Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
  • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/08—Antiallergic agents

Definitions

• the present invention relates to a new use of the N-phenyl-2-pyrimidine-amine derivatives of formula I in which the symbols and substituents have the meaning as given hereinafter in free form or in pharmaceutically acceptable salt form in the manufacture of a pharmaceutical composition for the treatment of allergic rhinitis, allergic dermatitis, drug allergy or food allergy, angioedema, urticaria, sudden infant death syndrome, bronchopulmonary aspergillosis, multiple sclerosis or mastocytosis; and to a method of treatment of warmblooded animals, preferably humans, in which a therapeutically effective dose of a compound of formula I is administered to a warm-blooded animal suffering from one of the diseases mentioned above.
• the present invention relates the use of N-phenyl-2-pyrimidine-amine derivatives of formula I
• R. is 4-pyrazinyl; 1 -methyl- 1 H-pyrrolyl; amino- or amino-lower alkyl-substituted phenyl, wherein the amino group in each case is free, alkylated or acylated; 1H-indolyl or 1 H- imidazolyl bonded at a five-membered ring carbon atom; or unsubstituted or lower alkyl- substituted pyridyl bonded at a ring carbon atom and unsubstituted or substituted at the nitrogen atom by oxygen;
• R 2 and R 3 are each independently of the other hydrogen or lower alkyl; one or two of the radicals Fit, R 5 , R 6 , R and R 8 are each nitro, fluoro-substituted lower alkoxy or a radical of formula II
• R 9 is hydrogen or lower alkyl
• X is oxo, thio, imino, N-lower alkyl-imino, hydroximino or O-lower alkyl-hydroximino,
• Y is oxygen or the group NH, n is 0 or 1 and
• R 10 is an aliphatic radical having at least 5 carbon atoms, or an aromatic, aromatic-aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, heterocyclic or heterocyclic-aliphatic radical, and the remaining radicals R 4 , R 5 , R 6 , R 7 and R 8 are each independently of the others hydrogen, lower alkyl that is unsubstituted or substituted by free or alkylated amino, piperazinyl, piperidinyl, pyrrolidinyl or by morpholinyl, or lower alkanoyl, trifluoromethyl, free, etherified or esterifed hydroxy, free, alkylated or acylated amino or free or estehfied carboxy,
• a medicament for treating allergic rhinitis, allergic dermatitis, drug allergy or food allergy, angioedema, urticaria, sudden infant death syndrome, bronchopulmonary aspergillosis, mastocytosis or multiple sclerosis for the manufacture of a medicament for treating allergic rhinitis, allergic dermatitis, drug allergy or food allergy, angioedema, urticaria, sudden infant death syndrome, bronchopulmonary aspergillosis, mastocytosis or multiple sclerosis.
• 1-methyl-1 H-pyrrolyl is preferably 1-methyl-1 H-pyrrol-2-yl or 1-methyl-1 H-pyrrol-3-yl.
• Amino- or amino-lower alkyl-substituted phenyl Ri wherein the amino group in each case is free, alkylated or acylated is phenyl substituted in any desired position (ortho, meta or para) wherein an alkylated amino group is preferably mono- or di-lower alkylamino, for example, dimethylamino, and the lower alkyl moiety of amino-lower alkyl is preferably linear C ⁇ -C 3 alkyl, such as especially methyl or ethyl.
• 1 H-indolyl bonded at a carbon atom of the five-membered ring is 1 H-indol-2-yl or 1 H-indol-3- yi.
• Unsubstituted or lower alkyl-substituted pyridyl bonded at a ring carbon atom is lower alkyl- substituted or preferably unsubstituted 2-, 4- or preferably 3-pyridyl, for example 3-pyridyl, 2-methyl-3-pyridyl or 4-methyl-3-pyridyl.
• Pyridyl substituted at the nitrogen atom by oxygen is a radical derived from pyridine N-oxide, i.e., N-oxido-pyridyl.
• Fluoro-substituted lower alkoxy is lower alkoxy carrying at least one, but preferably several, fluoro substituents, especially trifluoromethoxy or 1 ,1 ,2,2-tetrafluoro-ethoxy.
• X is oxo, thio, imino, N-lower alkyl-imino, hydroximino or O-lower alkyl-hydroximino
• X is preferably oxo.
• n is preferably 0, i.e. the group Y is not present.
• Y if present, is preferably the group NH.
• Lower alkyl R ⁇ R 2 , R 3 and R 9 is preferably methyl or ethyl.
• An aliphatic radical R 10 having at least 5 carbon atoms preferably has not more than 22 carbon atoms, generally not more than 10 carbon atoms, and is such a substituted or preferably unsubstituted aliphatic hydrocarbon radical, that is to say such a substituted or preferably unsubstituted alkynyl, alkenyl or preferably alkyl radical, such as C 5 -C 7 alkyl, for example n-pentyl.
• An aromatic radical R ⁇ 0 has up to 20 carbon atoms and is unsubstituted or substituted, for example, in each case unsubstituted or substituted naphthyl, such as especially 2-naphthyl, or preferably phenyl, the substituents preferably being selected from cyano, unsubstituted or hydroxy-, amino- or 4-methyl-piperazinyl-substituted lower alkyl, such as especially methyl, thfluoromethyl, free, etherified or esterified hydroxy, free, alkylated or acylated amino and free or esterified carboxy.
• an aromatic-aliphatic radical R 10 the aromatic moiety is as defined above and the aliphatic moiety is preferably lower alkyl, such as especially C C 2 alkyl, which is substituted or preferably unsubstituted, for example benzyl.
• a cycloaliphatic radical R ⁇ 0 has especially up to 30, more especially up to 20, and most especially up to 10 carbon atoms, is mono- or poly-cyclic and is substituted or preferably unsubstituted, for example, such a cycloalkyl radical, especially such a 5- or 6-membered cycloalkyl radical, such as preferably cyclohexyl.
• a cycloaliphatic-aliphatic radical R 10 the cycloaliphatic moiety is as defined above and the aliphatic moiety is preferably lower alkyl, such as especially d-C 2 alkyl, which is substituted or preferably unsubstituted.
• a heterocyclic radical R 10 contains especially up to 20 carbon atoms and is preferably a saturated or unsaturated monocyclic radical having 5- or 6-ring members and 1-3 hetero atoms which are preferably selected from nitrogen, oxygen and sulfur, especially, for example, thienyl or 2-, 3- or 4-pyridyl, or a bi- or tri-cyclic radical wherein, for example, one or two benzene radicals are annellated (fused) to the mentioned monocyclic radical.
• heterocyclic-aliphatic radical R 10 the heterocyclic moiety is as defined above and the aliphatic moiety is preferably lower alkyl, such as especially C 1 -C 2 alkyl, which is substituted or preferably unsubstituted.
• Etherified hydroxy is preferably lower alkoxy.
• Esterified hydroxy is preferably hydroxy esterified by an organic carboxylic acid, such as a lower alkanoic acid, or a mineral acid, such as a hydrohalic acid, for example, lower alkanoyloxy or especially halogen, such as iodine, bromine or especially fluorine or chlorine.
• Alkylated amino is, for example, lower alkylamino, such as methylamino, or di-lower alkylamino, such as dimethylamino.
• Acylated amino is, for example, lower alkanoylamino or benzoylamino.
• Esterified carboxy is, for example, lower alkoxycarbonyl, such as methoxycarbonyl.
• a substituted phenyl radical may carry up to 5 substituents, such as fluorine, but especially in the case of relatively large substituents is generally substituted by only from 1-3 substituents.
• substituents such as fluorine
• Examples of substituted phenyl that may be given special mention are 4-chloro-phenyl, pentafluoro-phenyl, 2-carboxy-phenyl, 2-methoxy-phenyl, 4-fluoro-phenyl, 4-cyano-phenyl and 4-methyl-phenyl.
• Salt-forming groups in a compound of formula I are groups or radicals having basic or acidic properties.
• Compounds having at least one basic group or at least one basic radical may form acid addition salts, for example, with inorganic acids, such as hydrochloric acid, sulfuric acid or a phosphoric acid, or with suitable organic carboxylic or sulfonic acids, for example, aliphatic mono- or di-carboxylic acids, such as trifluoroacetic acid, acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, fumaric acid, hydroxymaleic acid, malic acid, tartaric acid, citric acid or oxalic acid, or amino acids such as arginine or lysine, aromatic carboxylic acids, such as benzoic acid, 2-phenoxy-benzoic acid, 2-acetoxy-benzoic acid, salicylic acid, 4-aminos
• Compounds of formula I having acidic groups may form metal or ammonium salts, such as alkali metal or alkaline earth metal salts, for example, sodium, potassium, magnesium or calcium salts, or ammonium salts with ammonia or suitable organic amines, such as tertiary monoamines, for example, triethylamine or tri-(2-hydroxyethyl)-amine, or heterocyclic bases, for example, N-ethyl- piperidine or N,N'-dimethyl-piperazine.
• metal or ammonium salts such as alkali metal or alkaline earth metal salts, for example, sodium, potassium, magnesium or calcium salts, or ammonium salts with ammonia or suitable organic amines, such as tertiary monoamines, for example, triethylamine or tri-(2-hydroxyethyl)-amine, or heterocyclic bases, for example, N-ethyl- piperidine or N,N'-dimethyl-piperazine.
• the compounds of formula I can be used for the treatment or for the manufacture of medicaments for the treatment of warm-blooded animals, preferably humans.
• allergic rhinitis as used herein means any allergic reaction of the nasal mucosa. Such allergic reaction may occur, e.g., perennially, e.g., vernal conjunctivitis, or seasonally, e.g., hay fever.
• allergic dermatitis means especially atopic dermatitis, allergic contact dermatitis and eczematous dermatitis, but comprises, e.g., also seborrhoeic dermatitis, Lichen planus, urticaria and acne.
• Atopic dermatitis as defined herein is a chronic inflammatory skin disorder seen in individuals with a hereditary predisposition to a lowered cutaneous threshold to pruritus. It is principally characterized by extreme itching, leading to scratching and rubbing that in turns results in the typical lesions of eczema.
• Allergic contact dermatitis as defined herein is a form of dermatitis that is due to the allergic sensitization to various substances that produce inflammatory reactions in the skin of those who have acquired hypersensitivity to the allergen as a result of previous exposure to it.
• drug allergy or food allergy pertains to an allergic reaction produced by a drug or ingested antigens, such as, for example, strawberries, milk or eggs.
• bronchopulmonary aspergillosis relates to an infection of the lungs with Aspergillus.
• mastocytosis as used herein, relates to systemic mastocytosis, for example, mastocytoma, and particularly to canine mast cell neoplasms.
• mast cells play an important role as the primary effector cells in the allergic disorders mentioned herein. Antigen-specific IgE-mediated degranulation of mast cells leads to the subsequent release of chemical mediators and multiple cytokines and to leukotriene synthesis. Furthermore, mast cells are involved in the pathogenesis of multiple sclerosis.
• mast cell neoplasms occur in both humans and animals. In dogs, mast cell neoplasms are called mastocytomas, and the disease is common, representing 7-21% of canine tumors.
• human mastocytosis which is usually transient or indolent
• canine mast cell neoplasia which behaves unpredictably and is often aggressive and metastatic.
• human solitary mastocytomas essentially never metastasize; in contrast, 50% of canine mastocytomas behave in a malignant fashion, as estimated by Hottendorf & Nielsen (1969) after review of 46 published reports of tumors in 938 dogs.
• Cancer in the pet population is a spontaneous disease. Pet owners, motivated by prolonging the quality of their animals' life, frequently seek out the specialized care and treatment of veterinary oncologists at private referral veterinary hospitals and veterinary teaching hospitals across the country. Therapeutic modalities of veterinary cancer patients are similar to humans, including surgery, chemotherapy, radiation therapy, and biotherapy. It has been estimated that there are 42 million dogs and approximately 20 million cats in the United States. Using crude estimates of cancer incidence, there are roughly 4 million new cancer diagnoses made in dogs and a similar number in cats made each year.
• Cutaneous mast cell tumors in dogs are a common problem. Most mast cell tumors are benign and are cured with simple resection; however, if recurrent or metastatic to distant sites therapeutic options are limited. Treatment options for recurrent lesions can include external beam radiation therapy. For distant metastases or disseminated disease the use of Lomustine® and vinblastine containing chemotherapy protocols have demonstrated some benefit. Sites for metastases for mast cell tumors include skin, regional lymph nodes, spleen, liver and bone marrow.
• the KIT receptor's involvement in the pathogenesis of mastocytosis is suggested by the observation that several mutations resulting in constitutive activation of KIT have been detected in a number of mast cell lines. For instance, a point mutation in human c-KIT, causing substitution of Val for Asp816 in the phosphotransferase domain and receptor autoactivation, occurs in a long-term human mast cell leukemia line (HMC-1 ) and in the corresponding codon in two rodent mast cell lines. Moreover, this activating mutation has been identified in situ in some cases of human mastocytosis.
• HMC-1 human mast cell leukemia line
• the COMPOUNDS OF THE INVENTION are solubilized in PBS at a concentration of 10 "2 M and stored at -80°C.
• M-trans retinoic acid (Sigma) is dissolved in ethanol at a concentration of 10 "2 M, and stored in light-protected vials at -80°C.
• Purified mAb for tryptase (MAB1222) can be purchased from Chemicon International Inc., CA.
• the mAbs for CD34 (8G12, FITC) and CD11b (Leu15, PE) are purchased from Becton Dickinson Immunocytometry Systems (Mountain View, CA), and the mAb for CD41 (SZ22, FITC) from Immunotech S.A. (Marseilles, France).
• the mAb for glycophorin A (GPA, JC159, FITC) can be obtained from Dako (Glostrup, Denmark).
• the mAbs for c-kit (NU-c-kit) and for phosphotyrosine (4G10) can be purchased from Nichirei and Upstate Biotechnology, Inc (Lake Placid, NY), respectively.
• Serum-deprived liquid cultures are carried out in 24-well culture plates (#3047; Becton Dickinson). Twenty thousand CD34 + cells are cultured in each well containing 2 mL of ⁇ -medium supplemented with 1% BSA, 300 ⁇ g/mL fully iron-saturated human transferrin (approximately 98% pure, Sigma), 16 ⁇ g/mL soybean lecithin (Sigma), 9.6 ⁇ g/mL cholesterol (Nakalai Chemicals Ltd., Japan) and 20 ng/mL of SCF, 10 ng/mL of GM-CSF, 2 U/mL of EPO, 10 ng/mL of TPO, different concentrations of a COMPOUND OF THE INVENTION, alone or in combination.
• 10-week cultured cells grown with 20 ng/mL of SCF from CD34 + cord blood cells are used as target cells.
• Five to ten x 10 4 10-week cultured cells are incubated for 2 weeks in 24-well culture plates containing 20 ng/mL of SCF with or without various concentrations of a COMPOUND OF THE INVENTION.
• the plates are incubated at 37°C in a humidified atmosphere flushed with a mixture of 5% CO 2 , 5% O 2 and 90% N 2 .
• half of the culture medium is replaced every 2 weeks with fresh medium containing the factor(s).
• the number of viable cells is determined by a trypan-blue exclusion test using a hemocytometer.
• the mast cell colony assay is carried out in 35 mm Lux suspension culture dishes (#171099; Nunc, IL).
• the culture consisted of 10-week cultured cells (4,000 cells/mL) grown with 10 ng/mL of SCF, ⁇ -medium, 0.9% methylcellulose (Shinetsu Chemical, Japan), 1% BSA, 300 ⁇ g/mL of fully iron-saturated human transferrin, 16 ⁇ g/mL of soybean lecithin, 9.6 ⁇ g/mL of cholesterol and 100 ng/mL of SCF with or without 10 "6 M of a COMPOUND OF THE INVENTION.
• Dishes are incubated at 37°C in a humidified atmosphere flushed with a mixture of 5% CO 2 , 5% O 2 and 90% N 2 . After 4 weeks, aggregates consisting of 30 or more cells are scored as mast cell colonies, and those consisting of 10-29 cells as mast cell clusters. Thirty individual colonies and clusters are lifted, and stained with the anti-tryptase mAb or mouse lgG1 using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique. Almost all of the constituent cells are positive for tryptase.
• APAAP alkaline phosphatase-anti-alkaline phosphatase
• the cultured cells are spread on glass slides using a Cytospin II. Cytochemical reaction with peroxidase (POX) is performed by the conventional method. Reaction with mAb against tryptase is detected using the APAAP method (Dako APAAP Kit System, Dako Corp., CA), as described by Ma et al., Br. J. Haematol., Vol. 100, pp. 427-435 (1998).
• POX peroxidase
• FITC- or PE-mAb for the analysis of surface markers on the cultured cells, 1-2 x 10 5 cells are collected in plastic tubes and incubated with appropriately diluted FITC- or PE-mAb, as described by Kinoshita et al., Blood, Vol. 94, pp. 496-508 (1999). The cells are washed twice, after which their surface markers are analyzed with the FACScan flow cytometer. Viable cells are gated according to their forward light scatter characteristics and side scatter characteristics. The proportion of positive cells is determined by comparison to cells stained with FITC- or PE-conjugated mouse isotype-matched Ig. Detection of cellular apoptosis
• PI propidium iodide
• PI propidium iodide
• a percoll gradient centrifugation can be utilized. Ten-week cultured cells are layered on 27% Percoll (Sigma) in ⁇ -medium and 54% Percoll in PBS. After centrifugation, the cells are collected from the interface of the two different concentrations of Percoll solution, washed with PBS and treated with 1 mL of Ortho PermeaFixTM for 40 minutes at room temperature. The cells are then incubated with DNase-free RNase (Sigma) for 15 minutes at 37°C, and stained with PI for 15 minutes. The DNA content of 2 x 10 4 cells is monitored with a flow cytometer.
• the 10-week cultured cells (2 x 10 6 ) exposed to SCF or SCF and a COMPOUND OF THE INVENTION are lysed for 10 minutes on ice in 100 ⁇ L hypotonic lysis buffer [10 mM Tris (pH 7.5), 10 mM EDTA, pH 8.0, 0.5% Triton X-100]. After centrifugation for 10 minutes at 14,000 g, the supernatant is transferred to a new tube, and treated with 0.2 mg/mL RNase A (Sigma) and 0.2 mg/mL Proteinase K (Sigma). DNA is precipitated with 120 ⁇ L isopropanol and 20 ⁇ L 5 M NaCI overnight at -20°C. After centrifugation at 14,000 g, the pellets are dried, dissolved in 20 ⁇ L Tris-EDTA, and then samples are analyzed by gel electrophoresis in 2% agarose and ethidium bromide staining.
• hypotonic lysis buffer 10 mM Tris (p
• Histamine concentrations in cell lysates obtained by the treatment of the cultured cells with 0.5% Nonidet P-40 and in supernatant are measured by Histamine Radioimmunoassay (RIA) Kit (Immunotech), as described in Kinoshita et al., Blood, Vol. 94, pp. 496-508 (1999).
• STI571 B In SCF plus STI571 B, the number of viable cells is unchanged on day 2 and then declines in a time-dependent fashion. On day 14, the total cell number reaches to a negligible level. Consistent with the results obtained by 10-week cultured mast cells, STI571 B at 10 "6 M markedly depresses the cell production by CD34 + cells under stimulation with SCF. Furthermore, STI571 B induces a programmed death of the cultured mast cells grown under stimulation with SCF. The percentage of sub-diploid nuclei increases with the culture period. This observation can be confirmed by DNA laddering in cells exposed to SCF plus STI571 B on gel electrophoresis. STI571 B exerts its inhibitory effects at the early phase of mast cell development as well as the growth of the late-appearing mast cells.
• Antibodies A polyclonal rabbit anti-KIT antibody (c-kit Ab-1) was used at a dilution of 1 :500 (c-kit Ab-1 ; Oncogene, Cambridge, MA). An anti-phosphotyrosine antibody (PY20) was used at a dilution of 1:1000 (PY20 Transduction Laboratories; Lexington, KY). Peroxidase conjugated goat anti-mouse antibody was used at a dilution of 1 :5000 and goat anti-rabbit antibody at a dilution of 1 :10,000 (Pierce; Rockford, IL).
• Both cell lines were maintained in DMEM supplemented with 2% bovine calf, 1 mM L-glutamine, 12.5 mM HEPES (pH 7.5), 0.25 mg/ml Histidine, 1 % Penicillin-Streptomycin and 1% fungizone.
• the BR and C2 cells were derived from canine mast cell tumors and were originally established in long-term culture after initial passaging in immunodeficient mice (DeVinney R et al., Am J Respir Cell Mol Biol 1990; 3(5):413-420; Lazarus SC et al., Am J Physiol 1986; 251(6 Pt 1):C935-C944).
• the BR cell line has a point mutation (T1752C) resulting in a Leucine to Proline substitution at amino acid 575 (juxtamembrane domain).
• the C2 cell line has an internal tandem duplication (ITD) of the KIT juxtamembrane region. The translation of this ITD results in reduplication of amino acid residues at the 3' end of exon 11 (London et al., Exp. Hematol., Vol. 27, No. 4, pp. 689-697 (1999); Ma et al., J. Invest. Dermatol., Vol. 112, No. 2, pp. 165-170 (1999)).
• Proliferation assays Cells were added to 96-well plates at a density of 40,000 cells/well in normal culture media and varying concentrations of SALT I. Proliferation was measured at 48-72 hours using an XTT-based assay (Roche Molecular Biochemicals; Indianapolis, IN) (Heinrich et al., Blood, Vol. 96, No. 3, pp. 925-932 (2000))
• Protein lysates BR and C2 cells were washed x 2 in PBS and then quiesced in Optimem (Gibco-BRL) at 37°C for approximately 18 hours. Cells were then incubated for 90 minutes in the presence of various concentrations of SALT I. Following this incubation, the cells were pelleted and lysed using 100-250 ⁇ L of protein lysis buffer (50 mM Tris, 150 mM NaCI, 1% NP-40, 0.25% Deoxycholate, with addition of the inhibitors aprotinin, leupeptin, pepstatin, PMSF and sodium orthovanadate [Sigma]). Western immunoblot analysis was performed as previously described (Hoatlin et al., Blood, Vol. 91 , No. 4, pp. 1418-1425 (1998); Heinrich et al., Blood, Vol. 96, No. 3, pp. 925-932 (2000)).
• Example 3 COMPOUND I inhibits the constitutively activated KIT kinase associated with canine mast cell tumors
• Lysates prepared from BR or C2 cells were probed with an anti-P-Tyr antibody and KIT receptor activation was assessed by measuring autophosphorylation. As reported previously, KIT autophosphorylation in these cells was observed in the absence of SLF (Ma et al., J. Invest. Dermatol., Vol. 112, No. 2, pp. 165-170 (1999); Ma et al., J. Invest. Dermatol., Vol. 114, No. 2, pp. 392-394 (2000)). Inhibition of KIT autophosphorylation by COMPOUND I was dose dependent with complete inhibition observed using 10 and 1.0 ⁇ M doses. Near complete inhibition was seen using a dose of 0.1 ⁇ M.
• COMPOUND I not only inhibits the autophosphorylation of the mutated c-kit receptor in these cells, but also is a more potent inhibitor of this mutated receptor than it is of the wild type c-kit receptor (IC 50 100-200 nM) (Heinrich et al., Blood, Vol. 96, No. 3, pp. 925-932 (2000)).
• IC 50 100-200 nM wild type c-kit receptor
• Example 4 COMPOUND I inhibits the proliferation of cell lines of canine mast cell tumors
• BR or C2 cells were plated in 96-well plates at a concentration of 40,000 cells/well and cultured in normal growth media and varying concentration of COMPOUND I.
• Cellular proliferation was measured at 72 hours using an XTT-based assay system.
• Each COMPOUND I concentration was assayed in triplicate. Results are expressed as a percent of maximal proliferation (cells only, no COMPOUND I) ⁇ 1 standard deviation. Representative results from one of six independent experiments are shown.
• compositions for pets and humans are provided.
• Example 5 Capsules with 4-[(4-methyl-1-piperazin-1-ylmethyl)-N-[4-methyl-3-[[4-(3- pyridinyl)-2-pyrimidinyl]amino]phenyl]benzamide methanesulfonate, ⁇ -crystal form
• the capsules are prepared by mixing the components and filling the mixture into hard gelatin capsules, size 1.
• Example 6 Capsules with 4-[(4-methyl-1-piperazin-1-ylmethyl)-N-[4-methyl-3-[[4-(3- pyridinyl)-2-pyrimidinyl]amino]phenyl]benzamide methanesulfonate, ⁇ -crystal form
• the capsules are prepared by mixing the components and filling the mixture into hard gelatin capsules, size 1.
• Example 7 Capsules with 4-[(4-methyl-1-piperazin-1-ylmethyl)-N-[4-methyl-3-[[4-(3- pyridinyl)-2-pyrimidinyl]amino]phenyl]benzamide methanesulfonate, ⁇ -crystal form.
• the capsules are prepared by mixing the components and filling the mixture into hard gelatin capsules, size 1.
• Example 8 Example of a prospective case series of pet dogs with measurable cutaneous mast cell tumors.
• the study patients are pet dogs with measurable and histologically-confirmed mast cell tumors. Cases are limited to those with measurable lesions amenable to biopsy.
• Pretreatment evaluation of all cases include physical examination, complete blood count, buffy coat, serum biochemistry, urinalysis, serum bile acids (fasting and post-prandial), documentation of regional lymph node size, abdominal radiographs, and abdominal ultrasound.
• the treatment regimen is 25 mg/kg PO QD x 60 days of SALT I.
• Treatment is continued in all cases for 60 days unless disease progression is noted. In cases experiencing partial response or complete response ongoing therapy for an additional 60 days may be considered. Cases successfully completing therapy are eligible for repeat entry to study.
• initial staging consists of physical examination, CBC, buffy coat, serum biochemistry, liver function tests (serum bile acids), urinalysis, abdominal radiographs, and abdominal ultrasound.
• Re-evaluation of may consist of physical examination and measurement of tumor burden alone or repeat clinical staging.
• Tumor burden is measured at days 0, 7, 14 and 28, and then every 14 days. Treatment response will be defined against measurable cutaneous lesion(s) and other lesions identified at staging (CR, PR, SD, PD - defined below). Collection of plasma from the first 5 entered cases is undertaken at 0, 0.5, 1 , 2, 5, 8, 12, 16 and 24 hours following first dose of SALT I.
• COMPOUND I is assessed against measurable cutaneous mast cell tumors, using clinical endpoints.
• Biological endpoints may be taken from serial biopsies collected from cutaneous tumors and from blood samples available through the treatment course.
• Clinical endpoints include response rate of measurable tumors, objective response against measurable tumor, and time to progression of measurable tumor. All adverse side effects will be recorded.
• CR Complete Responses
• PR Partial Responses
• SD Stable Disease
• PD Progressive Disease
• R Relapse of disease
• TTP Time To Progression
• Duration of Remission and Survival may increase in animals under treatment with COMPOUND I.
• CR is defined as disappearance of all clinical evidence of cancer and of any signs related to the cancer.
• PR is defined as a 50% or greater decrease in the sum of the products of measurements for representative lesions, without an increase in size of any lesions or appearance of any new lesions.
• SD is defined as no response or a response of less than that defined for partial response or progressive disease without appearance of any new lesions or worsening of clinical signs.
• PD is defined as an unequivocal increase of at least 50% in the size of any measurable lesion or appearance of new lesions.
• R is defined as appearance of new lesions or reappearance of old lesions in dogs that had had a CR; in dogs that had had only a PR, R was defined as at least a 50% increase in the sum of the products of measurements of representative lesions, compared with measurements obtained at the time of maximum response.
• TTP is reported from day 0 of the protocol. TTP will be defined as the number of days start of therapy (from day 0) to R.
• Duration of Remission is defined as the number of days from the objective response (PR or CR) to R.
• “Survival” is defined as the number of days from the start of treatment with COMPOUND I to death. Cause of death will be noted but may include disease progression, toxicity and other.
• radicals F , R 5 , R 6 , R 7 and R 8 are each nitro or a radical of formula II wherein R 9 is hydrogen or lower alkyl, X is oxo, thio, imino, N-lower alkyl-imino, hydroximino or O-lower alkyl-hydroximino, Y is oxygen or the group NH, n is 0 or 1 and Rio is an aliphatic radical having at least 5 carbon atoms or an aromatic, aromatic- aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, heterocydic or heterocyclic-aliphatic radical, and the remaining radicals R-s, R 5 , R 6 , R and R 8 are each independently of the others hydrogen, lower alkyl that is unsubstituted or substituted by free or alkylated amino, piperazinyl, piperidinyl, pyrroli
• R T is pyridyl or N-oxido-pyridyl each of which is bonded at a carbon atom
• R 2 and R 3 are each hydrogen
• P is hydrogen or lower alkyl
• R 5 is hydrogen, lower alkyl or trifluoromethyl
• R 6 is hydrogen
• R 7 is nitro, fluoro-substituted lower alkoxy or a radical of formula II wherein
• R 9 is hydrogen, X is oxo, n is 0 and R 1 0 is pyridyl bonded at a carbon atom, phenyl that is unsubstituted or substituted by halogen, cyano, lower alkoxy, carboxy, lower alkyl or by 4-methyl-piperazinyl-methyl, or C 5 -C 7 alkyl, thienyl, 2-naphthyl or cyclohexyl, and
• R 8 is hydrogen
• Ri is pyridyl bonded at a carbon atom
• R 3 , R5, Re and R 8 are each hydrogen
• R 4 is lower alkyl
• R 7 a radical of formula II wherein R 9 is hydrogen, X is oxo, n is 0 and R 10 is 4-methyl- piperazinyl-methyl.
• the disease to be treated is selected from allergic rhinitis, allergic dermatitis, drug allergy and food allergy.
• the disease to be treated is multiple sclerosis.
• the disease to be treated is selected from angioedema, urticaria, sudden infant death syndrome and bronchopulmonary aspergillosis.
• the COMPOUNDS OF THE INVENTION can be used for the treatment of systemic mastocytosis, especially mastocytoma. The latter disease is a malignant disease with extensive metastasis, in particular in dogs.
• the COMPOUNDS OF THE INVENTION are particularly useful for the preparation of a medicament for treating canine mast cell neoplasms.
• COMPOUNDS OF THE INVENTION are generically and specifically disclosed in the patent applications EP 0 564 409 A1 and WO 99/03854, in particular in the compound claims and the final products of the working examples, the subject-matter of the final products, the pharmaceutical preparations and the claims are hereby incorporated into the present application by reference to these publications. Comprised are likewise the corresponding stereoisomers as well as the corresponding polymorphs, e.g., crystal modifications, which are disclosed therein.
• COMPOUNDS OF THE INVENTION are described to be useful for the therapy of cancer, thrombosis, psoriasis, fibrosis, dermatosclerosis and atheriosclerosis.
• COMPOUNDS OF THE INVENTION surprisingly have a beneficial effect on allergic rhinitis, allergic dermatitis, drug allergy or food allergy, angioedema, urticaria, sudden infant death syndrome, bronchopulmonary aspergillosis, multiple sclerosis, and, moreover, mastocytosis, especially canine mast cell neoplasms.
• the present invention also provides a method of treatment of warm-blooded animals, including humans, in which an therapeutically effective dose of a COMPOUND OF THE INVENTION is administered to such a warm-blooded animal, preferably a human or a dog, very preferably a human, suffering from one of the diseases mentioned herein.
• the invention relates also to a method for administering to a dog subject having canine mast cell neoplasms a COMPOUND OF THE INVENTION or a pharmaceutically acceptable salt thereof, which comprises administering a pharmaceutically effective amount of a COMPOUND OF THE INVENTION or a pharmaceutically acceptable salt thereof to the dog once daily for preferably a period exceeding 1 , 2 or even 3 months.
• the invention relates especially to such method wherein a daily dose of about 20-200 mg, preferably 80-160 mg, especially 125 mg of SALT I is administered.
• the invention relates to a use or method wherein a daily dose of a monomethanesulfonate salt of 4-(4-methylpiperazin-1-ylmethyl)-N-[4-methyl-3-(4-pyridin-3- yl)pyrimidin-2-ylamino)phenyl]-benzamide of the formula I is administered to a dog, and said daily dose comprises an amount of said monomethanesulfonate salt sufficient to maintain plasma levels of at least 0.2 ⁇ M.
• method of treatment as used herein relates especially also to a method of prevention of the diseases mentioned herein, i.e., the prophylactic administration of a pharmaceutical composition comprising a COMPOUND OF THE INVENTION to healthy patients to prevent the outbreak of the diseases mentioned herein.
• the present invention relates also to a pharmaceutical composition for the treatment of at least one of the diseases mentioned herein comprising a COMPOUND OF THE INVENTION.
• Pharmaceutical compositions for the treatment of at least one of the diseases mentioned herein comprise an effective amount of the COMPOUNDS OF THE INVENTION together with pharmaceutically acceptable carriers that are suitable for topical, enteral, for example oral or rectal, or parenteral administration, and may be inorganic or organic, solid or liquid.
• tablets or gelatin capsules comprising the COMPOUNDS OF THE INVENTION together with diluents, and/or lubricants, for example, silicic acid, talc, stearic acid or salts thereof, and/or polyethylene glycol, but also lotions, gels or creams.
• Tablets may comprise binders, starches, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, disintegrators, and/or effervescent mixtures, or adsorbents, dyes, flavourings and sweeteners.
• the COMPOUNDS OF THE INVENTION can also be used in the form of parenterally administrable compositions or in the form of infusion solutions.
• Topical administration is, e.g., to the skin.
• a further form of topical administration is to the eye, e.g., for the treatment of vernal conjunctivitis.
• the pharmaceutical compositions may be sterilised and/or may comprise excipients, for example, preservatives, stabilisers, wetting agents and/or emulsifiers, solubilisers, salts for regulating the osmotic pressure and/or buffers.
• the present pharmaceutical compositions are prepared in a manner known per se, for example, by means of conventional mixing, granulating, confectioning, dissolving or lyophilising processes, and comprise approximately from 1-100%, especially from approximately 1% to approximately 20%, active ingredient(s).
• the COMPOUNDS OF THE INVENTION can, for example, be formulated as disclosed in Examples 4 and 6 of WO 99/03854.
• the dosage range of the COMPOUNDS OF THE INVENTION to be employed depends upon factors known to the person skilled in the art including species of the warm-blooded animal, body weight and age, the mode of administration, the particular substance to be employed and the disease to be treated. Unless stated otherwise herein, the COMPOUNDS OF THE INVENTION are preferably administered from one to four times per day or immediately when an allergic reaction is observed.
• the COMPOUNDS OF THE INVENTION are preferably administered to a warm-blooded animal, especially a human in a dosage in the range of about 10-750 mg/day, preferably 30-600 mg/day more preferably 30-300 mg/day.
• effective doses for example, daily doses of about 20-200 mg, preferably 80-160 mg, especially 125 mg, are administered to warm-blooded animals of about 5 kg bodyweight.
• a starting dose of 125 mg daily can be recommended.
• dose escalation can be safely considered and dogs may be treated as long as they benefit from treatment and in the absence of limiting toxicities.
• Dosages may be titered so as to achieve plasma levels of at least 0.2 ⁇ M (micromolar), preferably at least 0.5 ⁇ M, more preferably at least 1 ⁇ M. Achieving and/or maintaining a plasma level of about 1 ⁇ M is particularly preferred.

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• Health & Medical Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Chemical & Material Sciences (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Medicinal Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Life Sciences & Earth Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Organic Chemistry (AREA)
• General Chemical & Material Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Epidemiology (AREA)
• Pulmonology (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Immunology (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Pyridine Compounds (AREA)

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• 2001
  • 2001-04-05 GB GBGB0108606.5A patent/GB0108606D0/en not_active Ceased
• 2002
  • 2002-04-05 KR KR10-2003-7012939A patent/KR20040007485A/ko not_active Application Discontinuation
  • 2002-04-05 EP EP02763956A patent/EP1389113A1/en not_active Ceased
  • 2002-04-05 SK SK1231-2003A patent/SK12312003A3/sk not_active Application Discontinuation
  • 2002-04-05 BR BR0208663-8A patent/BR0208663A/pt not_active IP Right Cessation
  • 2002-04-05 CA CA002443092A patent/CA2443092A1/en not_active Abandoned
  • 2002-04-05 PL PL02363080A patent/PL363080A1/xx unknown
  • 2002-04-05 CZ CZ20032705A patent/CZ20032705A3/cs unknown
  • 2002-04-05 NZ NZ528934A patent/NZ528934A/en not_active IP Right Cessation
  • 2002-04-05 HU HU0303751A patent/HUP0303751A3/hu unknown
  • 2002-04-05 AU AU2002307140A patent/AU2002307140B2/en not_active Expired
  • 2002-04-05 KR KR1020097003644A patent/KR20090034998A/ko not_active Application Discontinuation
  • 2002-04-05 MX MXPA03009058A patent/MXPA03009058A/es not_active Application Discontinuation
  • 2002-04-05 WO PCT/US2002/010742 patent/WO2002080925A1/en active Application Filing
  • 2002-04-05 CN CNA028089707A patent/CN1529602A/zh active Pending
  • 2002-04-05 IL IL15806102A patent/IL158061A0/xx unknown
  • 2002-04-05 RU RU2003130635/15A patent/RU2304436C2/ru not_active IP Right Cessation
• 2003
  • 2003-09-29 ZA ZA200307563A patent/ZA200307563B/en unknown
  • 2003-10-02 NO NO20034414A patent/NO325454B1/no not_active IP Right Cessation

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