animals reached the tumor volume endpoint. Tumor growth was parallel and slightly shifted to the right compared to control. The median TTE of 28.8 days, corresponding to 26% TGD, was significant by time series analysis (table 2, P = 0.0088G 1 vs. G2). No tumor regression was associated with paclitaxel treatment.

Effect of treatment with N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide (groups 3-6): groups 3-6 received oral administration of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide as monotherapy. Group 3 animals were dosed with 50mg/kg on the qdx14 schedule. The 9 tumors in the group reached a volume endpoint. The median tumor volume of the groups experienced a slight net change between days 1-10, and then continued to increase with the study. One animal underwent tumor PR. The median TTE was 27.5 days, or 21% TGD, as a significant result (P = 0.0054G 1 vs.g).

Animals of group 4 received 25mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide according to the bid x14 schedule. On day 60, 4 of the 9 animals in the group remained, all being TFS. The day before the end of the study, tumors of additional 2/9 animals reached a volume endpoint. This group had 4/9PR, 5/9CR and 4/9 TFS. On the first few days of the study, the median tumor volume began to decrease and lasted for about 30 days. 5/9 tumor regrowth in animals resulted in a recurrence of the median tumor growth starting at about day 32 and continuing until the end of the study. The median TTE for this group was 59.9 days, representing the maximum possible 163% TGD (P <0.0001, table a 1).

Group 5 mice also received 25mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, but according to the less stressful qd x14 schedule. All 9 animals in group 5 reached the tumor volume endpoint with no tumor regression. Tumor growth varied closely along the tumor growth trajectory of the control group. Median TTE of 25.6 days, or 12% TGD, was a non-significant result (P = 0.0662G 1 vs. G5).

Group 6 animals were dosed with 12.5mg/kg of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide according to the bid x14 schedule. All tumors in the group reached a volume endpoint. As with group 4, the median tumor volume decreased in group 6 early in the study, but this decrease lasted only about 9 days and was associated with a single PR response. Tumor volume increased from day 10 to the end of the study. The median TTE for this group was 27.5 days, corresponding to 21% TGD with significance (P = 0.0424G 1 vs. G6).

In summary, N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide, administered orally once daily and twice daily, showed dose-dependent antitumor activity against human a375 melanoma xenografts. Twice daily dosing was preferred over once daily dosing in terms of the magnitude of TGD produced and the number of responses targeted. Thus, the antitumor activity of N- (S) - (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide is both dose-and time-table dependent.

Example 118: inhibiting activity of subcutaneous COLO 205 human colon cancer xenografts

Animals: on day 1 of the study, female athymic nude mice (nu/nu, Harlan) were 12-13 weeks old and had Body Weights (BW) ranging from 18.3 to 27.3 g. These animals were fed water ad libitum (reverse osmosis, 1ppm Cl) and NIH 31 modified and Irradiated LabThe feed consisted of 18.0% crude protein, 5.0% crude fat and 5.0% crude fiber. Irradiating in a static micro-insulated incubator at 21-22 deg.C (70-72 deg.F) and 40-60% humidity under a 12-hour photoperiodAnd (5) feeding the mice on the experimental animal padding. Recommendations for restrictions, animal feeding management, surgical procedures, feed and liquid provisions, and veterinary care were followed with the guidelines for laboratory animal management and use.

Tumor implantation:xenografts were initiated from COLO 205 human colon cancer cells. Tumor cells were cultured in 10% heat-inactivated fetal bovine serum, 100 units/mL penicillin G sodium, 100. mu.g/mL streptomycin sulfate, 0.25. mu.g/mL amphotericin B and 25. mu.g/mL gentamicin, 2mM glutamine, 1mM sodium pyruvate, 10mM HEPES and 0.075% sodium bicarbonate. At 37 ℃ in 5% CO₂And 95% air in a humidified incubator, cells were cultured in tissue culture flasks. Colo 205 cells were collected during logarithmic growth on the day of tumor cell implantation and at 5x10 ⁶The concentration of cells/mL was resuspended in 50% Matrigel matrix (BD Biosciences) in PBS. Each test mouse received a subcutaneously implanted 1X10 in the right flank⁶Colo 205 cells, and when the average size is close to 80 to 120mm³Tumor growth was monitored. After 14 days, designated day 1 of the study, animals were divided into 8 groups (n =9) with tumor volumes ranging from 63 to 196mm³And the group mean tumor volume is 118 to 119mm³. Tumor volume was calculated using the formula:where w = width (mm) of COLO 205 tumor, and l = length (mm) of COLO 205 tumor. Assuming that 1mg corresponds to 1mm³The tumor weight can be estimated.

Materials: fresh solutions for compound a administration were prepared daily by dissolving the required amount of compound in 100% cream EL, and then diluted 10-fold with physiological saline. To provide a dose of 25, 50, 100 or 200mg/kg in a 10mL/kg dosing volume, respectively, the final dosing solution concentration is 2.5, 5, 10 or 20 mg/mL. Paclitaxel was formulated fresh each day of administration in a vehicle consisting of 5% ethanol and 5% cream EL in 90% D5W (5% EC vehicle) (Natural Pharmaceuticals, Inc.).

Treatment: the following table shows the treatment regimen.

Group 1 received the formulation vehicle (10% cream EL in saline) and served as the tumor growth control group. Group 2 received the reference drug paclitaxel administered according to its optimal schedule in nude mice (30mg/kg i.v. qod x 5). Groups 3-6 received compound a at doses of 25, 50, 100 and 200mg/kg, respectively, administered at p.o.qd x14, and administration of group 6 (200mg/kg) was discontinued after 6 days due to toxicity. All doses (0.2mL/20g body weight) were adjusted according to the weight of the animals.

End point: tumors were measured twice weekly using calipers. When the tumor size of each animal reached the predetermined endpoint size of 2000mm³Either first, or on the final day of the study (day 74), each animal was euthanized. However, up to about 800mm in size³Control tumors did not show log growth characteristics. Thus, the end-point tumor size is 800mm³For analysis of Tumor Growth Delay (TGD). The Time To Endpoint (TTE) was calculated for each mouse from the following equation:where b is the intercept of a straight line and m is the slope of the straight line obtained by linear regression of log transformed tumor growth data sets. The data set included a first observation beyond the study endpoint volume and three consecutive observations immediately prior to reaching the endpoint volume. TTE values for animals not reaching the endpoint were assigned to be equal to the last day of the study. Animals classified as NTR (non-treatment related) deaths due to accident (NTRa) or due to unknown cause (NTRu) were not included in the TTE calculation (and all further analyses). The TTE value for animals assigned to TR (treatment-related) death or NTRm (non-treatment-related due to metastasis) death is equal to the day of death.

Treatment outcome was evaluated by Tumor Growth Delay (TGD), which is defined as the increase in median Time To Endpoint (TTE) in the treated group compared to the control group: TGD = T-C, expressed in days, or as a percentage of the median TTE of the control group: Wherein: t = median TTE for treatment group, C = median TTE for control group.

The control group was designated as group 1 mice.

Treatment may cause Partial Regression (PR) or Complete Regression (CR) of the tumor in the animal. In the PR response, the tumor volume measured in three consecutive measurements was 50% or less of the day 1 volume during the study, and the tumor volume measured in one or more of these 3 measurements was equal to or greater than 13.5mm³. In the CR response, three consecutive measurements of tumor volume were less than 13.5mm during the study³. The degradation response is monitored and recorded.

Side effects: animals were weighed daily for the first 5 days of the study, and twice weekly thereafter. Mice are frequently observed for significant signs of any adverse, treatment-related side effects, and clinical signs of toxicity are recorded when observed. Acceptable toxicity is defined as a group average weight loss of less than 20% over the study period and no more than 1 treatment-related (TR) death in 10 treated animals, any dosing regimen that resulted in greater toxicity being considered beyond the Maximum Tolerated Dose (MTD). Death is classified as TR if clinically indicated and/or necropsied as attributable to a side effect of the treatment, or can be assessed as TR if due to unknown causes during the administration or within 10 days of the last administration. If there is no evidence of death associated with side effects of treatment, death is classified as NTR. Animals were monitored for side effects by frequent observation and BW measurements. BW changes were insignificant and all treatments were acceptably tolerated except group 6. Once daily, 6 p.o. doses of 200mg/kg compound a resulted in 1 TR death on day 7 of the evaluation and another 2 TR deaths on day 8. All mice in group 6 showed clinical signs of toxicity, including a raised posture, hypoactivity, and loose stools.

The median tumor growth curve indicates the median tumor volume of the group on a logarithmic scale as a function of time. When animals were withdrawn from the study due to tumor size or TR death, the final tumor volumes of the animals included the recordings and the data used to calculate the median group tumor volumes at subsequent time points. The curve was cut off after 50% of the animals in the group had withdrawn from the study due to tumor progression, or after the 2 nd TR in the group. Kaplan-Meier plots were made to show the percentage of animals remaining in the study as a function of time, and the same data set was used as for the time series test. Prism (graphpad) Windows 3.03 was used for all graphical and statistical analyses.

Summary of treatment response

Growth of COLO 205 tumors in control mice (group 1)

Tumors of group 1 showed slow heterosexual growth. 7/9 vehicle-treated group 1 control mice had tumors reaching 800mm³Tumor volume endpoint, and two mice remained at the end of the study. The median TTE for group 1 was 41.0 days and thus the maximum possible TGD in this 74-day study was 33.0 days (80%).

Effect of treatment with paclitaxel (group 2)

On day 74, 8 group 2 mice (n =9) with MTV143mm were left in the study receiving paclitaxel treatment (day 9)³. This corresponds to the maximum possible TGD (33.0 days or 80%) and statistically significant activity (P = 0.002). 5 PR responses were recorded. The median tumor growth curve indicates a decrease in MTV up to day 19, followed by a few changes until day 47 when tumor growth is restored.

Effect of treatment with Compound A (groups 3-6)

The resulting TTE median values for groups 3, 4, and 5 were 47.9, 59.1, and 74.0 days, respectively. Groups 3 and 4 had non-significant temporal verification results, and group 5 temporal verification reached edge significance (P = 0.058). These treatments produced a dose-dependent number of degenerations, however, the type of degenerating response (PR vs. cr) and the number of 74-day survivors per group were not dose-related. The median tumor growth curve indicates that at the early stages of the study (up to day 29), the activity was similar for 3 dose levels, followed by a dose-dependent delay in tumor regeneration. Group 6 produced 3 TR deaths and dosing was discontinued after day 6. Therefore, 200mg/kg treatment was considered to exceed MTD and TGD could not be evaluated.

Compound a showed dose-dependent activity against COLO 205 colon cancer xenografts. When administered at 25mg/kg, compound a showed 3% TGD. At 50mg/kg, Compound A produced 46% TGD. Acceptably tolerating 100mg/kg treatment and, like paclitaxel treatment, resulted in the largest possible TGD in the trial, with a similar number of degenerative responses. Treatment at 200mg/kg produced 3/9 TR deaths and exceeded the MTD. A more significant initial reduction in tumor burden was observed for compound a compared to paclitaxel; however, the duration of the effect is short. In comparison to the control, in the 25 and 50mg/kg groups, initial tumor regeneration progressed more rapidly, and by the end of the study, the MTV was close to that of the control. The 100mg/kg treatment did not show this rapid regeneration, but did show faster tumor growth compared to the tumor regeneration of paclitaxel.

Example 119: human clinical trial

Random, double-blind, label-published, history-controlled, single-cohort safety/efficacy human phase I clinical trials will be performed with compound a-controlled placebo in patients with chemotherapy-naive (chemi-naive) advanced or metastatic pancreatic cancer.

The primary objective of this study was to evaluate the safety and tolerability of compound a. Secondary results were to assess response rate, clinical benefit and tumor shrinkage after treatment with compound a. In addition, the study will be designed to assess the time to disease progression and overall survival of pancreatic cancer patients. Furthermore, pharmacodynamic changes in tumor vascular parameters (including, e.g., blood flow, blood volume, time to peak of the ROC subject operating characteristic curve) were evaluated by DCE-MRI.

Furthermore, biological markers such as MEK1 and MEK2 genetic polymorphisms and serum proteomics will be used to correlate the results. This would also allow the determination of tumor resection rate after treatment and the evaluation of compound a MTD.

Compound a was administered at different doses during the study, which were: about 1mg, about 1.5mg, about 2mg, about 2.5mg, about 3mg, about 3.5mg, about 4.0mg, about 4.5mg, about 5mg, about 5.5mg, about 6mg, about 6.5mg, about 7mg, about 7.5mg, about 8mg, about 8.5mg, about 9mg, about 9.5mg, about 10mg, about 10.5mg, about 11mg, about 11.5mg, about 12mg, about 12.5mg, about 13mg, about 13.5mg, about 14mg, about 14.5, or about 15 mg.

The selection criteria for this study will be based on the following factors:

● locally advanced unresectable or marginally unresectable pancreatic cancer confirmed histologically/pathologically, and no evidence of metastatic disease.

● diagnosis of locally advanced, non-resectable pancreatic cancer based on assessment by biphasic CT scanning and/or by Endoscopic Ultrasound (EUS), as described in appendix F.

● disease measurable according to RECIST and results obtained by biphasic CT scans within 14 days prior to protocol treatment enrollment.

● the size of the tumor is greater than or equal to 2cm by two-phase computed tomography.

● within 14 days of enrollment, proper organ function is documented by the following evidence: absolute neutrophil count>1500/mm 3; platelet count 100,000/mm 3; hemoglobin³9gm/dL, no transfusion is needed in the first 4 weeks; total bilirubin is less than or equal to 1.5 times of the normal upper limit value (ULN); transaminase (AST and/or ALT) is less than or equal to 2.5x ULN; PT (or INR) ≦ 1.5x ULN andaPTT within normal limits (patients receiving anticoagulant therapy with agents such as warfarin or heparin will be admitted; for patients receiving warfarin, as defined by local standard of care, at least weekly assessments are closely monitored until INR is stable based on measurements at the previous dose; creatinine clearance calculated using the Cockcroft-Gault equation >60ml/min。

Exclusion criteria included: pre-treatment with compound a within 6 months prior to enrollment; clinical evidence of tumor invasion of the duodenal mucosa (as documented by endoscopy or sonoendoscopy); minor surgery (such as fine needle aspiration biopsy or needle aspiration biopsy) within 14 days of study enrollment; major surgery, significant traumatic injury, or severe non-healing wounds, ulcers or fractures within 21 days of study enrollment; within 6 months prior to study drug administration, any of the following: severe/unstable angina (angina symptom at rest), new onset angina (starting within the last 3 months) or myocardial infarction, congestive heart failure, cardiac ventricular arrhythmias requiring anti-arrhythmic therapy; over the past 6 months, a history of thrombotic or embolic events such as cerebrovascular accidents or transient ischemic attacks; a history of aneurysm or arteriovenous malformation; known Human Immunodeficiency Virus (HIV) infection or chronic hepatitis B or chronic hepatitis C; clinically severe infections with activity above CTCAE grade 2; within 4 weeks of study enrollment, receiving any study medication; uncontrolled hypertension, defined as systolic blood pressure greater than 150mmHg or diastolic blood pressure greater than 90mmHg, despite optimal medical management; pulmonary blood loss/bleeding events above CTCAE grade 2 within 4 weeks of study enrollment; any other blood loss/bleeding events above CTCAE grade 3 within 4 weeks of study enrollment; a demonstration or history of hemorrhagic diathesis or coagulopathy; chronic daily treatment with aspirin or other non-steroidal anti-inflammatory drugs; using st.john's Wort, rifampicin, ketoconazole, itraconazole, ritonavir or grapefruit juice; known or suspected to be allergic to compound a; any condition that impedes the ability of the patient to swallow the entire pill; any malabsorption problems; other severe acute or chronic medical or psychological conditions, or laboratory abnormalities that may increase the risk associated with the administration of substances or drugs involved in a study, or that may interfere with the interpretation of the results of a study, and which, at the investigator's discretion, would render a patient unsuitable for participation in the study; a history of collagen vascular disease; any contraindications for magnetic resonance imaging are accepted.

Example 120: human clinical trial

A phase I human clinical trial of safety/efficacy was performed with compound a in patients with advanced or metastatic gastric cancer who received chemotherapy for the first time, randomly, double-blindly, open-label, historically-controlled, single cohort, in the same manner as described in example 117, except that enrolled patients were diagnosed with lymphoma, gastric stromal tumor, or gastric carcinoid tumor.

Example 121: carrageenan-induced paw edema (CPE) in rats

Compound a (6, 20&60mg/kg) or indomethacin (3mg/kg) was administered orally and after 2hr, 1% carrageenan suspension was injected into the right hindpaw of male Sprague-Dawley rats (N = 6/treatment group). After 3hr, hind paw edema was measured by plethysmography to assess paw volume. A reduction of hind paw edema of 30% or more indicates significant acute anti-inflammatory activity. Indomethacin (Indo) was used as a positive control drug. The increase in paw volume in each treatment group is shown in figure 22, indicating that oral administration of compound a caused significant anti-inflammatory activity in the rat carrageenan paw edema model in all dose groups.

Example 122: rat adjuvant arthritis inflammation assay

In the rat adjuvant-induced arthritis model, freund's complete adjuvant (CFA) was injected into the right hind paw of rats to induce a condition similar to rheumatoid arthritis in humans. Compound a was administered orally at 2, 6 and 20mg/kg for 5 consecutive days. In addition, dexamethasone was administered orally at 5mg/kg for 5 days. On days 1 and 4, Enbrel was administered by subcutaneous injection at 10 mg/kg. On day 1, CFA was injected into the right hind paw at 1 hour after the first dose. Inhibition of swelling of the right hind paw was measured on days 1 and 5 relative to vehicle treated controls for the acute phase, and% inhibition of swelling of the left hind paw was measured on days 14 and 18 relative to vehicle treated controls for the delayed phase. Polyarthritis was scored when swelling existed in the forepaw, tail, nose or ear.

The% inhibition of swelling relative to control for the different treatment groups is shown in fig. 23A and 23B. Compound a showed a significant reduction in swelling at 20mg/kg both in the acute and delayed phases. For polyarthritis scores, all 6 animals in the vehicle treated group had swollen forepaws and tails. For compound a group at 20mg/kg, 2 of 6 animals had no swelling in the forepaw and 4 of 6 animals had no swelling in the tail. For the enbrel group, all animals were not protected from paw swelling, and 3 out of 6 animals had no tail swelling.

Example 123: inhibition of Collagen Antibody Induced Arthritis (CAIA) in mice

Male Balb/c mice (N = 8/treatment group) were injected intravenously (tail vein) with 2mg of collagen antibody cocktail (Chondrex) on day 0. On days 0-4, RDEA119(1, 3) was administered orally&10mg/kg QD) or dexamethasone (1mg/kgQD), whereas Enbrel was injected subcutaneously on days 1 and 3. On day 3, the test was carried out except for the first timeAll mice except animals were given an intraperitoneal injection of LPS (50 μ g). Arthritis scores were determined for all limbs and are shown in fig. 24 (highest score 16). Significant anti-inflammatory activity was recorded for all test substances and reference drugs. Enbrel and dexamethasone as positive pairs And (6) irradiating.

Example 124: in vivo cell proliferation assay

Methods for determining the number of cell proliferations in cancer cells treated with MEK protein kinase inhibitors are known in the art and are described in Kenny, L.M. et al, Positron Emission visualization (PET) Imaging of cell Proliferation in Oncology, Clinical Oncology,16:176-185(2004), which is incorporated herein by reference in its entirety. MEK protein kinase inhibitors (e.g. compound a) are tested in vivo to determine their effect on cancer cell proliferation. 50 patients volunteered to the study and all had pancreatic cancer at a similar stage of cancer development. Compound a combinations were administered to 25 patients. Placebo was administered to the last 25 patients. A daily dose is administered to each patient for 14 days in combination with a radiolabeled tracer, such as labeled fluoro-2-deoxy-DF-glucose (FDG).

After 14 days of treatment, trained physicians used a non-invasive Positron Emission Tomography (PET) imager to detect tumor cell proliferation. Furthermore, trained physicians will determine the number of cell proliferations of tumor and normal cell tissues in patients treated with compound a and placebo. The results indicate a reduction in the number of cell proliferations between MEK protein kinase inhibitor (e.g. compound a) and placebo. This assay, which uses labeled tracers and PET imaging to determine the number of cell proliferations, is referred to herein as the "in vivo cell proliferation method". Other methods of in vivo cell proliferation are known in the art.

A similar analysis can be used to determine the reduction in tumor size.

Example 125: in vivo apoptosis assay

MEK inhibitors (e.g., compound a) were tested in vivo to determine their effect on cancer cell apoptosis. 40 patients volunteered to the study and all had pancreatic cancer at a similar stage of cancer development. Compound a was administered to 20 patients and a placebo was administered to 20 patients. The daily dose was administered to each patient for 14 days.

After 14 days, each patient will take up a detectable labeled Lipopolysaccharide Binding Protein (LBP) reagent conjugated. According to WO/2006/054068 (which is incorporated herein by reference in its entirety), each patient is then placed in the area of a scanner which detects the ingested agent in association with dead cells. The number of dead cells can be correlated with the level of apoptosis in each patient. The levels of apoptosis in patients administered the combination and those administered the single entity agent can be compared to each other and to a cohort administered a placebo. This assay using lipopolysaccharide binding proteins and scanners to detect levels of apoptosis is referred to herein as the "in vivo apoptosis method".

Example 126: dissolution study

Capsules containing compound a were prepared as described in the above examples. The following dissolution data were obtained using the USP <711> method for determining dissolution.

	1mg form	10mg form
			Time (min)	% release (% RSD)	% release (% RSD)
15	78(8.3)	80(7.3)
			30	82(7.1)	87(9.2)
45	82(6.7)	92(9.6)
			60	88(6.3)	92(7.2)
70	86(5.7)	95(5.4)

Claims

1. Selected from:use of a compound of (a) in the preparation of a composition, wherein the composition allows for modified release of the compound and is used in the treatment of a MEK mediated disorder, which is brain, lung, ovarian, pancreatic, prostate, renal, colorectal, leukemia, gastric, or liver cancer.

2. The use of claim 1, wherein the lung cancer is non-small cell lung cancer or small cell lung cancer.

3. The use of claim 1, wherein the leukemia is myeloid leukemia, pre-B acute leukemia or chronic lymphocytic B leukemia.

4. The use of claim 1, wherein the liver cancer is hepatocellular carcinoma.

5. The use of claim 1, wherein the brain cancer is glioblastoma.

6. The use of claim 1, wherein the 2-OH carbon on the compound is in the R configuration or in the S configuration.

7. The use of claim 1 or 6, wherein said composition is substantially free of the R-isomer of said compound or substantially free of the S-isomer of said compound.

8. The use of claim 1 or 6, wherein the composition is for use in combination therapy and further comprises another therapeutic agent.

9. The use of claim 8, wherein the other therapeutic agent is sorafenib or gemcitabine and the composition is for the treatment of hepatocellular carcinoma or pancreatic cancer.

10. The use of claim 7, wherein the compound comprises less than 5% of the R-isomer of the compound or less than 5% of the S-isomer of the compound.

11. The use of claim 7, wherein the compound comprises less than 1% of the R-isomer of the compound or less than 1% of the S-isomer of the compound.

12. A composition comprisingOf the polymorph of a shows a powder X-ray diffraction pattern comprising at least 50% of the peaks identified in the powder X-ray diffraction pattern shown in figure 5, and for use in the treatment of a MEK-mediated disorder, which is a brain, breast, lung, ovarian, pancreatic, prostate, renal, colorectal, leukemia, stomach, or liver cancer.

13. The composition of claim 12, wherein the lung cancer is non-small cell lung cancer or small cell lung cancer.

14. The composition of claim 12, wherein the leukemia is myeloid leukemia, pre-B acute leukemia, or chronic lymphocytic B leukemia.

15. The composition of claim 12, wherein the liver cancer is hepatocellular carcinoma.

16. The composition of claim 12, wherein the brain cancer is glioblastoma.

17. The composition of claim 12, wherein said composition is substantially free of the R-isomer corresponding to said polymorph a.

18. The composition of claim 12, wherein the polymorph a is prepared by a process comprising the step of crystallizing amorphous N- (3, 4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

19. The composition of claim 12, wherein the composition is for use in combination therapy and further comprises another therapeutic agent.

20. The composition of claim 19, wherein the other therapeutic agent is sorafenib or gemcitabine and the composition is for the treatment of hepatocellular carcinoma or pancreatic cancer.