WO2006020564A1

WO2006020564A1 - Pyrimidin derivatives for the treatment of multiple myeloma

Info

Publication number: WO2006020564A1
Application number: PCT/US2005/028118
Authority: WO
Inventors: Rakesh Kumar; Arundathy Nirmalini Pandite
Original assignee: Smithkline Beecham Corporation
Priority date: 2004-08-09
Filing date: 2005-08-09
Publication date: 2006-02-23

Abstract

The present invention relates to a method of treating cancer in a mammal by administration of pyrimidine derivatives and pharmaceutical compositions containing the same. In particular, the method relates to a method of treating multiple myeloma by administration of 5-[[4-[(2,3-Dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl]amino]-2-methylbenzenesulfonamide or salts or solvates thereof.

Description

PYRIMIDIN DERIVATIVES FOR THE TREATMENT OF MULTIPLE

MYELOMA

FIELP OF THE INVENTION

The present invention relates to a method of treating cancer in a mammal by administration of pyrimidine derivatives and pharmaceutical compositions containing the same. In particular, the method relates to a method of treating cancer by administration of 5-[[4-[(2,3-Dimethyl-2H-indazol-6-yl)methylamino]-2- pyrimidinyl]amino]-2-methylbenzenesulfonamide or salts and/or solvates thereof. BACKGROUND OF THE INVENTION

Effective chemotherapy for cancer treatment is a continuing goal in the oncology field. Generally, cancer results from the deregulation of the normal processes that control cell division, differentiation and apoptotic cell death. Apoptosis (programmed cell death) plays essential roles in embryonic development and pathogenesis of various diseases, such as degenerative neuronal diseases, cardiovascular diseases and cancer.

In cancer, the growth of tumors has been shown to be dependent on angiogenesis. The progression of leukemias as well as the accumulation of fluid associated with malignant ascites and pleural effusions also involve pro-angiogenic factors. (See Folkmann, J., J. Nat'l. Cancer Inst., 1990, 82, 4-6.) Consequently, the targeting of pro-angiogenic pathways is a strategy being widely pursued in order to provide new therapeutics in these areas of great, unmet medical need.

Central to the process of angiogenesis are vascular endothelial growth factor (VEGF) and its receptors, termed vascular endothelial growth factor receptor(s)

(VEGFRs). Three PTK receptors for VEGF have been identified: VEGFR1 (FIM )₁

VEGFR2 (FIk--) and KDR), and VEGFR3 (Flt-4). These receptors are involved in angiogenesis and participate in signal transduction. (Mustonen, T. et al J. Cell Biol.

1995:129:895-898; Ferrara and Davis-Smyth, Endocrine Reviews, 18(1 ):4-25, 1997; McMahon, G., The Oncologist, Vol. 5, No 90001 , 3-10, April 2000).

Multiple myeloma (MM) is a malignant plasma cell disorder accounting for approximately 10% of hematologic malignancies (Blade, J. et al. British Journal of Haematology, 1998: 102: 11 15-23). Emerging evidence suggests that VEGF may play an important role in the pathogenesis of MM, Myeloma cells express VEGFR-1 , and VEGF has been shown to induce proliferation and migration of the malignant cell (Podar, K. et al. Journal Bioligical Chemistry, 2002: 277: 7875-7881). Elevated serum levels of VEGF and basic hepatocyte growth factor (HGF) have been reported in patients with MM (Iwasaki, T et al. British Journal of Haematology, 2002: 116: 796- 802). Furthermore, VEGF is expressed and secreted by myeloma cell lines and patient cells, as well as by bone marrow stromal cells (BMSCs).

Consequently, antagonism of the VEGFR kinase domain is believed to block phosphorylation of tyrosine residues and serve to disrupt initiation of angiogenesis, and other signaling pathways mediated by VEGF, thereby providing a potent treatment for cancers such as multiple myeloma. SUMMARY OF THE INVENTION

The present inventors have now identified novel cancer treatment methods which include administration of 5-[[4-[(2,3-Dimethyl-2H-indazol-6-yl)methylamino]-2- pyrimidinyl]amino]-2-methylbenzenesulfonamide as well as salts and/or solvates thereof.

In one aspect of the present invention, there is provided a method of treating multiple myeloma in a mammal, comprising: administering to the mammal a compound of formula (I)

or a salt or solvate thereof. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is a representative experiment demonstrating the effect of the compound of Example 1 on Flt-1 phosphorylation and VEGF- triggered activation of downstream signaling molecules in MM.1 S cells.

Figure 2 is a representative experiment demonstrating the effects of the compound of Example 1 on multiple myeloma cell survival, and expression of caspases and apoptosis- associated proteins. Figure 3 is a representative experiment demonstrating the effect of the compound of Example 1 on CD138+ multiple myeloma patient cells.

Figure 4 is a representative experiment demonstrating the effect of the compound of Example 1 on VEGF-induced signaling pathways in HUVEC cells, and on HUVEC cell survival.

Figure 5 is a representative experiment demonstrating the effect of the compound of Example 1 on VEGF- induced HUVEC-MM cell adhesion and cell proliferation.

Figure 6 is a representative experiment demonstrating the effect of the compound of Example 1 on the sensitivity of MM cells to low-dose DNA-damaging chemotherapeutic agents, alone and when bound to HUVEC and BMSC. DETAILED DESCRIPTION OF THE INVENTION

As used herein the term "neoplasm" refers to an abnormal growth of cells or tissue and is understood to include benign, i.e., non-cancerous growths, and malignant, i.e., cancerous growths. The term "neoplastic" means of or related to a neoplasm.

As used herein, the term "effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for instance, by a researcher or clinician. Furthermore, the term "therapeutically effective amount" means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder. The term also includes within its scope amounts effective to enhance normal physiological function. As used herein, the term "solvate" refers to a complex of variable stoichiometry formed by a solute (in this invention, compounds of formula (I) or a salt thereof) and a solvent. Such solvents for the purpose of the invention may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol and acetic acid. Preferably the solvent used is a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include, without limitation, water, ethanol and acetic acid. Most preferably the solvent used is water. The methods of cancer treatment disclosed herein include administering a compound of formula (I):

or a salt or solvate thereof. In one embodiment, the salt of the compound of formula (I) is a hydrochloride salt. In a preferred embodiment, the salt of the compound of formula (I) is a monohydrochloride salt as illustrated by formula (I¹). The monohydrochloride salt of the compound of formula (I) has the chemical name 5-[[4-[(2,3-Dimethyl-2H-indazol-6- yl)methylamino]-2-pyrimidinyl]amino]-2-methylbenzenesulfonamide monohydrochloride.

(I¹)

In another preferred embodiment, the the salt of the compound of formula (I) is a monohydrochloride monohydrate solvate of the compound of formula (I). The monohydrochloride monohydrate solvate of the compound of formula (I) has the chemical name 5-({4-[(2,3-dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl}amino)- 2-methylbenzenesulfonamide monohydrochloride monohydrate, as illustrated in formula (I").

(I")

The free base, salts and solvates of the compound of formula (I) may be prepared, for example, according to the procedures of International Patent Application No. PCT/US01/49367 filed December 19, 2001 , and published as WO 02/059110 on

August 1 , 2002, and International Patent Application No. PCT/US03/19211 filed June

17, 2003, and published as WO 03/106416 on December 24, 2003.

Typically, the salts of the present invention are pharmaceutically acceptable salts. Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention. Salts of the compounds of the present invention may comprise acid addition salts derived from a nitrogen on a substituent in a compound of the present invention. Representative salts include the following salts: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, monopotassium maleate, mucate, napsylate, nitrate, N-methylglucamine, oxalate, pamoate (embonate), palmitate, pantothenate, phosphate/diphosphate, polygalacturonate, potassium, salicylate, sodium, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, trimethylammonium and valerate. Other salts, which are not pharmaceutically acceptable, may be useful in the preparation of compounds of this invention and these form a further aspect of the invention.

While it is possible that, for use in the cancer treatment methods of the present invention, a compound of formula (I) as well as salts or solvates thereof, may be administered as the raw chemical, it is possible to present the active ingredient as a pharmaceutical composition. Accordingly, the invention further provides pharmaceutical compositions, which may be administered in the cancer treatment methods of the present invention. The pharmaceutical compositions include a compound of formula (I) and salts or solvates thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients. The carrier(s), diluent(s) or excipient(s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

Pharmaceutical formulations may be presented in unit dose forms containing a predetermined amount of active ingredient per unit dose. Such a unit may contain, for example, 0.5mg to 1g, preferably 1 mg to 700mg, more preferably 5mg to 100mg of a compound of formula (I), depending on the condition being treated, the route of administration and the age, weight and condition of the patient, or pharmaceutical formulations may be presented in unit dose forms containing a predetermined amount of active ingredient per unit dose. Preferred unit dosage formulations are those containing a daily dose or sub-dose, as herein above recited, or an appropriate fraction thereof, of an active ingredient. Furthermore, such pharmaceutical formulations may be prepared by any of the methods well known in the pharmacy art.

The compound of formula (I) may be administered by any appropriate route. Suitable routes include oral, rectal, nasal, topical (including buccal and sublingual), vaginal, and parenteral (including subcutaneous, intramuscular, intraveneous, intradermal, intrathecal, and epidural). It will be appreciated that the preferred route may vary with, for example, the condition of the recipient of the combination.

The method of the present invention may also be employed with other therapeutic methods of cancer treatment. In particular, in anti-neoplastic therapy, combination therapy with other chemotherapeutic, hormonal, antibody agents as well as surgical and/or radiation treatments other than those mentioned above are envisaged. Anti-neoplastic therapies are described for instance in International Application No. PCT US 02/01130, filed January 14, 2002, published as WO 02/056912 on July 25, 2002. Combination therapies according to the present invention thus include the administration of at least one compound of formula (I) as well as optional use of other therapeutic agents including other anti-neoplastic agents. Such combination of agents may be administered together or separately and, when administered separately this may occur simultaneously or sequentially in any order, both close and remote in time. The amounts of the compound of formula (I) and the other pharmaceutically active agent(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect.

Pharmaceutical formulations adapted for oral administration may be presented as discrete units such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or oil-in-water liquid emulsions or water-in-oil liquid emulsions.

For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing and coloring agent can also be present. Capsules are made by preparing a powder mixture as described above, and filling formed gelatin sheaths. Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation. A disintegrating or solubilizing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant and pressing into tablets. A powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or an absorption agent such as bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil. The lubricated mixture is then compressed into tablets. The compounds of the present invention can also be combined with free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps. A clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.

Oral fluids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound in a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.

Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The formulation can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material in polymers, wax or the like. The agents for use according to the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

Agents for use according to the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels. Pharmaceutical formulations adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. For example, the active ingredient may be delivered from the patch by iontophoresis as generally described in Pharmaceutical Research, 3(6), 318 (1986).

Pharmaceutical formulations adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.

For treatments of the eye or other external tissues, for example mouth and skin, the formulations are preferably applied as a topical ointment or cream. When formulated in an ointment, the active ingredient may be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredient may be formulated in a cream with an oil-in-water cream base or a water-in-oil base.

Pharmaceutical formulations adapted for topical administrations to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent.

Pharmaceutical formulations adapted for topical administration in the mouth include lozenges, pastilles and mouth washes.

Pharmaceutical formulations adapted for rectal administration may be presented as suppositories or as enemas.

Pharmaceutical formulations adapted for nasal administration wherein the carrier is a solid include a coarse powder having a particle size for example in the range 20 to 500 microns which is administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close up to the nose. Suitable formulations wherein the carrier is a liquid, for administration as a nasal spray or as nasal drops, include aqueous or oil solutions of the active ingredient.

Pharmaceutical formulations adapted for administration by inhalation include fine particle dusts or mists that may be generated by means of various types of metered dose pressurised aerosols, nebulizers or insufflators.

Pharmaceutical formulations adapted for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations.

Pharmaceutical formulations adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets. It should be understood that in addition to the ingredients particularly mentioned above, the formulations may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.

As indicated, a specific compound of formula (I) is administered to a mammal. Typically, the amount of one of the administered agents of the present invention will depend upon a number of factors including, for example, the age and weight of the mammal, the precise condition requiring treatment, the severity of the condition, the nature of the formulation, and the route of administration. Ultimately, the amount will be at the discretion of the attendant physician or veterinarian. Typically, the compound of formula (I) will be given in the range of 0.1 to 100 mg/kg body weight of recipient (mammal) per day and more usually in the range of 1 to 10 mg/kg body weight per day.

As recited above the present invention includes cancer treatment methods through administration of 5-[[4-[(2,3-Dimethyl-2H-indazol-6-yl)methylamino]-2- pyrimidinyl]amino]-2-methylbenzenesulfonamide or salts or solvates thereof.

In a preferred embodiment of the present invention, there is provided a method of treating multiple myeloma in a mammal, comprising: administering to the mammal a compound of formula (I) or salts or solvates thereof. In another preferred embodiment, the compound is a compound of formula (I¹). In another preferred embodiment, the compound is a compound of formula (I").

In one embodiment of the present invention, there is provided a method of treating multiple myeloma in a mammal, comprising: administering to the mammal a compound of formula (I) or salts or solvates thereof and at least one anti-neoplastic therapy. In a preferred embodiment, the compound is a compound of formula (I'). In another preferred embodiment, the compound is a compound of formula (I").

In the foregoing cancer treatment methods of the present invention the compounds of formulae (I), (I') and (I") are as described above.

The following examples are intended for illustration only and are not intended to limit the scope of the invention in any way. EXAMPLES

As used herein the symbols and conventions used in these processes, schemes and examples are consistent with those used in the contemporary scientific literature, for example, the Journal of the American Chemical Society or the Journal of Biological Chemistry. Standard single-letter or three-letter abbreviations are generally used to designate amino acid residues, which are assumed to be in the L-configu ration unless otherwise noted. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification. Specifically, the following abbreviations may be used in the examples and throughout the specification: g (grams); mg (milligrams);

L (liters); ml_ (milliliters); μL (microliters); psi (pounds per square inch);

M (molar); mM (millimolar);

N (Normal) Kg (kilogram) i. v. (intravenous); Hz (Hertz);

MHz (megahertz); mol (moles); mmol (millimoles); RT (room temperature); min (minutes); h (hours); mp (melting point); TLC (thin layer chromatography);

Tr (retention time); RP (reverse phase);

DCM (dichloromethane); DCE (dichloroethane); DMF

(Λ/,Λ/-dimethylformamide); HOAc (acetic acid);

TMSE (2-(trimethylsilyl)ethyl); TMS (trimethylsilyl);

TIPS (triisopropylsilyl); TBS (f-butyldimethylsilyl);

HPLC (high pressure liquic I chromatography);

THF (tetrahydrofuran); DMSO (dimethyisulfoxide);

EtOAc (ethyl acetate); DME (1 ,2-dimethoxyethane);

EDTA ethylenediaminetetraacetic acid

FBS fetal bovine serum

IMDM Iscove's Modified Dulbecco's medium

PBS phosphate buffered saline

RPMI Roswell Park Memorial Institute

RIPA buffer _*

RT room temperature ^*150 mM NaCI, 50 mM Tris-HCI, pH 7.5, 0.25% (w/v) -deoxycholate, 1 % NP-40, 5 mM sodium orthovanadate, 2 mM sodium fluoride, and a protease inhibitor cocktail.

Unless otherwise indicated, all temperatures are expressed in ⁰C (degrees Centigrade). All reactions conducted under an inert atmosphere at room temperature unless otherwise noted.

In the following examples and figures, recitation of "GW786034" or "GW78" denotes a compound of formula (I) or a salt or solvate thereof.

The following examples describe the syntheses of intermediates particularly useful in the synthesis of compounds of Formula (I): Intermediate Example 1

Preparation of 2,3-dimethyl-6-nitro-2H-indazole

Procedure 1 :

To a stirred solution of 18.5 g (0.11 mol) of 3-methyl-6-nitro- 7H-indazole in 350 ml acetone, at room temperature, was added 20 g (0.14 mol) of trimethyloxonium tetraflouroborate. After the solution was allowed to stir under argon for 3 hours, the solvent was removed under reduced pressure. To the resulting solid was added saturated aqueous NaHCO₃ (600 ml_) and a 4:1 mixture of chloroform-isopropanol

(200 ml), the mixture was agitated and the layers were separated. The aqueous phase was washed with additional chloroform: isopropanol (4 x 200 mL) and the combined organic phase was dried (Na₂SO₄). Filtration and removal of solvent gave a tan solid.

The solid was washed with ether (200 mL) to afford 2,3-dimethyl-6-nitro-2H-indazole as a yellow solid (15.85 g, 73 %). ¹H NMR (300 MHz, DMSO-d₆) δ 8.51 (s, 1 H), 7.94 (d, J

= 9.1 Hz, 1 H), 7.73 (d, J = 8.9 Hz, 1 H), 4.14 (s, 3H)₁ 2.67 (s, 3H). MS (ES+, m/z) 192 (M+H).

Procedure 2:

Trimethyl orthoformate (11 mmol, 1.17 g) was added over a 2 min period to a solution of boron trifluoride etherate (12.5 mmol, 1.77 g in methylene chloride (2.0 mL) which had been cooled to -30 ⁰C. The mixture was warmed to 0 ⁰C for 15 min and was then cooled to -70 ⁰C. The nitro indazole (10 mmol, 1.77 g) was slurried in methylene chloride (30 ml_) and was added all at once to the cooled mixture. The mixture was stirred at -70 ⁰C for 15 min and at ambient temperature for 17 h. After 17 h the mixture was red and heterogeneous. The reaction mixture was quenched with saturated sodium bicarbonate solution (20 ml_) and the organic layer separated. The aqueous layer was extracted with methylene chloride (30 ml_). The methylene chloride layers were combined and extracted with water (30 ml_). The methylene chloride layer was distilled under reduced pressure until ~ 10 ml_ remained. Propanol (10 ml_) was added and the remainder of the methylene chloride removed under reduced pressure, resulting in a yellow slurry. The product was isolated by filtration to give 2,3-dimethyl-6- nitro-2H-indazole (65 %, 7mmol, 1.25 g) as a light yellow powder. ¹H NMR (300 MHz, DMSO-d₆) δ 8.51 (s, 1 H), 7.94 (d, J = 9.1 Hz, 1 H), 7.73 (d, J = 8.9 Hz₁ 1 H), 4.14 (s, 3H), 2.67 (s, 3H). MS (ES+, m/z) 192 (M+H). Procedure 3: In a 25 ml round bottom flask 3-methyl-6-nitroindazole (7.27 mmol, 1.28 g) was dissolved with stirring in DMSO (4.0 mL) and was treated with concentrated sulfuric acid (7.27 mmol, 0.73 g) to yield a thick slurry. The slurry was treated with dimethyl sulfate (21.1 mmol, 2.66 g). The mixture was heated under nitrogen at 50 ⁰C for 72 h. After 72 h a thick yellow slurry was obtained. The slurry was cooled and was slowly treated with saturated sodium bicarbonate solution (10 mL). The mixture was extracted with methylene chloride (2 x 20 mL). The methylene chloride layers were combined and back extracted with water (20 mL). The methylene chloride layer was treated with propanol (10 mL) and the methylene chloride was removed by distillation under reduced pressure. The solid was isolated by filtration and the yellow solid washed with heptane (5 mL) and air-dried. The 2,3-dimethyl-6-nitro-2H-indazole product (70%, 0.97 g) was obtained as a light yellow solid. ¹H NMR (300 MHz, DMSO-d₆) δ 8.51 (s, 1 H), 7.94 (d, J = 9.1 Hz, 1 H), 7.73 (d, J= 8.9 Hz, 1 H), 4.14 (s, 3H), 2.67 (s, 3H). MS (ES+, m/z) 192 (M+H).

Procedure 4: Into a 250 mL 3-necked round bottom flask was placed 3-methyl-6-nitro-1 H- indazole sulfuric acid salt (5.0 g, 18.2 mmol) and methylene chloride (25 mL). The mixture was stirred at 25 ⁰C and was treated with DMSO (5 mL). Dimethyl sulfate (6.7 g, 5.0 mL, 53.0 mmol) was added via syringe and the reaction was heated at reflux in a 70 ⁰C bath. After 7 h HPLC analysis showed 9% starting material. At this point heating was stopped and the workup begun. Saturated sodium bicarbonate solution (35 ml_) was added to the reaction mixture at RT. The layers were allowed to separate and the aqueous layer was extracted with methylene chloride (25 ml_). The methylene chloride layers were combined and washed with water (2 x 25 ml_). The methylene chloride layer was distilled under reduced pressure until half the volume was removed. Propanol (25 ml_) was added and distillation under reduced pressure was continued until all the methylene chloride had been removed. This yielded a yellow slurry, which was allowed to stir at 25 ⁰C for 1 h. The product was isolated via filtration and the resulting yellow solid was washed with heptane (10 ml_). This yielded 2,3-dimethyl-6- nitro-2H-indazole (70%, 2.43 g) as a yellow solid. ¹H NMR (300 MHz, DMSO-d₆) δ 8.51 (s, 1 H), 7.94 (d, J = 9.1 Hz, 1 H)₁ 7.73 (d, J = 8.9 Hz, 1 H), 4.14 (s, 3H), 2.67 (s, 3H). MS (ES+, m/z) 192 (M+H). Intermediate Example 2 Preparation of2,3-dimethyl-6-amino-2H-indazole

Procedure 1 :

To a stirred solution of 2,3-dimethyl-6-nitro-2H-indazole (1.13 g) in 2- methoxyethyl ether (12 ml), at 0 ⁹C, was added a solution of 4.48 g of tin(ll) chloride in

8.9 ml of concentrated HCI dropwise over 5 min. After the addition was complete, the ice bath was removed and the solution was allowed to stir for an additional 30 min.

Approximately 40 ml of diethyl ether was added to reaction, resulting in precipitate formation. The resulting precipitate was isolated by filtration and washed with diethyl ether, and afforded a yellow solid (1.1 g, 95 %), the HCI salt 2,3-dimethyl-2H-indazol-

6-amine. ¹H NMR (300 MHz, DMSO-d₆) δ 7.77 (d, J = 8.9 Hz, 1 H), 7.18 (s, 1 H), 7.88

(m, 1 H), 4.04 (s, 3H), 2.61 (s, 3H). MS (ES+, m/z) 162 (M+H).

Procedure 2: A 2-L 3-necked round bottom flask was fitted with nitrogen inlet and outlet and with mechanical stirring. A moderate nitrogen flow was initiated and the reactor was charged with 10 % Pd/C (50% water wet, 6.0 g). Stirring was initiated and the reactor was charged with methanol (750 mL) and the product of Intermediate Example 1 (50 g). Ammonium formate (82.54 g) was dissolved in water (120 mL). The water solution of ammonium formate was added to the reaction solution at an addition rate, which kept the reaction temperature at or between 25 and 30 ⁰C. The reaction was allowed to proceed at 25 ⁰C. After 6 h the reaction was judged to be finished based on HPLC analysis. The mixture was filtered and the catalyst washed with methanol (50 mL). The methanol layers were combined and the solvent removed under reduced pressure. The residue was dissolved in water (200 mL) and was extracted with methylene chloride (3 x 250 mL). The methylene chloride layers were combined and solvent removed under vacuum to remove approximately half the solvent. Heptane (400 mL) was added and the vacuum distillation continued until approximately 300 mL reaction product slurry remained. The product was isolated by filtration and dried under vacuum at 50 ⁰C for 4 h. to yield 2,3-dimethyl-6-amino-2H-indazole as the free base. (40.76 g, 96.7 %). ¹H NMR (300 MHz, DMSO-d₆) δ 7.31 (d, J = 8.9 Hz, 1 H), 6.45 (d, J = 8.9 Hz, 1 H), 6.38 (s, 1 H), 4.95 (s, br, 2H), 3.85 (s, 3H), 2.44 (s, 3H) MS (ES+, m/z) 162 (M+H). Intermediate Example 3 Preparation of N-(2-chloropyrimidin-4-yl)-2,3-dimethyl-2H-indazol-6-amine

Procedure 1

To a stirred solution of the product of Intermediate Example 2 (2.97 g, .015 mol) and NaHCO₃ (5.05 g, .06 mol) in THF (15 mL) and ethanol (60 mL) was added 2,4- dichloropyrimidine (6.70 g, .045 mol) at rt. After the reaction was stirred for four hours at 85 ⁰C, the suspension was cooled to rt., filtered and washed thoroughly with ethyl acetate. The filtrate was concentrated under reduced pressure, and the resulting solid was triturated with ethyl acetate to yield Λ/-(2-chloropyrimidin-4-yl)-2,3-dimethyl-2W- indazol-6-amine (89 %, 3.84 g). ¹H NMR (400 MHz, DMSO-d₆) δ 7.28 (d, J = 9.0 Hz, 1 H), 6.42 (d, J = 8.8 Hz, 1 H), 6.37 (s, 1 H), 5.18 (br s, 1 H), 3.84 (s, 3H), 2.43 (s, 3H). MS (ES+, m/z) 274 (M+H). Procedure 2

To a 1-L 3-necked flask equipped with air-driven mechanical stirrer, thermometer, and nitrogen inlet/outlet was charged a solution of the product of Intermediate Example 2 (32.89 g, 0.204 mol, 1.0 equiv) in 425 ml_ (13 volumes) of EtOH/THF (4/1 ), sodium bicarbonate (51.42 g, 0.612 mol, 3.0 equiv) and then 2,4- dichloropyrimidine (45.59 g, 0.306 mol, 1.5 equiv). The flask contents were heated to

75 ⁰C and held at 74 - 76 ⁰C for 6 - 7 hrs. The progress of the reaction was checked by HPLC (the product of Intermediate Example 2 < 2%). The reaction contents were cooled to 20 - 25 ⁰C over 30 min, and kept at 20 - 25 ⁰C for 30 min. Then the reaction contents were further cooled to 10 - 12 ⁰C over 30 min, and kept at that temperature for an additional 10 min. The contents were filtered and filter cake washed with EtOAc

(2 x 100 ml_, 3.0 volumes), and deionized water (514 ml_, 15.6 volumes). The filter cake was then dried in a vacuum oven at 35 ⁰C overnight to afford the desired product 44.75 g as a white solid (80.1 %). ¹H NMR (400 MHz, DMSO-d₆) δ 7.28 (d, J = 9.0 Hz, 1 H), 6.42 (d, J = 8.8 Hz, 1 H), 6.37 (s, 1 H), 5.18 (br s, 1 H), 3.84 (s, 3H), 2.43 (s, 3H). MS (ES+, m/z) 274 (M+H). Intermediate Example 4

Preparation of N-(2-chloropyrimidin-4-yl)-N,2, 3-trimethyl-2H-inda∑ol-6-amine

Procedure 1 To a stirred solution of the product of Intermediate Example 3 (7.37 g) in DMF

(50 ml) was added Cs₂CO₃ (7.44 g, 2 eqv.) and iodomethane (1.84 ml, 1.1 eqv.) at room temperature. The mixture was stirred at rt overnight. The reaction mixture was then poured into an ice-water bath, and the precipitate was collected via filtration and washed with water. The precipitate was air-dried to afford Λ/-(2-chloropyrimidin-4-yl)- /V,2,3-trimethyl-2H-indazol-6-amine as an off-white solid (6.43 g, 83%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.94 (d, J = 6.0 Hz, 1 H), 7.80 (d, J = 7.0 Hz, 1 H), 7.50 (d, J = 1.0 Hz, 1 H), 6.88 (m, 1 H), 6.24 (d, J = 6.2 Hz, 1 H), 4.06 (s, 3H), 3.42 (s, 3H), 2.62 (s, 3H). MS (ES+, m/z) 288 (M+H). Procedure 2

A 3L 3-necked flask equipped with air-driven mechanical stirrer, thermometer, addition funnel and nitrogen inlet/outlet was charged with DMF (272 ml_, 5 volumes) and the product of Intermediate Example 3 (54.4 g, 0.20 mol, 1.0 equiv) with stirring. The reaction mixture was further charged with cesium carbonate (194.5 g, 0.60 mol, 3.0 equiv) while maintaining the reaction temperature between 20 ~ 25 ⁰C. The reaction mixture was stirred at 20 ~ 25 ⁰C for 10 minutes, lodomethane (45.1 g, 0.32 mol, 1.6 equiv) was charged over ~ 10 minutes while maintaining the temperature 20 ~ 3O⁰C. The reaction mixture was stirred at 20 ~ 30 ⁰C (Typically, the reaction is complete in 1 ~ 2 hours). Deionized H₂O (925 ml_, 17 volumes) was added over ~ 30 minutes while maintaining the temperature at 25 ~ 40 ⁰C. The reaction mixture was stirred at 20 ~ 25 ⁰C for 40 minutes. The product was isolated by filtration and then the filter cake washed with H₂O / DMF (6 : 1 , 252 ml_, 4.6 volumes). The wet cake was dried under vacuum at 40 ~ 45 ⁰C and Λ/-(2-chloropyrimidin-4-yl)-Λ/,2,3-trimethyl-2H- indazol-6-amine (51.7 g, 90.4%) was isolated as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 7.94 (d, J = 6.0 Hz, 1 H), 7.80 (d, J = 7.0 Hz, 1 H), 7.50 (d, J = 1.0 Hz, 1 H), 6.88 (m, 1 H), 6.24 (d, J = 6.2 Hz, 1 H), 4.06 (s, 3H), 3.42 (s, 3H), 2.62 (s, 3H). MS (ES+, m/z) 288 (M+H). Example 1

5-({4i(2,3-dimethyl-2H-indazol-6-yl)(methyl)amino]pyrimidin-2-yl}am methylbenzenesulfonamide

Procedure 1

To a solution of Intermediate Example 4 (200 mg, 0.695 mmol) and 5-amino-2- methylbenzenesulfonamide (129.4 mg, 0.695 mmol) in isopropanol (6 ml) was added 4 drops of cone. HCI. The mixture was heated to reflux overnight. The mixture was cooled to rt and diluted with ether (6 ml). Precipitate was collected via filtration and washed with ether. The hydrochloride salt of 5-({4-[(2,3-dimethyl-2H-indazol-6- yl)(methyl)amino]-pyrimidin-2-yl}amino)-2-methylbenzenesulfonamide was isolated as an off-white solid. ¹H NMR (400 MHz, d₆DMSO+NaHCO₃) δ 9.50 (br s, 1 H), 8.55 (br s, 1 H), 7.81 (d, J = 6.2 Hz, 1 H), 7.75 (d, J = 8.7 Hz, 1 H), 7.69 (m, 1 H), 7.43 (s, 1 H), 7.23 (s, 2H), 7.15 (d, J = 8.4 Hz, 1 H), 6.86 (m, 1 H), 5.74 (d, J = 6.1 Hz, 1 H), 4.04 (s, 3H), 3.48 (s, 3H), 2.61 (s, 3H), 2.48 (s, 3H). MS (ES+, m/z) 438 (M+H). Procedure 2

A 250-mL 3-necked flask equipped with a magnetic stir bar, thermometer, reflux condenser, and nitrogen inlet/outlet was charged with ethanol (60 mL, 10 volumes), the product of Intermediate Example 4 (6.00 g, 20.85 mmol, 1.0 equiv) and 5-amino-2- methylbenzenesulfonamide (4.00 g, 21.48 mmol, 1.03 equiv) with stirring. The reaction mixture was heated to 70 ⁰C. After stirring the reaction mixture at 68 - 72 ⁰C for 3 hrs, 4M HCI in dioxane (0.11 mL, 0.44 mmol, 0.02 equiv) was charged over ca. 2 min. The reaction mixture was stirred at 68 - 72 ⁰C until < 1.5% by area of the starting product of Intermediate Example 4 was remaining by HPLC analysis (Typically, this reaction is complete in > 8 hrs). The reaction mixture was cooled to 20 ⁰C over ca. 30 min and stirred at 20 - 22 ⁰C for 40 min. The product was then isolated by filtration and the filter cake washed with ethanol (20 mL, 3.3 volumes). The wet cake was dried under vacuum at 45 - 50 ⁰C. The monohydrochloride salt of 5-({4-[(2,3-dimethyl-2/-/-indazol- 6-yl)(methyl)amino]-pyrimidin-2-yl}amino)-2-methylbenzenesulfonamide (9.52 g, 96.4%) was isolated as a white solid. ¹H NMR (400 MHz, d₆DMSO+NaHCO₃) δ 9.50 (br s, 1 H), 8.55 (br s, 1 H), 7.81 (d, J = 6.2 Hz, 1 H), 7.75 (d, J = 8.7 Hz, 1 H), 7.69 (m, 1 H), 7.43 (s, 1 H), 7.23 (s, 2H), 7.15 (d, J = 8.4 Hz₁ 1 H), 6.86 (m, 1 H)₁ 5.74 (d, J = 6.1 Hz, 1 H), 4.04 (s, 3H), 3.48 (s, 3H), 2.61 (s, 3H)₁ 2.48 (s, 3H). MS (ES+, m/z) 438 (M+H). Procedure 3:

To a stirred suspension of the product of Intermediate Example 4 (1 .1 g, 3.8 mmol) in 14 mL of MeOH, was added 5-amino-2-methylbenzenesulfonamide (0.78 g,

4.2 mmol, 1.1 equiv) at room temperature. The reaction mixture was heated at reflux for 3 h, then 4 M HCI in 1 ,4-dioxane (19 μl_, 0.076 mmol) was added in one portion.

After 4 h, the suspension was cooled to room temperature, and filtered. The resulting solid was washed with 10 mL of MeOH and dried in vacuo to yield 1 .3 g (72%) of 5-({4- [(2,3-dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl}amino)-2-methyl benzenesulfonamide monohydrochloride as a white solid. ¹ H NMR (DMSO-d6, 400 MHz) δ 10.95 (s, 1 H), 8.36 (s, 1 H), 7.86 (d, J = 8.8 Hz, 2H), 7.64-7.59 (m, 2H), 7.40 (m, 3H), 6.93 (dd, J = 8.8, 2.0 Hz, 1 H), 5.92 (s, 1 H), 4.08 (s, 3H), 3.57 (s, 3H), 2.65 (s, 3H), 2.56 (s, 3H). Procedure 4

To a stirred suspension of the product of Intermediate Example 4 (1 .1 g, 3.7 mmol) in 10 mL of THF, was added 5-amino-2-methylbenzenesulfonamide (0.70 g, 3.8 mmol, 1 .0 equiv) at room temperature. The reaction mixture was heated at reflux for 3 h, then 4 M HCI in 1 ,4-dioxane (18 μL, 0.072 mmol) was added in one portion. After 5 h, the suspension was cooled to room temperature, and filtered. The resulting solid was washed with 16 mL of THF and dried in the air to yield 1 .6 g (92%) of 5-({4-[(2,3- dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl}amino)-2-methylbenzene sulfonamide monohydrochloride as a light yellow solid. Procedure 5 To a stirred suspension of the product of Intermediate Example 4 (1 .0 g, 3.6 mmol) in 10 mL of CH3CN, was added 5-amino-2-methylbenzenesulfonamide (0.70 g,

3.8 mmol, 1 .Oequiv) at room temperature. The reaction mixture was heated at reflux for

3 h, then 4 M HCI in 1 ,4-dioxane (18 μL, 0.076 mmol) was added in one portion. After

20 h, the suspension was cooled to room temperature, and filtered. The resulting solid was washed with 10 mL of CH3CINI and dried in the air to yield 1.3 g (73%) of 5-({4-

[(2,3-dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl}amino)-2-methyl benzenesulfonamide monohydrochloride as an off-white solid. Procedure 6:

Preparation of 5-({4-[(2,3-dimethyl-2H~indazol-6-yl)methylamino]-2- pyrimidinyl}amino)-2-methylbenzenesulfonamide monohydrochloride monohydrate.

To a round bottom flask, was added 2.6 g of the monohydrochloride salt of Example 1 , procedure 1 , any form. Then added was 39 mL of isopropanol (15 volumes). The mixture was heated to 75 deg C in an oil bath, then 14 mL of 0.05N aqueous HCI (5.4 volumes) was added. The clear solution was cooled to 65 deg C, then seeded with the monohydrate of the monohydrochloride salt of Example 1 , procedure 1 (0.05-0.1 wt %). The cloudy solution was stirred at 65 deg C for 60 minutes, then cooled to 0 deg C at -0.25-0.5 deg C/min. The resulting white solid was filtered and dried to constant weight under vacuum at RT to give 88% yield of 5 -({4- [(2,3-dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl}amino)-2-methylbenzene sulfonamide monohydrochloride monohydrate. Biological Data Figure 1.

Effect of the compound of Example 1 on FIt-I phosphorylation and VEGF- triggered activation of downstream signaling molecules, (a) MM.1 S cells were starved overnight in RPMM 640 with 1 % FBS and for 3h in RPMI 1640 with no FBS. After pretreatment with the compound of Example 1 (1 h; 10μg/mL) or DMSO, MM.1 S cells were stimulated with 100ng/ml_ VEGF for indicated intervals. FIM immunoprecipitates from whole cell lysates were analyzed by western blotting using antisera against phosphotyrosine residues. Equal loading was confirmed by immunoblotting with antisera directed against FIM . (b) MM.1 S cells pretreated as in panel a were stimulated with 10Ong/mL VEGF in the presence or absence of the compound of Example 1 for 1 , 5, 30 min. Whole cell lysates were analyzed by westen blotting using antisera against phospho-Akt-1 (pAkti). Immunoblotting for Akt1 confirmed equal protein loading. Results are as shown in Figure 1. Figure 2. Effect of the compound of Example 1 on MM cell survival, caspases and apoptosis- associated proteins, (a) Dose-related effect of the compound of Example 1 on survival. MM cell lines were cultured with control media or with the compound of Example 1 for 48h. MTT cleavage was measured during the last 8h of 48h cultures. (b) Protein profiling of MM cells exposed to the compound of Example 1. MM.1 S cells were exposed to the compound of Example 1 for indicated intervals, followed by immunoblot analysis of the lysates with indicated antibodies. Results are as shown in Figure 2. Figure 3.

Effect of the compound of Example 1 on CD138+ MM patient cells. Unpurified bone marrow (BM) mononuclear cells from BM aspirate of MM patient (which contain CD138+ MM cells and CD138- normal cells of the bone microenvironment, such as stromal cells) were cultured in the presence of 10% FBS for 5 days. Results are as shown in Figure 3. Figure 4.

Effect of the compound of Example 1 on VEGF-induced signaling pathways in HUVEC cells, and on cell survival. Prior to treatment, cells were starved in EGM-2 with 2% FBS overnight, followed by 3h in EGM-2 without FBS. Cells were then treated for 1 h with 10 μg/ml of the compound of Example 1 , or DMSO as a control. Following treatment, the cells were stimulated with 100ng/mL of VEGF, as shown in Figure 4. Figure 5. Effect of the compound of Example 1 on VEGF- induced HUVEC-MM cell adhesion.

(a) HUVECs were starved overnight in EGM-2 with 2% FBS. HUVECs were then stimulated with 100ng/mL VEGF for indicated intervals. Whole cell lysates were analyzed by westen blotting using antisera against ICAM-1 and VCAM-1. lmmunoblotting for actin confirmed equal protein loading.

(b) Dose-dependent inhibition of VEGF- induced MM cell adhesion on HUVECs. (c) The compound of Example 1 inhibits proliferation of MM cells adherent to HUVECs. MM cells were cultured with or without HUVECs. The compound of Example 1 was added in the indicated concentrations, and proliferation was measured using [³H] thymidine-uptake. Data shown are mean +/- SD of experiments performed in triplicate. Results are as shown in Figure 5. Figure 6. Effects of the compound of Example 1 on sensitivity of MM cells alone and on tumor cells bound to HUVEC and BMSC, to low-dose DNA-damaging chemotherapeutic agents, (a) MM.1 S cells were left untreated or treated with the compound of Example 1 (1 and 5μg/mL). IMiDI (1μM), melphalan (10μM), and bortezomib (PS341) (5nM), were added where indicated. MTT cleavage was measured during the last 8h of 24h and 48h cultures, (b-c) MM-HUVEC co-cultures were left untreated or treated with the indicated concentrations of the compound of Example 1. IMiDI or melphalan were added where indicated, at the concentrations as in (a). Proliferation was measured using [³H] thymidine-uptake. Results are as shown in Figure 6.

Although specific embodiments of the present invention are herein illustrated and described in detail, the invention is not limited thereto. The above detailed descriptions are provided as exemplary of the present invention and should not be construed as constituting any limitation of the invention. Modifications will be obvious to those skilled in the art, and all modifications that do not depart from the spirit of the invention are intended to be included with the scope of the appended claims.