TWI481590B - Derivatives of n-(arylamino) sulfonamides including polymorphs as inhibitors of mek as well as compositions, methods of use and methods for preparing the same - Google Patents

Description

N-(arylamino)sulfonamide (including polymorph) derivatives and compositions as MEK inhibitors, use method and preparation method thereof

U.S. Patent Application Serial No. 61/044,886, filed on Apr. 14, 2008, and U.S. Patent Application Serial No. 61/034,466, filed on March 6, 2008, and on March 6, 2008 The priority of the U.S. Patent Provisional Application Serial No. 61/034,464, the entire disclosure of which is incorporated herein by reference.

The oncogene is a mutant of a specific normal cell gene ("primary oncogene"). In certain instances, the oncogene encodes an abnormal form of the signal transduction pathway component, such as a receptor tyrosine kinase, a serine-threonine kinase, or a downstream signal transduction molecule.

A particular embodiment disclosed herein is an inclusion comprising: and A composition of the compound. In certain embodiments, the composition further comprises an excipient selected from the group consisting of microcrystalline cellulose, croscarmellose sodium, sodium lauryl sulfate, magnesium stearate, or a combination thereof. In certain embodiments, the composition comprises from about 0.1% to about 15% by weight Structural compound, A compound of the structure or a combination thereof. In certain embodiments, the composition further comprises from about 85% to about 99.9% by weight microcrystalline cellulose. In certain embodiments, the composition comprises about 1 mg of Structural compound, A compound of the structure or a combination thereof; about 222.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate. In certain embodiments, the composition comprises: about 10 mg Structural compound, a compound of the structure or a combination thereof; about 213.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of the harder embodiment, the combination Contains: about 20mg: Structural compound, A compound of the structure or a combination thereof; about 203.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate. In certain embodiments, the composition comprises: about 40 mg: Structural compound, A compound of the structure or a combination thereof; about 183.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate. In certain embodiments, the compound achieves a _Cmax between about 0.01 [mu]g/ml to about 1.0 [mu]g/ml on Day 1 when administered to an individual. In certain embodiments, the compound has an AUC between about 0.1 μg hour/mL to about 5.0 μg hour/mL from 0 to 12 hours. In certain embodiments, the compound has a plasma concentration of greater than about 0.01 mg/mL after a single dose of about 5 hours.

In a particular embodiment, the invention discloses an N-(-)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1 - Crystalline polymorph A of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, which exhibits a powder x-ray diffraction spectrum comprising from about 50% to about 90%, as shown in Figure 5 The peak identified in the powder x-ray diffraction spectrum. In certain embodiments, the powder x-ray diffraction spectrum is substantially the same as the powder x-ray diffraction pattern shown in FIG.

In a particular embodiment, the invention discloses an N-(-)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1 Crystalline polymorph A of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide exhibiting a differential scanning calorimetry spectrum substantially identical to the differential scanning calorimetry spectrum shown in Figure 6. .

In a particular embodiment, the present invention discloses an N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, a polymorph of 3-dihydroxypropyl)cyclopropane-1-sulfonamide which crystallizes amorphous N-(3,4-difluoro-2-() by a mixture comprising ethyl acetate and heptane. Process for the preparation of 2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. In certain embodiments, the mixture of ethyl acetate and heptane is in a ratio of from about 1 to about 4 parts ethyl acetate to from about 2 to about 10 parts heptane.

In a particular embodiment, the present invention discloses a pharmaceutical composition comprising an effective amount of the crystalline polymorph disclosed herein.

In a particular embodiment, the invention features a method of inhibiting a MEK enzyme comprising: comprising a MEK enzyme comprising: and The compound composition of the compound is contacted. In certain embodiments, the MEK enzyme is MEK kinase.

In a particular embodiment, the invention features a method of treating a MEK-mediated disorder comprising administering to a subject in need thereof a package comprising: A composition of the compound. In certain embodiments, the MEK-mediated condition is a cytokine-mediated condition. In certain embodiments, the MEK-mediated disorder is selected from the group consisting of an immune disorder, an inflammatory disorder, an infectious disorder, a proliferative disorder, or a combination thereof. In certain embodiments, the MEK mediated condition is cancer. In certain embodiments, the MEK-mediated condition is a fibrotic disorder. In certain embodiments, the _{Tmax of} the compound is achieved at 0.5 hours and 5 hours after the composition is administered to a fasted individual. In certain embodiments, the compound achieves a _Cmax between about 0.01 [mu]g/ml to about 1.0 [mu]g/ml on Day 1 when administered to an individual. In certain embodiments, the compound has an AUC between about 0.1 μg hour/mL to about 5.0 μg hour/mL from 0 to 12 hours. In certain embodiments, the compound has a plasma concentration of greater than about 0.01 mg/mL after a single dose of about 5 hours.

In a particular embodiment, the invention features a method of inhibiting growth of a plurality of target cells comprising administering to a subject in need thereof comprising: and A composition of the compound. In certain embodiments, the target cells are cancer cells. In certain embodiments, the _{Tmax of} the compound is achieved at 0.5 hours and 5 hours after the composition is administered to a fasted individual. In certain embodiments, the compound achieves a _Cmax between about 0.01 [mu]g/ml to about 1.0 [mu]g/ml on Day 1 when administered to an individual. In certain embodiments, the compound has an AUC between about 0.1 μg hour/mL to about 5.0 μg hour/mL from 0 to 12 hours. In certain embodiments, the compound has a plasma concentration of greater than about 0.01 mg/mL after a single dose of about 5 hours.

The novel features of the invention are set forth with particular reference to the appended claims. A better understanding of the features and advantages of the present invention can be obtained by referring to the following description of the embodiments of the exemplary embodiments and the accompanying drawings.

The section headings used herein are for organizational purposes only and are not intended to be construed as limiting the invention. All documents or portions of documents referred to in this application, including but not limited to, patents, patent applications, papers, books, manuals, and protocol standards are expressly incorporated herein by reference.

I. Specific terms

Unless otherwise defined, 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 subject matter claimed.

It is to be understood that the foregoing description of the invention and the embodiments of the invention are intended to In the present application, the use of the singular includes the plural unless otherwise specified. It must be noted that the singular forms "a", "an", "the" and "the" are meant to include the plural. It should also be understood that the use of "or" means "and/or" unless otherwise specified. In addition, the use of "including" and other forms such as "include", "includes" and "included" is not restricted.

The definitions of standard chemical terms can be found in reference books, including "ADVANCED ORGANIC CHEMISTRY4 ^TH ED." by Carey and Sundberg, Vols. A (2000) and B (2001), Plenum Press, New York. Nomenclature and analytical chemistry, synthetic organic chemistry, and pharmaceutical and pharmaceutical chemistry experimental procedures and techniques used in analytical chemistry, synthetic organic chemistry, and pharmacy and medicinal chemistry described herein are familiar to those skilled in the art unless otherwise specified. Well known.

Unless otherwise indicated, the use of a generic chemical term such as, but not limited to, "alkyl", "amine", "aryl" is equivalent to its mutated form. For example, "alkyl" as used herein includes optionally substituted alkyl.

As used herein, "moiety", "chemical group", "group" and "chemical group" mean a specific fragment or functional group of a molecule. Chemical groups are often recognized as chemical entities embedded in or attached to a molecule.

The term "bond" or "single bond" means a bond between two atoms or two groups, in which case the atom to which the bond is bonded is considered to be part of a larger structure.

The terms "optional" or "optionally" are used to mean that subsequent events or circumstances are or may not occur, and the description includes examples of occurrences or occurrences of the events or circumstances. An example that does not happen. For example, "optionally substituted alkyl" means "alkyl" or "substituted alkyl" as defined below. Further, the optionally substituted group includes unsubstituted (for example, -CH ₂ CH ₃ ), fully substituted (for example, -CF ₂ CF ₃ ), and monosubstituted (for example, -CH ₂ CH). ₂ F) or a degree of substitution anywhere between a complete substitution and a mono-substitution (e.g., -CH ₂ CHF ₂ , -CH ₂ CF ₃ , -CF ₂ CH ₃ , -CFHCHF _{2 ,} etc.). Those skilled in the art will be aware of any group containing one or more substituents which are not intended to introduce any substitution or substitution pattern (for example, substituted alkyl groups include optionally substituted naphthenes). A base, which is in turn defined to include an optionally substituted alkyl group, potentially infinitely large, which is tridimensionally impractical and/or not readily synthesizable. Thus, any of the substituents described should be broadly understood to have a maximum molecular weight of about 1,000 otaves, and more typically up to about 500 amps (except where the macromolecular substituents are clearly propensity, such as multi-peptide, Glycans, polyethylene glycols, DNA, RNA and the like).

Unless otherwise stated, general chemical terms, such as, but not limited to, "alkyl", "amine" and "aryl" are unsubstituted.

As used herein, C ₁ -C _x includes C ₁ -C ₂ , C ₁ -C ₃ ... C ₁ -C _x . By way of example only, a group designated "C ₁ -C ₄ " indicates that there are from 1 to 4 carbon atoms in the group, that is, one carbon atom, two carbon atoms, three carbon atoms or four a group of carbon atoms, and a range of C ₁ -C ₂ and C ₁ -C ₃ . Thus, by way of example only, "C ₁ -C ₄ alkyl" indicates from 1 to 4 carbon atoms in the alkyl group, that is, selected from methyl, ethyl, propyl, isopropyl, n-butyl, An alkyl group of an isobutyl group, a second butyl group, and a third butyl group. Whenever it occurs herein, a numerical range such as "1 to 10" means each integer in a specific range; for example, "1 to 10 carbon atoms" means that the group has 1 carbon atom, 2 Carbon, 3, 4, 5, 6, 5, 7, 8 or 9 carbon atoms.

As used herein, the terms "A and A' together with the carbon atom to which they are attached form a 3- to 6-membered saturated ring" refer to the following structures used in the compounds disclosed herein:

The terms "heteroatom" or "hetero" as used herein, alone or in combination, mean an atom that is not carbon or hydrogen. The heteroatoms are independently selected from the group consisting of oxygen, nitrogen, sulfur, phosphorus, antimony, selenium and tin, but are not limited to such atoms. In embodiments in which two or more heteroatoms are present, two or more heteroatoms are identical to each other. In embodiments in which two or more heteroatoms are present, two or more heteroatoms are different from each other.

The term "alkyl" as used herein, alone or in combination, means a straight or branched saturated hydrocarbon unit having from 1 to about 10 carbon atoms or from 1 to 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, 2,2-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, 2,2-dimethyl 1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, second butyl, tert-butyl, n-pentyl Base, isopentyl, neopentyl, third amyl and hexyl, and longer alkyl such as heptyl, octyl and the like. Whenever it appears in this context, a numerical range such as "C ₁ -C ₆ alkyl" or "C _1-6 alkyl" means 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 An alkyl group consisting of a carbon atom, 5 carbon atoms or 6 carbon atoms. In one embodiment, "alkyl" is substituted. Unless otherwise specified, "alkyl" is unsubstituted.

The term "alkenyl" as used herein, alone or in combination, means a straight or branched hydrocarbon mono-group 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. . In certain embodiments, this group is in a cis configuration near the double bond. In certain embodiments, this group is in a trans configuration near the double bond. Examples include, but are not limited to, vinyl (-CH=CH ₂ ), 1-propenyl (-CH ₂ CH=CH ₂ ), isopropenyl [-C(CH ₃ )=CH ₂ ], butenyl, 1,3-butadienyl and its analogs. Whenever present herein, a numerical range such as "C ₂ -C ₆ alkenyl" or "C _2-6 alkenyl" means that the alkenyl group may be 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, It consists of 5 carbon atoms or 6 carbon atoms. In one embodiment, "alkenyl" is substituted. Unless otherwise specified, "alkenyl" is unsubstituted.

The term "alkynyl" as used herein, alone or in combination, means a straight or branched hydrocarbon mono-group having one or more carbon-carbon bonds and having from 2 to about 10 carbon atoms or from 2 to about 6 carbon atoms. . Examples include, but are not limited to, alkynyl, 2-propynyl, 2-butynyl, 1,3-butadiynyl, and the like. Whenever present herein, a numerical range such as "C ₂ -C ₆ alkynyl" or "C _2-6 alkynyl" means that the alkynyl group may be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, It consists of 5 carbon atoms or 6 carbon atoms. In one embodiment, "alkynyl" is substituted. Unless otherwise specified, "alkynyl" is unsubstituted.

The terms "heteroalkyl", "heteroalkenyl" and "heteroalkynyl" as used herein, alone or in combination, mean, respectively, alkyl, alkenyl and alkynyl structures as defined above, wherein one or more backbone chain carbon atoms And (if appropriate, any associated hydrogen atom) are independently heteroatoms (ie, atoms that are not carbon, such as, but not limited to, oxygen, nitrogen, sulfur, antimony, phosphorus, tin, or combinations thereof), or Heteroatomic substitutions such as, but not limited to, -OO-, -SS-, -OS-, -SO-, =NN=, -N=N-, -N=N-NH-, -P(O) _2- , -OP(O) ₂ -, -P(O) ₂ -O-, -S(O)-, -S(O) ₂ -, -SnH ₂ - and the like.

The terms "haloalkyl", "haloalkenyl" and "haloalkynyl" as used herein, alone or in combination, mean alkyl, alkenyl and alkynyl as defined above, wherein one or more hydrogen atoms are Substituted by fluorine, chlorine, bromine or iodine atoms or a combination thereof. In certain embodiments, two or more hydrogen atoms are substituted with the same halogen atom as each other (eg, difluoromethyl); in other embodiments, two or more hydrogen atoms are different from each other. Substituted by a halogen atom (for example, 1-chloro-1-fluoro-1-iodoethyl). Non-limiting examples of haloalkyl are fluoromethyl, chloromethyl and bromoethyl. A non-limiting example of a haloalkenyl group is a bromovinyl group. A non-limiting example of a haloalkynyl group is a chloroethynyl group.

The term "carbon chain" as used herein, alone or in combination, means any alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl or heteroalkynyl group, such as linear, cyclic or any combination thereof. If the chain is a moiety of a linking group and the linking group comprises one or more rings which are part of the core skeleton, in order to calculate the length of the chain, the "chain" only includes the bottom or top of the specific ring. For both carbon atoms, and when the top and bottom of the ring are not equal, a shorter distance should be used to determine the chain length. If the chain contains a heteroatom as part of the backbone, such atoms are not counted as part of the carbon chain length.

The terms "cycle", "cyclic", "ring" and "number of members" used herein, alone or in combination, mean any covalently closed structure, including the fats as described herein. Ring, heterocyclic, aromatic, heteroaromatic, and multimembered ring fused or non-fused ring systems. The situation was replaced. The ring forms part of a fused ring system. The term "member" means the number of skeleton atoms constituting the ring. Thus, by way of example only, cyclohexane, pyridine, piperazine, and pyrimidine are 6-membered rings and cyclopentane, pyrrole, tetrahydrofuran, and thiophene are 5-membered rings.

The term "fused" as used herein, alone or in combination, means that two or more rings share one or more bonded ring structures.

The term "cycloalkyl" as used herein, alone or in combination, means a saturated hydrocarbon monocyclic ring containing from 3 to about 15 ring carbon atoms or from 3 to about 10 ring carbon atoms, although additional, non-cyclic rings may be included. A carbon atom is used as a substituent (for example, methylcyclopropyl). Whenever it appears herein, a numerical range such as "C ₃ -C ₆ cycloalkyl" or "C _3-6 cycloalkyl" means that the cycloalkyl group may be 3 carbon atoms, 4 carbon atoms, 5 carbons. Atom or 6 carbon atoms, that is, cyclopropyl, cyclobutyl, cyclopentyl or cycloheptyl, although the definition herein also encompasses the occurrence of the term "cycloalkyl" without a specified range of values. This term includes fused, non-fused, bridged, and spiro groups. In certain embodiments, a fused cycloalkyl contains two to four fused rings, wherein the attached ring is a cycloalkyl ring, and the other independent rings are alicyclic, heterocyclic, aromatic, hetero Aromatic 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 of the description include, but are not limited to, the following groups:

And its analogues.

In one embodiment, "cycloalkyl" is substituted. Unless otherwise specified, this "cycloalkyl" group is unsubstituted.

The terms "non-aromatic heterocyclic group" and "heteroalicyclic group" as used herein, alone or in combination, mean a saturated, partially unsaturated or fully unsaturated non-aromatic ring containing from 3 to about 20 ring atoms. A group in which one or more ring atoms are atoms other than carbon, which are independently selected from the group consisting of oxygen, nitrogen, sulfur, phosphorus, antimony, selenium, and tin, but are not limited thereto. In certain embodiments, wherein two or more heteroatoms are present in the ring, the two or more heteroatoms are identical to each other. In certain embodiments, two or more heteroatoms are present in the ring, and some or all of the two or more heteroatoms are different from each other. This term includes fused, non-fused, bridged, and spiro groups. In certain embodiments, the fused non-aromatic heterocyclic group contains two to four fused rings, wherein the attached ring is a non-aromatic heterocyclic ring, and the other independent rings are alicyclic, heterocyclic, aromatic Family, heteroaromatic or any combination thereof. In certain embodiments, the fused ring system is a bridged single bond or double bond fused, and is a cross-linking bond of a carbon-carbon, carbon-heteroatom or heteroatom-hetero atom. The term also includes groups having from 3 to about 12 backbone ring atoms, as well as groups having from 3 to about 10 backbone ring atoms. In certain embodiments, a non-aromatic heterocyclic subunit is attached to its parent molecule via a heteroatom or carbon atom. Also, in certain embodiments, the additional substitution is via a heteroatom or a carbon atom. As a non-limiting example, the imidazolium non-aromatic heterocyclic ring is via its N atom (imidazolidin-1-yl or imidazolidin-3-yl) or any of its carbon atoms (imidazolidin-2-yl, imidazole) A pyridin-4-yl or imidazolidin-5-yl) is attached to the parent molecule. In a particular embodiment, the non-aromatic heterocyclic ring contains one or more carbonyl or thiocarbonyl groups such as pendant-containing and sulfur-containing groups. Examples include, but are not limited to, pyrrolidinyl, tetrahydrofuran, dihydrofuran, tetrahydrothiophenyl, tetrahydropyranyl, dihydropiperidyl, tetrahydrothiopiperidyl, hexahydropyridyl, morpholinyl , thiomorpholinyl, thiooxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, high Hexahydropyridyl, oxepanyl, thiepanyl, oxazepinyl, diazepyl, thiazepinyl, 1 , 2,3,6-tetrahydropyridyl, 2-pyrroline, 3-pyrrolyl, porphyrin, 2H-piperidyl, 4H-piperidyl, dioxanyl, 1 , 3-dioxolanyl, pyrazolinyl, dithiaalkyl, dithiolanyl, dihydropiperidyl, dihydrothienyl, dihydrofuranyl, pyrazolyl, imidazolinyl, Imidazolidinyl, 3-azabicyclo[3.1.0]hexane, 3-azabicyclo[4.1.0]heptyl, 3H-indenyl and quinazinyl. Illustrative examples of heterocycloalkyl are also referred to as non-aromatic heterocycles, which include:

And its analogues.

These terms also include all cyclic forms of hydrocarbons including, but not limited to, monosaccharides, disaccharides, and oligosaccharides. In one embodiment, the "non-aromatic heterocyclic group" or "heteroalicyclic group" is substituted. Unless otherwise specified, the "non-aromatic heterocyclic group" or "heteroalicyclic group" is unsubstituted.

The term "aryl" as used herein, alone or in combination, means an aromatic hydrocarbon containing from 6 to about 20 ring carbon atoms, and includes both fused and non-fused aromatic rings. In certain embodiments, a fused aromatic ring group contains two to four fused rings, wherein the attached ring is an aromatic ring. In certain embodiments, the other independent ring is an aliphatic ring, a heterocyclic ring, an aromatic ring, a heteroaromatic ring, or any combination thereof. Moreover, the term aryl includes both 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; fused ring aryl-containing naphthyl, phenanthryl, anthracenyl, fluorenyl; and non-fused bisaryl includes biphenyl. In one embodiment, "aryl" is substituted. Unless otherwise specified, this "aryl" is unsubstituted.

The term "heteroaryl" as used herein, alone or in combination, means an aromatic mono-group containing from about 5 to about 20 backbone ring atoms, wherein one or more ring atoms are heteroatoms independently selected from oxygen, Nitrogen, sulfur, phosphorus, antimony, selenium and tin are, but not limited to, such atoms, and provided that the ring of the group does not contain two adjacent O or S atoms. In the embodiment in which two or more hetero atoms are present in the ring, two or more hetero atoms are identical to each other, or part or all of two or more hetero atoms are different from each other. The term heteroaryl includes fused and non-fused heteroaryl groups having at least one hetero atom. The term heteroaryl also includes fused and non-fused heteroaryl groups having from 5 to about 12 skeletal ring atoms, and from 5 to about 10 skeletal ring atoms. In certain embodiments, the heteroaryl group is bonded via a carbon atom or a hetero atom. As a non-limiting example, the imidazolyl group is via any of its carbon atoms (imidazol-2-yl, imidazol-4-yl or imidazol-5-yl) or its nitrogen atom (imidazol-1-yl or imidazol-3-yl) ) Bonded to the parent molecule. Also, in certain embodiments, the heteroaryl is further substituted via any or all of its carbon atoms or any or all of its heteroatoms. In certain embodiments, a fused heteroaryl contains two to four fused rings wherein the attached ring is a heteroaryl ring and the other independent ring is a heterocyclic ring, an aromatic ring, a heteroaromatic ring, or any combination thereof. Non-limiting examples of monocyclic heteroaryl groups include pyridyl; fused ring heteroaryl groups include benzimidazolyl, quinolyl, acridinyl; and non-fused biheteroaryl includes bipyridyl. Other examples of heteroaryl groups include, but are not limited to, furyl, thienyl, oxazolyl, acridinyl, cyanoazinyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl , benzothiadiazolyl, benzothienyl, benzooxadiazolyl, benzotriazolyl, imidazolyl, indolyl, isoxazolyl, isoquinolinyl, indolizinyl , isothiazolyl, isodecyloxadiazolyl, oxazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrrolyl, pyrazinyl, pyrazolyl, indolyl, pyridazinyl, Acridinyl, quinolyl, quinazolinyl, quinolyl, triazolyl, tetrazolyl, thiazolyl, triazinyl, thiadiazolyl, and the like, and oxides thereof, such as pyridyl -N-oxide. Illustrative examples of heteroaryl groups include the following groups:

And its analogues.

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

The term "heterocyclyl" as used herein, alone or in combination, refers to a heteroalicyclic group and a heteroaryl group. Herein, whenever a number of carbon atoms in a heterocyclic ring (for example, a C ₁ -C ₆ heterocyclic ring) is specified, at least one non-carbon atom (hetero atom) must be present in the ring. The reference to "C ₁ -C ₆ heterocycle" means only the number of carbon atoms in the ring and not the total number of atoms in the ring. The term "4-6 member heterocyclic ring" as used herein means the total number of atoms contained in the ring (ie, a four-, five-, or six-membered ring in which at least one atom is a carbon atom, at least one atom is a hetero atom and the remainder 2 to 4 atoms are carbon atoms or heteroatoms). For heterocycles having two or more heteroatoms, the two or more heteroatoms are the same or different from each other. The non-aromatic heterocyclic group includes a group having only three atoms in the ring, and the aromatic heterocyclic group must have at least 5 atoms in the ring. In certain embodiments, the linkage (ie, attached to the parent molecule or further substitution) to the heterocycle is via a heteroatom. In certain embodiments, the bond to the heterocyclic ring is via a carbon atom. In one embodiment, the "heterocyclyl" is substituted. Unless otherwise specified, this "heterocyclic group" is unsubstituted.

The terms "halogen", "halo" or "halide" as used herein, alone or in combination, mean fluoro, chloro, bromo and/or iodine.

Used herein, the term "amino", alone or in combination means a monocyclic group -NH _2.

The term "alkylamino" as used herein, alone or in combination, means a mono-NH(alkyl) group, wherein alkyl is as defined herein.

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

The term "diaminoalkyl" as used herein, alone or in combination, means an alkyl group containing two amine groups, wherein the amine groups are substituents on the alkyl group, which are amine groups, alkyl amine groups or a dialkylamino group, or wherein one or both of the amine groups can form part of an alkyl chain to form a -alkyl-N(H or alkyl)-alkylene-N (H or alkane Or alkyl-) (H or alkyl or alkyl-).

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

The term "cyano" as used herein, alone or in combination, means a mono-CN.

The term "cyanomethyl" as used herein, alone or in combination, means a mono-CH ₂ CN.

Used herein, the term "nitro", alone or in combination means a monocyclic group -NO _2.

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

The term "sideoxy" as used herein, alone or in combination, means that the diradical = O.

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

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

The term "alkoxy" as used herein, alone or in combination, means an alkyl ether group, ie, an -O-alkyl group, which includes a group -O-aliphatic group and an -O-carbocyclic group, wherein the alkyl group, The aliphatic and carbocyclic groups are 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, second butoxy, third butoxy and analog.

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

The term "sulfonyl" as used herein, alone or in combination, means bis-S(=O) _2- .

As used herein, the terms "sulfonamide", "sulfamido" and "sulfamidyl", alone or in combination, mean bis-S(=O) _2- NH- and -NH. -S(=O) ₂ -.

As used herein, the terms "sulfonamide", "sulfamido" and "sulfamidyl", alone or in combination, mean bis-NH-S(=O) ₂ -NH-.

The term "reactant" as used herein refers to a nucleophilic or electrophilic agent used to produce a covalent bond.

It will be appreciated that where two or more groups are used continuously to define a substituent attached to a structure, the first named group is considered to be a terminal group and the last named group is considered to be attached to The structure. Thus, for example, a group arylalkyl group is attached to the structure via an alkyl group.

Used herein, the term "MEK inhibitor" is intended to mean presenting to the MEK activity assay The kinase Mek1 generally described herein of the compounds of no greater than about measured IC ₅₀ of about 50μM or no greater than 100μM. "IC ₅₀ " is an inhibitor that reduces the concentration of enzyme activity (eg, MEK) to a half-maximum level. The compounds disclosed herein have also been found to exhibit inhibition of MEK. For compounds of the present invention is preferably presented in Mek1 MEK kinase assay herein the measurement of not greater than about 10μM, more preferably not greater than about 5μM, more preferably not greater than about 1μM, and most preferably not greater than about 200nM of IC _50.

The term "individual" as used herein encompasses both mammals and non-mammals. None of these terms are interpreted as requiring supervision by a medical professional (eg, a physician, nurse, caregiver, shelter worker). Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates such as chimpanzees and other baboon and monkey breeds; farm animals such as cattle, horses, sheep, goats, pigs; livestock animals such as rabbits , dogs and cats; experimental animals include 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.

As used herein, the terms "treat", "treating" or "treatment" and other grammatical equivalents mean that the progression of the condition is slowed or stopped, causing the deterioration of the condition and alleviating the symptoms of the condition. The prevention of the progression of additional symptoms or the presentation of additional symptoms, mitigation and/or prevention of the underlying cause of the symptoms or a combination thereof. This term further includes the advantage of achieving prevention. For prophylactic advantages, the compounds or compositions disclosed herein can be administered to an individual who is at risk of developing a particular condition, tends to develop a particular condition, or is administered an individual who reports one or more physiological symptoms of the condition.

The term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" as used herein means an amount of an agent or compound sufficient to treat a condition. In certain embodiments, the result is a slowing and/or alleviation of the condition, symptom or cause of the condition, or any other expected change in the biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is required to treat a significant medically significant condition. The appropriate "effective" amount in any independent case can be determined using techniques such as dose increment studies.

The terms "substantially anhydrous" and "substantially solvent free" as used herein mean crystalline polymorphs comprising 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 spectrum or a differential scanning calorimetry spectrum that is different from the ones described herein, but may be within experimental error when considered by those skilled in the art. .

The term "pharmaceutically acceptable" as used herein means a material, such as a carrier or diluent, which does not destroy the biological activity or properties of the compounds described herein and is relatively non-toxic (ie, the material is cast Interacting with the individual does not result in improper biological utility or in an unfavorable manner with any of the components included in the composition).

The term "modulate" as used herein, refers to altering target activity, whether directly or indirectly interacting with a target, including, by way of example only, promoting target activity, inhibiting target activity, limiting target activity, or extending targets. active.

II.MEK

In certain cases, Ras is a signal transduction protein. In certain cases, Ras is activated by the binding of guanosine nucleotides, ie GTP (guanosine triphosphate) or GDP (guanosine diphosphate).

In certain instances, activation of Ras results in activation of the cascade of serine/threonine kinases. In certain instances, activated Ras activates the Raf protein. In certain cases, the activated Raf protein activates "MEK1" and "MEK2".

MEK1 and MEK2 are bifunctional serine/threonine and tyrosine protein kinases, which in specific cases activate MAPK. In certain instances, activation of MAP kinase activated by mitogen cleavage material appears to induce cell proliferation. In certain cases, the activation of MAPK induces cell transformation. In certain cases, blockade of downstream Ras signaling, such as by blocking with dominant negative Raf-1 protein, inhibits mitosis, whether by cell surface receptors or by oncogene Ras mutants Inducing inhibition of the action of cleavage substances.

III. Compound

Standard techniques are used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, and individual delivery and treatment. The reaction and purification techniques are carried out, for example, using kits of the manufacturer's instructions or generally available in the art or as described herein.

Throughout the specification, the groups and substituents thereof can be selected by those skilled in the art to provide stable groups and compounds.

The compounds presented herein may have one or more stereocenters and each stereocenter exists in R or S configuration or in a combination thereof. Similarly, the compounds presented herein may have one or more double bonds and each may be in E (trans) or Z (cis) configuration or in a combination thereof. The presentation of a particular stereoisomer, ortho-isomer, non-image, isomer or epi-iso isomer is understood to include all possible stereoisomers, ortho-isomers, Non-image isomers, mirror image isomers or surface isomers and mixtures thereof. Accordingly, the compounds presented herein include all isolated stereoisomers, regioisomeric, non-image, heterogeneous or epimeric forms, and mixtures thereof. Presenting a particular chemical structure or chemical name of a compound containing one or more palmitic centers, but without specifying a particular stereochemistry, it should be understood to include all stereoisomers, including all possible A stereoisomer, a pure form or a substantially pure form of a particular stereoisomer and a mixture of the other stereoisomer in its pure form or in substantially pure form. Techniques for inverting or leaving a particular stereocenter that has not changed and techniques for dissolving a mixture of stereoisomers are known in the art, and those skilled in the art are fully capable of selecting an appropriate method depending on the particular situation. See, for example, VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5. sup. TH ED., Longman Scientific and Technical Ltd., Essex, 1991 , 809-816; and Heller, Acc. Chem. Res . 1990 , by Furniss et al. 23,128.

Substituents are designated by their conventional chemical formula, which is written from left to right, which equally comprises substituents of the same chemical nature resulting from right to left writing. As a non-limiting example, -CH ₂ O- is equivalent to -OCH ₂ -.

A specific embodiment disclosed herein is a compound of formula I:

Wherein Z is H or F; X is F, Cl, CH ₃ , CH ₂ OH, CH ₂ F, CHF ₂ or CF ₃ ; Y is I, Br, Cl, CF ₃ , C ₁ -C ₃ alkyl, C _2- C ₃ alkenyl, C ₂ -C ₃ alkynyl, cyclopropyl, OMe, OEt, SMe, phenyl or Het, wherein Het is a 5- to 10-membered monocyclic or bicyclic heterocyclic group, the group a group of saturated, olefinic or aromatic groups containing from 1 to 5 ring heteroatoms independently selected from N, O and S; wherein all such phenyl or Het groups are optionally F, Cl, Br, I, B Mercapto, methyl, CN, NO ₂ , CO ₂ H, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, C ₁ -C ₃ alkyl-C(=O)-, C ₁ - C ₃ alkyl-C(=S)-, C ₁ -C ₃ alkoxy-C(=S)-, C ₁ -C ₃ alkyl-C(=O)O-, C ₁ -C ₃ alkane -O-(C=O)-, C ₁ -C ₃ alkyl-C(=O)NH-, C ₁ -C ₃ alkyl-C(=NH)NH-, C ₁ -C ₃ alkyl -NH-(C=O)-, di-C ₁ -C ₃ alkyl-N-(C=O)-, C ₁ -C ₃ alkyl-C(=O)N(C ₁ -C ₃ alkane ,), C ₁ -C ₃ alkyl-S(=O) ₂ NH- or trifluoromethyl substituted; all such methyl, ethyl, C ₁ -C ₃ alkyl and cyclopropyl groups may be optionally OH substitution; all such methyl groups may be substituted by one, two or three F atoms; R ⁰ is H, F, Cl, Br, I, CH ₃ NH-, (CH ₃ ) ₂ N-, C ₁ -C ₆ alkyl, C ₁ -C ₄ alkoxy, C ₃ -C ₆ cycloalkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, phenyl, monosubstituted phenyl, O(C ₁ -C ₄ alkyl), OC(=O)(C ₁ -C ₄ alkyl) or C(=O)O(C ₁ -C ₄ Alkyl); wherein the alkyl, alkoxy, cycloalkyl, alkenyl, alkynyl and phenyl groups are optionally independently selected from the group consisting of F, Cl, Br, I, OH, CN, cyanide Substituted with a substituent of a nitro group, a phenyl group and a trifluoromethyl group; the C ₁ -C ₆ alkyl group and the C ₁ -C ₄ alkoxy group may also be optionally substituted with OCH ₃ or OCH ₂ CH ₃ ; G ₁ , G ₂ , R _1a , R _1b , R _1c , R _1d , R _1e , Ar ₁ , Ar ₂ or Ar ₃ ; wherein G ₁ is optionally an amine group, a C ₁ -C ₃ alkylamino group Or a dialkylamino group substituted C ₁ -C ₆ alkyl group, the dialkylamino group comprising two identical or different C ₁ -C ₄ alkyl groups; or G ₁ is a C ₃ -C ₈ diaminoalkane G ₂ is a 5- or 6-membered ring which is a saturated, unsaturated or aromatic ring containing from 1 to 3 ring heteroatoms independently selected from N, O and S, optionally 1-3 independent Selected from F, Cl, OH, O(C ₁ -C ₃ alkyl), OCH ₂ , OCH ₂ CH ₃ , CH ₃ C(=O)NH, CH ₃ C (=O)O, CN, CF ₃ and a substituent of a 5-membered aromatic heterocyclic group containing 1-4 ring heteroatoms independently selected from N, O and S; R _1a is a methyl group, as the case may be Substituted by 1-3 fluorine atoms or 1-3 chlorine atoms, or substituted by OH, cyclopropoxy or C ₁ -C ₃ alkoxy, wherein the cyclopropoxy or the C ₁ -C ₃ alkoxy The C ₁ -C ₃ alkyl group may be optionally substituted by a hydroxyl group or a methoxy group, and wherein all of the C ₃ alkyl groups in the C ₁ -C ₄ alkoxy group may be further substituted with a second OH group; R _1b It is _{CH (CH 3) -C 1.} 3 alkyl or C ₃ -C ₆ cycloalkyl, such alkyl and cycloalkyl groups optionally 1-3 substituents independently selected from F, Cl, Br, I, OH Substituting a substituent of OCH ₃ and CN; R _1c is (CH ₂ ) _n O _m R'; 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 C ₁ -C ₆ alkyl, which may optionally be selected from 1-3 independently selected from F, Cl, OH, OCH ₃ , OCH ₂ CH ₃ and C ₃ -C ₆ cycloalkyl Substituted by a substituent; R _1d is C(A)(A')(B)-; wherein B is H or C _1-4 alkyl, which may be optionally substituted with one or two OH groups; A and A' independent It is H or C _1-4 alkyl, which is optionally substituted with one or two OH Substituted; or A and A 'together with the carbon atom form a 3- to 6-membered saturated ring; R _1e is

Wherein q is 1 or 2; and R ₂ and R ₃ are each independently H, F, Cl, Br, CH ₃ , CH ₂ F, CHF ₂ , CF ₃ OCH ₃ , OCH ₂ F, OCHF ₂ , OCF ₃ , ethyl , n-propyl, isopropyl, cyclopropyl, isobutyl, t-butyl, t-butyl or methanesulfonyl; R ₄ is H, F, Cl, Br, CH ₃ , CH ₂ F, CHF ₂ , CF ₃ OCH ₃ , OCH ₂ F, OCHF ₂ , OCF ₃ , ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, t-butyl, tert-butyl, methylsulfonium Base, nitro, acetamidine, sulfhydryl, cyano, amine carbaryl, methylamine carbaryl, dimethylamine carbhydryl, 1,3,4-oxadiazol-2-yl, 5-methyl-1,3,4-oxadiazole, 1,3,4-thiadiazole, 5-methyl-1,3,4-thiadiazole 1H-tetrazolyl, N-morpholinyl a carbonylamino group, an N-morpholinylsulfonyl group and an N-pyrrolidylcarbonylamino group; R ₅ is H, F, Cl or methyl; R ₆ is H, F, Cl or methyl; Ar ₁ is

Wherein U and V are independently N, CR ₂ or CR ₃ ; R ₂ , R ₃ and R _{4 are} independently H, F, Cl, Br, CH ₃ , CH ₂ F, CHF ₂ , CF ₃ , OCH ₃ , OCH ₂ F, OCHF ₂ , OCF ₃ , ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, t-butyl, tert-butyl, acetoguanyl, fluorenyl, cyano, amine Sulfhydryl, methylamine, mercapto, dimethylamine, mercapto, 1,3,4-oxadiazol-2-yl, 5-methyl-1,3,4-oxadiazole, 1,3 , 4-thiadiazole, 5-methyl-1,3,4-thiadiazolyl, 1H-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrole a pyridylcarbonylamino group and a methylsulfonyl group; R ₅ and R _{6 are} independently H, F, Cl or methyl; Ar ₂ is

Wherein the dotted line indicates the selective formal position of the double bond in the second ring; 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=; R ₇ is H or methyl; R ₃ is H, acetamino group, A Base, F or Cl; Ar ₃ is

Wherein U is -NH-, -NCH ₃ - or -O-; and R ₇ and R _{3 are} independently H, F, Cl or methyl.

In addition to the definitions given to groups G, R ⁰ , X, Y and Z herein, it also includes additional substitutions that would be expected by those skilled in the art of chemistry and medical technology.

In certain embodiments, the invention provides a compound of Formula I, wherein G is G ₁ or G ₂ . In other embodiments, G is G ₁ . In certain embodiments, G is G ₂ .

In certain embodiments, the invention provides a compound of Formula I, wherein G is R _1a , R _1b , R _1c , R _1d , R _1e , Ar ₁ , Ar ₂ or Ar ₃ . In certain embodiments, G is R _1a , R _1b , R _1c , R _1d or R _1e . In certain embodiments, G is R _1a . In certain embodiments, G is R _1b . In certain embodiments, G is R _1c . In certain embodiments, G is R _1d . In certain embodiments, G is R _1e . In certain embodiments, G is Ar ₁ , Ar ₂ or Ar ₃ . In certain embodiments, G is Ar ₁ . In certain embodiments, G is Ar ₂ . In certain embodiments, G is Ar ₃

In certain embodiments, the compounds disclosed herein are provided as pharmaceutically acceptable salts, solvates, polymorphs, esters, guanamines, tautomers, prodrugs, or combinations thereof.

In certain embodiments, Z is H. In certain embodiments, Z is F. In certain embodiments, X is F. In certain embodiments, X is Cl. In certain embodiments, X is CH _3. In certain embodiments, X is CH ₂ OH. In certain embodiments, X is CH ₂ F. In certain embodiments, X is CHF ₂ . In certain embodiments, X is CF ₃ . In certain embodiments, X is F, Cl or CH _3.

In certain embodiments, G is G ₁ or G ₂ , X is F, Cl or CH ₃ ; Y is I, Br, Cl, CF ₃ , C ₁ -C ₃ alkyl, phenyl, pyridyl, pyrrole The base, pyrazolyl, the phenyl, pyridyl, pyrrolyl and pyrazolyl may be optionally subjected to F, Cl, Br, I, acetyl, methyl, CN, NO ₂ , CO ₂ H, C ₁ - C ₃ alkyl, C ₁ -C ₃ alkoxy, C ₁ -C ₃ alkyl-C(=O)-, C ₁ -C ₃ alkyl-C(=S)-, C ₁ -C ₃ alkane Oxy-C(=S)-, C ₁ -C ₃ alkyl-C(=O)O-, C ₁ -C ₃ alkyl-O-(C=O)-, C ₁ -C ₃ alkyl -C(=O)NH-, C ₁ -C ₃ alkyl-C(=NH)NH-, C ₁ -C ₃ alkyl-NH-(C=O)-, di-C ₁ -C ₃ alkane -N-(C=O)-, C ₁ -C ₃ alkyl-C(=O)N(C ₁ -C ₃ alkyl)-, C ₁ -C ₃ alkyl-S(=O) ₂ NH- or trifluoromethyl substituted; and Z is H or F. In certain embodiments, G is G ₁ or G ₂ and R ⁰ is F, Cl, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy, the C ₁ -C ₄ alkyl group and C ₁ -C ₄ alkoxy groups C ₁ -C ₄ alkyl optionally substituted with F, Cl, OCH ₃ or OCH ₂ CH ₃ group. In certain embodiments, G is G ₁ or G ₂ and R ⁰ is H, F, Cl, C ₁ -C ₄ alkyl, methoxy, ethoxy, or 2-methoxy-ethoxy base.

In certain embodiments, G ₁ is N-methyl-2-aminoethyl. In certain embodiments, G ₁ is (CH ₃ ) ₂ N-CH ₂ CH ₂ —NH—(CH ₂ ) n —, wherein n is 1, 2, or 3. In certain embodiments, G ₁ is (CH ₃ ) ₂ N-CH ₂ CH ₂ —NH—(CH ₂ ) n —, wherein n is 1, 2, or 3, and X is F. In certain embodiments, G ₁ is (CH ₃ ) ₂ N-CH ₂ CH ₂ —NH—(CH ₂ ) _n —, wherein n is 1, 2, or 3, X is F, and Z is F.

In certain embodiments, G ₂ is 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, or 4-piperidinyl. In certain embodiments, G ₂ is morpholinyl, 1-piperazinyl or 2-piperazinyl.

In certain embodiments, G is R _1a , R _1b , R _1c , R _1d , R _1e , Ar ₁ , Ar _{2 ,} or Ar ₃ and X is F, Cl, or CH ₃ . In certain embodiments, G is R _1a , R _1b , R _1c , R _1d , R _1e , Ar ₁ , Ar _{2 ,} or Ar ₃ , X is F, Cl, or CH ₃ and Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl. In certain embodiments, G is R _1a , R _1b , R _1c , R _1d , R _1e , Ar ₁ , Ar _{2 ,} or Ar ₃ , X is F, Cl, or CH ₃ , and Y is I, Br, Cl, CF ₃ , or C ₁ -C ₃ alkyl and Z is H or F.

In certain embodiments, G is _{R 1a, R 1b, R 1c} , R 1d, R 1e, Ar 1, A r2 , or Ar ₃ and R ⁰ is F, Cl, C ₁ -C ₄ -alkyl or C ₁ -C ₄ alkoxy, the C ₁ -C ₄ alkyl and the C ₁ -C ₄ alkoxy groups C ₁ -C ₄ alkyl optionally substituted with F, Cl, oCH ₃ or OCH ₂ CH ₃ group. In certain embodiments, G is R _1a , R _1b , R _1c , R _1d , R _1e , Ar ₁ , Ar _{2 ,} or Ar ₃ and R ⁰ is H, F, Cl, C ₁ -C ₄ alkyl, Methoxy, ethoxy or 2-methoxy-ethoxy.

In certain embodiments, G is R _1a ; and Z is F. In certain embodiments, G is R _1a , wherein R _1a is CH ₃ , R ⁰ is H; and Y is Br, I, CF ₃ or CH ₃ . In certain embodiments, G is R _1b and Z is F. In certain embodiments, G is R _1b , Z is F, and R ⁰ is H, F, or OCH ₃ . In certain embodiments, G is R _1b , Z is F, R ⁰ is H, F or OCH ₃ , and X is F or CH ₃ . In certain embodiments, G is R _1b , Z is F, R ⁰ is H, F or OCH ₃ , X is F or CH ₃ and Y is Br, I or CH ₃ . In certain embodiments, G is R _1b , wherein R _1b is C ₃ -C ₆ cycloalkyl. In certain embodiments, G is R _1b , wherein R _1b is substituted C ₃ -C ₆ cycloalkyl. In certain embodiments, G is R _1b , wherein R _1b is unsubstituted C ₃ -C ₆ cycloalkyl. In certain embodiments, G is R _1b , wherein R _1b is unsubstituted C ₃ -C ₆ cycloalkyl and R ⁰ is H. In certain embodiments, G is R _1b , wherein R _1b is isopropyl or cyclopropyl.

In certain embodiments, G is R _1c and Y is I, Br, CH ₃ or CF ₃ . In certain embodiments, G is R _1c , Y is I, Br, CH ₃ or CF ₃ , and Z is F. In certain embodiments, G is R _1c , Y is I, Br, CH ₃ or CF ₃ , Z is F and m is 0.

In certain embodiments, G is R _1d and R ⁰ is fluoro, chloro, methyl, ethyl, propyl, isopropyl, t-butyl, isobutyl, tert-butyl, cyclopropyl, Cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino. In certain embodiments, G is R _1d and R ⁰ is fluoro, chloro, methyl, ethyl, propyl, isopropyl, t-butyl, isobutyl, tert-butyl, cyclopropyl, Cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino and X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoro methyl. In certain embodiments, G is R _1d and R ⁰ is fluoro, chloro, methyl, ethyl, propyl, isopropyl, t-butyl, isobutyl, tert-butyl, cyclopropyl, Cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino, X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoro Methyl, and Y is I, Br, Cl, or monofluoromethyl, difluoromethyl or trifluoromethyl. In certain embodiments, G is R _1d and R ⁰ is fluoro, chloro, methyl, ethyl, propyl, isopropyl, t-butyl, isobutyl, tert-butyl, cyclopropyl, Cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethylamino, X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoro Methyl, Y is I, Br, Cl, or monofluoromethyl, difluoromethyl or trifluoromethyl and Z is H or F. In certain embodiments, G is R _1d and R ⁰ is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy.

In certain embodiments, G is R _1d , R ⁰ is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy and X is F, Cl or CH ₃ . In certain embodiments, G is R _1d , R ⁰ is F, Cl, methyl, ethyl, methoxy, ethoxy, or 2-methoxy-ethoxy, and X is F, Cl, or CH ₃ and Y are I, Br, Cl, or monofluoromethyl, difluoromethyl or trifluoromethyl. In certain embodiments, G is R _1d , and R ⁰ is F, Cl, methyl, ethyl, methoxy, ethoxy or 2-methoxy-ethoxy, and X is F, Cl or CH. ₃ , Y is I, Br, Cl, or monofluoromethyl, difluoromethyl or trifluoromethyl and Z is H or F. In certain embodiments, G is R _1d and R ⁰ is H; X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoromethyl. In certain embodiments, G is R _1d , R ⁰ is H; X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoromethyl and Y is I, Br, Cl, Or monofluoromethyl, difluoromethyl or trifluoromethyl. In certain embodiments, G is R _1d , R ⁰ is H; X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoromethyl and Y is I, Br, Cl, Or monofluoromethyl, difluoromethyl or trifluoromethyl and Z is H or F.

In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is C ₁ -C ₆ cycloalkyl. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is C ₁ -C ₆ cycloalkyl and B is H. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is methyl, ethyl , 2-hydroxyethyl, n-propyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 3,4-dihydroxybutyl, isopropyl, 1-methyl-2-hydroxyethyl, N-butyl, t-butyl, isobutyl, or 2-hydroxymethyl-3-hydroxypropyl.

In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-di Hydroxypropyl or 3,4-dihydroxybutyl. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-di Hydroxypropyl or 3,4-dihydroxybutyl, wherein the palmitic carbon in B is in the R configuration. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-di Hydroxypropyl or 3,4-dihydroxybutyl, wherein the palmitic carbon of B is the S configuration. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is C ₁ -C ₆ cycloalkyl and B is methyl, which is visible The case is substituted with an OH group, or a C ₂ -C ₄ alkyl group, which may optionally be substituted with one or two OH groups. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and R ⁰ is fluoro, chloro, Methyl, ethyl, propyl, isopropyl, t-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methyl Amino or dimethylamino. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is C ₁ -C ₆ cycloalkyl and R ⁰ is F, Cl, Methyl, ethyl, methoxy, ethoxy, or 2-methoxy-ethoxy. In certain embodiments, G is R _1d, wherein R _1d is C (A) (A ') and C (A) (A') is a C ₁ -C ₆ cycloalkyl and R ⁰ is H; X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoromethyl.

In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-dihydroxypropyl or 3,4-dihydroxybutyl, wherein the palmitic carbon of B is an R configuration which is substantially free of the S isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-dihydroxypropyl, wherein the palmitic carbon of B is an R configuration which is substantially free of the S isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 3,4-dihydroxybutyl, wherein the palmitic carbon of B is an R configuration which is substantially free of the S isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-dihydroxypropyl or 3,4-dihydroxybutyl, wherein the palmitic carbon of B is the S configuration, which is substantially free of the R isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 2,3-dihydroxypropyl, wherein the palmitic carbon of B is the S configuration, which is substantially free of the R isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is C ₁ -C ₆ cycloalkyl and B is 3,4-dihydroxybutyl, wherein the palmitic carbon of B is the S configuration, which is substantially free of the R isomer.

In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is cyclopropyl. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is cyclopropyl and B is H. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is cyclopropyl and B is methyl, ethyl, 2-hydroxyethyl Base, n-propyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 3,4-dihydroxybutyl, isopropyl, 1-methyl-2-hydroxyethyl, n-butyl, Dibutyl, isobutyl or 2-hydroxymethyl-3-hydroxypropyl. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropyl and B is 2,3-dihydroxypropyl or 3 , 4-dihydroxybutyl. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropyl and B is 2,3-dihydroxypropyl or 3 , 4-dihydroxybutyl, wherein the palmitic carbon in B is the R configuration. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropyl and B is 2,3-dihydroxypropyl or 3 , 4-dihydroxybutyl, wherein the palmitic carbon in B is the S configuration. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropyl and B is methyl, optionally via an OH group Substituted, or C ₂ -C ₄ alkyl, which is optionally substituted with one or two OH groups. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropyl and R ⁰ is fluoro, chloro, methyl, ethyl , propyl, isopropyl, t-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, fluoromethyl, methoxy, fluoromethoxy, methylamino or dimethyl Amino group. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropyl and R ⁰ is F, Cl, methyl, ethyl , methoxy, ethoxy, or 2-methoxy-ethoxy. In certain embodiments, G is R _1d , wherein R _1d is C(A)(A′) and C(A)(A′) is cyclopropyl and R ⁰ is H; X is F, Cl, CH ₃ , or monofluoromethyl, difluoromethyl or trifluoromethyl.

In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropane And B is 2,3-dihydroxypropyl or 3,4-dihydroxybutyl, wherein the palmitic carbon in B is an R configuration which is substantially free of the S isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropane And B is a 2,3-dihydroxypropyl group, wherein the palmitic carbon in B is an R configuration which is substantially free of the S isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropane Base and B are 3,4-dihydroxybutyl, wherein the palmitic carbon in B is an R configuration which is substantially free of the S isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropane And B is 2,3-dihydroxypropyl or 3,4-dihydroxybutyl, wherein the palmitic carbon in B is an S configuration which is substantially free of the R isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula 1, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropane And B is a 2,3-dihydroxypropyl group, wherein the palmitic carbon of B is an S configuration, which is substantially free of the R isomer. In certain embodiments, the present invention provides a composition comprising a compound of Formula I, wherein G is R _1d , wherein R _1d is C(A)(A') and C(A)(A') is cyclopropane Base and B are 3,4-dihydroxybutyl, wherein the palmitic carbon in B is an S configuration which is substantially free of the R isomer.

In certain embodiments, G is R _1e and n is 1. In certain embodiments, G is R _1e , R ⁰ is H, R _4-6 is H, and R ₂ and R _{3 are} independently H, F, Cl, Br, CH ₃ , CH ₂ F, CHF ₂ , CF ₃ , OCH ₃ , OCH ₂ F, OCHF ₂ , OCF ₃ , ethyl, n-propyl, isopropyl, cyclopropyl, isobutyl, t-butyl, tert-butyl, and methylsulfonyl, X is F and Y is I.

In certain embodiments, G is Ar ₁ , wherein Ar ₁ is optionally selected from the group consisting of an acetamino group, a decyl group, a cyano group, an amine carbaryl group, a methylamine carbhydryl group, and a dimethylamine group. Indenyl, 1,3,4-oxadiazol-2-yl, 5-methyl-1,3,4-oxazolyl, 1,3,4-thiadiazolyl, 5-methyl-1, Group substitution of 3,4-thiadiazolyl, 1H-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrolidylcarbonylamino and methylsulfonyl The phenyl group is optionally substituted with 1-3 substituents selected from the group consisting of F, Cl and CH ₃ . In certain embodiments, G is Ar ₁ , wherein Ar ₁ is optionally selected from the group consisting of an acetamino group, a decyl group, a cyano group, an amine carbaryl group, a methylamine carbhydryl group, and a dimethylamine group. Indenyl, 1,3,4-oxadiazol-2-yl, 5-methyl-1,3,4-oxadiazolyl, 1,3,4-thiadiazolyl, 5-methyl-1 a group of 3,4-thiadiazolyl, 1H-tetrazolyl, N-morpholinylcarbonylamino, N-morpholinylsulfonyl, N-pyrrolidylcarbonylamino and methylsulfonyl Substituted phenyl, optionally substituted with 1-3 substituents selected from F, Cl and CH ₃ , R ⁰ is H, X is F, Cl or methyl and Y is Br, I, CF ₃ , C ₁ -C ₃ alkyl, C ₂ -C ₃ alkenyl, C ₂ -C ₃ alkynyl, cyclopropyl, OCH ₃ , OCH ₂ CH ₃ or SCH ₃ . In certain embodiments, G is Ar ₁ , wherein Ar ₁ is And wherein R ₂ and R _{3 are} independently H, F, Cl, CH ₃ , CF ₃ , OCH ₃ . In certain embodiments, G is Ar ₁ , wherein Ar ₁ is And wherein R ₂ and R _{3 are} independently H, F, Cl, CH ₃ , CF ₃ , OCH ₃ , X is F or CH ₃ , Y is I, Br or Cl; and Z is F. In certain embodiments, G is Ar ₁ , wherein Ar ₁ is phenyl or monosubstituted phenyl. In certain embodiments, G is Ar ₁ , wherein Ar ₁ is phenyl or monosubstituted phenyl, X is F or CH ₃ , Y is I, Br or Cl, Z is F; and R ⁰ is F, Methyl, ethyl, methoxy or 2-methoxy-ethoxy. In certain embodiments, G is Ar ₁ , wherein U is N or CR ₂ and V is N. In certain embodiments, G is Ar ₁ , wherein U is N or CR ₂ and V is CR. In certain embodiments, G is Ar ₁ , wherein U is N or CR ₂ , V is CR, R ⁰ is H, X is F, Cl or methyl and Y is Br, I, CF ₃ , C ₁ - C ₃ alkyl, C ₂ -C ₃ alkenyl, C ₂ -C ₃ alkynyl, cyclopropyl, OCH ₃ , OCH ₂ CH ₃ or SCH ₂ .

In certain embodiments, G is Ar ₂ , wherein Ar ₂ is Wherein R ₇ is H or methyl and R ₈ is H, acetamido, methyl, F or Cl. In certain embodiments, G is Ar ₂ , wherein Ar ₂ is Wherein R ₇ is H or methyl, R ₈ is H, ethenyl, methyl, F or Cl, R ⁰ is H, X is F, Cl or methyl, and Y is Br, I, CF ₃ , C ₁ -C ₃ alkyl, C ₂ -C ₃ alkenyl, C ₂ -C ₃ alkynyl, cyclopropyl, OCH ₃ , OCH ₂ CH ₃ or SCH ₃ , and Z is F. In certain embodiments, G is Ar ₂ , wherein Ar ₂ is Wherein U is S or O, V is CH=, and R ₈ is H or CH ₃ , R ₇ is H or methyl, R ₈ is H, ethenyl, methyl, F or Cl, and R ⁰ is H, X is F, Cl or methyl, Y is Br, I, CF ₃ , C ₁ -C ₃ alkyl, C ₂ -C ₃ alkenyl, C ₂ -C ₃ alkynyl, cyclopropyl, OCH ₃ , OCH ₂ CH ₃ or SCH ₃ and Z is F. In certain embodiments, R ⁰ is H. In certain embodiments, R ⁰ is H, X is F or Cl, and Y is Br, I, CH ₂ CH ₃ or SCH ₃

In certain embodiments, G is Ar ₃ , wherein U is —O—.

In certain embodiments, G is R _1a , wherein R _{1a is} as defined above. In certain embodiments, G is R _1a and R ⁰ is H, wherein R _{1a is} as defined above. In certain embodiments, G is R _1a and R ^{0 is} as defined above, is not H, and R _{1a is} as defined above. In certain embodiments, G is R _1a , wherein R _1a is methyl, monohalomethyl, C ₁ -C ₃ alkoxymethyl, or cyclopropoxymethyl. In certain embodiments, G is R _1a , wherein R _1a is methyl, monohalomethyl, C ₁ -C ₃ alkoxymethyl or cyclopropoxymethyl and wherein R ⁰ is F, Cl C ₁ -C ₃ alkyl, monochloro C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, trifluoromethoxy, or 2-methoxy-ethoxy.

In certain embodiments, G is R _1b , wherein R _{1b is} as defined above. In certain embodiments, G is R _1b and R ⁰ is H, wherein R _{1b is} as defined above. In certain embodiments, G is R _1b , R ⁰ is H, and Z is F, wherein R _{1b is} as defined above. In certain embodiments, G is R _1b and R ^{0 is} as defined above, not H, and R _{1b is} as defined above. In certain embodiments, G is R _1b , wherein R _1b is isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, etc., all of which may be the case Substituted by 1 or 2 substituents independently selected from the group consisting of F, Cl, OH and OCH ₃ ; Y is Br, I, methyl or trifluoromethyl. In certain embodiments, G is R _1b , wherein R _1b is isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, which may optionally be 1 or 2 substituents independently selected from F, Cl, OH and OCH ₃ ; Y is Br, I, methyl or trifluoromethyl; and R ⁰ is F, Cl, C ₁ -C ₃ alkyl, Monochloro C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In certain embodiments, G is R _1b , wherein R _1b is isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, etc., all of which may be the case Substituted with 1 Cl or with 1 or 2 OH groups; and Y is Br, I, methyl or trifluoromethyl. In certain embodiments, G is R _1b , wherein R _1b is isopropyl, 2-butyl, 2-pentyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, etc., all of which may be the case Substituted with 1 Cl or with 1 or 2 OH groups; Y is Br, I, methyl or trifluoromethyl; and R ⁰ is F, Cl, C ₁ -C ₃ alkyl, monochloro C ₁ - C ₃ alkyl, C ₁ -C ₃ alkoxy, trifluoromethoxy or 2-methoxy-ethoxy.

In certain embodiments, G is R _1c , wherein R _{1c is} as defined above. In certain embodiments, G is R _1c and R ⁰ is H, wherein R _{1c is} as defined above. In certain embodiments, G is R _1c and R ^{0 is} as defined above, is not H, and R _{1c is} as defined above. In certain embodiments, G is R _1c and R ⁰ is H, wherein R _1c is (CH ₂ ) _n O _m R′, wherein m is 0 or 1, and n is 2 or 3 when m is 1. And when m is 0, n is 1 or 2, and R' is C ₁ -C ₆ alkyl, which is optionally selected from 1-3 independently selected from F, Cl, OH, OCH ₃ , OCH ₂ CH ₃ and C. Substituent substitution of _3- C ₆ cycloalkyl. More specific views of another embodiment, m is 0, n is 1 or 2, and R 'is C ₁ -C ₄ alkyl, which is optionally substituted as hereinbefore defined. More specific views of another embodiment, m is 1, n is 2 or 3, and R 'is C ₁ -C ₄ alkyl, which is optionally substituted as hereinbefore defined. In another more specific embodiment, m is 0, n is 1 or 2, and R' is C ₁ -C ₄ alkyl, which is optionally selected from 1-3 independently selected from OH, OCH ₃ , Cl And a group substituted with a cyclopropyl group.

In certain embodiments, G is R _1d , wherein R _{1d is} as defined above. In certain embodiments, G is R _1d and R ⁰ is H, wherein R _{1d is} as defined above. In certain embodiments, G is R _1d and R ^{0 is} as defined above, is not H, and R _{1d is} as defined above. In certain embodiments, G is R _1d and R ⁰ is H, wherein R _1d is C(A)(A')(B)-, wherein B, A, and A' are independently H or C _1-4 An alkyl group, which is optionally substituted with one or two OH groups or a halogen atom, or A and A ^{1 together} with the carbon atom to which they are attached form a 3 to 6 membered saturated ring, which ring may optionally contain one or two independently selected from O. The heteroatoms of N and S are optionally substituted with one or two groups independently selected from the group consisting of methyl, ethyl, fluoro, chloro, bromo and iodo.

In certain embodiments, G is R _1e , wherein R _{1e is} as defined above. In certain embodiments, G is R _1e and R ⁰ is H, wherein R _{1e is} as defined above. In certain embodiments, G is R _1e and R ^{0 is} as defined above, is not H, and R _{1e is} as defined above.

In certain embodiments, G is Ar ₁ , wherein Ar _{1 is} as defined above. In certain embodiments, G is Ar ₁ and R ⁰ is H, wherein Ar _{1 is} as defined above. In certain embodiments, G is Ar ₁ and R ^{0 is} as defined above, not H, and Ar _{1 is} as defined above.

In certain embodiments, G is Ar ₂ , wherein Ar _{2 is} as defined above. In certain embodiments, G is Ar ₂ and R ⁰ is H, wherein Ar _{2 is} as defined above. In certain embodiments, G is Ar ₂ and R ^{0 is} as defined above, is not H, and Ar _{2 is} as defined above.

In certain embodiments, X is F, Cl, or CH ₃ ; Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, and Z is H or F. In certain embodiments, X is F, Cl or CH ₃ , Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, Z is H or F, and R ⁰ is halogen, C ₁ - C ₆ alkyl, monohalogenated C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, phenyl, monosubstituted phenyl, OR ₃ , OC(=O)R ₄ or C(=O)OR ₅ . In certain embodiments, X is F, Cl, or CH ₃ :Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, Z is H or F, and R ⁰ is furanyl, thienyl , thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl. In certain embodiments, X is F, Cl, or CH ₃ , Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, Z is H or F, and R ⁰ is F, Cl, C _1- C ₄ alkyl, C ₁ -C ₃ alkoxy, trifluoromethoxy or 2-methoxy-ethoxy.

In another more specific embodiment, R _1d is cycloalkyl or 1-alkyl-cycloalkyl, wherein the 1-alkyl group optionally has one or two OH groups or one or two halogen atoms Replace.

In another more specific embodiment, R ⁰ is halogen, C ₁ -C ₆ alkyl, monohalogenated C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₂ -C ₆ olefin a C ₂ -C ₆ alkynyl group, a phenyl group, a monosubstituted phenyl group, OR ₃ , OC(=O)R ₄ or C(=O)OR ₅ ; and R _1d is a cycloalkyl group or a 1-alkyl group a cycloalkyl group, wherein the 1-alkyl group is optionally substituted with one or two OH groups or with one or two halogen atoms.

In another more specific embodiment, R ⁰ is furyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl; and R _1d is cycloalkyl Or a 1-alkyl-cycloalkyl group, wherein the 1-alkyl group is optionally substituted with one or two OH groups or with one or two halogen atoms.

In another more specific embodiment, R _1d is 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 embodiment, R _1d is 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 embodiment, R _1d is cycloalkyl or (1-alkyl)-cycloalkyl, wherein the 1-alkyl group is optionally substituted with one or two OH groups, and wherein R ⁰ ' is F, Cl, C ₁ -C ₃ alkyl, monochloro C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, trifluoromethoxy, or 2-methoxy-ethoxy. In another more specific embodiment, R _1d is tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, each optionally substituted as described above, and Wherein Y is Br, I, methyl or trifluoromethyl. In another more specific embodiment, R _1d is oxazolidinyl, thiazolidinyl, isoxazolidinyl, isothiazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, Piperazinyl or morpholinyl, each optionally substituted as previously described, and wherein Y is Br, I, methyl or trifluoromethyl. In another more specific embodiment, R _1d is cyclopropyl or 1-alkyl-cyclopropyl, wherein the 1-alkyl group is optionally substituted with one or two OH groups, and wherein R ⁰ 'is F, Cl, methyl, ethyl, chloromethyl, C ₁ -C ₂ alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In a more specific embodiment, R _1d is 1-(monohydroxyalkyl)cycloalkyl. In another more specific embodiment, R _1d is 1-(monohydroxyalkyl)cycloalkyl, and wherein R ⁰ 'is F, Cl, methyl, ethyl, chloromethyl, C ₁ -C ₂ Alkoxy, trifluoromethoxy or 2-methoxy-ethoxy. In a more specific embodiment, R _1d is 1-(dihydroxyalkyl)cycloalkyl. In another more specific embodiment, R _1d is 1-(dihydroxyalkyl)cycloalkyl, wherein R ⁰ 'is F, Cl, methyl, ethyl, chloromethyl, C ₁ -C ₂ alkane Oxy, trifluoromethoxy or 2-methoxy-ethoxy.

In another more specific embodiment, U is CR ₂ and V is N. In another more specific embodiment, both U and V are N. In another more specific embodiment, U is CR ₂ and V is CR ₃ .

In still another more specific embodiment, the invention provides a compound of formula I, wherein G is Ar ₁ and Ar ₁ is phenyl or monosubstituted phenyl, and R ⁰ is F, methyl, ethyl, C ₁ -C ₃ alkoxy, trifluoromethoxy or 2-methoxy-ethoxy; X is F, Cl or CH ₃ ; Y is I; and Z is F. In another embodiment, the invention provides a compound of formula I, wherein G is Ar ₁ , wherein Ar ₁ is phenyl or monosubstituted phenyl, R ⁰ is halogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, all such alkyl, cycloalkyl, alkenyl and alkynyl groups, optionally 1-3 selected from halogen, OH Substituted with a substituent of CN, cyanomethyl, nitro, phenyl and trifluoromethyl; or R ⁰ is phenyl, OR ₃ , furyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, Isoxazolyl, pyrrolyl or pyrazolyl. In a more specific embodiment, the invention provides a compound of formula I, wherein A is Ar ₁ , wherein Ar ₁ is phenyl or monosubstituted phenyl, and R ⁰ is F, Cl, C ₁ -C ₃ Alkyl, C ₁ -C ₃ alkoxy, 2-methoxyethoxy, C ₂ -C ₃ alkenyl, C ₂ -C ₃ alkynyl, trifluoromethyl, phenyl, furanyl or thienyl , thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl; X is F, Cl or methyl; Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkane Base; and Z is F.

In another more specific embodiment, the invention provides a compound of formula I, wherein G is Ar ₁ , wherein Ar ₁ is phenyl or monosubstituted phenyl, R ⁰ is H; X is F, Cl or CH ₃ ; Y is Br or I; and Z is F.

In another embodiment, the invention provides a compound of formula I, wherein G is Ar ₂ , wherein Ar ₂ is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl Or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylamine, mercaptoamine, etidinyl, methyl, ethyl, trifluoromethyl or halogen. In another more specific embodiment, the invention provides a compound of formula I, wherein G is Ar ₂ , wherein Ar ₂ is 2-thienyl, 2-furyl, 3-thienyl, 3-furanyl, 2 - pyrrolyl or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylamine, mercaptoamine, ethyl hydrazino, methyl, ethyl, trifluoromethyl or halogen; R ⁰ Not H; X is F, Cl or CH ₃ : Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, and Z is H or F. In another embodiment, the invention provides a compound of formula I, wherein G is Ar ₂ , wherein Ar ₂ is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl Or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylamine, mercaptoamine, ethyl hydrazino, methyl, ethyl, trifluoromethyl or halogen; R ⁰ is F, Cl, C ₁ -C ₃ alkyl, monochloro C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, trifluoromethoxy, methoxy-methoxy or 2-methoxy-B Alkyl; X is F, Cl or CH ₃ ; Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl; and Z is H or F. In another embodiment, the invention provides a compound of formula I, wherein G is Ar ₂ , wherein Ar ₂ is 2-thienyl, 2-furyl, 3-thienyl, 3-furyl, 2-pyrrolyl Or 3-pyrrolyl, all optionally substituted by methoxycarbonyl, methylamine, mercaptoamine, ethyl hydrazino, methyl, ethyl, trifluoromethyl or halogen; R ⁰ is H; X is F, Cl or CH ₃ , Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, and Z is H or F. In another embodiment, the invention provides a compound of formula I, wherein G is Ar ₂ , wherein Ar ₂ is thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl or pyrazolyl, all Alternately substituted with methoxycarbonyl, methylamine, mercaptoamine, ethyl hydrazino, methyl, ethyl, trifluoromethyl or halogen; R ⁰ is H or methoxy; X is F , Cl or CH ₃ , Y is I, Br, Cl, CF ₃ or C ₁ -C ₃ alkyl, and Z is H or F.

A specific embodiment disclosed herein is a compound of Compound A:

In certain embodiments, the compound of Compound A is a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer or prodrug.

A particular embodiment disclosed herein is a compound of Compound B:

In certain embodiments, the compound of Compound B is a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer or prodrug.

A specific embodiment disclosed herein is a compound of Compound A:

The 2-OH carbon is in the R configuration. In certain embodiments, the compound of Compound A is a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer or prodrug. In certain embodiments, the composition is substantially free of the S-isomer of the compound. In certain embodiments, the compound contains less than 10% of the S-isomer of the compound. In certain embodiments, the compound contains less than 5% of the S-isomer of the compound. In certain embodiments, the compound contains less than 1% of the S-isomer of the compound.

A particular embodiment disclosed herein is a compound of Compound B:

The 2-OH carbon is in the R configuration. In certain embodiments, the compound of Compound B is a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer or prodrug. In certain embodiments, the composition is substantially free of the S-isomer of the compound. In certain embodiments, the compound contains less than 10% of the S-isomer of the compound. In certain embodiments, the compound contains less than 5% of the S-isomer of the compound. In certain embodiments, the compound contains less than 1% of the S-isomer of the compound.

A specific embodiment disclosed herein is a compound of Compound A:

The 2-OH carbon is in the S configuration. In certain embodiments, the compound of Compound A is a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer or prodrug. In certain embodiments, the composition is substantially free of the R-isomer of the compound. In certain embodiments, the compound contains less than 10% of the R-isomer of the compound. In certain embodiments, the compound contains less than 5% of the R-isomer of the compound. In certain embodiments, the compound contains less than 1% of the R-isomer of the compound.

A particular embodiment disclosed herein is a compound of Compound B:

The 2-OH carbon is in the S configuration. In certain embodiments, the compound of Compound B is a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer or prodrug. In certain embodiments, the composition is substantially free of the R-isomer of the compound. In certain embodiments, the compound contains less than 10% of the R-isomer of the compound. In certain embodiments, the compound contains less than 5% of the R-isomer of the compound. In certain embodiments, the compound contains less than 1% of the R-isomer of the compound.

In certain embodiments, the compounds disclosed herein achieve a Cmax between about 0.01 [mu]g/ml to about 1.0 [mu]g/ml on day 1. In certain embodiments, the compounds disclosed herein achieve a Cmax between about 0.01 [mu]g/ml to about 0.8 [mu]g/ml on day 1. In certain embodiments, the compounds disclosed herein achieve a Cmax between about 0.03 [mu]g/ml to about 0.5 [mu]g/ml on day 1.

In certain embodiments, the compounds disclosed herein have an AUC of from about 0.1 μg hour/mL to about 5.0 μg hour/mL from 0 to 12 hours. In certain embodiments, the compounds disclosed herein have an AUC of from about 0.1 μg hour/mL to about 4.0 μg hour/mL. In certain embodiments, the compounds disclosed herein have an AUC of from about 0.5 μg hour/mL to about 3.0 μg hour/mL.

In certain embodiments, the compounds disclosed herein have a mean AUC of between about 0.1 [mu]g hour/mL to about 5.0 [mu]g hour/mL. In certain embodiments, the compounds disclosed herein have an AUC between about 0.1 [mu]g hour/mL to about 4.0 [mu]g hour/mL. In certain embodiments, the compounds disclosed herein have an average AUC of between about 0.5 [mu]g hour/mL to about 3.0 [mu]g hour/mL.

In certain embodiments, the _Tmax of a compound disclosed herein is achieved from 1 hour to 3 hours after administration of the composition to a fasted individual. In certain embodiments, the compounds disclosed herein have a _Tmax between 0.5 hours and 5 hours. In certain embodiments, the compounds disclosed herein have a _Tmax between 1.0 hours and 3.0 hours. In certain embodiments, the compounds disclosed herein have a _Tmax between 1.0 hours and 2.5 hours.

In certain embodiments, the compounds disclosed herein have an average _{Tmax of} between 0.5 hours and 5 hours. In certain embodiments, the compounds disclosed herein have an average _Tmax between 1.0 hours and 3.0 hours. In certain embodiments, the compounds disclosed herein have an average _{Tmax of} between 1.0 hours and 2.5 hours.

In certain embodiments, the compounds disclosed herein have a plasma concentration of greater than about 0.01 mg/mL after 5 hours of a single dose. In certain embodiments, the compounds disclosed herein have a plasma concentration of greater than about 0.01 mg/mL after a single dose of 10 hours. In certain embodiments, the compounds disclosed herein have a plasma concentration of greater than about 0.01 mg/mL after 15 hours of a single dose.

Synthesis program

Further disclosed herein are methods of synthesizing the compounds disclosed herein. In certain embodiments, the compounds disclosed herein are made by the methods described below. The following procedures and examples are intended to illustrate such methods. Neither a program nor an instance should be construed as limiting the invention in any way. The compounds disclosed herein can also be synthesized using standard synthetic techniques known to those skilled in the art or by methods known in the art. In addition, the solvents, temperatures, and other reaction conditions presented herein may vary depending on the practice and common knowledge of those skilled in the art.

In certain embodiments, starting materials for the synthesis of the compounds described herein are available from commercial sources such as Aldrich Chemical Company (Wisconsin, USA), Sigma Chemical Company (St. Louis, Missouri, USA). In certain embodiments, the starting material used to synthesize the compounds described herein is a synthetic material. In certain embodiments, the compounds described herein and other related compounds having different substituents are as described in ADVANCED ORGANIC CHEMISTRY 4 ^th Ed., March (Wiley 1992); ADVANCED ORGANIC CHEMISTRY 4 by Carey and Sundberg ^Th Ed., Vols. A and B (Plenum 2000, 2001), and Green and Wuts PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3 ^rd Ed., (Wiley 1 999) (which are incorporated by reference in their entirety by reference. The synthesis of techniques and materials in this article in Chinese). The general methods for preparing the compounds disclosed herein are obtained by reactions known in the art, and in certain embodiments, the reactions are modified by the use of appropriate reagents and conditions to introduce the chemical formulas found herein. Different groups in the middle.

Forming a covalent bond by reaction of an electrophile with a nucleophile

The compounds disclosed herein are modified with different electrophiles or nucleophiles to form functional groups or substituents. The following table is entitled "Examples of Covalent Bonds and Their Precursors" listing examples of selected covalent bonds and their precursor functional groups. The precursor functional groups are shown as electrophilic and nucleophilic groups.

Example 4 of the covalent bond Use of protecting groups

In certain embodiments, it may be desirable to protect the desired reactive functional groups in the final product, such as hydroxyl, amine, imido, thio or carboxy groups to avoid undesired participating reactions. The protecting group is used to block some or all of the reactive groups and prevent such groups from participating in the chemical reaction until the protecting group is removed. In some embodiments, each protecting group can be removed in a different manner. The protecting groups that split under completely different reaction conditions meet the needs of different removals. In certain embodiments, the protecting group is removed by acid, base, and hydrogenolysis. In certain embodiments, groups such as trityl, dimethoxytrityl, acetal, and tert-butyldimethylalkyl are acid labile and amine groups protected by a Cbz group. Used in the presence of protecting carboxyl and hydroxyl reactive groups which can be removed by hydrogenolysis and base labile Fmoc groups. In certain embodiments, the carboxyl and hydroxyl reactive groups are, for example, but not limited to, methyl, in the presence of an amine that is unstablely blocked with an acid such as tributyl methacrylate or an acid urethane. The base of the ethyl and ethenyl group is blocked by an unstable group, and both the acid and the base are stable but can be removed by hydrolysis.

In certain embodiments, the carboxylic acid and hydroxyl reactive groups are also blocked by a protecting group such as a benzyl hydrolyzable removable group, and the amine group bonded to the acid hydrogen bond is unstable with a base such as Fmoc. Broken. In certain embodiments, the carboxylic acid reactive group is protected by conversion to a monoester compound as exemplified herein, or it can be blocked by an oxidatively removable protecting group such as 2,4-dimethoxybenzyl. At the same time, the co-existing amine groups are blocked by the fluoride-labile decyl carbamate.

In certain embodiments, an allyl blocking group is available in the presence of an acid protecting group and a base protecting group. For example, the allyl-blocked carboxylic acid is deprotected by a Pd-catalyzed reaction in the presence of an acid labile carboxylic acid carboxylic acid tert-butyl ester or a base labile acetate amine protecting group. Another form of protecting group is a resin to which a compound or an intermediate can be attached. As long as the residual group is attached to the resin, this functional group is blocked and cannot be reacted. This functional group can be used for the reaction once the resin is released.

The protecting group or blocking group is selected from:

A detailed description of other protecting groups, and techniques that can be used to generate protecting groups and their removal, can be found in Protective Groups in Organic Synthesis by Greene and Wuts, 3rd edition, published by John Wiley & Sons, New York, NY, 1999. And Protective Groups by Kocienski, Thieme Verlag, New York, NY, USA, published in 1994, the contents of which are incorporated herein by reference.

Manufacture of the compounds disclosed herein

The compounds disclosed herein can be made by a variety of methods. The following procedures are intended to illustrate such methods, and the examples presented are intended to illustrate the scope of the invention. Such methods or such examples are not to be construed as limiting the invention in any way.

I. Preparation of the compound of structure VI is as follows

Scheme I above illustrates a method for making a sulfonamide derivative of structure VI . In certain embodiments, the 1,2-diamine derivative (Structure IV ) is prepared in two steps from the desired nitro derivative (Structure I ). In certain embodiments, a compound of Structure IV is reacted with a sulfonium chloride derivative (Structure V , with reference to the next scheme) to form the desired sulfonamide. In certain embodiments, the 1,2-diamine derivative IV is protected for the production of imidazolidone (structure VII ) prior to reaction with the corresponding sulfonium chloride. In certain embodiments, deprotection of 1,2-diamine VIII under basic conditions provides the desired material VI.

II. General Paths for the Synthesis of Compounds of General Structure V are summarized below:

The above Scheme II shows an example of preparing a composite sulfonium chloride. In certain embodiments, structure XX is synthesized from IX , alkylated and converted to the potassium salt XII . In certain embodiments, the salt is treated with SOCl ₂ or POCl ₃ to provide the desired compound. Other more specific procedures for the preparation of unique sulfonium chloride derivatives are reported in the experimental paragraphs.

III. General Path for the Synthesis of Compounds of General Structure XIII is outlined in Scheme 3:

Scheme III above illustrates the preparation of a sulfonamide derivative of the general structure XIII . In certain embodiments, such compounds are structurally VI- linked by using a palladium catalyst under Suzuki conditions. Obtained by acid reaction.

IV. General Path for the Synthesis of Compounds of General Structure XIII is outlined in Scheme 4:

The above Scheme IV illustrates the preparation of a sulfonamide derivative of the general structure XV . In certain embodiments, the vinylsulfonamide (XIV) is reacted with an amine to form a derivative of the general structure XV .

Other forms of compounds disclosed herein Isomers of the compounds disclosed herein

In certain embodiments, the compounds disclosed herein exist as geometric isomers. In certain embodiments, the compounds disclosed herein have one or more double bonds. The compounds provided herein include all cis, trans, omnidirectional, reverse, E (entgegen) and Z (zusammen) isomers and their corresponding mixtures. In certain embodiments, the compounds disclosed herein exist as tautomeric forms. The compounds disclosed herein include all possible tautomers in the formulas described herein. In certain embodiments, the compounds disclosed herein have one or more palm centers and each center can exist in an R or S configuration. The compounds disclosed herein include all non-image, isomeric, and epimeric forms and corresponding mixtures thereof. In additional embodiments of the compounds and methods provided herein, mixtures of mirror image isomers and/or non-image areomers derived from a single preparation step, combination or tautomerism can also be used in the applications described herein. In certain embodiments, the compounds disclosed herein can be formed by reacting a racemic mixture of the compound with an optically active resolving agent to form a pair of non-image isomers, isolating such non-image isomers, and recovering optical purity. The mirror isomers were prepared as individual stereoisomers. In certain embodiments, resolution of the mirror image isomers is carried out using covalent non-image-isomerized derivatives of the compounds described herein, or using dissociable complexes (eg, crystalline non-imagewise salts). In certain embodiments, the non-image isomers may be separated by palm chromatography or separation/resolution techniques depending on solubility. In certain embodiments, the optically pure isomers are then recovered with any resolving agent by any means that does not actually cause racemization. A more detailed description of techniques that can be used to resolve stereoisomers of a compound from its racemic mixture can be found in Jean Jacques, Andre Collet, Samuel H. Wilen, "Enantiomers, Racemates and Resolutions", John Wiley And Sons, Inc., 1981, the disclosures of which are incorporated herein by reference.

Labeled compounds disclosed herein

Labeled compounds are also described herein. In addition, disclosed herein are methods of treating a condition by administering a compound disclosed herein that has been labeled,

In certain embodiments, the compounds disclosed herein are isotopically labeled (ie, comprise a radioactive isotope). In certain embodiments, the compounds disclosed herein are labeled by the use of chromophores or fluorescent groups, bioluminescent labels, or chemiluminescent labels.

Isotopically labeled compounds disclosed herein are also equivalent to those described herein, but wherein one or more of the atoms are replaced by an atomic mass or mass number different from the atomic mass or mass number normally found in nature. In certain embodiments, the isotopes incorporated in the compounds disclosed herein are selected from the group consisting of ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ 0, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. In certain embodiments, the isotopically labeled compounds disclosed herein are used in drug and/or matrix tissue distribution assays. In certain embodiments, the isotopically labeled compounds disclosed herein have greater metabolic stability (eg, increased in vivo half-life or reduced dosage requirements).

In certain embodiments, the compounds disclosed herein can be labeled by any convenient means.

Pharmaceutically acceptable salts of the compounds disclosed herein

Also described herein are pharmaceutically acceptable salts. Also disclosed herein are methods of treating a condition by administering a pharmaceutically acceptable salt of a compound disclosed herein.

In certain embodiments, when the acid proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion; or when coordinating with an organic base, the compound disclosed herein is prepared as a pharmaceutically acceptable Accept the salt.

In certain embodiments, a base addition salt is prepared by reacting a free acid form of a compound disclosed herein with a pharmaceutically acceptable inorganic or organic base, including but not limited to an organic base. Such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methyl reduced glucosamine and the like and inorganic bases such as aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide and the like Things.

In certain embodiments, the compounds disclosed herein are prepared as pharmaceutically acceptable salts by reacting the free base form of the compound with a pharmaceutically acceptable inorganic or organic acid, 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, caproic acid, cyclopentanepropionic acid, glycolic acid, and coke Acid, lactic acid, malonic acid, succinic acid, hydroxysuccinic acid, maleic acid, fumaric acid, Q-toluenesulfonic acid, tartaric acid, trifluoroacetic acid, citric acid, benzoic acid, 3-( 4-hydroxybenzimidyl)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, glucoheptanoic acid, 4,4'-methylenebis-(3-hydroxy- 2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and Diene acid.

Solvates of the compounds disclosed herein

Solvates are also disclosed herein. Also disclosed herein are methods of treating a condition by administering a solvate of a compound disclosed herein.

The solvate contains a stoichiometric or non-stoichiometric amount of solvent. In certain embodiments, solvates of the compounds disclosed herein are formed in a process for crystallization in a pharmaceutically acceptable solvent such as water, ethanol, and the like. In certain embodiments, a hydrate of a compound disclosed herein is formed when the solvent is water. In certain embodiments, the alcoholate is formed when the solvent is an alcohol.

In certain embodiments, solvates of the compounds disclosed herein are prepared or formed in the processes described herein. In certain embodiments, hydrates of the compounds disclosed herein are prepared by recrystallization from an aqueous/organic solvent mixture using an organic solvent including, but not limited to, dioxane, tetrahydrofuran or methanol. In certain embodiments, the compounds disclosed herein are not solvated. Generally, the solvated forms are considered equivalent to the unsolvated forms for use in the compounds and methods provided herein.

Polymorphic forms of the compounds disclosed herein

Polymorphs are also disclosed herein. Also disclosed herein are methods of treating disorders by administering a polymorph of the compounds disclosed herein.

As used herein, "polymorph" includes the arrangement of different crystal layers of the same elemental composition of the compound. In certain instances, each polymorph of a compound has a different x-ray diffraction spectrum, infrared spectrum, melting point, density, hardness, crystal form, optical and electrical properties, stability, and solubility. A variety of factors such as recrystallization solvent, crystallization rate and storage temperature may cause the single crystal shape to dominate.

In a particular embodiment, N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-() is disclosed herein. Crystalline polymorph A of 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide: .

In certain embodiments, N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) Crystalline polymorph A of 3-, 3-dihydroxypropyl)cyclopropane-1-sulfonamide exhibits a specific powder x-ray diffraction pattern. In certain embodiments, the powder x-ray diffraction spectrum contains at least about 50% of the peaks shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum contains at least about 70% of the peaks shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum contains at least about 90% of the peaks shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum is substantially the same as the powder x-ray diffraction pattern shown in FIG.

In certain embodiments, N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) Crystalline polymorph A of 3-, 3-dihydroxypropyl)cyclopropane-1-sulfonamide exhibits a specific differential scanning calorimetry spectrum. In certain embodiments, this particular differential scanning calorimetry spectrum is substantially identical to the differential scanning calorimetry spectrum shown in FIG. In certain embodiments, crystalline polymorph A has an initial melting point of about 143 ° C as measured by differential scanning calorimetry.

In certain embodiments, the N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( Crystalline polymorph A of 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide is obtained by crystallizing amorphous N-(S)-(3,4-difluoro-2-(s) from a solvent. It is prepared by 2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide.

In certain embodiments, the N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( The crystalline polymorph A of 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide is prepared by a mixture of its own alkane and ethyl acetate.

Prodrugs of the compounds disclosed herein

This article also discloses prodrugs. Also disclosed herein are methods of treating a condition by administering a prodrug of a compound disclosed herein.

As used herein, a "prodrug" is a form of a compound that, upon administration to an individual and subsequent absorption, is converted to an active or more active material (i.e., a parent). In certain embodiments, the prodrug is converted to the parent by metabolism.

In certain embodiments, a prodrug of a compound disclosed herein has a chemical group that renders the compound less active and/or modulates the solubility of the compound. In certain embodiments, cleavage of the chemical group results in the conversion of the prodrug to the parent. In certain embodiments, the prodrugs of the compounds disclosed herein are easier to administer than the parent. In certain embodiments, prodrugs of the compounds disclosed herein may be bioavailable by oral administration, but the parent may not. In certain embodiments, the prodrugs of the compounds disclosed herein are superior in solubility to the parent.

Non-limiting examples of prodrugs can be the administration of a compound described herein as an ester ("prodrug") to facilitate delivery through a cell membrane wherein water solubility is not conducive to migration but which is then hydrolyzable to a carboxylic acid, the active entity Water solubility is advantageous once inside the cell. A further example of a prodrug may be a short peptide (polyamino acid) bonded to an acid group, wherein the peptide is metabolized to reveal the reactive group.

For a method of designing prodrugs, see, for example, Fedorak et al . , Am. J. Physio.l . , 269: G210-218 (1995); McLoed et al., Gastroenterol , 106: 405-413 (1994); Hochhaus et al., Biomed. , 6: 283-286 (1992); J. Larsen and H. Bundgaard, Int . J. Pharmaceutics , 37, 87 (1987); J. Larsen et al., Int. J. Pharmaceutics , 47, 103 (1988) Sinkula et al . , J. Pharm. Sci. , 64:181-210 (1975); T. Higuchi and V. Stella, Pro-Drugs as Novel Delivery Systems , ACS Symposium Series, Volume 14; and Edward B. Roche , Bioreversible Carriers in Drug Design , American Pharmaceutical Association and Pergamon Press, 1987; and Saulnier et al. (1994), Bioorganic and Medicinal Chemistry Letters, Vol. 4, p. 1985, all of which are incorporated by reference. In this article.

In certain embodiments, prodrugs of the compounds disclosed herein are by non-derived compounds disclosed herein and a suitable aminoguanidine agent such as, but not limited to, 1,1-decyloxyalkyl chloride It is prepared by reacting an acid ester, p-nitrophenyl carbonate or the like.

In certain embodiments, a prodrug of a compound disclosed herein comprises an amino acid residue covalently linked to a free amine, hydroxy or carboxylic acid group of a compound disclosed herein via a guanamine or ester linkage, or two Or a multi-peptide chain of two or more (ie, two, three or four) amino acid residues. In certain embodiments, the amino acid residue comprises a naturally occurring amino acid, 4-hydroxyproline, hydroxy lysine, demosine, isodemosine, 3-methyl Histamine, n-proline, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine and bismuth thiomethionate.

In certain embodiments, a compound having a free amine group, a guanamine group, a hydroxyl group, or a carboxyl group disclosed herein is converted to a prodrug. In certain embodiments, a compound having a free carboxyl group disclosed herein is converted to a prodrug by derivatizing a free carboxyl group to a guanamine or alkyl ester. In certain embodiments, the compounds having free hydroxyl groups disclosed herein are used by, but not limited to, hemisuccinates, phosphates, dimethylaminoacetates, and phosphonium methoxycarbonyl groups. The group is derived from a free hydroxyl group and converted to a prodrug as outlined in Advanced Drug Delivery Reviews 1996 , l . Also included are hydroxy and amine carbamate prodrugs, as well as hydroxy carbonate prodrugs, sulfonates and sulfates.

In certain embodiments, prodrugs of the compounds disclosed herein are produced by derivatizing a hydroxy group to a (decyloxy)methyl group and a (decyloxy)ethyl ether, wherein the thiol group is an alkyl ester, optionally including (but not limited to) a group substituted with an ether, an amine, and a carboxylic acid functional group, or wherein the thiol group is an amino acid ester as described above. Prodrugs of this type are described in J. Med. Chem. 1996 , 39 , 10, which is incorporated herein by reference. In certain embodiments, the free amine is derivatized as a guanamine, sulfonamide or phosphoniumamine. All such prodrug groups can incorporate groups including, but not limited to, ether, amine, and carboxylic acid functional groups.

In certain instances, the sites on the aromatic ring portion of the compounds disclosed herein are susceptible to a variety of metabolic reactions; thus, in certain embodiments, the compounds disclosed herein comprise appropriate substitutions in the aromatic ring structure. base.

IV. How to use

In certain embodiments, a compound or composition disclosed herein is administered to a subject in need thereof to inhibit the MEK enzyme. In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a condition. In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a MEK-mediated condition. In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a proliferative disorder. In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat an inflammatory condition.

In certain embodiments, a compound or composition disclosed herein is administered to a subject in need thereof to inhibit the MEK enzyme. In certain embodiments, the MEK enzyme is MEK kinase. In certain embodiments, the MEK enzyme is MEK1. In certain embodiments, the MEK enzyme is MEK2.

In certain embodiments, the MEK enzyme is inhibited by at least about 1%. In certain embodiments, the MEK enzyme is inhibited by at least about 2%. In certain embodiments, the MEK enzyme is inhibited by at least about 3%. In certain embodiments, the MEK enzyme is inhibited by at least about 4%. In certain embodiments, the MEK enzyme is inhibited by at least about 5%. In certain embodiments, the MEK enzyme is inhibited by at least about 10%. In certain embodiments, the MEK enzyme is inhibited by at least about 20%. In certain embodiments, the MEK enzyme is inhibited by at least about 25%. In certain embodiments, the MEK enzyme is inhibited by at least about 30%. In certain embodiments, the MEK enzyme is inhibited by at least about 40%. In certain embodiments, the MEK enzyme is inhibited by at least about 50%. In certain embodiments, the MEK enzyme is inhibited by at least about 60%. In certain embodiments, the MEK enzyme is inhibited by at least about 70%. In certain embodiments, the MEK enzyme is inhibited by at least about 75%. In certain embodiments, the MEK enzyme is inhibited by at least about 80%. In certain embodiments, the MEK enzyme is inhibited by at least about 90%. In certain embodiments, the MEK enzyme is substantially completely inhibited.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a MEK-mediated condition. In certain embodiments, the MEK-mediated disorder is selected from the group consisting of an inflammatory disorder, a proliferative disorder, an infection, an immune disorder (eg, an autoimmune disorder, psoriasis, rheumatoid arthritis, bone) Arthritis, dry eye and glaucoma), cardiac conditions (eg stroke, reperfusion injury, ischemia, atherosclerosis, heart failure), neurological disorders, fibrotic disorders, metabolic diseases, chronic pain and/or neuropathic pain .

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat an immunological disorder. In certain embodiments, a compound or composition disclosed herein is administered to a subject in need thereof to treat a cytokine-mediated disorder (ie, a condition caused by overproduction or unregulated production of a pro-inflammatory cytokine, pre-inflammatory cells) Hormones include, for example, TNF, IL-1, IL-6, and IL-8). In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof for the treatment of an autoimmune disorder, an inflammatory disorder, an infectious disorder, prostaglandin peroxidase synthase-2 (COX-2) Symptoms, rheumatoid arthritis, inflammatory bowel disease, inflammatory pain, ulcerative colitis, Crohn's disorder, periodontal disease, underarm and ankle joint disease, multiple sclerosis, diabetes, kidney Spherical nephritis, 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, asthma, allergies, respiratory distress syndrome or acute or chronic pancreatitis and multiple sclerosis.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat an inflammatory condition. In certain embodiments, the inflammatory condition is proliferative glomerulonephritis, chronic inflammatory condition, bursitis, acute rheumatoid arthritis, allergy, bursitis, pulmonary fibrosis, septic arthritis , joint adhesion spondylitis, arthritis, asthma, atherosclerosis, chronic inflammatory disease, chronic obstructive pulmonary disease, diabetes (including diabetic retinopathy), diarrhea, eczema, intestinal arthritis, gastritis, gout , gouty arthritis, inflammatory bowel disease, colonic irritability, ulcerative colitis, juvenile arthritis, neuroarthritis, organ transplant rejection, osteoarthritis, osteoporosis, pancreatitis, proliferative kidney Spherical nephritis, pruritic dermatitis, psoriasis, psoriatic arthritis, pulmonary fibrosis, pulmonary inflammation, septic arthritis, reflux esophagitis, respiratory distress syndrome, acute or chronic pancreatitis, rheumatoid arthritis , scleroderma, cystic fibrosis, systemic lupus erythematosus, tendonitis, ulcerative colitis, vitaligo, spondyloarthropathy or Crohn's disease.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a proliferative disorder. In certain embodiments, the proliferative disorder is the growth of abnormal cells. In certain embodiments, the proliferative disorder is a hemangiomas. In certain embodiments, the proliferative disorder is cancer. In certain embodiments, the cancer is a hematological or non-hematologic blood cancer. In certain embodiments, the cancer is selected from the group consisting of multiple myeloma, leukemia, and lymphoma. In certain embodiments, the cancer is selected from the group consisting of acute and chronic leukemia. In certain embodiments, the cancer is selected from the group consisting of acute lymphocytic leukemia (ALL) and acute non-lymphocytic leukemia (ANLL). In certain embodiments, the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML). In certain embodiments, the cancer is selected from the group consisting of Hodgkin's lymphoma and non-Hodgkin's lymphoma. In certain embodiments, the cancer is multiple myeloma. In certain embodiments, the cancer is low, medium, and advanced. In certain embodiments, the cancer is selected from the group consisting of brain cancer, head and neck cancer, lung cancer, breast cancer, reproductive system cancer, digestive system cancer, pancreatic cancer, and urinary system cancer. In certain embodiments, the cancer is upper gastrointestinal cancer or colorectal cancer. In certain embodiments, the cancer is bladder cancer or renal cell carcinoma. In certain embodiments, the cancer is prostate cancer. In certain embodiments, the cancer is breast cancer such as breast ductal carcinoma, medullary carcinoma, colloidal carcinoma, tubular cancer, and inflammatory breast cancer in mammary duct tissue; ovarian cancer, including epithelial ovarian tumors such as adenocarcinoma in the ovary And adenocarcinoma that migrates from the ovary into the abdominal cavity; uterine cancer; cervical cancer, such as adenocarcinoma in the cervical epithelium, including squamous cell carcinoma and adenocarcinoma; prostate cancer, such as prostate cancer selected from the group consisting of: Cancer or adenocarcinoma that migrates to the bone; pancreatic cancer, such as epithelial carcinoma in the pancreatic duct tissue and adenocarcinoma in the pancreatic duct; bladder cancer, such as transitional cell carcinoma in the bladder, urothelial carcinoma (transitional cell carcinoma) ), tumor, squamous cell carcinoma, adenocarcinoma, and small cell carcinoma in the urothelial cells of the inner lining of the bladder; leukemia, such as acute myeloid leukemia (AML), acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic Myeloid leukemia, hairy cell leukemia, myelodysplasia, and myeloproliferative disorders; bone cancer; lung cancer, such as non-small cell lung cancer (NSCLC), differentiates into squamous cell carcinoma, adenocarcinoma, and large cell undifferentiated carcinoma Small cell lung cancer; skin cancer, such as basal cell carcinoma, melanoma, squamous cell carcinoma, and actinic keratosis, which are skin symptoms that sometimes progress to squamous cell carcinoma; ocular retinoblastoma; skin or Intraocular (ocular) melanoma; primary liver cancer (starting to be cancer in the liver); kidney cancer; thyroid cancer, such as papillary, follicular, medulla, and undifferentiated; AIDS-related lymphoma, such as Diffusion of large B-cell lymphoma, B-cell immunoblastic lymphoma, and small non-cleaved cell lymphoma; Kaposi's Sarcoma; virus-induced cancer, including hepatitis B virus (HBV), hepatitis C virus (HCV) and liver cancer; type 1 human lymphoblastic virus (HTLV-1) and adult T-cell leukemia/lymphoma; and human papillomavirus (HPV) and cervical cancer; central nervous system cancer (CNS), Such as primary brain tumors, including glioma (astrocytoma, degenerative astrocytoma or glioblastoma multiforme), oligodendroglioma, ependymoma, meningioma, Lymphoma, schwannomas, and neural tube blastoma; peripheral nervous system (PNS) , such as acoustic neuroma and malignant peripheral nerve sheath tumor (MPNST), including neurofibromatosis and schwannomas, malignant fibroblastoma, malignant fibrous histiocytoma, malignant meningioma, malignant mesothelioma and malignant mixed Mülle Tumor (malignant mixed Oral and oropharyngeal cancer, such as hypopharyngeal, laryngeal, nasopharyngeal, and oropharyngeal cancer; gastric cancer, such as lymphoma, gastric stromal tumor, and carcinoid tumor; testicular cancer, such as blastoma (GCTs), It includes seminoma and non-seminoma and gonadal stromal tumors, including Leydig cell tumor and Sertoli cell tumor; thymic cancer, such as thymoma, Thymic cancer, Hodgkin's disease, non-Hodgkin's lymphoid carcinoid or carcinoid tumor; rectal cancer; colon cancer, kidney cancer, adrenocortical carcinoma, follicular lymphoma, negative pre-B cells (Pre-B) Leukemia, chronic lymphocytic B-leukemia, adenocarcinoma, angiosarcoma, astrocytoma, acoustic neuroma, degenerative astrocytoma, basal cell carcinoma, blastoglioma, chondrosarcoma, choriocarcinoma, chordoma , craniopharyngioma, cutaneous melanoma, cystadenocarcinoma, endothelial sarcoma, embryonal carcinoma, ependymoma, Ewing's tumor, epithelial cancer, fibrosarcoma, gastric cancer, genitourinary tract cancer, polymorphism Glioblastoma, vascular mother Tumor, hepatocellular carcinoma, liver cancer, Kaposi's sarcoma, large cell carcinoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphatic endothelial sarcoma, medullary thyroid carcinoma, neural tube blastoma, meningococcal tumor, Mesothelioma, myeloma, mucinous sarcoma, neuroblastoma, neurofibrosarcoma, oligodendroglioma, osteogenic sarcoma, epithelial ovarian cancer, papillary carcinoma, papillary adenocarcinoma, parathyroid tumor , pheochromocytoma, pineal tumor, plasmacytoma, retinoblastoma, rhabdomyosarcoma, sebaceous gland cancer, spermatogonia, skin cancer, melanoma, small cell lung cancer, squamous cell carcinoma, sweat gland cancer, Synovial tumor, thyroid cancer, uveal melanoma and Wilm's tumor, oral and laryngeal cancer, respiratory cancer, bone and joint cancer, soft tissue cancer, skin cancer, reproductive system cancer, eye and eyelid cancer , nervous system cancer, lymphatic system cancer and endocrine system cancer. In a particular embodiment, the cancer is selected from the group consisting of: tongue, mouth, pharynx or other oral cancer; esophageal cancer, gastric or small intestine cancer; colon cancer or rectal, anal or anorectal cancer; liver, intrahepatic gallbladder Tube, gallbladder, pancreas or other bile or digestive cancer; throat, bronchial and other respiratory cancer; heart cancer, melanoma, basal cell carcinoma, squamous cell carcinoma, other non-epithelial skin cancer; uterus or cervical cancer Uterine body cancer; ovarian, vulva, vagina, or other female genital cancer; prostate, testicular, penile or other male genital cancer; bladder cancer; kidney cancer; kidney, pelvic or urinary tract cancer or other genitourinary cancer; thyroid cancer or Other endocrine cancers; chronic lymphocytic leukemia; and cutaneous T-cell lymphoma, both granular and mononuclear.

In certain embodiments, the compounds and/or compositions disclosed herein are administered to an individual in need thereof to treat a proliferative disorder. In certain embodiments, the proliferative disorder is a fibrotic disorder. In certain embodiments, the proliferative disorder is selected from the group consisting of angiogenesis-related symptoms or symptoms, angioplasty, atherosclerosis, cardiac hypertrophy, hyperplasia, immune disorders, inflammation, interstitial nephritis or pulmonary fibrosis, keloids Formation, cirrhosis, migraine, pain, polymyositis, hyperplasia after medical procedures, restenosis, rheumatoid arthritis, scleroderma, systemic lupus or angiogenesis.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a proliferative disorder. In certain embodiments, the proliferative disorder is benign skin hyperplasia (eg, psoriasis) or prostate (eg, benign prostatic hypertrophy (BPH)).

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a blood disorder. In certain embodiments, the blood disorder is selected from the group consisting of sickle cell anemia, myelodysplastic disorders (MDS), and myeloproliferative disorders. In certain embodiments, the proliferative disorder is selected from the group consisting of polycythemia vera, myelofibrosis, and essential thrombocythemia.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat an ophthalmic condition. In certain embodiments, the ophthalmic condition is dry eye (including Sjogren's syndrome), macular degeneration, angle closure and open angle glaucoma, retinal ganglion cell degeneration, ocular ischemia, retina Inflammation, optic neuropathy (eg, glaucomatous optic neuropathy or diabetic optic neuropathy), uveitis, ocular inflammation, inflammation associated with ocular tissue trauma, and pain (eg, inflammation or pain after surgery for ocular surgery, such as cataract surgery and Refractive surgery).

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a skin condition. In certain embodiments, the dermatological condition is melanoma, basal cell carcinoma, squamous cell carcinoma, psoriasis, and persistent itching.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a metabolic disorder. In certain embodiments, the metabolic disorder is metabolic syndrome, insulin resistance, and Type I and Type II diabetes.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a condition associated with tissue damage. In certain embodiments, the condition associated with tissue damage is selected from the group consisting of a vascular disorder, migraine, nodular periarteritis, thyroiditis, aplastic anemia, Hodgkin's disease, sclerodoma, rheumatic fever Type I diabetes, neuromuscular junction disorders including myasthenia gravis, white matter disorders including multiple sclerosis, sarcoidosis, nephritis, renal syndrome, Behcet's syndrome, polymyositis, gingivitis, Periodontal disease, allergy, post-traumatic swelling, ischemia including myocardial ischemia, cardiovascular ischemia, ischemic secondary to cardiac arrest, allergic rhinitis, respiratory distress syndrome, endotoxin shock syndrome and atherosclerosis Hardening.

In certain embodiments, a compound or composition disclosed herein is administered to an individual in need thereof to treat a cardiovascular condition. In certain embodiments, the cardiovascular condition is atherosclerosis, cardiac hypertrophy, spontaneous cardiomyopathy, heart failure, angiogenesis-related symptoms or conditions, and hyperplasia induced after medical surgery, including but not limited to Restenosis caused by surgery and angioplasty.

The compounds or compositions disclosed herein are administered to an individual in need thereof to treat a neurological condition associated with tissue damage. In certain embodiments, the neurological condition is Parkinson's disease; Alzheimer's disease; Alzheimer's dementia; central nervous system damage caused by stroke, ischemia, and trauma; epilepsy; neuropathic pain , depression; or bipolar disorder.

In certain embodiments, the compounds and/or compositions disclosed herein can be administered to prevent blast cell implantation.

In certain embodiments, the compounds and/or compositions disclosed herein can be administered to degrade cells, inhibit cell growth, or kill cells. In certain embodiments, the cells are cancer cells. In certain embodiments, the cell is a brain, breast, lung, ovary, pancreas, prostate, kidney, or colorectal cancer cell.

In certain embodiments, the compounds and/or compositions disclosed herein can be administered to inhibit the growth of a plurality of target cells. In certain embodiments, target cell growth can be inhibited by about 1%. In certain embodiments, target cell growth can be inhibited by about 2%. In certain embodiments, target cell growth can be inhibited by about 3%. In certain embodiments, target cell growth can be inhibited by about 4%. In certain embodiments, target cell growth can be inhibited by about 5%. In certain embodiments, target cell growth can be inhibited by about 10%. In certain embodiments, target cell growth can be inhibited by about 20%. In certain embodiments, target cell growth can be inhibited by about 25%. In certain embodiments, target cell growth can be inhibited by about 30%. In certain embodiments, target cell growth can be inhibited by about 40%. In certain embodiments, target cell growth can be inhibited by about 50%. In certain embodiments, target cell growth can be inhibited by about 60%. In certain embodiments, target cell growth can be inhibited by about 70%. In certain embodiments, target cell growth can be inhibited by about 75%. In certain embodiments, target cell growth can be inhibited by about 80%. In certain embodiments, target cell growth can be inhibited by about 90%. In certain embodiments, target cell growth can be inhibited by about 100%. In certain embodiments, the target cell is a cancer cell.

In certain embodiments, the compounds and/or compositions disclosed herein can be administered to degrade a plurality of target cells. In certain embodiments, 1% of the target cells are degraded. In certain embodiments, 2% of the target cells are degraded. In certain embodiments, 3% of the target cells are degraded. In certain embodiments, 4% of the target cells are degraded. In certain embodiments, 5% of the target cells are degraded. In certain embodiments, 10% of the target cells are degraded. In certain embodiments, 20% of the target cells are degraded. In certain embodiments, 25% of the target cells are degraded. In certain embodiments, 30% of the target cells are degraded. In certain embodiments, 40% of the target cells are degraded. In certain embodiments, 50% of the target cells are degraded. In certain embodiments, 60% of the target cells are degraded. In certain embodiments, 70% of the target cells are degraded. In certain embodiments, 75% of the target cells are degraded. In certain embodiments, 80% of the target cells are degraded. In certain embodiments, 90% of the target cells are degraded. In certain embodiments, 100% of the target cells are degraded. In certain embodiments, substantially all of the target cells are degraded. In certain embodiments, the target cell is a cancer cell.

In certain embodiments, the compounds and/or compositions disclosed herein can be administered to kill a plurality of target cells. In certain embodiments, 1% of target cells are killed. In certain embodiments, 2% of the target cells are killed. In certain embodiments, 3% of the target cells are killed. In certain embodiments, 4% of the target cells are killed. In certain embodiments, 5% of the target cells are killed. In certain embodiments, 10% of the target cells are killed. In certain embodiments, 20% of the target cells are killed. In certain embodiments, 25% of the target cells are killed. In certain embodiments, 30% of the target cells are killed. In certain embodiments, 40% of the target cells are killed. In certain embodiments, 50% of the target cells are killed. In certain embodiments, 60% of the target cells are killed. In certain embodiments, 70% of the target cells are killed. In certain embodiments, 75% of the target cells are killed. In certain embodiments, 80% of the target cells are killed. In certain embodiments, 90% of the target cells are killed. In certain embodiments, 100% of the target cells are killed. In certain embodiments, the target cell is a cancer cell.

In certain embodiments, the compounds and/or compositions disclosed herein are administered to an individual in need thereof to reduce tumor size, inhibit tumor growth, reduce metastasis, or prevent metastasis.

In certain embodiments, the tumor size is reduced. In certain embodiments, the tumor size is reduced by at least 1%. In certain embodiments, the tumor size is reduced by at least 2%. In certain embodiments, the tumor size is reduced by at least 3%. In certain embodiments, the tumor size is reduced by at least 4%. In certain embodiments, the tumor size is reduced by at least 5%. In certain embodiments, the tumor size is reduced by at least 10%. In certain embodiments the tumor size is reduced by at least 20%. In certain embodiments, the tumor size is reduced by at least 25%. In certain embodiments, the tumor size is reduced by at least 30%. In certain embodiments, the tumor size is reduced by at least 40%. In certain embodiments, the tumor size is reduced by at least 50%. In certain embodiments, the tumor size is reduced by at least 60%. In certain embodiments, the tumor size is reduced by at least 70%. In certain embodiments, the tumor size is reduced by at least 75%. In certain embodiments, the tumor size is reduced by at least 80%. In certain embodiments, the tumor size is reduced by at least 85%. In certain embodiments, the tumor size is reduced by at least 90%. In certain embodiments, the tumor size is reduced by at least 95%.

In certain embodiments, tumor growth is inhibited. In certain embodiments, tumor growth can be inhibited by at least 1% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 2% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 3% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 4% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 5% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 6% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 10% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 20% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 30% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 40% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 50% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 60% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 70% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 75% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 80% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 90% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 95% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, tumor growth can be inhibited by at least 99% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein.

In certain embodiments, the transfer is inhibited. In certain embodiments, the transfer can inhibit at least 1% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 2% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 3% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 4% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 5% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 6% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 10% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 20% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 30% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 40% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 50% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 60% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 70% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 75% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 80% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 90% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 95% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, the transfer can inhibit at least 99% relative to the growth rate prior to administration of the compounds and/or compositions disclosed herein. In certain embodiments, prevention of metastasis.

In certain embodiments, the compounds or compositions disclosed herein are used for diagnostic purposes and/or as research reagents. In certain embodiments, the compounds and/or compositions disclosed herein are used in differential and/or combinatorial analysis to account for the pattern of expression of genes expressed in cells and tissues.

V. Pharmaceutical composition

A specific embodiment disclosed herein is a pharmaceutical composition comprising Compound A or a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer, prodrug, hydrate or derivative thereof. Things.

In certain embodiments, the composition comprises Compound A:

The 2-OH carbon is in the R configuration and is substantially free of S-isomers. In certain embodiments, the 2-OH carbon on the compound is in the R configuration. In certain embodiments, the composition is substantially free of the S-isomer of the compound. In certain embodiments, the compound contains less than 10% of the S-isomer of the compound. In certain embodiments, the compound contains less than 5% of the S-isomer of the compound. In certain embodiments, the compound contains less than 1% of the S-isomer of the compound. In certain embodiments, the compound is in an R configuration.

In certain embodiments, the composition comprises Compound A:

The 2-OH carbon is in the S configuration and is substantially free of R-isomers. In certain embodiments, the 2-OH carbon on the compound is in the S configuration. In certain embodiments, the composition is substantially free of the R-isomer of the compound. In certain embodiments, the compound contains less than 10% of the R-isomer of the compound. In certain embodiments, the compound contains less than 5% of the R-isomer of the compound. In certain embodiments, the compound contains less than 1% of the R-isomer of the compound. In certain embodiments, the compound is in the S configuration.

A specific embodiment disclosed herein is a pharmaceutical composition comprising Compound B or a pharmaceutically acceptable salt, solvate, polymorph, ester, guanamine, tautomer, prodrug, hydrate or derivative thereof. Things.

In certain embodiments, the composition comprises Compound B:

The 2-OH carbon is in the R configuration and is substantially free of S-isomers. In certain embodiments, the 2-OH carbon on the compound is in the R configuration. In certain embodiments, the composition is substantially free of the S-isomer of the compound. In certain embodiments, the compound contains less than 10% of the S-isomer of the compound. In certain embodiments, the compound contains less than 5% of the S-isomer of the compound. In certain embodiments, the compound contains less than 1% of the S-isomer of the compound. In certain embodiments, the compound is in an R configuration.

In certain embodiments, the composition comprises Compound B:

The 2-OH carbon is in the S configuration and is substantially free of R-isomers. In certain embodiments, the 2-OH carbon on the compound is in the S configuration. In certain embodiments, the composition is substantially free of the R-isomer of the compound. In certain embodiments, the compound contains less than 10% of the R-isomer of the compound. In certain embodiments, the compound contains less than 5% of the R-isomer of the compound. In certain embodiments, the compound contains less than 1% of the R-isomer of the compound. In certain embodiments, the compound is in the S configuration.

A particular embodiment disclosed herein is a pharmaceutical composition comprising a compound disclosed herein. In certain embodiments, the compound is present in an amount from about 1 to about 50 mg. In certain embodiments, the compound is present in an amount from about 1 to 10 mg. In certain embodiments, the compound is present in an amount from about 10 to 20 mg. In certain embodiments, the compound is present in an amount from about 20-40 mg. In certain embodiments, the compound is present in an amount from about 40-50 mg.

A particular embodiment disclosed herein is a pharmaceutical composition comprising at least about 50% of a compound exhibiting a powder x-ray diffraction spectrum comprising at least 50% of the peaks identified by the powder x-ray diffraction pattern shown in Figure 5. . In certain embodiments, the powder x-ray diffraction spectrum comprises at least 70% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 90% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum is substantially the same as the powder x-ray diffraction pattern shown in FIG.

In certain embodiments, the composition comprises at least about 75% of a compound exhibiting a powder x-ray diffraction spectrum comprising at least 50% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 70% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 90% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum is substantially the same as the powder x-ray diffraction pattern shown in FIG.

In certain embodiments, the composition comprises at least about 90% of the compound exhibiting a powder x-ray diffraction spectrum comprising at least 50% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 70% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 90% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum is substantially the same as the powder x-ray diffraction pattern shown in FIG.

In certain embodiments, substantially all of the compounds present in the composition exhibit a powder x-ray diffraction spectrum comprising at least 50% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 70% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum comprises at least 90% of the peaks identified by the powder x-ray diffraction pattern shown in FIG. In certain embodiments, the powder x-ray diffraction spectrum is substantially the same as the powder x-ray diffraction pattern shown in FIG.

In certain embodiments, the crystalline polymorph present in the composition has an initial melting point of about 143 ° C as measured by differential scanning calorimetry. In certain embodiments, the crystalline polymorph is substantially anhydrous. In certain embodiments, the crystalline polymorph is substantially solvent free.

In certain embodiments, the composition contains at least about 50% of the compound, which exhibits a differential scanning calorimetry spectrum substantially identical to the differential scanning calorimetry spectrum shown in FIG. In certain embodiments, the crystalline polymorph has an initial melting point of about 143 ° C as measured by differential scanning calorimetry. In certain embodiments, the crystalline polymorph is substantially anhydrous. In certain embodiments, the crystalline polymorph is substantially solvent free.

In certain embodiments, the composition comprises at least about 75% of the compound exhibiting a differential scanning calorimetry spectrum substantially identical to the differential scanning calorimetry spectrum shown in FIG. In certain embodiments, the crystalline polymorph has an initial melting point of about 143 ° C as measured by differential scanning calorimetry. In certain embodiments, the crystalline polymorph is substantially anhydrous. In certain embodiments, the crystalline polymorph is substantially solvent free.

In certain embodiments, the composition contains at least about 90% of the compound, which exhibits a differential scanning calorimetry spectrum substantially identical to the differential scanning calorimetry spectrum shown in FIG. In certain embodiments, the crystalline polymorph has an initial melting point of about 143 ° C as measured by differential scanning calorimetry. In certain embodiments, the crystalline polymorph is substantially anhydrous. In certain embodiments, the crystalline polymorph is substantially solvent free.

In certain embodiments, substantially all of the compounds present in the composition exhibit a differential scanning calorimetry spectrum substantially identical to the differential scanning calorimetry spectrum shown in FIG. In certain embodiments, the crystalline polymorph has an initial melting point of about 143 ° C as measured by differential scanning calorimetry. In certain embodiments, the crystalline polymorph is substantially anhydrous. In certain embodiments, the crystalline polymorph is substantially solvent free.

In certain embodiments, the pharmaceutical composition comprises: a polymorph of N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, which is composed of N-(3,4-difluoro-2-(2-fluoro-4-iodine) containing crystalline amorphous The procedure for the step of phenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide is prepared. In certain embodiments, the crystallization step comprises crystallizing from a mixture of ethyl acetate and heptane, for example, a mixture of ethyl acetate and heptane is from about 1 to 4 parts ethyl acetate to about 2 to 10 parts heptane. The ratio, or more particularly the ratio of from about 2 parts ethyl acetate to about 5 parts heptane.

In certain embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition further comprises an adjuvant, an excipient and a preservative, an agent for delaying absorption, a filler, a binder, an adsorbent, a buffer, a disintegrant, a solubilizer, and the like. Agents, and other inert ingredients.

In certain embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier. Suitable pharmaceutical carriers include inert diluents or fillers, water, and various organic solvents.

In certain embodiments, the composition includes a filler or diluent. In various embodiments, the filler or diluent is selected from the group consisting of microcrystalline cellulose, deuterated microcrystalline cellulose, lactose, mannitol, compressible sugars, calcium phosphate, calcium sulfate, calcium carbonate, calcium citrate, and starch. In other embodiments, the filler or diluent is microcrystalline cellulose.

In certain embodiments, the composition includes 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, saponite, and Magnesium silicate (veegum). In certain embodiments, the disintegrant is croscarmellose sodium.

In certain embodiments, the composition includes a lubricant. In various embodiments, the lubricant is selected from the group consisting of magnesium stearate, metal stearate, talc, sodium stearyl fumarate, and stearic acid. In certain embodiments, the lubricant is magnesium stearate.

In certain embodiments, the composition includes a humectant or a surfactant. In various embodiments, the humectant or surfactant is selected from the group consisting of sodium lauryl sulfate, glycerin, sorbitan oleate, sorbitan stearate, polyoxyethylated sorbitan laurate , palmitate, stearate, oleate or hexaolate, polyoxyethylene stearyl alcohol and sorbitan monolaurate. In certain embodiments, the humectant or surfactant is sodium lauryl sulfate.

Other excipients such as slip agents, perfumes, and color formers may also be added. Other excipients can be found ^{in: The Handbook of PharmaceuticalExcipients, 5 th} Edition, 2005 and the FDA Inactive Ingedient Database.

In certain embodiments, the composition comprises microcrystalline cellulose. In certain embodiments, the composition comprises croscarmellose sodium. In certain embodiments, the composition comprises sodium lauryl sulfate. In certain embodiments, the composition comprises magnesium stearate.

In certain embodiments, the composition further comprises at least one member selected from the group consisting of microcrystalline cellulose, deuterated microcrystalline cellulose, lactose, compressible sugar, xylitol, sorbitol, mannitol, pregelatinized starch, A filler of maltodextrin, calcium phosphate, calcium carbonate, starch and calcium citrate. In certain embodiments, the composition further comprises at least one member selected from the group consisting of croscarmellose sodium, sodium starch glycolate, crospovidone, methylcellulose, alginic acid, sodium alginate, starch derivatives, a disintegrating agent of sapphire and magnesium aluminum silicate. In certain embodiments, the composition further comprises at least one lubricant selected from the group consisting of magnesium stearate, metal stearate, talc, sodium stearyl fumarate, and stearic acid. In certain embodiments, the composition further comprises at least one selected from the group consisting of sodium lauryl sulfate, glycerin, sorbitan oleate, sorbitan stearate, polyoxyethylated sorbitan lauric acid A humectant or surfactant for esters, palmitates, stearates, oleates or hexanoates, polyoxyethylene stearyl alcohol and sorbitan monolaurate.

In certain embodiments, the pharmaceutical composition comprises: about 1 mg of a compound of the following structure: About 222.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate. In certain embodiments, the pharmaceutical composition comprises: about 10 mg of a compound of the following structure: About 213.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 20 mg of a compound of the following structure: About 203.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 40 mg of a compound of the following structure: About 183.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 0.4% by weight of a compound of the following structure: And about 99.6 wt% of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is about 92.6 wt% of the weight of the composition. In certain 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.

In certain embodiments, the pharmaceutical composition comprises: about 4.2% by weight of a compound of the following structure: And about 95.8 wt% of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is about 88.8 wt% of the composition. In certain 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.

In certain embodiments, the pharmaceutical composition comprises: from about 2% to about 10% by weight of a compound of the following structure: And from about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline vegan. In certain embodiments, the microcrystalline cellulose is from about 85 wt% to about 95 wt% of the composition. In certain embodiments, the composition further comprises: from about 1% to about 6 wt% croscarmellose sodium; from about 0.1% to about 2 wt% sodium lauryl sulfate; and from about 0.25 wt% to about 1.5 wt% % magnesium stearate. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is from about 85 wt% to about 95 wt% of the composition. In certain embodiments, the composition further comprises: from about 1 wt% to about 6 wt% croscarmellose sodium; and from about 0.25 wt% to about 1.5 wt% magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 1 mg of a compound of the following structure: About 222.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 10 mg of a compound of the following structure: About 213.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 20 mg of a compound of the structure: viscous; about 203.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of lauryl sulfate Sodium; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 40 mg of a compound of the following structure: About 183.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

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

In certain embodiments, the pharmaceutical composition comprises: about 4.2% by weight of a compound of the following structure: And about 95.8 wt% of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is about 88.8 wt% of the composition. In certain embodiments, the composition further comprises: about 5% by weight of croscarmellose sodium; about 1% by weight of sodium lauryl sulfate; and about 1% by weight of magnesium stearate.

In certain embodiments, the pharmaceutical compositions comprise: from about 2% to about 10% by weight of a compound of the structure: And from about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is from about 85 wt% to about 95 wt% of the composition. In certain embodiments, the composition further comprises: from about 1 wt% to about 6 wt% croscarmellose sodium; from about 0.1 wt% to about 2 wt% sodium lauryl sulfate; and from about 0.25 wt% to about 1.5 wt% % magnesium stearate. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is from about 85 wt% to about 95 wt% of the composition. In certain embodiments, the composition further comprises: from about 1 wt% to about 6 wt% croscarmellose sodium; and from about 0.25 wt% to about 1.5 wt% magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 1 mg of a compound of the following structure: About 222.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

The invention also relates to a composition comprising: about 10 mg of a compound of the following structure: About 213.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 20 mg of a compound of the following structure: About 203.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: about 40 mg of a compound of the following structure: About 183.2 mg of microcrystalline cellulose; about 12.0 mg of croscarmellose sodium; about 2.4 mg of sodium lauryl sulfate; and about 2.4 mg of magnesium stearate.

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

In certain embodiments, the pharmaceutical composition comprises: about 4.2% by weight of a compound of the following structure: And about 95.8 wt% of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is about 88.8 wt% of the composition. In certain embodiments, the composition further comprises: about 5% by weight of croscarmellose sodium; about 1% by weight of sodium lauryl sulfate; and about 1% by weight of magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises from about 2% to about 10% by weight of a compound of the structure: And from about 98% to about 90% by weight of a pharmaceutically acceptable carrier or vehicle. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is from about 85 wt% to about 95 wt% of the composition. In certain embodiments, the composition further comprises: from about 1 wt% to about 6 wt% of croscarmellose sodium; from about 0.1 wt% to about 2 wt% sodium lauryl sulfate; and about 0.25 wt% about 1.5 wt% % magnesium stearate. In certain embodiments, a pharmaceutically acceptable carrier or vehicle comprises microcrystalline cellulose. In certain embodiments, the microcrystalline cellulose is from about 85 wt% to about 95 wt% of the composition. In certain embodiments, the composition further comprises: from about 1 wt% to about 6 wt% croscarmellose sodium; and about 0.25 wt% about 1.5 wt% magnesium stearate.

In certain embodiments, the pharmaceutical composition comprises: N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) Crystalline polymorph A of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. In certain embodiments, the pharmaceutical composition comprises: N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) Crystalline polymorph A of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, and at least one pharmaceutically acceptable carrier.

Formulation

In certain embodiments, the compositions disclosed herein are formulated for oral administration. In certain embodiments, the compositions disclosed herein are administered in the form of troches, capsules, pills, powders, solutions, suspensions, caps, troches, pellets or beads.

In certain embodiments, the compositions disclosed herein are administered via a lozenge. In certain embodiments, the tablet comprises an inert diluent (eg, calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate); granulation and disintegrant (eg, croscarmellose sodium, cross-linked poly-dimensional) Ketone or sodium starch glycolate); fillers (eg microcrystalline cellulose, deuterated microcrystalline cellulose, pregelatinized starch, lactose, dicalcium phosphate or compressible sugar); binders (eg hypromellose, Povidone, starch, gelatin, polyvinylpyrrolidone or gum arabic); surfactants (eg sodium lauryl sulfate) and/or lubricants and processing aids (eg talc, croscarmellose sodium, corn) Starch or alginic acid, magnesium stearate, stearic acid, colloidal cerium oxide and sodium lauryl sulfate. In certain embodiments, the lozenge further comprises a sweetener, a flavoring agent, a coloring agent, and a preservative.

In certain embodiments, the lozenge comprises citric acid, a disintegrant (eg, starch, alginic acid, and a specific complex citrate) and a binder (eg, sucrose, gelatin, and acacia).

In certain embodiments, the tablet is uncoated or coated. In certain cases, the coating will mask the taste of the composition. In certain cases, the coating can improve disintegration and absorption in the gastrointestinal tract.

In certain embodiments, the lozenges disclosed herein are prepared in any convenient manner. In certain embodiments, the lozenges disclosed herein are prepared in a dry blend. In certain embodiments, the compounds disclosed herein are incorporated into a dosage form by dry blending with excipients followed by compression into a lozenge form. In certain embodiments, the compressed tablet is prepared by compressing the active ingredient in a free-flowing form such as a powder or granules in any suitable machine, optionally with a binder, inert diluent or lubricant, interfacial activity. Or a mixture of dispersing agents.

In certain embodiments, the lozenges disclosed herein are prepared in any convenient manner. In certain embodiments, the lozenges disclosed herein are prepared by wet granulation. In certain embodiments, the compounds disclosed herein are added to a dry vehicle and mixed, then the binder solution is added, or the drug is dissolved and added as a portion of the granulation in solution. In wet granulation techniques, if a surfactant is used, the surfactant is added to the dry vehicle or to the binder solution and incorporated into the solution.

In certain embodiments, the compositions disclosed herein are administered via a capsule. In certain embodiments, the capsule is a hard capsule. In certain embodiments, the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin. In certain embodiments, the capsule is a soft capsule. In certain embodiments, the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oil medium such as peanut oil, liquid paraffin or olive oil.

In certain embodiments, the presently disclosed capsules are prepared in any convenient manner. In certain embodiments, the compounds disclosed herein are dissolved in a material (eg, a molten form of high molecular weight polyethylene glycol) that is filled into a hard gelatin capsule shell, followed by bonding and sealing. In certain embodiments, the compounds disclosed herein are dissolved in a high molecular weight polyethylene glycol. In certain embodiments, the mixture is cooled and then filled into gelatin capsules.

In certain embodiments, the composition is in the form of a capsule or lozenge and has a total weight of from about 50 mg to about 1000 mg. In certain embodiments, the composition is in the form of a capsule or lozenge and has a total weight selected from the group consisting of 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, and 500 mg. In certain embodiments, the composition is in the form of a capsule or lozenge and has a total weight of about 240 mg.

In certain embodiments, the composition is in the form of a capsule or lozenge and the dosage form comprises from about 1 to about 50 mg of a compound disclosed herein having a USP acceptable value for content homogeneity of less than about 15.

In certain embodiments, the compounds disclosed herein are administered as an aqueous suspension. In certain embodiments, the aqueous suspension comprises a sweetener or flavoring, coloring matter or dye and, if desired, an emulsifier or suspending agent, and a diluent such as water, ethanol, propylene glycol, glycerin or combinations thereof. In certain embodiments, the aqueous suspension comprises a suspending agent. In certain embodiments, the aqueous suspension comprises sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth, and acacia. In certain embodiments, the aqueous suspension comprises a dispersing or wetting agent. In certain embodiments, the aqueous suspension comprises a naturally occurring phospholipid, such as lecithin, or a condensation product of an alkylene oxide with a fatty acid, such as polyoxyethylene stearate, or ethylene oxide with a long chain aliphatic alcohol. a condensation product, such as heptaethyloxyethyl cetyl alcohol, or a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol, such as polyoxyethylene sorbitan monooleate, or a ring A condensation product of oxyethane with a partial ester derived from a fatty acid and a hexitol anhydride, such as polyethylene sorbitan monooleate. In certain embodiments, the aqueous suspension comprises a preservative. In certain embodiments, the aqueous suspension comprises ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate. In certain embodiments, the aqueous suspension comprises a sweetener. In certain embodiments, the aqueous suspension comprises sucrose, saccharin or aspartame.

In certain embodiments, the compounds disclosed herein are administered as an oily suspension. In certain embodiments, the oily suspension is formulated by suspending the active ingredient in a vegetable oil (eg, peanut oil, olive oil, sesame oil or coconut oil) or in a mineral oil (eg, liquid paraffin). In certain embodiments, the oily suspension comprises a thickening agent (eg, beeswax, hard paraffin or cetyl alcohol). In certain embodiments, the oily suspension comprises a sweetener as previously described. In certain embodiments, the oily suspension comprises an antioxidant (eg, butylated hydroxyanisole or alpha-tocopherol).

In certain embodiments, the compositions disclosed herein are formulated for parenteral injection (eg, via injection or perfusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, Intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and subcutaneous). In certain embodiments, the compositions disclosed herein are administered as a sterile solution, suspension or emulsion.

In certain embodiments, formulations for parenteral administration comprise an aqueous and non-aqueous (oily) sterile injectable solution of the active compound, which may comprise an antioxidant, a buffer, a bacteriostatic agent, and a formulation An isotonic solute of the recipient's blood; and an aqueous and non-aqueous sterile suspension which may include a suspending agent and a thickening agent. In certain embodiments, formulations for parenteral administration include a suitable stabilizer or an agent that increases the solubility of the compound such that a highly concentrated solution can be prepared.

In certain embodiments, the compounds disclosed herein are administered as an aqueous suspension. In certain embodiments, the aqueous suspension comprises water, Ringer's solution, or isotonic sodium chloride solution.

In certain embodiments, the compounds disclosed herein are administered as an oil microemulsion in water, wherein the active ingredient is dissolved in the oil phase. In certain embodiments, the compounds disclosed herein are dissolved in a fatty oil such as sesame oil, or a synthetic fatty acid ester such as ethyl oleate or triglyceride, or a liposome. In certain embodiments, the compounds disclosed herein are dissolved in a mixture of soybean oil and lecithin. In certain embodiments, the oil solution is introduced into a mixture of water and glycerin and processed to form a microemulsion.

In certain embodiments, the composition formulated for parenteral administration is administered as a single bolus injection. In certain embodiments, the parenteral compositions formulated for parenteral administration to a continuous intravenous delivery device is administered (e.g., the type Deltec CADD-PLUS ^TM 5400 intravenous pump).

In certain embodiments, the formulation for injection is presented, for example, in an ampule or multi-dose container together with the added preservative in unit dosage form. In certain embodiments, the formulation for injection is stored in powder form or in a lyophilized (freeze-dried) state that requires only immediate addition of a sterile liquid carrier such as saline or sterile pyrogen-free water prior to use.

In certain embodiments, the formulations disclosed herein are administered in a reservoir formulation. In certain embodiments, the reservoir formulation is administered via implantation (eg, subcutaneous or intramuscular) or by intramuscular injection.

In certain embodiments, the compositions disclosed herein are formulated for topical administration. As described herein, topical administration means that the administration of the composition results in the compound not entering the bloodstream significantly. In certain embodiments, the compositions disclosed herein can be applied to the epidermis, buccal cavity, ear, eye, and/or nose.

In certain embodiments, the compositions formulated for topical administration are formulated in the form of a gel, liniment, lotion, cream, ointment or paste, solution, suspension, emulsion or powder. In certain embodiments, the compositions disclosed herein are administered in the form of an ointment or cream. In certain embodiments, the compositions disclosed herein are administered in the form of a mouthwash. In certain embodiments, the compositions disclosed herein are administered via inhalation.

In certain embodiments, the composition formulated for administration via inhalation is delivered by an insufflator, nebulizer, pressurized pack, or other device that facilitates aerosol spray delivery. The pressurized pack may contain suitable propellants such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. In the case of a pressurized aerosol, the dosage unit is determined by providing a valve to deliver a metered amount. Alternatively, for inhalation or insufflation, the pharmaceutical preparation may be in the form of a dry powder composition, for example a powder mixture of the compound and a suitable powder base such as lactose or starch. The powder composition is presented in unit dosage form, for example in a capsule, filter cartridge, gelatin or blister pack, for administration of the powder therefrom by means of an inhaler or insufflator. For buccal or sublingual administration, the compositions may take the form of lozenges, buccal tablets, tablets or gels formulated in a conventional manner. The compositions may comprise the active ingredient in a flavoring base such as sucrose with gum arabic or tragacanth.

In certain embodiments, the compositions disclosed herein are formulated for rectal administration. In certain embodiments, the compositions disclosed herein are administered as a suppository. In certain embodiments, compositions suitable for rectal administration are prepared by mixing a compound disclosed herein with a suitable non-irritating excipient which is solid at ordinary temperatures but at rectal temperature The lower is a liquid and thus can be dissolved in the rectum to release the drug. In certain embodiments, compositions suitable for rectal administration are prepared by combining the compounds disclosed herein with cocoa butter, glycerinated gelatin, hydrogenated vegetable oil, polyethylene glycol mixtures of different molecular weights or polyethylene glycols. The acid ester is prepared by mixing.

For a method of preparing a variety of pharmaceutical compositions, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Ester, Pa., 18th Ed. (1990).

In certain embodiments, at least 60 percent of the drug is released in 30 minutes using U.S. Pharmacopeia (USP) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as the dissolution medium. In certain embodiments, at least 50-100 percent of the drug is released in 30 minutes using U.S. Pharmacopeia (USP) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as the dissolution medium. In certain embodiments, at least 50-90 percent of the drug is released in 30 minutes using U.S. Pharmacopeia (USP) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as the dissolution medium. In certain embodiments, at least 50-80 percent of the drug is released in 30 minutes using U.S. Pharmacopeia (USP) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as the dissolution medium.

dose

The amount of pharmaceutical composition administered will depend first on the mammal being treated. In the case where the pharmaceutical composition is administered to a human individual, the daily dose is usually determined by the prescribing physician, wherein the dosage generally depends on the age, sex, diet, weight, general health and reaction of the individual, the severity of the individual's symptoms, and the clear adaptation of the treatment. The severity of the condition or condition, the indication for treatment or the severity of the condition, the time of administration, the route of administration, the predisposition of the composition, the rate of excretion, the combination of the drug, and the judgment of the prescribing physician.

In certain embodiments, the dosage is between about 0.001 to about 1000 mg/kg body weight per day. In certain embodiments, the amount of the compound disclosed herein ranges from about 0.5 to about 50 mg/kg/day. In certain embodiments, the amount of the compound disclosed herein is from about 0.001 to about 7 g per day. In certain embodiments, the amount of the compound disclosed herein is from about 0.01 to about 7 g per day. In certain embodiments, the amount of the compound disclosed herein is from about 0.02 to about 5 g per day. In certain embodiments, the amount of the compound disclosed herein is from about 0.05 to about 2.5 g per day. In certain embodiments, the amount of the compound disclosed herein is from about 0.1 to about 1 g per day.

In certain embodiments, the amount of the compound disclosed herein is administered once a day in a single dose. In certain embodiments, the amount of the compound disclosed herein is administered more than once per day in multiple doses. In certain embodiments, the amount of the compound disclosed herein is administered twice daily. In certain embodiments, the amount of the compound disclosed herein is administered three times a day. In certain embodiments, the amount of the compound disclosed herein is administered four times a day. In certain embodiments, the amount of the compound disclosed herein is administered more than four times per day.

Although in other cases larger doses are used without causing any deleterious side effects, in some cases doses below the lower limit of the range mentioned above are more appropriate, by dividing the larger dose into smaller ones. The dose is administered in a single day. The dose varies particular visual IC ₅₀ values of the compounds used. In combination applications where the compound is not monotherapy, a lower amount of the compound can be administered while still having a therapeutic or prophylactic effect.

IX. Combination therapy

In certain embodiments, the compounds disclosed herein are administered in combination with a second therapeutic agent. In certain embodiments, the compounds disclosed herein are administered in combination with surgery and/or radiation therapy.

In certain embodiments, the second therapeutic agent is selected from the group consisting of a cytostatic agent, an anti-angiogenic agent, and an anti-tumor agent. In certain embodiments, the second therapeutic agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; anti-tumor enzymes, topoisomerase inhibitors, procarbazines ), mitoxantrones, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal/anti-hormonal therapeutics, hematopoietic growth factors, aromatase inhibitors, antiestrogens, antiandrogens, cortex Steroids, gonadotropin agonists, microtubule activators, nitrosourea, lipid or protein kinase targets, IMiDs, protein or lipid phosphatase targets, anti-angiogenic agents, Akt inhibitors, IGF-I Inhibitors, FGF3 Regulators, mTOR Inhibitors, Smac mimetics, HDAC Inhibitors, Agents Inducing Cell Differentiation, Bradykinin 1 Receptor Antagonists, Angiotensin II Antagonists, Cyclic Plus Oxygenase inhibitors, heparanase inhibitors, lymphatic mediators, cytokine inhibitors, IKK inhibitors, P38MAPK inhibitors, HSP90 inhibitors, multi-kinase inhibitors, bisphosphonates Rapamycin derivatives, anti-apoptotic pathway inhibitors, apoptosis pathway agonists, PPAR agonists, RAR agonists, Ras isomeric guanidine inhibitors, telomere inhibitors, proteases Inhibitor, metalloproteinase inhibitor, aminopeptidase inhibitor, SHIP activator-AQX-MN100, Humax-CD20 (ofatumumab), CD20 antagonist, IL2-diphtheria toxin fusion or a combination thereof.

In certain embodiments, the second therapeutic agent is selected from the group consisting of ARRY-797, dacarbazine (DTIC), actinomycin C ₂ , C ₃ , D, and F ₁ , cyclophosphamide, melphalan (melphalan), estramustine, maytansinol, rifamycin, streptovaricin, doxorubicin, daunorubicin ), epirubicin, idarubicin, detorubicin, carminomycin, idarubicin, epirubicin, essobi Esorubicin, mitoxantrone, bleomycins A, A ₂ , and B, camptothecin, Irinotecan, Topotecan, 9-amine Glycine base, 10,11-methylenedioxycamptothecin, 9-nitrocamptothecin, bortezomib, temozolomide, TAS103, NPI0052, Combyl Stefandin (combretastatin), Combyl Standin A-2, Combyl Stefandin A-4, calicheamicins, neocarcinostatins, epothilone A, B, C (epothilone s A, B, C) and semisynthetic variants, Herceptin, Rituxan, CD40 antibody, asparaginase, interleukins, interferon, leuprolide ), and pegaspargase, 5-fluorouracil, fluorodeoxyuridine, ptorafur, 5'-deoxy fluorouridine, UFT, MITC, S-1 capecitabine ( Capecitabine), diethylhexyl estradiol, Tamoxifen, toremefine, tolmudex, thymitaq, flutamide, fluoromethylol Anthrone, bicalutamide, finasteride, estradiol, trioxifene, dexamethasone, leuproelinacetate, estramustine (estramustine), droloxifene, medroxyprogesterone, megesterol acetate, aminoglutethimide, testolactone, testosterone , diethylstilbestrol, hydroxyprogesterone, mitomycins A, B and C, methyl Porfiromycin, cisplatin, carboplatin, oxaliplatin, tetraplatin, platinum-DACH, ormaplatin, thalidomide (thalidomide), lenalidomide, CI-973, telomestatin, CHIR258, Rad001, SAHA, Tubacin, 17-AAG, sorafenib, JM -216, podophyllotoxin, epipodophyllotoxin, etoposide, teniposide, Tarceva, Iressa, imatinib (Imatinib), Miltefosine, Perifosine, aminopterin, methotrexate, methopterin, dichloromethanthine (methotrexate), 6-巯嘌呤, thioguanine, zattuoprine, allopurinol, cladribine, fludarabine, pentostatin, 2- Chloromethylamine, deoxycytidine, cytosine arabinoside, cytarabine, azacitidine (azacitidine), 5-azacytosine, gencitabine, azacytosine-arabinoside, vincristine, vinblastine (vinblastine), vinorelbine, leurosine, leurosidine, and vindesine, paclitaxel, taxotere, and docetaxel ( Docetaxel).

In certain embodiments, the second therapeutic agent is selected from the group consisting of a corticosteroid, a non-steroidal anti-inflammatory agent, a muscle relaxant, and combinations thereof with other agents, an anesthetic and combinations thereof with other agents, expectorants, and Combination of other agents, antidepressants, anticonvulsants and combinations thereof; antihypertensives, opioids, topical cannabinoids, capsaicin, betamethasone dipropionate (enhanced and unreinforced) ), betamethasone valerate, clobetasol propionate, prednisone, prednisolone, diflorasone diacetate, c Halobetasol propionate, amcinonide, dexamethasone, dexosimethasone, fluocinolone acetononide, fluocinonide , halocinonide, clocortalone pivalate, dexosimetasone, flurandrenalide, salicylic acid, ibuprofen Ketoprofen, etodolac, diclofenac, meclofenamate sodium, naproxen, piroxicam, celecoxib ( Celecoxib), cyclobenzaprine, baclofen, flubenzate/lidocaine, chloranilide/cyclobenzate, Cyclobenzate/lidocaine/ Ketoprofen, lidocaine, lidocaine/deoxy-D-glucose, prilocaine, EMLA cream (local anesthetic (lidocaine 2.5% and prilocaine 2.5%) Melted mixture), phenfenasin, phenfenazin/ketoprofen/cyclobenzalpine, amitryptiline, doxepin, desipramine , imipramine, amoxapine, clomipramine, nortriptyline, protriptyline, duloxetine, mittazhuo Mirtazepine, nisoxetine, maprotinline, reboxetine, fluoxetine, fluvoxane Mine), carbamazepine, felbamate, lamotrigine, topiramate, tiagabine, oxcarbazepine, carbamezipine ), zonisamide, mexiletine, gabapentin/clonidine, gabapentin/carbamazepine, carbazapine/cyclobenzate, antihypertensives Including clonidine, codeine, loperamide, tramadol, morphine, fentanyl, oxycodone, hydrocodone, levophthine Levophanol, butorphanol, menthol, wintergreen oil, camphor, eucalyptus oil, turpentine; CB1/CB2 ligand, acetaminophen, infliximab, nitric oxide synthesis Enzyme inhibitors, particularly inhibitors that induce nitric oxide synthase, PDE4 inhibitors - similar to the mechanism of Ibudilast (AV-411), CDC-801, JNK inhibitor-CC-401, Combination of TNF/PDE4 inhibitors - CDC-998, IL1 antagonists, such as Anakinra-Kinatt (Anakinra- Kineret), AMG108, target IL-1 (mAb), SHIP activator-AQX-MN100, C5 antagonist, C5a inhibitor, Pexelizumab, pyrimidine synthesis inhibitor, lymphatic mediator, cell Hormone inhibitors, IKK inhibitors, P38MAPK inhibitors, ARRY-797, HSP90 inhibitors, multi-kinase inhibitors, bisphosphonates, PPAR agonists, Cox1 and cox2 inhibitors, anti-CD4 therapy, B-cell inhibitors , COX/LOX dual inhibitors, immunosuppressive agents, iNOS inhibitors, non-steroidal anti-inflammatory drugs (NSAIDs), sPLA2 inhibitors, colchicine, allopurinol, oxypurinol, gold, Ridaura-Auranofin, febuxostat, Puricase, PEG-uricase formulation, Benzbromarone ), long-acting beta-2 agonists (LABAs), salmeterol (Serevent Diskus) and formoterol (Foradil), leukotriene modifiers including montelukast (Singulair) and zafirlukast (Accolate). Inhalation of cromolyn (Intal) or nedocromil (Tilade), theophylline. Short-acting beta-2 agonist, Ipratropium (Atrovent), immunotherapy-(allergy-minmentation injection), anti-IgE monoclonal antibody-Xolair, general DMARD including hydroxychloroquine (Plaquenil), gold compound-Auranofin (Reduca), sulfasalazine (Azulfidine), minocycline (Dynacin, Minocin) and methamine (Rheumatrex) , leflunomide (Arava), azathioprine (Imuran), cyclosporine (Neoral, Sandimmune) and cyclophosphamide (Cytoxan), antibiotics, CD80 antagonists, co-stimulatory factor antagonism Agent, Humax-CD20 (ofatumumab); CD20 antagonist, MEK inhibitor, NFκB inhibitor, anti-B cell antibody, denosumab, receptor activator of specific target nuclear factor kappa B ligand (RANKL) ) mAb. IL17 deactivated antibody, IL-17 receptor antagonist/inhibitor, CTLA inhibitor, CD20 inhibitor, soluble VEGFR-1 receptor, anti-VEGFR-1 receptor antibody, anti-VEGF antibody, integrin receptor antagonist, Selectin inhibitors, P-selectin and E-selectin inhibitors, phospholipase A2 inhibitors, lipoxygenation inhibitors, RANKL and RANK antagonists/antibodies, osteoproteger antagonists, lymphotoxin inhibitors, B - lymphocyte stimulator, MCP-1 inhibitor, MIF inhibitor, CD2, CD3, CD4, CD25, CD40 and CD40 ligand CD152 (CTLA4) inhibitor, macrolide (Macrolide) immunosuppressant, nucleus Selective inhibitors of nucleotide metabolism, chemotactic inhibitors, CXC receptors and CXC ligand inhibitors, Chemokine antagonists, leukocyte chemotactic inhibitors, adhesion molecule blockers, selectins Lymphocyte functional antigen-1 (LFA-1, CD11a) antagonist, very late antigen-4 (VLA-4) antagonist, matrix metalloproteinase inhibitor, elastase inhibitor, cathepsin inhibitor.

In certain embodiments, the second therapeutic agent is selected from the group consisting of a beta-blocker, a carbonic anhydrase inhibitor, an alpha- and beta-adrenergic antagonist, including an alpha 1-adrenergic antagonist, an alpha 2 agonist, a miotic drug , prostate analogs, corticosteroids and immunosuppressive agents. In certain embodiments, the second therapeutic agent is selected from the group consisting of timolol, betaxolol, levobetaxolol, carteolol, levobunolol (levobunolol), propranolol, brinzolamide, dorzolamide, nipradil, iopidine, brimonidine ), pilocarpine, adrenaline, latanoprost, travoprost, bimatoprost, unoprostone, dexamethasone, prednisone, Methylprednisolone, azathioprine, cyclosporine and immunoglobulin.

In certain embodiments, the second therapeutic agent is selected from the group consisting of a corticosteroid, an immunosuppressive agent, a prostate analog, and an antimetabolite. In certain embodiments, the second therapeutic agent is selected from the group consisting of dexamethasone, prednisone, methylprednisolone, azathioprine, cyclosporine, immunoglobulin, latanoprost, trovoprost Travoprost, bimatoprost, unoprostone, infliximab, rituximab, methotrexate, non-steroidal anti-inflammatory agents, muscle Relaxant and its combination with other agents, anesthetics and combinations thereof with other agents, combination of expectorants and other agents, antidepressants, anticonvulsants and combinations thereof; antihypertensive drugs, opioids, Topical cannabinoids, and other agents, such as capsaicin, betamethasone dipropionate (enhanced and unreinforced), betamethasone valerate, clobetasol propionate, prednisone, methylprednisolone, diacetic acid Diflurazon, halobutazone propionate, ansinide, dexamethasone, dexemethone, acetone fluocinolone, fluocinolone, flufensulfonate, coca Dintermate, fluorohydrogen, salicylic acid, isoprofen, ketoprofen, etodo , diclofenac, meloprofen, naproxen, piroxicam, celecoxib, Cyclobenzine, bucklefin, Cyclobenzapine/lidocaine, bucklefin/cyclobenzate, ring Benzapride/lidocaine/ketoprofen, lidocaine, lidocaine/deoxy-D-glucose, prilocaine, EMLA cream (local anesthetic (lidocaine 2.5% and prilocaine) (prilocaine) 2.5%) of the eutectic mixture), penfenazin, piperphenazine/ketoprofen/cyclobenzalazine, amitriptyline, doxepin, desipramine, imipramine , amoxapine, clomipramine, nortriptyline, protriptyline, duloxetine, mitazodine, nisoloxine, maprotiline, reboxetine, fluoxetine, flu Samin, carbazapine, felamine, lamotrigine, topiramate, tiagabine, oxcarbazepine, carbamazepine, zonisamide, mexiletine, gabapentin/ clonidine, gabapentin/carbazide Carbazide/cyclobenzate, antihypertensives including clonidine, codeine, loperamide, tramadol, morphine, fentanyl, oxycodone, hydrocodone, levonorol, buto Morino, mint Brain, wintergreen oil, camphor, eucalyptus oil, turpentine; CB1/CB2 ligand, acetaminophen, infliximab, nitric oxide synthase inhibitor, especially inhibitors that induce nitric oxide synthase; And other agents, such as capsaicin. PDE4 inhibitor - similar to the mechanism of Ibudilast (AV-411), CDC-801, JNK inhibitor-CC-401, combined TNF/PDE4 inhibitor-CDC-998, IL1 antagonist such as Anna Anakinra-Kineret, AMG108, target IL-1 (mAb), SHIP activator-AQX-MN100, C5 antagonist, C5a inhibitor, Pexelizumab, pyrimidine synthesis inhibition Agents, lymphatic mediators, cytokine inhibitors, IKK inhibitors, P38MAPK inhibitors, ARRY-797, HSP90 inhibitors, multi-kinase inhibitors, bisphosphonates, PPAR agonists, Cox1 and cox2 inhibitors, antibiotics CD4 therapy, B-cell inhibitor, COX/LOX dual inhibitor, immunosuppressive agent, iNOS inhibitor, NSAID, sPLA2 inhibitor, colchicine, allopurinol, oxypurinol, gold, rudol La-Ginofin, febuxostat, Puricase, PE G-uricase, benzolamone, long-acting beta-2 agonist (LABAs), Serevent Diskus and Foradil, leukotriene modifiers include Singulair and Accort. Inhalation of Intal or Tilade, theophylline. Short-acting β-2 agonist, Atrovent, Immunotherapy-(Allergy-Desensitization Injection), Anti-IgE monoclonal antibody-Cymol, General DMARD including hydroxychloroquine (Plaquenil), gold compound - auranofin (Reduca), sulfasalazine (Azulfidine), minocycline (Dynacin, Minocin) and methamine (Rheumatrex), levamite (Arava), azathioprine (Imuran) , cyclosporine (Neoral, Sandimmune) and cyclophosphamide (Cytoxan), antibiotics, CD80 antagonists, costimulatory factor antagonists, Humax-CD20 (ofatumumab); CD20 antagonists, MEK inhibitors, NFκB inhibitors, Anti-B cell antibody, mononorm, mAb of specific receptor nuclear factor kappa B ligand receptor activator (RANKL), IL17 deactivated antibody, IL-17 receptor antagonist/inhibitor, CTLA inhibitor, CD20 inhibitor, soluble VEGFR-1 receptor, anti-VEGFR-1 receptor antibody, anti-VEGF antibody, integrin receptor antagonist, selectin inhibitor, P-selectin and E-selectin inhibitor, phospholipase A2 Inhibitors, fat oxime inhibitors, RANKL and RANK antagonists/antibodies, osteoproteger antagonists, lymphotoxin inhibitors, B-drug Bar cell stimulator, MCP-1 inhibitor, MIF inhibitor, CD2, CD3, CD4, CD25, CD40 and CD40 ligand CD152 (CTLA4) inhibitor, macrolide (Macrolide) immunosuppressant, nucleoside Selective inhibitors of acid metabolism, chemotactic inhibitors, CXC receptors and CXC ligand inhibitors, chemokine antagonists, leukocyte chemotactic inhibitors, adhesion molecule blockers, selectin lymphocyte functional antigens -1 (LFA-1, CD11a) antagonist, very late antigen-4 (VLA-4) antagonist, matrix metalloproteinase inhibitor, elastase inhibitor, cathepsin inhibitor.

In certain embodiments, the second therapeutic agent is selected from the group consisting of insulin, insulin derivatives and peptidomimetics, insulin secretagogues, insulin sensitizers, biguanides, alpha-glucosidase inhibitors, insulinotropic sulfonyl ureas Somatic ligand, protein tyrosine phosphatase-1B (PTP-1B) inhibitor, GSK3 (glycosidase kinase-3) inhibitor, GLP-1 (glycoglycin peptide-1), GLP- 1 analog, DPPIV (dipeptidyl peptidase IV) inhibitor, RXR ligand sodium-dependent glucose co-transporter inhibitor, glycophosphorylase A inhibitor, AGE fragment, PPAR regulator, LXR and FXR Regulator, non-glitazone type PPARS agonist, selective glucocorticoid antagonist, metformin, Glipizide, glyburide, Mare pancreas (Amaryl), meglitinides, 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 of the compound l9 of the patent application WO03/043985 -[5-methyl-2-(4-trifluoromethyl-phenyl)-oxazole-4-ylmethoxy]-benzenesulfonyl}2,3-dihydro-1H-indole-2 - Carboxylic acid and GI-262570.

Set

The compounds, compositions, and methods described herein provide a kit for treating a condition, such as the conditions described herein. Such kits comprise a compound, a plurality of compounds or compositions described herein in a container, and optionally include instructions for using the kit in accordance with various methods and means described herein. The kits may also contain information such as scientific literature references, package insert materials, clinical trial results, and/or an overview of such information and analogs thereof that indicate or establish the activity and/or advantages of the composition, And/or describe other information available to the administration, administration, side effects, drug interactions, or health care providers. This information is based on the results of various studies, such as studies using in vivo models of experimental animals and studies based on human clinical trials. The kits described herein are provided, marketed, and/or extended to health care providers, including physicians, nurses, pharmacists, formulary officials, and the like. Also, in some embodiments, the kit can be sold directly to the consumer.

Instance General Illustrative Procedure for the Synthesis of Sulfonamide

Procedure A: Anhydrous triethylamine (5 eq) was added to a solution of EtOAc (1 EtOAc). To the solution was added sulfonium chloride (1 eq) and the solution was stirred at room temperature for 16 hours. The solvent was evaporated and the residue was purified on EtOAc EtOAc.

Procedure B: To a stirred solution of the amine (1 eq) in dry pyridine (5 ml / mmol) was added sulphonium chloride (1-5 eq). The reaction mixture was stirred at 40 ° C for 48 hours. The reaction mixture was partitioned between water and EtOAc. The organic layer was washed with brine, dried (MgSO ₄ The residue was purified on cerium oxide by flash column chromatography.

Procedure C (substitution of iodine atom): heating at 115 ° C in a microwave reactor containing 1 eq. aryl iodide, 1.5 equivalents Acid or The acid ester, 0.25 eq. PdCl ₂ (dppf) ‧ DCM and 10 eq. of a suspension of anhydrous K ₂ CO ₃ powder in a deoxygenated mixture of dioxane and water (3 1) for 60 minutes. It was extracted with aqueous NH ₄ Cl/THF, and the organic portion was dried using Na ₂ SO ₄ . Purification by flash column chromatography (Si, EtOAc / hexane or CHCl ₃ / MeOH) the crude reaction product. The yield is 20-40%.

Procedure D (Synthesis of N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-2-(alkylamino)ethanesulfonamide): 2-Chloro-ethanesulfonium chloride (0.1 ml, 1 mmol) was added to 5,6-difluoro- N ¹ -(2-fluoro-4-iodophenyl)benzene-1,2-diamine (0.364 g, 1 mmol of) and triethylamine (0.28ml, 2mmol) in CH ₂ Cl ₂ (5ml) in the solution and the reaction mixture was stirred for 16 hours at room temperature. This was then treated with excess amine (10 eq) as a solution or neat liquid. The reaction mixture was stirred for a further 6 hours at room temperature. The reaction mixture was diluted with CH ₂ Cl ₂ (10 mL) and water (10 mL). The organic layer was washed successively with dilute HCl (2×20 mL, 2N) and sat. NaHCO ₃ (2×10 mL). Next, this CH ₂ Cl ₂ layer was dried (MgSO ₄ ) and evaporated to give a crude material. This impure product was purified under preparative HPLC conditions to obtain the pure product (yield 50-60%).

Example 1: N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonate Guanamine

Step A: butyl cyclopropane sulfonate

The cyclopropane sulfonium chloride (5 g, 35 mmol, 1 eq) was dissolved in EtOAc (20 mL). EtOAc (EtOAc) The mixture was slowly warmed at room temperature and stirred overnight. The solvent was removed under reduced pressure and the resulting white solid was dissolved in CHCl _3. In water, the organic phase was washed with brine and dried (MgSO ₄₎ and concentrated to give an oil (4.8g, 24.9mmol, 71%) . _{^{1 H NMR (300MHz, CDCl 3}} ) δ4.25 (t, 2H), 2.46 (m, 1H), 1.74 (m, 2H), 1.45 (m, 2H), 1.25 (dd, 2H), 1.09 (dd, 2H), 0.93 (t, 3H).

Step B: 1-Allylcyclopropane-1-sulfonic acid butyl ester

To a solution of 1-butyl cyclopropanesulfonate (4.8 g, 24.9 mmol) in THF at -78 ° C under nitrogen atmosphere, butyl lithium solution (15.6 m, 24.9 mmol, 1.6 M, THF) and Allyl iodide (24.9 mmol). The reaction mixture was stirred at -78 °C for 2 hours and at room temperature for 3 hours. The volatiles were evaporated off under reduced pressure and with CH ₂ Cl ₂ (100ml) and extracted residue. The extract was washed with water, dried (MgSO ₄₎ and evaporated. Via a silicon dioxide gel chromatography technique (eluent: hexane / CH ₂ Cl ₂₎ was obtained residue was purified title product as a colorless oil of (3.75g, 69.0%). ¹ H NMR (300MHz, CDCl ₃ ) δ 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), 0.93 (m, 5H).

Step C: 1-Allylcyclopropane-1-sulfonic acid potassium

A mixture of 1-butyl-cyclopropanesulfonic acid 1-butyl ester (3.75 g, 17.2 mmol) and potassium thiocyanate (1.7 g, 17.2 mmol) in DME (20 mL) The volatiles were evaporated to give a crude sulfonic acid salt (3.44 g, quantitative) which was dried under vacuum at 50 ° C for 16 hours. This crude product was used in the next reaction without further purification. ¹ H NMR (CDCl ₃ ) δ 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: 1-allyl cyclopropane-1-sulfonyl chloride

A solution of potassium 1-allyl cyclopropane-1-sulfonate (3.44 g, 172 mmol), sulfinium chloride (10 ml) and DMF (5 drops) was refluxed at 60 ° C for 16 hours. Under reduced pressure and the volatiles were evaporated in CH ₂ Cl ₂ (50ml) and extracted residue. The extract was washed with water, dried and used without further purification (MgSO ₄₎ and evaporated to give a yellow gummy oil of the crude product was washed with hexane crude product was used in the next reaction (2.7g, 15mmol, 87% ). ¹ H NMR (300 MHz, CDCl ₃ ) δ 5.728 (m, 1H), 5.91 (t, 2H), 2.9 (d, 2H), 0.756 (m, 2H), 0.322 (m, 2H).

Step E: 1-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)benzene-1,2-diamine is reacted with 1-allyl cyclopropane-1-sulfonyl chloride Obtain the desired product. m / z =507[M-1] ^- .

Step F: N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonate Guanamine

1-Allyl-N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)cyclopropane-1-sulfonamide (0.77 g, 1.52 mmol) And 4-methylmorpholine N-oxide (0.18 g, 1.52 mmol) was dissolved in THF (50 mL). Osmium tetroxide was added at room temperature (0.152mmol, 0.965mL, 4% in H ₂ O) is added and the mixture stirred for 16 hours at room temperature the reaction. Was added with EtOAc, the organic phase was washed with water, dried (MgSO ₄₎ and concentrated under reduced pressure. The residue was purified with EtOAc EtOAc (EtOAc) ¹ H NMR (300 MHz, CDCl ₃ + D ₂ O) δ 7.38 (dd, J = 1.8 & 10.5 Hz, 1H), 7.36 (ddd, J = 2.4, 5.1 & 9.3 Hz, 1H), 7.25 (d, J) = 8.7 Hz, 1H), 7.02 (dd, J = 9.0 & 17.7 Hz, 1H), 6.27 (dt, J = 3.0, 8.7 & 17.4 Hz, 1H), 3.92 (m, 1H), 3.54 (dd, J = 3.9 & 11.1 Hz, 1H), 3.39 (dd, J = 6.6 & 11.1 Hz, 1H), 2.16 (dd, J = 9.6 & 15.9 Hz, 1H), 1.59 (d, J = 14.1 Hz, 1H), 1.41 ( m, 1H), 1.26 (m, 1H), 0.83 (m, 2H); m / z = 542 [M-1] ^- .

Example 2: (S)-N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane -1-sulfonamide

The pure S isomer was obtained by fractional HPLC separation of the racemic mixture (Example 13). ¹ H NMR (300 MHz, CDCl ₃ + D ₂ O) δ 7.38 (dd, J = 1.8 & 10.5 Hz, 1H), 7.36 (ddd, J = 2.4, 5.1 & 9.3 Hz, 1H), 7.25 (d, J) = 8.7 Hz, 1H), 7.02 (dd, J = 9.0 & 17.7 Hz, 1H), 6.27 (dt, J = 3.0, 8.7 & 17.4 Hz, 1H), 3.92 (m, 1H), 3.54 (dd, J = 3.9 & 11.1 Hz, 1H), 3.39 (dd, J = 6.6 & 11.1 Hz, 1H), 2.16 (dd, J = 9.6 & 15.9 Hz, 1H), 1.59 (d, J = 14.1 Hz, 1H), 1.41 ( m, 1H), 1.26 (m, 1H), 0.83 (m, 2H); m / z = 542 [M-1] ^- .

Example 3: (R)-N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane -1-sulfonamide

The pure R isomer was obtained by fractional HPLC separation of the racemic mixture (Example 13). ¹ H NMR (300 MHz, CDCl ₃ + D ₂ O): δ 7.38 (dd, J = 1.8 & 10.5 Hz, 1H), 7.36 (ddd, J = 2.4, 5.1 & 9.3 Hz, 1H), 7.25 (d, J=8.7 Hz, 1H), 7.02 (dd, J=9.0 & 17.7 Hz, 1H), 6.27 (dt, J=3.0, 8.7 & 17.4 Hz, 1H), 3.92 (m, 1H), 3.54 (dd, J =3.9 & 11.1 Hz, 1H), 3.39 (dd, J = 6.6 & 11.1 Hz, 1H), 2.16 (dd, J = 9.6 & 15.9 Hz, 1H), 1.59 (d, J = 14.1 Hz, 1H), 1.41 (m, 1H), 1.26 (m, 1H), 0.83 (m, 2H); m/z = 542 [M-1] ^- .

Example 4: 1-(2,3-Dihydroxy-propyl)-cyclopropanesulfonic acid [3,4,6-trifluoro-2-(4-fluoro-2iodo-phenylamino)-phenyl] - guanamine

Step A 1-Allyl-cyclopropanesulfonic acid [3,4,6-trifluoro-2-(2-fluoro-4-iodo-phenylamino)phenyl]-decylamine

Reaction of 1-allyl-cyclopropanesulfonium chloride with 3,5,6-trifluoro-N ¹ -(2-fluoro-4-iodophenyl)benzene-1,2-diamine according to the general procedure B Obtain the title product. _{^{1 H NMR (CDCl 3, 300MHz}} ) δ7.41 (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).

Step B 1-(2,3-Dihydroxypropyl)-N-(3,4,6-trifluoro-2-(2-fluoro-4-iodophenylamino)phenyl)cyclopropane-1- Sulfonamide

1-Allyl-cyclopropanesulfonic acid [3,4,6-trifluoro-2-(2-fluoro-4-iodo-phenylamino)-phenyl]-decylamine (110 mg, 0.21 mmol) And 4-methylmorpholine N-oxide (24.6 mg, 0.21 mmol) was dissolved in THF (8 mL). Osmium tetroxide was added at room temperature (0.021mmol, 0.153mL, 4% in H ₂ O) is added and the mixture stirred for 16 hours at room temperature the reaction. Was added with EtOAc, the organic phase was washed with water, dried (MgSO ₄₎ and concentrated under reduced pressure. The residue was purified with EtOAc EtOAc (EtOAc) ¹ H NMR (CDCl ₃ , 300 MHz) δ 7.39 (dd, J = 1.5 & 10.6 Hz, 1H), 7.29 (d, J = 8.8 Hz, 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 & 11.4 Hz, 1H), 3.53 (dd. J = 6.7 & 11.2 Hz, 1H), 2.50 (dd, J = 10.0 & 16.1 Hz, 1H), 1.6 (m, 1H), 1.46 (m, 1H), 1.28 (m, 1H), 1.20 (m, 2H), 0.92 (m, 2H); m /z =559[M-1] ^- .

Example 5: (S)-1-(2,3-dihydroxypropyl)-N-(3,4,6-trifluoro-2-(2-fluoro-4-iodophenylamino)phenyl) Cyclopropane-1-sulfonamide

The pure S isomer was obtained by fractional HPLC separation of the racemic mixture (Example 52). ¹ H NMR (CDCl ₃ , 300 MHz) δ 7.39 (dd, J = 1.5 & 10.6 Hz ₅ 1H), 7.29 (d, J = 8.8 Hz, 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 & 11.4 Hz ₅ 1H), 3.53 (dd, J = 6.7 & 11.2 Hz, 1H), 2.50 (dd, J = 10.0 & 16.1 Hz, 1H), 1.6 (m, 1H), 1.46 (m, 1H), 1.28 (m, 1H), 1.20 (m, 2H), 0.92 (m, 2H); m / z =559[M-1] ^- .

Example 6: (R)-1-(2,3-dihydroxypropyl)-N-(3,4,6-trifluoro-2-(2-fluoro-4-iodophenylamino)phenyl) Cyclopropane-1-sulfonamide

The pure R isomer was obtained by fractional HPLC separation of the racemic mixture (Example 52). ¹ H NMR (CDCl ₃ , 300 MHz) δ 7.39 (dd, J = 1.5 & 10.6 Hz, 1H), 7.29 (d, J = 8.8 Hz, 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 & 11.4 Hz, 1H), 3.53 (dd, J = 6.7 & 11.2 Hz, 1H), 2.50 (dd, J = 10.0 & 16.1 Hz, 1H), 1.6 (m, 1H), 1.46 (m, 1H), 1.28 (m, 1H), 1.20 (m, 2H), 0.92 (m, 2H); m / z =559[M-1] ^- .

Example 7: N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonate Guanamine

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

1-Allyl-cyclopropanesulfonium chloride and 5,6-difluoro-N1-(2-fluoro-4-iodophenyl)-3-methoxybenzene-1,2- according to the general procedure B The diamine reaction is carried out to obtain the title product. ¹ H NMR (CDCl ₃ , 300 MHz) δ 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, 1H), 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 BN-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide

1-Allyl-N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)cyclopropane-1-sulfonamide ( 97 mg, 0.18 mmol) and 4-methylmorpholine N-oxide (21 mg, 0.18 mmol) were dissolved in THF (8 mL). Osmium tetroxide was added at room temperature (0.018mmol, 0.13mL, 4% in H ₂ O) is added and the mixture stirred for 16 hours at room temperature the reaction. Was added with EtOAc, the organic phase was washed with water, dried (MgSO ₄₎ and concentrated under reduced pressure. The residue was purified with EtOAc EtOAc (EtOAc) _{^{1 H NMR (CDCl 3, 300MHz}} ) δ7.38 (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 & 11.4 Hz, 1H), 6.43 (m, 1H), 4.06 (m, 1H), 3.89 (s, 3H), 3.63 (dd, J = 3.7 & 11.1 Hz, 1H), 3.49 (dd, J = 6.4 & 11.1 Hz, 1H), 2.3 (dd, J = 9.7 & 16.1 Hz, 1H), 1.77 (dd, J = 1.9 & 16.0 Hz, 1H), 1.37 (m, 1H), 1.25 (m, 1H) , 1.21 (m, 2H), 0.86 (m, 2H); m/z = 571 [M-1] ^- .

Example 8: (S)-N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Hydroxypropyl)cyclopropane-1-sulfonamide

The pure S isomer was obtained by a palmitic HPLC separation of the racemic mixture (Example 55). _{^{1 H NMR (CDCl 3, 300MHz}} ) δ7.38 (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 & 11.4 Hz, 1H), 6.43 (m, 1H), 4.06 (m, 1H), 3.89 (s, 3H), 3.63 (dd, J = 3.7 & 11.1 Hz, 1H), 3.49 (dd, J = 6.4 & 11.1 Hz, 1H), 2.3 (dd, J = 9.7 & 16.1 Hz, 1H), 1.77 (dd, J = 1.9 & 16.0 Hz, 1H), 1.37 (m, 1H), 1.25 (m, 1H) , 1.21 (m, 2H), 0.86 (m, 2H); m/z = 571 [M-1] ^- .

Example 9: (R)-N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Hydroxypropyl)cyclopropane-1-sulfonamide

The pure R isomer was obtained by a palmitic HPLC separation of the racemic mixture (Example 55). _{^{1 H NMR (CDCl 3, 300MHz}} ) δ7.38 (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 & 11.4 Hz, 1H), 6.43 (m, 1H), 4.06 (m, 1H), 3.89 (s, 3H), 3.63 (dd, J = 3.7 & 11.1 Hz, 1H), 3.49 (dd, J = 6.4 & 11.1 Hz, 1H), 2.3 (dd, J = 9.7 & 16.1 Hz, 1H), 1.77 (dd, J = 1.9 & 16.0 Hz, 1H), 1.37 (m, 1H), 1.25 (m, 1H) , 1.21 (m, 2H), 0.86 (m, 2H); m/z = 571 [M-1].

Example 10: N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methylphenyl)-1-(2,3-dihydroxypropyl) ring Propane-1-sulfonamide :

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

Slowly batchwise to a cooled (-5 ° C) solution of 3,4,5-trifluorobenzaldehyde (7.0 g, 43.75 mmol) in a mixture of THF and water (50 mL, 9:1) over 30 min. NaBH ₄ (1.662 g, 43.75 mmol) was added. The reaction mixture was allowed to reach room temperature over 2 h and was poured EtOAc EtOAc EtOAc (EtOAc) The oily layer was extracted into CH ₂ Cl ₂ (250ml) and the organic layer was with water (200ml), dried (MgSO ₄₎ and evaporated. The crude product obtained (7.08 g, quantitative) was taken to the next step without further purification.

step Step B: 5-(Bromomethyl)-1,2,3-trifluorobenzene:

In the ₂ Cl ₂ (150ml) was slowly added thionyl acyl bromine (6.16ml, 80mmol) in CH ₂ Cl ₂ (50ml) solution of (3,4,5-trifluorophenyl) methanol (40mmol) in CH of Solution. The reaction mixture was stirred at room temperature for 16 hours and poured into ice water (200 ml). The organic layer was separated and washed with saturated NaHCO ₃ (2 × 200ml), water (200ml), dried (MgSO ₄₎ and evaporated to give the corresponding bromo compound as a pale yellow oil of (quantitative yield). This crude product was taken to the next reaction without further purification.

Step C: 1,2,3-Trifluoro-5-methylbenzene:

The bromo compound (40 mmol) was combined with triethyl decane (48 mmol) and the reaction mixture was worked up in small portions of solid PdCl ₂ (4 mmol). After a few minutes, a violent exothermic reaction occurred, carefully placing the contents of the flask back by placing a reflux condenser. The reaction mixture was further stirred at room temperature for 6 hours and the contents were allowed to stand for 16 hours. The crude liquid product was then carefully decanted and passed directly to the next reaction without further purification. It is assumed that the reaction is carried out in quantitative yield.

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

At 0-5 deg.] C 1,2,3-trifluoro-5-methyl-benzene (40mmol) was added to concentrated H ₂ SO ₄ (50ml). Subsequently, the reaction mixture was slowly concentrated _{HNO 3 (3.39ml, 48.44mmol, 90} %) process, while maintaining the internal temperature below 20 ℃. For 16 h and poured onto ice (300g), in _{CH 2 Cl 2 (2 × 125ml} ) and extracted oily layer was stirred at room temperature. With water (2 × 200ml), the organic layer was washed with brine (200ml) and dried (MgSO ₄₎ and evaporated to give the crude product was purified by flash surgery through the silicon dioxide gel chromatography to obtain a crude product of the title product (6.5 g of, 85%). _{1H-NMR (300MHz, CDCl 3} ): 56.96 ( septet, 1H), 2.39 (s, 3H). ¹⁹ F NMR (CDCl ₃ ): δ-128.18, -141.50, -159.05.

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

2-Fluoro-4-iodoaniline and 1,2,3-trifluoro-5-methyl-4-nitrobenzene were reacted using the conditions described in Example 1 (Step A) to afford the title compound. MH ⁺ : 407.9.

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

2,3-Difluoro-N-(2-fluoro-4-iodophenyl)-5-methyl-6-nitroaniline was reacted using the conditions described in Example 1 (Step B) to give the title compound. MH ⁺ : 377.4.

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

1-Allyl-cyclopropanesulfonium chloride (142 mg, 142 mg) and 5,6-difluoro-N1-(2-fluoro-4-iodophenyl)-3-methylbenzene were used according to the general procedure B. The title product (100 mg, 47%); m/z = 521 [M-1].

Step H: N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methylphenyl)-1-(2,3-dihydroxypropyl) ring Propane-1-sulfonamide:

1-Allyl-N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methylphenyl)cyclopropane-1-sulfonamide (150 mg , 0.29 mmol) and 4-methylmorpholine N-oxide (33 mg, 0.29 mmol) were dissolved in THF (50 mL). Osmium tetroxide was added at room temperature (0.029mmol, 0.18mL, 4% in H ₂ O) is added and the mixture stirred for 16 hours at room temperature the reaction. Add EtOAc, the organic phase was washed with water, dried (MgSO ₄₎ and concentrated under reduced pressure. The residue was purified with EtOAc EtOAc (EtOAc) ¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.07 (m, 1H), 6.97 (brm, 2H), 6.84 (m, 2H), 6.60 (brm, 2H), 3.98 (brm, 1H) , 3.58 (m, 1H), 3.43 (m, 1H), 3.20 (d, J = 3.9 Hz, 1H), 2.42 (s, 3H), 2.31 (dd, J = 9.9 & 15.6 Hz, 1H), 2.01 ( Br t,1H), 2.31 (dd, J=9.9 & 15.6 Hz, 1H), 1.66 (dd, J=2.1 & 15.9 Hz, 1H), 1.52 (m, 1H), 1.40 (m, 1H), 0.91 ( m, 2H).

In vitro biological activity

Example 11: IC ₅₀ Data generation

Preparation of materials and reagents: Human GST-MEK1 and the constitutively active dual gene GST-MEK1 ^CA (with mutations Ser218Asp and Ser222Asp) were sub-selected from the wild-type human MEK1 cDNA in the yeast expression vector pGEM4Z (Promega, Madison, WI). . GST-MEK1 ^CA was visualized in Escherichia coli and partially purified using glutathione Sepharose 4B affinity resin (Amersham Pharmacia Biotech, Piscataway, NJ). The ERK2 dual gene was subcloned from the MAPK2/Erk2 cDNA (wild type) (Upstate Biotechnology, Inc., Waltham, MA) in pUSEamp in the vector pET21a (Novagen, Madison, WI) to generate an N-terminal histidine label. Mouse ERK2 dual gene. ERK2 was expressed and purified to homogeneity [Zhang, 1993 #33]. Myelin basic protein (MBP) was purchased from Gibco BRL (Rockville, MD). EasyTides adenosine 5'-triphosphate (ATP) ([γ- ³³ P]) (NEN Perkin Elmer, Wellesley, MA) is the radiolabeling source for all kinase reactions. Activated Raf-1 (truncated) and activated MAP kinase 2/ERK2 were purchased from Upstate (Lake Placid, NY). 4-20% Criterion Precast gel was purchased from Bio-Rad (Hercules, CA).

Determination of activity: The compound was diluted from a stock solution of dimethyl sulfoxide (DMSO) to 1 × HMNDE (20mM HEPES pH 7.2,1mM MgCl 2, 100mM NaCl, 1.25mM DTT, 0.2mM EDTA) in. A typical 25 ml assay contains 0.002 Nemo MEK1 ^CA , 0.02 Nemo ERK2, 0.25 Nemo MBP, 0.25 Nemo unlabeled ATP, and 0.1 μCi [γ ³³ P]ATP. The screening assay consists essentially of four additions. 5 μl of the diluted compound was dispensed into a 96-well assay plate. Then 10 μl of a 2.5× enzyme mixture (MEK1 ^CA and ERK2 only) was added to each well, followed by pre-incubation for 30 minutes at ambient temperature. Then 10 μl of a 2.5× substrate mixture (labeled and unlabeled ATP plus MBP) was added, followed by incubation at ambient temperature for 60 minutes. Finally, 100 μl of 10% trichloroacetic acid (TCA) was added and incubated at room temperature for 30 minutes to stop the reaction and precipitate the radiolabeled protein product. The reaction product was collected on 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 for 30 minutes at room temperature. A back seal was manually applied and 40 μl of scintillation cocktail was dispensed into each well. A top seal was applied and the plates were counted in a TopCount (2 sec/well).

For some experiments, a truncated version of MEK that requires activation by Raf kinase was used.

real Example 12: Generation of EC50 data

The effect of the compound in the cells is determined by Western blotting of the phosphorylated ERK. MDA-MB-231 breast cancer cells were plated in 48-well plates at 20,000 cells per well and grown in a 37° humidified CO ₂ incubator. The next day, growth medium (DMEM + 10% fetal bovine serum) was removed and replaced with starvation medium (DMEM + 0.1% fetal bovine serum). The cells were cultured for 16 hours in starvation medium and then treated with a series of concentrations of compound for 30 minutes. After incubation with the compound, the cells were stimulated with 100 ng/ml EGF for 5 minutes. The cells were then lysed and analyzed by Western blotting using monoclonal antibodies raised against phosphorylated ERK. The secondary antibody was combined with a near-infrared dye to amplify the signal and detect it on a Licor Odyssey scanner. The signal intensity was quantified and this data was used to generate a dose response curve and perform an EC50 calculation.

Living m biological activity

Example 13

The compounds and compositions described herein are useful for treating or preventing one or more conditions including, but not limited to, cancer, inflammatory bowel disease (IBD), psoriasis, and rheumatoid arthritis (RA). The compounds and compositions described herein can also be used to treat or prevent one or more conditions including, but not limited to, cancer, IBD, psoriasis, and RA, once or twice daily.

A live d test of a compound of the following structure (Compound A, prepared as described herein) is illustrated in this example:

Human tumors were implanted into nu/nu mice. Once the tumor size was about 100 mm ³ , Compound A was administered orally for 14 days. Tumor growth inhibition rate (TGI) was determined after 14 days of treatment, expressed as the rate of decrease in tumor size of the treatment group relative to the vehicle control group. The time to end (TTE) was calculated based on the time at which the tumor reached the specified endpoint volume or the last day of the study, whichever occurs first. Treatment outcomes were determined as a percentage of tumor growth delay (% TGD), which was defined as the percentage increase in mean TTE of treated mice compared to vehicle treated control mice. The regression response of the animals was also monitored. The pERK content in tumors and brain was determined by Western blotting and correlated with the plasma content of Compound A for pharmacodynamic/pharmacokinetic studies. Several tumor models were evaluated at different doses and dose schedules. Statistically significant % TGD was shown at 25 or 50 mg/kg once daily (QD) in A375 melanoma tumors, Colo205 colon cancer tumors, and A431 epidermoid tumors. For oral administration at 25 mg/kg QD in these tumor models as well as in HT29 colon cancer tumors, a statistically significant TGI was observed. The effects of different dosing regimens were evaluated in A375 allografts. Although 100 mg/kg of Compound A administered orally once every two days showed a statistically significant % TGD (91%), it was not as effective as QD treatment of 25 mg/kg (143% TGD) or 50 mg/kg (233% TGD). As measured by %TGI, twice daily (BID) administration is also more effective than QD administration. Administration of 12.5 mg/kg BID yielded 79.5% TGI compared to 51.7% of Compound A's 25 mg/kg QD. Administration of 25 mg/kg BID produced 110.1% TGI compared to 69.9% of 50 mg/kg QD. A pharmacokinetic/pharmacokinetic study in Colo205 xenografts showed inhibition of pERK formation in tumors, while minimal inhibition was observed in the brain, indicating potent antitumor activity with limited CNS permeability.

Compound A is a potent MEK1/2 inhibitor that inhibits tumor cell growth in vitro and in vivo . The BRAF status in fixation-dependent growth but not fixation-dependent growth or in allogeneic transplantation determines sensitivity to growth inhibition by the compound. Maintaining sufficient MEK inhibition during the dosing interval appears to be more important than the peak content because of the greater efficacy under more frequent dosing. Compound A has a favorable pharmacokinetic profile in humans and the estimated therapeutic dose in humans based on xenograft results is 20-40 mg/day.

Example 13A: Inhibition of cancer cell growth (GI ₅₀ )

Fixation-dependent growth inhibition was measured using CellTiterGlo reagent after treatment of cells grown in 384-well plates with Compound A for 48 hours. Fixation independent growth assay MTS (methane sulfonate) reagent was used after 7 days of treatment on cells grown in 0.15% agarose or grown on unbound plates (A431). The growth inhibition value (GI ₅₀ ) is shown in the table below.

Example 13B: Anti-tumor allograft activity

Female nu/nu mice were transplanted with A375 melanoma, Colo205 colon tumor, A431 epidermoid tumor or HT-29 colon tumor cells, and the cells were grown to 100-200 mm ³ . Compound A or vehicle (25 mg/kg, 50 mg/kg or 100 mg/kg) was administered orally once a day for 14 days. The mean tumor volume of the vehicle and treatment groups is plotted and shown in Figure 1.

Example 13C: Tumor growth inhibition rate (TGI) 25 mg/kg QD

The tumor growth inhibition rate of the group treated with 25 mg/kg of Compound A was calculated for the designated tumor xenograft. The tumor growth inhibition rate was measured at the end of one day of dosing at 14 days and was calculated as follows:

%TGI=(100)[1-[(the treated tumor volume _final - tumor volume _initially ) / (media treated tumor volume _final - tumor volume _initially )]]

The range of A375 and Colo ₂₀₅ represents values from two independent studies.

Example 13D: ED in Colo205 allogeneic transplantation ₅₀

Colo205 tumor cells were transplanted into male nu/nu mice. After 10 days, animals were randomized according to tumor size (range 126-256 mm ³ ) and treated with Pacific paclitaxel (IV, QOD × 5), vehicle or Compound A (PO, QD × 14) .

The pharmacokinetic parameters were obtained by administering 25 mg/kg of Compound A to Balb/c mice and extrapolating the values of the lower dose group and are shown in the table below.

Example 13E: Tumor growth inhibition rate of A375 xenograft

Compound A 50 mg/kg QD, 25 mg/kg BID, 50 mg/kg QD, and 12.5 mg/kg BID were administered to A375 allograft mice. The TGI% is calculated and plotted and shown in Figure 2.

Example 13F: Plasma concentration in mice

A375 tumor cells were transplanted into female nu/nu mice, and the cells were grown to 100-200 mm ³ . Compound A or vehicle (50 mg/kg QD, 25 mg/kg BID, 50 mg/kg QD, and 12.5 mg/kg BID) was orally administered once daily (QD) or twice daily (BID). The tumor growth inhibition rate was measured at the end of one day of dosing at 14 days and was calculated as follows:

%TGI=(100)[1-[(the treated tumor volume _final - tumor volume _initially ) / (media treated tumor volume _final - tumor volume _initially )]]

Statistical significance = logarithmic scale test

Example 13G: Inhibition of mouse xenograft tumors and brain MEK activity

A single dose of vehicle or Compound A was administered to female nu/nu mice transplanted with Colo205 tumor cells at 2.5, 5, 10 or 25 mg/kg. The amount of the compound in the plasma sample was measured and the pERK content in the tumor and brain samples was determined, which were collected at 2, 6, 12, and 24 hours after administration. The pERK content obtained by the Western blot method was quantified using LI-COR Odyssey, corrected for total ERK content and compared to the amount of vehicle treated to determine the MEK inhibition %. The MEK inhibition rate of the tumor or brain of each mouse was plotted against the corresponding plasma concentration of Compound A in the animals. Nonlinear regression analysis MEK inhibition in tumor EC ₅₀ of 73nM. The brain EC _{50 is} greater than 5000 nM.

A plot of plasma concentration (log nM) versus pERK% inhibition is shown in Figure 3.

Capsule preparation

Example 14A

A blue No. 1 hard gelatin capsule containing a dry powder blend composition of the following Compound A at a concentration of 1 mg and 10 mg (see the table shown in Example 93 above) was prepared: .

Compound A was prepared as described herein, and then a fluid energy mill (Spiral Jet Mill, electro-grinding, with a 50 mm diameter grinding chamber; 50°. 4 x 0.8 mm nozzle ring; 0.8 mm injector nozzle diameter and 3 mm) was used. The injector nozzle distance is micronized. Compound A was mixed with a portion of microcrystalline cellulose and sieved through a #20 mesh screen and added to a diffusion-roll blender (V-blender). The remaining microcrystalline cellulose was sieved through a #20 mesh screen, added to the material in the blender and blended. The croscarmellose sodium and sodium lauryl sulfate were sieved through a #20 mesh screen, added to the blender and blended. The powder blend was passed through a rotary impeller mill (Quadro CoMil) and added back to the blender and blending continued. Magnesium stearate was sieved through a #20 mesh screen and blended with the ground powder blend. This powder blend was filled into capsule No. 1. The 10 mg capsules were strip marked for identification.

The composition of the capsules is shown in the table below:

A typical batch formulation for a 10,000# batch of 1 mg capsule is as follows:

A typical batch formulation for a 10,000# batch of 10 mg capsules is as follows:

Example 14B

A blue No. 1 hard gelatin capsule containing a dry powder blend composition of the following compound A at a concentration of 1 mg and 10 mg (see the example 93 in the foregoing) was prepared. Table):

Compound A was prepared as described herein and using a fluid energy mill (Spiral Jet Mill, electro-grinding, with a 50 mm diameter grinding chamber; 50°. 4 x 0.8 mm nozzle ring; 0.8 mm injector nozzle diameter and 3 mm) The injector nozzle distance is micronized. Compound A was mixed with a portion of microcrystalline cellulose and sieved through a #20 mesh screen and added to a diffusion-roll blender (V-blender). The remaining microcrystalline cellulose was sieved through a #20 mesh screen, added to the material in the blender and blended. The croscarmellose sodium and sodium lauryl sulfate were sieved through a #20 mesh screen, added to the blender and blended. The powder blend was passed through a rotary impeller mill (Quadro CoMil) and added back to the blender and blending continued. Magnesium stearate was sieved through a #20 mesh screen and blended with the ground powder blend. This powder blend was filled into capsule No. 1. The 10 mg capsules were strip marked for identification.

The composition of the capsules is shown in the table below:

Human In vivo active

Example 15: Administration of the capsules described in Example 95A in a human cancer individual

A single dose of the 1 mg or 10 mg capsule composition described previously in Example 95A is administered to a human cancer subject. For the 2 mg dose, 2 x 1 mg capsules are administered to the individual; for the 4 mg dose, 4 x 1 mg capsules are administered to the individual; for the 6 mg dose, 6 x 1 mg capsules are administered to the individual; for the 10 mg dose, 1 x 10 mg capsules are administered to the individual; for 20 mg Dosage, 2 x 10 mg capsules are administered to the individual.

Monitor the concentration-time graph and display it in Figure 4 and in the table below:

Crystalline polymorphic form

Example 16: N-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonate Preparation of guanamine

N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide It is prepared according to the aforementioned procedure (see published international patent application WO 2007/014011) and is summarized below.

Step A :2-Fluoro-N-(2,3,5-trifluoro-6-nitrophenyl)-4-iodoaniline

A 1.0 M solution of bis(trimethyldecane)amine lithium (LiN(SiMe ₃ ) ₂ )"LHMDS" (15.37 mL, 15.37 mmol) was slowly added to the stirred 2-fluoro group at -78 ° C under nitrogen. A solution of 4-iodoaniline (3.64 g, 15.37 mmol) in dry THF (100 mL)EtOAc. 2,3,4,6-Tetrafluoronitrobenzene was added and the reaction mixture was allowed to warm to room temperature and stirring was continued for a further 16 hours. Ethyl acetate (200 mL) was added and EtOAc (EtOAc)EtOAc. MH ⁺ : 410.9. ¹ H NMR (DMSO, 300 MHz): 6.85 (t, 1 H); 7.38 (d, 1H); 7.62 (m, 2H); 8.78 (s, 1H).

Step B :2-Fluoro-N-(2,3-difluoro-5-methoxy-6-nitrophenyl)-4-iodoaniline

The stirred (2-fluoro-4-iodo-phenyl)-(2,3,5-trifluoro-6-nitro-phenyl)-amine (1.23 g, 3 mmol) The solution was cooled to -78 °C under nitrogen and a 25% solution of sodium methoxide (0.68 mL, 0.3 mmol) was slowly added. The reaction mixture was allowed to warm to room temperature and stirring was continued for a further 16 h. TLC showed that the reaction was incomplete. Ethyl acetate (100 mL) was added to EtOAc (EtOAc m. m/z = 424 [M+H] ⁺ .

Step C :5,6-difluoro-N ¹ - (2-fluoro-4-iodophenyl)-3-methoxybenzene-1,2-diamine

Ammonium chloride (1.18 g, 20.16 mmol) and iron powder (1.15 g, 21.44 mmol) were added to the stirred (2,3-difluoro-5-methoxy-6-nitro-phenyl)- ( 2-Fluoro-4-iodo-phenyl)-amine (0.57 g, 1.34 mmol) in EtOAc (20 mL). The mixture was stirred at reflux for 16 h, cooled to rt. filtered over EtOAc EtOAc. The residue was dissolved in EtOAc (EtOAc)EtOAc. MH ⁺ : 393.2. ^{1 H 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

5,6-Difluoro-N ¹ -(2-fluoro-4-iodophenyl)-3-methoxybenzene-1,2-diamine (1 eq) in anhydrous pyridine (5 ml/mmol) 1-Allyl-cyclopropanesulfonium chloride (1-5 eq) was added to the solution. The reaction mixture was stirred at 40 ° C for 48 hours. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with brine, dried (MgSO ₄ The residue was purified by flash column chromatography over EtOAc to afford title product. _{^{1 H NMR (CDCl 3, 300MHz}} ): δ7.417 (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, 1H), 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

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

Example 17: N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Preparation of hydroxypropyl)cyclopropane-1-sulfonamide

The pure S isomer was obtained by a palmitic HPLC separation of the racemic mixture. ¹ H NMR (CDCl ₃ , 300 MHz): δ 7.38 (dd, J = 1.7 & 10.3 Hz, 1H), 7.26 (m, 1H), 7.14 (s, 1H), 6.87 (s, 1H), 6. , J=6.8 & 11.4 Hz, 1H), 6.43 (m, 1H), 4.06 (m, 1H), 3.89 (s, 3H), 3.63 (dd, J = 3.7 & 11.1 Hz, 1H), 3.49 (dd, J=6.4 & 11.1 Hz, 1H), 2.3 (dd, J=9.7 & 16.1 Hz, 1H), 1.77 (dd, J=1.9 & 16.0 Hz, 1H), 1.37 (m, 1H), 1.25 (m, 1H) ), 1.21 (m, 2H), 0.86 (m, 2H); m/z = 571 [M-1] ^- .

Example 18: N-(R)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Preparation of hydroxypropyl)cyclopropane-1-sulfonamide

The pure R isomer was obtained by a palmitic HPLC separation of the racemic mixture. ¹ H NMR (CDCl ₃ , 300 MHz): δ 7.38 (dd, J = 1.7 & 10.3 Hz, 1H), 7.26 (m, 1H), 7.14 (s, 1H), 6.87 (s, 1H), 6. , J=6.8 & 11.4 Hz, 1H), 6.43 (m, 1H), 4.06 (m, 1H), 3.89 (s, 3H), 3.63 (dd, J = 3.7 & 11.1 Hz, 1H), 3.49 (dd, J=6.4 & 11.1 Hz, 1H), 2.3 (dd, J=9.7 & 16.1 Hz, 1H), 1.77 (dd, J=1.9 & 16.0 Hz, 1H), 1.37 (m, 1H), 1.25 (m, 1H) ), 1.21 (m, 2H), 0.86 (m, 2H); m/z = 571 [M-1] ^- .

Example 19: N-(S)-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Preparation of Crystalline Polymorph Form A of Hydroxypropyl)cyclopropane-1-sulfonamide

Step A

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide (216.10 g) was fed into a 4 L Erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Ethyl acetate (about 600 mL, purchased from Fisher Company) was added. Heating and stirring were initiated to form a brown suspension. The mixture was refluxed at low speed and additional ethyl acetate (ca. 200 mL) was added to effect complete dissolution to give a dark brown solution. Heptane (available from Acros Corporation) was slowly added in portions to the refluxed solution at such a rate that all precipitates formed after each addition were rapidly dissolved and maintained at reflux. After the addition of 2 L of heptane to this solution, the solid formed formed was very slowly dissolved under reflux. Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature while stirring for 16 hours. A thicker layer of crystalline material gradually forms around the surface of the glass during placement. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a 25 cm Buchner funnel equipped with Whatman #1 filter media. The collected crystals were washed with heptane (1 L) and allowed to air dry under vacuum. The crystals were further dried at 40 ° C / < 1 Torr over 20 hours to give the product as a pink crystalline solid (160.99 g, 77.2%).

Step B

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide (13.2 g) and ethyl acetate (30 mL) were fed into an Erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Stirring and heating were started to reflux at low speed to achieve complete dissolution to give a dark brown solution. The heptane is slowly added in portions to the refluxing solution at a rate such that all precipitates formed after each addition are rapidly dissolved and maintained at reflux until the addition of heptane in the solution causes the formed solid to reflux. Soluble very slowly (about 90 mL of heptane). Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature while stirring for 16 hours. A thicker layer of crystalline material gradually forms around the surface of the glass during placement. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a Buchner funnel equipped with Whitman #1 filter media. The collected crystals were washed with heptane and allowed to air dry under vacuum. The crystals were further dried at 40 ° C / < 1 Torr over 20 hours to give the product as a pink crystalline solid.

Step C

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide (44.8 g) and ethyl acetate (750 mL) were fed into an Erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Stirring and heating were started to reflux at low speed to achieve complete dissolution to give a dark brown solution. The hexane is slowly added in portions to the refluxing solution at a rate such that all precipitates formed after each addition are rapidly dissolved and maintained at reflux until the addition of hexane in the solution causes the solid formed to form under reflux. Soluble very slowly (about 2L hexane). Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature while stirring for 16 hours. A thicker layer of crystalline material gradually forms around the surface of the glass during placement. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a Buchner funnel equipped with Whitman #1 filter media. The collected crystals were washed and allowed to air dry under vacuum. The crystals were further dried at 40 ° C / < 1 Torr over 20 hours to give the product as a pink crystalline solid.

Example 20: N-(R)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Preparation of crystalline polymorph of hydroxypropyl)cyclopropane-1-sulfonamide

N-(R)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide (216.10 g) was fed into a 4 L Erlenmeyer flask equipped with a large magnetic stir bar and a magnetic stirrer/hot plate. Ethyl acetate (about 600 mL) was added. Heating and stirring were initiated to form a brown suspension. The mixture was refluxed at low speed and additional ethyl acetate (ca. 200 mL) was added to effect complete dissolution to give a dark brown solution. The heptane is slowly fed in portions to the refluxing solution at such a rate that all of the precipitate formed after each addition dissolves rapidly and maintains reflux. After the addition of 2 L of heptane to this solution, the solid formed formed was very slowly dissolved under reflux. Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature while stirring for 16 hours. A thicker layer of crystalline material gradually forms around the surface of the glass during placement. The resulting suspension was equilibrated in an ice/water bath with stirring. The suspension was filtered on a 25 cm Buchner funnel equipped with Whitman #1 filter media. The collected crystals were washed with heptane (1 L) and allowed to air dry under vacuum. These crystals were further dried at 40 ° C / < 1 Torr over 20 hours.

Example 21: IC ₅₀ Data generation

Preparation of materials and reagents: Human GST-MEK1 and the constitutively active dual gene GST-MEK1 ^CA (with mutations Ser218Asp and Ser222Asp) were sub-selected from the wild-type human MEK1 cDNA in the yeast expression vector pGEM4Z (Promega, Madison, WI). . GST-MEK1 ^CA was visualized in E. coli and partially purified using glutathione Sepharose 4B affinity resin (Amersham Pharmacia Biotech, Piscataway, NJ). The ERK2 dual gene was subcloned from the MAPK2/Erk2 cDNA (wild type) (Upstate Biotechnology, Inc., Waltham, MA) in pUSEamp in the vector pET21a (Novagen, Madison, WI) to generate an N-terminal histidine label. Mouse ERK2 dual gene. ERK2 was expressed and purified to homogeneity [Zhang, 1993 #33]. Myelin basic protein (MBP) was purchased from Gibco BRL (Rockville, MD). EasyTides adenosine 5'-triphosphate (ATP) ([γ- ³³ P]) (NEN Perkin Elmer, Wellesley, MA) is the radiolabeling source for all kinase reactions. Activated Raf-1 (truncated) and activated MAP kinase 2/ERK2 were purchased from Upstate (Lake Placid, NY). 4-20% Criterion Precast gel was purchased from Bio-Rad (Hercules, CA).

Determination of activity: The compound was diluted from a stock solution of dimethyl sulfoxide (DMSO) to 1 × HMNDE (20mM HEPES pH 7.2,1mM MgCl 2, 100mM NaCl, 1.25mM DTT, 0.2mM EDTA) in. A typical 25 ml assay contains 0.002 Nemo MEK1 ^CA , 0.02 Nemo ERK2, 0.25 Nemo MBP, 0.25 Nemo unlabeled ATP, and 0.1 μCi [γ ³³ P]ATP. The screening assay consists essentially of four additions. 5 μl of the diluted compound was dispensed into a 96-well assay plate. Then 10 μl of a 2.5× enzyme mixture (MEK1 ^CA and ERK2 only) was added to each well, followed by pre-incubation for 30 minutes at ambient temperature. Then 10 μl of a 2.5× substrate mixture (labeled and unlabeled ATP plus MBP) was added, followed by incubation at ambient temperature for 60 minutes. Finally, 100 μl of 10% trichloroacetic acid (TCA) was added and incubated at room temperature for 30 minutes to stop the reaction and precipitate the radiolabeled protein product. The reaction product was collected on 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 for 30 minutes at room temperature. A back seal was manually applied and 40 μl of scintillation cocktail was dispensed into each well. A top seal was applied and the plates were counted in a TopCount (2 sec/well). For some experiments, a truncated version of MEK that requires activation by Raf kinase was used.

Example 22: Generation of EC50 data

The effect of the compound in the cells is determined by Western blot analysis of phosphorylated ERK. MDA-MB-231 breast cancer cells were plated in 48-well plates at 20,000 cells per well and grown in a 37° humidified CO ₂ incubator. The next day, growth medium (DMEM + 10% fetal bovine serum) was removed and replaced with starvation medium (DMEM + 0.1% fetal bovine serum). The cells were cultured for 16 hours in starvation medium and then treated with a series of concentrations of compound for 30 minutes. After incubation with the compound, the cells were stimulated with 100 ng/ml EGF for 5 minutes. The cells were then lysed and analyzed by Western blotting using monoclonal antibodies raised against phosphorylated ERK. The secondary antibody was combined with a near-infrared dye to amplify the signal and detect it on a Licor Odyssey scanner. The signal intensity was quantified and this data was used to generate a dose response curve and perform an EC50 calculation.

Example 23: Activity data of compounds

The compounds described in Examples 1, 2 and 3 were tested in the above assay. The results are summarized in the table below:

Example 24: XRPD data

The XPRD was performed on an Inel XRG-3000 diffractometer equipped with a curve position sensitive detector with a 120° 2θ range. The real-time data was collected using Cu Kα radiation at a resolution of 0.03° 2θ. The tube voltage and current are set to 40kV and 30mA, respectively. A spectrum of 2.5 to 40° 2θ is presented to facilitate direct spectral comparison. By (S)-N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Hydroxypropyl)cyclopropane-1-sulfonamide (synthesized as described herein) was loaded into a thin-walled glass capillary to prepare a sample for analysis. Each capillary is moved to a motorized goniometer head to allow the capillary to rotate during data acquisition. These samples were analyzed for 5 minutes. Instrument calibration is performed daily using the 矽 reference standard. Figure 5 is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Powder x-ray diffraction (PXRD) spectrum of hydroxypropyl)cyclopropane-1-sulfonamide Form A. Figure 7 is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Powder x-ray diffraction (PXRD) spectra of hydroxypropyl)cyclopropane-1-sulfonamide Form A (upper) and amorphous (bottom).

Example 25: Differential Scanning Calorimetry (DSC)

Analysis was performed on a TA Instruments Differential Scanning Calorimeter Q1000. This instrument was calibrated using indium as a reference material. The sample was placed in a standard aluminum DSC pan with a non-crimped lid construction and its weight recorded accurately. To determine the glass transition temperature of amorphous materials (T _g), the sample tube several cycles between -40 ℃ to 140 ℃. The final temperature was raised to 150 °C. According to the inflection point of the transformation of the last cycle, T _{g is} recorded. Figure 6 is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Modulated DSC thermogram of hydroxypropyl)cyclopropane-1-sulfonamide (Form A). This graph plots the measured sample temperature (°C) with the corrected heat flux (watts/gram, W/g).

Example 26: Dynamic Vapor Sorption/Desorption (DVS)

Moisture adsorption/desorption data was collected on a VTI SGA-100 vapor adsorption analyzer. Adsorption and desorption data were collected under nitrogen nitrogen purge at a 10% relative humidity (RH) interval of 5% to 95% RH. The sample was not dried before analysis. The balance criterion for analysis is that the weight change is less than 0.0100% in 5 minutes, and if this weight criterion is not met, the maximum equilibrium time is 3 hours. The data for the initial moisture content of the sample was not corrected. Sodium chloride and polyvinylpyrrolidine were used as calibration standards. Figure 8 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di DVS isotherm of hydroxypropyl)cyclopropane-1-sulfonamide (Form A). This material exhibited negligible weight changes during the experiment.

Example 27: Thermogravimetric Analysis (TG)

Analysis was performed on a TA Instrument 2950 Thermogravimetric Analyzer. Calibration standards consisting of 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 ° C and then heated at a heating rate of 10 ° C/min under a stream of nitrogen until the final temperature was 350 ° C. Figure 9 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di TG thermogram of hydroxypropyl)cyclopropane-1-sulfonamide (Form A) showing a weight loss up to 140 ° C negligible, indicating: in polymorph, Form A is an unsolvate.

Example 28: In vitro cancer screening

Human tumor cell lines were grown in RPMI 1640 medium containing 5% fetal bovine serum and 2 mM L-glutamic acid. The cells were seeded into 96-well microtiter plates at 100 μL of plating density ranging from 5,000 to 40,000 cells/well depending on the doubling time of individual cell lines. After cell seeding, these microtiter plates were supplemented with N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl. 1-(2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide was incubated for 24 hours at 37 ° C, 5% CO ₂ , 95% air and 100% relative humidity.

After 24 hours, the two plates of each cell line were fixed in situ with TCA to express the addition of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenyl) Amino acid)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was measured for the cell population of each cell line (Tz). Making N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide was dissolved in dimethyl hydrazine in an amount 400 times the maximum test concentration to be final and stored frozen before use. Upon addition, an aliquot of the frozen concentrate was thawed and diluted to a concentration twice the maximum test concentration to be finalized with complete medium containing 50 μg/ml gentamicin. Prepare another 4 times 10 times or A logarithmic series of dilutions was provided to provide a total of 5 concentrations plus a control group. An aliquot of 100 μl of these different dilutions was added to the appropriate microtiter wells already containing 100 μl of medium to give the desired final concentration.

Addition of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxy) After propyl)cyclopropane-1-sulfonamide, the plates were incubated for 48 hours at 37 ° C, 5% CO ₂ , 95% air and 100% relative humidity. For attached cells, this assay is terminated by the addition of cold TCA. The cells were fixed in situ by slowly adding 50 μl of cold 50% (w/v) TCA (final concentration, 10% TCA) and incubated at 4 ° C for 60 minutes. The supernatant was discarded and the plates were washed 5 times with tap water and air dried. A 0.4% (w/v) solution of Sulforhodamine B (SRB) in 100% acetic acid (100 μl) was added to each well, and the plate was incubated at room temperature for 10 minutes. After dyeing, the unbound dye was removed by washing 5 times with 1% acetic acid and the plates were allowed to air dry. The bound dye was then dissolved with 10 mM Trizma base and the absorbance read at 515 nm on an automated plate reader. For suspension cells, the method was identical except that the assay was terminated by slowly adding 50 μl of 80% TCA (final concentration, 16% TCA) to fix the settled cells to the bottom of the well. 7 absorbance measurements were used [time 0 (Tz), control growth (C) and N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodobenzene) at 5 concentrations) Test growth (Ti) in the presence of arylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, at each drug concentration value Calculate the percentage of growth under. The percentage of growth inhibition is calculated as follows:

Three dose response parameters were calculated. 50% growth inhibition (GI50) was calculated based on [(Ti-Tz) / (C-Tz)] × 100 = 50, which is an increase in net protein (in SRB staining amount) in control cells during drug culture. Test) There is a 50% reduction in concentration. N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-() leading to total growth inhibition (TGI) The concentration of 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was calculated from Ti = Tz. The LC50 indicating the net loss of cells after treatment (the concentration of the drug that reduces the measured protein by 50% at the end of the drug treatment compared to the start) is based on [(Ti-Tz)/Tz] × 100 = -50 Calculation. If the activity value is reached, the value of each of the three parameters is calculated; however, if the effect is not reached or exceeded, the value of the parameter is expressed as greater or less than the maximum or minimum concentration tested.

Discussion groups corresponding to leukemia, non-small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer were examined for the indicated cell lines, and the results are shown in the following table.

Example 29: In vitro Antiproliferative activity

In this example, the test for N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, The following effects of 3-dihydroxypropyl)cyclopropane-1-sulfonamide: (1) activity against the growth of several tumor cell lines with different mutations (GI ₅₀ ); (2) for several B-Raf The growth activity of the mutant cell line (GI ₅₀ ); (3) the effect on the growth of fixation-independent cells; (4) the effect on the cell cycle; and (5) the toxic effects on the primary liver and kidney cells.

Example 29A: Cell Culture/Growth Inhibition Assay

Human melanoma A375 cells and human colon cancer Colo205 cell lines were obtained from ATCC (Manassas, VA). A375 cells were maintained in DMEM supplemented with 10% fetal calf serum, glutamic acid (2 mM), penicillin (100 U/ml) and streptomycin (100 μg/ml). The cells were maintained at 37 ° C, 5% CO ₂ and 100% humidity. Colo205 cells were maintained in RPMI supplemented with 10% fetal calf serum, glutamic acid (2 mM), penicillin (100 U/ml), and streptomycin (100 μg/ml). For growth inhibition experiments, cells were plated at 1000 cells/20 μl/well in white 384-well microplates. After 24 hours, 5 μl of 5X drug stock solution was added. All drugs were initially prepared as a 200x stock solution in DMSO, so the final DMSO concentration was 0.5%. The cells were incubated at 37 ° C for 48 hours and the ATP content was determined using CellTiterGlo (Promega, Madison, WI). Adenylate kinase (AK) release was determined using Toxilight (Cambrex, Walkersville, MD). Non-linear curve fitting was performed using GraphPad Prism 4 (GraphPad Software, San Diego, CA). 4-amino-8-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-pyridinium [2,3-d]pyrimidine- 5(8H)-ketone (VRX-14686) is a cytotoxic agent used as a reference compound.

Growth inhibition (%) = (media-only control value (RLU)-N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxy Phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide RLU)/(media-only control RLU-1μM N-(S)-(3,4-difluoro- 2-(2-Fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide RLU); The ATP content is determined by N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) , 3-dihydroxypropyl)cyclopropane-1-sulfonamide is based on growth arrest.

Cell survival (%) = (N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) , 3-dihydroxypropyl)cyclopropane-1-sulfonamide RLU-10μM VRX-14686 RLU)/(media-only control RLU-10μM Tamoxifen RLU); when measuring ATP content Based on cell killing caused by VRX-14686.

Cell killing (%) = (N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide RLU-only vehicle control RLU)/(10 μM tamoxifen RLU-media-only control RLU); when measuring ATP content Based on the killing of cells caused by tamoxifen.

RLU = relative illuminating unit

Example 29B: Assessment of cell cycle arrest

A375 cells were plated at 10,000 cells/200 μl/well in 96-well microplates. After 24 hours, the cells approached 50% confluence and 50 μl of the 5X drug stock solution was added. After a further 24 hours, the cells were trypsinized, fixed in 200 μl Prefer (Anatech, Battle Creek, MI) and stored at 4 ° C overnight. The cells were then washed in PBS, permeabilized and stained in 0.1% Triton X-100, 200 μg/ml DNase-free RNase and 25 μg/ml propidium iodide (Molecular Probes, Sunnyvale, CA), and in Guava PCA Analysis was performed on a -96 (Guava Technologies, Foster City, CA). Data was analyzed using ModFit LT (version 3.0, Verity, Topsham, ME).

Evaluation of fixation-independent cell growth inhibition

The wells of the "ultra-low binding" plate (Corning, Acton MA) were filled with 60 μl of a 0.15% agarose solution in complete RPMI. Next, 60 μl of complete RPMI containing 9,000 Colo205 cells in 0.15% agarose was added to each well. After 24 hours, 60 μl of 3X drug solution in complete RPMI without agarose was added. After 7 days, 36 μl of 6X MTS reagent (CellTiter 96 Aqueous, Promega, Madison, WI) was added to each well. After 2 hours at 37 ° C, the absorbance at 490 nm was measured on an M5 plate reader (Molecular Devices, Sunnyvale, CA). Non-linear curve fitting was performed using GraphPad Prism 4.

Growth inhibition against MEK-dependent cancer cell growth (GI ₅₀ )

B-Raf mutant cells A175 (human melanoma), A431 (melanoma), Colo205 (colon cancer), HT29 (colorectal adenocarcinoma), MDA-MB231 (breast cancer), and BxPC3 (pancreas) Cancer) exposure to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3- Dihydroxypropyl)cyclopropane-1-sulfonamide was analyzed for 48 hours and analyzed for ATP content. Using 1 μM-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl) The cyclopropane-1-sulfonamide was assayed for 100% growth arrest.

The following table shows the cell lines from each of at least three experiments The average GI value of _50, and the display N- (S) - (3,4- difluoro-2- (2-fluoro-4-iodo-phenylamino) -6-Methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide in three B-Raf mutant cell lines (A375, Colo205 and HT29) and a ras/ Growth inhibition was induced in the raf/MEK/MAPK pathway wild-type cell line (A431) with an average potency of 79 nM (±9 nM).

In an independent study, the log-phase split B-Raf mutant cells A375 (human melanoma), SK Mel 28 (human melanoma), and Colo205 (human colon cancer) were exposed to N-(S)-( 3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonate The indoleamine was taken for 48 hours and analyzed for ATP content. The following table shows the GI _{50 of} each cell line indicating N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl ) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide Amides cause performance close growth inhibition ₅₀ values of inhibiting MEK EC thereof.

Figures 10A and 10B show exposure to increasing concentrations of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) The growth of the logarithmicly dividing A375 cells of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was stagnant. Analyze the ATP content of the cells. Using 1 μM N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxyl Propyl)cyclopropane-1-sulfonamide was assayed for 100% growth arrest.

Cytotoxic lysis of cell supernatants was analyzed by measuring the release of adenylate kinase (AK). Exposure of log phase split A375 cells to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- (2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide and PD-325901 lasted 48 hours. (100% cell killing was determined using 20 μM tamoxifen). The results are shown in Figure 11 . This data indicates N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl in several sensitive human cancer cell lines. --1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide causes non-toxic growth arrest by (i) growth arrest (ATP quantification); and (ii) lack of cytotoxicity Cytolysis (AK release) is demonstrated. Lack of AK release was confirmed for all cell lines tested.

Example 29C: Fixation independent growth inhibition

Quantitative assessment of exposure to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1 in a 96-well microplate format -(2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide 7 days of fixation independent growth of Colo205, A375 and MDA-MB231 cells. Viability was determined by MTS assay. The GI ₅₀ values are shown below:

Figures 12A-12C show N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3 -Dihydroxypropyl)cyclopropane-1-sulfonamide against (A) human colorectal cancer Colo205 cells (GI ₅₀ =11 nM), (B) A375 cells (GI ₅₀ =22 nM) growth inhibition and (C) Inhibition of MDA-MB231 cells, which did not show N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)- in a two-dimensional fixation-dependent assay 6-Methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide causes growth arrest.

Exposure of log phase split A375 cells to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- (2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide (1 μM) was analyzed for growth inhibition (ATP content) and cytotoxic lysis (AK release) of the cell supernatant over 48 hours. 100% survival (ATP assay) was determined in vehicle-only control wells. The results shown in the table below indicate 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.

Example 29D: Fixation independent growth inhibition

Quantitative independent growth was assessed quantitatively in a 96-well microplate format. Figure 13A shows inhibition of growth of human Colo205 cells of colorectal cancer and ₅₀ and the value of 6nM under the GI 11nM respectively. Figure 13B shows inhibition of growth of A375 cells and the GI ₅₀ values of 5nM and 22nM in at.

Example 29E: N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Cell cycle analysis of growth arrest caused by hydroxypropyl)cyclopropane-1-sulfonamide

MEK inhibition has been shown to cause G1/S phase cell cycle arrest in A375 cells.

Exposure of log phase split A375 cells to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- (2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide The percentage of cells stained for a phase dependent amount of intracellular DNA was determined using flow cytometry for 24 hours.

The following table shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3- Dihydroxypropyl)cyclopropane-1-sulfonamide and control (% of cell distribution in the respective growth phase of vehicle-only cells).

14A and 14B show N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, Effect of 3-dihydroxypropyl)cyclopropane-1-sulfonamide on cell cycle progression, showing exposure of A375 cells to N-(S)-(3,4-difluoro-2-(2-fluoro-4) -iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide causes stagnation of the G1 phase of the cell cycle, which is caused by G2 And the cell consumption at the time of S phase is indicated.

Example 29F: Assessment of primary hepatocyte and renal cytotoxicity

Refrigerated rat liver cell lines were obtained from CellzDirect (Austin, TX) and applied to collagen coated 96-well plates according to the manufacturer's instructions. Drugs were added 4 hours after application to the plate (final DMSO concentration 0.5%). The coated human hepatocyte cell line was obtained from Cellz Direct and processed according to the manufacturer's instructions.

Refrigerated human renal proximal tubular epithelial cells (RPTEC) were obtained from Cambrex and processed according to the manufacturer's instructions. Cells were expanded for 4 days and then plated onto 96-well plates at 50,000 cells/well for drug exposure.

After 48 hours, the supernatant AK content was determined using Toxilight and the cell ATP content was determined using CellTiterGlo. The complete lethal value was determined using 15 μM VRX-14686.

The results are shown below. Very little cell lysis was observed. 30μM N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxybenzene in newly implanted primary human hepatocytes Minimal toxicity (81% survival) was observed under keto-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. RPTEC cells showed dose-dependent ATP depletion and significant cytolysis (30 μM).

The above data indicates 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 of specific human cancer cells in a fixation-dependent proliferation assay (GI ₅₀ values in the range of 70-89 nM), but does not result in Such as toxicity as determined by cell lysis assay; (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 specific human cancer cells in a fixation-dependent and independent proliferation assay with GI ₅₀ values of 51 nM, respectively And 22 nM; (3) N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, 3-dihydroxypropyl)cyclopropane-1-sulfonamide causes G1 arrest in A375 cells and inhibits fixation-independent growth, which provides evidence of anticancer activity in a physiologically relevant in vitro model; 4) N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxyl) Propyl)cyclopropane-1-sulfonamide is shown to target primary normal human hepatocytes, human renal proximal tubular epithelial cells, and Rat hepatocytes almost no cytotoxicity.

Example 30: N-(S)-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Pharmacokinetic profile of hydroxypropyl)cyclopropane-1-sulfonamide in cancer individuals after multiple doses

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl is administered multiple times at 2, 4 or 6 mg per individual N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenyl) after 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide Amino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide is readily absorbed by an average _Tmax ranging from 1.33 to 1.50 hours . The average _Cmax , C[ _tau], and AUC values increase with dose in a manner proportional to the dose. Separately, the cumulative index range is between 1.49 and 1.76 for _Cmax and between 1.90 and 2.07 for AUC, indicating a modest accumulation. Although the half-life cannot be accurately measured after multiple administrations due to limited sampling time, it is expected that the cumulative index half-life may be longer than 22 hours after multiple administrations. These half-life values are significantly longer than the half-life observed in the mouse efficiency model, which is observed to be within the typical range of 2-3 hours. In addition, the ideal peak to valley ratio is seen in all doses.

Example 31: N-(S)-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Pharmacokinetic profile of hydroxypropyl)cyclopropane-1-sulfonamide in healthy volunteers after multiple doses

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)- is administered multiple times at 10 or 20 mg per individual. N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino) after 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide 6-Methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide is readily absorbed in an average _Tmax ranging from 2.00 to 2.25 hours. The average _Cmax , C[ _tau], and AUC values increase with dose. Separately, the cumulative index range is between 1.14 and 1.23 for _Cmax and between 1.24 and 1.29 for AUC, indicating a slight accumulation. For both dose regimes, the half-life is similar and ranges from 13 to 15 hours. These half-life values are shorter than those observed in cancer individuals.

Example 32: In vitro Antiproliferative activity

Examination of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyl in a cell line derived from human gastric cancer in a cell proliferation assay Effect of phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on inhibition of cell proliferation.

Example 32A: Cell Culture/Growth Inhibition Assay:

The human gastric cancer Hs746t cell line was obtained from ATCC (Manassas, VA). Hs746t cells were maintained in DMEM supplemented with 10% fetal bovine serum, penicillin (100 U/ml) and streptomycin (100 μg/ml). The cells were maintained at 37 ° C, 5% CO ₂ and 100% humidity. For cell proliferation experiments, cells were plated at 3000 cells/100 μl/well in white 96-well plates with clear substrates. After 24 hours, the cell culture medium was removed and containing different doses of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxy Medium replacement of phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. After incubation for 48 hours at 37 ° C, ATP content was determined using CellTiterGlo (Promega, Madison, WI) and luminescence values were read using LJL Biosystems Analyst HT (Sunnyvale, CA). The ATP content for each dose was determined in triplicate using separate wells.

Relative cell number = (average RLU(N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( 2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide treatment)) / (average RLU of vehicle-only control).

Figure 19 shows the number of cells (relative to vehicle) versus N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) A plot of the concentration of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide and showing N-(S)-(3,4-difluoro-2-(2-fluoro-4-) Iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide inhibited proliferation of human gastric cancer Hs746t cells after 48 hours of treatment.

Example 33: In vitro Antiproliferative activity

Examination of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyl in a cell line derived from human gastric cancer in a cell proliferation assay Effect of phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on inhibition of cell proliferation.

Example 33A: Cell Culture/Growth Inhibition Assay

The human gastric cancer AGS cell line was obtained from ATCC (Manassas, VA). AGS cells were maintained in DMEM/F12 supplemented with 10% fetal calf serum, penicillin (100 U/ml) and streptomycin (100 μg/ml). The cells were maintained at 37 ° C, 5% CO ₂ and 100% humidity. For cell proliferation experiments, cells were plated at 3000 cells/100 μl/well in white 96-well plates with clear substrates. After 24 hours, the cell culture medium was removed and containing different doses of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxy Medium replacement of phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. After 3 days of incubation at 37 ° C, ATP content was determined using Cell Titer Glo (Promega, Madison, WI) and luminescence values were read using LJL Biosystems Analyst HT (Sunnyvale, CA). The ATP content for each dose was determined in triplicate using separate wells. In another experiment, cells were plated at 1000 cells/100 μl/well and cells were treated for 6 days and assayed as described above.

Relative cell number = (average RLU(N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( 2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide treatment)) / (average RLU of vehicle-only control).

15A and 15B show exposure to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide (A) for 3 days and (B) after 6 days N-(S)-(3,4-difluoro-2-(2-fluoro) -4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide concentration on cell number (relative to vehicle) Graph showing 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 AGS cell lines.

Example 34: Different amounts of N-(S) - (3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) Growth reaction of in situ human Hep3B tumor in nude mice treated with ,3-dihydroxypropyl)cyclopropane-1-sulfonamide

Evaluation of N-(S)-(3,4-difluoro-2-(2-fluoro-4-) in BALB/c nu / nu mice compared to the optimal dose of 5-fluorouracil (75 mg/kg) Iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide ("Compound A") inhibits in situ Hep3B2.1- 7 dose response efficacy in the development of human liver cancer.

Animals :

Female BALB/c nu / nu mice (University of Adelaide, Waite Campus, SA, Australia) were used in this study, 10-14 weeks old, and the body weight range was as follows: 19.1-29.94 g (average 22.95 g). The mice were divided into 6 study groups (4 treatment groups and 2 control groups) as follows: number of mice per group: 10 in groups 1 to 5; in 'acceptance rate (Take-Rate) '15 in the control group (Group 6) maintained the mice in a controlled environment with a 12-hour light/12-hour dark cycle under shielded (isolated) conditions (target range: temperature 21 ± 3 ° C, humidity 30 -70%, air exchange 10-15 times per hour). Continuously monitor temperature and relative humidity. Animals were given ad libitum access to a commercially available rodent diet (Rat and Mouse Cubes, Specialty Feeds Pty Ltd, Glen Forrest, Western Australia) and tap water. All food and water supplies are sterilized using autoclave treatment.

Tumor inoculation :

Hep3B human hepatoma cells (second passage from working stock VP-Stock 353) were cultured in RPMI 1640 cell culture medium supplemented with 10% FBS and penicillin-streptomycin (50 IU/mL final concentration). Cells were harvested via trypsinization, washed twice in HBSS and counted. The cells were then resuspended in HBSS: Matrigel (1:1, v/v) and adjusted to a final volume containing 1 x 10 ⁸ cells/mL. Prior to inoculation, the incision site was adequately scrubbed with alcohol and cut through the abdominal wall to expose the liver. 10 μL of cells (1 × 10 ⁶ cells) were released by introducing the needle through the surface of the liver. Hold the needle in this position for about 30 seconds to allow Polymerize to avoid leakage of tumor cells into the abdominal cavity.

Treatment was started 14 days after inoculation. On day 7 of the study (21 days after inoculation), all mice in the 'acceptance rate' control group were sacrificed and the presence of tumors in the liver was visually assessed.

Material :

The following materials were obtained from their respective suppliers.

A sterile saline solution (0.9% NaCl (aq)) was obtained from Baxter Healthcare Australia, Old Toongabbie, NSW, Australia. Cetyl polyoxyethylene ether EL was obtained from Sigma-Aldrich Pty Ltd, Castle Hill, NSW, Australia. 5-fluorouracil (clinical formulation, clear, colorless liquid) was obtained from Mayne Pharma Pty Ltd. RPMI 1640 cell culture medium, FBS and HBSS were obtained from Invitrogen Australia 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 cell line is derived from the American Type Culture Collection (ATCC), Rockville, MD, USA. Matrigel The lines were obtained from BD Biosciences, North Ryde, NSW, Australia.

Use in inoculation suspension It can improve the acceptance rate of tumors and reduce the variability of tumor size, and the growth of Hep3B2.1-7 human liver cancer is stable when inoculated in the presence of this extracellular matrix.

Preparation and administration of compounds :

Cetyl polyoxyethylene ether EL: brine (1:9, v/v; vehicle control), N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodobenzene) Aminoamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide ("Compound A") or 5-fluorouracil (Compound Control) According to the following time schedule:

The vehicle control cetyl polyoxyethylene ether EL: saline (1:9, v/v) was orally administered once a day for 21 days (days 0 to 20) in a dose volume of 10 mL/kg.

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide was formulated in cetyl polyoxyethylene ether EL: brine (1:9, v/v). Stock solutions were prepared once a week and stored at 4 °C. The dosing solutions were prepared on the day of each administration. N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl The cyclopropane-1-sulfonamide was orally administered once a day for 21 days (days 0 to 20) in a dose volume of 10 mL/kg. This compound was 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 75 mg/kg at a dose of 10 mL/kg once a week for 3 weeks (on days 0, 7 and 14) ).

No treatment was applied to the mice in the sixth group ("acceptance rate" control group). On the 7th day of the study (21 days after inoculation), the mice were sacrificed and the liver was exposed to determine the "acceptance rate" and size of the tumor in the liver wall.

The body weight of each animal was measured immediately before administration. The volume administered to each mouse was calculated and adjusted based on body weight.

Tumor measurement:

When the liver and tumor were excised after the death of the mice on the day of the study, the liver and tumor wet weight were measured. At the end of the study, livers were harvested from all mice in each study group and weighed. If there is a visible tumor, the number of visible tumors is counted. These tumors were removed from the liver and weighed.

Data measurement and sample collection time history

Data acquisition and calculation:

Sensors for each animal (Bar Code Data Systems Pty Ltd, Botany Bay, NSW) were scanned using a barcode reader (LabMax I, DataMars, Switzerland) before data was acquired. All measurements were taken with the same hand caliper (Absolute Digimatic Model CD-6" CS, Mitutoyo Corporation, Japan). Use Pendragon Forms 4.0 ( Software Corporation, Libertyville, IL, USA) as a transport software synchronizes data with vivoPharm's security-relevant database. Use AIDAM v2.4 for data reporting and data calculation.

Statistics and calculations:

All statistics and calculations were performed using SigmaStat 3.0 (SPSS Australasia Pty Ltd, North Sydney, NSW, Australia).

A double sample t test was used to determine the significance of body weight changes between day 0 and day of end of the study in a treatment group. A Mann-Whitney rating and test is performed when the data fails the normality test or the equal difference test.

Kruskal-Wallis One-Way Analysis of Variance (Kruskal-Wallis One-Way Analysis of Variance (ANoVA) on Ranks). The same statistical analysis was performed on the data of mice with tumors in the study.

A p value of less than 0.05 was considered significant.

Liver weight and tumor weight data of tumor-bearing mice and average weight of liver and tumor of each mouse in each group of tumor-bearing mice

A sample of mice of Group 5 (75 mg/kg 5-fluorouracil) was not collected and this group was killed during the study. Since there were large tumors in some mice, as indicated by the appearance of the swollen abdomen, this study was terminated 18 days after the initial treatment.

N-(S)-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The dose-dependent reduction trend of liver and tumor weight in the cyclopropane-1-sulfonamide treatment group was significant. When considering only mice with tumors, the highest dose of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxy was found. Phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide (Group 4, 50 mg/kg) and 5-fluorouracil (Group 5, 50 mg/kg) treated group The mean liver weight was significantly different from the vehicle control group (Group 1; p < 0.05). And found that N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3- Tumors of the group treated with dihydroxypropyl)cyclopropane-1-sulfonamide (Groups 3 and 4, respectively, 10 mg/kg and 50 mg/kg) and 5-fluorouracil (Group 5, 75 mg/kg) The average weight was significantly different from the vehicle control group.

These results are presented graphically in Figure 16 (mean liver weight - tumor-only mice) and Figure 17 (liver tumor weight - tumor-only mice).

Example 35: Different amounts of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) Growth reaction of in situ human HT-29 colon tumor in nude mice treated with 3-hydroxypropyl)cyclopropane-1-sulfonamide

Evaluation of N-(S)-(3,4-difluoro-2-(2-fluoro-4-) in BALB/c nu/nu mice compared to the optimal dose of 5-fluorouracil (75 mg/kg) Iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide ("Compound A") inhibits in situ HT-29 human Dose response efficacy in the development of colorectal adenocarcinoma.

Animals :

Female BALB/c nu / nu mice (University of Adelaide, Waite Campus, SA, Australia) were used in this study, 7-12 weeks old, and the body weight range was as follows: 16.58-25.39 g (mean 21.52 g). The mice were divided into 6 study groups (4 treatment groups and 2 control groups) as follows: number of mice per group: 10 in groups 1 to 5; in the 'acceptance rate' control group Nine (Group 6) maintained the mice in a controlled environment with a 12-hour light/12-hour dark cycle under barrier (isolation) conditions (target range: temperature 21 ± 3 ° C, humidity 30-70%, hourly Air exchange 10-15 times). Continuously monitor temperature and relative humidity. Animals were given ad libitum access to a commercially available rodent diet (Rat and Mouse Cubes, Specialty Feeds Pty Ltd, Glen Forrest, Western Australia) and tap water. All food and water supplies are sterilized using autoclave treatment.

Tumor inoculation :

HT-29 human colorectal adenocarcinoma cells (from passage 4 of working stock VP-Stock 325) were cultured in RPMI 1640 cell culture medium supplemented with 10% FBS and penicillin-streptomycin (50 IU/mL final concentration). Cells were harvested via trypsinization, washed twice in HBSS and counted. The cells were then resuspended in HBSS and adjusted to a final volume containing 2 x 10 ⁸ cells/mL. Prior to inoculation, the incision site was adequately scrubbed with alcohol and cut through the abdominal wall to expose the cecal wall. 5 μL of cells (1 × 10 ⁶ cells) were released by introducing the needle through the surface of the cecum wall.

material :

The following materials were obtained from their respective suppliers.

A sterile saline solution (0.9% NaCl (aq)) was obtained from Baxter Healthcare Australia, Old Toongabbie, NSW, Australia. Cetyl polyoxyethylene ether EL was obtained from Sigma-Aldrich Pty Ltd, Castle Hill, NSW, Australia. 5-Fluorouracil (clinical formulation, clear, colorless liquid) was obtained from Mayne Pharma Pty. RPMI 1640 cell culture medium, FBS and HBSS were obtained from Invitrogen Australia Pty Ltd, Mt Waverley, VIC, Australia. Penicillin-streptomycin and trypan blue are obtained from Sigma-Aldrich, Castle Hill, NSW, Australia. The HT-29 human colorectal adenocarcinoma cell line is derived from the American Type Culture Collection (ATCC), Rockville, MD, USA.

Preparation and administration of compounds :

Cetyl polyoxyethylene ether EL: brine (1:9, v/v; vehicle control), N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodobenzene) Aminoamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide or 5-fluorouracil (control compound) was administered according to the following schedule :

The vehicle control cetyl polyoxyethylene ether EL: saline (1:9, v/v) was orally administered once a day for 21 days (days 0 to 20) in a dose volume of 10 mL/kg.

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane The cyclopropane-1-sulfonamide was formulated in cetyl polyoxyethylene ether EL: brine (1:9, v/v). Stock solutions were prepared once a week and stored at 4 °C. The dosing solutions were prepared on the day of each administration. N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl The cyclopropane-1-sulfonamide was orally administered once a day at a dose of 2, 10, and 50 mg/kg in a dose volume of 10 mL/kg for 21 days (days 0 to 20).

The 5-fluorouracil clinical formulation was diluted in sterile saline and administered intravenously via the tail vein at a concentration of 75 mg/kg at a dose of 10 mL/kg once a week for 3 weeks (on days 0, 7 and 14) ).

No treatment was applied to the mice in the sixth group ("acceptance rate" control group). On the 7th day of the study (21 days after inoculation), the mice were sacrificed and the colon was exposed to determine the "acceptance" and size of the tumor in the cecal wall.

The body weight of each animal was measured immediately before administration. The volume administered to each mouse was calculated and adjusted based on body weight.

Tumor measurement:

When the cecum and tumor were harvested after the death of the mice on the day of the study, the cecum and tumor wet weight were measured. At the end of the study, all mice from each study group were culled and weighed while the tumor remained intact. Tumors were then removed from the cecum and weighed.

At the end of the study, livers were also excised from all mice in each group and fixed in 10% buffered formalin. Five liver samples from the vehicle control group were embedded in paraffin, sectioned and stained with hematoxylin-eosin stain (H&E) for histological evaluation of morphological changes.

Data measurement and sample collection time history

Data acquisition and calculation:

Sensors for each animal (Bar Code Data Systems Pty Ltd, Botany Bay, NSW) were scanned using a barcode reader (LabMax I, DataMars, Switzerland) before data was acquired. All measurements were taken with the same hand caliper (Absolute Digimatic Model CD-6" CS, Mitutoyo Corporation, Japan). Use Pendragon Forms 4.0 (Pendragon Software Corporation, Libertyville, IL, USA) as a transport software synchronizes data with vivoPharm's security-relevant database. Use AIDAM v2.4 for data reporting and data calculation.

Statistics and calculations:

All statistical calculations were performed using SigmaStat 3.0 (SPSS Australasia Pty Ltd, North Sydney, NSW, Australia).

The double sample t test was used to determine the significance of body weight change between day 0 and end of day for this study in a treatment group. N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2) at 2 and 50 mg/kg In the group treated with 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, the treatment was interrupted early due to excessive weight loss. In these groups, a double sample t test was used to determine the significance of body weight change between day 0 and the end of the study in the treatment group and between the last treatment day and the end day of the study in a treatment group. A Mann-Whitney rating and test is performed when the data fails the normality test or the equal difference test.

Single factor variability analysis (ANOVA) was performed at the end of the study on cecal weight and tumor weight data (all pairwise multiple comparison programs and multiple comparisons versus control). When the data fails the normality test, convert the value to a natural logarithm before proceeding with the program.

A p value of less than 0.05 was considered significant.

Observations:

Mean body weight loss was measured in all study groups (including vehicle control). Diarrhea and dehydration symptoms (impaired skin elasticity) were observed in all study groups (including the vehicle control group). Severe weight loss during the study period resulted in the lowest dose (2 mg/kg) and the highest dose (50 mg/kg) of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodine) Treatment of the group of phenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on the 9th and 7th day of the study, respectively Stop when. Because it accepts 10 mg/kg of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, The weight loss of the group of 3-dihydroxypropyl)cyclopropane-1-sulfonamide was less severe, so all treatments in this group were administered as scheduled. The mean weight loss at the end of the study in this group and the 5-fluorouracil treatment group was significant.

Although the acceptance rate of HT-29 tumors in the 'acceptance rate' group was 100% 21 days after inoculation, the size of these tumors was far less than expected. This may be due to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3) There was no significant difference in mean cecum and tumor weight between the -dihydroxypropyl)cyclopropane-1-sulfonamide treated group and the vehicle control group. 5-fluorouracil also had no effect on cecal and HT-29 tumor weight.

body Weight measurement (±SEM) (last day of treatment and study termination date)

Body weight data for Group 6 ("Acceptance" control group) was not collected. On day 7 of the study (21 days after inoculation), the group was sacrificed to visually assess whether tumor growth was appropriate for the purposes of this study.

Group 2 (2 mg/kg of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( Treatment of 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide) was discontinued on study day 9 and group 4 (50 mg/kg of N-(S)-(3,4-difluoro-2) -(2-Fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide) 7 days of interruption, this is due to excessive weight loss in mice. The remaining groups received all scheduled treatments during the study period.

The mean tumor weight for each group is shown in Figure 18 . The average tumor weight for each group included only survivors until the last day of the study. The values of mice that died during the study were not included in the calculated mean.

Cecal weight and tumor weight data

The shaded boxes indicate samples collected from mice that died during the study. This value is excluded from the calculation of the mean value of cecal weight and tumor weight. The trend is shown at 10 mg/kg of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( HT-29 tumor and cecal weight data were reduced after treatment with 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide.

Example 36: Delay in tumor growth in nude mice bearing human A375 melanoma xenografts

Six groups (n=9) of tumorized mice were used. The control group consisted of receiving 10% hexadecanol ethoxylate EL/saline vehicle by oral gastric tube (po) once daily for 14 days (qd×14), and intravenous injection of 30 mg/kg (iv) The second specific paclitaxel was used as a reference agent, administered one group (qod x 5) five times every other day. Four experimental groups received 25 mg/kg or 50 mg/kg, qd×14, or 12.5 or 25 mg/kg, bid×14 orally N-(S)-(3,4-difluoro-2-(2-fluoro-) 4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide ("Compound A"). Treatment results were assessed by TGD, which was defined as the difference in tumor volume between the treatment group and the control group at the median endpoint time. Toxicity was assessed by body weight measurement and clinical observation.

animal:

Female athymic nude mice ( nu / nu , Harlan) were 10 to 11 weeks old and had a body weight (BW) range of 19.3 to 25.5 grams on day 1 of the study. Animal free access to water (reverse osmosis, 1ppm Cl) and NIH31 modification and irradiation Lab It consists of 18.0% crude protein, 5.0% crude fat and 5.0% crude fiber. The mouse is housed in a static micro-separation chamber The laboratory animal pad is in a 12 hour photoperiod with 21-22 ° C (70-72 ° F) and 40-60% humidity. Adhere to the recommendations for restrictions, grazing, surgical procedures, feeding and fluid regulations, and veterinary care for laboratory animal feeding management and use practices.

Tumor transplantation:

Allogeneic transplantation was initiated by A375 human melanoma by continuous transplantation in athymic nude mice. A375 tumor fragments (~1 mm ³ ) were subcutaneously transplanted into the right abdomen of each test mouse, and the average size of tumor growth was monitored to be 100-150 mm ³ . After 13 days, the day designated as the day of the study, the animals were divided into 6 groups and each group consisted of 9 mice (reduced from 10) with individual tumor volumes ranging from 63 to 221 mm ³ and each The mean tumor volume of the group was 125.3 to 125.9 mm ³ . Tumor volume was calculated using the following equation: tumor volume (mm ³ ) = (w ² × l) / 2 , where w = width of the A375 tumor (mm) and l = length of the A375 tumor (mm).

material:

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl The cyclopropane-1-sulfonamide was dissolved in a 10% aqueous solution of 10% hexadecanol polyoxyethylene ether EL at 5 mg/ml, during which ultrasonic treatment, shaking and heating were carried out to 35 ° C to aid dissolution. A 5 mg/mL solution was used as a treatment administration solution for 50 mg/kg, and a treatment administration solution of 25 mg/kg and 12.5 mg/kg was prepared by a serial dilution method. The dosing solution was stored at room temperature for one week without exposure to light.

The paclitaxel (NPI) dosing solution was prepared from a 30 mg/mL stock solution by diluting to 3 mg/mL in 5% ethanol, 5% cetyl polyoxyethylene ether EL in 5% dextrose in water (D5W). Use every day. The paclitaxel dose was 30 mg/kg.

treatment :

The table below shows the treatment plan.

Group 1 mice received a vehicle consisting of a 10% aqueous solution of hexadecanol polyoxyethylene ether EL, administered orally by gastric tube (po), administered 14 times a day (qd x 14), and used as a tumor. Progressive control group. Group 2 animals were administered intravenously (iv) with 30 mg/kg of paclitaxel as a reference, administered five times a day (qod x 5). Mice in groups 3-6 received N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyl orally in the following schedules, respectively. Phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide: 50 mg/kg, qd×14; 25 mg/kg, twice daily for 14 days, on the first and last day A single dose (bid x 14); 25 mg/kg, qd x 14; and 12.5 mg/kg, bid x 14 were administered. All doses were administered in a volume of 0.2 mL per 20 g body weight, the scale being in accordance with the animal's body weight.

end:

Tumors in all groups were measured twice a week using calipers. Each animal was euthanized when the tumor reached a site size of 2000 mm ³ or when either of the last day of the study (Day 60) arrived first. The endpoint time (TTE) of each mouse was calculated by the following equation: TTE (days) = (log ₁₀ (endpoint volume, mm ³ )-b)/m, where b is the intercept and m is the log-transformed tumor growth data The slope of the line obtained by linear regression of the group.

This data set contains the first observation that exceeds the study endpoint volume and three consecutive observations immediately before the endpoint volume is reached. Animals that did not reach the end point specified a TTE value equal to the last day of the study. Animals classified as having an accidental NTR (non-therapeutic correlation) (NTRa) or died for unknown reasons (NTRu) were excluded from the TTE calculation (and all further analyses). Animals classified as TR (treatment-related) death or NTRm (non-therapeutic correlation due to metastasis) were assigned a TTE value equal to the day of the study death.

Treatment results were determined by tumor growth delay (TGD), which was defined as the increase in endpoint time (TTE) of the treatment group compared to the control group: TGD = TC, expressed in days, or in the middle of the TTE of the control group. The percentage indicates: Where: T = intermediate TTE of the treatment group, C = intermediate value of TTE of the control group (Group 1).

Treatment may result in partial regression (PR) or complete regression (CR) of the animal's tumor. PR in the reaction, during the course of the study, tumor volume for three consecutive measurements on day 1 of a 50% by volume or less, and equal to or greater than 13.5mm ³ In this one or more of three measurements . In the CR reaction, tumor volume was less than 13.5 mm ³ in three consecutive measurements during the course of the study. At the termination of the study, animals with a CR response were additionally classified as tumor free survivors (TFS). Tumor regression was monitored and recorded.

side effect:

Animals were weighed daily for the first five days of the study and then weighed twice a week. Obvious symptoms of any adverse, treatment-related side effects of this mouse were frequently observed, and clinical symptoms at the time of observation were recorded. Acceptable tolerance is defined as a group of treatment-related deaths with a mean weight loss of less than 20% during the test and no more than one in the animal group. Any dosing regimen that does not meet these criteria is considered to be greater than the maximum tolerated dose (MTD). Death is classified as TR if it is confirmed by medical symptoms and/or anatomy as a side effect of treatment, or is classified as TR if it dies due to an unknown cause during administration or during the last 10 days of administration. Death is classified as NTR if there is no evidence that death is associated with treatment side effects.

Statistical and graphical analysis:

Logarithmic scale tests were used to analyze the significance of the difference in TTE values between the treated and control groups. Two-tailed statistical analysis was performed at a significant amount of P = 0.05.

The median value of the tumor growth curve shows the median tumor volume of each group as a function of time. When the animals withdrew from the study due to tumor size or TR death, the data for the intermediate tumor volume calculated for the subsequent time points included the final tumor volume recorded for the animals. This curve was truncated after 50% of the animals in the group developed the tumor and withdrew from the study. Kaplan-Meier plots were constructed to show the percentage of animals remaining in the study as a function of time and the same data set was used as a log rank test. All graphics performance and statistical analysis were performed using Prism (GraphPad) of Windows 3.03 (Windows 3.03).

Overview of treatment response

Growth of A375 tumors in control mice (Group 1):

Group 1 animals received 10% cetyl polyoxyethylene ether EL/saline vehicle, po, qd x 14. Tumors in the control group grew to an end point volume of 2000 mm ³ with an intermediate TTE of 22.8 days, and the most likely TC or 163% TGD was established in the 37.1 day study.

The effect of paclitaxel treatment (Group 2):

Group 2 animals were administered with paclitaxel as a reference, 30 mg/kg, iv, qod x 5. 9 animals all reached the tumor volume endpoint. Tumor growth was parallel and slightly shifted to the right compared to the control group. The intermediate TTE value was 28.8 days, corresponding to 26% TGD, with a logarithmic scale analysis as a significant result (Table 2, P = 0.0088 G1 vs. G2). Tumor regression was not associated with paclitaxel treatment.

N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl The effect of cyclopropane-1-sulfonamide treatment (Groups 3-6):

Groups 3-6 accept N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, Oral administration of 3-dihydroxypropyl)cyclopropane-1-sulfonamide as a monotherapy. Group 3 animals were administered 50 mg/kg in qd x 14 time course. Nine tumors in this group reached the end of the volume. The median tumor volume of this group experienced a small net change at the beginning of about 10 days, followed by an increase during the study. A single animal experiences tumor PR. The intermediate TTE value was 27.5 days, or 21% TGD, which was a significant result (P = 0.0054 G1 vs. G).

Group 4 animals received 25 mg/kg of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxy group in a bid x 14 time course Phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. Four of the 9 animals in this group remained on day 60, all of which were TFS. Additional 2/9 animals had tumors reaching the end of the volume one day before the study was terminated. This group has 4/9 PR, 5/9 CR and 4/9 TFS. Median tumor volume decreased for several days from the start of the study and lasted for approximately 30 days. Tumor regrowth in 5/9 animals indicated that the onset of tumor growth was started on day 32 and continued until the study was terminated. The intermediate TTE value for this group was 59.9 days, indicating a maximum probability of 163% TGD (P < 0.0001, Table A1).

Group 5 mice received 25 mg/kg N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( 2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide, but following a lower strength qd x 14 time course. Nine animals in Group 5 reached the tumor volume endpoint and no tumor resolved. Tumor growth in this group and control group was closely followed. The intermediate TTE value was 25.6 days, or 12% TGD, which was a non-significant result ( P = 0.0662 G1 vs. G5).

Group 6 animals were dosed with 12.5 mg/kg N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxy in a bid x 14 time course. Phenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. All tumors in this group reached the end of volume. As in Group 4, the median tumor volume of Group 6 decreased early in the study, but this decrease lasted only about 9 days with a single PR response. Tumor volume increased from day 10 to study termination. The intermediate TTE value for this group was 27.5 days, corresponding to a significant 21% TGD ( P = 0.0424 G1 vs. G6).

Briefly, N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3- Dihydroxypropyl)cyclopropane-1-sulfonamide exhibited dose-related antitumor activity against human A375 melanoma xenografts once daily and twice daily. Secondary dosing daily is superior to once daily dosing in terms of the size of TGD produced and the number of target responses. Thus, N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxyl) The anti-tumor activity of propyl)cyclopropane-1-sulfonamide varies according to the dose and time course.

Example 37: Active animals against subcutaneous COLO205 human colon cancer xenograft:

Female athymic nude mice ( nu / nu , Harlan) were 12 to 13 weeks old and had a body weight (BW) range of 18.3 to 27.3 grams on study day 1. Animal free access to water (reverse osmosis, 1ppm Cl) and NIH31 modification and irradiation Lab It consists of 18.0% crude protein, 5.0% crude fat and 5.0% crude fiber. The mouse is housed in a static micro-separation chamber The laboratory animal pad is in a 12 hour photoperiod with 21-22 ° C (70-72 ° F) and 40-60% humidity. Adhere to the recommendations for restrictions, grazing, surgical procedures, feeding and fluid regulations, and veterinary care for laboratory animal feeding management and use practices.

Tumor transplantation:

Allogeneic transplantation was initiated by COLO205 human colon cancer cells. Tumor cells in 10% heat inactivated fetal bovine serum, 100 units / ml penicillin G sodium, 100 μg / mL streptomycin sulfate, 0.25 μg / mL amphotericin B, 25 μg / mL statamycin, 2 mM branamide Acid, 1 mM sodium pyruvate, 10 mM HEPES and 0.075% sodium bicarbonate were cultured. The cell culture was maintained in a tissue culture flask in a humidified culture tank at 37 ° C, 5% CO ₂ and 95% air atmosphere. In the day of implantation of tumor cells during logarithmic growth and Colo205 cells recovered at 5 × 10 ⁶ cells / mL in PBS and resuspended in a concentration of 50% Matrigel matrix (BD Biosciences). Each test mouse was subcutaneously implanted with 1 x 10 ⁶ Colo205 cells in the right abdomen, and tumor growth was monitored to an average size of 80-120 mm ³ . After 14 days, the day of the study was designated as day 1 and the animals were divided into 8 groups (n=9) with individual tumor volumes ranging from 63 to 196 mm ³ and the average tumor volume of each group was 118-119 mm ³ . Tumor volume was calculated using the following equation: tumor volume (mm ³ ) = (w ² × l) / 2 , where w = COLO205 tumor width (mm) and l = COLO205 tumor length (mm). Tumor weight was estimated by inference of 1 mg equal to 1 mm ³ tumor volume.

material:

The administration solution of Compound A was freshly prepared daily by dissolving the required amount of the compound in 100% cetyl polyoxyethylene ether EL and then diluting it to 10 times with physiological saline. The final dosing solution concentration was 2.5, 5, 10 or 20 mg/mL to provide 25, 50, 100 or 200 mg/kg each dose in a 10 mL/kg dosing volume. Paclitaxel (Natural Pharmaceuticals) was freshly prepared at a daily dose in a vehicle consisting of 5% ethanol and 5% cetyl polyoxyethylene ether EL in 90% D5W (5% EC vehicle).

treatment :

The table below shows the treatment plan.

Group 1 received a formulation vehicle (10% cetyl polyoxyethylene ether EL in saline) and served as a tumor growth control group. Group 2 received the reference drug paclitaxel (30 mg/kg i.v. qod x 5) administered in the optimal time course of nude mice. Groups 3-6 received Compound A at a dose of 25, 50, 100, and 200 mg/kg, respectively, p.o. qd x 14 administered, with Group 6 (200 mg/kg) discontinued after 6 days due to toxicity. All doses were in accordance with animal weight (0.2 mL / 20 g body weight).

end:

Tumors were measured twice a week using calipers. Each animal was euthanized when the tumor reached a predetermined endpoint size of 2000 mm ³ or when either of the last day of the study (day 74) arrived first. However, the control tumor did not exhibit logarithmic growth characteristics after reaching a size of about 800 mm ³ . Therefore, the 800 mm ³ endpoint tumor size was used for the analysis of tumor growth delay (TGD). The endpoint time (TTE) of each mouse is calculated by the following equation: TTE (days) = (log ₁₀ (end point volume, mm ³ )-b)/m, where b is the intercept and m is the log-transformed tumor growth data set The slope of the line obtained by linear regression. This data set contains the first observation that exceeds the study endpoint volume and three consecutive observations immediately before the endpoint volume is reached. Animals that did not reach the end point specified a TTE value equal to the last day of the study. Animals classified as having an accidental NTR (non-therapeutic correlation) (NTRa) or died for unknown reasons (NTRu) were excluded from the TTE calculation (and all further analyses). Animals classified as TR (treatment-related) death or NTRm (non-therapeutic correlation due to metastasis) were assigned a TTE value equal to the day of the study death.

Treatment results were assessed as tumor growth delay (TGD), defined as the increase in endpoint time (TTE) of the treatment group compared to the control group: TGD = TC, expressed in days, or in the middle of the TTE of the control group The percentage indicates: Where: T = intermediate TTE of the treatment group, C = intermediate value of TTE of the control group (Group 1).

The control group was designated as the first group of mice.

Treatment may result in partial regression (PR) or complete regression (CR) of the animal's tumor. PR in the reaction, during the course of the study, tumor volume measurement in three consecutive days of 50% or less of the first volume, and equal to or greater than 13.5mm ³ In this one or more of three measurements . In the CR reaction, tumor volume was less than 13.5 mm ³ in three consecutive measurements during the course of the study. The regression response was monitored and recorded.

side effect

Animals were weighed daily for the first five days of the study and then weighed twice a week. Frequent observations of any adverse, treatment-related side effects of this mouse were observed frequently, and clinical signs of toxicity at the time of observation were recorded. Acceptable tolerance is defined as a group of treatment-related deaths with a mean weight loss of less than 20% during the test and no more than one in the animal group, and any dosing regimen that results in greater toxicity is considered to be greater than the maximum tolerated dose (MTD) ). Death is classified as TR if it is confirmed by medical symptoms and/or anatomy as a side effect of treatment, or TR if it is died due to unknown reasons during administration or during the last 10 days of administration. Death is classified as NTR if there is no evidence that death is associated with treatment side effects. Animals were monitored by frequent observations and BW measurements. BW was changed to be inconspicuous and all treatments were acceptable tolerated, except for Group 6. Compound A once daily p.o. 200 mg/kg dose six times resulted in TR death assessment on day 7 and on day 8 two more TR deaths. All rats in Group 6 exhibited toxic medical symptoms including hunchback, reduced activity, and fecal soft strips.

Statistical and graphical analysis:

Logarithmic scale tests were used to analyze the significance of the difference in TTE values between the treated and control groups. Two-tailed statistical analysis was performed at a significant amount of P = 0.05.

The median value of the tumor growth curve shows that the median tumor volume of each group is a function of time on a logarithmic scale. When the animal withdrew from the study due to tumor size or TR death, the data for the intermediate tumor volume calculated for the subsequent time points included the final tumor volume recorded for the animals. This curve was truncated after 50% of the animals in the group withdrew from the study due to tumor development or after the second TR death in the group. Kaplan-Meier plots were constructed to show the percentage of animals remaining in the study as a function of time and the same data set was used as a log rank test. All graphics performance and statistical analysis were performed using Prism (GraphPad) of Windows 3.03 (Windows 3.03).

Overview of treatment response

Growth of COLO205 tumors in control mice (Group 1)

The first group of tumors showed slow, heterogeneous growth. Tumors of 7/9 vehicle-treated Group 1 control mice reached an end point of 800 mm ³ tumor volume and both mice were maintained until study termination. The median TTE for Group 1 was 41.0 days, and thus the maximum TGD possible for this 74 day study was 33.0 days (80%).

Effect of treatment with paclitaxel (Group 2)

Eight of the paclitaxel-treated mice (n=9) in Group 2 were retained in the study on day 74 with a MTV of 143 ^mm3 . This corresponds to the maximum possible TGD (33.0 days or 80%) and statistically significant activity ( P = 0.002). Five PR responses were recorded. The mean tumor growth curve showed that when tumor growth resumed, MTV decreased on day 19, followed by a slight change until day 47.

Effect of treatment with Compound A (Groups 3-6)

Groups 3, 4, and 5 produced intermediate TTE values of 47.9, 59.1, and 74.0 days, respectively. Groups 3 and 4 had non-significant log-rank results, and the fifth-group log-rank test reached boundary significance ( P = 0.058). These treatments produced a regression in the amount that varied with dose, however, this type of regression response (PR vs. CR) and the number of 74 days of survivors in each group were dose-independent. The median tumor growth curve showed similar activity in the three doses at the early stage of the study (to day 29), followed by a dose-dependent delay in tumor regrowth. Group 6 produced 3 TR deaths and the administration stopped after 6 days. Therefore, treatment at 200 mg/kg was considered to be higher than MTD and could not be used for TGD evaluation.

Compound A illustrates the dose dependent activity of COLO205 colon cancer xenografts. Compound A exhibited a TGD of 3% when administered at 25 mg/kg. Compound A produced 46% TGD at 50 mg/kg. Tolerance to treatment at 100 mg/kg was acceptable and, as with paclitaxel treatment, resulted in maximum TGD possible values in experiments with similar regression numbers. Treatment with 200 mg/kg produced 3/9 TR death and was higher than MTD. Compound A was observed to have a more pronounced initial decrease in tumor burden compared to paclitaxel; however, the duration of this effect was shorter. Tumor regrowth in the 25 and 50 mg/kg groups started at a faster pace than the control group, and MTV approached the control group before the end of the study. Treatment with 100 mg/kg did not show this rapid regrowth, but showed faster tumor growth than paclitaxel.

Example 38: Human Clinical Trial

Randomized, double-blind, open-label, histologically controlled, one-component, safe/effective human Phase I clinical trials were performed with Compound A and placebo at the end of the first chemotherapy or metastatic pancreatic cancer.

The primary objective of this study was to assess the safety and tolerability of Compound A. Secondary outcomes were to assess response rates, clinical benefit, and tumor shrinkage after treatment with Compound A. Furthermore, this study will be designed to assess the timing and overall survival of disease progression in individuals with pancreatic cancer. In addition, DCE-MRI can be used to assess pharmacokinetic changes in tumor vascular parameters including, for example, blood flow, blood volume, peak time ROC-recipient operator characteristics.

Furthermore, biomarkers such as MEK1 and MEK2 genetic polymorphisms and serum proteomics are used to correlate results. This also allows for the determination of the rate of resection of the tumor after treatment and the assessment of the MTD of Compound A.

Compound A will have about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg during the study. , about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, Different doses of about 14 mg, about 14.5 or about 15 mg are administered.

The selection criteria for this study are based on the following factors:

Histology/pathology identified local end-stage unresectable or borderline unresectable pancreatic cancer with no evidence of metastatic disease.

The diagnosis of locally unresectable pancreatic cancer is based on the evaluation of biphasic computed tomography and/or endoscopic ultrasonography (EUS) (described in Annex F, EUS).

The measurable condition according to RECIST was obtained by a two-phase computed tomography scan within 14 days prior to the registration of the treatment proposal.

Tumor size is greater than or equal to 2 cm in a biphasic computed tomography scan.

Appropriate organ function was recorded within 14 days of registration, empirically: absolute neutrophil count >1500/mm ³ ; platelet count; 100,000/mm ³ ; hemoglobin 9 gm/dL, no blood transfusion requirement during the first 4 weeks; total Bilirubin 1.5 times the conventional upper limit (ULN); transaminase (AST and / or ALT) ;PT (or INR) And aPTT is within the normal range (subjects receiving anticoagulant therapy with agents such as warfarin or heparin are allowed to participate); individuals receiving warfarin are monitored at least weekly until the INR is pre-dose Tested as stable, as defined by local care standards; using the Cockcroft-Gault equation to calculate intramuscular guanamine clarity of > 60 ml/min.

Exclusion criteria included: prior treatment with Compound A within 6 months prior to enrollment; clinical confirmation of tumor invasion into the duodenal mucosa (recorded by endoscopic or endoscopic ultrasonography); undergoing minor surgery within 14 days of study enrollment (eg Fine needle extraction or fine needle biopsy); undergo major surgery, significant trauma or severe non-healing wounds, ulcers or fractures within 21 days of study registration; experience any of the following within 6 months prior to study drug administration : severe/unstable angina (symptoms of angina at rest), newly started angina (starting in the last 3 months) or myocardial infarction, congestive heart failure, arrhythmia requiring antiarrhythmia treatment; in the past 6 History of embolism or embolization during the month, such as cerebrovascular accident or transient cerebral ischemia; history of aneurysm or arteriovenous malformation; known human immunodeficiency virus (HIV) infection or chronic B or C hepatitis; active Serious clinical infection is greater than CTCAE level 2; any test drug is accepted within 4 weeks of study registration; despite optimal medical control, hypertension remains uncontrolled It is defined as systolic blood pressure greater than 150 mmHg or diastolic blood pressure greater than 90 mmHg; alveolar hemorrhage/bleeding events greater than CTCAE level 2 within 4 weeks of study registration; any other bleeding/bleeding events greater than CTCAE level 3 within 4 weeks of study registration; The fact or history of bleeding quality or coagulopathy; long-term, daily treatment with aspirin or other non-steroidal anti-inflammatory agents; use of St. John's Wort, rifampin, rifampicin, ketone Accompanying (ketoconazole), itraconazole, ritonavir or grapefruit juice; known or suspected to be allergic to compound A; any condition that impairs the individual's ability to swallow the entire pill; any malabsorption Problem; other serious, acute or chronic medical or mental condition, or may increase the risk of accompanying study participation or study drug administration, or may interfere with the interpretation of the study results, the investigator judges that the individual is not suitable for the study of the study Abnormal examination of the chamber; history of collagen vascular disease; any contraindications to performing nuclear magnetic resonance imaging.

Example 39: Human Clinical Trial

Compound A was randomized, double-blind, open-labeled, histologically controlled, one-component, safe/effective in humans at the end of the first chemotherapy or metastatic gastric cancer in the same manner as specified in Example 117. The first phase of the clinical trial was the exception that the individual involved was diagnosed with gastric lymphoma, gastric stromal tumor or gastric carcinoid tumor.

Example 40: Rat carrageenan-induced paw edema (CPE)

Compound A (6, 20 and 60 mg/kg) or sputum was administered orally 2 hours before the injection of 1% carrageenan suspension into the right hind footpad of male Sprague-Dawley rats (N=6 per treatment group). Indomethacin (3mg/kg). After 3 hours, the paw volume was assessed by plethysmography to measure hind paw edema. A reduction in hind paw edema of 30% or more indicates significant acute anti-inflammatory activity. Indomethacin (Indo) was used as a positive control. Figure 22 shows an increase in paw volume in each treatment group, indicating that oral administration of Compound A resulted in significant anti-inflammatory activity in the rat carrageenan paw edema model in all dose groups.

Example 41: Rat adjuvant arthritis inflammation test

In the rat adjuvant-induced arthritis model, Complete Freund's adjuvant (CFA) was injected into the right hind paw of the rat to induce a lesion similar to rheumatoid arthritis in humans. Compound A was orally administered at 2, 6, and 20 mg/kg for 5 days. Dexamethasone was also orally administered at 5 mg/kg for 5 days. On the first day and the fourth day, 10 mg/kg of enalapril (Enbrel) was administered by subcutaneous injection. CFA was injected into the right hind paw 1 hour after the first dose on day 1. For the acute phase, the percentage of inhibition of the right hind paw swelling on the 1st and 5th days relative to the vehicle-treated control group was determined, and for the lag phase, the left hind paw swelling on the 14th and 18th days was determined as opposed to The percent inhibition of the vehicle treated control group. Multiple arthritis is scored based on the presence of swelling in the front paw, tail, nose or ear.

Figures 23A and 23B show the percent inhibition of swelling in the different treatment groups relative to the control group. Compound A at 20 mg/kg showed a significant reduction in swelling in both the acute and delayed phases. For the multiple arthritis score, 6 animals in the vehicle treatment group had swelling on the forepaw and tail. For the 20 mg/kg Compound A group, 2 out of 6 had no swelling on the forepaws and 4 out of 6 had no swelling on the tail. For the etanercept group, none of the animals were free of swelling on the front paws and 3 of the 6 had no swelling on the tail.

Example 42: Inhibition of collagen-antibody-induced arthritis (CAIA) in mice

Male Balb/c mice (N=8 per treatment group) were injected intravenously (tail vein) with 2 mg of collagen antibody mixture (Chondrex) on day 0. RDEA119 (1, 3 & 10 mg/kg QD) or dexamethasone (1 mg/kg QD) was orally administered from day 0 to day 4, while etanercept was injected subcutaneously on days 1 and 3. All mice were intraperitoneally injected with LPS (50 μg) on day 3 except for the untested animals. Arthritis scores were determined for all limbs and are shown in Figure 24 (maximum score of 16). Significant anti-inflammatory activity was noted for all test articles and reference drugs. The etanerex universal dexamethasone was used as a positive control.

Example 43: In vivo cell proliferation assay

Methods for determining cell proliferation counts of cancer cells treated with MEK protein kinase inhibitors are well known in the art and are described in Kenny, LM et al., Positron Emission Tomography (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. Fifty individuals volunteered to participate in the study, all suffering from pancreatic cancer at a stage similar to cancer development. A combination of Compound A was administered to 25 individuals. Placebo was administered to the last 25 individuals. Each individual is administered a daily dose of radiolabeled tracer (eg, labeled fluoro-2-deoxy-DF-glucose (FDG)) for 14 days.

After 14 days of treatment, trained physicians used non-invasive positron emission tomography (PET) imaging devices to detect tumor cell proliferation. Furthermore, the trained physician will determine cell proliferation counts for both tumors and normal cell tissues of individuals treated with Compound A and placebo. The results indicate a decrease in cell proliferation count between MEK protein kinase inhibitor (eg Compound A) and placebo. This assay for determining cell proliferation counts using labeled tracers and PET imaging is referred to herein as "in vivo cell proliferation methods." Other in vivo methods of cell proliferation are known in the art.

A similar analysis was used to determine the reduction in tumor size.

Example 44: In vivo cell apoptosis assay

MEK inhibitors (e.g., Compound A) are tested in vivo to determine their effect on apoptosis of cancer cells. Forty individuals volunteered to participate in the study, all suffering from pancreatic cancer at a stage similar to cancer development. Compound A was administered to 20 individuals and placebo was administered to 20 individuals. The daily dose was administered to each individual for 14 days.

After 14 days, each individual will consume a detectable lipopolysaccharide binding protein (LBP) reagent coupled to a label. According to WO/2006/054068 (which is incorporated herein by reference in its entirety), each individual is in the scanning range of the scanning device, whereby the scanning device detects a consumable agent that binds to dead cells. The number of dead cells is associated with the degree of cellular apoptosis in each individual. The degree of apoptosis in the individual administered with the combination is compared to the degree of apoptosis in the individual administered the single entity agent and compared to the group administered the placebo. The method for detecting the degree of apoptosis by using a lipopolysaccharide-binding protein and a scanning device is referred to herein as "in vivo apoptosis".

Example 45: Dissolution study

Capsules containing Compound A were prepared as in the above examples. The following dissolution data was obtained using the USP <711> dissolution method.

Figure 1 shows a 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 cell. The mice were orally administered once a day (25 mg/kg, 50 mg/kg or 100 mg/kg) for 14 days;

Figure 2 shows a graph of % tumor growth inhibition (% TGI) in A375 xenografted mice administered 50 mg/kg QD, 25 mg/kg BID, 50 mg/kg QD, and 12.5 mg/kg BID dose;

Figure 3 is a graph showing the relationship between plasma concentration (log nM) and % inhibition of pERK in female nu/nu mice transplanted with Colo205 tumor cells. The mice are given a single dose of 2.5, 5, 10 or 25 mg/kg;

Figure 4 is a graph showing the relationship between plasma concentration (ng/mL) and time (hour) after administration of a single dose of 2 mg (2 x 1 mg capsule), 4 mg (4 x 1 mg capsule) or 6 mg (6 x 1 mg capsule) in humans;

Figure 5 is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di A pattern of powder x-ray diffraction (PXRD) spectra of hydroxypropyl)cyclopropane-1-sulfonamide Form A, which was produced using an Inel XRG-3000 diffractometer. This plot is plotted with respect to the diffraction angle 2θ (°) as defined by the count per second;

Figure 6 is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxybenzene produced using TA Instruments Differential Scanning Calorimeter Q1000 Differential Scanning Calorimetry (DSC) Modulation Thermogram of Form I-1-(2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide Form A. This graph plots the measured sample temperature (°C) with the corrected heat flux (in watts/gram, W/g);

Figure 7 is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-di Hydroxypropyl)cyclopropane-1-sulfonamide Form A (upper) and N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6- A plot of the PXRD pattern of the amorphous (bottom) methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, which was produced using an Inel XRG-3000 diffractometer. This graph plots the diffraction intensity defined by the count per second versus the diffraction angle 2θ (°);

Figure 8 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxybenzene produced using a VTI SGA-100 vapor adsorption analyzer. Dynamic vapor sorption/desorption (DVS) isotherm of Form A-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide Form A;

Figure 9 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl which was produced using a TA Instrument 2950 thermogravimetric analyzer. Thermogram analysis (TG) thermogram of Form-1 of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide;

Figure 10 (a) and Figure 10 (b) show exposure to increasing concentrations of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6- The growth of the logarithmic phase-cleaved A375 cells of oxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was arrested. Analyze the ATP content of the cells. Using 1 μM N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxyl Propyl)cyclopropane-1-sulfonamide assay for 100% growth arrest;

Figure 11 shows the 48 hour AK assay in A375 cells. Logarithmic phase-cleaved A375 cells were exposed to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( 2,3-Dihydroxypropyl)cyclopropane-1-sulfonamide and PD-325901 for 48 hours and analyzed for AK release;

Figures 12A-12C show N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3 - dihydroxypropyl) cyclopropane-1-sulfonamide Amides of (A) human growth of colorectal cancer Colo205 cells (GI ₅₀ = 11nM) of inhibition; growth (B) A375 cells (GI ₅₀ = 22nM) and inhibition of ( C) Inhibition of MDA-MB231 cells, which did not show N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino) in a two-dimensional fixation-dependent assay - 6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide - causing growth arrest;

13A shows the inhibition of growth of human colorectal cancer in the Colo205 cells, and ₅₀ and the values in the lower GI 11nM 6nM respectively;

Figure 13B shows inhibition of growth of A375 cells as well as in the GI ₅₀ values of 5nM and 22nM lower;

14A and 14B show N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2, Effect of 3-dihydroxypropyl)cyclopropane-1-sulfonamide on cell cycle progression, showing exposure of A375 cells to N-(S)-(3,4-difluoro-2-(2-fluoro-4) -iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide causes stagnation of the G1 phase of the cell cycle, which is caused by G2 And the indication of cell consumption during the S phase;

15A and 15B show exposure to N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-( N-(S)-(3,4-difluoro-2-(2-) after 2 days (Fig. 15A) and 6 days (Fig. 15B) of 2,3-dihydroxypropyl)cyclopropane-1-sulfonamide Fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide for gastric cancer (stomach adenocarcinoma) cell line AGS influences. The y-axis is the number of cells relative to the vehicle and the x-axis is N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxybenzene μM of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide;

Figure 16 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) in tumor-bearing mice. -1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide (2 mg/kg once a day, orally (po); once daily 10 mg/kg, orally and once daily 50 mg/kg, orally) Average liver weight after;

Figure 17 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl) in tumor-bearing mice. 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide (2 mg/kg once a day, orally; 10 mg/kg once daily, 50 mg/kg once daily, orally) Tumor weight

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

Figure 19 shows the number of cells (relative to vehicle) versus N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl Inhibition of proliferation of Hs746t cells in a graph of the concentration of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide;

Figure 20A depicts the comparison of N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1 at increasing concentrations. -(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was used to treat the degree of apoptosis of individual cells on day 5 of non-small cell lung cancer (NSCLC) MV522 cells;

Figure 20B depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of individual cells on day 5 of non-small cell lung cancer (NSCLC) H358 cells treated with (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide;

Figure 20C depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on the 6th day of treatment of non-small cell lung cancer (NSCLC) A549 cells;

Figure 20D depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on the 5th day of treatment of non-small cell lung cancer (NSCLC) H727 cells;

Figure 20E depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on the 5th day of treatment of colonic HT29 cells;

Figure 20F depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of the respective cells on day 6 of treatment of colonic HCT116 cells with (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide;

Figure 20G depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of the respective cells on day 5 of treatment of colonic HUH7 hepatoma cells with (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide;

Figure 20H depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on the 5th day of treatment of sarcoma U2-OS cells;

Figure 20I depicts N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1- at increasing concentrations The degree of apoptosis of (2,3-dihydroxypropyl)cyclopropane-1-sulfonamide on the 5th day of treatment of glioma D37 cells;

Figure 21 shows the selectivity of Compound A for MEK1 and MEK2 relative to a panel of 205 enzymes (10 μM). The cell lines are Colo205, A375, A431 and HT-29;

Figure 22 is a graph showing the administration of N-(S)-(3,4-difluoro-2-(2-fluoro-) to rats at 6, 20, 60 and 200 mg/kg in a rat model of carrageenan paw edema. After 4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, the paw volume increased in each treatment group and a graph with reduced edema relative to the vehicle control group;

Figure 23A shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodobenzene) at 2, 6 and 20 mg/kg in the acute phase in an adjuvant-induced arthritis model. Swelling of rats treated with arylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was inhibited;

Figure 23B shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodobenzene) at 2, 6, and 20 mg/kg in the adjuvant-induced arthritis model. Swelling of rats treated with arylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide was inhibited;

Figure 24 shows N-(S)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl group at 1, 3 and 10 mg/kg QD. Mean arthritis score of collagen-antibody-induced arthritis (CAIA) mice treated with 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide.

(no component symbol description)

Claims (27)

N-(-)-(3,4-Difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropane) Crystalline polymorph A of cyclopropane-1-sulfonamide having substantially the same powder x-ray diffraction pattern as the powder x-ray diffraction pattern shown in Figure 5, or as shown in the figure The differential scanning calorimetry spectrum of the differential scanning calorimetry spectrum in 6 is substantially the same. A crystalline polymorph of claim 1 having an initial melting point of about 143 ° C as measured by differential scanning calorimetry. The crystalline polymorph of claim 1 which is substantially anhydrous. A crystalline polymorph of claim 2 which is substantially anhydrous. A crystalline polymorph of claim 1 or 3 which is substantially solvent free. A crystalline polymorph of claim 2 which is substantially solvent free. A crystalline polymorph of claim 4 which is substantially solvent free. The crystalline polymorph of claim 1 which comprises crystalline amorphous N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxybenzene Prepared by the method of the step of 1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide. A pharmaceutical composition comprising an effective amount of the crystalline polymorph of any one of claims 1 to 8 and at least one excipient or carrier. A use of the polymorph of any one of claims 1 to 8 for the preparation of a medicament, wherein the medicament is for treating the disorder in an individual suffering from a MEK-mediated disorder. The use of claim 10, wherein the MEK-mediated condition is selected from the group consisting of an inflammatory condition, an infectious condition, an autoimmune condition, a stroke, an ischemia, a cardiac condition, a neurological condition, a fibrotic condition, a proliferative condition, A group of hyperproliferative disorders, tumors, leukemias, neoplasms, cancers, cancers, metabolic diseases, and malignant diseases. The use of claim 11, wherein the MEK-mediated condition is a proliferative disorder. The use of claim 12, wherein the proliferative disorder is cancer, psoriasis, restenosis, autoimmune disease, or atherosclerosis. The use of claim 13, wherein the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma, prostate cancer, renal cancer, colorectal cancer, leukemia, myeloid leukemia, glioblastoma, Follicular lymphoma, negative pre-B cells (Pre-B) acute leukemia, chronic lymphocyte B-leukemia, gastric cancer, mesothelioma or small cell lung cancer. The use of any one of claims 11 to 14, wherein the agent is for use in combination with at least one therapeutic agent or at least one additional cancer therapy selected from the group consisting of radiation therapy, non-MEK kinase inhibitor therapy, chemotherapy, Surgical procedures, glucocorticoids, methotrexate, bioreactive modifiers, or combinations thereof. The use of claim 15, wherein the therapeutic agent is sorafenib or gemcitabine. The use of claim 16, wherein the agent is for use with sorafenib for the treatment of hepatocellular carcinoma. The use of claim 16, wherein the agent is for use with gemcitabine for the treatment of pancreatic cancer. Use of the composition of claim 9 for the preparation of a medicament, wherein The agent is for the treatment of the condition in an individual suffering from a MEK mediated condition. The method of claim 19, wherein the MEK-mediated condition is selected from the group consisting of an inflammatory condition, an infectious condition, an autoimmune condition, a stroke, an ischemia, a cardiac condition, a neurological condition, a fibrotic condition, a proliferative disorder, A group of hyperproliferative disorders, tumors, leukemias, neoplasms, cancers, cancers, metabolic diseases, and malignant diseases. The use of claim 20, wherein the MEK-mediated condition is a proliferative disorder. The use of claim 21, wherein the proliferative disorder is cancer, psoriasis, restenosis, autoimmune disease, or atherosclerosis. The use of claim 22, wherein the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma, prostate cancer, renal cancer, colorectal cancer, leukemia, myeloid leukemia, glioblastoma, Follicular lymphoma, negative pre-B cells (Pre-B) acute leukemia, chronic lymphocyte B-leukemia, gastric cancer, mesothelioma or small cell lung cancer. The use of any one of claims 19 to 23, wherein the agent is for use in combination with at least one therapeutic agent or at least one additional cancer therapy selected from the group consisting of radiation therapy, non-MEK kinase inhibitor therapy, chemotherapy, Surgical procedures, glucocorticoids, methotrexate, bioreactive modifiers, or combinations thereof. The use of claim 24, wherein the therapeutic agent is sorafenib or gemcitabine. The use of claim 25, wherein the agent is for use with sorafenib for the treatment of hepatocellular carcinoma. The use of claim 25, wherein the agent is for use with gemcitabine for the treatment of pancreatic cancer.