EA020624B1 - Crystalline polymorph form a of n-(-)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-l-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide and use thereof as inhibitor of mek - Google Patents

Abstract

The present invention relates to a crystalline polymorph form A of N-(-)-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide, which exhibit a specific powder x-ray diffraction profile and/or a specific differential scanning calorimetry profile. The present invention also relates to pharmaceutical compositions comprising the compounds described herein and methods of use of the compounds and compositions described herein, including the use in the treatment and/or prevention of cancer, hyperproliferative diseases and inflammatory conditions. The invention also relates to methods of making the compounds and compositions described herein.

Description

The present invention relates to a crystalline polymorphic form ABY - (-) - (3,4-difluoro-2 (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1- sulfonamide, which is an MEK inhibitor, exhibiting a specific profile of a powder x-ray and / or a specific profile of differential scanning calorimetry. The present invention also relates to pharmaceutical compositions that contain a compound described in this invention, and methods of using the compounds and compositions described in this invention, including use for the treatment and / or prevention of malignant neoplasms, hyperproliferative diseases and inflammatory conditions. The invention also relates to methods for producing the compounds and compositions described in this invention.

BACKGROUND OF THE INVENTION

Oncogenes - genes that are involved in the formation of malignant neoplasms are usually mutated forms of normal normal cell genes (proto-oncogenes). Oncogenes often encode abnormal versions of signaling pathway components such as receptor tyrosine kinases, serine-threonine kinases, or downstream signaling molecules. The central downstream signaling molecules are Like proteins, which are anchored on the inner surface of the cytoplasmic membranes, and which hydrolyze bound guanosine triphosphate (OTP) to guanosine triphosphate (SER). When activated by a growth factor, growth factor receptors initiate a chain of reactions that lead to activation of the exchange activity of guanine nucleotides on How. How it alternately switches between the active state turned on with the associated OTP (hereinafter, referred to as OTP) in the present invention and the inactive state turned off with the associated SIR. An active on state, Like OTR, binds to and activates proteins that control cell growth and differentiation.

For example, in a cascade of mitogen-activated protein kinase (tyodep-as11ua1 prog! Esh ktake (MAP kshake)) As OTR leads to activation of the cascade of serine / threonine kinases. One of the several groups of kinases for which the need for How to. OTP for their own activation is known is Ka! family. Ka! proteins activate MEK1 and MEK2, abbreviations for mitogen-activated EKK-activating kinases (tyodep-as11ua1eb EKK-asIuaIpd kzpakek) (where EKK is a protein kinase regulated by extracellular signals, another designation for MAPK). MEK1 and MEK2 are dual-function serine / threonine and tyrosine protein kinases and are also known as MAP kinases. Thus, OTP activates Ka !, which activates MEK1 and MEK2, which activate the MAP kinase (MAPK). It is believed that the activation of MAP kinase by mitogens is important for proliferation, and the constitutive activation of this kinase is sufficient to induce cell transformation. Blocking the downstream As signal transduction, as carried out using a dominant negative Ka! -1 protein, can completely inhibit mitogenesis, either due to induction from cell surface receptors or from oncogenic As mutants.

Interaction Ka! and As a key regulatory step in controlling cell proliferation. To date, no other MEK substrates have been identified except for MAPK; however, recent studies have shown that MEK can also be activated by other upstream signaling proteins, such as MEK kinase or MEKK1 and PKC. Activated MAPK moves and accumulates in the nucleus, where it can phosphorylate and activate transcription factors, such as E1k-1 and 8ap1a, which leads to enhanced gene expression, such as, for example, for c-! Ok.

When activated Ka! and other kinases phosphorylate MEK at two adjacent serine residues,

218 222 and 8 in the case of MEK1. These phosphorylation are necessary for the activation of MEK as a kinase. In turn, MEK phosphorylates MAP kinase at two residues separated by a single amino acid: tyrosine, Υ ¹⁸⁵ , and threonine, T ¹⁸³ . It is believed that MEK is tightly bound to MAP kinase before phosphorylation, suggesting that prior strong interaction between the two proteins may be necessary for phosphorylation of MAP kinase using MEK. Two factors - the unusual specificity of MEK and its need for strong interaction with MAP

- 1,020,624 kinase before phosphorylation - confirm the fact that the MEK-mechanism of action can differ significantly from the mechanisms of other protein kinases, which makes it possible to detect selective MEK inhibitors. Perhaps such inhibitors will act through allosteric mechanisms, and not through a more conventional mechanism that involves blocking the ATP binding site.

Thus, MEK1 and MEK2 are reasonable and recognized targets for antiproliferative therapies, even if the oncogenic mutation does not affect the structure or expression of MEK. See, for example, published patent applications I82003 / 0149015 (inventors Waggay and others) and I82004 / 0029898 (inventors Wowl and others).

Several examples of 1-substituted-2- (p-substituted-phenylamino) aryl MEK inhibitors are described. Patents и 86440966 and 8 86750217 and the corresponding publication UO 00/42003 describe esters of carboxylic and hydroxamic acids and Ν-substituted amide derivatives of sulfonamide-substituted 2- (4-iodophenylamino) benzoic acid esters and Ν-substituted benzamides, which act in as MEK inhibitors. Sulfonamide may also be Ν-substituted.

Patent I86545030 and the corresponding publication UO 00/42029 describe MEK inhibitors, which are 1-heterocyclyl-2- (4-iodophenylamino) benzene, where the heterocycle is a five-membered nitrogen-containing ring, such as pyrazole, triazole, oxazole, isoxazole and isoxazolinone . A later published patent application I82005 / 004186 describes related compounds in which the 4-iodine substitute of I86545030 is replaced by a very large class of components such as alkyl, alkoxy, acyloxy, alkenyl, carbamoyl, carbamoylalkyl, carboxyl, carboxylalkyl, Νacylsulfonamide.

Patent I86469004 and the corresponding publication \ UO 00/42022 describe esters of carboxylic and hydroxamic acids of the group of heterocyclo-condensed phenylene compounds, i.e. benzimidazoles, benzooxazoles, benzothiazoles, benzothiadiazoles, quinazolines, etc. Heterocycles are 7-P-6- (4-iodophenylamino) -5-carboxylic acid esters, carboxylic acid amides or hydroxamic acid esters. Later publications I8 2005/0026970 describe similar compounds in which the 4-iodine substituent is replaced by a very large class of structures. Related compounds are described in published patent applications \ UO 03/077855, \ UO 03/77914 and I82005 / 0554701. Further examples of 2- (4-iodophenylamino) phenylhydroxamic acid esters, which are described as suitable as MEK inhibitors, can be found in UO 2005/028426.

The published patent application UO 02/06213 and the corresponding application I810 / 333399 (L82004 / 0054172) describe hydroxy substituted acid esters of 1-oxamic acid - 2- (4halophenylamino) -3,4-difluorobenzene. Patent I86891066 and the corresponding publication UO 03/62191 describe similar compounds in which the 4-halosubstituent is replaced by a very large class of structures. Substituents in the 4th position are methyl, ethyl, ethynyl and 2-hydroxyethyl. Specific similar compounds are described in I86770778.

In patent application \ UO 04/083167 (in Japanese), published on September 30, 2004, more than two thousand are described - but NMR data are presented for only 400 - 1- (L-substituted sulfonyl urea) -2- (2,4 -digalophenylamino) -3,4-difluorobenzenes and it is believed that they are suitable as MEK inhibitors. Data indicating inhibition of MEK are presented for only a subgroup of twelve compounds. In addition to the secondary or tertiary amine, these twelve compounds all contain one of the following groups: Ν, Ν-disubstituted sulfonyl urea, Ν-piperazinesulfonamide, Ν-piperidinesulfonamide or Ν-pyrrolidinesulfonamide.

The MEK cascade is also involved in inflammatory diseases and disorders; in the published patent application I82006 / 0030610 (inventors Koy and others), published patent application I82006 / 0140872 (inventors Ritie and others). They include both acute and chronic inflammatory disorders. Examples of such disorders are allergic contact dermatitis, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, chronic obstructive pulmonary disease, psoriasis, multiple sclerosis, asthma, diseases and disorders associated with complications of diabetes, and inflammatory complications of the cardiovascular system, such as acute coronary syndrome. Inflammatory bowel diseases are Crohn's disease and ulcerative colitis.

MEK1 and MEK2 are reasonable and recognized targets for antiproliferative therapies, even if the oncogenic mutation does not affect the structure or expression of MEK. See, for example, published patent applications I82003 / 0149015 (inventors Waggay and others) and I82004 / 0029898 (inventors Wowl and others).

SUMMARY OF THE INVENTION

The present invention relates to the crystalline polymorphic form A ^ (-) - (3,4-difluoro-2 (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide which has a X-ray powder diffraction pattern comprising at least 50% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5.

Preferably, this crystalline polymorphic Form A has a X-ray powder diffraction pattern that contains at least 70%, in particular at least 90% of the peaks identified on the 2 X-ray powder diffraction pattern shown in FIG. 5.

More preferably, this crystalline polymorphic form A has an X-ray powder diffraction pattern substantially identical to the X-ray powder diffraction pattern shown in FIG. 5.

Compound I - (-) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1-sulfonamide is also referred to in the present invention as Compound A or L- (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

Compound A is characterized as the B isomer by producing K- and B-ΜΤΡΑ esters on a secondary alcohol and comparing the differences in the chemical shift of the proton. See, for example, Ea1c. TA .; Mokyeg, N.B., 1. At. Siet. Fight., 1973, 95, 512 and Oy! At et al., 1. At. Siet. Fight., 1991, 113, 4092.

The invention also relates to a crystalline polymorphic form A Y- (B) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

which exhibits a specific differential scanning calorimetry pattern, which is substantially identical to the differential scanning calorimetry pattern shown in FIG. 6.

Preferably, the crystalline polymorph has a melting point that appears, as determined by differential scanning calorimetry, at about 143 ° C.

Preferably, the crystalline polymorph is substantially free of water.

Preferably, the crystalline polymorph is substantially free of solvent.

The invention also relates to pharmaceutical compositions that contain an effective amount of crystalline polymorphic form A Y- (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3- dihydroxypropyl) cyclopropane-1-sulfonamide; and a pharmaceutically acceptable carrier or excipient.

Preferably, the crystalline polymorphic form A of L- (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide is suitable for the treatment of or preventing a malignant neoplasm or inflammatory disease.

Preferably, the pharmaceutical compositions further comprise a pharmaceutically acceptable carrier. Such compositions may contain adjuvants, excipients and preservatives, substances for slowing absorption, fillers, binders, adsorbents, buffers, disintegrating substances, solubilizing components, other carriers and other inert components. Methods for preparing such compositions are well known in the art.

Preferably, the pharmaceutical composition is in a form suitable for oral administration. Preferably, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release preparation, solution, suspension, for parenteral injection in the form of a sterile solution, suspension or emulsion, for topical administration as an ointment or cream, or for rectal administration as a suppository . Preferably, the pharmaceutical composition is in unit dosage forms suitable for single administration of exact doses. Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.002 to about 6 g / day. Preferably, the amount of active compound is from about 0.005 to about 5 g / day. Preferably, the amount of active compound is from about 0.01 to about 5 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably the amount of active compound

- 3,020,624 is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day.

Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary. Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. Preferably, the active compound is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day.

Preferably, the pharmaceutical composition is for administration to a mammal. Preferably, the mammal is a human.

Preferably, the pharmaceutical composition further comprises a pharmaceutical carrier, excipient and / or adjuvant. Also, the pharmaceutical composition may further comprise at least one therapeutic agent which is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents and anti-tumor agents. For example, an antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is taxol, botresomib, or both. Also, the pharmaceutical composition can be used in combination with adjunctive therapy, which is radiation therapy, chemotherapy, surgery, or any combination thereof.

The proposed polymorphic form can be used to treat malignant neoplasms or inflammation using compositions containing approximately 1-100 mg of the active compound. Preferably, the composition modifies the release of the compound. For example, the composition can slow the release of the compound. Preferably, the composition allows delaying the release of the compound. Preferably, the compound is present in an amount of about 1-50 mg. More preferably, the compound is present in an amount of about 1-10 mg. Preferably, the compound is present in an amount of about 10-20 mg. Preferably, the compound is present in an amount of about 20-40 mg. Preferably, the compound is present in an amount of about 40-50 mg.

Also, the proposed polymorphic form can be used to treat malignant neoplasms or inflammation using compositions containing approximately 1-50 mg of the active compound, where the composition allows the release of the drug to be modified. Preferably, the composition further comprises microcrystalline cellulose. Preferably, the composition further comprises croscarmellose sodium. Preferably, the composition further comprises sodium lauryl sulfate. Preferably, the composition further comprises magnesium stearate.

The composition may contain approximately 1 mg of the active compound having the following structure:

Preferably, such a composition further comprises approximately 222.2 mg of microcrystalline cellulose. Preferably, the composition further comprises approximately 12.0 mg of croscarmellose sodium. Preferably, the composition further comprises about 2.4 mg of sodium lauryl sulfate. Preferably, the composition further comprises about 2.4 mg of magnesium stearate.

Also, the proposed polymorphic form can be used for the treatment of malignant neoplasms or inflammation using compositions containing approximately 10 mg of a compound having the following structure:

- 4,020,624

Preferably, such a composition further comprises about 213.2 mg of microcrystalline cellulose. Preferably, the composition further comprises approximately 12.0 mg of croscarmellose sodium. Preferably, the composition further comprises about 2.4 mg of sodium lauryl sulfate. Preferably, the composition further comprises about 2.4 mg of magnesium stearate.

Also, the proposed polymorphic form can be used for the treatment of malignant neoplasms or inflammation using compositions containing approximately 20 mg of a compound having the following structure:

Preferably, such a composition further comprises about 203.2 mg of microcrystalline cellulose. Preferably, the composition further comprises approximately 12.0 mg of croscarmellose sodium. Preferably, the composition further comprises about 2.4 mg of sodium lauryl sulfate. Preferably, the composition further comprises about 2.4 mg of magnesium stearate.

Also, the proposed polymorphic form can be used for the treatment of malignant neoplasms or inflammation using compositions containing approximately 40 mg of a compound having the following structure:

Preferably, such a composition further comprises about 183.2 mg of microcrystalline cellulose. Preferably, the composition further comprises approximately 12.0 mg of croscarmellose sodium. Preferably, the composition further comprises about 2.4 mg of sodium lauryl sulfate. Preferably, the composition further comprises about 2.4 mg of magnesium stearate.

Also, the proposed polymorphic form can be used for the treatment of malignant neoplasms or inflammation using compositions containing approximately 0.4 wt.% Of a compound having the following structure:

5,020,624 and approximately 99.6% by weight of a pharmaceutically acceptable carrier or excipient. Preferably, the pharmaceutically acceptable carrier or excipient contains microcrystalline cellulose. Preferably, microcrystalline cellulose comprises about 92.6% by weight of the composition. The composition may further comprise about 5 wt.% Croscarmellose sodium. Also, the composition may further comprise about 1 wt.% Sodium lauryl sulfate. In addition, the composition may further comprise about 1 wt.% Magnesium stearate.

Also, the proposed polymorphic form can be used for the treatment of malignant neoplasms or inflammation using compositions containing approximately 4.2 wt.% Of the compound having the following structure:

and approximately 95.8% by weight of a pharmaceutically acceptable carrier or excipient. Preferably, the pharmaceutically acceptable carrier or excipient contains microcrystalline cellulose. Preferably, microcrystalline cellulose comprises about 88.8% by weight of the composition. The composition may further comprise about 5 wt.% Croscarmellose sodium. Also, the composition may further comprise about 1 wt.% Sodium lauryl sulfate. In addition, the composition may further comprise about 1 wt.% Magnesium stearate.

Also, the proposed polymorphic form can be used for the treatment of malignant neoplasms or inflammation using compositions containing from about 2 to about 10 wt.% A compound having the following structure:

and from about 98 to about 90 wt.% a pharmaceutically acceptable carrier or excipient. Preferably, the pharmaceutically acceptable carrier or excipient further comprises microcrystalline cellulose. Preferably, microcrystalline cellulose comprises from about 85 to about 95% by weight of the composition. The composition may further comprise from about 1 to about 6 wt.% Croscarmellose sodium. Also, the composition may further comprise from about 0.1 to about 2 wt.% Sodium lauryl sulfate. In addition, the composition may further comprise from about 0.25 to about 1.5 wt.% Magnesium stearate.

The polymorphic form A - - (-) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide can be obtained using a process comprising the crystallization step of amorphous L- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide. Preferably, the crystallization step comprises crystallization from a mixture of ethyl acetate and heptane. Preferably, the mixture of ethyl acetate and heptane contains in a quantitative ratio from about 1-4 parts of ethyl acetate to about 2-10 parts of heptane. Preferably, the mixture of ethyl acetate and heptane contains in a quantitative ratio from about 2 parts of ethyl acetate to about 5 parts of heptane.

The proposed polymorphic form can be used to inhibit MEK enzymes by contacting said MEK enzyme with a compound or composition described in this invention, where the compound is present in an amount sufficient to inhibit said enzyme by at least 25%. Preferably, the MEK enzyme is a MEK kinase. Preferably, said contacting is carried out in a cell.

Also, the proposed polymorphic form can be used to treat an MEK-mediated disorder in an individual suffering from said disorder by administering to said individual an effective amount of a compound or composition as described herein. Preferably, the MEK inhibitor is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, treatment with an inhibitor of a different kinase other than MEK, chemotherapy, surgery, glucocorticoid, methotrexate, biological response modifiers, or any combination thereof. Preferably, the MEK-mediated disorder is selected from the group consisting of inflammatory diseases, infections, autoimmune disorders, stroke, ischemia, cardiac disturbance, neurological disorders, fibrogenetic disorders, proliferative disorders, hyperproliferative disorders, tumors, leukemia, neoplasms, malignant neoplasms, carcinomas, metabolic diseases and malignant diseases. Preferably, the MEK-mediated disorder is a hyperproliferative disease. Preferably, the MEK-mediated disorder is a malignant neoplasm, tumor, leukemia, neoplasm or carcinoma. Preferably, the MEK-mediated disorder is an inflammatory disease. Preferably, said inflammatory disease is rheumatoid arthritis or multiple sclerosis.

Also, the proposed polymorphic form can be used to treat or prevent a proliferative disease in an individual by administering to said individual an effective amount of a compound or composition as described in this invention. Preferably, the proliferative disease is a malignant neoplasm, psoriasis, restenosis, disease, or atherosclerosis. Preferably, the proliferative disease is a malignant neoplasm. In some embodiments, the malignant neoplasm is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, colon and rectal cancer, leukemia, myelogenous leukemia, glioblastoma, follicular lymphoma, acute leukemia B cell precursors, chronic B-cell lymphocytic leukemia, gastric cancer, mesothelioma, or small cell lung cancer. Preferably, at least one further therapeutic agent may be added. Preferably, at least one additional cancer therapy, which is radiation therapy, treatment with an inhibitor of a different kinase other than MEK, chemotherapy, surgery, glucocorticoid, methotrexate, biological response modifiers, or any combination thereof, can be used.

Also, the proposed polymorphic form can be used to treat or prevent an inflammatory disease in an individual by administering to said individual an effective amount of a composition that contains a compound as described in this invention. Preferably, the inflammatory disease is rheumatoid arthritis or multiple sclerosis.

Also, the proposed polymorphic form can be used to destroy, inhibit the growth or destruction of cancer cells by contacting the cells with as many compounds or compositions as described in this invention, which is effective for destruction, inhibiting the growth or destruction of cancer cells. Preferably, the cancer cells include cancer cells of the brain, breast, lung, ovary, pancreas, prostate, kidney, stomach or colon and rectum.

Also, the proposed polymorphic form can be used to inhibit an increase in tumor size, reduce tumor size, reduce tumor proliferation, or prevent tumor proliferation in an individual by administering to said individual an effective amount of a compound or composition as described herein to inhibit tumor increase in size, reduce size tumors, reducing tumor proliferation or preventing tumor proliferation. Preferably, the tumor occurs in the brain, mammary gland, lungs, ovaries, pancreas, prostate, kidneys, stomach, colon or rectum.

Also, the proposed polymorphic form can be used to treat or prevent ankylosing spondylitis, gout, tendonitis, bursitis or sciatica.

The proposed polymorphic form can be used to treat gastric cancer by administering a therapeutically effective amount of a compound or composition as described in this invention. Also, the proposed polymorphic form can be used to treat leukemic melanoma or hepatoma by administering a therapeutically effective amount of a compound or composition, as described in this invention.

The proposed polymorphic form can be used to treat non-small cell lung cancer by administering a therapeutically effective amount of a compound or composition as described in this invention. The proposed polymorphic form can also be used to treat colon cancer by administering a therapeutically effective amount of a compound or composition, as described in this invention. The proposed polymorphic form can also be used to treat CNS cancer by administering a therapeutically effective amount of a compound or composition as described in this invention. The proposed polymorphic form can also be used to treat ovarian cancer by administering a therapeutically effective amount of a compound or composition as described in this invention. The proposed polymorphic form can also be used to treat kidney cancer by administering a therapeutically effective amount of a compound or composition, as described in this invention. The proposed polymorphic form can also be used to treat prostate cancer by administering a therapeutically effective amount of a compound or composition, as described in this invention. The proposed polymorphic form can also be used to treat breast cancer by administering a therapeutically effective amount of a compound or composition, as described in this invention. Preferably, at least one further therapeutic agent may be added. It is also preferred that at least one additional anti-cancer therapy is performed, which is radiation therapy, chemotherapy, surgery, or any combination thereof.

Also, the proposed polymorphic form can be used to treat or prevent psoriasis by administering a therapeutically effective amount of a compound or composition as described herein in a local dosage form.

Preferably, the compositions are administered orally. Preferably, the composition is administered once a day or twice a day. Preferably, the composition is administered once a day for at least one week.

Preferably, when the composition is orally administered, the T _{max of the} compound is achieved between 1 and 3 hours after the composition is administered to the suffering subject. Preferably, when administered to a subject, the compound reaches C _max in the range of from about 0.01 to about 1.0 μg / ml per day 1. Preferably, when administered to the subject, the compound reaches C _max in the range of from about 0.01 to about 0.8 μg / ml per day 1. Preferably, when administered to a subject, the compound reaches C _max in the range of about 0.03 to about 0.5 μg / ml per day 1. Preferably, the compound has LIS in the range of about 0.1 to about 5.0 μg-h / ml for 0-12 hours. Preferably soy Inonii has LIS in the range from about 0.1 to about 4.0 micrograms-hour / mL. Preferably, the compound has an LIS in the range of about 0.5 to about 3.0 μg-h / ml. Preferably, the compound has a T _max in the range of 0.5 to 5.0 hours. Preferably, the compound has a T _max in the range of 1.0 to 3.0 hours. Preferably, the compound has a T _max in the range of 1.0 to 2.5 hours. Preferably, the compound has a plasma concentration of greater than about 0.01 mg / ml 5 hours after a single dose. Preferably, the compound has a plasma concentration of greater than about 0.01 mg / ml 10 hours after a single dose. Preferably, the compound has a plasma concentration of greater than about 0.01 mg / ml 15 hours after a single dose.

Preferably, when administered to a group of 10 subjects, the compound reaches an average C _max in the range of from about 0.01 to about 1.0 μg / ml per day 1. Preferably, when administered to a group of 10 subjects, the compound reaches an average C _max in the range of from about 0.01 up to about 0.8 μg / ml per day 1. Preferably, when administered to a group of 10 subjects, the compound reaches an average C _max in the range of about 0.03 to about 0.5 μg / ml per day 1. Preferably, the compound has an average LIS of interval from from about 0.1 to about 5.0 μg-h / ml Preferably, the compound has an average LIS value in the range of about 0.1 to about 4.0 μg-h / ml. Preferably, the compound has an average LIS in the range of from about 0.5 to about 3.0 μg-h / ml. Preferably, the compound has an average T _max in the range of 0.5 to 5.0 hours. Preferably, the compound has an average T _max in the range of 1.0 to 3.0 hours. Preferably, the compound has an average T _max in the range of 1.0 to 2 5 hours

Also, the proposed polymorphic form can be used to reduce the volume of the tumor by introducing compounds and compositions, as described in this invention. Preferably, after daily administration of the drug for 5 days, the tumor volume is reduced by at least about 25%. Preferably, after daily administration of the drug for 5 days, the tumor volume is reduced by at least about 50%. Preferably, after daily administration of the drug for 5 days, the tumor volume is reduced by at least about 20-70%. Preferably, after daily administration of the drug for 15 days, the tumor volume is reduced by at least about 25%. Preferably, after daily administration of the drug for 15 days, the tumor volume is reduced by at least about 50%. Preferably, after daily administration of the drug for 15 days, the tumor volume is reduced by at least about 20-70%. Preferably, after daily administration of the drug for 30 days, the tumor volume is reduced by at least about 25%. Preferably, after daily administration of the drug for 30 days, the tumor volume is reduced by at least about 50%. Preferably after daily administration

- 8,020,624 drugs for 30 days, the tumor volume is reduced by at least about 20-70%.

Also, the proposed polymorphic form can be used to inhibit tumor growth by introducing compounds and compounds, as described in this invention. Preferably, after drug administration, tumor growth is inhibited by at least about 20%. Preferably, after drug administration, tumor growth is inhibited by at least about 40%. Preferably, after drug administration, tumor growth is inhibited by at least about 60%. Preferably, after drug administration, tumor growth is inhibited by at least about 80%. Preferably, after drug administration, tumor growth is inhibited between about 20% and about 100%. Preferably, after drug administration, tumor growth is substantially inhibited.

Preferably, the composition is administered twice daily. Preferably, the composition is administered once a day.

Preferably, the MEK inhibitor does not interfere with the co-administration of another tumor suppressant.

Preferably, the composition is in the form of a tablet, capsule, gelatin capsule, caplet, pellet or granule. Preferably, the composition is in dosage form in the form of a capsule or tablet having a total weight of from about 50 to about 1000 mg. Preferably, the composition is in the form of a capsule or tablet having a total weight selected from the group consisting of 50, 75, 100, 150, 200, 250, 300, 350, 400, 450 and 500 mg. Preferably, the composition is in the form of a capsule or tablet having a total weight of from about 240 mg.

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

Preferably, the composition further comprises at least one disintegrating agent selected from the group consisting of croscarmellose sodium, sodium starch glycolate, crospovidone, methyl cellulose, alginic acid, sodium alginate, starch derivatives, concrete and wigum.

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

Preferably, the composition further comprises at least one wetting agent or surfactant selected from the group consisting of sodium lauryl sulfate, glycerol, sorbitan oleates, sorbitan stearates, polyoxyethylene sorbitan laurate, palmitate, stearate, oleate or hexaolate, polyoxyethylene stearyl alcohol and.

The compositions can be used in the form of a capsule or tablet, and the capsule or tablet releases at least 60% of the drug in 30 minutes using Pharmacopoeia I8 (I8P) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as medium dissolution. Preferably, the composition is in the form of a capsule or tablet, and the capsule or tablet releases about 60-100% of the drug in 30 minutes using Pharmacopoeia I8 (I8P) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as a dissolution medium. Preferably, the composition is in the form of a capsule or tablet, and the capsule or tablet releases about 60-90% of the drug in 30 minutes using Pharmacopoeia I8 (I8P) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as a dissolution medium. Preferably, the composition is in the form of a capsule or tablet, and the capsule or tablet releases about 6080% of the drug in 30 minutes using Pharmacopoeia I8 (I8P) Apparatus II at 50 rpm with 1% sodium lauryl sulfate in water as a dissolution medium.

A batch of capsules or tablets, each containing from about 1 to about 50 mg of the compound, as described in this invention, has an acceptance number according to IPR relative to the uniformity of the composition is less than about 15.

Inhibition of MEK enzyme

The proposed polymorphic form can be used to inhibit the MEK enzyme. Preferably, the enzyme is inhibited by at least about 1%. Preferably, the enzyme is inhibited by at least about 2%. Preferably, the enzyme is inhibited by at least about 3%. Preferably, the enzyme is inhibited by at least about 4%. Preferably, the enzyme is inhibited by at least about 5%. Preferably, the enzyme is inhibited by at least about 10%. Preferably, the enzyme is inhibited by at least about 20%. Preferably, the enzyme is inhibited by at least about 25%. Preferably, the enzyme is inhibited by at least about 30%. Preferably, the enzyme is inhibited by at least about 40%.

- 9,020,624

Preferably, the enzyme is inhibited by at least about 50%. Preferably, the enzyme is inhibited by at least about 60%. Preferably, the enzyme is inhibited by at least about 70%. Preferably, the enzyme is inhibited by at least about 75%. Preferably, the enzyme is inhibited by at least about 80%. Preferably, the enzyme is inhibited by at least about 90%. Preferably, the enzyme is substantially completely inhibited. Preferably, the MEK enzyme is a MEK kinase. Preferably, the MEK enzyme is MEK1. Preferably, the MEK enzyme is MEK2. Preferably, contacting occurs in the cell. Preferably, the cell is a mammalian cell. Preferably, the mammalian cell is a human cell.

MEK-mediated violation

The proposed polymorphic form can be used to treat a disorder mediated by MEK in an individual suffering from said disorder by administering to said individual an effective amount of a composition that contains the active compound. Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the pharmaceutical composition is in a form suitable for oral administration. Preferably, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release preparation, solution, suspension, for parenteral injection in the form of a sterile solution, suspension or emulsion, for topical administration as an ointment or cream, or for rectal administration as a suppository . Preferably, the pharmaceutical composition is in unit dosage forms suitable for single administration of exact doses. Preferably, the pharmaceutical composition further comprises a pharmaceutical carrier, excipient and / or adjuvant.

Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.01 to about 7 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably, the amount of active compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. In further or additional embodiments, the compound of formula I is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day. Preferably, the individual suffering from a disorder mediated by MEK is a mammal. Preferably, the individual is a human.

Preferably, the composition containing the active compound is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, a composition that contains an active compound is administered in combination with at least one other therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the MEK-mediated disorder is selected from the group consisting of inflammatory diseases, infections, autoimmune disorders, stroke, ischemia, cardiac abnormalities, neurological disorders, fibrogenetic disorders, proliferative disorders, hyperproliferative disorders, non-malignant hyperproliferative disorders, tumors, leukemia, neoplasms, malignant neoplasms, carcinomas, metabolic diseases, malignant disease, vascular restenosis, psoriasis, atherosclerosis, evmatoidny arthritis, osteoarthritis, heart failure, chronic pain, neuropathic pain, dry eye closure glaucoma and open angle glaucoma. Preferably, the MEK-mediated disorder is an inflammatory disease. Preferably, a violation mediated by MEK,

- 10,020,624 is a hyperproliferative disease. Preferably, the MEK-mediated disorder is selected from the group consisting of tumors, leukemia, neoplasms, malignant neoplasms, carcinomas, and malignant disease. Preferably, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, stomach cancer, kidney cancer, colon and rectal cancer, or leukemia. Preferably, the fibrogenetic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid masses, interstitial nephritis, or pulmonary fibrosis.

Effects Provided

The proposed polymorphic form can provide an effect in a patient, where the effect is selected from the group including the inhibition of various malignant neoplasms, immunological diseases and inflammatory diseases. Preferably, the effect is the inhibition of various malignant neoplasms. Preferably, the effect is an inhibition of immunological diseases. Preferably, the effect is inhibition of inflammatory diseases.

Preferably, the active compound composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the active compound composition is administered in combination with at least one other therapeutic agent.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.01 to about 7 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably, the amount of active compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. Preferably, the active compound is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day. Preferably, the individual suffering from a malignant neoplasm is a mammal. Preferably, the individual is a human.

The proposed polymorphic form can be used to destroy, inhibit the growth or destruction of a cancer cell by contacting said cell with such an amount of a composition that is effective for destroying, inhibiting the growth or destruction of said cancer cell.

Preferably, the cancer cells include cells of the brain, breast, lung, ovary, pancreas, prostate, kidney, or colon and rectum. Preferably, the composition is administered with at least one other therapeutic agent. Preferably, the therapeutic agent is taxol, botresomib, or both. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agents are selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the cancer cells are destroyed. Preferably, 1% of the cancer cells are destroyed. Preferably, 2% of the cancer cells are destroyed. Preferably, 3% of the cancer cells are destroyed. Preferably, 4% of the cancer cells are destroyed. Preferably, 5% of the cancer cells are destroyed. Preferably, 10% of the cancer cells are destroyed. Preferably, 20% of the cancer cells are destroyed. Preferably 25% of the cancer cells are destroyed. Preferably, 30% of the cancer cells are destroyed. Preferably, 40% of the cancer cells are destroyed. Preferably, 50% of the cancer cells are destroyed. Preferably, 60% of the cancer cells are destroyed. Preferably, 70% of the cancer cells are destroyed. Preferably, 75% of the cancer cells are destroyed. Preferably, 80% of the cancer cells are destroyed. Preferably, 90% of the cancer cells are destroyed. Preferably, 100% of the cancer cells are destroyed. Preferably, 11 020624 but essentially all cancer cells are destroyed. Preferably, the above destruction occurs within one day, five days, ten days, one month, two months, six months or one year.

Preferably, the cancer cells are destroyed. Preferably 1% of the cancer cells are destroyed. Preferably, 2% of the cancer cells are destroyed. Preferably, 3% of the cancer cells are destroyed. Preferably, 4% of the cancer cells are destroyed. Preferably, 5% of the cancer cells are destroyed. Preferably, 10% of the cancer cells are destroyed. Preferably, 20% of the cancer cells are destroyed. Preferably, 25% of the cancer cells are destroyed. Preferably, 30% of the cancer cells are destroyed. Preferably, 40% of the cancer cells are destroyed. Preferably, 50% of the cancer cells are destroyed. Preferably, 60% of the cancer cells are destroyed. Preferably, 70% of the cancer cells are destroyed. Preferably, 75% of the cancer cells are destroyed. Preferably, 80% of the cancer cells are destroyed. Preferably, 90% of the cancer cells are destroyed. Preferably, 100% of the cancer cells are destroyed. Preferably, substantially all cancer cells are destroyed. Preferably, the above killing of malignant cells occurs within one day, five days, ten days, one month, two months, six months or one year.

Preferably, the growth of cancer cells is inhibited. Preferably, the growth of cancer cells is inhibited by approximately 1%. Preferably, the growth of cancer cells is inhibited by approximately 2%. Preferably, the growth of cancer cells is inhibited by approximately 3%. Preferably, the growth of cancer cells is inhibited by approximately 4%. Preferably, the growth of cancer cells is inhibited by approximately 5%. Preferably, the growth of cancer cells is inhibited by approximately 10%. Preferably, the growth of cancer cells is inhibited by approximately 20%. Preferably, the growth of cancer cells is inhibited by approximately 25%. Preferably, the growth of cancer cells is inhibited by approximately 30%. Preferably, the growth of cancer cells is inhibited by approximately 40%. Preferably, the growth of cancer cells is inhibited by approximately 50%. Preferably, the growth of cancer cells is inhibited by approximately 60%. Preferably, the growth of cancer cells is inhibited by approximately 70%. Preferably, the growth of cancer cells is inhibited by approximately 75%. Preferably, the growth of cancer cells is inhibited by approximately 80%. Preferably, the growth of cancer cells is inhibited by approximately 90%. Preferably, the growth of cancer cells is inhibited by approximately 100%. Preferably, the above inhibition occurs within one day, five days, ten days, one month, two months, six months or one year.

The proposed polymorphic form can be used to reduce the size of the tumor, inhibit the increase in tumor size, reduce tumor proliferation, or prevent tumor proliferation in an individual by administering to said individual an effective amount of a composition that contains the active compound. Preferably, the size of the tumor is reduced. Preferably, the tumor size is reduced by at least 1%. Preferably, the tumor size is reduced by at least 2%. Preferably, the tumor size is reduced by at least 3%. Preferably, the tumor size is reduced by at least 4%. Preferably, the tumor size is reduced by at least 5%. Preferably, the tumor size is reduced by at least 10%. Preferably, the tumor size is reduced by at least 20%. Preferably, the tumor size is reduced by at least 25%. Preferably, the tumor size is reduced by at least 30%. Preferably, the tumor size is reduced by at least 40%. Preferably, the tumor size is reduced by at least 50%. Preferably, the tumor size is reduced by at least 60%. Preferably, the tumor size is reduced by at least 70%. Preferably, the tumor size is reduced by at least 75%. Preferably, the tumor size is reduced by at least 80%. Preferably, the tumor size is reduced by at least 85%. Preferably, the tumor size is reduced by at least 90%. Preferably, the tumor size is reduced by at least 95%. Preferably, the tumor is eradicated. Preferably, the tumor size does not increase. Preferably, the above effects on tumor size occur within one day, five days, ten days, one month, two months, six months or one year.

Preferably, tumor proliferation is reduced. Preferably, tumor proliferation is reduced by at least 1%. Preferably, tumor proliferation is reduced by at least 2%. Preferably, tumor proliferation is reduced by at least 3%. Preferably, tumor proliferation is reduced by at least 4%. Preferably, tumor proliferation is reduced by at least 5%. Preferably, tumor proliferation is reduced by at least 10%. Preferably, tumor proliferation is reduced by at least 20%. Preferably, tumor proliferation is reduced by at least 25%. Preferably, tumor proliferation is reduced by at least 30%. Preferably, tumor proliferation is reduced by at least 40%. Preferably, tumor proliferation is reduced by at least 50%. Preferably, tumor proliferation is reduced by at least 60%. Preferably, tumor proliferation is reduced by at least 70%. Preferably, tumor proliferation is reduced by at least 75%. Preferably Tumor Proliferation

- 12,020,624 is reduced by at least 75%. Preferably, tumor proliferation is reduced by at least 80%. Preferably, tumor proliferation is reduced by at least 90%. Preferably, tumor proliferation is reduced by at least 95%. Preferably, tumor proliferation is prevented. Preferably, the above effects on cell proliferation occur within one day, five days, ten days, one month, two months, six months, or one year.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the composition is administered in combination with at least one other therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.01 to about 7 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably, the amount of active compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. Preferably, the active compound is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day. Preferably, the individual suffering from a malignant neoplasm is a mammal. Preferably, the individual is a human.

Proliferative diseases

The proposed polymorphic form can be used to treat or prevent a proliferative disease in an individual by administering to said individual an effective amount of a composition that contains the active compound. Preferably, the proliferative disease is a malignant neoplasm, psoriasis, restenosis, an autoimmune disease, or atherosclerosis. Preferably, the proliferative disease is a hyperproliferative disease. Preferably, the proliferative disease is selected from the group consisting of tumors, leukemia, neoplasms, malignant neoplasms, carcinomas and malignant disease. Preferably, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, colon and rectal cancer, stomach cancer, head and neck cancer, or leukemia. Preferably, the fibrogenetic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, liver cirrhosis, keloid formation, interstitial nephritis or pulmonary fibrosis, preferably stomach cancer, brain cancer, breast cancer, lung cancer, non-small cell lung cancer, ovarian cancer, cancer pancreas, liver cancer, prostate cancer, kidney cancer, colorectal cancer, or leukemia. Preferably, the cancer is brain cancer or cancer of the adrenal cortex. In further or additional embodiments, the implementation of the malignant neoplasm is breast cancer. Preferably, the cancer is ovarian cancer. Preferably, the cancer is pancreatic cancer. Preferably, the cancer is prostate cancer. Preferably, the cancer is kidney cancer. Preferably, the malignant neoplasm is colorectal cancer. Preferably, the cancer is myeloid leukemia. Preferably, the cancer is glioblastoma. Preferably, the malignant neoplasm is follicular lymphoma. Preferably, the malignant neoplasm 13 020624 is acute leukemia of B cell precursors. Preferably, the malignant neoplasm is a B-cell chronic lymphocytic leukemia. Preferably, the malignant neoplasm is mesothelioma. Preferably, the malignant neoplasm is small cell lung cancer. Preferably, the cancer is gastric cancer.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the composition is administered in combination with at least one other therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors.

Preferably, the therapeutic agent is selected from taxol, botresomib, or both. Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally.

Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.01 to about 7 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably, the amount of active compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. Preferably, the active compound is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day. Preferably, the individual suffering from a proliferative disease is a mammal. Preferably, the individual is a human.

Inflammatory diseases

The proposed polymorphic form can be used to treat or prevent an inflammatory disease in an individual by administering to said individual an effective amount of a composition that contains the active compound. Preferably the inflammatory disease is selected from the group consisting of chronic inflammatory diseases, rheumatoid arthritis, rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, Still's disease, acute rheumatoid arthritis, enteropathic arthritis, neuropathic arthritis, psoriatic arthritis, purulent arthritis, atherosclerosis, systemic lupus erythematosus, inflammatory bowel disease, irritable bowel syndrome, ulcerative colitis, reflux esophagitis, Crohn’s disease, gastritis, asthma, allergies, respiratory distress syndrome, pancreatitis, chronic obstructive pulmonary disease, pulmonary fibrosis, psoriasis, eczema or scleroderma.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the composition is administered in combination with at least one other therapeutic agent.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.01 to about 7 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably, the amount of active compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

- 14,020,624

Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. Preferably, the active compound is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day. Preferably, the individual suffering from an inflammatory disease is a mammal. Preferably, the individual is a human.

Malignant neoplasm

The proposed polymorphic form can be used to treat or prevent malignant neoplasms in an individual by administering to said individual an effective amount of a composition that contains the active compound. Preferably, the cancer is brain cancer, breast cancer, stomach cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, colon and rectal cancer, or leukemia. Preferably, the fibrogenetic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid masses, interstitial nephritis, or pulmonary fibrosis. Preferably, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, stomach cancer, colon and rectal cancer, or leukemia. Preferably, the cancer is brain cancer or cancer of the adrenal cortex. Preferably, the cancer is breast cancer. Preferably, the cancer is ovarian cancer. Preferably, the cancer is pancreatic cancer.

Preferably, the cancer is prostate cancer.

Preferably, the cancer is kidney cancer. Preferably, the malignant neoplasm is colorectal cancer. Preferably, the cancer is myeloid leukemia. Preferably, the cancer is glioblastoma. Preferably, the malignant neoplasm is follicular lymphoma. Preferably, the cancer is acute leukemia of B cell precursors. Preferably, the malignant neoplasm is a B-cell chronic lymphocytic leukemia. Preferably, the malignant neoplasm is mesothelioma. Preferably, the malignant neoplasm is small cell lung cancer. Preferably, the cancer is gastric cancer.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the composition is administered in combination with at least one other therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of active compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of active compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of active compound is from about 0.001 to about 7 g / day. Preferably, the amount of active compound is from about 0.01 to about 7 g / day. Preferably, the amount of active compound is from about 0.02 to about 5 g / day. Preferably, the amount of active compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of active compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient.

Preferably, dosage levels above the upper limit of the above range may be necessary. Preferably, the active compound is administered in a single dose, once a day. Preferably, the active compound is administered in multiple doses, more than once a day. Preferably, the active compound is administered twice daily. Preferably, the active compound is administered three times a day. Preferably, the active compound is administered four times a day. Preferably, the active compound is administered more than four times per day. Preferably, the individual suffering from a malignant neoplasm is a mammal. Preferably

- 15,020,624 an individual is a human being.

Inclusion by reference

All publications and patent applications referred to in this patent are incorporated into the present invention by reference in the same way, if each individually specified publication or patent application was specifically and individually incorporated by reference.

Brief Description of the Figures

Characteristic features of the invention are set forth in more detail in the appended claims. A better understanding of the features and advantages of the present invention is provided by reference to the following detailed description, which sets forth illustrative embodiments of the invention in which the principles of the invention and their accompanying figures are used.

In FIG. Figure 1 shows graphs of the average tumor volume over time (days) in mice with implanted A375 melanoma cells, Colo205 colorectal tumors, A431 epidermoid tumors, or HT-29 colorectal tumors. Mice were dosed orally (25, 50, or 100 mg / kg) once a day for 14 days.

In FIG. Figure 2 shows a graph of% inhibition of tumor growth (% TC1) in A375 xenograft mice that were given 50 mg / kg дозы drug doses. 25 mg / kg VGO, 50 mg / kg ΟΌ and 12.5 mg / kg VGO.

In FIG. Figure 3 shows a plasma concentration graph (1ode nM) versus pECK% inhibition in female pi / pi mice with implanted Co1O205 tumor cells. Mice were given a single dose of 2.5, 5, 10 or 25 mg / kg.

In FIG. 4 shows a graph of plasma concentration (ng / ml) over time (hours) in people after a single dose of 2 mg (capsules 2x 1 mg), 4 mg (capsules 4x 1 mg) or 6 mg (capsules 6x 1 mg).

In FIG. 5 is a graph of a powder X-ray diffraction pattern (ΡΧΚΌ)-(§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide Form A obtained using a 1pe1 XKS-3000 diffractometer. The graph shows the peak intensity as determined based on the number of pulses per second relative to the diffraction angle of 2 угла in degrees.

In FIG. 6 is a graph of a thermogram of modulated differential scanning calorimetry GO5> C) X- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane -1-sulfonamide Form A, obtained using a TA PMitepe 01000 differential scanning calorimeter. The graph shows the normalized heat flux in units of W / gram (W / g) relative to the measured sample temperature in ° C.

In FIG. Figure 7 shows graphs of the pattern ΡΧΚΌ ΡΧΚΌ- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide form A ( above) and Ν- (δ) - (3,4difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1sulfonamide amorphous (below), obtained with using a 1pe1 XKS-3000 diffractometer. The graph shows the peak intensity as determined based on the number of pulses per second relative to the diffraction angle of 2 угла in degrees.

In FIG. Figure 8 shows the sorption / desorption isotherm of dynamic evaporation (Όνδ) Ν- (δ) - (3,4difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane- 1 sulfonamide Form A, obtained using an νΤΙ 8CA-100 evaporative sorption analyzer.

In FIG. Figure 9 shows a thermogram of thermogravimetry (TS) ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide form ( A) obtained using a thermogravimetric analyzer TA 1pigitep1 2950.

In FIG. 10 (A) and 10 (B) show the growth arrest of dividing A375 cells in the Lod phase exposed to elevated concentrations of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) - 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide. Cells were analyzed for ATP content. 100% growth arrest was determined using 1 μM ^ (§) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide .

In FIG. Figure 11 shows a 48-hour AK study in A375 cells. The dividing A375 cells in the Lb phase were exposed to ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide and ΡΌ-325901 for 48 hours and were analyzed for AK release.

In FIG. 12 (A) -12 (C) shows growth inhibition using ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (A) human colorectal carcinoma cells So1o205 (C1 ₅₀ = 11 nM); (B) A375 cells (C1 ₅₀ = 22 nM); and (C) inhibition of MOA-MB231 cells that do not exhibit induced ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide growth arrest in 2-dimensional studies dependent on anchoring.

In FIG. 13 (A) shows the inhibition of cell growth of human colorectal carcinoma Co1O205, with values of C1 ₅₀ at a concentration of 6 and 11 nm, respectively. In FIG. 13 (B) shows inhibition

- 16,020,624 growth of A375 cells with O1 values of ₅₀ at a concentration of 5 and 22 nM.

In FIG. 14 (A) and 14 (B) show the effect of L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane- 1-sulfonamide on the implementation of the cell cycle, indicating that the effect on A375 cells L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide causes a stop in phase 01 of the cell cycle, as indicated by cell depletion in both phase 02 and phase 8.

In FIG. 15 (A) and 15 (B) show the effect of L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane- 1-sulfonamide per stomach cancer cell line A08 (gastric adenocarcinoma) after 3 days (Fig. 15 (A)) and 6 days (Fig. 15B). On the оси axis, the number of cells relative to the filler is shown, and on the X axis in μM, the concentration of L- (8) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3 -dihydroxypropyl) cyclopropane-1-sulfonamide.

In FIG. 16 shows the average liver weight in mice bearing the tumor after treatment with Y- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2 , 3-dihydroxypropyl) cyclopropane-1-sulfonamide (2 mg / kg, once daily, orally; 10 mg / kg, once daily, oral and 50 mg / kg, once daily, oral).

In FIG. 17 shows the weight of the liver tumor in mice bearing the tumor after treatment with Ν (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3 -dihydroxypropyl) cyclopropane-1sulfonamide (2 mg / kg, once a day, orally; 10 mg / kg, once a day, orally and 50 mg / kg, once a day, orally).

In FIG. 18 shows the average tumor weight after treatment with ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (2; 10 and 50 mg / kg).

In FIG. Figure 19 shows the inhibition of Ηδ746ΐ cell proliferation graphically for the number of cells (relative to the vehicle) as a function of the concentration ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2, 3-dihydroxypropyl) cyclopropane-1-sulfonamide.

In FIG. 20 (A) is a graph comparing the corresponding levels of apoptosis at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl ) cyclopropane-1-sulfonamide on day 5 of treatment of MU522 cells of non-small cell lung cancer (Ν8ΟΕΟ). In FIG. 20 (B) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of H358 cells of non-small cell lung cancer (Ν8ΟΕΟ) sSC. FIG. 20 (C) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 6 of treatment of A549 non-small cell lung cancer cells (Ν8ΟΕΟ). In FIG. 20 (Ό) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of H727 non-small cell lung cancer cells (Ν8ΟΕΟ). In FIG. 20 (E) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of the treatment of HT29 colon cells. In FIG. 20 (P) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 6 of treatment of colon cells HCT116. In FIG. 20 (0) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of hepatoma cells of the colon NIN7. In FIG. 20 (H) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of I2-O8 sarcoma cells. In FIG. 20 (1) is a graph showing the corresponding apoptosis levels at elevated concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on day 5 of treatment of glioma cells Ό37.

In FIG. 21 shows the selectivity of compound A to MEK1 and MEK2 relative to a panel of 205 enzymes at 10 μM. Cell lines were Co1o205, A375, A431 and NT-29.

In FIG. 22 is a graph showing the increase in paw volume in each of the treated groups and the decrease in edema relative to the vehicle control after administration of 6, 20, 60 and 200 mg / kg doses to rats ^ (8) - (3,4-difluoro-2- ( 2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide in rat carrageenan-induced paw edema.

In FIG. 23 (A) shows the inhibition of swelling in the acute phase arthritis model of adjuvant in rats treated with 2, 6 and 20 mg / kg ^ (8) - (3,4-difluoro-2- (2-fluoro- 4 iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide. In FIG.

- 17,020,624

23 (C) shows the inhibition of swelling in an adjuvant-induced arthritis model in the delayed phase in rats treated with 2, 6 and 20 mg / kg L- (B) - (3,4-difluoro-2- (2-fluorine) -4-iodophenylamino) 6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropan-1-sulfonamide.

In FIG. 24 shows the average arthritis score for arthritis induced by the anti-collagen antibody (CA1A) in mice treated with 1, 3, and 10 mg / kg ΟΌ Y- (B) - (3,4-difluoro-2- (2- fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

DETAILED DESCRIPTION OF THE INVENTION

The section headings used in this invention are presented for organizational purposes only and should not be construed as limiting the described scope of the invention. All documents or portions of documents cited in a patent, including, without limitation, patents, patent applications, articles, books, manuals and textbooks, are hereby incorporated by reference in their entirety for any purpose.

Specific chemical terminology

Unless specifically indicated otherwise, all technical and scientific terms used in the present invention have the same meanings as are commonly understood by one skilled in the art to which the claimed subject matter relates. All patents, patent applications, published materials that are referred to in the present invention, unless expressly indicated otherwise, are fully incorporated by reference. In the event that there are many definitions for the terms used in this invention, the value indicated in this section has the advantage. If reference is made to the IRE or other such identifier or address, it is understood that such identifiers may change and specific information on the Internet can be added and deleted, but using the search you can find equivalent information on the Internet or other suitable reference material. For this purpose, the link indicates the availability and open dissemination of such information.

It is intended that the foregoing general description and the following detailed description be illustrative and illustrative only and not limit any claimed objects. In the present invention, the use of the singular encompasses the plural, unless expressly indicated otherwise. It should be noted that, as used in the description and claims, the singular include reference to the plural, unless the context clearly dictates otherwise. It should also be noted that the use of either denotes and / or, unless specifically indicated otherwise. In addition, the use of the term including, as well as other forms, such as includes, include and included, are not limiting.

Definitions of standard chemical terms can be found in reference books, including Sageuii Bipyogd Ayuapsey Ogdashe Szet181tu, 4th ed., Volumes A (2000) and B (2001), P1epit PTE88, No. \ ν Wogk. Unless specifically indicated otherwise, conventional methods of mass spectroscopy, NMR, HPLC, IR, and UV and visible spectroscopy and pharmacology are used, which are known to those skilled in the art. If specific definitions are not provided, then the nomenclature used, laboratory methods and techniques of analytical chemistry, synthetic organic chemistry and medical and pharmaceutical chemistry, as described in this invention, are known in the art. For chemical syntheses, chemical analyzes, pharmaceuticals, drug preparation, and patient delivery and treatment, standard techniques can be used. Interactions and purification procedures can be carried out, for example, using sets of manufacturer's specifications or as is usually carried out in the art or as described in this invention. The following techniques and procedures are usually carried out according to conventional methods well known in the art and as described in various general and more specific manuals that are cited and discussed in the present invention. For the entire description, the groups and their substituents may be selected by one of ordinary skill in the art to provide stable parts and compounds.

Specific pharmaceutical terminology

The term MEK inhibitor, as used in this invention, refers to a compound that exhibits 1C ₅₀ with respect to MEK activity of not more than about 100 μM or not more than about 50 μM, as determined in the study of Mek1 kinase, generally described in this invention . 1C ₅₀ represents the concentration of the inhibitor, which reduces the activity of the enzyme (eg, MEK) by half from the maximum level. It was found that the compounds disclosed in this invention exhibit an inhibitory effect on the activity of MEK. The compounds of the present invention preferably exhibit 1C ₅₀ with respect to MEK of not more than about 10 μM, more preferably not more than about 5 μM, even more preferably not more than about 1 μM, and most preferably not more than about 200 nM, as defined in Mek1 study described in this invention.

The term subject, patient or individual, as used in this invention, refers to individuals suffering from a disorder, etc., and encompasses mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class

- 18,020,624 mammals: humans, primates other than humans, such as chimpanzees, and other anthropoid apes and species of monkeys; farm animals such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs and cats; laboratory animals, including rodents, such as rats, mice and guinea pigs, etc. Examples of non-mammals are, but are not limited to, birds, fish, etc. In one embodiment of the methods and compositions provided in the present invention, the mammal is a human.

The terms treat, treatment or treatment and other grammatical equivalents as used in the present invention include alleviating, alleviating or improving the symptoms of a disease or condition, preventing additional symptoms, improving or preventing the metabolic causes underlying the symptoms, inhibiting the disease or condition, for example stopping the development of a disease or condition, alleviating a disease or condition, causing a regression of the disease or condition, alleviating a condition, caused about a disease or condition, or stopping the symptoms of a disease or condition, and also covers prevention. The terms further include providing a therapeutic benefit and / or prophylactic benefit. The therapeutic advantage is understood to mean eliminating or attenuating the underlying disorder being treated. Also, a therapeutic advantage is provided by eliminating or alleviating one or more physiological symptoms associated with the underlying disorder, such that the patient has an improvement, despite the fact that the patient may still suffer from the underlying disease. For the prophylactic benefit of the composition, the patient can be administered at the risk of developing a particular disease or to a patient who exhibits one or more physiological symptoms of the disease, although the disease itself may not yet be diagnosed.

The terms effective amount, a therapeutically effective amount, or a pharmaceutically effective amount, as used in the present invention, refers to the amount of at least one administration agent or compound that is sufficient to treat or prevent a particular disease or condition. The result may be a reduction and / or alleviation of the signs, symptoms or causes of the disease or any other change in the biological system. For example, an effective amount for therapeutic applications is the amount of a composition containing a compound, as described herein, necessary to provide a clinically significant reduction in disease. A suitable effective amount in any particular case can be determined using techniques such as dose-increase studies.

The terms essentially does not contain water and essentially does not contain solvent, as used in the present invention, refers to crystalline polymorphic forms containing less than 0.01, 0.1, 0.2, 0.3, 0.4, 0 , 5, 1 or 2 wt.% Water or solvent, respectively.

The term is essentially the same as used in the present invention, refers to a powder x-ray or differential scanning calorimetry pattern, which may not be identical to that presented in this invention, but which falls within the experimental error, which is taken into account by a specialist in this technical field.

The terms apply, administer, administer, etc., as used in the present invention, refer to methods that can be used to enable delivery of compounds or compositions to a desired site of biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, peritoneal injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), local and rectal administration. Administration methods are known to those skilled in the art that can be used for the compounds and methods described in this invention, for example, as discussed in Tell Aib Obtai, Thé Pagantacoodoxa1 Ba818 Tocarebens, current edition; Regdatop; apb RepipDyup'z. Raptaseibs 1 Zaepsez (current edition), Musk Ribztd Co., Eazlop, Ra. In preferred embodiments, the compounds and compositions described herein are administered orally.

The term acceptable, as used in the present invention, in relation to a preparation, composition or component means the absence of a long-term temporary effect on the general health of the patient being treated.

The term pharmaceutically acceptable, as used in the present invention, refers to a substance, such as a carrier or diluent, that does not alter the biological activity or properties of the compounds as described in this invention, and is relatively non-toxic, i.e. the substance can be administered to an individual without causing unwanted side effects or unwanted interactions with any of the components of the composition in which it is contained.

The term pharmaceutical composition, as used in the present invention, refers to a biologically active compound, optionally mixed with at least one pharmaceutically acceptable chemical component, such as, but not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickeners and / or

- 19,020,624 excipients.

The term carrier, as used in the present invention, refers to relatively non-toxic chemical compounds or agents that facilitate the incorporation of the compound into cells or tissues.

The term agonist, as used in the present invention, refers to a molecule, such as a compound, drug, enzyme activator or hormone modulator, which enhances the activity of another molecule or the activity of the receptor site.

The term antagonist, as used in the present invention, refers to a molecule, such as a compound, drug, enzyme inhibitor or hormone modulator, which reduces or prevents the action of another molecule or the activity of the receptor site.

The term modulate, as used in the present invention, refers to the interaction with the target either directly or indirectly in order to change the activity of the target, including, by way of example only, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target or increasing the activity of the target.

The term modulator, as used in the present invention, refers to a molecule that interacts with a target either directly or indirectly. Interactions include, but are not limited to, interactions between an agonist and an antagonist.

The terms enhancing or enhancing, as used in the present invention, means increasing or prolonging the effectiveness or duration of the desired effect. Therefore, with regard to the effect of therapeutic agents, the term enhancing refers to the ability to increase or prolong either the effectiveness or duration of action of other therapeutic agents on the system. A reinforcing effective amount, as used in the present invention, refers to an amount sufficient to enhance the effect of another therapeutic agent in the desired system.

The terms pharmaceutical combination, administration of an additional treatment, administration of an additional therapeutic agent and the like, as used in the present invention, refers to pharmaceutical treatment resulting from mixing or combining several active components and includes both fixed and uncommitted combinations of active components. The term fixed combination means that at least one of the compounds described in this invention and at least one co-administered agent are both simultaneously administered to the patient in the form of a single unit or dose. The term “non-fixed combination” means that at least one of the compounds described in this invention and at least one co-administered agent are administered to the patient as separate units simultaneously, together or separately with variable time intervals, where such administration provides effective levels two or more compounds in the patient’s body. This also applies to cocktail therapies, for example the administration of three or more active ingredients.

The terms co-administration, administration in combination with and their grammatical equivalents or the like, as used in the present invention, encompasses the administration of the selected therapeutic agents to one patient and encompasses treatment regimens according to which the agents are administered in the same or different routes of administration either simultaneously or various time intervals. In some embodiments, the compounds described in this invention will be co-administered with other agents. This term encompasses the administration of two or more agents to an animal in such a way that both agents and / or their metabolites are present in the animal at the same time. These include simultaneous administration in separate compositions, administration at different time intervals in individual compositions, and / or administration in compositions in which both agents are present. Thus, in some embodiments, the compounds of the invention and the other (s) agent (a) are administered in the same composition. In some embodiments, the compounds of the invention and the other agent (s) are mixed in a composition.

Polymorphs include various crystalline packaged groupings of compounds of identical elemental composition. Polymorphs can have various X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability and solubility. Various factors, such as solvent recrystallization, crystallization rate, and storage temperature, can cause a single crystalline form to predominate.

- 20,020,624

The crystalline polymorphic form of I- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

The invention relates to crystalline polymorphic form A And- (§) - (3,4-difluoro-2- (2-fluoro-4iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

which has a specific x-ray powder diffraction pattern. In some embodiments, the X-ray powder diffraction pattern comprises at least about 50% of the peaks shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least about 70% of the peaks shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least about 90% of the peaks shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern is substantially the same as the X-ray powder diffraction pattern shown in FIG. 5.

The invention also relates to crystalline polymorphic form A And- (§) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide , which exhibits a specific picture of differential scanning calorimetry. In some embodiments, the specific differential scanning calorimetry pattern is substantially identical to the differential scanning calorimetry pattern shown in FIG. 6. In some embodiments, the crystalline polymorphic form A has a melting point that appears, as determined by differential scanning calorimetry, at about 143 ° C.

The invention also relates to the polymorphic form of I- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, which obtained using a method including the crystallization step of amorphous I- (§) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1- sulfonamide from a solvent. The polymorphic form of I- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide can be obtained using a method comprising the crystallization step of amorphous I- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide from a mixture hexane and ethyl acetate.

The invention also relates to pharmaceutical compositions that contain an effective amount of crystalline polymorphic form A And- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3- dihydroxypropyl) cyclopropane-1-sulfonamide; and a pharmaceutically acceptable carrier or excipient. Other aspects of the invention relate to a pharmaceutical composition that comprises crystalline polymorphic form A and at least one excipient or carrier.

Crystalline polymorphic form A And- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide is suitable for the treatment of or prevent malignant neoplasms or inflammatory diseases. The invention further relates to the use of crystalline polymorphic form A for the treatment or prevention of malignant neoplasms or inflammatory diseases. Other aspects of the invention relate to the use of crystalline polymorphic form A for the treatment or prevention of proliferative disease.

In some embodiments, the composition comprises at least about 50% of a compound that exhibits a x-ray powder diffraction comprising at least 50% of the peaks identified in the x-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 70% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 90% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern is substantially identical to the X-ray powder diffraction pattern shown in FIG. 5.

In some embodiments, the composition comprises at least about 75% of a compound that exhibits a powder x-ray containing at least 50% of the peaks identified in the powder x-ray shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 70% of the peaks identified in the 21,206,624 X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 90% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern is substantially identical to the X-ray powder diffraction pattern shown in FIG. 5.

In some embodiments, the composition comprises at least about 90% of a compound that exhibits a x-ray powder diffraction comprising at least 50% of the peaks identified in the x-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 70% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 90% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern is substantially identical to the X-ray powder diffraction pattern shown in FIG. 5.

In some embodiments, substantially all of the compounds in the composition exhibit an X-ray powder diffraction pattern comprising at least 50% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 70% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern comprises at least 90% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. In some embodiments, the X-ray powder diffraction pattern is substantially identical to the X-ray powder diffraction pattern shown in FIG. 5.

In some embodiments, the crystalline polymorph present in the composition has a melting point that appears, as determined by differential scanning calorimetry, at about 143 ° C. In some embodiments, the implementation of the crystalline polymorph is essentially free of water. In some embodiments, the implementation of the crystalline polymorph is essentially free of solvent.

In some embodiments, the composition comprises at least about 50% of a compound that exhibits a differential scanning calorimetry pattern substantially identical to the differential scanning calorimetry pattern shown in FIG. 6. In some embodiments, the implementation of the crystalline polymorph has a melting point, which is manifested, as determined using differential scanning calorimetry, at about 143 ° C. In some embodiments, the implementation of the crystalline polymorph is essentially free of water. In some embodiments, the implementation of the crystalline polymorph is essentially free of solvent.

In some embodiments, the composition comprises at least about 75% of a compound that exhibits a differential scanning calorimetry pattern substantially identical to the differential scanning calorimetry pattern shown in FIG. 6. In some embodiments, the implementation of the crystalline polymorph has a melting point, which is manifested, as determined using differential scanning calorimetry, at about 143 ° C. In some embodiments, the implementation of the crystalline polymorph is essentially free of water. In some embodiments, the implementation of the crystalline polymorph is essentially free of solvent.

In some embodiments, the composition comprises at least about 90% of a compound that exhibits a differential scanning calorimetry pattern substantially identical to the differential scanning calorimetry pattern shown in FIG. 6. In some embodiments, the implementation of the crystalline polymorph has a melting point, which is manifested, as determined using differential scanning calorimetry, at about 143 ° C. In some embodiments, the implementation of the crystalline polymorph is essentially free of water. In some embodiments, the implementation of the crystalline polymorph is essentially free of solvent.

In some embodiments, substantially all of the compounds in the composition exhibit a differential scanning calorimetry pattern substantially identical to the differential scanning calorimetry pattern shown in FIG. 6. In some embodiments, the implementation of the crystalline polymorph has a melting point, which is manifested, as determined using differential scanning calorimetry, at about 143 ° C. In some embodiments, the implementation of the crystalline polymorph is essentially free of water. In some embodiments, the implementation of the crystalline polymorph is essentially free of solvent.

In some embodiments, the polymorphic form of Y- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide is prepared using a process comprising a crystallization step of amorphous L- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide. Preferably, the 2220624 crystallization step involves crystallization from a mixture of ethyl acetate and heptane, for example, a mixture of ethyl acetate and heptane contains from about 1-4 parts of ethyl acetate to about 2-10 parts of heptane or more specifically, from about 2 parts of ethyl acetate up to about 5 parts of heptane.

Preferably, the compound is formulated as a medicament for the rapid release of the compound. Preferably, the compound is formulated as a medicament for the sustained release of the compound. Preferably, the compound is formulated as a medicament for sustained release of the compound.

In some embodiments, the composition is presented in unit dosage form in the form of a tablet. Preferably, the composition is in capsule dosage form. The composition can be converted into a dosage form in the form of a capsule or tablet, and many other alternative compositions and methods of preparation can be used. REFERENCES: (1) Vepiidum, Thy δοίepse and Prgase, O Pagtasu, 20th ed., 2000, (2) Pbagtaseijs1 Bokade Egtk Tablets volumes 1-3, 1989 and (3) Mobegp Pagtaseysa18 4th ed., 2002. Various methods of preparation can be used, including dry mixing, wet granulation, roll pressing, extrusion, spheronization, coating, and spray drying. Soft gels and methods for their preparation are also possible.

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

In some embodiments, the composition comprises a disintegrant. Preferably, the disintegrating agent is selected from croscarmellose sodium, sodium starch glycolate, crospovidone, methyl cellulose, alginic acid, sodium alginate, starch derivatives, concrete and wigum. Preferably, the disintegrating agent is croscarmellose sodium.

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

In some embodiments, the composition comprises a wetting agent or surfactant. Preferably, the wetting agent or surfactant is selected from sodium lauryl sulfate, glycerol, sorbitan oleates, sorbitan stearates, polyoxyethylene sorbitan laurate, palmitate, stearate, oleate or hexaolate, polyoxyethylene stearyl alcohol and sorbitan monolaurate. Preferably, the wetting agent or surfactant is sodium lauryl sulfate.

Additional excipients, such as glidants, flavors and colorants, can also be added. Additional active excipients can be found in Thb Nabbok about аг аг аг се се се се 1 1 ар ар ар г ей з,,, 5th ed., 2005 and the database of inactive components of EBA (Commission on Drug and Nutrient Control).

Also provided in the present invention are pharmaceutical compositions that contain an effective amount of crystalline polymorphic form A - (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3 -dihydroxypropyl) cyclopropane-1-sulfonamide. In some embodiments, the pharmaceutical compositions comprise an effective amount of crystalline polymorphic form A-(§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions are for treating disorders. In some embodiments, the pharmaceutical compositions are for treating disorders in a mammal. In some embodiments, the pharmaceutical compositions are for treating disorders in humans. In some embodiments, the pharmaceutical compositions are for treating or preventing inflammatory diseases. In some embodiments, the pharmaceutical compositions are for the treatment or prophylaxis of proliferative diseases.

Methods of Using Compounds and Compositions

The proposed polymorphic form can be used to provide an effect in a patient who is selected from the group including the inhibition of various malignant neoplasms, immunological diseases and inflammatory diseases. Preferably, the compound is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the effect is the inhibition of various malignant neoplasms. Preferably, the effect is an inhibition of immunological diseases. Preferably, the effect is inhibition of inflammatory

- 23,020,624 diseases.

Any compositions described and claimed in the present invention can be used in the methods disclosed in this section.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy or surgery, or any combination thereof. Preferably, the composition is administered in combination with at least one other therapeutic agent.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of the compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of the compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of the compound is from about 0.001 to about 7 g / day. Preferably, the amount of the compound is from about 0.01 to about 7 g / day. Preferably, the amount of the compound is from about 0.02 to about 5 g / day. Preferably, the amount of the compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of the compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

In some embodiments of the compositions and methods described herein, MEK protein kinase inhibitors are provided, further comprising polymorph (A), which is present in an amount of from about 0.1 to about 200 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of from about 0.2 to about 100 mg. In other embodiments, the MEK protein kinase inhibitor comprises a polymorph (A), which is present in an amount of from about 0.3 to about 90 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of from about 0.4 to about 80 mg. In other embodiments, the MEK protein kinase inhibitor comprises a polymorph (A), which is present in an amount of from about 0.5 to about 70 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of from about 0.4 to about 80 mg. In other embodiments, the MEK protein kinase inhibitor comprises a polymorph (A), which is present in an amount of from about 0.5 to about 70 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of from about 1 to about 60 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of about 1.5 to about 50 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of from about 2 to about 45 mg. In other embodiments, the MEK protein kinase inhibitor comprises polymorph (A), which is present in an amount of from about 2.5 to about 40 mg.

In some embodiments of the compositions and methods described herein, MEK protein kinase inhibitors are provided, further comprising polymorph (A), which is present in an amount of about 0.1 mg, about 0.2 mg, about 0.25 mg, about 0, 3 mg, approximately 0.4 mg, approximately 0.5 mg, approximately 0.6 mg, approximately 0.7 mg, approximately 0.8 mg, approximately 0.9 mg, approximately 1 mg, approximately 1.5 mg, approximately 2 mg, approximately 2.5 mg, approximately 3 mg, approximately 3, 5 mg, approximately 4.0 mg, approximately 4.5 mg, approximately 5 mg, approximately 5.5 mg, approximately 6 mg, approximately 6.5 mg, approximately 7 mg, approximately 7.5 mg, approximately 8 mg, approximately 8.5 mg, approximately 9 mg, approximately 9.5 mg, approximately 10 mg, approximately 10.5 mg, approximately 11 mg, approximately 11.5 mg, approximately 12 mg, approximately 12.5 mg and / or approximately 13 mg approximately 14 mg or approximately 15 mg.

In some embodiments of the compositions and methods described herein, MEK protein kinase inhibitors are provided, further comprising polymorph (A), which is present in an amount of about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, approximately 45 mg, approximately 50 mg, approximately 55 mg, approximately 60 mg, approximately 65 mg, approximately 75 mg, approximately 80 mg, approximately 85 mg, approximately 90 mg, approximately 95 mg, p approximately 100 mg, approximately 110 mg, approximately 120 mg, approximately 125 mg, approximately 130 mg, approximately 140 mg, approximately 150 mg, approximately 160 mg, approximately 170 mg, approximately 175 mg, approximately 180 mg, approximately 190 mg, or approximately 200 mg

Preferably, the compound is administered in a single dose, once a day. Preferably, the compound is administered in multiple doses, more than once a day. Preferably, the compound is administered twice daily. Preferably, the compound is administered three times a day. Preferably, the compound is administered four times a day. Preferably, the compound is administered more than four times per day. Preferably, the individual suffering from a malignant neoplasm is a mammal. Preferably, the individual is a human.

Diseases and disorders that are modulated by MEK

Also provided in this invention are methods for modulating MEK activity by contacting MEK with an amount of compound sufficient to modulate MEK activity. Modulation may be inhibition or activation of MEK activity. Preferably, inhibition of MEK activity is provided by contacting the MEK with an amount of compound sufficient to inhibit MEK activity. Preferably, inhibition of MEK activity in the solution is provided by contacting said solution with an amount of a compound sufficient to inhibit MEK activity in said solution. Preferably, inhibition of MEK activity in a cell is provided by contacting said cell with an amount of a compound as described herein sufficient to inhibit MEK activity in said cell. Preferably, inhibition of MEK activity in tissue is provided by contacting said tissue with an amount of a compound as described herein sufficient to inhibit MEK activity in said tissue. Preferably, inhibition of MEK activity in an organism is provided by contacting said organism with an amount of a compound as described herein sufficient to inhibit MEK activity in said organism. Preferably, inhibition of MEK activity in an animal is provided by contacting said animal with an amount of a compound as described herein sufficient to inhibit MEK activity in said animal. Preferably, inhibition of MEK activity in a mammal is provided by contacting said mammal with an amount of a compound as described herein sufficient to inhibit MEK activity in said mammal. Preferably, inhibition of MEK activity in a person is provided by contacting said person with an amount of a compound as described herein sufficient to inhibit MEK activity in said person.

Compounds and compositions can modulate the activity of MEK enzymes; and as such are suitable for the treatment of diseases or conditions in which aberrant MEK activity contributes to the pathology and / or symptoms of the disease or condition.

Compounds and compositions can be used to treat a disorder or condition that is modulated by the MEK cascade in a mammal, including humans, by administering to said mammal an amount of a compound effective to modulate said cascade. A suitable dose for a particular patient can be determined according to known methods by those skilled in the art.

The compounds and compositions can be used to inhibit the MEK enzyme, which includes contacting said MEK enzyme with an amount of a composition that contains a compound sufficient to inhibit said enzyme in which said enzyme is inhibited. Preferably, the enzyme is inhibited by at least about 1%. Preferably, the enzyme is inhibited by at least about 2%. Preferably, the enzyme is inhibited by at least about 3%. Preferably, the enzyme is inhibited by at least about 4%. Preferably, the enzyme is inhibited by at least about 5%. Preferably, the enzyme is inhibited by at least about 10%. Preferably, the enzyme is inhibited by at least about 20%. Preferably, the enzyme is inhibited by at least about 25%. Preferably, the enzyme is inhibited by at least about 30%. Preferably, the enzyme is inhibited by at least about 40%. Preferably, the enzyme is inhibited by at least about 50%. Preferably, the enzyme is inhibited by at least about 60%. Preferably, the enzyme is inhibited by at least about 70%. Preferably, the enzyme is inhibited by at least about 75%. Preferably, the enzyme is inhibited by at least about 80%. Preferably, the enzyme is inhibited by at least about 90%. Preferably, the enzyme is substantially completely inhibited. Preferably, the MEK enzyme is a MEK kinase. Preferably, the MEK enzyme is MEK1. Preferably, the MEK enzyme is MEK2. Preferably, contacting occurs in the cell. Preferably, the cell is a mammalian cell. Preferably, the mammalian cell is a human cell.

Compounds and compositions can be used to treat a disorder mediated by MEK in an individual suffering from said disorder by administering to said individual an effective amount of a composition that contains the compound. Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the pharmaceutical com position 2520624 is in a form suitable for oral administration. Preferably, the pharmaceutical composition is in the form of a tablet, capsule, pill, powder, sustained release preparation, solution, suspension, for parenteral injection in the form of a sterile solution, suspension or emulsion, for topical administration as an ointment or cream, or for rectal administration as a suppository . Preferably, the pharmaceutical composition is in unit dosage forms suitable for single administration of exact doses. Preferably, the pharmaceutical composition further comprises a pharmaceutical carrier, excipient and / or adjuvant.

Preferably, the amount of the compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of the compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of the compound is from about 0.001 to about 7 g / day. Preferably, the amount of the compound is from about 0.01 to about 7 g / day. Preferably, the amount of the compound is from about 0.02 to about 5 g / day. Preferably, the amount of the compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of the compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the compound is administered in a single dose, once a day. Preferably, the compound is administered in multiple doses, more than once a day. Preferably, the compound is administered twice daily. Preferably, the compound is administered three times a day. Preferably, the compound is administered four times a day. Preferably, the compound is administered more than four times per day. Preferably, the individual suffering from a disorder mediated by MEK is a mammal. Preferably, the individual is a human.

Preferably, the composition containing the compound is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the composition that contains the compound is administered in combination with at least one other therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the MEK-mediated disorder is selected from the group consisting of inflammatory diseases, infections, autoimmune disorders, stroke, ischemia, cardiac abnormalities, neurological disorders, fibrogenetic disorders, proliferative disorders, hyperproliferative disorders, non-malignant hyperproliferative disorders, tumors, leukemia, neoplasms, malignant neoplasms, carcinomas, metabolic diseases, malignant disease, vascular restenosis, psoriasis, atherosclerosis, evmatoidny arthritis, osteoarthritis, heart failure, chronic pain, neuropathic pain, dry eye closure glaucoma and open angle glaucoma. Preferably, the MEK-mediated disorder is an inflammatory disease. Preferably, the MEK-mediated disorder is a hyperproliferative disease. Preferably, the MEK-mediated disorder is selected from the group consisting of tumors, leukemia, neoplasms, malignant neoplasms, carcinomas and malignant diseases such as stomach cancer, brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer glands, kidney cancer, colorectal cancer, or leukemia. Preferably, the fibrogenetic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid masses, interstitial nephritis, or pulmonary fibrosis.

Malignant neoplasm

Also provided in this invention are compounds, pharmaceutical compositions and methods for treating, preventing or preventing malignant neoplasm in an individual, which comprises administering to said individual an effective amount of the compound. Preferably, the compound is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, colon and rectal cancer, stomach cancer or leukemia. Preferably, the fibrogenetic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid masses, interstitial nephritis, or pulmonary fibrosis. Preferably, the cancer is cancer

- 26,020,624 brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, colorectal cancer, leukemia, melanoma, thyroid cancer, or basal cell cancer. Preferably, the cancer is brain cancer or cancer of the adrenal cortex. Preferably, the cancer is breast cancer. Preferably, the cancer is ovarian cancer. Preferably, the cancer is pancreatic cancer. Preferably, the cancer is prostate cancer. Preferably, the cancer is kidney cancer. Preferably, the malignant neoplasm is colorectal cancer. Preferably, the cancer is myeloid leukemia. Preferably, the cancer is glioblastoma. Preferably, the malignant neoplasm is follicular lymphoma. Preferably, the cancer is acute leukemia of B cell precursors. Preferably, the malignant neoplasm is a B-cell chronic lymphocytic leukemia. Preferably, the malignant neoplasm is mesothelioma. Preferably, the malignant neoplasm is small cell lung cancer. Preferably, the cancer is gastric cancer.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the composition is administered in combination with at least one therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the amount of the compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of the compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of the compound is from about 0.001 to about 7 g / day. Preferably, the amount of the compound is from about 0.01 to about 7 g / day. Preferably, the amount of the compound is from about 0.02 to about 5 g / day. Preferably, the amount of the compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of the compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the compound is administered in a single dose, once a day. Preferably, the compound is administered in multiple doses, more than once a day. Preferably, the compound is administered twice daily. Preferably, the compound is administered three times a day. Preferably, the compound is administered four times a day. Preferably, the compound is administered more than four times per day. Preferably, the individual suffering from a malignant neoplasm is a mammal. Preferably, the individual is a human.

Abnormal cell growth

Also provided in this invention are compounds, pharmaceutical compositions and methods for inhibiting abnormal cell growth. Preferably, abnormal cell growth occurs in a mammal. Methods for inhibiting abnormal cell growth include administering an effective amount of a compound wherein abnormal cell growth is inhibited. Methods of inhibiting abnormal cell growth in a mammal include administering to the mammal an amount of a compound, where amounts of the compound are effective in inhibiting abnormal cell growth in a mammal.

Preferably, the methods include administering an effective amount of a compound in combination with an amount of a chemotherapeutic agent, wherein the amounts of the compound and chemotherapeutic agent are jointly effective in inhibiting abnormal cell growth. Currently, many chemotherapeutic agents are known in the art and can be used in combination with the compound of the invention. Preferably, the chemotherapeutic agent is selected from the group consisting of mitosis inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors and antiandrogens.

- 27,020,624

The present invention also provides methods for inhibiting abnormal cell growth in a mammal, which include administering to the mammal an amount of a compound in combination with radiation therapy, where the amounts of the compound used in combination with radiation therapy are effective in inhibiting abnormal cell growth or treating hyperproliferative disorder in a mammal. Radiation therapy techniques are known in the art, and these techniques can be used in combination therapy as described in this invention. The administration of a compound in this combination therapy can be determined as described in this invention.

The present invention also describes methods and pharmaceutical compositions for inhibiting abnormal cell growth in a mammal, which comprises an amount of a compound and an amount of one or more substances selected from anti-angiogenesis substances, signal transduction inhibitors and antiproliferative agents.

Anti-angiogenesis agents such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors and COX-11 inhibitors (cyclooxygenase 11) can be used in combination with the compound of the present invention and pharmaceutical compositions as described in this invention. Examples of suitable COX-P inhibitors include SEEVKEX ™ (alecoxib), valdecoxib, and rofecoxib. Examples of suitable matrix metalloproteinase inhibitors are described in \ O 96/33172 (publ. October 24, 1996), \ O 96/27583 (publ. March 7, 1996), European patent application EP7304971.1 (filed July 8, 1997 ), European patent application EP99308617.2 (filed October 29, 1999), \ O 98/07697 (publ. February 26, 1998), \\ 'O 98/03516 (publ. January 29, 1998), \ \ 'O 98/34918 (publ. August 13, 1998), \\' O 98/34915 (publ. August 13, 1998), \ O 98/33768 (publ. August 6, 1998), \ O 98/30566 (publ. July 16, 1998), EP606046 (publ. July 13, 1994), EP931788 (publ. July 28, 1999), \ O 90/05719 (publ. May 31, 1990), \ O 99/52910 (published on October 21, 1999), \ O 99/52889 (o Rev. October 21, 1999), \ O 99/29667 (published June 17, 1999), international application PCT / 1B98 / 01113 (filed July 21, 1998), European patent application EP99302232.1 (filed March 25) 1999), patent application OV9912961.1 (filed June 3, 1999), provisional patent application I860 / 148464 (filed August 12, 1999), patent I85863949 (issued July 26, 1999), patent I85861510 (issued January 19, 1999) and EP780386 (publ. June 25, 1997), all of these sources are fully incorporated into this invention by reference. Some MMP-2 and MMP-9 inhibitors have little or no inhibitory effect on MMP-1 activity, while some selectively inhibit MMP-2 and / or AMP-9 with respect to other matrix metalloproteinases (i.e., MAP-1 , MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12 and MMP-13). Some specific examples of MMP inhibitors useful in the present invention are AO-3340, KO 323555, and KZ 13-0830.

The present invention also describes a method of destroying, inhibiting the growth or destruction of a cancer cell, which comprises contacting said cell with an amount of composition effective to destroy, inhibit the growth or destruction of said cancer cell. Preferably, the cancer cells include cells of the brain, breast, lung, ovary, pancreas, prostate, kidney, or colon and rectum.

Preferably, the composition is administered with at least one therapeutic agent. Preferably, the therapeutic agent is taxol, botresomib, or both. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agents are selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors.

Preferably, the cancer cells are destroyed. Preferably, 1% of the cancer cells are destroyed. Preferably, 2% of the cancer cells are destroyed. Preferably 3% of the cancer cells are destroyed. Preferably 4% of the cancer cells are destroyed. Preferably, 5% of the cancer cells are destroyed. Preferably 10% of the cancer cells are destroyed. Preferably, 20% of the cancer cells are destroyed. Preferably 25% of the cancer cells are destroyed. Preferably, 30% of the cancer cells are destroyed. Preferably, 40% of the cancer cells are destroyed. Preferably, 50% of the cancer cells are destroyed. Preferably, 60% of the cancer cells are destroyed. Preferably, 70% of the cancer cells are destroyed. Preferably, 75% of the cancer cells are destroyed. Preferably, 80% of the cancer cells are destroyed. Preferably, 90% of the cancer cells are destroyed. Preferably, 100% of the cancer cells are destroyed. Preferably, substantially all cancer cells are destroyed.

Preferably, the cancer cells are destroyed. Preferably 1% of the cancer cells are destroyed. Preferably, 2% of the cancer cells are destroyed. Preferably 3% of the cancer cells are destroyed. Preferably 4% of the cancer cells are destroyed. Preferably, 5% of the cancer cells are destroyed. Preferably 10% of the cancer cells are destroyed. Preferably, 20% of the cancer cells are destroyed. Preferably 25% of the cancer cells are destroyed. Preferably, 30% of the cancer cells are destroyed. 28 020624 Preferably, 40% of the cancer cells are destroyed. Preferably, 50% of the cancer cells are destroyed. Preferably, 60% of the cancer cells are destroyed. Preferably, 70% of the cancer cells are destroyed. Preferably, 75% of the cancer cells are destroyed. Preferably, 80% of the cancer cells are destroyed. Preferably, 90% of the cancer cells are destroyed. Preferably, 100% of the cancer cells are destroyed. Preferably, substantially all cancer cells are destroyed.

Preferably, the growth of cancer cells is inhibited. Preferably, the growth of cancer cells is inhibited by approximately 1%. Preferably, the growth of cancer cells is inhibited by approximately 2%. Preferably, the growth of cancer cells is inhibited by approximately 3%. Preferably, the growth of cancer cells is inhibited by approximately 4%. Preferably, the growth of cancer cells is inhibited by approximately 5%. Preferably, the growth of cancer cells is inhibited by approximately 10%. Preferably, the growth of cancer cells is inhibited by approximately 20%. Preferably, the growth of cancer cells is inhibited by approximately 25%. Preferably, the growth of cancer cells is inhibited by approximately 30%. Preferably, the growth of cancer cells is inhibited by approximately 40%. Preferably, the growth of cancer cells is inhibited by approximately 50%. Preferably, the growth of cancer cells is inhibited by approximately 60%. Preferably, the growth of cancer cells is inhibited by approximately 70%. Preferably, the growth of cancer cells is inhibited by approximately 75%. Preferably, the growth of cancer cells is inhibited by approximately 80%. Preferably, the growth of cancer cells is inhibited by approximately 90%. Preferably, the growth of cancer cells is inhibited by approximately 100%.

Also provided herein are methods for inhibiting abnormal cell growth. Preferably, abnormal cell growth occurs in a mammal. Methods for inhibiting abnormal cell growth include administering an effective amount of crystalline polymorphic form A Ν (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl ) cyclopropane-1-sulfonamide, where abnormal cell growth is inhibited. Methods of inhibiting abnormal cell growth in a mammal include administering to the mammal an amount of crystalline polymorphic form A ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl ) cyclopropane-1-sulfonamide, where the amount of crystalline polymorphic form A ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane -1-sulfonamide is effective for inhibiting abnormal cell growth in a mammal.

Preferably, the methods include administering an effective amount of crystalline polymorphic form A No. (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1- sulfonamide in combination with the amount of chemotherapeutic agent, where the amounts of crystalline polymorphic form A ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide and a chemotherapeutic agent are jointly effective in inhibiting abnormal cell growth.

Various chemotherapeutic agents are currently known in the art and can be used in combination with the compounds and compositions of the invention. Preferably, the chemotherapeutic agent is selected from the group consisting of mitosis inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors and antiandrogens.

Preferred methods for inhibiting abnormal cell growth in a mammal include administering to the mammal an amount of crystalline polymorphic form A ^ (8) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3- dihydroxypropyl) cyclopropane-1-sulfonamide in combination with radiation therapy, where the amount of crystalline polymorphic form A Ν- (δ) - (3,4difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- ( 2,3-dihydroxypropyl) cyclopropane-1sulfonamide in combination with radiation therapy is effective for inhibiting abnormal cell growth. Radiation therapy techniques are known in the art, and these techniques can be used in combination therapy as described in this invention.

Treatment of hyperproliferative disorder

The present invention describes a method of treating a hyperproliferative disorder in a mammal, including a human, which comprises administering to said mammal a therapeutically effective amount of a compound.

Preferably, the compound is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the proliferative disease is a malignant neoplasm, psoriasis, restenosis, an autoimmune disease, or atherosclerosis. Preferably, the proliferative disease is a hyperproliferative disease. Preferably, the proliferative disease is selected from the group consisting of tumors, leukemia, neoplasms, malignant neoplasms, carcinomas and malignant disease. Preferably, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, stomach cancer, colorectal cancer, or leukemia. Preferably, the fibrogenetic disorder is scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, cirrhosis, keloid masses, interstitial nephritis, or pulmonary fibrosis. Preferably, the cancer is brain cancer, breast cancer, lung cancer, ovarian cancer, stomach cancer, pancreatic cancer, prostate cancer, kidney cancer, colon and rectal cancer, or leukemia. Preferably, the cancer is brain cancer or cancer of the adrenal cortex. Preferably, the cancer is breast cancer. Preferably, the cancer is ovarian cancer. Preferably, the cancer is pancreatic cancer. Preferably, the cancer is prostate cancer. Preferably, the cancer is kidney cancer. Preferably, the malignant neoplasm is colorectal cancer. Preferably, the cancer is myeloid leukemia. Preferably, the cancer is glioblastoma. Preferably, the malignant neoplasm is follicular lymphoma. Preferably, the cancer is acute leukemia of B cell precursors. Preferably, the malignant neoplasm is an extracellular chronic lymphocytic leukemia. Preferably, the malignant neoplasm is mesothelioma. Preferably, the malignant neoplasm is small cell lung cancer. Preferably, the cancer is gastric cancer.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or a combination thereof. Preferably, a composition that contains a compound of formula I is administered in combination with at least one therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of the compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of the compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of the compound is from about 0.001 to about 7 g / day. Preferably, the amount of the compound is from about 0.01 to about 7 g / day. Preferably, the amount of the compound is from about 0.02 to about 5 g / day. Preferably, the amount of the compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of the compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the compound is administered in a single dose, once a day. Preferably, the compound is administered in multiple doses, more than once a day. Preferably, the compound is administered twice daily. Preferably, the compound is administered three times a day. Preferably, the compound is administered four times a day. Preferably, the compound is administered more than four times per day. Preferably, the individual suffering from a proliferative disease is a mammal. Preferably, the individual is a human.

Tumor size

The present invention describes a method for reducing tumor size, inhibiting an increase in tumor size, reducing tumor proliferation, or preventing tumor proliferation in an individual, which comprises administering to said individual an effective amount of a compound. Preferably, the compound is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the size of the tumor is reduced. Preferably, the tumor size is reduced by at least 1%. Preferably, the tumor size is reduced by at least 2%. Preferably, the tumor size is reduced by at least 3%. Preferably, the tumor size is reduced by at least 4%. Preferably, the tumor size is reduced by at least 5%. Preferably, the tumor size is reduced by at least 10%. Preferably, the tumor size is reduced by at least 20%. Preferably, the tumor size is reduced by at least 25%. Preferably, the size of the tumor is reduced by at least 30%. Preferably, the tumor size is reduced by at least 40%. Preferably, the tumor size is reduced by at least 50%. Preferably, the tumor size is reduced by at least 60%. Preferably, the tumor size is reduced by at least 70%. Preferably, the tumor size is reduced by at least 75%. Preferably, the tumor size is reduced by at least 80%. Preferably, the tumor size is reduced by at least 85%. Preferably, the tumor size is reduced by at least 90%. Preferably, the tumor size is reduced by at least 95%. Preferably, the tumor is eradicated. Preferably, the tumor size does not increase.

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

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the adjunctive therapy is radiation therapy, chemotherapy, surgery, or any combination thereof. Preferably, the composition is administered in combination with at least one therapeutic agent. Preferably, the therapeutic agent is selected from the group consisting of cytotoxic agents, anti-angiogenesis agents, and antitumor agents. Preferably, the antitumor agent is selected from the group consisting of alkylating agents, antimetabolites, epidophyllotoxins; antitumor enzymes, topoisomerase inhibitors, procarbazines, mitoxantrons, platinum coordination complexes, biological response modifiers and growth inhibitors, hormonal / antihormonal therapeutic agents and hematopoietic growth factors. Preferably, the therapeutic agent is selected from taxol, botresomib, or both.

Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of the compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of the compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of the compound is from about 0.001 to about 7 g / day. Preferably, the amount of the compound is from about 0.01 to about 7 g / day. Preferably, the amount of the compound is from about 0.02 to about 5 g / day. Preferably, the amount of the compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of the compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the compound is administered in a single dose, once a day. Preferably, the compound is administered in multiple doses, more than once a day. Preferably, the compound is administered twice daily. Preferably, the compound is administered three times a day. Preferably, the compound is administered four times a day. Preferably, the compound is administered more than four times per day. Preferably, the individual suffering from a malignant neoplasm is a mammal. Preferably, the individual is a human.

Inflammatory disease

The present invention describes a method of treating, preventing or preventing an inflammatory disease in an individual, which comprises administering to said individual an effective amount of a compound. Preferably, the compound is administered as a component of a composition that further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the inflammatory disease is selected from the group consisting of chronic inflammatory diseases, rheumatoid arthritis, rheumatoid arthritis, spondylarthropathy, ankylosing spondylitis, gout, tendonitis, bursitis, sciatica, gouty arthritis, osteoarthritis, Still's disease, acute rheumatoid arthritis, rheumatoid rheumatoid arthritis, rheumatoid arteritis arthritis, pus - 31 020624 arthritis, atherosclerosis, systemic lupus erythematosus, inflammatory bowel disease, irritable bowel syndrome, carried specific ulcerative colitis, reflux esophagitis, Crohn’s disease, gastritis, asthma, allergies, respiratory distress syndrome, pancreatitis, chronic obstructive pulmonary disease, pulmonary fibrosis, psoriasis, eczema or scleroderma.

Preferably, the composition is administered in combination with adjunctive therapy. Preferably, the composition is administered in combination with at least one therapeutic agent. Preferably, the composition is administered orally, intraduodenally, parenterally (including intravenously, subcutaneously, intramuscularly, intravascularly or by infusion), topically or rectally. Preferably, the amount of the compound is in the range of from about 0.001 to about 1000 mg / kg body weight / day. Preferably, the amount of the compound is in the range of from about 0.5 to about 50 mg / kg / day. Preferably, the amount of the compound is from about 0.001 to about 7 g / day. Preferably, the amount of the compound is from about 0.01 to about 7 g / day. Preferably, the amount of the compound is from about 0.02 to about 5 g / day. Preferably, the amount of the compound is from about 0.05 to about 2.5 g / day. Preferably, the amount of the compound is from about 0.1 to about 1 g / day. Preferably, dosage levels below the lower limit of the above range may be more than sufficient. Preferably, dosage levels above the upper limit of the above range may be necessary.

Preferably, the compound is administered in a single dose, once a day. Preferably, the compound is administered in multiple doses, more than once a day. Preferably, the compound is administered twice daily. Preferably, the compound is administered three times a day. Preferably, the compound is administered four times a day. Preferably, the compound is administered more than four times per day. Preferably, the individual suffering from an inflammatory disease is a mammal. Preferably, the individual is a human.

Ways of doing

The compounds and compositions described herein can be administered either singly or in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition according to standard pharmaceutical practice.

The invention also describes pharmaceutical compositions that contain crystalline polymorph ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane- 1-sulfonamide (Form A). The compounds and compositions described herein can be administered either singly or in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition according to standard pharmaceutical practice. The introduction can be carried out using any method that is able to deliver compounds to the site of action. These methods include, but are not limited to, enteral delivery (including an oral, gastric, or duodenal nasal tube, rectal suppository, and rectal enema), parenteral routes (injection or infusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, vitreous, epidural and subcutaneous), inhalation, transdermal, transmucosis nogo, sublingual, buccal and topical (including cutaneous, dermal, enema, eye drops, ear drops, intranasal, vaginal) administration although the most suitable route may depend upon for example the condition of the patient and disorders. Administration methods that can be used with the compounds and methods of the invention will be known to those skilled in the art. By way of example only, the compounds described in this invention can be administered topically to a site in need of treatment, for example, by local infusion during surgery, topical administration such as creams or ointments, injections, a catheter or implant, the implant is made for example, from a porous, non-porous or gelatinous substance, including membranes, such as sialastic membranes or fibers. The introduction can also be carried out by direct injection into the area of diseased tissue or organ.

The administration of the compounds and compositions described in this invention can be accomplished using any method that is capable of delivering the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), local and rectal administration. For example, the compounds described in this invention can be administered locally to the area in need of treatment. This can be achieved using, for example, but not limited to, local infusion during surgery, local injection, for example, cream, ointment, injection, catheter or implant, the implant is made, for example, of a porous, non-porous or gelatinous substance, including membranes such as sialastic membranes or fibers. The introduction can also be carried out by direct injection into the site (or the previous site) of the tumor or neoplasm or precancerous tissue. 32,020,624 are known to those skilled in the art for preparations and administration methods that can be used for the compounds and methods of the invention, for example, as discussed in Ooooytap apy Oytap, Thieu Ryagtaso1od1sa1 Ba818 o £ Tyegareiysk, current edition; Regdatop; and KettDyupK Ryagtaseijsa 1 Biepsek (current edition), Musk RybKypd Co., EaChop, Ra.

Medicines are suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intraarterial and intramedullary), intraperitoneal, transmucosal, transdermal, rectal and local (including skin, trans buccal, sublingual and intraocular) administration may depend on, although the most appropriate , for example, from a condition and disorder in a patient. Medicines are suitably presented in unit dosage form and can be prepared using any of the methods well known in the pharmaceutical art. All methods include the step of contacting a compound of the present invention or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate or derivative (active component) thereof with a carrier, which consists of one or more additional components. In general, drugs are prepared by uniformly and intimately contacting the active component with liquid carriers or finely divided solid carriers or both of these components, and then, if necessary, forming the product into the desired drug.

Medicines suitable for oral administration may be in the form of discrete components, such as capsules, starch capsules or tablets, each of which contains a predetermined amount of the active component; in the form of powder or granules; in the form of a solution or suspension in an aqueous liquid or non-aqueous liquid; or as an oil-inlet liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be in the form of a bolus, electuary or paste.

Medicines that are suitable for oral administration include tablets, tight-fitting capsules made of gelatin, and soft, sealed capsules made of gelatin and an emollient such as glycerin or sorbitol. Tablets may be prepared by compression or molding, optionally with one or more accessory components. Compressed tablets can be prepared by compressing in a suitable machine the active component in a free-flowing form, such as powder or granules, optionally mixed with binders, inert diluents or lubricants, surface-active or dispersing agents. Molded tablets may be prepared by forming in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. Tablets may optionally be coated or corrugated and may be formulated so as to provide slow or controlled release of the active ingredient. All drugs for oral administration must be in dosage forms suitable for such administration. Tightly placed capsules or tablets may contain the active ingredient; in admixture with a filler such as microcrystalline cellulose, silicate microcrystalline cellulose, gelatinized starch, lactose, calcium diphosphate or compressible sugar; a binder such as hypromellose, povidone or starch paste; a disintegrating agent such as croscarmellose sodium, crospovidone or sodium starch glycolate; a surfactant such as sodium lauryl sulfate and / or lubricants and process modifiers such as talc, magnesium stearate, stearic acid or colloidal silicon dioxide and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycols. In addition, you can add stabilizers. Dragee cores are coated with suitable coatings. For this, concentrated sugar solutions are used, which optionally may contain gum arabic, talc, polyvinylpyrrolidone, carbopol, polyethylene glycol and / or titanium dioxide, varnish solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments can be added to tablets and dragee coatings to identify or characterize various combinations of doses of the active compounds.

Medicines can be prepared for parenteral administration by injection, for example, by bolus injection or continuous infusion. Injectable preparations may be in unit dosage form, for example, in ampoules or in containers for repeated use, to which a preservative is added. The compositions may be presented in such forms as suspensions, solutions or emulsions in oily or aqueous fillers, and may contain prescription agents, such as suspending, stabilizing and / or dispersing agents. The preparations may be in single-use or multi-use containers, for example, sealed ampoules and vials, and may be stored in powder form or in a freeze-dried (lyophilized) state, to which it is only necessary to add a sterile liquid carrier, for example, saline or sterile pyrogen-free water, immediately before use. Extemporaneous solutions and suspensions for injection can be prepared from sterile powders, granules and tablets of the above varieties.

- 33,020,624

Preparations for parenteral administration include aqueous and non-aqueous (oily) sterile injectable solutions of the active compounds, which may contain antioxidants, buffers, bacteriostatic agents and dissolved substances that render the drug isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickeners. Suitable lipophilic solvents or excipients are fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or substances that increase the solubility of the compounds, which makes it possible to prepare highly concentrated solutions.

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

For buccal or sublingual administration, the compositions may be prepared in the form of tablets, lozenges, lozenges, or gel preparations prepared in a conventional manner. Such compositions may contain the active component in a flavored base such as sucrose and gum arabic or tragacanth gum.

Pharmaceutical preparations may also be formulated as rectal compositions, such as suppositories or retention enemas, for example, containing conventional suppository bases, such as cocoa butter, polyethylene glycol or other glycerides.

Pharmaceutical preparations may be administered topically, which is an unsystematic administration. This includes applying the compounds of the present invention externally to the epidermis or the oral cavity and instilling such a compound into the eye, ear and nose so that the compound does not substantially enter the bloodstream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

Pharmaceutical preparations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation, such as gels, liniment, lotions, creams, ointments or pastes, and drops suitable for administration to the eyes, ears or nose. The active component may contain for local administration from 0.001 to 10 wt.%, For example from 1 to 2 wt.% Of the drug. However, it may contain up to 10 wt.% Or may contain less than 5 wt.% Or from 0.1 to 1 wt.% Of the drug.

Pharmaceutical formulations for administration by inhalation are suitably delivered from a powder blower, a nebulizer of pressurized packages, or other suitable means for delivering an aerosol spray. Packages under pressure may contain a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined using a valve to deliver the metered amount. Alternatively, for administration by inhalation or insufflation, the pharmaceutical preparations may be in the form of an anhydrous powder composition, for example, a powder mixture of a compound and a suitable base of powder, such as lactose or starch. The powder composition may be in unit dosage form, for example, in capsules, cartridges, gelatin or blister packs, from which the powder can be administered using an inhaler or powder blower.

It is also understood that, in addition to the components preferably described above, the compounds and compositions described in this invention may include other agents generally accepted in the art, depending on the type of preparation, for example, preparations for oral administration may include flavorings.

Compositions

It should be noted that any compositions and compounds described in this invention can be used in any of the compositions discussed in this section, which are not limiting and should not be construed in this way.

The compounds or compositions described in this invention can be delivered in an excipient, for example a liposome (see, for example, Lapdeg, §1eise 1990, 249, 1527-1533; Tgea1 et al. / -Vegpket and Nb1eg, ed., Ykk, Ν.Υ., pp. 353-365, 1989). The compounds and pharmaceutical compositions described herein may also be delivered in a controlled release system. In one embodiment, a pump can be used (see §eiop, 1987, SCS SA KeG. Vutej. Ups. 14: 201; VisBta1b et al. §Igdegu, 1980 88, 507; §aibek et al. Ν. Upt1. Ί. Meb. 1989, 321 (574)). Additionally, a controlled-release system can be placed close to a therapeutic target (see Soobkop, MeFsa! ArrNsa11opk oG Sop1go11eb Ke1eake,

- 34,020,624

1984, Volume 2, p. 115-138). The pharmaceutical compositions described in this invention may also contain the active ingredient in a form suitable for oral administration, for example in the form of tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups, or elixirs. The compositions are intended for oral use and can be prepared according to any of the methods known in the art for the preparation of pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavors, colorants and preservatives to provide pharmaceutically quality and attractive drugs. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the preparation of tablets. Such excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating substances; fillers such as microcrystalline cellulose, silicate microcrystalline cellulose, gelatinized starch, lactose, calcium diphosphate or compressible sugar; binders such as hypromellose, povidone or starch paste; disintegrating agents such as croscarmellose sodium, crospovidone or sodium starch glycolate; a surfactant such as sodium lauryl sulfate and / or lubricants and process modifiers such as talc, croscarmellose sodium, corn starch or alginic acid; binders, for example starch, gelatin, polyvinylpyrrolidone or gum arabic, and lubricants, for example magnesium stearate, stearic acid or colloidal silicon dioxide and, optionally, talc. The tablets may be uncoated or may be coated according to known techniques in order to obscure the taste of the drug or to slow the decomposition and absorption in the gastrointestinal tract and thus provide a long-lasting effect over a longer period of time. Suitable suitable, for example, are a water-soluble taste masking agent, such as hydroxypropyl methylcellulose or hydroxypropyl cellulose, or a time-delaying substance, such as ethyl cellulose or cellulose acetate. Oral formulations may also be presented in the form of hard gelatin capsules in which the active component is mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin, or in the form of soft gelatin capsules in which the active component is mixed with a water-soluble carrier, such like polyethylene glycol or an oil medium, such as peanut butter, liquid paraffin or olive oil. Dosage forms in the form of capsules and tablets can be prepared using various processing techniques, including dry mixing and wet granulation. In a dry-mixed preparation method, a drug can be incorporated into a dosage form by dry-mixing with excipients, followed by encapsulation in a capsule shell or compression into a tablet form. The dry mixing operation can be carried out in stages and it includes screening stages between the grinding stages to facilitate the formation of a homogeneous mixture. When prepared by wet granulation, the drug can be added to dry excipients and mixed before the addition of a binder, or the drug can be dissolved and added as a solution as part of the granulation. In the wet granulation technique, a surfactant, if used, can be added to dry excipients or added to a binder solution and included as a solution. Capsule dosage forms can also be prepared by dissolving the drug substance in a material with which it can be filled and which is compatible with the shells of hard gelatin capsules, which can then be bonded and sealed. Dosage forms in the form of capsules and tablets can also be obtained by dissolving a drug substance in a material, such as a molten form of high molecular weight polyethylene glycol, and cooling to a solid form, grinding and incorporating this substance into conventional methods for preparing capsules and tablets.

Aqueous suspensions contain the active substance in admixture with excipients suitable for the preparation of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth gum and gum arabic; dispersing or wetting agents may be a naturally occurring phosphatide, for example, lecithin, or condensation products of alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethylene-oxycetanol, or condensation products of non-ethylene oxide, ethylene oxide acids and hexites, such as polyoxyethylene sorbitan monooleate, or products of condensation of ethylene oxide with partial esters, odnymi fatty acids and hexitol anhydrides, for example polietilensorbitmonooleat.

Aqueous suspensions may also contain one or more preservatives, for example ethyl, or npropyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose, saccharin or aspartame.

- 35,020,624

Oily suspensions may be prepared by suspending the active ingredient in a vegetable oil, for example peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweeteners, such as those described above, and flavoring agents may also be added to provide a palatable oral preparation. These compositions are preserved by the addition of an antioxidant such as butylated hydroxyanisole or alpha-tocopherol.

Dispersible powders and granules suitable for the preparation of an aqueous suspension, when water is added, the active component is mixed with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are those agents, examples of which are already indicated above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, for example olive oil or peanut oil, or a mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifiers may include naturally occurring phosphatides, for example soya lecithin, and esters or partial esters, derivatives of fatty acids and hexite anhydrides, such as sorbitan monooleate, and condensation products of said partial ethylene oxide esters, for example polyoxyethylene sorbitol monooleate. Emulsions may also contain sweeteners, flavors, preservatives and antioxidants.

Syrups and elixirs can be prepared with sweeteners, for example glycerin, propylene glycol, sorbitol or sucrose. Such preparations may also contain an irritant, preservative, flavoring and coloring agents and an antioxidant.

The pharmaceutical compositions may be in the form of a sterile injectable aqueous solution. Suitable excipients and carriers may be water, Ringer's solution and isotonic sodium chloride solution. The sterile preparation for injection may also be a sterile oil-in-water microemulsion for injection, in which the active component is dissolved in the oil phase. For example, the active ingredient may first be dissolved in a mixture of soybean oil and lecithin. Then the oil solution is introduced into a mixture of water and glycerol and processed to obtain a microemulsion. Solutions for injection or microemulsions can be introduced into the bloodstream of a patient by local bolus injection. Alternatively, it may be favorable to administer the solution or microemulsion in such a way as to maintain a constant circulating concentration of the compound. To maintain such a constant concentration, a device for continuous intravenous delivery can be used. An example of such a device is the Es11cc SAEERI8 ™ Model 5400 Intravenous Pump. The pharmaceutical compositions may be in the form of sterile aqueous or oily injection suspensions for intramuscular and subcutaneous administration. Such a suspension can be prepared according to methods known in the art using suitable dispersing or wetting agents and suspending agents as described above. The sterile injectable preparation may also be a sterile solution for injection or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Additionally, sterile, fatty oils are also generally acceptable as a solvent or suspending medium. To do this, you can use any soft fatty oil, including synthetic mono- or diglycerides. Additionally, fatty acids such as oleic acid can be used to prepare injection compositions.

Pharmaceutical compositions may also be administered in the form of suppositories for rectal administration of a drug. These compositions can be prepared by mixing the inhibitors with a suitable non-irritating excipient, which is solid at ordinary temperature but liquid at rectal temperature, and therefore will melt in the rectum, releasing the drug. Such materials are cocoa butter, gelatin, which contains glycerin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and esters of fatty acids and polyethylene glycol.

For topical use, creams, ointments, gels, solutions or suspensions, etc. containing the compound or composition of the present invention are suitable for topical administration. As used in the present invention, topical administration may include mouthwashes and throats.

The pharmaceutical compositions can be administered in intranasal form by topical application of suitable intranasal excipients and delivery devices, or by transdermal route using such forms of transdermal patches as are well known to one of ordinary skill in the art.

The preparations can usually be in dosage form and can be prepared using any of the methods well known in the pharmaceutical field. All methods include the step of contacting a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug or solvate (active component) thereof with a carrier, which consists of one or more accessory components. In general, preparations are prepared by uniformly and closely contacting the active component with liquid carriers or finely divided solid carriers or both types of carriers, and then, if necessary, forming the product into the desired preparation. Methods for preparing various pharmaceutical compositions with a specific amount of active compound are known, or will be apparent to those skilled in the art. For administration in the form of a transdermal delivery, the dosage form must obviously be continuous, and not periodic, throughout the dosage regimen.

Doses

The amount of pharmaceutical compositions administered will initially depend on the mammal being treated. In cases where pharmaceutical compositions are administered to humans, the daily dose will usually be determined by the attending physician taking into account the dose, which usually varies depending on the age, gender, nutrition, weight, general health and response of a particular patient, the severity of symptoms in a particular patient indications or condition being treated, severity of the indication or condition being treated, time of administration, route of administration, pharmacokinetics of the composition, rate of excretion, combination of drugs, and and individual opinion of the attending physician. Also, the route of administration may substantially depend on the condition and its severity. The pharmaceutical composition may be presented in unit dosage form. In this form, the drug is divided into doses in one dose containing the appropriate amount of the active component, for example, an effective amount to achieve the desired goal. Determining the appropriate dose for a particular situation is in the skill of a person skilled in the art. In general, treatment begins with lower doses, which are lower for the optimal dose of the compound. After that, the dose is increased by a small amount until the optimal effect is achieved in the given circumstances. For convenience, the total daily dose may be divided and administered in portions throughout the day, if desired. The amount and frequency of administration of the compounds described in this invention and, if appropriate, other therapeutic agents and / or therapies will be adjusted according to the opinion of the attending physician (therapist), taking into account the factors described above. Thus, the amount administered of the pharmaceutical composition may vary significantly. Administration can be carried out in an amount ranging from about 0.001 mg / kg body weight to about 100 mg / kg body weight per day (administered in single or multiple doses), or at least about 0.1 mg / kg body weight per day. A preferred therapeutic dose may include, for example, from about 0.01 to about 7000 mg of the compound, or, for example, from about 0.05 to about 2500 mg. The amount of active compound in a standard dose may vary or be adjusted from about 0.1 to 1000 mg, from about 1 to 300 mg, or 10 to 200 mg, according to the particular application. In some cases, dosing levels below the lower limit of the above range may be more than sufficient, while in some cases higher doses can be applied without causing any harmful side effects, for example, by splitting such large doses into several lower doses for administration throughout the day . The quantity entered will depend on the specific value of Κ. ' ₅₀ used compounds. In combined applications in which the compound is not the only active component, it is possible to administer even lower amounts of the compound and still obtain a therapeutic or prophylactic effect.

Additional dosage is also provided in the present description and claims.

Dosage forms

The pharmaceutical composition may be, for example, in a form suitable for oral administration, such as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection in the form of a sterile solution, suspension or emulsion, for local administration in as an ointment or cream, or for rectal administration as a suppository. The pharmaceutical composition may be in unit dosage forms suitable for single administration of exact doses. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound of the invention as an active ingredient. Additionally, it may include other medical or pharmaceutical agents, carriers, adjuvants, etc.

Examples of forms for parenteral administration are solutions or suspensions of the active compounds in sterile aqueous solutions, for example, aqueous solutions of propylene glycol or dextrose. Such dosage forms may be suitably buffered, if desired.

Suitable pharmaceutical carriers include inert diluents or excipients, water and various organic solvents. Pharmaceutical compositions may contain, if desired, additional components, such as flavoring agents, binders, excipients, etc. Thus, for oral administration, tablets containing various excipients, such as citric acid, may be used in conjunction with various disintegrating agents, such as starch, alginic acid and certain complex silicates and with binders such as sucrose, gelatin and gum arabic. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often used for tabletting. Solid compositions of a similar type can also be used in soft and hard gelatin-filled capsules, including lactose or milk sugar and high molecular weight polyethylene glycols. If aqueous suspensions or elixirs are desired for oral administration, the active compound therein may be combined with various sweeteners or flavorings, coloring agents or colorants and, if desired, emulsifiers or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.

Methods for preparing various pharmaceutical compositions with a specific amount of active compound are known or will be apparent to those skilled in the art. E.g. (1990).

Combination therapy

The crystalline polymorphic form of L- (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (form A) may administered as a single therapy. The crystalline polymorphic form A - (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide can also be introduced into combination with other therapy or therapies.

By way of example only, if hypertension is one of the side effects that a patient experiences when taking the compound described in this invention, then administration of an antihypertensive agent in combination with the compound may be appropriate. Or, by way of example only, the therapeutic efficacy of the compound described in this invention can be enhanced by the administration of an adjuvant (i.e., the adjuvant itself can only have a slight therapeutic advantage, but in combination with another therapeutic agent, the overall therapeutic advantage for patient rises). Or, by way of example only, the advantage that the patient gains can be enhanced by administering the compound described in this invention with another therapeutic agent (which also includes a treatment regimen), which also has a therapeutic advantage. By way of example only, in the treatment of diabetics comprising administering a compound of the invention, an increase in therapeutic benefit can be obtained by administering to a patient another therapeutic agent for diabetics. In any case, regardless of the disease, disorder or condition being treated, the total advantage that the patient receives can only be the additive effect of two therapeutic agents or the patient can get a synergistic advantage.

Other treatments include, but are not limited to, administration of other therapeutic agents, radiation therapy, or both. In cases where the compound described in this invention is administered with other therapeutic agents, the compounds need not be administered in the same composition as other therapeutic agents, and may, due to various physical and chemical characteristics, be administered in a different way. For example, the compounds / compositions may be administered orally to create and obtain a sufficient concentration in the blood, while another therapeutic agent may be administered intravenously. The determination of the route of administration and the appropriateness of administration, if possible, in one pharmaceutical composition, is within the skill of a person skilled in the art. Initial administration can be carried out according to established protocols known in the art, and then, based on the observed actions, the attending physician can modify the dosage, route of administration and time of administration. The preferred choice of the compound (and, if appropriate, another therapeutic agent and / or radiation) will depend on the diagnosis of the attending physician and his understanding of the patient's condition and the appropriate treatment protocol. Other therapeutic agents may include chemotherapeutic agents, such as antitumor agents, for example, selected from the following groups: mitosis inhibitors, such as vinblastine; alkylating agents, for example cisplatin, carboplatin and cyclophosphamide; antimetabolites, for example 5-fluorouracil, cytosine arabinoside and hydroxyurea, or, for example, the antimetabolite described in European Patent Application No. 239362, such as Ν- (5- [Ν (3,4-dihydro-2-methyl-4-oxoquinazoline- 6-ethyl) -Y-methylamino] -2-tenoyl) -E-glutamic acid; growth factor inhibitors; cell cycle inhibitors; intercalating antibiotics, for example adriamycin and bleomycin; enzymes, for example interferon; and anti-hormones, for example anti-estrogens, such as No. 1tabech ™ (tamoxifen) or, for example, antiandrogens, such as Sacobech ™ (4'-cyano-3- (4fluorophenylsulfonyl) -2-hydroxy-2-methyl-3 '- (trifluoromethyl) propionanilide). Such joint treatment can be carried out by simultaneous, sequential or separate dosing of the individual components of the treatment.

The compounds and compositions described in this invention (and, if appropriate,

- 38 020624 chemotherapeutic agent and / or radiation), can be administered in parallel (for example, simultaneously, essentially simultaneously or within the same treatment protocol) or sequentially, depending on the nature of the disease, the condition of the patient and the particular choice of chemotherapeutic agent and / or radiation, which will be administered in combination (i.e., within the same treatment protocol) with the compound / composition.

In combined applications and uses, the compound / composition and chemotherapeutic agent and / or radiation need not be administered simultaneously or essentially simultaneously, and the initial order of administration of the compound / composition and chemotherapeutic agent and / or radiation may not be important. Thus, the compounds / compositions of the invention may be administered first, followed by the administration of a chemotherapeutic agent and / or radiation; or a chemotherapeutic agent and / or radiation may be administered first followed by the administration of the compounds / compositions of the invention. Such an alternative administration may be repeated during a single treatment protocol. Determining the order of administration, and the number of repetitions of the introduction of each therapeutic agent during the treatment protocol, is the responsibility of the attending physician after evaluating the disease being treated and the patient's condition. For example, a chemotherapeutic agent and / or radiation may be administered first, especially if it is a cytotoxic agent, and then treatment is continued by administering the compounds / compositions of the invention, followed by, if deemed favorable, the administration of a chemotherapeutic agent and / or radiation, and such actions are repeated until the completion of the treatment protocol. Thus, according to experience and knowledge, the attending physician can modify each protocol for the administration of the compound / composition for treatment in accordance with the needs of the patient, for the needs of treatment. The attending physician, who must evaluate whether treatment is effective in administering these doses, will take into account the patient’s general well-being, as well as more accurate symptoms, such as relief of symptoms associated with the disease, inhibition of tumor growth, real reduction in tumor, or inhibition of metastases. Tumor size can be measured using standard methods, such as radiological examinations, such as computed axial tomography or MRI scans, and subsequent measurements can be used to determine if tumor growth is slowing or even the tumor is developing back. Relieving symptoms associated with the disease, such as pain, and improving the general condition can also help evaluate the effectiveness of the treatment.

Specific, non-limiting examples of possible combination therapies are the use of the compounds of the invention with agents corresponding to the following pharmacotherapeutic classification below. This list should not be construed as exhaustive, but should be considered as illustrative examples inherent in the relevant therapeutic field at present. In addition, combined treatment regimens may include various routes of administration and will include oral, intravenous, intraocular, subcutaneous, cutaneous, and topical inhalation.

For the treatment of oncological diseases of proliferative disorders and malignant neoplasms, the compounds of the present invention can be administered with an agent selected from the group consisting of aromatase inhibitors, antiestrogen, antiandrogen, corticosteroids, gonadorelin agonists, topoisomerase inhibitors 1 and 2, active agents acting on microtubules, alkylating agents , nitrosoureas, anti-blastoma antimetabolites, compounds containing platinum, targeting lipid or protein kinases, ΙΜίΌ, n targeting agents for protein or lipid phosphatases, antiangiogenic agents, Ak1 inhibitors, ΙΟΡ-Ι inhibitors, POP3 modulators, tTOK inhibitors, mimetics, NEAC inhibitors, drugs that induce cell differentiation, bradykinin 1 receptor antagonists, angiotensin antagonists, cyclooxygenase inhibitors, heparanase inhibitors, lymphokine inhibitors, cytokine inhibitors, 1KK inhibitors, P38MAPK inhibitors, AKKU-797, H8P90 inhibitors, multikinase inhibitors, bisphosphanates, rapamycin derivatives, antiapoptotic inhibitors of the metabolic pathway, apoptotic metabolic pathway agonists, PPAC agonists, AS agonists, Kaz isoform inhibitors, telomerase inhibitors, protease inhibitors, metalloproteinase inhibitors, aminopeptidase inhibitors, §H1P activators - ACX-MM00, Nitah-Sb20, ofum, ofum Biphtheria toxin.

For the treatment of oncological diseases of proliferative disorders and malignant neoplasms, the compounds of the present invention can be administered with an agent selected from the group consisting of dacarbazine (ET1C), actinomycins C ₂ , C ₃ , Ό and P _b cyclophosphamide, melphalan, estramustine, maytansinol, rifamycin, strepticin, strepticin , doxorubicin, daunorubicin, epirubicin, idarubicin, detorubicin, carminomycin, idarubicin, epirubicin, ezlrubitsin, mitoxantrone, bleomycins A, A ₂ and B, camptothecin, irinotekan.KTM., topotekan.KTM., 9 aminokamptote in, 10,11 methylenedioxycamptothecin, 9-nitro-camptothecin, botresomib, temozolomide, TA8103, ΝΓΙ0052, combretastatin, combretastatin A-2, combretastatin A-4, calicheamicins, neocarcinostatins, epothilones A B, Cetanus and roman cactus. KTM., SE40 antibodies, asparaginase, interleukins, interferons. leuprolide, and pegaspargase, 5-fluorouracil, fluorodeoxyuridine, ptorafur, 5'-deoxyfluoruridine, ИРТ, М1ТС, 8-1 capecitabine, diethylstilbestrol, tamoxifen, toremifene, tolmudex, flutamidexonefludimeridone, fluoximetrefdeloim, flufestestonefludimethylfludimeside, fluoximetrodimethylfludimethidefludimeside, fluoximetrodimethylfludimeridonefludimeside, fluoxide leuprorelin acetate, estramustine, droloxifene, medroxyprogesterone, megesterol acetate, aminoglutetimide, testolactone, testosterone, diethylstilbestrol, hydroxyprogesterone, mitomycins A, B and C, porphyromycin, cisplatin oxplatin, carboplatin, carboplatin, carboplatin atine, thalidomide, lenalidomide, C1-973, telomestatin, CH1K258, Cab 001, 8ANA, tubacin, 17-AAS, sorafenib, 1M-216, podophyllotixin, epipodophyllotixin, etoposide, teniposide, Tarceva.KTM. and ress.KTM., imatinib.KTM., miltefosine.KTM., perifosin.KTM., aminopterin, methotrexate, metopterin, dichloromethotrexate, 6-mercaptopurine, thioguanine, azattuoprine, allopurinol, cladribine, fludostinodinine, podudoradinidine, podudaradinin, 2, podoradiatin, podudarabidin, 2-doradinadin, 2 , cytosine arabinoside, cytarabine, azacytidine, 5-azacytosine, gencitabine, 5-azacytosine-arabinoside, vincristine, vinblastine, vinorelbine, leurosin, leurosidine and vindesine, paclitaxel, taxotere and docetaxel.

For the treatment of inflammatory diseases or pain, the compounds of the present invention can be administered with an agent selected from the group consisting of corticosteroids, non-steroidal anti-inflammatory drugs, muscle relaxants and their combinations with other drugs, painkillers and their combinations with other drugs, expectorant drugs and their combinations with others agents, antidepressants, anticonvulsants, and combinations thereof; antihypertensive agents, opioids, local cannabinoids, capsaicin, betamethasone dipropionate (expanded and unexpanded), betamethasone valerate, clobetasol propionate, prednisone, methyl prednisolone, diflorasone diacetate, halobetasol propionate, amcinonide, dexamethasone, desoximetasone, fluocinolone atsetoninid, fluocinonide, galotsinonid, klokortalon pivalate , deoxymethasone, flurandrenalide, salicylates, ibuprofen, ketoprofen, etodolac, diclofenac, sodium meclofenamate, naproxen, piroxicam, celecoxib, cyclobenzaprine, baclofen, cyclobenz aprin / lidocaine, baclofen / cyclobenzaprine, cyclobenzaprine / lidocaine / ketoprofen, lidocaine, lidocaine / deoxy-O-glucose, prilocaine, EMBA cream (eutectic mixture of local anesthetics (lidocaine 2.5% and prilocaine 2.5%), guaifenesin , guaifenesin / ketoprofen / cyclobenzaprine, amitriptyline, doxepin, desipramine, imipramine, amoxapine, clomipramine, nortriptyline, protriptyline, duloxetine, mirtazepine, nizoxetine, maprotiline, reboxetine, fluazetamine lamazinefinfinamin carbazine lamazine lamazinefinfinbamine topoxetaminbazinfinbamenfinamin carbazine laminate mesipine, zonisamide, mexiletine, gabapentin / clonidine, gabapentin / carbamazepine, carbamazepine / cyclobenzaprine, antihypertensive drugs, including clonidine, codeine, loperamide, tramadol, morphine, fentanyl, oxycodone, hydrocodonefol, efolofentol, camphoronol evolvefan, camphoronol evolvefan, camphoronol evolvefan, camphoronol, morpholum, hydrochlorothiazole, morpholum, hydrochlorothiazone, hydrochlorothiazol oil, turpentine oil; CB1 / CB2 ligands, acetaminophen, infliximab, nitric oxide synthase inhibitors, in particular inhibitors of inducible nitric oxide synthase, ΡΌΕ4 inhibitors - a mechanism similar to ibudilast (AU-411), SIS-801, ΊΝΚ inhibitors - SS-401, combination T№ / REE4 inhibitors - SES-998, 1L1 antagonists, for example Apak1ita - Ksheteb AMS 108, (tAb), which are aimed at 1L-1, 8H1P activators - ArX-MM00, C5 antagonists, C5a inhibitors, pexelizumab, pyrimidine synthesis inhibitors, cytokine inhibitors , ΙΚΚ inhibitors, P38MAPK inhibitors, AKKU-797, H8P90 inhibitors, ing multikinase inhibitors, bisphosphonates, PPAC agonists, Cox 1 and Cx 2 inhibitors, anti-CE4 therapy, B-cell inhibitors, COX / Lox dual inhibitors, immunosuppressive drugs, δ inhibitors, NSAIDs, 8PA2 inhibitors, colchicine, allipurinol, oxipurinol, Yubaita - Aigaioyi, febucostat, puricase, RES-uricase preparations, benzbromaron, beta-2 long-acting agonists (PABA), salmeterol (8eteuye E18ki8) and formoterol (RogabP), leukotriene modifiers, including montelukast (8difluaeaa) inhaled cromolyn (1i1a1) or undercromium l (TIAbe), theophylline. short-acting beta-2 agonists, ipratropium (Abouey), immunotherapy- (allergen-specific immunotherapy), anti-1de monoclonal antibodies - Ho1ab, generally accepted basic antirheumatic drugs that modify the course of the disease (EMAKE), including hydroxychloroquine (P1adiy1), gold aurano compounds (Rtbaita), sulfasalazine (A / ybbsche), minocycline (Eurascash, Mosshosh) and methotrexate (Kbeitabeh), leflunomide (Ataua), azathioprine (1tigai), cyclosporine (No. 1, 8aib1ttie) and Cyclophosphamide, (Cytophosphamide, 80) costimulir antagonists factor, Nitah-SE20 (ofatumumab); CE20 antagonists, MEK inhibitors, ΝΡ kappa B inhibitors, antibodies to B cells, denosumab, tAb, which specifically targets the ligand of the kappa B nuclear factor receptor activator ligand (KAMKE), antibodies that inactivate 1b17, 1b-17 receptor antagonists / inhibitors, CTLA inhibitors, CE20 inhibitors, soluble UESPK-1 receptors, antibodies to the UESPK1 receptor, antibodies to UESP, integrin receptor antagonist, selectin inhibitors, P-selectin and E-selectin inhibitors, phospholipase A2 inhibitors, lipoxygenase inhibitors, KA antagonists and antibodies , en osteoprotegerin agonists, lymphotoxin inhibitors, B-lymphocyte stimulator, MCP-1 inhibitors, M1P inhibitors, inhibitors for CE2, SE3, SE4, SE25, SE40 and SE40 ligand SI 152 (CTA4), macrilide immunosuppressants, inhibitors, selective inhibitors, nucleotide inhibitors, inhibitors CXC receptor and CXC ligand inhibitors, chemokine antagonists, leukocyte chemotaxis inhibitors, adhesion molecule blockers, leukocyte antigen-1 antagonists

- 40 020624 selectin (LRA-1, CE11a), antagonists of very late antigen-4 (νΓΛ-4), matrix metalloprotease inhibitors, elastase inhibitors, cathepsin inhibitors.

For the treatment of ophthalmic disorders and eye diseases, the compounds of the present invention can be administered with an agent selected from the group consisting of beta-blockers, carbonic anhydrase inhibitors, .alpha. And .beta. Adrenergic antagonists, including alpha-adrenergic antagonists, .alpha. 2 agonists, miotic agents, prostaglandin analogues, corticosteroids and immunosuppressive drugs.

For the treatment of ophthalmic disorders and eye diseases, the compounds of the present invention can be administered with an agent selected from the group consisting of timolol, betaxolol, levobetaxolol, cartolol, levobunolol, propranolol, brinzolamide, dorzolamide, nipradilil, iopidine, brimonidine, pilocarpinoprine, latin epine, pilocarpinopin, epinoprostin, trapoprinopin, latin , bimatoprost, unoprostone, dexamethasone, prednisone, methylprednisolone, azathioprine, cyclosporine and immunoglobulins.

For the treatment of autoimmune disorders, the compounds of the present invention can be administered with an agent selected from the group consisting of corticosteroids, immunosuppressive agents, prostaglandin analogs and antimetabolites.

For the treatment of autoimmune disorders, the compounds of the present invention can be administered with an agent selected from the group consisting of dexamethasis, prednisone, methylprednisolone, azathioprine, cyclosporin, immunoglobulins, latanoprost, travoprost, bimatoprost, unoproston, infliximab, rutuximab, methotrexantel mesterol, non-mediastinospide medications, not their combinations with other agents, painkillers and their combinations with other agents, expectorants and their combinations with other agents, anti depressants, anticonvulsants, and combinations thereof; antihypertensive drugs, opioids, local cannabinoids, and other drugs, such as capsaicin, betamethasone dipropionate (expanded and unexpanded), betamethasone valerate, clobetasol propionate, prednisone, methyl prednisolone, diflorazone diacetate, halobetazole propionate, aminone diacetone, acetone diacidone, acetone dimethasone, acetone dimethasone, acetone dimethasone, acetone dimethasone, acetone, acetone fluocinonide, haloconid, clocortalon pivalate, deoxymethasone, flurandrenalide, salicylates, ibuprofen, ketoprofen, etodolac, diclofenac, sodium meclofenamate, naproxen, piroxicam, celecoxib, cyclobe nzaprine, baclofen, cyclobenzaprine / lidocaine, baclofen / cyclobenzaprine, cyclobenzaprine / lidocaine / ketoprofen, lidocaine, lidocaine / deoxy-O-glucose, prilocaine, EMBA cream (eutectic mixture of local anesthetics (lidocaine 2.5% iloca priloca) %)), guaifenesin, guaifenesin / ketoprofen / cyclobenzaprine, amitriptyline, doxepin, desipramine, imipramine, amoxapine, clomipramine, nortriptyline, protriptyline, duloxetine, mirtazepine, reboxinetaminfinetamilafineplaminfinfetilaminfinfetilaminfinfetilaminfin teflin tufetamilinfetamilinfinetilfinfilaminfin teflinfelotinfin t agabin, oxcarbazepine, karbamezipin, zonisamide, mexiletine, gabapentin / clonidine, gabapentin / carbamazepine, carbamazepine / cyclobenzaprine, antihypertensives including clonidine, codeine, loperamide, tramadol, morphine, fentanyl, oxycodone, hydrocodone, levorphanol, butorphanol, menthol, gaulterievoe oil camphor, eucalyptus oil, turpentine oil; CB1 / CB2 ligands, acetaminophen, infliximab; nitric oxide synthase inhibitors, in particular inhibitors of inducible nitric oxide synthase; and other drugs, such as capsaicin. ΡΌΕ4 inhibitors - a mechanism similar to ibudilast (A ^ 411), SES-801, ΊΝΚ inhibitors - SS-401, the combination of ΤΝΡ / ΡΌΕ4 inhibitors - SES-998, 1b1 antagonists for example Apak1nga - Ktegek AMC 108, (tAb), which target 1b -1, δΗΙΡ activators - AOX-100, C5 antagonists, C5a inhibitors, pexelizumab, pyrimidine synthesis inhibitors, lymphokine inhibitors, cytokine inhibitors, Р inhibitors, P38MA6K inhibitors, ACC ¥ -797, ΗδΡ90 inhibitors, multifunctional agonists, bisulfate kinase inhibitors, cx2 inhibitors, anti-CE4 therapy, b-cell inhibitors, COX / Lox dual iterators, immunosuppressive agents, ίΝΘδ inhibitors, NSAIDs, 8Ρ ^ A2 inhibitors, colchicine, allopurinol, oxypurinol, Co1b, Kzbaiga - Aigapoyp, febucostat, puricase, ΡΕC-uricase preparations, benzbromaron, beta-2 agonists, long-acting salamer (ЕА) (ЕА) EGUESH EKIC and formoterol (Rogabb), leukotriene modifiers, including montelukast (Ypdikig) and zafirlukast (Asso1a1e), inhaled cromoline (1p1a1) or nedocromil (Tbabe), theophylline. short-acting beta-2 agonists, ipratropium (Ayoui), immunotherapy (allergen-specific immunotherapy), anti-1§E monoclonal antibodies - Ho1ab, generally accepted basic antirheumatic drugs that modify the course of the disease (EMAKE), including hydroxychloroquine (Ρ ^^ ώ1) , gold compounds auranofin (Shbaiga), sulfasalazine (A / iShbsche), minocycline (Euphrasch, Mshossh) and methotrexate (Kieitayeh), leflunomide (Agaua), azathioprine (1tigap), cyclosporine (Nooga1, δapb ^ ttyphamide) and ), antibiotics, CE80 antagonists, antagonists costimulatory of factor nitah-SE20 (ofatumumab); CE20 antagonists, MEK inhibitors, ΝΡ kappa B inhibitors, antibodies to B cells, denosumab, tAb, which specifically targets the ligand of the Kappa B nuclear factor receptor activator ligand (KAIK), antibodies that inactivate 1b17, antagonists / inhibitors of the 1b-17 receptor, CTbA inhibitors, CE20 inhibitors, soluble receptors of VΕΟPΚ-1, antibodies to the VΕΟPΚ-1 receptor, antibodies to νΈΟΡ, integrin receptor antagonist, selectin inhibitors, P-selectin and E-selectin inhibitors, phospholipase A2 inhibitors, lipoxygenase inhibitors, antagonists and KAIK, and tagonisty osteoprotegerin inhibitors fotoksina 41 020624 lymphoma, B-lymphocyte stimulator, MCP-1 inhibitors, M1R inhibitors, for: SE2. SEZ. SE4, SE25, SE40, and SE40 CO 152 ligand (CTLA4), macrilide immunosuppressants, selective nucleotide metabolism inhibitors, chemotaxis inhibitors, CXC receptor and CXC ligand inhibitors, chemokine antagonists, leukocyte leukocyte antagonists, antagonists of leukocyte leukocyte antagonists, LRA-1, CE11a), antagonists of very late antigen-4 (UL-4), matrix metalloprotease inhibitors, elastase inhibitors, cathepsin inhibitors.

For the treatment of metabolic disorders, the compounds of the present invention can be administered with an agent selected from the group consisting of insulin, insulin derivatives and mimetics, insulin secretion stimulants, insulin sensitizers, biguanides, alpha aglucosidase inhibitors, insulinotropic sulfonylurea receptor ligands, and protein R -ase inhibitors 1B), O8K3 inhibitors (kinase-3 of glycogen synthase), ОРР-1 (glucagon-like peptide-1), ОРР-1 analogues, ΌΡΡίν inhibitors (dipeptidyl peptidase IV), K inhibitors HC of ligands of the sodium-dependent glucose cotransporter, glycogen phosphorylase A inhibitors, AOE disruptor, PPAC modulators, LXK and PXK modulators, non-glitazone type agonist PRAK8, selective glucocorticoid antagonists, metformin, glipizide, gliburide, amnidyl, amnidyl, amdylidene 8В-517955, 8В4195052, 8В-216763, NN-57-05441, NN-57-05445, 0ν-0791, ΑΟΝ.5pyr.194.5ir.204, T-1095, ΒΑΥ K3401, exendin-4 acarbose, ORP728, LAR237 , vildagliptin, MK-0431, saxagliptin, O8K23A, pioglitazone, rosiglitazone, (K) -1- {4- [5-methyl-2- (4-trifluoromethylphenyl) oxazol4-ylmethoxy] benzenesulfonyl} -2, The 3-dihydro-1H-indole-2-carboxylic acid described in νθ 03/043985, as compound 19 in example 4, and 01-262570.

Diseases

The present invention describes methods of treating a disease or disorder in an individual suffering from said disease or disorder, which comprises administering to said individual an effective amount of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) - 6-methoxyphenyl) -1 (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (Form A). The present invention covers the use of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (Form A) for preparing a medicament for treating a disease or disorder.

In some embodiments, the invention relates to the prevention or treatment of any disease or disorder in which MEK kinase is involved, including, but not limited to, oncological, hematological, inflammatory, ophthalmic, neurological, immunological, cardiovascular and dermatological diseases, and also diseases caused by excessive or unregulated production of pro-inflammatory cytokines, including, for example, excessive or unregulated production of TOT ¹ , 1b-1, 1b-6 and 1b -8 in humans or other mammals. The invention encompasses such use and use of compounds for the preparation of a medicament for the treatment of such diseases or disorders mediated by cytokines. The invention further encompasses the administration to humans of an effective amount of an MEK inhibitor for the treatment of such a disease or disorder.

Diseases or disorders in which MEK kinase is involved, either directly or through proinflammatory cytokines, including kittoks TOT ¹ , 1L-1, 1L-6 and 1L-8, include, without limitation, dry eyes, glaucoma, autoimmune diseases, inflammatory diseases, destructive bone disorders, proliferative disorders, neurodegenerative disorders, viral diseases, allergies, infectious diseases, myocardial infarction, angiogenic disorders, reperfusion / ischemia upon impact, vascular hyperplasia, organ hypoxia, hypertension raphy infarction, thrombin-induced platelet aggregation, and conditions associated with prostaglandin endoperoksidazoysintazoy-2 (COX-2).

In certain aspects of the invention, the disease is a hyperproliferative state of a human or animal, including, but not limited to, malignant neoplasm, hyperplasia, restenosis, inflammation, immune disorders, myocardial hypertrophy, atherosclerosis, pain, migraine, conditions or disorders associated with angiogenesis, proliferation induced after diseases, including, but not limited to, vascular plastic surgery, or other conditions.

In further embodiments, said hyperproliferative state is selected from the group consisting of hematologic and non-hematologic malignancies. In further embodiments, said hematologic malignancy is selected from the group consisting of multiple myeloma, leukemia, and lymphoma. In further embodiments, said leukemia is selected from the group consisting of acute and chronic leukemia. In further embodiments, said leukemia is selected from the group consisting of acute lymphocytic leukemia (ALL) and acute non-lymphocytic leukemia (ONLL). In further embodiments, said chronic leukemia is selected from the group consisting of chronic lymphocytic leukemia (CLL) and chronic myelogenous leukemia (CML). In further embodiments, the indicated lymphoma is selected from the group consisting of Hodgkin's lymphoma and non-Hodgkin's lymphoma. In further embodiments, said hematologic malignancy is multiple myeloma. In other embodiments, said hematologic malignant neoplasm is a low-grade, a low-grade, or a high-grade. In other embodiments, said non-hematologic malignant neoplasm is selected from the group consisting of brain cancer, head and neck cancer, lung cancer, breast cancer, genital cancer, digestive cancer, pancreatic cancer, and urinary cancer. In further embodiments, said cancer of the digestive system is cancer of the upper gastrointestinal tract or cancer of the colon and rectum. In further embodiments, said urinary system cancer is bladder cancer or renal cell carcinoma. In further embodiments, said genital cancer is prostate cancer.

Additional types of malignant neoplasms that can be treated with the compounds and methods described in this invention include malignant neoplasms of the oral cavity and pharynx, malignant neoplasms of the respiratory system, malignant neoplasms of the bones and joints, malignant neoplasms of soft tissues, skin cancer, cancer of the reproductive system, malignant neoplasms of the eyes and ocular orbit, malignant neoplasms of the nervous system, malignant neoplasms l imphatic system, and malignant neoplasms of the endocrine system. In certain embodiments, these malignancies may be selected from the group consisting of cancer of the tongue, mouth, pharynx, or other parts of the oral cavity; esophageal cancer, stomach cancer, or cancer of the small intestine; cancer of the colon or rectum, anus, or anorectal cancer; cancer of the liver, intrahepatic bile ducts, gall bladder, pancreas, or other bile or digestive organs; cancer of the larynx, bronchi, and other malignant neoplasms of the respiratory system; heart cancer, melanoma, basal cell carcinoma, squamous cell carcinoma, other non-epithelial skin cancer; cancer of the uterus or cervix; cancer of the uterus; cancer of the ovaries, external female genital organs, vagina or other female genital organs; cancer of the prostate, testicles, penis, or other male genital organs; bladder cancer; renal cancer; cancer of the kidney, pelvis or urethra or other malignant neoplasm of the genitourinary organs; thyroid cancer or other endocrine malignancy; chronic lymphocytic leukemia and cutaneous T-cell lymphoma, both granulocytic and monocytic.

Other types of malignant neoplasms that can be treated with the compounds and methods described in this invention are adenocarcinoma, malignant hemangioma, astrocytoma, auditory nerve neuroma, anaplastic astrocytoma, basal cell carcinoma, blastoglioma, chondrosarcoma, choriocarcinoma, kochiofarcinoma, kochiofarcinoma, kochiofarcinoma, kochiofarcinoma, kohriofarcinoma, kohriokarcinoma, kochiofarcinoma, kochiofarcinoma, kochiofarcinoma, kochiofarcinoma, kochiofarcinoma, kochiofarcinoma, kochiokarcinoma, kokhina melanoma, cystadenocarcinoma, endotheliosarcoma, embryonic cancer, ependymoma, Ewing's sarcoma, epithelial carcinoma, fibrosarcoma, gastric cancer, urinary tract cancer, multif rhyme glioma, hemangioblastoma, hepatocellular carcinoma, hepatoma, Kaposi’s sarcoma, large cell carcinoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, medullary thyroid cancer, medullary fibrosomeglioma myelomegrobomelomerosomerosomerosis ovarian cancer, papillary cancer, papillary adecarcinoma, parathyroid tumors, pheochromocytoma, pinealoma, plasmacytoma, retinoblastoma, rhabdomyosarcoma, fat cancer gland, seminoma, malignant neoplasms of the skin, melanoma, small cell lung cancer, squamous cell carcinoma, sweat cancer, synovioma, thyroid cancer, uveal melanoma and Wilms tumor.

The present invention also provides methods for treating a hyperproliferative disorder in a mammal, which comprises administering to the said mammal a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, solvate, polymorph, ester, amide, tautomer, prodrug, hydrate or derivative thereof, in combination with an antitumor means. In some embodiments, the antitumor agent is selected from the group consisting of mitosis inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, anti-androgens, anti-androgens - Α0Χ-ΜΝ100. Nntah-SE20 (ofatumumab), SE20 antagonists, fusions of the L2-diphtheria toxin.

A disease that can be treated with the compounds, compositions and methods described in this invention may be a hematologic disorder. In certain embodiments, said hematologic disorder is selected from the group consisting of sickle cell anemia, myelodysplastic disorders (ΜΌδ), and myeloproliferative disorders. In further embodiments, said myeloproliferative disorder is selected from the group consisting of true polycythemia, myelofibrosis, and idiopathic thrombocythemia.

- 43,020,624

The compounds, compositions and methods described in this invention can be useful as anti-inflammatory drugs with the additional advantage of substantially less side effects. The compounds, compositions and methods described in this invention are suitable for the treatment of arthritis, including, but not limited to, rheumatoid arthritis, spondyloarthropathy, ankylosing spondylitis, gout, gouty arthritis, osteoarthritis, systemic lupus erythematosus, Still's disease, acute rheumatoid arthritis enteropathic arthritis, neuropathic arthritis, psoriatic arthritis and purulent arthritis. The compounds, compositions and methods described in this invention are also suitable for the treatment of osteoporosis and other related bone lesions. Such compounds, compositions and methods described in this invention are also suitable for the treatment of gastrointestinal disorders such as esophagitis, diarrhea, inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel syndrome and ulcerative colitis. The compounds, compositions and methods described herein may also be useful in the treatment of pneumonia, such as those associated with viral infections and cystic fibrosis. Additionally, the compounds, compositions and methods described in this invention are also suitable for the treatment of patients with transplanted organs, either alone or in combination with conventional immunomodulators. Additionally, the compounds, compositions and methods described in this invention are useful for the treatment of pruritus and vitiligo. In particular, the compounds, compositions and methods as described herein are useful for treating a specific inflammatory disease of rheumatoid arthritis.

Further inflammatory diseases that can be prevented or treated include, but are not limited to, asthma, allergies, respiratory distress syndrome, or acute or chronic pancreatitis. In addition, diseases of the respiratory system that can be prevented or treated include, but are not limited to, chronic obstructive pulmonary disease, and pulmonary fibrosis. Additionally, MEK kinase inhibitors, as described in this invention, are also associated with the production of prostaglandin endoperoxidase synthase-2 (COX-2). Pro-inflammatory mediators of the cyclooxygenase metabolic chain, arachidonic acid derivatives, such as prostaglandins, are produced using an inducible COX-2 enzyme. Regulation of COX-2 will regulate these pro-inflammatory mediators that affect various cells and are important and critical mediators of inflammation of various disease states and conditions. In particular, these inflammatory mediators are involved in pain, such as sensitization of pain receptors, and edema. Therefore, additional MEK kinase-mediated conditions that can be prevented or treated include edema, analgesia, fever, and pain such as neuromuscular pain, headache, toothache, arthritis pain, and pain caused by malignancy.

Additionally, a disease that can be treated with the compounds, compositions and methods described in this invention may be an ophthalmic disorder. Ophthalmic diseases and other diseases in which angiogenesis can be treated or prevented include, without limitation, dry eyes (including Sjogren's syndrome), macular degeneration, angle-closure and open-angle glaucoma, retinal ganglion degeneration, eye ischemia, retinitis, retinopathy, uveitis, ocular photophobia, and inflammation and pain associated with acute damage to the eye tissue. The compounds, compositions and methods described in this invention are useful for the treatment of glaucoma retinopathy and / or diabetic retinopathy. The compounds, compositions and methods described in this invention are also useful for the treatment of postoperative inflammation or pain in eye surgery, such as cataract surgery and refractive surgery. In further embodiments, said ophthalmic disorder is selected from the group consisting of dry eyes, closed-angle glaucoma, and open-angle glaucoma.

In addition, a disease that can be treated with the compounds, compositions and methods described in this invention may be an autoimmune disease. Autoimmune diseases that can be prevented or treated include, but are not limited to rheumatoid arthritis, inflammatory bowel disease, inflammation pain, ulcerative colitis, Crohn's disease, periodontal disease, temporomandibular joint disease, multiple sclerosis, diabetes, glomerulonephritis , systemic lupus erythematosus, scleroderma, chronic thyroiditis, diffuse toxic goiter, hemolytic anemia, autoimmune gastritis, autoimmune neutropenia, thrombocytopenia, chro active hepatitis, asthenic bulbar palsy, atopic dermatitis, graft versus host disease, and psoriasis. Inflammatory diseases that can be prevented or treated include, but are not limited to, asthma, allergies, respiratory distress syndrome, or acute or chronic pancreatitis. In particular, the compounds, compositions and methods, as described in this invention, are useful for the treatment of specific autoimmune diseases - rheumatoid arthritis and multiple sclerosis.

Additionally, a disease that can be treated with the compounds, compositions and methods described in this invention may be a dermatological disorder. In certain embodiments, said dermatological disorder is selected from the group consisting of,

- 44 020624 but not limited to melanoma, basal cell carcinoma, squamous cell carcinoma and other non-epithelial malignant neoplasms of the skin, such as psoriasis and persistent itching, and other diseases associated with the skin and skin structures that can be treated or prevented with MEK kinase inhibitors according to the present invention.

Metabolic diseases that can be treated or prevented include, without limitation, metabolic syndrome, insulin resistance, and type 1 and type 2 diabetes. Additionally, the compositions described in this invention may be useful for treating insulin resistance and other metabolic disorders, such as atherosclerosis, which are usually associated with pathologically increased transmission of inflammatory signals.

The compounds, compositions and methods described in this invention are also suitable for the treatment of tissue damage in diseases such as vascular diseases, migraine headaches, periarteritis nodosa, thyroiditis, aplastic anemia, Hodgkin's disease, sclerodoma, rheumatic attack, type I diabetes, neuromuscular junction disease, including asthenic bulbar palsy, white matter disease, including multiple sclerosis, sarcoidosis, nephritis, nephrotic syndrome, Behcet syndrome, polymyositis, gingivitis, periodontitis, hypersensitivity, swelling that develops after trauma, ischemia, including myocardial ischemia, cardiovascular ischemia, and ischemia secondary to cardiac arrest, etc. Compounds, compositions and methods described in this invention may also be useful for treatment allergic rhinitis, respiratory distress syndrome, endotoxin shock syndrome and atherosclerosis.

Additionally, the disease that can be treated using the compounds, compositions and methods described in this invention may be a cardiovascular condition. In certain embodiments, said cardiovascular condition is selected from the group consisting of atherosclerosis, myocardial hypertrophy, idiopathic cardiomyopathies, heart failure, conditions or disorders associated with angiogenesis, and proliferation induced after diseases, including, but not limited to, restenosis after surgical intervention and plastic surgery on the vessels.

Additionally, a disease that can be treated with the compounds, compositions and methods described in this invention may be a neurological disorder. In certain embodiments, said neurological disorder is selected from the group consisting of Parkinson's disease, Alzheimer's disease, Alzheimer's dementia, and damage to the central nervous system due to stroke, ischemia, and trauma. In other embodiments, said neurological disorder is selected from the group consisting of epilepsy, neuropathic pain, depression, and bipolar disorder.

Additionally, a disease that can be treated using the compounds, compositions and methods described in this invention can be a malignant neoplasm, such as acute myeloid leukemia, cancer of the thymus, brain, lung, squamous cells, skin, eyes, retinoblastoma, intraocular melanoma, oral cavity and oropharynx, bladder, stomach, stomach, pancreas, bladder, breast, cervix, head, neck, kidneys, kidney, liver, ovaries, prostate, colorectal cancer, food and, testis, genital system, thyroid, CNS, PNS, AIDS-related (e.g. Lymphoma and Kaposi's sarcoma) or virus-induced. In some embodiments, the compounds and compositions are intended for the treatment of a non-malignant hyperproliferative disorder, such as benign skin hyperplasia (e.g., psoriasis), restenosis, or the prostate (e.g., benign prostatic hypertrophy (BPH)).

Additionally, a disease that can be treated with the compounds, compositions and methods described in this invention can be pancreatitis, kidney disease (including proliferative glomerulonephritis and kidney disease induced by diabetes), pain, a disease associated with vasculogenesis or angiogenesis, tumor angiogenesis chronic inflammatory disease such as rheumatoid arthritis, inflammatory bowel disease, atherosclerosis, skin diseases such as psoriasis, eczema, and scleroderma, diabetes, etc. diabetic retinopathy, premature retinopathy, senile macular degeneration, hemangioma, tendonitis, bursitis, sciatica, glioma, melanoma, Kaposi’s sarcoma and cancer of the ovaries, breast, lungs, pancreas, prostate, colon and epidermoid cancer in a mammal.

Additionally, a disease that can be treated with the compounds, compositions and methods described in this invention may be the prevention of implantation of blastocytes in a mammal.

Patients who can be treated with the compounds described in this invention, or their pharmaceutically acceptable salts, solvates, polymorphs, esters, amides, tautomers, prodrugs, hydrates or derivatives in accordance with the methods in accordance with the present invention include, for example, patients diagnosed with psoriasis; restenosis; atherosclerosis; BPH; breast cancer, such as carcinoma from ductal epithelium into duct tissue in the mammary gland, medullary carcinomas, colloid carcinomas, tubular carcinomas, and inflammatory cancer

- 45 020624 mammary gland; ovarian cancer, including ovarian epithelial tumors such as adenocarcinoma in the ovary and adenocarcinoma that migrated from the ovary to the abdominal cavity; uterine cancer; cervical cancer, such as adenocarcinoma in the epithelium of the cervix, including squamous cell carcinoma and adenocarcinomas; prostate cancer, such as prostate cancer, selected from the following: adenocarcinoma or adenocarcinoma that migrated to the bone; pancreatic cancer, such as epithelioid carcinoma in the tissue of the pancreatic duct and adenocarcinoma in the pancreatic duct; bladder cancer, such as transitional cell carcinoma in the bladder, urothelial carcinomas (transitional cell carcinoma), tumors in urothelial cells that are located in the bladder, squamous cell carcinoma, adenocarcinomas, and small cell carcinoma; leukemia, such as acute myelogenous leukemia (AML), acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, myelodysplasia and myeloproliferative disorders; bone cancer lung cancer, such as non-small cell lung cancer (CBC), which is divided into squamous cell cancer, adenocarcinomas, and undifferentiated large cell cancer, and small cell lung cancer; skin cancer, such as basal cell carcinoma, melanoma, squamous cell carcinoma, and actinic keratosis, which is a skin condition that sometimes develops into squamous cell carcinoma; retinoblastoma; cutaneous or intraocular (ophthalmic) melanoma; primary liver cancer (cancer that occurs in the liver); kidney cancer thyroid cancer, such as papillary, follicular, medullary and anaplastic; AIDS-related lymphoma, such as diffuse B-large cell lymphoma, immunoblastic B-cell lymphoma, and small cell lymphoma with unseparated nuclei; Kaposi's sarcoma; virus-induced cancer, including hepatitis B virus (IVV), hepatitis C virus (HCV), and hepatic cell cancer; human lymphotropic virus type 1 (NTLU-1) and adult T-cell leukemia / lymphoma; and human papillomavirus (NRU) and cervical cancer; malignant neoplasms of the central nervous system (CNS), such as a primary brain tumor, which includes gliomas (astrocytoma, anaplastic astrocytoma or glioblastoma multiforme), oligodendroglioma, ependymoma, meningioma, lymphoma, neuroma and medulloblastoma; malignant neoplasms of the peripheral nervous system (PNS), such as neuroma of the auditory apparatus and a malignant tumor of the sheath of the peripheral nerves (MR No. T), including neurofibromas and neuromas, malignant fibrotic cytoma, malignant fibrotic histi histitis, malignant meningioma, and malignant mesothelioma and 11; malignant neoplasms of the oral cavity and oropharynx, such as hypopharyngeal cancer, laryngeal cancer, cancer of the nasopharynx, and oropharyngeal cancer; gastric cancer, such as lymphomas, tumors of the stroma of the stomach, and carcinoid tumors; testicular cancer, such embryonic cell tumors (CCTs), which include seminomas and nonseminomas, and gonadal stromal tumors, which include tumors from Leydig cells and tumors from Sertoli cells; malignant neoplasm of the thymus gland, such as thymomas, thymic carcinomas, Hodgkin’s disease, non-Hodgkin’s lymphomas, carcinoids, carcinoid tumors; colorectal cancer and colon cancer.

Sets

The compounds, compositions and methods described in this invention provide kits for treating disorders such as those described in this invention. These kits include the compound, compounds or compositions described in this invention in a container and, optionally, instructions for using the kit according to various methods and approaches as described in this invention. Such kits may also include information, such as links to scientific literature, package leaflet, clinical trial results, and / or summary information from such sources, etc. that indicates or evaluates the activity and / or benefits of the composition, and / or which describe the dosage, administration, side effects, drug interactions, or other information useful to the healthcare provider. Such information can be based on the results of various studies, for example, studies using experimental animals, using the ίη νίνο model, and studies based on clinical trials in humans. Kits, as described herein, may be provided, sold, and / or marketed by medical professionals, including general practitioners, nurses, pharmacists, official guides, etc. Also, in some embodiments, kits may be sold directly to the consumer.

The compounds described in this invention can be used as diagnostic tools and as experimental reagents. For example, the compounds described in this invention, either individually or in combination with other compounds, can be used as tools in differential and / or combinatorial analyzes to identify the expression patterns of genes expressed in cells and tissues. By way of non-limiting example, the expression patterns in cells or tissues treated with one or more compounds are compared with control cells or tissues, untreated compounds, and the resulting patterns are analyzed for differential expression levels of genes to which they relate, for example, to an associated disease , signal transmission pathways, localization in the cell, expression level, size, structure or function of the studied genes. These assays can be performed on stimulated or unstimulated cells and in the presence or absence of other compounds that

- 46,020,624 affect the nature of expression.

In addition to being suitable for treating humans, the compounds and preparations of the present invention are also suitable for veterinary treatment of domestic animals (e.g., dogs, cats), exotic animals and farm animals (e.g., horses), including mammals, rodents, and others.

The examples and preparations presented below further illustrate and exemplify the compounds of the present invention and methods for preparing such compounds. It is implied that the scope of the present invention is in no way limited to the following examples and preparations.

Examples

General typical methods for the synthesis of sulfonamides.

Procedure A. Anhydrous triethylamine (5 equiv.) Was added to a solution of the amine (1 equiv.) In anhydrous dichloromethane (3 ml / mmol). Sulfonyl chloride (1 equiv.) Was added to this solution, and the solution was stirred at room temperature for 16 hours. The solvent was evaporated and the residue was purified by flash column chromatography on silica.

Procedure B. To a stirred solution of an amine (1 equiv.) In anhydrous pyridine (5 ml / mmol) was added sulfonyl chloride (1-5 equiv.). The reaction mixture was stirred at 40 ° C. for 48 hours. The reaction mixture was partitioned between water and EUAc. The organic layer was washed with brine, dried (MO ₈ O ₄ ) and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica.

Procedure C. Substitution of the iodine atom.

A suspension containing 1 eq. aryl iodide, 1.5 eq. boronic acid or boronic ester, 0.25 eq. RyS1 ₂ (yrr £) hDHM and 10 equiv. anhydrous K ₂ CO ₃ powder in an oxygen-free mixture of dioxane and water (3: 1) was heated in a microwave reactor for 60 min at 115 ° C. It was extracted with aq. IN ₄ C1 / THF, and the organic fraction was dried using Να ₂ 80. ₄ . The crude reaction products were purified using flash column chromatography (8ί, EJAc / hexanes or CHC1 ₃ / MeOH). Yields: 20-40%.

Procedure Ό. Synthesis of ^ (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -2 (alkylamino) ethanesulfonamide.

2-Chloroethanesulfonyl chloride (0.1 ml, 1 mmol) was added to a solution of 5,6-difluoro- ¹ - (2-fluoro-4-iodophenyl) benzene-1,2-diamine (0.364 g, 1 mmol) and triethylamine (0, 28 ml, 2 mmol) in CH ₂ Cl ₂ (5 ml) and the reaction mixture was stirred at room temperature for 16 hours. Then it was treated with an excess of amine (10 equiv.) Either in solution or as a pure liquid. The reaction mixture was stirred at room temperature for an additional 6 hours. The reaction mixture was diluted with CH ₂ Cl ₂ (10 ml) and water (10 ml). The organic layer was washed sequentially. HC1 (2x20 ml, 2 N) and saturated Na растворомCO ₃ solution (2x10 ml). After that, CH ₂ Cl ₂ layer was dried (MD8O ₄ ) and evaporated to give a crude product. The contaminated product was purified by preparative HPLC to give pure products in 50-60% yield.

Example 55

^ (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1 sulfonamide

Stage A. 1-Allyl-Y- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-1 sulfonamide

Similarly to General Procedure B, 1-allylcyclopropanesulfonyl chloride was reacted with 5,6difluoro-I1- (2-fluoro-4-iodophenyl) -3-methoxybenzene-1,2-diamine to give the title product.

Ή NMR (ΟϋΟ1 ₃ , 300 MHz): δ 7.417 (s, 1H), 7.309 (5, 1H), 7.25 (t, 1H), 6.89 (t, 1H), 6.52 (t, 1H) ), 6.427 (t, 1H), 6.03 (5, 1H), 5.668 (t, 1H), 5.11 (ΐ, 1H), 3.9 (5, 3H), 2.75 (2H) 1.21 (t, 2H); 0.767 (t, 2H).

- 47,020,624

Stage B. Y- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

1-Allyl-M- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-1-sulfonamide (97 mg, 0.18 mmol) and 4-methylmorpholine Ν-oxide (21 mg, 0.18 mmol) was dissolved in THF (8 ml). Osmium tetroxide was added at room temperature (0.018 mmol, 0.13 ml, 4% in H ₂ O) and the reaction mixture was stirred at room temperature for 16 hours. EJAc was added, the organic phase was washed with water, dried (MDBO ₄ ) and concentrated at reduced pressure. The residue was purified by silica gel chromatography (eluants: EluAc / MeOH) to give the title product (0.80 g, 78%).

Ή NMR (SBSl, 300 MHz): δ 7.38 (nd, 1 = 1.7 and 10.3 Hz, 1H), 7.26 (t, 1H), 7.14 (q, 1H), 6, 87 (q, 1H), 6.53 (nd, 1 = 6.8 and 11.4 Hz, 1H), 6.43 (t, 1H), 4.06 (t, 1H), 3.89 (q , 3H), 3.63 (st, 1 = 3.7 and 11.1 Hz, 1H), 3.49 (st, 1 = 6.4 and 11.1 Hz, 1H), 2.3 (st, 1 = 9.7 and 16.1 Hz, 1H), 1.77 (st, 1 = 1.9 and 16.0 Hz, 1H), 1.37 (t, 1H), 1.25 (t, 1H) ), 1.21 (t, 2H), 0.86 (t, 2H).

t / ζ 571 [M-1] ^- .

Example 56

(B) ^ - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

The pure B isomer was obtained by chiral HPLC resolution of a racemic mixture (Example 55).

Ή NMR (SPS1 ₃ , 300 MHz): δ 7.38 (nd, 1 = 1.7 and 10.3 Hz, 1H), 7.26 (t, 1H), 7.14 (q, 1H), 6 87 (q, 1H), 6.53 (y, 1 = 6.8 and 11.4 Hz, 1H), 6.43 (t, 1H), 4.06 (t, 1H), 3.89 ( q, 3H), 3.63 (yy, 1 = 3.7 and 11.1 Hz, 1H), 3.49 (yy, 1 = 6.4 and 11.1 Hz, 1H), 2.3 (yy , 1 = 9.7 and 16.1 Hz, 1H), 1.77 (st, 1 = 1.9 and 16.0 Hz, 1H), 1.37 (t, 1H), 1.25 (t, 1H), 1.21 (t, 2H), 0.86 (t, 2H).

t / ζ 571 [M-1] ^- .

Biological activity under ίπ νίϋΌ conditions.

Example 92

Obtaining data 1C ₅₀ .

Materials and preparation of reagents. Human OBT-MEK1 and the constitutively active OBT-MEK1 ^CA allele (containing the Beg218Acr and Beg222Acr mutations) were subcloned into the yeast expression vector pOEM42 (Rgoteda, Maykop, XVI) from wild-type human MEK1 cDNA. OBT-MEK1 ^{CA was} expressed in Exxepsoy soy and partially purified using the Glutathione Sepharose 4B affinity polymer (Ategkyat Ryagtaya Vyuesy, Rksa1a ay. N1). The EKK2 allele was subcloned from the MAPK2 / EGK2 cDNA (wild-type) into the RIBEatr (Irk1a1e Vyuyesyoduod, 1 ps., Vayyat, MA) into the pET21a vector (No. vadep, Maikop, VI) to obtain the mouse EKK2 allele with the nth terminal. EKK2 was expressed and purified to homogeneity [2yaid, 1993 # 33]. The main myelin protein (MBP) was obtained from Oso VKI (Kosk \ z11e, MO). Eaku TMek adenosine 5'-triphosphate (ATP) ([у- ³³ Р]) (ΝΞΝ Regksh E1teg, Ve11ek1eu, MA) was the source of the radioactive label for all kinase reactions. Activated Ka £ -1 (truncated) and activated MAPKinase 2 / EKK2 were obtained from Irk1a1e, 1 ps. (Lake P1ac1y, ΝΥ). 4-20% Szhyepop Rgesak! gels were obtained from Vu-Kai (Negsiek, CA).

Determination of enzymatic activity. Compounds were diluted from stock solutions of dimethyl sulfoxide (DMSO) in 1xHM1MBE (20 mM NEREB pH 7.2, 1 mM MdCl ₂ , 100 mM No. C1, 1.25 mM ITT, 0.2 mM EITA). A typical 25 μl sample contained 0.002 nmol MEK1 ^CA , 0.02 nmol ECK2, 0.25 nmol MBP, 0.25 nmol unlabeled ATP and 0.1 μCi [at ³³ P] ATP. The screening study mainly included four additions. 5 μl of the diluted compound was dispersed in 96 well assay plates. Then, 10 μl of 2.5x enzyme cocktail (MEK1 and EKK2 only) was added to each well, after which they were preincubated for 30 min at ambient temperature. Then, 10 μl of a 2.5x substrate cocktail (labeled and unlabeled ATP plus MBP) was added, after which it was incubated for 60 min at ambient temperature. At the end, 100 was added.

- 48,020,624 μl of 10% trichloroacetic acid (TCA) and incubated for 30 min at room temperature to stop the reaction and precipitate radioactively labeled protein products. The reaction products were collected on 96-well filtered glass fiber plates pre-moistened with water and 1% pyrophosphate. After that, the filter plate was washed 5 times with water. Water was superseded by absolute ethanol and the plates were allowed to air dry for 30 minutes at room temperature. Manually sealed from the bottom and 40 μl of scintillation cocktail was dispersed in each well. Sealed from above and the plate was analyzed in TorSonSh for 2 s per well.

For certain experiments, a truncated version of MEK was used, for which activation of Ka! kinase.

Example 93

Obtaining EU data ₅₀ .

The actions of the compounds in the cell were determined using Western blotting for phosphorylated NKK. Breast cancer cells MEA-MV-231 were seeded in a 48-well plate at a density of 20 thousand cells per well and grown at 37 ° in a humidified CO ₂ incubator. The next day, growth medium (EMEM + 10% fetal bovine serum) was removed and replaced with lunch medium (EMEM + 0.1% fetal bovine serum). Cells were incubated in a depleted medium for sixteen hours and then treated with compounds at various concentrations for 30 minutes. After incubation with the compound, the cells were stimulated with 100 ng / ml EOR for 5 minutes. Cells were then lysed and analyzed by Western blotting using a monoclonal antibody with increased affinity for phosphorylated NKK. The signal was amplified using a secondary antibody conjugated with a dye in the near infrared region and analyzed on a Ysog Oyukkeu scanner. The signal intensity was quantified and these data were used to obtain dose response curves and calculate the EC ₅₀ .

Designation: A, EC ₅₀ = <2.0 nM; B, EC ₅₀ = 2.0-15 nM; C, EC ₅₀ = 15 nM-100 nM; Ό, EC ₅₀ > 100 nM, 1C ₅₀ <20 μM; P, EC ₅₀ > 100 nM, 1C ₅₀ > 20 μM.

Designation: A - EC ₅₀ = <2.0 nM; B - EC ₅₀ = 2.0-15 nM; C - EC ₅₀ = 15 nM-100 nM; Ό - EU ₅₀ = 100-200 nM; E - EC ₅₀ > 200 nM; BUT = not yet determined.

Biological activity in the conditions ίη У1уо.

Example 94

The compounds and compositions described herein are suitable for the treatment or prophylaxis of one or more diseases, including, but not limited to malignant neoplasm, inflammatory bowel disease (ΙΒΌ), psoriasis, and rheumatoid arthritis (CA). The compounds and compositions described in this invention are also suitable for oral administration once a day or twice a day for the treatment or prophylaxis of one or more diseases, including, but not limited to, malignant neoplasms, ΙΒΌ, psoriasis, and CA.

Testing the compound of the structure shown below (compound A, prepared as described in this invention) under ίη ugo conditions is described in this example

Human tumors were implanted with pi / pi mice. Compound A was administered orally for 14 days after the tumor size was about 100 mm ³ . Tumor growth inhibition (TO1) was determined after 14 days of treatment as a decrease in the size of the tumors in the treated groups relative to the control groups receiving the vehicle. The time to the control point (TTE) was calculated as the period of time during which the tumor reaches a certain final volume or the last day of the study, regardless of which comes first. The treatment outcome was determined based on the percentage of tumor growth retardation (% TSE). defined as an increase in the percentage of the average TTE of the treated animals relative to control mice that received the vehicle. Animals were also observed for regression of responses. The levels of rECK in the tumor and brain were determined using Western blotting and compared with the levels of compound A in the blood plasma for the study of pharmacodynamics / pharmacokinetics. The number of model tumors was evaluated for various doses and dosage regimens. Treatment with 25 or 50 mg / kg once a day (Οϋ) showed a statistically significant% TSE in A375 melanoma tumors, in Colo205 colon cancer, and A431 epidermoid tumors. Statistically significant TO1 was observed with the oral administration of doses of 25 mg / kg CE for these tumor models, as well as for cancer of the intestinal tunic HT29. The effect of various dosing regimens was evaluated on A375 xenografts.

- 49 020624

Although 100 mg / kg Compound A, once every two days, showed a statistically significant% ΤΟΌ (91%), it was not as effective as treatment CE at doses of 25 mg / kg (143% ΤΟΌ) or 50 mg / kg (233% ΤΟΌ). The dosage in the form of twice a day (W) was also more effective than the CE dosage, which was determined using% ΤΟΙ. A dosage of 12.5 mg / kg of HS was 79.5% ΤΟΙ compared to 51.7% for 25 mg / kg of CE of compound A. A dosage of 25 mg / kg of HS was 110.1% ΤΟΙ compared to 69.9 % ΤΟΙ for 50 mg / kg CE. A pharmacodynamic / pharmacokinetic study in Co1205 xenografts showed inhibition of pECK formation in tumors, while minimal inhibition was observed in the brain, indicating an effective antitumor activity with limited penetration into the central nervous system.

Compound A is an effective MEK1 / 2 inhibitor that inhibits the growth of клетокη νίίτο and ίίη νίνο tumor cells. SCAR determines susceptibility to growth inhibition by using a compound in anchor-dependent growth, but not in anchor-independent growth or in xenografts. It is believed that maintaining sufficient MEK inhibition for the entire dosing interval is more important than peak levels due to greater efficacy with more frequent dosing. Compound A has a favorable pk profile in humans, with an estimated therapeutic dose, based on xenograft studies, 20–40 mg / day in humans.

Example 94A

Inhibition of cancer cell growth (ΟΙ ₅₀ ).

Inhibition of anchor-dependent growth was assessed using a ΟοΙΙΤι чογΟΙο reagent after 48 hours of treatment with compound A for cell growth in 384 well wells. For studies of anchor independent growth, reagent ΜΤ8 (methanethiosulfonate) was used after treatment for 7 days of cell growth in a medium containing 0.15% agarose or on non-binding plates (A431). The growth inhibition values (ΟΙ ₅₀ ) are presented in the table below.

Tumor cell line SCAR status OB® anchoring dependent (nM ± s.o.) Eye Anchor Independent (nM ± s.o.) AZ 75 Melanoma U600E 67 ± 12 68 ± 34 Co1o205 Colon U600E 74 ± 45 33 ± 16 NT29 Colon U600E 70 ± 12 Not determined A431 Epidermoid Normal > 10,000 65 ± 19

Example 94B

Antitumor activity on xenografts.

Female pi / pi mice were implanted with A375 melanoma cells, Colo205 colon tumors, A431 epidermoid tumors, or HT-29 colon tumors, which were allowed to grow to 100,200 mm ³ . Compound A or excipient was administered orally (25, 50 or 100 mg / kg), once a day, for 14 days. The mean tumor volumes were plotted for filled and treated groups and depicted in FIG. one.

Example 94C.

Tumor growth inhibition ΤΟΙ 25 mg / kg CE.

Tumor growth inhibition for the treated groups for a dose of 25 mg / kg of compound A was calculated for these tumor xenografts. Inhibition of tumor growth was measured after dosing once a day for 14 days and calculated according to:

To1% = 100 x 1 - (volume of tumor treatment echnyi ^ - tumor volume ^ _{coagulant nyN)} (tumor volume of the treated filler _is finite - tumor volume _{mw | and | s)}

The range for the A375 and Co1205 represents values from 2 separate studies.

Xenograft tumors % TO1 R value AZ 75 Melanoma 52-72 ** <0.001 Co1o205 Colon 70-123 ** <0.001 NT29 Colon 56 <0.001 A431 Epidermoid 67 <0.001

** Noted regression during the experiment.

Example 94Ό.

EI ₅₀ in Co1205 xenografts.

Male pi / pi mice were implanted with Co1O205 tumor cells. After treatment for 10 days, the animals were randomized according to the size of the tumor (interval 126-256 mm ³ ) and treated with paclitaxel (Ιν, ΟΘΌ / 5). filler or compound A (PO, CE / 14).

Pharmacokinetic parameters were obtained by dosing Ba1b / c in mice with doses of 25 mg / kg of compound A and extrapolating the values for lower dose groups and the results are presented in the table below.

Group η Treatment regimen The initial tumor volume (mm ³ ) Tumor volume on day 15 (mm ³ ) % TO1 g '“LIKE (mcg / ml) C | gtsp (mcg / ml) foxes (mcg / h) Means mg / kg one 10 Will fill up b - 185 ± 11.1 2093 ± 174 - - - - 2 10 Paclitaxel thirty 184 ± 9.8 113 ± 9.6 104 * - - - 3 10 Compound BUT 2,5 184 ± 9.8 P87 ± 127 47 * 0.99 0.003 5.5 4 10 5 183.8 ± 9.8 P75 ± 104 48 * 1.97 0.006 11.0 5 10 10 185.1 ± 11.7 1045 ± 160 55 * 3.94 0.012 22.0 6 10 25 185.1 ± 11.7 762 ± 81 70 " 9.85 0,029 55.0

* P <0.001.

Example 94E

Inhibition of tumor growth with A375 xenografts.

Compound A 50 mg / kg А was administered to A375 xenograft mice. 25 mg / kg VGO, 50 mg / kg ΟΌ and 12.5 mg / kg VGO. % ΤΟI was calculated, presented graphically and depicted in FIG. 2.

Example 94P

Plasma concentrations in mice.

Female pi / pi mice were implanted with A375 tumor cells, which were allowed to grow to 100,200 mm ³ . Compound A or excipient was administered orally once a day (ΟΌ) or twice a day (VGO) (50 mg / kg ΟΌ, 25 mg / kg VGO, 50 mg / kg ΟΌ and 12.5 mg / kg VGO). Tumor growth inhibition was measured after dosing once a day for 14 days and calculated according to:

To1% = 100 x 1 - (amount of treatment of tumors _not-screw - volume op \ -holi _{npish, n,} n) (tumor volume of vehicle-treated _horse-screw - tumor volume _initial th)

AIS (μg h / ml) 132.5 117.0 66.5 78.0 S _tach (MKG / ML) 23.8 10,2 11.9 7.8 S _t1p (MKG / ML) 0.06 1.24 0,03 0.49 SST Free fractions (ng / ml) 0.117 2.48 0.059 0.986

Statistical significance = log-rank test.

Example 940

Xenograft tumors in mice and inhibition of MEK activity in the brain.

Female pi / pi mice that implanted Co1205 tumor cells were given a single dose of vehicle or compound A at 2.5, 5, 10 or 25 mg / kg. Concentrations of compounds were determined in plasma samples, and pEPK levels collected through tumor and brain samples

2, 6, 12, and 24 hours after dosing. RERK levels after western blotting were quantified using Y-COP Obukkeu, normalized to total EPK levels, and compared with vehicle levels to determine% MEK inhibition. MEK inhibition in a tumor or brain for each mouse was graphically represented with the corresponding plasma concentrations of compound A in the animal. Nonlinear regression gave EC ₅₀ 73 nM for inhibition of MEK in tumors. EC ₅₀ values in the brain were> 5000 nM.

A plot of plasma concentration (1ode nM) versus% inhibition of pEPK is shown in FIG.

3.

Preparation of capsules.

Example 95A

Prepared blue hard gelatin capsules of size 1, which contain a dry powdered powdered composition in doses of 1 and 10 mg of compound A (see table, presented in example 93 above) structure:

Compound A was prepared as described in this invention, and then micronized using a jet mill (8p1ga1 1e (MS, electronically milled, with a grinding chamber diameter of 50 mm; 50 °. 4x0.8 mm atomizer ring; nozzle atomizer diameter 0.8 mm and spray distance

- 51 020624 nozzles 3 mm). Compound A and a portion of microcrystalline cellulose were mixed and sieved through a No. 20 sieve and added to a diffuser drum grinder (Y grinder). The remaining microcrystalline cellulose was sieved through a No. 20 sieve, added to the substances in a grinder, and ground. Croscarmellose sodium and sodium lauryl sulfate were sieved through a No. 20 sieve, added to the substances in a grinder, and ground. The powder mixture was passed through a rotary impact-reflective mill (Ciabgo SoMP), added back to the grinder and continued to grind. Magnesium stearate was sieved through a No. 20 sieve and ground with a powder mixture obtained by grinding. The powder mixture was filled into size 1 capsules. A strip for identification was applied to 10 mg capsules.

The composition of the capsules is presented in the table below.

Component 1 mg capsule 10 mg capsule mg / unit % ig / unit % Compound A 1,0 0.4 10.0 4.2 Microcrystalline cellulose, ΝΡ (Ανίοβΐ PH302) 222.2 92.6 213.2 88.8 Croscarmellose sodium, ΝΡ (Ac-Οί-δοΙ) 12.0 5,0 12.0 5,0 Sodium lauryl sulfate, ΝΡ 2,4 1,0 2,4 1,0 Magnesium Stearate, ΝΡ 2,4 1,0 2,4 1,0 ³ total 240.0 100.0 240.0 100.0 Blue hard coated gelatin capsules size 1 one one

^{and The} target fill weight was adjusted based on the actual activity of the mixture.

The typical batch composition for 10 thousand units of 1 mg capsules was as follows.

Batch Composition Components Amount per lot (g) (for 10 thousand units) Compound A 10.0 Microcrystalline cellulose, ΝΡ (Ανί "1 PH302) 2222 Croscarmellose sodium, ΝΡ (Ac-ϋί5o1) 120.0 Sodium lauryl sulfate, ΝΡ 24.0 Magnesium Stearate, ΝΡ 24.0 Total weight ⁸ 2400 Blue hard coated gelatin capsules size 1 10 thousand

^{and The} target fill weight was adjusted based on the actual activity of the mixture.

The typical lot composition for 10 thousand units of 10 mg capsules was as follows.

Batch Composition Components Amount per lot (g) (for 10 thousand units) Compound A 100.0 Microcrystalline cellulose, ΝΡ (A \ '1cc1 PH302) 2132 Croscarmellose sodium, ΝΡ (Ac-ϋϊ8o1) 120.0 Sodium lauryl sulfate, ΝΡ 24.0 Magnesium Stearate, ΝΡ 24.0 Total weight * 2400 Blue hard gelatin capsules size 1 with a shell ¹ * 10 thousand

^{and The} target fill weight was adjusted based on the actual activity of the mixture.

Example 95B

Prepared solid blue gelatin capsules of size 1, which contain a dry powdered powdered composition in doses of 1 and 10 mg of compound B (see table, presented in example 93 above) structure:

- 52,020,624

Compound B was prepared as described in this invention and micronized using a jet mill (8p1ga1 Le1 MSH, electronically grinding, with a grinding chamber diameter of 50 mm; 50 °, 4x0.8 mm atomizer ring; nozzle atomizer diameter 0.8 mm and distance atomizer nozzle 3 mm). Compound B and a portion of microcrystalline cellulose were mixed, sieved through a No. 20 sieve, and added to a diffuser drum shredder (Y shredder). The remaining microcrystalline cellulose was sieved through a No. 20 sieve, added to the substances in a grinder, and ground. Croscarmellose sodium and sodium lauryl sulfate were sieved through a No. 20 sieve, added to the substances in a grinder, and ground. The powder mixture was passed through a rotary impact-reflector mill (Oiabgo CoMb), added back to the grinder and continued to grind. Magnesium stearate was sieved through a No. 20 sieve and ground with a powder mixture obtained by grinding. The powder mixture was filled into size 1 capsules. A strip for identification was applied to 10 mg capsules.

The composition of the capsules is presented in the table below.

Component 1 mg capsule 10 mg capsule mg / unit % ig / unit % Compound B 1,0 0.4 10.0 4.2 Microcrystalline cellulose, ΝΡ (A \ sce1 PH302) 222.2 92.6 213.2 88.8 Croscarmellose sodium, ΝΡ (Ac-ϋΐ-δοΐ) 12.0 5,0 12.0 5,0 Sodium lauryl sulfate, ΝΡ 2,4 1,0 2,4 1,0 Magnesium Stearate, ΝΡ 2,4 1,0 2,4 1,0 To1a1 " 240.0 100.0 240.0 100.0 Blue hard coated gelatin capsules size 1 one one

Activity in humans under the conditions ίη νί \ Ό.

Example 96

The introduction of the capsules described in example 95A, people with malignant tumors. Patients with malignant neoplasms were administered once in capsules at a dose of 1 or mg of the composition described above in Example 95A. For a dose of 2 mg, patients were given 2x 1 mg capsules; for a dose of 4 mg, patients were given 4x1 mg capsules; for a dose of 6 mg, patients were given 6x1 mg capsules; for a dose of 10 mg, patients were given 1x10 mg capsules; for a dose of 20 mg, patients were given 2x10 mg capsules.

The concentration profiles were observed versus time and the results are shown in FIG. 4 and in the table below.

Dose (mg) Day TSCH (h) Stah (ng / ml) C | 2b (ng / ml) AIS _th .12y (ng-h / ml) sheet (ng-h / ml) 2 one 2.0 0,111 0,0378 0.700 nd 35 2.0 0.202 0,0756 VD 2.07 4 one 1,5 0.292 0.134 2.26 nd 35 1,0 0.544 0.310 nd 5.12 10 35 nd 1,57 1.01 nd 14.3 twenty 35 nd 3.28 2.19 nd 29.5

Crystalline polymorphic forms.

Example 97

Preparation of N- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

^ (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) phenyl) -1 - (2,3-dihydroxypropyl) cyclopropan-1 sulfonamide was prepared according to the previously described methods (see published international patent application νθ 2007 / 014011) and as described below.

- 53,020,624

Stage A. 2-Fluoro ^ - (2,3,5-trifluoro-6-nitrophenyl) -4-iodobenzeneamine.

A solution of 1.0 M lithium hexamethyldisilazide (L ^ 81Me ₃ ) ₂ ) ΈΗΜΌ δ (15.37 ml, 15.37 mmol) was slowly added to the stirred solution of 2-fluoro-4-iodaniline (3.64 g, 15.37 mmol ) in anhydrous THF (100 ml) under nitrogen at -78 ° C and stirring continued at -78 ° C for another hour. 2,3,4,6-tetrafluoronitrobenzene was added, the reaction mixture was allowed to warm to room temperature and stirring was continued for an additional 16 hours. Ethyl acetate (200 ml) was added and the organic phase was washed with water, dried over sodium sulfate and further purified by column chromatography. receiving the product as a yellow solid (3.75 g, 59.24%).

MH +: 410.9.

Ή NMR (DMSO, 300 MHz): 6.85 (ΐ, 1Η); 7.38 (ά, 1Η); 7.62 (t, 2Η); 8.78 (5, 1Η).

Stage B. 2-Fluoro - ^ (2,3-difluoro-5-methoxy-6-nitrophenyl) -4-iodobenzeneamine.

A stirred solution of (2-fluoro-4-iodophenyl) - (2,3,5-trifluoro-6-nitrophenyl) amine (1.23 g, 3 mmol) in anhydrous THF (25 ml) in a nitrogen atmosphere was cooled to -78 ° C and a solution of 25% sodium methylate (0.68 ml, 0.3 mmol) was slowly added. The reaction mixture was allowed to warm to room temperature and continued to stir for an additional 16 hours. TLC indicated incomplete reaction. Ethyl acetate (100 ml) was added to the reaction mixture, and the organic layer was washed with water, dried over sodium sulfate and further purified by column chromatography to give the desired compound as a yellow solid (0.6 g, 47.6%).

t / ζ 424 [M + H] +.

Step C. 5,6-Difluoro - ^ - (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 solution (2,3-difluoro-5-methoxy-6-nitrophenyl) - (2- fluoro-4-iodophenyl) amine (0.57 g, 1.34 mmol) in ethanol (20 ml). The mixture was stirred under heating under reflux for 16 hours, cooled to room temperature, filtered through celite, and the filtrate was concentrated to dryness. The resulting residue was resuspended in ethyl acetate, washed with water, dried over sodium sulfate and further purified by crystallization from ethanol to give the product as an off-white solid (0.47 g, 90.3%).

M-H ⁺ : 393.2.

Ή NMR (DMSO, 300 MHz): 3.76 (5, 3Η); 6.1 (ΐ, 1Η); 6.8-7.0 (t, 1Η); 7.2 (ά, 1Η); 7.35 (5, 1Η); 7.42 (ά,

1Η).

Stage Ό. 1-Allyl - ^ (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-1sulfonamide.

To a stirred solution of 5,6-difluoro - ^ - (2-fluoro-4-iodophenyl) -3-methoxybenzene-1,2-diamine (1 equiv.) In anhydrous pyridine (5 ml / mmol) was added 1-allylcyclopropanesulfonyl chloride (1 -5 equiv.). 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 (Mg§ ₄ ) and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica to give the title product.

Ή NMR (СОС1 ₃ , 300 MHz): δ 7.417 (άά, 1Η), 7.309 (5, 1Η), 7.25 (t, 1Η), 6.89 (t, 1Η), 6.52 (t, 1Η ), 6.427 (t, 1Η), 6.03 (5, 1Η), 5.668 (t, 1Η), 5.11 (ΐ, 1Η), 3.9 (5, 3Η), 2.75 (ά, 2Η ), 1.21 (t, 2Η), 0.767 (t, 2Η).

Stage E. ^ (3,4-Difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

1-Allyl - ^ (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) cyclopropane-1-sulfonamide (97 mg, 0.18 mmol) and 4-methylmorpholine Ν-oxide ( 21 mg, 0.18 mmol) was dissolved in THF (8 ml). Osmium tetroxide was added at room temperature (0.018 mmol, 0.13 ml, 4% at Η20) and the reaction mixture was stirred at room temperature for 16 hours. Ethyl acetate was added, the organic phase was washed with water, dried (Mg§0 ₄ ) and concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluants: EuAc / MeOI). receiving the title product (0.80 g, 78%).

Ή NMR (СОС1 ₃ , 300 MHz): δ 7.38 (άά, 1 = 1.7 and 10.3 Hz, 1Η), 7.26 (t, 1Η), 7.14 (5, 1Η), 6 87 (5, 1Η), 6.53 (άά, 1 = 6.8 and 11.4 Hz, 1Η), 6.43 (t, 1Η), 4.06 (t, 1Η), 3.89 ( 5, 3Η), 3.63 (άά, 1 = 3.7 and 11.1 Hz, 1Η), 3.49 (άά, 1 = 6.4 and 11.1 Hz, 1Η), 2.3 (άά , 1 = 9.7 and 16.1 Hz, 1Η), 1.77 (άά, 1 = 1.9 and 16.0 Hz, 1Η), 1.37 (t, 1Η), 1.25 (t,

- 54,020,624

1H), 1.21 (t, 2H), 0.86 (t, 2H). t / ζ 571 [M-1] ^- .

Example 98

Preparation of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

R

The pure δ isomer was obtained by chiral HPLC separation of the racemic mixture.

Ή NMR (SIS1 ₃ , 300 MHz): δ 7.38 (bb, 4 = 1.7 and 10.3 Hz, 1H), 7.26 (t, 1H), 7.14 (s, 1H), 6 87 (h, 1H), 6.53 (bb, 1 = 6.8 and 11.4 Hz, 1H), 6.43 (t, 1H), 4.06 (t, 1H), 3.89 ( h, 3H), 3.63 (bb, 1 = 3.7 and 11.1 Hz, 1H), 3.49 (bb, 1 = 6.4 and 11.1 Hz, 1H), 2.3 (bb , 1 = 9.7 and 16.1 Hz, 1H), 1.77 (bb, 1 = 1.9 and 16.0 Hz, 1H), 1.37 (t, 1H), 1.25 (t, 1H), 1.21 (t, 2H), 0.86 (t, 2H).

t // 571 [M-1] ^- .

Example 99

Preparation of ^ (K) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide

Pure K isomer was obtained by chiral HPLC separation of the racemic mixture.

Ή NMR (SIS1 ₃ , 300 MHz): δ 7.38 (bb, 1 = 1.7 and 10.3 Hz, 1H), 7.26 (t, 1H), 7.14 (s, 1H), 6 87 (h, 1H), 6.53 (bb, 1 = 6.8 and 11.4 Hz, 1H), 6.43 (t, 1H), 4.06 (t, 1H), 3.89 ( h, 3H), 3.63 (bb, 1 = 3.7 and 11.1 Hz, 1H), 3.49 (bb, 1 = 6.4 and 11.1 Hz, 1H), 2.3 (bb , 1 = 9.7 and 16.1 Hz, 1H), 1.77 (bb, 1 = 1.9 and 16.0 Hz, 1H), 1.37 (t, 1H), 1.25 (t, 1H), 1.21 (t, 2H), 0.86 (t, 2H).

t // 571 [M-1] ^- .

Example 100

Preparation of the crystalline polymorphic form A ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Preparation ί) ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (216.10 g) loaded into a 4 L Erlenmeyer flask equipped with a large magnetic stirrer and a magnetic stirrer / heating plate. Ethyl acetate was added (about 600 ml obtained from Izb). Heating and stirring initiated the formation of a brown suspension. The mixture was brought to mild reflux and ethyl acetate (about 200 ml) was added to complete the dissolution to give a dark brown solution. Heptane (obtained from Asgoz) was slowly added in portions to the refluxed solution at a rate at which all the precipitated substances that formed with each addition were rapidly dissolved and reflux maintained. After adding 2 L of heptanes to the solution, the formed solids dissolved very slowly upon reflux. Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature with stirring for 16 hours. A thick layer of crystalline substance formed on the surface of the glass during the aging period. The resulting suspension was balanced in an ice / water bath with stirring. The suspension was filtered through a 25 cm Buchner funnel with Whatmann filtration material No. 1. The collected crystals were washed with heptanes (1 L) and allowed to air dry in vacuum. The crystals were additionally dried at 40 ° C / <1 torr for 20 hours to give the product as a pink crystalline solid (160.99 g, 77.2%).

Preparation ίί) No. (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (13.2 g) and ethyl acetate (30 ml) was charged into an Erlenmeyer flask equipped with a large magnetic stirrer and a magnetic stirrer / heating plate. Heating and stirring were initiated to mild reflux to complete the dissolution, resulting in a dark brown solution. Heptanes were slowly added in portions to the refluxed solution at a rate at which all the precipitated substances that formed with each addition were quickly dissolved and reflux maintained until the addition of heptanes to the solution caused the solids to dissolve very slowly upon reflux (~ 90 ml heptanes). Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature with stirring for 16 hours. A thick layer of crystalline material was formed on the surface of the glass during the aging period. The resulting suspension was balanced in an ice / water bath with stirring. The suspension was filtered through a Buchner funnel with Whatmann filtration material No. 1. The collected crystals were washed with heptanes and allowed to air dry in vacuum. The crystals were further dried at 40 ° C / <1 torr for 20 hours to give the product as a pink crystalline solid.

Preparation ίίί) ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1-sulfonamide (44.8 g) and ethyl acetate (750 ml) was charged into an Erlenmeyer flask equipped with a large magnetic stirrer and a magnetic stirrer / heating plate. Heating and stirring were initiated to mild reflux to complete the dissolution, resulting in a dark brown solution. Hexanes were added very slowly in portions to the refluxed solution at a rate at which all the precipitated substances that formed with each addition were rapidly dissolved and reflux maintained, until the addition of hexanes to the solution caused the solids to dissolve very slowly during reflux (~ 2 l of hexanes). Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature with stirring for 16 hours. A thick layer of crystalline substance formed on the surface of the glass during the aging period. The resulting suspension was balanced in an ice / water bath with stirring. The suspension was filtered through a Buchner funnel with Whatman No. 1 filtration material. The collected crystals were washed and allowed to air dry in vacuum. The crystals were further dried at 40 ° C / <1 torr for 20 hours to give the product as a pink crystalline solid.

Example 101

Preparation of crystalline polymorph ^ (K) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Ν- (Κ) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropan-1-sulfonamide (216.10 g) was loaded into 4 L Erlenmeyer flask equipped with a large magnetic stirrer and a magnetic stirrer / heating plate. Ethyl acetate (about 600 ml) was added. Heating and stirring initiated the formation of a brown suspension. The mixture was brought to mild reflux and ethyl acetate (about 200 ml) was added to complete the dissolution to give a dark brown solution. Heptane was slowly added in portions to the refluxed solution at a rate at which all the precipitated substances that formed with each addition were rapidly dissolved and reflux was maintained. After adding 2 l of heptanes to the solution, the formed solids dissolved very slowly upon reflux. Heating was stopped and the crystallization mixture was allowed to equilibrate to room temperature with stirring for 16 hours. A thick layer of crystalline substance formed on the surface of the glass during the aging period. The resulting suspension was balanced in an ice / water bath with stirring. The suspension was filtered through a 25 cm Buchner funnel with Whatmann filtration material No. 1. The collected crystals were washed with heptanes (1 L) and allowed to air dry in vacuum. The crystals were additionally dried at 40 ° C / <1 torr for 20 h.

Example 102

Obtaining data 1C ₅₀ .

Materials and preparation of reagents. O8T-human MEK1 and constitutively active MEK1 allele ^O8T-ca (containing mutations and §et218Akr §et222Akr) was subcloned into the yeast expression vector rOEM42 (Rgoteda, Mab1kop, XVI) MEK1 cDNA of human wild type. O8T-MEK1 ^{CA was} expressed in Excepsois and partially purified using the Glutathione Sepharose 4B affinity polymer (AtegkNat Pbattia B | o1ecH, Pcacpuau, S). The EKK2 allele was subcloned from the MAPK2 / EGK2 cDNA (wild-type) into pI8Eatr (Irk1a1e Vu1esbpododu, 1 ps., A11yat, MA) into the pET21a vector (No. uadep, Mab1kop, VI) to obtain the mouse eKkomin mouse allele. EKK2 was expressed and purified to homogeneity [2Lap, 1993 # 33]. The main myelin protein (MBP) was obtained from OLso VKV (Koskushe, MI). Eakuibek adenosine 5'-triphosphate (ATP) ([у- ³³ Р]) (ΝΉΝ Regksh E1teg, \ e11ek1eu, MA) was a source of a radioactive label for all kinase reactions. Activated Kak-1 (truncated) and activated MAPKinase 2 / EKK2 were obtained from irk1a1e, 1ps. (Bake Р1асШ, ΝΥ). 420% CgPepop Rgesak! gels were obtained from Vu-Kab (Netsi1e8, CA).

Determination of enzymatic activity. Compounds were diluted from stock solutions of dimethyl sulfoxide (DMSO) in 1x1 Ι \ 1ΝΌΗ (20 mM HEPE8 pH 7.2, 1 mM MdCl ₂ , 100 mM No. C1, 1.25 mM ITT, 0.2 mM EITA). A typical 25 μl sample contained 0.002 nmol MEK1 ^CA , 0.02 nmol ECK2, 0.25 nmol MBP, 0.25 nmol unlabeled ATP and 0.1 μCi [at ³³ P] ATP. The screening study mainly included four additions. 5 μl of the diluted compound was dispersed in 96 well assay plates. 10 μl of a 2.5x enzyme cocktail (MEK1 ^CA and EKK2 only) was then added to each well, after which they were preincubated for 30 min at ambient temperature. Then 10 μl of 2.5x substrate cocktail (labeled and unlabeled ATP plus MBP) was added, after

- 56,020,624 of this were incubated for 60 minutes at ambient temperature. Finally, 100 μl of 10% trichloroacetic acid (TCA) was added and incubated for 30 min at room temperature to stop the reaction and precipitate radioactively labeled protein products. The reaction products were collected on 96-well filtered glass fiber plates pre-moistened with water and 1% pyrophosphate. After that, the filter plate was washed 5 times with water. Water was superseded by absolute ethanol and the plates were allowed to air dry for 30 minutes at room temperature. Manually sealed from the bottom and 40 μl of scintillation cocktail was dispersed in each well. It was sealed from above and the plate was analyzed in TorCoIP for two seconds per well. For certain experiments, a truncated version of MEK was used, which requires activation of the CaT kinase.

Example 103

Obtaining EU data ₅₀ .

The actions of the compounds in the cell were determined using Western blotting for phosphorylated NKK. MOL-MB-231 breast cancer cells were plated on a 48-well plate at a density of 20 thousand cells per well and grown at 37 ° in a humidified CO ₂ incubator. The next day, growth medium (ΌΜΕΜ + 10% fetal bovine serum) was removed and replaced with lunch medium (ΌΜΕΜ + 0.1% fetal bovine serum). Cells were incubated in a depleted medium for 16 hours and then treated with compounds at various concentrations for 30 minutes. After incubation with the compound, the cells were stimulated with 100 ng / ml EOR for 5 minutes. Cells were then lysed and analyzed by Western blotting using a monoclonal antibody with increased affinity for phosphorylated NKK. The signal was amplified using a secondary antibody conjugated with a dye in the near infrared region and analyzed on a Shot Obuzzeu scanner. The signal intensity was quantified and these data were used to obtain dose-response curves and the calculation of EC ₅₀ .

Example 104

Activity data of the compounds.

The compounds described in examples 1, 2 and 3 were tested in the studies described above. The results are summarized in the table below (A, EC ₅₀ = <2.0 nM; B, EC ₅₀ = 2.0-15 nM):

Connection No. Structure Activity Etc. 97 (Racemic) "and* one BUT Etc. 98 (8 isomer) no.-g L i lhd R BUT Etc. 99 (K isomer) pH Α ι IN

Example 105

Data ΧΚΡΌ (X-ray powder diffraction).

HREC was carried out on a 1pe1 KhKO-3000 diffractometer equipped with a nonlinear position-sensitive detector with a 2Θ interval of 120 °. Real-time data were collected using Xi Ka irradiation at a resolution of 0.03 ° 2Θ. The voltage and current on the tube were set equal to 40 kV and 30 mA, respectively. The paintings were viewed in the range from 2.5 to 40 ° 2Θ to facilitate direct comparison of the paintings. Samples of (8) -Y- (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (synthesized as described herein invention) was prepared for analysis by packing them in thin-walled glass capillaries. Each capillary was moved to a goniometric head, which was transferred to the motor mode, to allow rotation of the capillary during measurement and recording of parameters. Samples were analyzed for 5 minutes. Tool calibration was performed daily using a silicon standard

- 57,020,624 for comparison. In FIG. 5 is a graph of a powder X-ray diffraction pattern (ΡΧΚΌ) ^ (8) - (3,4-difluoro-2 (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide Form A. FIG. 7 is a graph of a powder X-ray diffraction pattern (ΡΧΚΌ) ^ (8) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide form A (top) and amorphous (bottom).

Example 106

Differential Scanning Calorimetry (E8C).

Analyzes were carried out on a TA Piitepei differential scanning calorimeter 01000. The instrument was calibrated using indium as a comparative substance. The sample was placed on a standard aluminum E8C cuvette with a non-corrugated lid configuration and the weight was accurately recorded. To determine the glass transition temperature (T _d ) of an amorphous substance, the cell with the sample was cyclized several times in the range from -40 to 140 ° C. The final temperature was brought to 150 ° C. T _{d was} recorded from the inflection point from the last transition cycle. In FIG. 6 is a graph of O8C modulated thermogram ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (form A ) The graph shows the normalized heat flux in Watts / gram (W / g) relative to the measured sample temperature in ° C.

Example 107

Sorption / desorption of dynamic evaporation (EU8).

Moisture sorption / desorption data were collected on a UT1 8CA-100 dynamic evaporation analyzer. Sorption and desorption data were collected in the range from 5 to 95% relative humidity (RH) at intervals of 10% RH under nitrogen purging. Samples were not dried before analysis. The equilibrium criterion used for the analysis was less than 0.0100% of the weight change in 5 minutes, with a maximum equilibration time of 3 hours, if the weight criterion was not met. The data were not corrected for the initial moisture content of the samples. Sodium chloride and polyvinylpyrrolidine were used as calibration standards. In FIG. Figure 8 shows the EU8 isotherm ^ (8) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (Form A). The substance shows an insignificant change in weight during the experiment.

Example 108

Thermogravimetry (TS).

Analyzes were performed on a TA 1n51gitep1 2950 thermogravimetric analyzer. Nickel and A1ite1 ™ were the calibration standards. Each sample was placed on an aluminum sample cuvette and inserted into a TC furnace. The samples were equalized at 25 ° C and then heated in a stream of nitrogen at a heating rate of 10 ° C / min, up to a final temperature of 350 ° C. In FIG. 9 shows the TS thermogram Ν (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1sulfonamide (form A), showing insignificant weight loss up to 140 ° C, indicating that polymorph Form A is unsolvated.

Example 109

Screening for malignant neoplasm in conditions ίη νίίτο.

Human tumor cell lines were grown in KPM11640 medium containing 5% fetal bovine serum and 2 mM L-glutamine. Cells were inoculated into microtiter plates of 96 wells in 100 μl with a seeding density in the range of 5 to 40 thousand cells / well, depending on the time of doubling of individual cell lines. After cell inoculation, microtiter plates were incubated at 37 ° C, 5% CO ₂ , 95% air and 100% relative humidity for 24 hours before adding ^ (8) - (3,4-difluoro-2- (2-fluoro- 4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

After 24 hours, two tablets of each cell line were fixed with ίη δίΐιι using TCA to allow measurement of the cell population for each cell line during the addition of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) ) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (T /). ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide was solubilized in dimethyl sulfoxide at 400 times the desired final maximum tested concentrations and kept frozen before use. At the time of addition, an aliquot of the frozen concentrate was thawed and diluted twice, preferably the final maximum test concentration, with a complete medium containing 50 μg / ml gentamicin. Additionally prepared 4-, 10-fold or _^half 1od serial dilution for a total of five concentrations plus control. Aliquots of 100 μl of these various dilutions were added to suitable microtiter wells already containing 100 μl of medium to obtain the desired final concentration.

After addition of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, the plates were further incubated for 48 hours at 37 ° С, 5% СО2, 95% air and 100% relative humidity. For attached cells, the study was stopped by adding cold TCA. Cells were fixed ίη δίΐιι by carefully adding 50 μl of cold 50% (w / v) TCA (final concentration, 10% TCA) and incubated in

- 58 020624 for 60 minutes at 4 ° C. The supernatant was discarded, the plates were washed five times with tap water and dried in air. Sulphorhodamine B (8KB) solution (100 μl) at 0.4% (w / v) in 1% acetic acid was added to each well and the plates were incubated for 10 minutes at room temperature. After staining, the unbound dye was removed by washing five times with 1% acetic acid and the plates were dried in air. The bound spot was then solubilized with 10 mM Trism base and absorbance was analyzed on an automatic plate reader at a wavelength of 515 nm. The methodology for the cell suspension was the same, except that the study was stopped by fixing the settled cells on the bottom of the wells by carefully adding 50 μl of 80% TCA (final concentration, 16% TCA). Using seven measurements of absorption [time zero (Τζ), control growth (C) and test growth in the presence of ^ (8) - (3,4-difluoro-2 (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) - 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide at five concentration levels (Τί)], the growth percentage was calculated for each drug concentration level. Percentage inhibition was calculated as:

Growth inhibition in percent = (Τϊ-Τζ) x 100 (concentrations for which Τί> / = Τζ) (С-Τζ)

Growth inhibition in percent = (Γί-Ί'ζ) x 100 (concentrations for which Τί <Τζ) Τζ

Three parameters of the dose-dependent effect were calculated. 50% growth inhibition (O1 ₅₀ ) was calculated from [(Τί-Τζ) / (Ο-Τζ)] χ 100 = 50, which is the concentration resulting in a 50% decrease in the increase in pure protein (as measured by 8KV staining) in control cells when incubated with the drug. Concentrations ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, leading to total growth inhibition (TO1 ), calculated from Τί = Τζ. BC ₅₀ (drug concentration leading to a 50% decrease in measured protein after drug treatment is over compared to that at the beginning of the study), indicating net cell loss after treatment calculated from [(Τ ^ -Τζ) / Τζ ] χ 100 = -50. Values were calculated for each of these three parameters if activity levels were reached; however, if the effect was not achieved or exceeded, then the values for the parameter were expressed as more or less than the maximum or minimum concentration tested.

The panels corresponding to leukemia, non-small cell lung cancer, colon cancer, CNS malignancy, melanoma, ovarian cancer, kidney cancer, prostate cancer and breast cancer were examined for the indicated cell lines and the results are presented below.

Panel Cell line 0150 (μm) Bc50 (μm) ΤΟΙ (μm) Leukemia SSKR-SEM 17,378 100,000 60,256 Leukemia NEWBO (TV) 0.010 100,000 100,000 Leukemia K-562 6,607 100,000 100,000 Leukemia MOLT-4 10,965 100,000 69,183 Leukemia ΚΡΜΙ-8226 26,915 100,000 100,000 Leukemia 5K 38,019 100,000 100,000 Non-small cell lung cancer A549 / ATCC 0.589 100,000 64,565 Non-small cell lung cancer ECUH 0.214 61,660 13,804 Non-small cell lung cancer NOR-62 0,069 42,658 12,589 Non-small cell lung cancer NOR-92 0,047 58,884 0.324 Non-small cell lung cancer N € 1-11226 3,311 74,131 24,547 Non-small cell lung cancer KS1-N23 0.056 74,131 2,884 Non-small cell lung cancer ΝΟΙ-Η322Μ 0.162 46,774 15,488 Non-small cell lung cancer Ν01-Η460 3,631 52,481 19,498 Non-small cell lung cancer ΝΟΙ-Η522 5,248 100,000 29,512 Colon cancer NSS-2998 0.010 0.457 0,035 Colon cancer NST-116 0.195 67,608 12,589 Colon cancer HCT-15 0.603 60,256 16,982

- 59,020,624

Colon cancer NT29 0,026 29,512 3,090 Colon cancer KM12 0.229 48,978 13,490 Colon cancer 8 \ ν-620 0,039 66,069 12,589 CNS malignant neoplasm 8R-268 2,570 100,000 25,704 CNS malignant neoplasm 8P-295 9,333 53,703 23,442 CNS malignant neoplasm 8R-539 1,514 60,256 20,417 CNS malignant neoplasm 8ΝΒ-19 0.251 75,858 24,547 CNS malignant neoplasm 8ΝΒ-75 0.302 34,674 4,467 CNS malignant neoplasm i251 0.891 44,668 17,378 Melanoma Lox 1MU1 0.195 38,905 10,715 Melanoma MAMME-ZM 0.010 19,953 0.014 Melanoma M14 0.015 29,512 0.166 Melanoma ZK-ME-28 0,028 22,387 0.214 Melanoma ZK-ME-5 0,062 38,905 13,804 Melanoma IACC-257 0,020 66,069 10,233 Melanoma ilss-62 0.014 20,893 0.170 Ovarian cancer ιοκονι 0.018 19,055 0.295 Ovarian cancer OUSAK-Z 2,512 48,978 17,783 Ovarian cancer OU SAK-4 0.562 72,444 16,218 Ovarian cancer OUSAK-5 0.017 40,738 12,023 Ovarian cancer 8K-OU-3 12,882 100,000 41,687 Kidney cancer 786-0 5,129 63,096 23,442 Kidney cancer A498 0.191 44,668 4,169 Kidney cancer ASN1CH 0.275 83,176 21,878 Kidney cancer SAKI 0.389 100,000 26,915 Kidney cancer 8Y12S 0.851 47,863 18,621 Kidney cancer TK-10 0.224 100,000 23,442 Kidney cancer io-31 0.158 40,738 11,482 Prostate cancer RS-Z 8,128 100,000 37,154 Prostate cancer ϋυ-145 2,138 95,499 22,387 Mammary cancer MCP7 10,965 85,114 30,903 Mammary cancer ΝΟΙ / ΑΟΚ- KE8 3,467 100,000 25,704 Mammary cancer ΜϋΑ-ΜΒ- 231 0,069 35,481 10,471 Mammary cancer NC 578T 0.617 85,114 13,490 Mammary cancer ΜϋΑ-ΜΒ- 0,035 41,687 12.303 435 Mammary cancer VT-549 5,754 47,863 20,893 Mammary cancer Τ-47ϋ 4,898 100,000 38,019 Mammary cancer ΜϋΑ-ΜΒ- 468 0.019 54,954 10,233

Example 110

Antiproliferative activity ίη νίΐΓΟ.

In this example, the following effects of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide were investigated: (1 ) activity (O1 ₅₀ ) in relation to several lines of tumor cells carrying various mutations; (2) activity (O1 ₅₀ ) with respect to the growth of several mutant B-CaG cell lines; (3) effects on anchor-independent cell growth; (4) effects on the cell cycle; and (5) toxic effects on the primary cells of the liver and kidneys.

Cell culture / growth inhibition assay.

Human A375 melanoma cells and human Colo205 colon cancer cells were obtained from ATCC (Mapa55a5, UA). A375 cells were maintained in IMEM supplemented with 10% fetal bovine serum, glutamine (2 mM), penicillin (100 units / ml) and streptomycin (100 μg / ml). Cells were maintained at 37 ° C, 5% CO ₂ and 100% humidity. Co1205 cells were maintained in CRM1 supplemented with 10% fetal bovine serum, glutamine (2 mM), penicillin (100 units / ml) and streptomycin (100 μg / ml). For experiments on growth inhibition, cells were placed in 384 well white microtiter plates at 1000 cells / 20 μl / well. After 24 hours, 5 μl of a 5X drug stock solution was added. All drugs were initially prepared in the form of 200X stock solutions in DMSO so that the final concentration of DMSO was 0.5%. Cells

- 60,020,624 were incubated for 48 hours at 37 ° C and ATP levels were determined using Ce11TyegO1o (Rgoteda, Maykop, XVI). The release of adenylate cyclase (AK) was determined using ToxPTSC (Satbxe, Xaahekegku1e, MO). Nonlinear processing of the curves was carried out using OgarRay Rpkt 4 (OgarRay 8th ^ ake, 8ap I1edo, CA). 4-amino-8 - ((2K, 3K, 48.5K) -3,4-dihydroxy-5- (hydroxymethyl) tetrahydrofuran-2yl) pyrido [2,3-th] pyrimidin-5 (8H) -one (UKH -14686) was a cytotoxic agent used as a comparative compound.

Growth Inhibition (%) = (Only Filler Control (CI) - L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl ) cyclopropane-1-sulfonamide KI) / (Filler control only KI-1 μM S- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 (2 , 3-dihydroxypropyl) cyclopropane-1-sulfonamide KI); based on growth arrest induced by Y- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, where measured ATP levels.

Cell viability (%) = (L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide Ki-10 μm UKH-14686 KII) / (Only Filler control Ki-10 μm Tamoxifen KII); based on cell lysis induced by UKX14686, where ATP levels were measured.

Cell lysis (%) = (L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide KII - Only filler control KII) / (10 μM Tamoxifen KII - Only filler control KII); based on cell lysis induced by tamoxifen where AK release was measured.

KII = Relative fluorescence units.

Assessment of cell cycle arrest.

A375 cells were placed in 96-well microplates at 10,000 cells / 200 μl / well. After 24 hours, cells reached a fusion of approximately 50% and 50 μl of a 5X stock solution of the drug was added. After another 24 h, the cells were trypsinized, fixed in 200 μl of PrheGeg (Apaec H, Va1e Sgeek, M1), and stored at 4 ° C overnight. Then the cells were washed in PB8, permealized, stained with 0.1% Triton X-100, 200 μg / ml RNAse-free DNase and 25 μg / ml propidium iodide (Mo1ci1ag Prgoec, 8ppuua, CA) and analyzed for Oiaua PCA-96 (Oiaua Teklnodlok, Rokljeg Syu, CA). The data were analyzed using MoiRb (version 3.0, Ugyu, Torquyat, ME).

(1) Assessment of inhibition of anchored cell growth.

The ultralow binding plate wells (Sogtpd, Ac1op MA) were filled with 60 μl of a 0.15% agarose solution in complete KPM1. Then, 60 μl of complete KPM1 containing 9000 Co205 cells in 0.15% agarose was added to the well. After 24 hours, 60 μl of a 3X drug solution in complete KPM1 without agarose was added. After 7 days, 36 μl of 6X MT8 reagent (Ce11Tyeg 96 Aceioik, Rgoteda, Maykop, VI) was added to the wells. After 2 hours at 37 ° C, the absorbance at 490 nm was determined on an M5 plate reader (Mo1eci1aGeU1sec, 8ppuuae, CA). Non-linear processing of the curves was carried out using OgaryRay Rpkt 4.

(2) Inhibition of growth (O1 ₅₀ ) relative to the MEK-dependent growth of cancer cells.

Mutant A375 mutant cells (human melanoma) dividing in the phase of logarithmic growth of B-CaG

A431 (melanoma), Co1205 (colon cancer), HT29 (colon and rectal adenocarcinoma), MEA-MB231 (breast adenocarcinoma) and BxRS3 (pancreatic adenocarcinoma) were exposed to X- (8) - (3,4-difluorine -2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide for 48 hours and the ATP content was analyzed. 100% growth arrest was determined using 1 μM L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1 - sulfonamide.

The table below presents the average O1 ₅₀ values of at least three experiments for each cell line and shows that N- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide causes growth inhibition in three B-CaH mutant cell lines (A375, Co1o205 and HT29), as well as in the same signaling pathway of hack / gaH / MEK / MAPK wild cell line type (A431) with an average efficiency of 79 nM (± 9 nM).

Line cells Average value Coeff. Student's Koeff variations A375 71 nM 12.1 nM 17% A431 86 nM 25.4 nM thirty% Co1o205 89 nM 40.1 nM 45% NT29 70 nM 12.2 nM eighteen% ΜϋΑ > 1 μM Arms > 1 μM

In a separate study, dividing in the phase of logarithmic growth of B-CaG mutant cells A375 (human melanoma), 8K Me128 (human melanoma) and Co1O205 (human colon cancer) were exposed to L- (8) - (3,4-difluoro-2 - (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide for 48 hours and the ATP content was examined. The table below presents O1 ₅₀ for each cell line, indicating that Ν- (8) - (3,4- 61 020624 difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1sulfonamide causes growth inhibition with an efficiency of approximately its EC ₅₀ value for MEK inhibition.

Cell line ΟΙ ₅ ο (ηΜ) A375 56 8K Me! 28 105 Co1o205 27

In FIG. 10A and 10B show the arrest of growth of dividing in the phase of logarithmic growth of A375 cells exposed to increased concentrations of ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) 6-methoxyphenyl) -1- (2 , 3-dihydroxypropyl) cyclopropane-1-sulfonamide. Cells were analyzed for ATP content. 100% growth arrest was determined using 1 μM ^ (§) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide .

In cell supernatants, cytotoxic lysis was analyzed by measuring the release of adenylate kinase (AK). The A375 dividing cells in the logarithmic growth phase were exposed to ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide and RE-325901 for 48 hours (100% of lysed cells were determined using 20 μM tamoxifen.) The results are shown in FIG. 11. These data indicate that ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide causes non-toxic growth arrest in some sensitive human cancer cell lines, as evidenced by ί) measuring growth arrest (ATP quantification); and ίί) lack of cytotoxic cell lysis (AK release). The absence of AK release was confirmed for all tested cell lines.

Inhibition of anchor independent growth.

Quantification of anchor-independent growth of Co1o205, A375, and MEAMB231 cells exposed to ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 (2,3- dihydroxypropyl) cyclopropane-1-sulfonamide for 7 days, in a 96-well microplate format. Viability was determined using an MT8 study. C1 ₅₀ values are presented below.

Cell line Average value Coeff. Student's Coeff. variations Co1o205 40 nM 8.1 nM twenty% AZ 75 84 nM 17.2 nM 21% ΜΩΑ- MV231 81 nM 55.6 nM 69%

In FIG. 12A-12C show growth inhibition of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (A) human Colon and colorectal cancer cells (C1 ₅₀ = 11 nM); (B) A375 cells (C1 ₅₀ = 22 nM); and (C) inhibition of MEA-MB231 cells that do not show indicated ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide growth arrest in two-dimensional studies dependent on anchoring.

The cells dividing in the logarithmic growth phase of A375 were exposed to Ν- (δ) - (3,4difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1sulfonamide ( 1 μM) for 48 h and growth inhibition (ATP content) and cytotoxic lysis (AK release) were analyzed in cell supernatants. 100% viability (ATP study) was determined in the vehicle only control wells. The table below shows the results indicating that ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1 β-sulfonamide causes non-toxic growth arrest in B-CaH mutant cells of human melanoma A375.

% control ATP, Cell Viability 27% AK, Cell Lysis 4%

Inhibition of anchor independent growth.

Anchor independent growth was quantified in 96-well microplate format. In FIG. 13A shows the inhibition of growth of human colorectal carcinoma cell So1o205 ₅₀ with values C1 to 6 concentrations and 11 nM, respectively. In FIG. 13B shows inhibition of growth of A375 cells with C1 ₅₀ values for concentrations of 5 and 22 nM.

The study of the cell cycle with induced ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide growth arrest.

MEK inhibition has been shown to induce cell cycle arrest in phase C1 / 8 on

- 62,020,624

A375 cells.

The cells dividing in the logarithmic growth phase of A375 were exposed to Ν- (8) - (3,4difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1sulfonamide for 24 hours and the percentage of cells that were stained in phase-dependent amounts of intracellular DNA was determined by flow cytometry.

The table below shows the percentage distribution of cells in the corresponding growth phases in cells treated with ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl ) cyclopropane-1-sulfonamidomy control (excipient only).

Phase% O1 8 02 The control 61.8 27.1 P, 1 N- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3- 111 nM 84.7 11.8 3,5 dihydroxypropyl) cyclopropane-1 sulfonamide 37 nM 74.3 18.7 7.0

In FIG. 14A and 14B show the effect of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on the cell cycle , indicating that the effect on the cells of A375 ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1- sulfonamide causes a stop in phase 01 of the cell cycle, as indicated by depletion in both phases 02 and 8.

Assessment of toxicity to primary hepatocytes and kidney cells.

Cryopreserved liver hepatocytes were obtained from CeP / Bpc! (Aikyp, TX) and were seeded onto 96-well collagen-coated plates according to the manufacturer's instructions. The drug was added 4 after sowing (final concentration of DMSO 0.5%).

Inoculated human hepatocytes received CeP / Bpc! and processed according to the manufacturer’s instructions.

Cryopreserved human kidney proximal tubule epithelial cells (CRTEC) were obtained from Satex and processed according to the manufacturer's instructions. Cells were grown for 4 days and then plated on 96-well plates at a density of 50 thousand cells / well for drug exposure.

After 48 h, the levels of AK in the supernatant were determined using ToxBrD, and the levels of ATP in the cells were determined using Ce11Tpeg01o. Total lysis values were determined using 15 μM UVX-14686.

The results are presented below. A very small number of lysed cells were observed. Minimal toxicity (81% survival) was observed at 30 μM ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1 -sulfonamide in freshly seeded primary human hepatocytes. For CRTEC, dose-dependent ATP depletion and apparent cell lysis at 30 μM were observed.

ATP (% cell survival) AK release (% cell survival)

Hepatocytes KRTES Hepatocytes KRTES

Compound A Rat Man Man Rat Man Man 30,0 57% 81% 34% 91% 109% 41% 10.0 72% 107% 85% 93% 112% 99% 33 87% 104% 91% 97% 102% 95% 1,1 114% 108% 94% 96% 92% 96%

The data presented above indicate that (1) ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane -1-sulfonamide inhibits cell growth and cell division in selected human cancer cells with O1 ₅₀ values in the range of 70-89 nM in studies of anchored proliferation without causing toxicity, as determined by analysis of cell lysis; (2) ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 (2,3-dihydroxypropyl) cyclopropan-1-sulfonamide inhibits cell growth and division in selected human cancer cells with O1 ₅₀ values of 51 and 22 nM in studies of proliferation dependent and anchor independent, respectively; (3) ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide causes stop 01 and inhibits the independent from anchoring growth in A375 cells, providing confirmation of anticancer activity in a physiologically relevant model under the conditions of the first; and (4) ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1-sulfonamide shows slight cytotoxicity with respect to in primary normal human hepatocytes, epithelial cells of the proximal tubules of the human kidney and kidney hepatocytes.

Example 111

Pharmacokinetic characteristics ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide in patients with malignant - 63 020624 neoplasms after multiple doses.

House (mg) N T _and „(h) (mcg / ml) Сз4) |, (mcg / ml) fox _t (mcg / h) * Ι / Ϊ (h) Stah dis _t 2 3 1.33 (21.7) 0,0504 (49.2) 0.00938 (82.8) 0.517 (61.2) 11,4 (38.8) 1.76 (35.6) 1.90 (23.9) 4 3 1,50 (33.3) 0.105 (41.0) 0,0313 (41.1) 1.39 (42.7) 14.9 (0,992) 1.49 (21.6) 1.91 (36.1) 6 3 1,50 (33.3) 0.205 (16.6) 0,0489 (12.2) 2.22 (5.79) 15.6 (23.8) 1,58 (38.5) 2.07 (23.5)

K _ac : accumulation index.

* Inaccurate assessment due to limited observation time.

After administration of multiple doses of ^ (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide to a subject in concentrations of 2, 4 or 6 mg ^ (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide is rapidly absorbed with medium the value of T _max in the range from 1.33 to 1.50 hours. The average values of C _max , C _t and AIS increase in proportion to the increase in dose. The accumulation indices are in the range from 1.49 to 1.76 for C _max and from 1.90 to 2.07 for AIS, respectively, which indicates moderate accumulation. Although the half-life cannot be accurately measured due to the limited observation time after multiple doses, it is believed that the half-life exceeds 22 hours after multiple doses, based on the accumulation indices. These half-life values are significantly higher than those observed on the efficacy model in mice, which are usually in the range of 2-3 hours. Additionally, encouraging ratios of peak to minimum values are observed for all doses.

Example 112

Pharmacokinetic characteristics ^ (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide in healthy volunteers after multiple doses .

Dose (mg) N t _mk (h) C ™ (mcg / ml) C, 4 Ig (μg / ml) fox (mcg / ml) »| / (h) Stah ^ W dis _t 10 6 2.00 (61.2) 0.182 (35.5) 0,0318 (53.0) 1.02 (1.80) (43.7) (39.7) 14.6 (15.2) 1.14 (19.0) 1.29 (13.4) twenty 6 2.25 (39.1) 0.313 (17.6) 0,0350 (36.5) 2.60 (22.0) 13,4 (21.9) 1.23 (24.1) 1.24 (6.51)

K _ac : accumulation index.

* Inaccurate assessment due to limited observation time.

After multiple doses of ^ (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide have been administered to a subject in concentrations of 10 or 20 mg ^ (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide is rapidly absorbed with an average value of T _max in the range from 2.00 to 2.25 hours. The average values of Stax, C _t and AIS increase with increasing dose. The accumulation indices are in the range from 1.14 to 1.23 for C _max and from 1.24 to 1.29 for AIS, respectively, which indicates insignificant accumulation. The half-lives are similar to those observed with the use of a two-dose regimen and range from 13 to 15 hours. These half-lives are shorter than those observed in patients with malignant neoplasm.

Example 113

Antiproliferative activity in w \% go.

The effect of ^ (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on the inhibition of cell proliferation was studied in the cell line originating from a cancer of a human stomach (stomach cancer) in a study of cell proliferation.

Cell culture / growth inhibition assay.

The human gastric cancer tumor cells Hk7461 were obtained from ATCC (Mapakkak, UA). Hk7461 cells were maintained in EMEM supplemented with 10% fetal bovine serum, penicillin (100 units / ml) and streptomycin (100 μg / ml). Cells were maintained at 37 ° C, 5% CO ₂ and 100% humidity. For studies of cell proliferation, cells were seeded in 96-well plates with a transparent bottom at a density of 3000 cells / 100 μl / well. After 24 hours, the cell medium was removed and replaced with a medium containing No. (B) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1 Sulfonamide in various doses. After incubation for 48 h at 37 ° C, ATP levels were determined using Ce11TIgC1o (Rgoteda, Maykop, VI) and the luminescence values were analyzed using Lb Vukuketk Apaukuk NT (Bipt a1e, CA). ATP levels for each

- 64,020,624 doses were determined in triplicate using independent wells.

Relative cell number = (average value of CI (treated with N- (3) - (3,4-difluoro-2- (2-fluoro4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropan-1 -sulfonamide)) / (average value of KI Only Filler control).

In FIG. Figure 19 shows a graph of cell numbers (relative to vehicle) versus Ν- (3) (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide and the obtained data indicate that N- (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane- 1-sulfonamide inhibits the proliferation of human gastric cancer cells Hk746! after 48 hours of treatment.

Example 114

Antiproliferative activity under conditions of shugo.

The effect of N- (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide on the inhibition of cell proliferation was studied in cell a line originating from stomach adenocarcinoma of the stomach (gastric cancer) in the study of cell proliferation.

Cell culture assay / growth inhibition.

Human gastric adenocarcinoma cells ASA were obtained from ATCC (Mapakkak, UA). ASA cells were maintained in EMEM / P12 supplemented with 10% fetal bovine serum, penicillin (100 units / ml) and streptomycin (100 μg / ml). Cells were maintained at 37 ° C, 5% CO ₂ and 100% humidity. For cell proliferation studies, cells were seeded in 96-well white plates at a density of 3000 cells / 100 μl / well. After 24 hours, the cell medium was removed and replaced with a medium containing Ν- (3) (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane -1 sulfonamide in various doses. After incubation for 3 days at 37 ° C, ATP levels were determined using Ce11TbegC1o (Ggoteda, Mabiop, XVI) and luminescence values were analyzed using Lb Vukuyetka Apauk1 NT (Zippuuae, CA). ATP levels for each dose were determined in triplicate using independent wells. In another experiment, 1000 cells / 100 μl / well were seeded and the cells were treated for 6 days and analyzed as described above.

Relative cell number = (average value of CI (treated with N- (3) - (3,4-difluoro-2- (2-fluoro4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropan-1 -sulfonamide)) / (average value of KI Only Filler control).

In FIG. 15A and 15B show a graph of the concentration of N- (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide the number of cells (relative to the vehicle) after (A) 3 days and (B) 6 days exposure to N- (3) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- ( 2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, which indicate that N- (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 ( 2,3-dihydroxypropyl) cyclopropane-1-sulfonamide inhibits the proliferation of human gastric adenocarcinoma cell line sheep NEA.

Example 115

Growth responses of orthotopic human Hep3B tumors in No. 1be mice treated with various amounts of N- (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2.3 dihydroxypropyl) cyclopropane-1-sulfonamide.

Dose-dependent response upon administration of N- (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (compound A) , regarding the inhibition of orthotopic human liver cancer, Hep3B2.1-7 was evaluated on BABB / c pi / pi mice, compared with the optimal dose of 5-fluorouracil (75 mg / kg).

Animals. For the study, female BABB / c pi / pi mice were used (Ishuegku o £ AbeYbe, \ νηί ^ Satrik, ZA, AikyPa), aged 10-14 weeks, with a body weight in the range of 19.1-29.94 g (average value 22.95 g). Mice were divided into 6 experimental groups (4 treated groups and 2 control groups) as follows.

The number of mice per group: 10 in groups from 1 to 5 inclusive; 15 in the control group of the reception rate (group 6).

The mice were kept in an environment with controlled conditions (target ranges: temperature 21 ± 3 ° C, humidity 30-70%, air exchange rate 10-15) in isolated conditions (quarantine) with a cycle of 12 hours of light / 12 hours of darkness. Temperature and relative humidity were monitored continuously. The animals were given commercially available food for rodents (Ka! Apb Moike Cieb, Zres1a1bu Reebk Ρίu Nb, C1ep Roggek !, Veket AikyaPa) and tap water ab Nbbit. Food and water were sterilized by autoclaving.

Inoculation of the tumor.

Hep3B cells of human liver cancer (passage 2 from the UT-ZYusk 353 working stock) were cultured in KGM11640 cell culture medium supplemented with 10% RHB and penicillin streptomycin (final concentration 50 IU / ml). Cells were harvested by trypsinization, washed twice in a refiner, and counted. After this, the cells were resuspended in a refiner: Ma1pde1 (1: 1, v / v)

- 65,020,624 and adjusted to a final volume containing 1/10 ⁸ cells / ml. Before inoculation, the incision sites were thoroughly washed with alcohol and the incision was made through the abdominal wall to access the liver. The needle was inserted into the liver cavity, where 10 μl of cells (1/10 ⁶ cells) were injected. The needle was held in this position for about 30 seconds to allow the polymerization of Ma (T1 §1® to prevent the leakage of tumor cells into the abdominal cavity.

Treatment began 14 days after inoculation. On the 7th day of the study (21 days after inoculation) all mice from the control group, the absorption rate was rejected and the liver was visually examined to determine the presence of a tumor.

Materials The following substances were obtained from appropriate commercial sources.

A sterile saline solution (0.9% NaCl (aq)) was obtained from Baixer Neancate Ashyaa, Oi Schopdae, Znv, and Alina. Cremophor Eb was obtained from §1dta-A1ypsy P1yb, Ca8 (1e NSh, Νδν, AiGaPa. 5 Fluorouracil, a clinical preparation, a clear, colorless liquid was obtained from Maupe Riegta P1yu Bt. , M (\ Woweg1eu, νΚ. ', AiigPa. Penicillin-streptomycin and trypan blue were obtained from §1§ta-Ayg1cb, Saye NSh, Νδν, AikKaya. Hep3B2.1-7 human liver cancer cells were obtained from the American type culture collection ( ATCC), KoskuShe, ΜΌ, I8A. Ma1gsche1® was obtained from VO Vyukšepsek, No. g1k Cuba, Νδν, ASChLaPa.

The use of Ma (m1gе1® in the inoculation suspension improved the tumor uptake rate and reduced tumor size variability, and the growth of human liver cancer Hep3B2,1-7 is more stable when inoculated in the presence of their extracellular matrix.

Preparation and administration of the compound: Cremophor Eb: saline (1: 9, v / v; control excipient), W ^) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6- methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (compound A) or 5-fluorouracil (control compound) was administered according to the scheme below.

Group Compound Dose (mg / kg) Planned treatment Treatment Introduced one Filler The control 10 ml / kg Once a day for 21 days (Day 0 20) Once a day for 19 days (Day 0-18) 2 Compound A 0.2 ml @ 10 ml / kg - 2 mg / kg Once a day for 21 days (Day 0 20) Once a day for 19 days (Day 0-18) 3 Compound A 1.0 ml @ 10 ml / kg = 10 mg / kg Once a day for 21 days (Day 0 20) Once a day for 19 days (Day 0-18) 4 Compound A 5.0 ml @ 10 ml / kg = 50 mg / kg Once a day for 21 days (Day 0 20) Once a day for 19 days (Day 0-18) 5 5-fluorouracil 7.5 ml @ 10 ml / kg = 75 mg / kg Once a week for 21 days (Day 0.7 and 14) Once a week for three weeks (Day 0, 7 and 14) 6 The control 'speed takeover ' Without treatment - -

Control excipient, Cremophor Eb: saline (1: 9, v / v), was administered po in a dosed volume of 10 ml / kg, once a day for 21 consecutive days (day 0-20).

W ^) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, prepared in Cremophor E: saline (1 : 9, v / v). The mother liquor was prepared weekly and stored at 4 ° C. Dose solutions were prepared on each day of administration. W ^) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide in a dosed volume of 10 ml / kg, once a day for 21 days (day 0-20). The compound was administered at doses of 2, 10 and 50 mg / kg.

The clinical preparation 5-fluorouracil was diluted in sterile saline and administered iv. through the tail vein at a concentration of 75 mg / kg, in a dosing volume of 10 ml / kg, once a week for three weeks (on day 0, 7 and 14).

Mice in group 6 were not given any treatment (absorption rate control). On day 7 of the study (21 days after inoculation), the mice were rejected and the liver was opened to determine the rate of absorption and the size of the tumors in the liver wall.

The body weight of each animal was measured immediately before the dose. A volume of 66,020,624 was calculated for administration to each animal and adjusted according to the body weight of the animal.

Tumor measurements. The wet weight of the liver and tumors were measured post-mortem after extraction on the day the study was completed. After the study, the liver was removed from all mice in each study group and weighed. The number of visible tumors, if present, was counted. These tumors were removed from the liver and weighed.

Data Measurement and Sampling Scheme

Measured data Scheme Body weight Day 0, then three times a week (Monday, Wednesday and Friday), and on the day of the end of the study for groups 1 to 5 inclusive. Liver weight and tumor weight The wet weight of the extracted liver and tumor of all mice in groups 1–5 inclusive, posthumously on the day of the end of the study, and from one mouse in group 5, which died on the last day of treatment. Image selection Liver and tumors Of all the mice in group 6 (absorption rate control ’) on day 7 of the study. Liver and tumors Of all the mice in groups 1-5 inclusive, posthumous on the day of the end of the study, and from one mouse in group 5, which died on the last day of treatment. Liver Of all the mice in groups 1-5 inclusive, posthumous on the day of the end of the study, and from one mouse in group 5, which died on the last day of treatment.

Collection and calculation of data. The pulsed transceiver of each animal (Vaeg Sau Ea1a 8u5ΐet5 P1u Во. Vaapu Vau, N8 ^) was scanned using a barcode scanner (LaMax I, Ea1aMag5. 8 \ yP / eG1ag ^) immediately before data collection. All measurements were carried out with constant portable calipers (ЛЬ5о1и1е ОшипаОс Μоάе1 СЭ-6 С8, MPiUuo Sogrogayop, 1arap). The data was synchronized with a secure relational database, UyuPyagt, using ReGadop Rogt5 4.0 (ReGadop® 8Ge Sogrogayop, PieuuShe, 1b, U8A) as data transfer software. AUAM ν2.4 was used for statistical reporting and data calculation.

Statistical processing and calculations. All static processing was performed using 81dta8ΐaΐ 3.0. (8P88 Ai51ga1a51a P1y Εΐά, No. r1H 8yάηeu, N8 ^, A ^ haNa).

Student’s criterion based on a double sample was used to determine the significance of changes in body weight within the treated group from day 0 to the day the study ends. If the data did not meet the normality criterion or the homogeneous variance criterion, then the Mann-Whitney rank sum test was performed.

Univariate analysis of variance (ANOVA) (all pairs of the multiple comparison procedure and multiple comparison relative to the control group) was performed for liver and tumor weight data after the end of the study. If this test did not meet the homogeneous dispersion criterion, then we performed a one-way analysis of variance Kruskal-Wallis (LNOVA) in series. Similar statistical processing was performed for data on mice bearing the tumor in the present study.

p value less than 0.05 was considered reliable.

Liver and tumor weight data for mice bearing the tumor and average liver and tumor weights per mouse per group in the group of mice bearing the tumor.

Group Treatment Average liver weight (g) Stand. mistake Wed The average tumor weight (g) Stand. mistake Wed The number of mice with tumors (out of 10) one Control filler 4,560 0.673 3,382 0.979 4 2 Compound A (E 2 mg / kg 2,775 0.475 1,776 0.576 6 3 Compound A (d> 10 mg / kg 2,551 0.446 1,407 0.465 7 4 Compound A (a) 50 mg / kg 1,677 0.161 0.624 0.257 4 5 5Pi ™ @ 75 mg / kg 1,217 0.051 0.143 0,078 4

No samples were taken from mice from group 5 (5-fluorouracil at 75 mg / kg), which were discarded during the study. Due to the presence of large tumors in some mice, as evidenced by the appearance of a swollen mass in the abdomen, the study was stopped 18 after the start of treatment.

A dose-dependent trend in liver and tumor weight reduction was evident in the groups treated with ^ (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3 67,020,624 dihydroxypropyl) cyclopropane-1-sulfonamide. If we consider only mice bearing a tumor, it was found that the average weight of the liver in the groups treated with ^ (§) - (3,4-difluoro-2- (2 fluoro-4-iodophenylamino) -6-methoxyphenyl) -1 - (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide at the highest dose (group 4 at a dose of 50 mg / kg) and 5-fluorouracil (group 5 at a dose of 75 mg / kg) is significantly different from the group with a control excipient (group 1; p <0.05). It was also found that the average tumor weight in the groups treated with ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (groups 3 and 4 at doses of 10 and 50 mg / kg, respectively) and 5-fluorouracil (group 5 at a dose of 75 mg / kg), significantly differs from the group with a control excipient.

These results are presented graphically in FIG. 16 (average weight of the liver — only mice bearing the tumor) and 17 (weight of the liver tumors — only mice bearing the tumor).

Example 116

Growth responses of orthotopic tumors of the human colon NT-29 in mice No. 1be in the treatment of various amounts of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1 - ( 2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Dose-dependent response with the introduction of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (compound A), regarding inhibition of the development of orthotopic adenocarcinoma of the human colon and rectum, NT-29 was evaluated in BABB / c pi / pi mice, compared with the optimal dose of 5-fluorouracil (75 mg / kg).

Animals. For the study, female BABB / c pi / pi mice were used (itegue OG Abekppbe, Abe Satrik, 8A, Aikbaia), aged 7-12 weeks, with a body weight in the range of 16.58-25.39 g (average value 21, 52 g). The mice were divided into 6 experimental groups (4 treated groups and 2 control groups) as follows: the number of mice per group: 10 in groups 1-5 inclusive; 9 in the control group absorption rate (group 6).

The mice were kept in an environment with controlled conditions (target ranges: temperature 21 ± 3 ° C, humidity 30-70%, air exchange rate 10-15) in isolated conditions (quarantine) with a cycle of 12 hours of light / 12 hours of darkness. Temperature and relative humidity were monitored continuously. The animals were given commercially available food for rodents (Ka! And Moika Sziek, §res1a1bu Reebk P1u Lub, C1ep Rottek !, ^ ek! Etp AikbaAa) and tap water ab NIiit. Food and water were sterilized by autoclaving.

Inoculation of the tumor. HT 29 human colon and rectal adenocarcinoma cells (passage 4 from the working stock UR-§! Osk 325) were cultured in KPM11640 cell culture medium supplemented with 10% PBG and penicillin-streptomycin (final concentration 50 IU / ml). Cells were harvested by trypsinization, washed twice in HB§§ and counted. After that, the cells were resuspended in HB§§ and brought to a final volume containing 2x10 ⁸ cells / ml. Before inoculation, the incision sites were thoroughly washed with alcohol and the incision was made through the abdominal wall to access the cecum. A needle was inserted through the surface of the wall of the cecum, into which 5 μl of cells were injected (1 × 10 ⁶ cells).

Materials The following substances were obtained from appropriate commercial sources.

A sterile saline solution (0.9% ΝηΟ (aq.)) Was obtained from Bax! Et NeaIsate AikbaNa, O1b Toopdae, Ν§ν, AikbaAa. Cremophor Eb was received from §1dta-A1bbsb P! At Yb, Sakbe NSh, Ν§ν, Aikbaia. 5Fluorouracil, a clinical preparation, a clear, colorless liquid was obtained from Maupe Pbacta P1u Lb. KPM11640 cell culture medium, PB§ and HB§§ were obtained from Ipodep. AnkCaPa P! In Yb, M! ^ auebeu, U1C, AikbaIa. Penicillin-streptomycin and Trypanovy blue were obtained from §1§ta-A1bt1cI, Sakbe NI1, Ν§ \ ν, Aik1gaIa. NT 29 human colon and rectal adenocarcinoma cells were obtained from the American Type Culture Collection (ATCC), KoskuShe, MO, and §A.

Preparation and administration of the compound: Cremophor Eb: saline (1: 9, v / v; control excipient), ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 -methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide or 5-fluorouracil (control compound) was introduced according to the scheme below.

- 68,020,624

Group Compound Dose (mg / kg) Planned treatment Treatment Introduced one Filler The control 10 ml / kg Once a day for 21 days (Day 0 20) Once a day for 21 days (Day 0 - 20) 2 Compound A 0.2 ml @ 10 ml / kg = 2 mg / kg Once a day for 21 days (Day 0 20) Once a day for 10 days (Day 0-9) 3 Compound A 1.0 ml @ 10 ml / kg = 10 mg / kg Once a day for 21 days (Day 0 20) Once a day for 21 days (Day 0 - 20) 4 Compound A 5.0 ml @ 10 ml / kg = 50 mg / kg Once a day for 21 days (Day 0 20) Once a day for 8 days (Day 0-7) 5 5-fluorouracil 7.5 ml @ 10 ml / kg = 75 mg / kg Once a week for three weeks (Day 0.7 and 14) Once a week for 3 weeks (Day 0.7 and 14) 6 The control 'speed takeover ' Without treatment -

Control excipient, Cremophor Eb: saline (1: 9, v / v), was administered po in a dosed volume of 10 ml / kg, once a day for 21 consecutive days (day 0-20).

No. (5) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide was prepared in Cremophor E: saline (1 : 9, v / v). The mother liquor was prepared weekly and stored at 4 ° C. Dose solutions were prepared on each day of administration. ^ (5) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide in doses of 2, 10 and 50 mg / kg, in a dosing volume of 10 ml / kg, once a day for 21 days (day 0-20).

The clinical preparation 5-fluorouracil was diluted in sterile saline and administered iv. through the tail vein at a concentration of 75 mg / kg, in a dosing volume of 10 ml / kg, once a week for three weeks (on day 0, 7 and 14).

Mice in group 6 were not given any treatment (absorption rate control). On day 7 of the study (21 days after inoculation), the mice were discarded and the colon was removed to determine the rate of absorption and tumor size in the wall of the cecum.

The body weight of each animal was measured immediately before the dose. The volume for administration to each animal was calculated and adjusted according to the body weight of the animal.

Tumor measurements. The wet weight of the cecum and tumors were measured post-mortem after extraction on the day the study was completed. After the study, the cecum was removed from all mice in each study group and weighed with intact tumors. Then, tumors were removed from the cecum and weighed.

Livers were also removed from all mice in each group after the end of the study and fixed in 10% buffered formalin. Five liver samples from the control-vehicle group were fixed in paraffin, cut and stained with hematoxylin and eosin (H&E) for histological analysis of morphological changes.

Data Measurement and Sampling Scheme

Measurable data Scheme Body weight Day 0, then three times a week (Monday, Wednesday and Friday), and on the day of the end of the study for groups 1-5 inclusive. Cecal Weight and Tumor Weight Excised cecum and tumor from each mouse at the end in groups 1-5 inclusive Image selection Cecum and tumors Of all the mice in group 6 (control поглощения absorption rate ’) posthumously, on day 7 of the study. Cecum and tumors Of all the mice in groups 1-5 inclusive posthumously, on the day of the end of the study, and from the mouse that died during the study period. Liver Of all the mice in groups 1-5 inclusive posthumously, on the day of the end of the study, and from a mouse that died in the period research.

Collection and calculation of data. The pulse transceiver of each animal (Wag Sob Ea1a 5uz1ets

- 69 020624

P1y Πά, Boiapu Wau, \ 3H) was scanned using a barcode scanner (LaMax I, BaHMagz, 3 \\ h1 / e1a0) immediately before data collection. All measurements were carried out with constant portable calipers (Alcoe Bx ^ alcois Mo0e1 SE-6 C3, Miuooo SogrogNop, 1arap). The data was synchronized with the secure relational database U1uoPbag, using Repygadop Rogsz 4.0 (Repygadop® ZoPmage SogrogNop, PieGuuShe, 1b, I3A) as data transfer software. AP) AM ν2,4 was used for statistical reports and data calculation.

Statistical processing and calculations. All static processing was carried out with the help of 3 §§31a! 3.0. (3P33 Ai81ga1a81a P1y Πά, ShgSh 3y0peu, Ν3 \ ν, Ai81gaNa).

Student’s criterion based on a double sample was used to determine the significance of changes in body weight within the treated group from day 0 to the day the study ends. In the groups treated with No. (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide in doses of 2 and 50 mg / kg, treatment was discontinued earlier due to excessive loss of body weight. In these groups, a student-based test based on a double sample was used to determine the significance of changes in body weight within the treated group in the interval from day 0 to the final day of treatment in the experiment, and in the interval between the last day of treatment and the day the experiment ended. If the data did not meet the normality criterion or the homogeneous dispersion criterion, then the Mann-Whitney rank sum test was performed.

Univariate analysis of variance (ΑNOVΑ) (all pairs of the multiple comparison procedure and multiple comparison relative to the control group) was performed for data on the weight of the cecum and tumor weight after the end of the study. If the data did not meet the normality criterion, then the values were turned into the natural logarithm before the procedure.

A p value of less than 0.05 was considered reliable.

Observations In all the studied groups, the average loss of body weight was measured, including the control group with filler. Diarrhea and signs of dehydration (loss of skin elasticity) were observed in all studied groups, including the control excipient. Severe body weight loss in the early stages of the study led to discontinuation of treatment in groups treated with Ν- (3) - (3,4difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3 -dihydroxypropyl) cyclopropane-1sulfonamide at the lowest dose (2 mg / kg) and the highest dose (50 mg / kg) on day 9 and day 7 of the study, respectively. Since body weight loss was less severe in the group receiving N- (3) (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane -1 sulfonamide at a dose of 10 mg / kg, then all treatments for this group were administered according to the scheme. The average body weight loss after the end of the study for this group and the group treated with 5-fluorouracil was significant.

Despite the fact that the rate of absorption of NT-29 tumors in the group, the rate of absorption 21 days after inoculation was 100%, the size of these tumors was much smaller than expected. This may be due to the lack of significant differences in the average weight of the cecum and the weight of tumors between the groups treated with No. (3) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- ( 2,3-dihydroxypropyl) cyclopropane-1-sulfonamide - and a control group with a filler. Also, no effect of 5-fluorouracil on the weight of the cecum and NT-29 tumor was observed.

Body weight measurements (± standard error, cf.) (end days of treatment and end date of the experiment)

Gr. Compound Dose (mg / kg), route, schedule Host response Body weight difference (g) (± Stand. Error cf.) End day of treatment % difference in body weight End day of treatment Body weight difference (g) (± STD error cf.) End day of experience % difference in body weight End day of experience number Survivors (Number of living / Total) one Control filler - by. Once a day for 21 days (Day 0 - 20) - - -0.8 ± 0.4 -3.6 8/10 2 Compound A 2 by. Once a day for ten days (Day 0-9) -2.4 ± 1.4 (Day 9) -11.1 0.1 ± 0.9 0.4 4/10 3 Compound A 10 by. Once a day for 21 days (Day 0 - 20) - - -1.5 ± 0.3 -6.9 7/10 4 Compound A fifty by. Once a day for eight days (Day 0-7) -3.8 ± 0.5 (Day 7) -17.8 -1.3 ± 0.9 -5.9 7/10 5 5-fluorouracil 75 V.V. Once a week for three weeks (Day 0, 7 and 14) - - -3.3 ± 0.4 -15.2 8/10

Body weight data were not collected for group 6 (absorption rate control). The group was rejected on day 7 of the study (day 21 after inoculation) to visually assess the growth of adequate growth

- 70,020,624 tumors for the purposes of this study.

Treatment was discontinued in the 2 ^ - (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) 1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide group at a dose 2 mg / kg) on day 9 of the study and in the 4 ^ - (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) group 50 mg / kg cyclopropane-1-sulfonamide) on day 7 of the study, since mice experienced very intense weight loss. All remaining groups received all of the planned treatments during the trial period.

The average tumor weight in each group is shown in FIG. 18. The average tumor weight for each group includes only those animals that remained alive on the final day of the experiment. Values for mice that died during the study period were not included in the calculated mean values.

Cecal weight and tumor weight data

1 ruppa Treatment ΙΓ) animal Weight blind guts (AND Her tumors (!) Middle dog blind guts (one) Vultures. mistake Wed Middle weight opumish (AND Stand. mistake er one Control ίί filler ί Krsmofor R. 1 /(.dik'vpn solution) 173811 170774 17072 ‘» 173673 171429 175732 171539 173576 1 7 (1836 172014 0.290 0.331 0.267 0,307 0.31 1 0.946 0.286 0.287 0.397 0.506 0.070 0.160 0.017 0.005 0.326 0.627 0.058 0.002 0.000 0.176 0.413 0.08 1 0.149 0,080 2 Compound Λ ί / ζ ^Ί 2 MR'KG 175867 170936 172003 176472 171358 170825 174466 171587 0.347 0.170 0.150 0.205 0.377 0.235 0.251 0.322 0.001 0.001 0,000 0.005 0.0 (11 0.122 (1.005 0.003 0.296 0.033 0.053 0,030 s Compound Α 10 ΜΙ 'ΚΙ 173623 0387 0,000 0.244 0,025 0.078 0.059 170793 173304 175676 0.198 0.258 0,1 1 1 0,000 0.257 0.008

- 71 020624

[ruppa Treatment Yu zhntppogo Her blind guts (AND Her tumors (AND Middle carried blind guts (and Stand. mistake er Middle her tumors (D) Sta sq. mistake er 1 7! (105 1 7! 466 176586 170858 175862 171364 0.263 0.237 0.234 0.289 0.320 0.251 0.422 0.001 0.014 0.004 0.100 0,000 4 Compound Λ 50 mg / κι 175697 171349 1-74272 176355 171041 174536 175656 ^_ 175626 171457 174501 0) 230 Κ254 0.238 0.201 0.337 0.169 0.528 0.217 0.5 1 2 0.463 0.002 0.002 0,000 0.004 0.166 0.655 0.001 0.001 0.001 0.026 0.290 0.038 0.122 0,092 5 PGI ¹¹¹ and diva 75 μγ / κι 171322 175559 0) 355 0.199 0.245 0.001 0.391 0,0.40 0.069 0.04 1 176502 0.360 0,000 176241 [75857 176242 174165 176417 170592 0.284 0.421 0.706 0.415 0.527 0.237 0.377 0.000 0.010 0.329 0.130 0.079 0.000 171501 0.005

Shaded icons indicate samples collected from mice that died during the study period. From the calculated average values for the weight of the cecum and tumor weight, these values are excluded. The trend indicates a decrease in NT-29 tumors and cecal weight data after treatment with 10 mg / kg L- (8) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide.

Example 117

Tumor growth inhibition in nude mice bearing A375 human melanoma xenografts.

Six groups (n = 9) of mice with tumors were used. The control groups included: one group, which received 10% cremophor Eb / saline filler by forced oral nutrition (po), once a day for 14 days (cbx 14), and the second group, which received paclitaxel as a comparative agent a dose of 30 mg / kg by injection through the tail vein (s), every other day, a total of five doses (s.) obh5). Four test groups received orally L- (8) - (3,4-difluoro-2- (2-fluoro4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (compound A ) at a dose of 25 mg / kg or 50 mg / kg, cbh 14, or at a dose of 12.5 or 25 mg / kg, Yb x 14. The treatment outcome was evaluated using Τ0Ό, defined as the difference in the average time to the control point of tumor volume in treated group compared with the control group. Toxicity was evaluated by measuring body weight and clinical observations.

Animals. The experiments were performed on female atimic nude mice (pi / pi, Nag1ap) aged 10 to 11 weeks and body weight (B ^ U) in the range from 19.3 to 25.5 g on day 1 of the study. The animals were provided with access to water (reverse osmosis, 1 ppm С1) and feed No. H 31 Mobb apb! Gab1a! Eb baB O1e1®, consisting of 18.0% crude protein, 5.0% crude fat and 5 0% crude fiber. The mice were kept in irradiated cages for laboratory animals ABPNA-Όή® ебeb-o'COЫ® in stationary micro-isolators with a 12-hour light cycle at a temperature of 21-22 ° C (70-72 ° P) and a humidity of 4060%. The recommendations of Oshbe ог С sage apb ike £ а а ога ога Ю Ат т Attack were respected regarding conditions, care, surgical procedures, nutrition and water supply, and veterinary care. Tumor implantation. Xenografts were obtained from A375 human melanoma tumors by serial dilutions of atimic nude mice. A A375 tumor fragment (~ 1 mm ³ ) was implanted subcutaneously in the right flank of each test mouse and tumor growth was observed until an average volume of 100-72 020 624 was reached.

150 mm ³ . After thirteen days, designated as day 1 of the study, the animals were divided into six groups, each of which included nine mice (reduced from ten) with individual tumor volumes in the range of 63 to 221 mm ³ and a group average tumor volume in the range of 125.3 up to 125.9 mm ³ . Tumor volume was calculated according to the formula

where ν is the width and

- the length in mm of the A375 tumor.

Materials No. (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide was dissolved at a concentration of 5 mg / ml in 10 % cremophor Eb in saline when sonicated, stirred and heated to 35 ° C to facilitate dissolution. A solution of 5 mg / ml served as a dosage solution for treatment at a concentration of 50 mg / kg, and dosage solutions for treatment at concentrations of 25 and 12.5 mg / kg were obtained by serial dilution. Dosage solutions were stored up to one week at room temperature in a dark place.

Dosage solutions of paclitaxel (ΝΈΙ) were prepared from 30 mg / ml stock solution for daily use by diluting to 3 mg / ml in 5% ethanol, 5% cremophor E in 5% dextrose in water (I5). Paclitaxel was dosed at 30 mg / kg.

Treatment. The table below shows the treatment regimen.

Treatment regimen Group η Means mg / kg way Scheme one nine Filler by. ςά x 14 2 nine Paclitaxel thirty V.V. ηοά x 14 3 nine Compound A fifty by. x 14 4 nine Compound A 25 by. B x 14 first day 1 dose 5 nine Compound A 25 by. ςά x 14 6 nine Compound A 12.5 by. B x 14 first day 1 dose

The mice in group 1 received an excipient consisting of 10% Cremophor E in saline by forced oral (daily) oral administration in fourteen doses (c | bx14), and they were control for analysis of tumor progression. The animals in group 2 were injected with intravenous (s) paclitaxel as a comparative agent at a concentration of 30 mg / kg, every other day in the form of five doses (cobx5). Mice in groups 3-6 received orally ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide according to the following appropriate regimens: 50 mg / kg, ςάχ 14; 25 mg / kg, twice a day for 14 days with a single dose on the first and last days (Ybh14); 25 mg / kg, ςάχ 14; 12.5 mg / kg, Lbx 14. All doses were administered in a volume of 0.2 ml per 20 g of body weight, and brought into line with the body weight of the animal.

End point. Tumors in all groups were measured twice a week using calipers. Each animal was killed after reaching a tumor of the size of the endpoint of 2000 mm ³ or on the final day of the study (day 60), depending on which of these events occurs first. The time to the control point (TTE) for each mouse was calculated according to the following equation:

. 1st „, (end point volume, mm ¹ ) - b

TTE (days) ^_ '

Where G is the line segment and m is the slope of the line obtained by linear regression of the 1 transformed tumor growth dataset.

The data set consisted of the first observation that exceeded the volume of the control point of the study and three consecutive observations that immediately preceded the achievement of the volume of the control point. Animals that did not reach the control point were assigned a TTE value equal to the value on the last day of the study. Animals classified as No. HK (unrelated to treatment) death due to an accident (No. HKa) or due to unknown causes (No. HKi) were excluded from the TTE calculations (and all further analyzes). Animals classified as TK (treatment-related) deaths or No. HKT (unrelated to treatment, due to metastasis) were assigned a TTE value equal to that on the day of death.

The result of treatment was determined from tumor growth retardation (TOY), defined as an increase in the average time to the control point (TTE) in the treated group compared to the control group: TOI = T-C, expressed in days, or as a percentage of the average TTE of the control group :

- 73 020624% mc-— £ ^χ 100, s

where T is the average value of the TTE for the treated group;

C is the average value of the TTE for the control group (group 1).

Treatment may cause partial regression (SC) or complete regression (SC) of the tumor in the animal. In the RK response, the tumor volume is 50% or less of its volume on day 1 for three consecutive measurements during the study and is equal to or greater than 13.5 mm ³ for one or more of these three measurements. In the SC response, the tumor volume is less than 13.5 mm ³ for three consecutive measurements during the study. An animal with a SC response at the end of the study was further classified as tumor-free survivor (TP8). Tumor regression was monitored and recorded.

Side effects. Animals were weighed daily for the first five days of the study and then twice a week. Animals were often monitored to detect visible signs of any unwanted treatment-related side effects, and clinical symptoms were recorded when detected. Permissible tolerance is defined as the average group weight loss of less than 20% during the experiment and not more than one death associated with treatment in the group of animals. Any dose regimen that does not meet this criterion is considered to exceed the maximum tolerated dose (MTE). Death is classified as TK if it can be caused by side effects of treatment, as evidenced by clinical symptoms and / or autopsy, or can be classified as TK if the cause is unknown during the dosing period or within 10 days from the last dose. Death is classified as ETC if there is no evidence that death is associated with side effects of treatment.

Statistical and graphical analyzes. The log-rank criterion was used to analyze the significance of differences between the TTE values of the treated and control groups. Bilateral statistical analyzes were performed at a confidence level of P = 0.05.

Curves of mean tumor growth values are plotted versus time. If the animal was excluded from the study due to tumor size or TK death, then the final tumor volume recorded for the animal was included in the data used to calculate the group mean value of the tumor volume at subsequent times. Curves were truncated after 50% of the animals in the group were excluded from the study due to tumor progression. Kaplan-Meyer plots were built to demonstrate the percentage of animals remaining in the study, depending on time, and used the same data set as for the log-rank criterion. We used Rgbt (SarbPab) for \ νίηbo \\ ъ 3.03 for all graphical presentations and statistical analyzes.

The generalized results of the treatment effect

The growth of A375 tumors in control mice (group 1). Animals in group 1 received a 10% cremophor Eb / saline filler, in dB / 14. Tumors in control mice grew progressively up to 2000 mm in the volume of the control point with an average CT of 22.8 days, setting the maximum possible TS in the study of 37.1 days, or 163% TSE.

The effect of treatment in paclitaxel (group 2). The animals in group 2 were injected with paclitaxel as a comparative agent, 30 mg / kg, iv, ext / 5. All nine animals reached the tumor volume at the endpoint. Tumors grew in parallel and were slightly shifted to the right compared to the control group. The average TTE value was 28.8 days, which corresponds to 26% TSE, a reliable result according to the log-analysis (Table 2, P = 0.0088 C1 rel. C2). No tumor regression associated with paclitaxel treatment was observed.

Effect of treatment ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide (groups 3-6): Groups 3-6 received an oral dose of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide as monotherapy. Animals in group 3 were injected with 50 mg / kg according to the DB / 14 regimen. Nine tumors in the group reached volume at the endpoint of the study.

- 74,020,624

The mean tumor volume in the group underwent a slight net change during the first ~ 10 days, then increased during the study. In one animal, PK of the tumor was observed. The average TTE value was 27.5 days, or 21% LLP, a reliable result (P = 0.0054 01 νδ. O).

Animals in group 4 received 25 mg / kg ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide according to the scheme Ybh14. Four of the nine animals in the group remained on day 60, all TP8. Additionally, 2 out of 9 animals had tumors that reached the endpoint volume of the study one day before the end of the study. The group had 4/9 RK, 5/9 SK and 4/9 TP8. The average tumor volume has been decreasing since the first few days of the study and lasted for approximately 30 days. Re-grown tumors in 5 of 9 animals were counted to restore the average tumor growth, starting from approximately day 32 and continuing until the end of the study. The average group TTE of the value was 59.9 days, characterizing the maximum possible 163% of the LLP (P <0.0001, Table A1).

Mice in group 5 also received 25 mg / kg ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1- sulfonamide, but according to a less intensive regimen with | bx 14. All nine animals in group 5 reached the endpoint tumor volume, without tumor regression. Tumor growth was carried out almost in the same way as in the control group. The average TTE was 25.6 days, or 12% LLP, an unreliable result (P = 0.0662 O1 νδ. O5).

The animals in group 6 were administered 12.5 mg / kg ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1 -sulfonamide according to the scheme Ybh14. All tumors in the group reached volume at the endpoint of the study. As in group 4, the average tumor volume in group 6 decreased at the beginning of the study, but this decrease persisted only for approximately nine days and was associated with a single PK response. Tumor volume increased from day 10 until the end of the study. The average TTE for the group was 27.5 days, which corresponds to a reliable 21% LLP (P = 0.0424 O1 rel. O6).

In conclusion, No. (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide exhibits dose-dependent antitumor activity in relation to xenografts of A375 human melanoma with oral administration of doses, both once a day and twice a day. Dosing twice a day has an improved effect compared to a single dose on the amount of received LLP and on the number of objective responses. Thus, the antitumor activity of ^ (§) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6 methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide depends on the dose , and the introduction scheme.

Example 118

Activity against subcutaneous xenografts of Co1205 human colon cancer.

Animals. We used female atimic nude mice (II / II, Nag1ap) aged 12 to 13 weeks and body weight (Βν) in the range from 18.3 to 27.3 g on day 1 of the study. The animals were given access to abPI to water (reverse osmosis, 1 ppm С1) and feed No. H 31 MobSheb apb 1gab1a1sb bab Όίc1®, consisting of 18.0% crude protein, 5.0% crude fat and 5.0% crude fiber. The mice were kept in irradiated cages for laboratory animals ALPNA-Op® Leb-o'CO3 8® in stationary micro-isolators with a 12-hour light cycle at a temperature of 21-22 ° C (70-72 ° P) and a humidity of 40-60%. The recommendations of Oshbe Gog Sage ap Ive OG LábaFu ApipaR regarding conditions, care, surgical procedures, nutrition and water supply, and veterinary care were followed.

Tumor implantation. Xenografts were obtained from Co1205 cells of human colon cancer. Tumor cells were cultured in 10% heat-inactivated fetal bovine serum, 100 units / ml sodium penicillin O, 100 μg / ml streptomycin sulfate, 0.25 μg / ml amphotericin B, 25 μg / ml gentamicin, 2 mM glutamine, 1 mM pyruvate sodium, 10 mM HEPE8 and 0.075% sodium bicarbonate. Cell cultures were maintained in tissue culture flasks in a humid chamber at 37 ° C, in an atmosphere of 5% CO ₂ and 95% air. On the day of tumor cell implantation, Co1O 205 cells were harvested by logarithmic growth and resuspended in a 50% Ma1pde1 matrix (ΒΌ VuvSK epsev) in ΡΒ§ at a concentration of 5 × 10 ⁶ cells / ml. Each experimental mouse received 1 x 10 ⁶ CoIo 205 cells implanted subcutaneously in the right flank, and tumor growth was observed until an average size of 80-120 mm ³ was reached. Fourteen days later, designated as day 1 of the study, the animals were divided into eight groups (n = 9) with individual tumor volumes in the range of 63 to 196 mm ³ and group average tumor volume in the range of 118-119 mm ³ . Tumor size was calculated according to the formula

where ν is the width and

- length in mm of Co1o205 tumor.

Tumor weight can be estimated assuming that 1 mg is equivalent to 1 mm ³ of tumor volume.

- 75,020,624

Materials Dosage solutions of compound A were prepared fresh daily by dissolving the required amount of the compound in 100% cremophor Eb, and then diluted ten times in physiological saline. The final dosed concentrations in the solution were 2.5, 5, 10 or 20 mg / ml, to provide the appropriate dose of 25, 50, 100 or 200 mg / kg in a dosing volume of 10 ml / kg. Paclitaxel (No. 1igl Pactaseis 1cp) was prepared fresh every day by dosing in an excipient consisting of 5% ethanol and 5% cremophor EL in 90% B5 / V (5% EC excipient).

Treatment.

The table below shows the treatment regimen.

Treatment regimen Group η Means mg / kg Way Scheme one nine Filler - BY. c] b x 14 2 nine Paclitaxel thirty V.V. Yaob x 14 3 nine Compound A fifty by. Yab x 14 4 nine Compound A 25 by. Yb x 14 first day 1 dose 5 nine Compound A 25 by. apple x 14 6 nine Compound A 12.5 by. Yb x 14 first day 1 dose

Group 1 received an excipient preparation (10% Cremophor E in saline), and it was a control group for growth. Group 2 received the comparative drug paclitaxel, which was administered in its optimal regimen to nude mice (30 mg / kg iv with | v / 5). Groups 3-6 received doses of 25, 50, 100 and 200 mg / kg, respectively, of compound A, which were administered po c | b / 14, while dosing in group 6 (200 mg / kg) was canceled after six days due to toxicity. All doses were aligned with the body weight of the animal (0.2 ml per 20 g of body weight).

End point. Tumors were measured twice each week using calipers. Each animal was killed after the tumor reached a predetermined endpoint size of 2000 mm ³ or on the final day of the study (day 74), depending on which of these events occurs first. However, the control tumors did not exhibit logarithmic growth characteristics after reaching a size of about 800 mm ³ . Therefore, a tumor size of a control point of 800 mm ^{3 was} used to analyze tumor growth retardation (T0B). The time to the control point (TTE) for each mouse was calculated according to the following equation:

, 1st _{1 (1} (end point volume, mm ² ) -3

TTE (days) = ......-...........——-, t

where b is a line segment and m is a line slope obtained by linear regression of a 1 transformed tumor growth dataset.

The data set consisted of the first observation that exceeded the volume of the control point and three consecutive observations that immediately preceded the achievement of the volume of the control point. Animals that did not reach the control point were assigned a TTE value equal to the value on the last day of the study. Animals classified as ΝΚΚ (unrelated to treatment) death due to an accident (No. HKa) or due to unknown causes (No. HKi) were excluded from the TTE calculations (and all further analyzes). Animals classified as TC (treatment-related) deaths or GCT (non-treatment-related deaths due to metastasis) were assigned a TTE value equal to that on the day of death.

The result of treatment was determined from the delayed tumor growth (T0B), defined as an increase in the average time to the control point (TTE) in the treated group compared to the control group: T0B = T - C, expressed in days, or as a percentage of the average TTE of the control group :

% TCO = ^G_C x 100,

C where T = average TTE for the treated group;

C = mean TTE for the control group.

The control group was defined as group 1 mice.

Treatment may cause partial regression (SC) or complete regression (SC) of the tumor in the animal. In the PK response, the tumor volume is 50% or less of its volume on day 1 for three consecutive measurements during the study, and is equal to or greater than 13.5 mm ³ for one or more of these three measurements. With an SC response, the tumor volume is less than 13.5 mm ³ for three consecutive measurements during the study. Tumor regression was monitored and recorded.

Side effects. Animals were weighed daily for the first five days of the study and then twice a week. Mice were often monitored to detect visible signs of any unwanted treatment-related side effects, and clinical symptoms were recorded when detected. Acceptable toxicity is defined as the average group weight loss of less than 20% when

- 76 020624 experience and no more than one death associated with treatment (TC) among ten treated animals, and any dose regimens that result in higher toxicity are considered to exceed the maximum tolerated dose (MTO). Death is classified as TK if it can be caused by side effects of the treatment, which is confirmed by clinical symptoms and / or autopsy, or can be classified as TK if the cause is unknown during the dosing period or within 10 days after the last dose. Death is classified as ΝΕΡ if there is no evidence that death is associated with side effects of treatment. Animals were often monitored to detect side effects and VU measurements. VU changes were unremarkable, and all treatments were tolerably tolerable, with the exception of the group

6. Dose six times a day, po 200 mg / kg of Compound A resulted in one death, estimated at day 7, and two additional TK deaths on day 8. All mice in group 6 had clinical toxicity symptoms, including twisted positions, hypoactivity, and loose stools.

Statistical and graphical analysis. The log-rank criterion was used to analyze the significance of differences between the TTE values of the treated and control groups. Bilateral statistical analyzes were performed at a confidence level of P = 0.05.

Curves of mean values of tumor growth represent group average values of tumor volume, graphically presented on the 1st scale as a function of time. If the animal was excluded from the study due to tumor size or TK death, then the final tumor volume recorded for the animal was included in the data used to calculate the group mean value of the tumor volume at subsequent times. Curves were truncated after 50% of the animals in the group were excluded from the study due to tumor progression or after the second TC death in the group. Kaplan-Meyer plots were built to demonstrate the percentage of animals remaining in the study, depending on time, and used the same data set as for the log-rank criterion. We used Rpkt (OgarRaf for Uto 3.03 for all graphical presentations and statistical analyzes.

The generalized results of the treatment effect

Group Average Tc % ΤΟϋ 88 MTU (P) Day 74 number Average VHU RK SC TP8 TC ΝΤΚ one 41.0 - - - 322 (2) 0 0 0 0 0 - 2 60.0 19.0 46% n.d. 0 (3) one 0 0 2 2 - 3 47.9 6.9 17% N.d. 0 (3) one 0 0 2 2 - 4 59.1 18.1 44% N.D. 195 (ABOUT 4 0 0 0 0 - 5 74.0 33.0 80% N.D. 320 (5) 4 0 0 one 0 - 6 57.8 16.8 41% Well. 0 (3) one 3 0 2 2 0.1% Day 22

The growth of Co1205 tumors in control mice (group 1).

In group 1, tumors showed slow, heterogeneous growth. Tumors 7 of 9 treated with vehicle of group 1 control mice reached a tumor volume of the endpoint of 800 mm ³ and two mice remained at the end of the study. In group 1, the average TTE was 41.0 days, and thus the maximum LLP possible in this 74-day study was 33.0 days (80%).

The effect of treatment with paclitaxel (group 2).

Eight mice in group 2 (n = 9) who received paclitaxel treatment remained in the study on day 74 with MTU 143 mm ³ . This corresponds to the maximum possible LLP (33.0 days or 80%) and statistically significant activity (P = 0.002). Five RK responses are documented. The average tumor growth curve showed a decrease in MTU until day 19, with subsequent minor changes until day 47, when tumor growth resumed.

Effects of treatment using compound A (groups 3-6).

In groups 3, 4, 5, the average values of TTEc were observed 47.9, 59.1 and 74.0 days, respectively. For groups 3 and 4, unreliable log-rank results were obtained, and in group 5, the log-rank criterion reached borderline reliability (P = 0.058). These treatments produce a dose-dependent number of regressions, however, the type of regressive response (RK rel. SC) and the number of survivors on day 74 per group does not correlate with the dose. Curves of mean values of tumor growth exhibit similar activities for three dose levels at the beginning of the study (up to day 29), with subsequent dose-dependent delays in tumor re-growth. Three TC deaths occurred in group 6, and the dose was stopped after day 6. Therefore, treatment at a dose of 200 mg / kg was considered to be higher than MTO and measurable for LLP.

Compound A exhibits a dose-dependent activity with respect to xenografts of Colo205 colon cancer. When administered at a dose of 25 mg / kg, compound A exhibits 3% LLP. At a dose of 50 mg / kg, compound A exhibits 46% LLP. Treatment at a dose of 100 mg / kg is characterized by acceptable tolerance, and, similarly to paclitaxel treatment, leads to the maximum possible ΤΟΌ in the experiment with a similar number of regressive responses. Treatment at a dose of 200 mg / kg causes 3 deaths of TA from 9 animals and exceeds MTE. There is a more noticeable initial decrease in tumor mass for compound A compared to paclitaxel; however, the duration of this action was shorter. Re-growth of tumors for groups with doses of 25 and 50 mg / kg initially occurs at a faster pace compared with the control, and at the end of the study, MTU reaches such values for control. Treatment at a dose of 100 mg / kg does not show such rapid re-growth, it exhibits faster tumor growth compared to growth with paclitaxel treatment.

Example 119

Clinical studies in humans.

A randomized, double-blind, open, historical control, safety / efficacy clinical trial in phase I was performed in humans using Compound A relative to placebo in patients who had not previously received chemotherapy with locally advanced or metastatic pancreatic cancer. .

The primary objective of the study was to evaluate the safety and tolerability of compound A. The secondary objective of the study was to evaluate the response rate, clinical benefit, and tumor shrinkage after treatment with compound A. Additionally, the study was planned in such a way as to evaluate the time to disease progression and overall patient survival with pancreatic cancer. Additionally, the pharmacodynamic changes in the parameters of the blood supply to the tumor (including, for example, blood flow, blood volume, time to peak curve of the characteristics of the CBS-recipient operator) were evaluated using ESE-MK1.

In addition, biological markers such as MEK1 and MEK2 genetic polymorphisms and serum proteomics were used to compare the results. You can also use the determination of the degree of accessibility of tumors for resection, as well as evaluate the MTE for compound A.

In the study, Compound A was administered in various doses of approximately 1 mg, approximately 1.5 mg, approximately 2 mg, approximately 2.5 mg, approximately 3 mg, approximately 3.5 mg, approximately 4.0 mg, approximately 4.5 mg, approximately 5 mg, approximately 5.5 mg, approximately 6 mg, approximately 6.5 mg, approximately 7 mg, approximately 7.5 mg, approximately 8 mg, approximately 8.5 mg, approximately 9 mg, approximately 9.5 mg, approximately 10 mg, approximately 10.5 mg, approximately 11 mg, approximately 11.5 mg, approximately 12 mg, approximately 12.5 mg, approximately 13 mg, approximately 13.5 mg, approximately 14 mg, approximately 14.5 mg, or approximately 15 mg.

Criteria for inclusion in the study were based on the following factors.

Histologically / pathologically locally advanced unresectable or borderline unresectable pancreatic cancer, and without signs of metastatic disease.

The diagnosis of locally advanced unresectable pancreatic cancer was established on the basis of a two-phase CT scan and / or endoscopic ultrasound (EI8) (EI8 is described in Appendix P).

A measurable disease according to KES18T and obtained using a two-phase CT scan for 14 days before registration for the study protocol.

Tumor size is greater than or equal to 2 when examined using a two-phase CT scan.

The sufficient function of the organ is documented within 14 days of registration, which is confirmed by: the absolute number of neutrophils> 1500 / mm ³ ; platelet count; 100 thousand / mm ³ ; hemoglobin ³ 9 g / dl in the absence of the need for a blood transfusion in the previous 4 weeks; total bilirubin <1.5 times higher than the upper limit of normal (ϋΡΝ); transaminases (A8T and / or ABT) <2.5χυΡΝ; RT (or ΙΝΚ) <1.5χυΡΝ and ARTT within normal limits (patients who received anticoagulant treatment with drugs such as warfarin or heparin may be included in the study; for patients receiving warfarin, parameters should be carefully monitored at least weekly until ΙΝΚ becomes stable according to measurements with preliminary administration, which is determined using the local standard of care; creatinine clearance> 60 ml / min, calculated according to the formula SosksgoG1-Sai11.

Criteria for exclusion from the study were: previous treatment with compound A within 6 months before registration; a clinical sign of invasion of the tumor into the mucous membrane of the duodenum (which is documented according to endoscopy or endoscopic ultrasound examination); outpatient surgery (e.g., aspiration diagnostic puncture or aspiration biopsy) within 14 days of registration of the study; extensive surgery, significant traumatic injury, or a severe non-healing wound, ulcer, or bone fracture within 21 days of study registration; any of the following symptoms within 6 months before the drug administration study: severe / unstable angina (symptoms of angina pectoris at rest), re-occurring angina (starting within the last 3 months) or myocardial infarction, congestive heart failure, 78 020624 ventricular arrhythmia requiring antiarrhythmic treatment; a history of thrombosis or embolism, such as acute cerebrovascular accident or transient cerebrovascular accident within the past 6 months; history of aneurysm or arteriovenous congenital malformation; the presence of infection with human immunodeficiency virus (HIV) or chronic hepatitis B or C; active clinically severe infection in excess of degree 2 according to CTCA 2; taking any research tools within 4 weeks of registration of the study; uncontrolled hypertension, defined as systolic blood pressure above 150 mmHg. or diastolic pressure above 90 mmHg, despite optimal medical treatment; pulmonary hemorrhage / episodic bleeding, greater than grade 2 according to CTCAE 2 within 4 weeks of study registration; any other bleeding / episodic bleeding greater than grade 2 according to CTCAE 3 within 4 weeks of study registration; case or history of the disease hemorrhagic diathesis or coagulopathy, daily treatment with aspirin or other non-steroidal anti-inflammatory drugs; use of hypericum, rifampin (rifampicin), ketoconazole, itraconazole, ritonavir, or grapefruit juice; known or suspected allergy to compound A; any condition that impairs the patient’s ability to swallow whole pills; any suction problems; another serious, acute or chronic medical or psychiatric condition, or deviation of laboratory parameters from the norm, which may increase the risk associated with participation in the study or study of drug administration, or may interfere with the interpretation of the results of the study, and, in the opinion of the researcher, may be unsuitable for patient inclusion in this study; a history of collagen disease; any contraindications for the implementation of magnetic resonance imaging.

Example 120

Clinical study in humans.

A randomized, double-blind, open, with historical control, distribution in separate groups, safety / efficacy clinical trial in phase I in humans using compound A in patients who had not previously undergone chemotherapy, locally advanced or metastatic gastric cancer was performed as described in the example 117, except that the patients included in the study were diagnosed with either lymphoma, gastric stromal tumors, or carcinoid tumors of the stomach.

Example 121

Carrageenan-induced paw edema (CPE) in rats.

Compound A (6, 20, and 60 mg / kg) or indomethacin (3 mg / kg) was administered orally 2 hours before injection of a 1% suspension of carrageenan into the right hind paw of male 8gadie-OaMeu rats (Ν = 6 per treatment group). Hind paw edema was measured after 3 hours by measuring paw volume using plethysmography. A decrease in hind paw edema of 30% or more indicated significant anti-inflammatory activity. Indomethacin (1pyo) was used as a positive control drug. In FIG. 22 presents data on the increase in paw volume in each of the treated groups, indicating that the oral administration of compound A leads to significant anti-inflammatory activity in the rat paw edema model caused by carrageenan for all doses in the experimental groups.

Example 122

A study of inflammatory arthritis caused by adjuvant in rats.

In an adjuvant-induced arthritis model in rats, Freund's complete adjuvant (CPA) was injected into the right hind paw of rats to induce a pathology similar to rheumatoid arthritis in humans. Compound A was administered orally for 5 consecutive days at doses of 2, 6 and 20 mg / kg. Dexamethasone in doses of 5 mg / kg was also administered orally for 5 days. Enbrel at a dose of 10 mg / kg was administered by subcutaneous injection on days 1 and 4. CPA was injected into the right hind paw one hour after the first dose was administered on day 1. The percent inhibition of swelling of the right hind paw was measured relative to control animals that were injected with days 1 and 5 for the acute phase, while for the delayed phase, the percent inhibition of swelling of the left hind paw was determined relative to the control animals that were injected with filler, on days 14 and 18. Polyarthritis was evaluated in the presence of swelling of the anterior la , Tail, nose, or ears.

In FIG. 23A and 23B show the percent inhibition of swelling relative to control for different treatment groups. Compound A at a dose of 20 mg / kg shows a significant decrease in swelling in both acute and delayed phases. To assess polyarthritis, all 6 animals in the treated group that were injected with a bulking showed swelling of the front legs and tail. For the group with a dose of 20 mg / kg of compound A, 2 of 6 animals did not show swelling of the forepaws and 4 of 6 animals did not show swelling of the tail. For the enbrel-treated group, there was no protection for the animals regarding swelling of the forepaws and no tail swelling was found in 3 out of 6 animals.

Example 123

Inhibition of Arthritis Induced by Collagen Antibody (CA1A) in Mice.

- 79,020,624

Male Ba1b / s mice (Ν = 8 per treatment group) were injected intravenously (tail vein) with 2 mg cocktail of anti-collagen antibody (Cloibgeh) on day 0. KEEA119 (1, 3 and 10 mg / kg RE) or dexamethasone (1 mg / kg RE) was administered for 0-4 days, while enbrel was administered subcutaneously on days 1 and 3. An intraperitoneal injection of Lp§ (50 mg) was administered on day 3 to all mice, with the exception of animals not subject to experiment. The degree of arthritic manifestations was assessed on all limbs and the data were presented in FIG. 24 (maximum rating 16). For all tested drugs and comparative drugs, the presence of significant anti-inflammatory activity was noted. Enbrel and dexamethasone were used as positive controls.

Example 124

The study of cell proliferation ίη νί \ Ό.

A method for determining cell proliferation, including counting cancer cells treated with an MEK protein kinase inhibitor, is known in the art and is described in Keipu, L.M. et al., Roigop Etbbot Totodtaru (PET) 1tad §§ Ce11 Progo11Gega11op ίη Oiso1odu, CHtsa1 Oiso1odu, 16: 176-185 (2004), which is hereby incorporated by reference in its entirety. The MEK protein kinase inhibitor (for example, compound A) was studied under ίη νί \ Ό conditions to determine the effect on cancer cell proliferation. This study included 50 volunteers who were diagnosed with pancreatic cancer at a similar stage of development of the malignant neoplasm. A combination of compound A was administered to 25 patients. A placebo was administered to the other 25 patients. Each patient was given a daily daily dose for 14 days with a radioactive indicator, such as labeled fluoro-2-deoxy-ER-glucose (HES).

After treatment for 14 days, a trained specialist using a non-invasive positron emission tomography (PET) device determined the proliferation of tumor cells. In addition, a trained specialist will calculate cell proliferation in both the tumor and normal tissue of patients treated with compound A and placebo. The results indicate a decrease in cell proliferation when treated with a MEK inhibitor of protein kinase (for example, compound A) compared with placebo. This study to determine cell proliferation using radioactively labeled indicators and PET imaging is hereinafter referred to as a method for cell proliferation ίη \ уо. Other methods for proliferating ίη νί \ клеток cells are known in the art.

Similar assays can be used to determine the reduction in tumor size.

Example 125

Study of apoptosis ίη uguo.

The MEK inhibitor, for example compound A, was studied under conditions of ίη V1yo to determine its effect on apoptosis of cancer cells. 40 patients were included in this study voluntarily, all of them were diagnosed with pancreatic cancer at a similar stage of development of malignant neoplasms. Compound A was administered to 20 patients and placebo was administered to 20 patients. Each patient was given a daily dose for 14 days.

After 14 days, each patient used a detectable reagent lipopolysaccharide-binding protein (LBP) associated with the label. In accordance with VΟ 2006/054068, which is thus fully incorporated into this invention by reference, each patient was examined using a scanning device to determine the consumed reagent associated with dead cells. The number of dead cells may correlate with the level of apoptosis in each patient. The levels of apoptosis in patients who were given combinations and those who were given a single drug can be compared with each other, as well as with the control group who were given a placebo. This study to determine apoptosis levels using a lipopolysaccharide-binding protein and a scanning device is hereinafter referred to as apoptosis method способη U1Uo.

Example 126

Dissolution studies.

Capsules containing compound A were prepared as described in the previous examples. Received the following dissolution data using the method of dissolution IP8 <711>.

form I mg form 10 mg Time (min) % release (% Κ5ϋ) % release (% Κ8ϋ) fifteen 78 (8.3) 80 (7.3) thirty 82 (7.1) 87 (9.2) 45 82 (6.7) 92 (9.6) 60 88 (6.3) 92 (7.2) 70 86 (5.7) 95 (5.4)

Claims (16)

CLAIM 1. The crystalline polymorphic form A ^ (-) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide, which has X-ray powder diffraction pattern containing at least 50% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. 2. The crystalline polymorphic form A according to claim 1, wherein the X-ray powder diffraction pattern contains at least 70%, in particular at least 90% of the peaks identified in the X-ray powder diffraction pattern shown in FIG. 5. 3. The crystalline polymorphic form A of claim 1, wherein the X-ray powder diffraction pattern is substantially identical to the X-ray powder diffraction pattern shown in FIG. 5. 4. The crystalline polymorphic form A ^ (-) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide according to claims 1-3, which exhibits a differential scanning calorimetry pattern substantially identical to the differential scanning calorimetry pattern shown in FIG. 6. 5. The crystalline polymorphic form A according to any one of claims 1 to 3 or 4, which has a melting point, which is manifested, as determined using differential scanning calorimetry, at approximately 143 ° C. 6. The crystalline polymorphic form A according to any one of claims 4 or 5, which essentially does not contain water. 7. The crystalline polymorphic form A according to any one of claims 4, 5 or 6, which essentially does not contain a solvent. 8. A pharmaceutical composition for treating a disorder mediated by MEK, which contains the crystalline polymorphic form A according to any one of claims 1 to 7 in an effective amount and at least one excipient or carrier. 9. The crystalline polymorphic form A ^ (-) - (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) -6methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide according to claim 1 obtained by a method comprising the crystallization step of amorphous ^ (3,4-difluoro-2- (2-fluoro-4-iodophenylamino) 6-methoxyphenyl) -1- (2,3-dihydroxypropyl) cyclopropane-1-sulfonamide from mixtures of ethyl acetate and heptane. 10. The use of crystalline polymorphic form A according to any one of claims 1 to 7 and 9 for the preparation of a medicament intended for the treatment of a disorder mediated by MEK in an individual suffering from said disorder. 11. The use of the composition of claim 8 for the preparation of a medicament for the treatment of a disorder mediated by MEK in an individual suffering from the disorder. 12. The use according to claim 10 or 11, where the specified violation mediated by MEK is selected from the group including inflammatory diseases, infections, autoimmune disorders, stroke, ischemia, impaired cardiac activity, neurological disorders, fibrogenetic disorders, proliferative disorders, hyperproliferative disorders, tumors, leukemia, neoplasms, malignant neoplasms, carcinomas, metabolic diseases and malignant diseases. 13. The use of crystalline polymorphic form A according to any one of claims 1 to 7 and 9 for the preparation of a medicinal product intended for the treatment or prevention of proliferative disease in an individual. 14. The use of the composition of claim 8 for the preparation of a medicinal product intended for the treatment or prevention of proliferative disease in an individual. 15. The application of item 13 or 14, where the specified proliferative disease is a malignant neoplasm, psoriasis, restenosis, disease or atherosclerosis. 16. The application of clause 15, where the specified malignant neoplasm is brain cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, colorectal cancer, leukemia, myeloid leukemia, glioblastoma , follicular lymphoma, acute leukemia of B cell precursors, B-cell chronic lymphocytic leukemia, gastric cancer, mesothelioma or small cell lung cancer.