ES2385443A1 - Pyrimidine urea-derived compound for treating inflammatory diseases - Google Patents
Pyrimidine urea-derived compound for treating inflammatory diseases Download PDFInfo
- Publication number
- ES2385443A1 ES2385443A1 ES201031978A ES201031978A ES2385443A1 ES 2385443 A1 ES2385443 A1 ES 2385443A1 ES 201031978 A ES201031978 A ES 201031978A ES 201031978 A ES201031978 A ES 201031978A ES 2385443 A1 ES2385443 A1 ES 2385443A1
- Authority
- ES
- Spain
- Prior art keywords
- substituted
- unsubstituted
- clo
- pharmaceutical composition
- use according
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 72
- 208000027866 inflammatory disease Diseases 0.000 title claims abstract description 31
- ZENDWEPAVHORFD-UHFFFAOYSA-N pyrimidine;urea Chemical compound NC(N)=O.C1=CN=CN=C1 ZENDWEPAVHORFD-UHFFFAOYSA-N 0.000 title claims abstract description 8
- 206010040047 Sepsis Diseases 0.000 claims abstract description 21
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 19
- 241000894006 Bacteria Species 0.000 claims abstract description 16
- 230000001154 acute effect Effects 0.000 claims abstract description 15
- 206010040070 Septic Shock Diseases 0.000 claims abstract description 13
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 claims abstract description 7
- 208000019693 Lung disease Diseases 0.000 claims abstract description 7
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 claims abstract description 7
- 231100000354 acute hepatitis Toxicity 0.000 claims abstract description 7
- 206010003246 arthritis Diseases 0.000 claims abstract description 7
- 230000004064 dysfunction Effects 0.000 claims abstract description 7
- 208000006454 hepatitis Diseases 0.000 claims abstract description 7
- 230000002107 myocardial effect Effects 0.000 claims abstract description 7
- 208000021043 septic peritonitis Diseases 0.000 claims abstract description 7
- 230000036303 septic shock Effects 0.000 claims abstract description 7
- -1 alkenyl ether Chemical compound 0.000 claims description 17
- 239000003814 drug Substances 0.000 claims description 14
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- 125000000623 heterocyclic group Chemical group 0.000 claims description 12
- 125000004432 carbon atom Chemical group C* 0.000 claims description 8
- UHOVQNZJYSORNB-UHFFFAOYSA-N monobenzene Natural products C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 8
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 8
- 208000037487 Endotoxemia Diseases 0.000 claims description 6
- 206010014824 Endotoxic shock Diseases 0.000 claims description 6
- 201000000028 adult respiratory distress syndrome Diseases 0.000 claims description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- 125000004446 heteroarylalkyl group Chemical group 0.000 claims description 6
- 230000002956 necrotizing effect Effects 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- 206010023424 Kidney infection Diseases 0.000 claims description 4
- 206010037596 Pyelonephritis Diseases 0.000 claims description 4
- 125000004429 atom Chemical group 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 229940124597 therapeutic agent Drugs 0.000 claims description 4
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 3
- 125000003118 aryl group Chemical group 0.000 claims description 3
- 125000000392 cycloalkenyl group Chemical group 0.000 claims 2
- 239000002131 composite material Substances 0.000 claims 1
- 125000000753 cycloalkyl group Chemical group 0.000 claims 1
- 208000015181 infectious disease Diseases 0.000 abstract description 4
- 231100000284 endotoxic Toxicity 0.000 abstract 2
- 230000002346 endotoxic effect Effects 0.000 abstract 2
- 206010051606 Necrotising colitis Diseases 0.000 abstract 1
- 208000004995 necrotizing enterocolitis Diseases 0.000 abstract 1
- 201000006195 perinatal necrotizing enterocolitis Diseases 0.000 abstract 1
- 230000035939 shock Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 37
- 239000002158 endotoxin Substances 0.000 description 35
- 229920006008 lipopolysaccharide Polymers 0.000 description 35
- 241000588724 Escherichia coli Species 0.000 description 18
- 241000894007 species Species 0.000 description 12
- 102000004127 Cytokines Human genes 0.000 description 8
- 108090000695 Cytokines Proteins 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 6
- 150000003672 ureas Chemical class 0.000 description 6
- 238000001262 western blot Methods 0.000 description 6
- 230000002538 fungal effect Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 230000000770 proinflammatory effect Effects 0.000 description 5
- 102000007469 Actins Human genes 0.000 description 4
- 108010085238 Actins Proteins 0.000 description 4
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 description 4
- 102000004890 Interleukin-8 Human genes 0.000 description 4
- 108090001007 Interleukin-8 Proteins 0.000 description 4
- 102100039360 Toll-like receptor 4 Human genes 0.000 description 4
- 229940122954 Transcription factor inhibitor Drugs 0.000 description 4
- 230000003110 anti-inflammatory effect Effects 0.000 description 4
- 230000000845 anti-microbial effect Effects 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 235000013877 carbamide Nutrition 0.000 description 4
- 108091006086 inhibitor proteins Proteins 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 3
- 239000002246 antineoplastic agent Substances 0.000 description 3
- 229940041181 antineoplastic drug Drugs 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000030833 cell death Effects 0.000 description 3
- 230000001472 cytotoxic effect Effects 0.000 description 3
- 230000009036 growth inhibition Effects 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 239000006180 TBST buffer Substances 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000007398 colorimetric assay Methods 0.000 description 2
- 238000002784 cytotoxicity assay Methods 0.000 description 2
- 231100000263 cytotoxicity test Toxicity 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000007877 drug screening Methods 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- AWJUIBRHMBBTKR-UHFFFAOYSA-N isoquinoline Chemical compound C1=NC=CC2=CC=CC=C21 AWJUIBRHMBBTKR-UHFFFAOYSA-N 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- WUGFKDNWMUDOPN-UHFFFAOYSA-N 2-(2H-pyrimidin-1-yl)acetohydrazide Chemical class NNC(=O)CN1CN=CC=C1 WUGFKDNWMUDOPN-UHFFFAOYSA-N 0.000 description 1
- DVLFYONBTKHTER-UHFFFAOYSA-N 3-(N-morpholino)propanesulfonic acid Chemical compound OS(=O)(=O)CCCN1CCOCC1 DVLFYONBTKHTER-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 125000000041 C6-C10 aryl group Chemical group 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 206010014733 Endometrial cancer Diseases 0.000 description 1
- 206010014759 Endometrial neoplasm Diseases 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical group N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 108010060804 Toll-Like Receptor 4 Proteins 0.000 description 1
- 102000008233 Toll-Like Receptor 4 Human genes 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- WWSLNWLSIWYAJL-UHFFFAOYSA-N acetyl azide Chemical class CC(=O)N=[N+]=[N-] WWSLNWLSIWYAJL-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 150000004657 carbamic acid derivatives Chemical class 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000011278 co-treatment Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- 229940126601 medicinal product Drugs 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 108020004084 membrane receptors Proteins 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 239000003656 tris buffered saline Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
- C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
- C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
- C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
- C07D239/52—Two oxygen atoms
- C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
-
- 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
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Rheumatology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pain & Pain Management (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
USO DE COMPUESTOS DERIVADOS DE UREA PIRIMIDíNICA PARA LA ELABORACiÓN DE UN MEDICAMENTO ÚTIL PARA EL TRATAMIENTO DE ENFERMEDADES INFLAMATORIAS USE OF COMPOUNDS DERIVED FROM PYRIMIDINE UREA FOR THE DEVELOPMENT OF A USEFUL MEDICINAL PRODUCT FOR THE TREATMENT OF INFLAMMATORY DISEASES
La presente invención se refiere a compuestos derivados de urea pirimidínica que tienen actividad anti-inflamatoria. Dichos compuestos son útiles para preparar composiciones farmacéuticas para el tratamiento de enfermedades inflamatorias, preferiblemente sepsis, artritis, infecciones renales, enfermedades pulmonares agudas inducidas por sepsis, síndrome de distrés respiratorio agudo inducido por sepsis, shock endotóxico o endotoxemia, peritonitis séptica, hepatitis aguda, disfunción miocárdica aguda inducida por shock séptico y enterecolitis necrotizante. The present invention relates to compounds derived from pyrimidine urea that have anti-inflammatory activity. Such compounds are useful for preparing pharmaceutical compositions for the treatment of inflammatory diseases, preferably sepsis, arthritis, kidney infections, acute pulmonary diseases induced by sepsis, acute respiratory distress syndrome induced by sepsis, endotoxic shock or endotoxemia, septic peritonitis, acute hepatitis, acute myocardial dysfunction induced by septic shock and necrotizing enterecolitis.
En la técnica existen diversos compuestos con capacidad para disminuir los efectos inflamatorios producidos por acción de los Lipopolisacáridos (LPS). Es el caso de la inosina, que reduce la toxicidad de las citoquinas en células pulmonares en experimentos in vitro y suprime la inflamación pulmonar inducida por LPS en experimentos in vivo. In the art there are several compounds with the capacity to reduce the inflammatory effects produced by the action of Lipopolysaccharides (LPS). This is the case of inosine, which reduces the toxicity of cytokines in lung cells in in vitro experiments and suppresses LPS-induced lung inflammation in in vivo experiments.
Existen varios documentos que relacionan el uso de ureas con una posible acción anti-inflamatoria. El documento WOOO/43384 describe ureas heterocíclicas aromáticas que inhiben la producción de citoquinas involucradas en los procesos inflamatorios. El documento US 2005/0239811 A1 explica el uso de ureas N-1, 1, 3 trisustituídas que inhiben la liberación de citoquinas por parte de los lipopolisacáridos. El documento US 2003/0216396 A1 describe el uso de compuestos ureas conteniendo una piridina, quinolina o isoquinolina y que funcionalmente serían de utilidad en el tratamiento de enfermedades inflamatorias como la osteoporosis o en los desórdenes patológicos relacionados con la angiogénesis. There are several documents that relate the use of ureas with a possible anti-inflammatory action. WOOO / 43384 describes aromatic heterocyclic ureas that inhibit the production of cytokines involved in inflammatory processes. US 2005/0239811 A1 explains the use of trisubstituted ureas N-1, 1, 3 that inhibit the release of cytokines by lipopolysaccharides. US 2003/0216396 A1 describes the use of urea compounds containing a pyridine, quinoline or isoquinoline and which would functionally be useful in the treatment of inflammatory diseases such as osteoporosis or in pathological disorders related to angiogenesis.
DESCRIPCiÓN DE LA INVENCiÓN DESCRIPTION OF THE INVENTION
La presente invención es un compuesto derivado de urea pirimidínica de fórmula (1): donde: The present invention is a compound derived from pyrimidine urea of formula (1): where:
Res Beef
o or
5 donde Ra está seleccionado entre el grupo formado por H, carboxilo, CF3, F, CI, Br, 1, éter de alquilo C1-ClO sustituido o no sustituido, éter de alquenilo C2-ClO sustituido o no sustituido, éter de alquinilo CrClO sustituido o no sustituido, alquilo C1-ClO sustituido o no sustituido, alquenilo Cr ClO sustituido 5 where Ra is selected from the group consisting of H, carboxyl, CF3, F, CI, Br, 1, substituted or unsubstituted C1-ClO alkyl ether, substituted or unsubstituted C2-ClO alkenyl ether, CrClO alkynyl ether substituted or unsubstituted, substituted or unsubstituted C1-ClO alkyl, substituted Cr ClO alkenyl
10 o no sustituido, alquinilo C2-ClO sustituido o no sustituido, cicloalquilo C3-ClO sustituido o no sustituido, cicloalquenilo C5-ClO sustituido o no sustituido, cicloalquinilo Ca-ClO sustituido o no sustituido, arilo C6-ClO sustituido o no sustituido, aralquilo Cr ClO sustituido o no sustituido, heterociclilo de 3-10 miembros de anillo sustituido o no sustituido y heteroarilalquilo sustituido o no 10 or unsubstituted, substituted or unsubstituted C2-ClO alkynyl, substituted or unsubstituted C3-ClO cycloalkyl, substituted or unsubstituted C5-ClO cycloalkenyl, substituted or unsubstituted Ca-ClO cycloalkynyl, substituted or unsubstituted C6-ClO aryl, aralkyl Cr ClO substituted or unsubstituted, 3-10 substituted or unsubstituted heterocyclyl ring and substituted or unsubstituted heteroarylalkyl
15 sustituido de 2 a 10 átomos de carbono, y ambos heterociclos de 1 a 3 átomos diferentes al carbono, para el tratamiento de enfermedades inflamatorias. 15 substituted from 2 to 10 carbon atoms, and both heterocycles of 1 to 3 atoms other than carbon, for the treatment of inflammatory diseases.
En la presente invención, se entiende. por "enfermedades inflamatorias" a In the present invention, it is understood. for "inflammatory diseases" to
20 enfermedades que cursan con inflamación. En este grupo se encuentran las infecciones causadas por bacterias que cursan con inflamación, como por ejemplo, la sepsis. También se encuentran dentro de este grupo la artritis, infecciones renales, enfermedades pulmonares agudas inducidas por sepsis, síndrome de distrés respiratorio agudo inducido por sepsis, shock endotóxico o endotoxemia, 20 diseases that occur with inflammation. In this group are infections caused by bacteria that occur with inflammation, such as sepsis. Also included in this group are arthritis, kidney infections, acute pulmonary diseases induced by sepsis, acute respiratory distress syndrome induced by sepsis, endotoxic shock or endotoxemia,
25 peritonitis séptica, hepatitis aguda, disfunción miocárdica aguda inducida por shock séptico y enterecolitis necrotizante. 25 septic peritonitis, acute hepatitis, acute myocardial dysfunction induced by septic shock and necrotizing enterecolitis.
Los compuestos derivados de urea pirimidínica descritos en la presente invención poseen actividad anti-inflamatoria ya que son capaces de disminuir los efectos antiproliferativos inducidos por LPS de bacterias Gram-negativas sobre la línea celular A549, inhibir el aumento de las citoquinas pro-inflamatorias IL-8 e IL-1P inducidas por The compounds derived from pyrimidine urea described in the present invention possess anti-inflammatory activity since they are capable of decreasing the anti-proliferative effects induced by LPS of Gram-negative bacteria on the A549 cell line, inhibiting the increase of IL-pro-inflammatory cytokines 8 and IL-1P induced by
5 LPS y de inhibir el aumento de expresión de la proteína receptora TLR4 y de inhibir la disminución de la proteína inhibidora del factor de transcripción NF-KB inducidos por LPS. 5 LPS and to inhibit the increase in expression of the TLR4 receptor protein and to inhibit the decrease in the NF-KB transcription factor inhibitor protein induced by LPS.
Una realización preferible es un compuesto de la invención, donde R es benceno A preferable embodiment is a compound of the invention, where R is benzene.
Otra realización de la invención es un compuesto de la invención, donde R es Another embodiment of the invention is a compound of the invention, where R is
Otra realización de la invención es un compuesto de la invención, donde Res Another embodiment of the invention is a compound of the invention, wherein Res
II
Otra realización de la invención es un compuesto de la invención, donde R es Another embodiment of the invention is a compound of the invention, where R is
)JF ) JF
20 Otra realización de la invención es un compuesto de la invención, donde Res Another embodiment of the invention is a compound of the invention, wherein Res
CI I~ ~ CI I ~ ~
el he
Otra realización de la invención es un compuesto de la invención, donde R es Another embodiment of the invention is a compound of the invention, where R is
F F
F F
Otra realización de la invención es un compuesto de la invención, donde R es Another embodiment of the invention is a compound of the invention, where R is
O OR
- --
- CH 'N ~ CH 'N ~
OÁN I eH, OAN I eH,
H H
Una realización preferible es una composición farmacéutica que comprende al menos un compuesto de la invención, junto con excipientes farmacéuticamente aceptables, para el tratamiento de enfermedades inflamatorias. A preferable embodiment is a pharmaceutical composition comprising at least one compound of the invention, together with pharmaceutically acceptable excipients, for the treatment of inflammatory diseases.
Otra realización más preferible es una composición farmacéutica de la invención en una unidad de dosificación. Y otra realización es que dicha unidad de dosificación sea un comprimido. Another more preferable embodiment is a pharmaceutical composition of the invention in a dosage unit. And another embodiment is that said dosage unit is a tablet.
Una realización preferible es la composición farmacéutica de la invención, que se administra por vía oral. Y otra realización es que dicha composición se administra por vía intravenosa, muscular y/o intramuscular. A preferable embodiment is the pharmaceutical composition of the invention, which is administered orally. And another embodiment is that said composition is administered intravenously, muscularly and / or intramuscularly.
Una realización preferible es una composición farmacéutica que comprende al menos un compuesto de la invención, junto con excipientes farmacéuticamente aceptables y al menos otro agente terapéutico para el tratamiento de enfermedades inflamatorias. A preferable embodiment is a pharmaceutical composition comprising at least one compound of the invention, together with pharmaceutically acceptable excipients and at least one other therapeutic agent for the treatment of inflammatory diseases.
Otra realización más es la composición farmacéutica de la invención, donde dicha enfermedad inflamatoria está causada por bacterias. Y otra realización es que dichas bacterias sean bacterias Gram-negativas. Another embodiment is the pharmaceutical composition of the invention, wherein said inflammatory disease is caused by bacteria. And another embodiment is that said bacteria are Gram-negative bacteria.
Una realización más es la composición farmacéutica de la invención, donde dicha enfermedad inflamatoria es una sepsis. A further embodiment is the pharmaceutical composition of the invention, wherein said inflammatory disease is a sepsis.
Una realización preferible es la composición farmacéutica de la invención, donde dicha enfermedad inflamatoria está seleccionada entre el grupo compuesto por la artritis, infecciones renales, enfermedades pulmonares agudas inducidas por sepsis, síndrome de .distrés respiratorio agudo inducido por sepsis, shock endotóxico o A preferred embodiment is the pharmaceutical composition of the invention, wherein said inflammatory disease is selected from the group consisting of arthritis, renal infections, acute pulmonary diseases induced by sepsis, acute respiratory distress syndrome induced by sepsis, endotoxic shock or
5 .. endotoxemia, peritonitis séptica, hepatitis aguda, disfunción miocárdica aguda inducida por shock séptico y enterecolitis necrotizante. 5 .. endotoxemia, septic peritonitis, acute hepatitis, acute myocardial dysfunction induced by septic shock and necrotizing enterecolitis.
Una realización preferible es el uso de un compuesto derivado de urea pirimidínica de fórmula (1): A preferable embodiment is the use of a pyrimidine urea derivative compound of formula (1):
donde: Res where: Beef
o or
15 donde Ra está seleccionado entre el grupo formado por H, carboxilo, CF3, F, CI, Sr, 1, éter de alquilo C1-C1Q sustituido d no sustituido, éter de alquenilo C2-C10 sustituido o no sustituido, éter de alquinilo CT C1Q sustituido o no sustituido, alquilo C1-C1Q sustituido o no sustituido, alquenilo C2-C1Q sustituido Where Ra is selected from the group consisting of H, carboxyl, CF3, F, CI, Sr, 1, substituted or unsubstituted substituted C1-C1Q alkyl ether, substituted or unsubstituted C2-C10 alkenyl ether, CT alkynyl ether C1Q substituted or unsubstituted, C1-C1Q alkyl substituted or unsubstituted, C2-C1Q alkenyl substituted
o no sustituido, alquinilo C2-C1Q sustituido o no sustituido, cicloalquilo C3-C1Q or unsubstituted, substituted or unsubstituted C2-C1Q alkynyl, C3-C1Q cycloalkyl
20 sustituido o no sustituido, cicloalquenilo CS-C1Q sustituido o no sustituido, cicloalquinilo CS-C1Q sustituido o no sustituido, arilo C6-C10 sustituido o no sustituido, aralquilo Cr C1Q sustituido o no sustituido, heterociclilo de 3-10 miembros de anillo sustituido o no sustituido y heteroarilalquilo sustituido o no sustituido de 2 a 10 átomos de carbono, y ambos heterociclos de 1 a 3 20 substituted or unsubstituted, substituted or unsubstituted CS-C1Q cycloalkenyl, substituted or unsubstituted CS-C1Q cycloalkynyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted Cr C1Q aralkyl, 3-10 substituted ring heterocyclyl or unsubstituted and substituted or unsubstituted heteroarylalkyl of 2 to 10 carbon atoms, and both heterocycles of 1 to 3
25 átomos diferentes al carbono, junto con excipientes farmacéuticamente aceptables para la elaboración de un medicamento. útil para el tratamiento de enfermedades inflamatorias. 25 atoms other than carbon, together with pharmaceutically acceptable excipients for the preparation of a medicament. Useful for the treatment of inflammatory diseases.
Una realización preferible es el uso de la invención, donde R es benceno Otra realización es el uso de la invención, donde R es A preferable embodiment is the use of the invention, where R is benzene. Another embodiment is the use of the invention, where R is
5 Otra realización es el uso de la invención, donde R es eH3 5 Another embodiment is the use of the invention, where R is eH3
I I
~o ~ o
Otra realización es el uso de la invención, donde R es Another embodiment is the use of the invention, where R is
Otra realización es el uso de la invención, donde R es Another embodiment is the use of the invention, where R is
el I~ ó the I ~ ó
el 15 Otra realización es el uso de la invención, donde Res The other embodiment is the use of the invention, where Res
F F
F Otra realización es el uso de la invención, donde R es F Another embodiment is the use of the invention, where R is
Otra realización preferible es el uso de la invención donde dicho medicamento se administra en una unidad de dosificación. Y otra realización es que dicha unidad de 5 dosificación sea un comprimido. Another preferred embodiment is the use of the invention wherein said medicament is administered in a dosage unit. And another embodiment is that said dosage unit is a tablet.
Una realización preferible es el uso de la invención, donde dicho medicamento se administra por vía oral. Y otra realización es que dicho medicamento se administra por vía intravenosa, muscular y/o intramuscular. A preferable embodiment is the use of the invention, wherein said medicament is administered orally. And another embodiment is that said medication is administered intravenously, muscularly and / or intramuscularly.
Una realización preferible es el uso de un compuesto derivado de urea pirimidínica de fórmula (1): A preferable embodiment is the use of a pyrimidine urea derivative compound of formula (1):
donde: 15 Res Where: 15 Res
o or
donde Ra está seleccionado entre el grupo formado por H, carboxilo, CF3, F, CI, Sr, 1, éter de alquilo C1-CIO sustituido o no sustituido, éter de alquenilo 20 Cr CIQ sustituido o no sustituido, éter de alquinilo C2-CIQ sustituido o no sustituido, alquilo C1-CIQ sustituido o no sustituido, alquenilo Cr CIQ sustituido where Ra is selected from the group consisting of H, carboxyl, CF3, F, CI, Sr, 1, substituted or unsubstituted C1-CIO alkyl ether, substituted or unsubstituted C1 Cr alkenyl ether, C2- alkynyl ether Substituted or unsubstituted CIQ, substituted or unsubstituted C1-CIQ alkyl, substituted CI CIQ alkenyl
o no sustituido, alquinilo Cr CIQ sustituido o no sustituido, cicloalquilo C3-CIQ sustituido o no sustituido, cicloalquenilo C5-CIQ sustituido o no sustituido, cicloalquinilo C8-CIQ sustituido o no sustituido, arilo C6-C1O sustituido o no sustituido, aralquilo Cr C10 sustituido o no sustituido, heterociclilo de 3-10 miembros de anillo sustituido o no sustituido y heteroarilalquilo sustituido o no sustituido de 2 a 10 átomos de carbono, y ambos heterociclos de 1 a 3 átomos diferentes al carbono, or unsubstituted, substituted or unsubstituted Cr CIQ alkynyl, substituted or unsubstituted C3-CIQ cycloalkyl, substituted or unsubstituted C5-CIQ cycloalkenyl, substituted or unsubstituted C8-CIQ cycloalkynyl, substituted or unsubstituted C6-C1O aryl, Cr aralkyl C10 substituted or unsubstituted, 3-10 substituted or unsubstituted heterocyclyl ring and substituted or unsubstituted heteroarylalkyl of 2 to 10 carbon atoms, and both heterocycles of 1 to 3 atoms other than carbon,
5 junto con excipientes farmacéuticamente aceptables y al menos otro agente terapéutico para la elaboración de un medicamento útil para el tratamiento de enfermedades inflamatorias. 5 together with pharmaceutically acceptable excipients and at least one other therapeutic agent for the preparation of a medicament useful for the treatment of inflammatory diseases.
Otra realización más es el uso de la invención, donde dicha enfermedad inflamatoria 10 está causada por bacterias. Y otra realización es que dichas bacterias sean bacterias Gram-negativas. Another embodiment is the use of the invention, wherein said inflammatory disease 10 is caused by bacteria. And another embodiment is that said bacteria are Gram-negative bacteria.
Una realización más es el uso de la invención, 'donde dicha enfermedad inflamatoria es una sepsis. A further embodiment is the use of the invention, wherein said inflammatory disease is a sepsis.
Una realización preferible es el uso de la invención, donde dicha enfermedad inflamatoria está seleccionada entre el grupo compuesto por la artritis, infecciones renales, enfermedades pulmonares agudas inducidas por sepsis, síndrome de distrés respiratorio agudo inducido por sepsis, shock endotóxico o endotoxemia, A preferable embodiment is the use of the invention, wherein said inflammatory disease is selected from the group consisting of arthritis, renal infections, acute pulmonary diseases induced by sepsis, acute respiratory distress syndrome induced by sepsis, endotoxic shock or endotoxemia,
20 peritonitis séptica, hepatitis aguda, disfunción miocárdica aguda inducida por shock séptico y enterecolitis necrotizante. 20 septic peritonitis, acute hepatitis, acute myocardial dysfunction induced by septic shock and necrotizing enterecolitis.
La realización más preferible de la invención es una composición administrada por vía intravenosa, que comprende al menos un compuesto derivado de urea 25 pirimidínica de fórmula (1): The most preferable embodiment of the invention is an intravenously administered composition, comprising at least one pyrimidine urea derivative compound of formula (1):
donde: Res CI where: Beef CI
I~ I ~
~~
Jl. Jl.
el he
para el tratamiento de enfermedades inflamatorias. for the treatment of inflammatory diseases.
Otra realización de la invención es un método de tratamiento de una enfermedad inflamatoria que comprende administrar una cantidad efectiva de un compuesto de la invención a un sujeto, preferiblemente humano, en necesidad de dicho tratamiento. Another embodiment of the invention is a method of treating an inflammatory disease that comprises administering an effective amount of a compound of the invention to a subject, preferably human, in need of said treatment.
La figura 1 muestra el efecto de los compuestos 1-7 en la producción de la citoquina pro-inflamatoria IL-8 cuando la línea celular A549 fue estimulada con 300 microgramos/mililitro de LPS de E. Coli y tratada con 100 microMolar de los compuestos 1-7. Se muestran los resultados de las células control (C), células A549 tratadas con LPS de E. Coli (LPS) y células A549 tratadas con LPS de E. Coli más 100 microMolar de los compuestos 1-7 (Compuestos 1-7). En esta figura se representa la simbología estadística: ***p<0.001 vs. C; t p<0.001 vs. LPS. Figure 1 shows the effect of compounds 1-7 on the production of the pro-inflammatory cytokine IL-8 when the A549 cell line was stimulated with 300 micrograms / milliliter of E. coli LPS and treated with 100 microMolar of the compounds 1-7. The results of the control cells (C), A549 cells treated with E. coli LPS (LPS) and A549 cells treated with E. coli LPS plus 100 microMolar of compounds 1-7 (Compounds 1-7) are shown. This figure represents the statistical symbology: *** p <0.001 vs. C; t p <0.001 vs. LPS
La figura 2 muestra el efecto de los compuestos 1-7 en la producción de la citoquina pro-inflamatoria IL-1p cuando la línea celular A549 fue considerada como control (C), tratada con 300 microgramos/mililitro de LPS de E. Coli (LPS) y tratada con 300 microgramos/militro de LPS de E. Coli más 100 microMolar de los compuestos 1-7 (Compuestos 1-7). En esta figura, **p<0.01 vs. C; ,-r p<0.01 vs. LPS. Figure 2 shows the effect of compounds 1-7 on the production of the pro-inflammatory cytokine IL-1p when the A549 cell line was considered as control (C), treated with 300 micrograms / milliliter of LPS from E. Coli ( LPS) and treated with 300 micrograms / milliliter of E. coli LPS plus 100 microMolar of compounds 1-7 (Compounds 1-7). In this figure, ** p <0.01 vs. C; , -r p <0.01 vs. LPS
La figura 3 muestra la bandas correspondientes a los ensayos de Western Blot tras la realización de la extracción de las proteínas totales de células A549 control (C), células A549 bajo el tratamiento con 300 microgramos/mililitro de LPS de E. Coli (LPS) y bajo el tratamiento de 300 microgramos/mililitro de LPS de E. Coli más 100 microMolar de los compuestos 1-7 (Compuestos 1-7). También se muestra la homogeneidad en la concentración proteíca de cada pocillo mediante la Beta-actina. Figure 3 shows the bands corresponding to the Western Blot assays after the extraction of total proteins from control A549 cells (C), A549 cells under treatment with 300 micrograms / milliliter of LPS from E. Coli (LPS) and under the treatment of 300 micrograms / milliliter of LPS of E. Coli plus 100 microMolar of compounds 1-7 (Compounds 1-7). Homogeneity in the protein concentration of each well is also shown by Beta-actin.
La figura 4 muestra los cambios de expresión de las proteínas TLR4 e lK,Ba (normalizados en función del control de carga Beta-actina) cuando células A549 se emplearon como control (C), células A549 bajo el tratamiento con 300 microgramos/mililitro de LPS de E. Coli (LPS) y bajo el tratamiento de 300 microgramos/mililitro de LPS de E. Coli más 100 micro Molar de los compuestos 1-7 (Compuestos 1-7). En esta figura, ** p<0.01 vs. C; ***p<0.001 vs. C; t p<0.001 vs. LPS; ,-r p<0.05 vs. LPS. Figure 4 shows the expression changes of TLR4 and 1K, Ba proteins (normalized according to the Beta-actin load control) when A549 cells were used as control (C), A549 cells under treatment with 300 micrograms / milliliter of E. Coli LPS (LPS) and under the treatment of 300 micrograms / milliliter of E. Coli LPS plus 100 micro Molar of compounds 1-7 (Compounds 1-7). In this figure, ** p <0.01 vs. C; *** p <0.001 vs. C; t p <0.001 vs. LPS; , -r p <0.05 vs. LPS
La figura 5 muestra los valores PS (%) Y la estructura química de diferentes compuestos urea. Figure 5 shows the PS values (%) and the chemical structure of different urea compounds.
5 MODOS DE REALIZACiÓN PREFERENTE Ejemplo 1. Síntesis de los compuestos 1-7 Se sintetizaron los compuestos 1 a 7, derivados de urea pirimidínica, desde los compuestos (pirimidin-4-iloxi) -y (pirimidin-3-il) acetohidrazidas, tal y como se describe en Jakubkiene, V. et al., Synthesis of (pyrimidin4-yloxy)-and (pyrimidin-35 PREFERRED EMBODIMENTS Example 1. Synthesis of compounds 1-7 Compounds 1 to 7, derivatives of pyrimidine urea, were synthesized from the compounds (pyrimidin-4-yloxy) -y (pyrimidin-3-yl) acetohydrazides, such and as described in Jakubkiene, V. et al., Synthesis of (pyrimidin4-yloxy) -and (pyrimidin-3
10 yl)acetyl azides and their rearrangement to carbamates and ureas. Arkivoc 2010 (11) 39-48. Los compuestos puros fueron disueltos en dimetil sulfóxido. 10 yl) acetyl azides and their rearrangement to carbamates and ureas. Arkivoc 2010 (11) 39-48. The pure compounds were dissolved in dimethyl sulfoxide.
Los compuestos 1 a 6 tienen la fórmula: Compounds 1 to 6 have the formula:
siendo R1, R2, Y R3 los grupos que se especifican en la siguiente tabla: R1, R2, and R3 being the groups specified in the following table:
- Compuesto Compound
- R1 R2 R3 R1 R2 R3
- 1 one
- H H H H H H
- 2 2
- eH3 H H eH3 H H
- 3 3
- H H OeH3 H H OeH3
- 4 4
- H H F H H F
- 5 5
- el H el he H he
- 6 6
- H eF3 H H eF3 H
20 El compuesto 7 tiene la fórmula: 20 Compound 7 has the formula:
Ejemplo 2. Actividad anti-microbiana, anti-fúngica y citotóxica de los compuestos Example 2. Anti-microbial, anti-fungal and cytotoxic activity of the compounds
Se determinó la actividad anti-microbiana en bacterias Gram-positivas Staphylococcus aureus (ATCC 29213) y Enterococcus faecalis (ATCC 29212) y bacterias Gram-negativas Escherichia coli (ATCC 35218), Klebsiella pneumoniae (ATCC 700603) y Pseudomonas aeruginosa (ATCC 27853), la actividad anti-fúngica en las especies fúngicas Gandida albicans (ATCC 90028), Gandida glabrata (ATCC 90030), Gandida krusei (ATCC 6258), Gandida nivariensis (5937-63) y Gandida parapsilosis (ATCC 22019) y la actividad citotóxica en las líneas celulares HeLa (línea celular humana de cáncer de cérvix), Ishikawa (línea celular humana de cáncer de endometrio 'uterino), SW1573 (línea celular humana de cáncer epitelial pulmonar), T-47D (línea celular humana de cáncer de pecho), WiDr (Línea celular humana de cáncer de colon) y A549 (línea celular humana epiteliar pulmonar). Anti-microbial activity was determined in Gram-positive bacteria Staphylococcus aureus (ATCC 29213) and Enterococcus faecalis (ATCC 29212) and Gram-negative bacteria Escherichia coli (ATCC 35218), Klebsiella pneumoniae (ATCC 700603) and Pseudomonas aer53inosa (ATCC 700603) , the anti-fungal activity in the fungal species Gandida albicans (ATCC 90028), Gandida glabrata (ATCC 90030), Gandida krusei (ATCC 6258), Gandida nivariensis (5937-63) and Gandida parapsilosis (ATCC 22019) and the cytotoxic activity in HeLa cell lines (human cervical cancer cell line), Ishikawa (human uterine endometrial cancer cell line), SW1573 (human lung epithelial cancer cell line), T-47D (human breast cancer cell line) , WiDr (human colon cancer cell line) and A549 (human pulmonary epithelial cell line).
Las especies bacterianas y fúngicas fueron crecidas en placas de agar a 35-37°C. Después de 24 hora~ de incubación, las especies fueron resuspendidas en solución salina a una concentración aproximadamente de 5 x 105 cfu/ml (unidades formadoras de colonias por mililitro) para las especies bacterianas y 1-5 x 103 para las especies Gandida. La actividad antimicrobiana de los compuestos fue determinada en placas de 96 pocillos usando el medio Mueller Hinton broth para las especies bacterianas y el medio RPMI-1640 tamponado con solución MOPS para las especies fúngicas. Se incluyeron controles estériles y con crecimiento adecuado. Cada compuesto fue testado por duplicado a ocho diferentes soluciones seriadas desde 0,05 a 100 microMolar. Las placas de 96 pocillos fueron incubadas a 35-37°C en una cámara oscura y húmeda. Después de la incubación, las especies fueron precipitadas en 25 microlitros de 50% (peso/volumen) TCA (Ácido tricloroacético) y fijadas durante 60 minutos a 4°C. Después, se llevó a cabo el ensayo colorimétrico descrito en Skehan, Bacterial and fungal species were grown on agar plates at 35-37 ° C. After 24 hour ~ incubation, the species were resuspended in saline solution at a concentration of approximately 5 x 105 cfu / ml (colony forming units per milliliter) for bacterial species and 1-5 x 103 for Gandida species. The antimicrobial activity of the compounds was determined in 96-well plates using Mueller Hinton broth medium for bacterial species and RPMI-1640 medium buffered with MOPS solution for fungal species. Sterile controls with adequate growth were included. Each compound was tested in duplicate to eight different serial solutions from 0.05 to 100 microMolar. The 96-well plates were incubated at 35-37 ° C in a dark and humid chamber. After incubation, the species were precipitated in 25 microliters of 50% (weight / volume) TCA (trichloroacetic acid) and fixed for 60 minutes at 4 ° C. Then, the colorimetric test described in Skehan was carried out,
P. el al., New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Gancer Inst 1990 (82) 1107-1112. Después del ensayo, se produjo la agitación de las placas de 96 pocillos y mediante espectrofotometría (BioTek's PowerWave XS Absorbance Microplate Reader) se obtuvieron valores de densidad óptica. las placas de 96 pocillos correspondientes a las especies bacterianas fueron leídas a 630 nanómetros después de 24 horas de incubación. las placas de 96 pocillos correspondientes a las especies fúngicas fueron leídas a 490 nanómetros después de 48 horas de incubación. Con los resultados obtenidos de la espectrofotometría se obtuvo el valor MCI (mínima concentración inhibitoria) y fue establecido como la concentración del compuesto que inhibió el crecimiento total cuando se comparó con las especies no tratadas. P. el al., New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Gancer Inst 1990 (82) 1107-1112. After the test, the stirring of the 96-well plates was produced and by means of spectrophotometry (BioTek's PowerWave XS Absorbance Microplate Reader) optical density values were obtained. 96-well plates corresponding to bacterial species were read at 630 nanometers after 24 hours of incubation. 96-well plates corresponding to the fungal species were read at 490 nanometers after 48 hours of incubation. With the results obtained from the spectrophotometry, the MCI value (minimum inhibitory concentration) was obtained and was established as the concentration of the compound that inhibited total growth when compared to untreated species.
los compuestos 1 a 7 no presentaban actividad anti-microbiana sobre las bacterias y especies fúngicas ensayadas, ya que todos los compuestos fueron inactivos (valores MCI superiores a 100 microMolar). Compounds 1 to 7 did not exhibit anti-microbial activity on the bacteria and fungal species tested, since all the compounds were inactive (MCI values greater than 100 microMolar).
De la misma manera, las líneas celulares se cultivaron en frascos de cultivo celular de 25 cm2 en medio de cultivo RPMI-1640 suplementado con el 5% de suero fetal bovino y 2 mili Molar de glutamina en un incubador con las siguientes condiciones experimentales: 37°C, 5% de CO2 y 95% de aire húmedo. Células en crecimiento exponencial fueron tripsinizadas y resuspendidas en medio de cultivo que contiene antibióticos (100 unidades de penicilina y 0,1 miligramos de estreptomicina por mililitro). la suspensión celular con un porcentaje superior al 97% de viabilidad por la técnica de exclusión y de tinción Azul Tripán fue contada. Después del contaje, se realizaron las diluciones celulares para su inoculación en placa de 96 pocillos. las células fueron sembradas a una concentración de 1 x 104 (para la línea celular SW1573), 1,5 x 104 (para las líneas celulares Hela, Ishikawa y T-47D) y 2 x 104 (para la línea celular WiDr) células por pocillo. Cada compuesto fue testado por triplicado a diferentes diluciones en el rango de 1-100 microMolar. El tratamiento de los compuestos en los cultivos celulares comenzó un día después de la siembra. El tiempo de incubación de los compuestos fue de 48 horas, después del cual las células fueron precipitadas en 25 microlitros de 50% (pesojvolumen) TCA (Ácido tricloroacético) y fijadas durante 60 minutos a 4°C. Después, se llevó a cabo el ensayo colorimétrico descrito en Skehan, P. et al. la densidad óptica de cada pocillo se leyó a 492 nanómetros empleando el espectrofotómetro BioTek's PowerWave XS Absorbance Microplate Reader. El porcentaje de crecimiento (PG) fue calculado con respecto a las células control no tratadas (C) en cada una de las concentraciones de los compuestos basándose en la diferencia de la densidad óptica al comienzo (To) Y al final de la exposición de los compuestos (T), según las fórmulas descritas en A. Monks et al. Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. J. Natl. Caneer Inst. 1991 (83) 757-766. Se empleó la fórmula PG= 100 X [(T -T o)/(C-To)] cuando T es mayor o igual a To, Y la fórmula PG= 100 X [(T-To)/(To)] cuando T es menor que To. La concentración a la cual PG es +50, O Y -50 representa, respectivamente, un 50% de inhibición del crecimiento (Glso), inhibición total del crecimiento (TGI) y 50% de muerte celular (LCso). Por tanto, un valor PG de O corresponde a la cantidad de células presente en el comienzo de la exposición al compuesto y valores negativos de PG denotan muerte celular neta. In the same way, the cell lines were grown in 25 cm2 cell culture bottles in RPMI-1640 culture medium supplemented with 5% fetal bovine serum and 2 milli Molar glutamine in an incubator with the following experimental conditions: 37 ° C, 5% CO2 and 95% moist air. Exponentially growing cells were trypsinized and resuspended in culture medium containing antibiotics (100 units of penicillin and 0.1 milligrams of streptomycin per milliliter). the cell suspension with a percentage greater than 97% viability by the technique of exclusion and staining Blue Tripán was counted. After counting, cell dilutions were made for inoculation in 96-well plate. cells were seeded at a concentration of 1 x 104 (for the SW1573 cell line), 1.5 x 104 (for the Hela, Ishikawa and T-47D cell lines) and 2 x 104 (for the WiDr cell line) cells per well. Each compound was tested in triplicate at different dilutions in the range of 1-100 microMolar. The treatment of the compounds in cell cultures began one day after sowing. The incubation time of the compounds was 48 hours, after which the cells were precipitated in 25 microliters of 50% (volume weight) TCA (trichloroacetic acid) and fixed for 60 minutes at 4 ° C. Then, the colorimetric assay described in Skehan, P. et al. The optical density of each well was read at 492 nanometers using the BioTek's PowerWave XS Absorbance Microplate Reader spectrophotometer. The percentage of growth (PG) was calculated with respect to the untreated control cells (C) in each of the concentrations of the compounds based on the difference in the optical density at the beginning (To) and at the end of the exposure of the compounds (T), according to the formulas described in A. Monks et al. Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. J. Natl. Caneer Inst. 1991 (83) 757-766. The formula PG = 100 X [(T-T o) / (C-To)] was used when T is greater than or equal to To, and the formula PG = 100 X [(T-To) / (To)] when T is less than To. The concentration at which PG is +50, O and -50 represents, respectively, 50% growth inhibition (Glso), total growth inhibition (TGI) and 50% cell death (LCso). Therefore, a PG value of O corresponds to the amount of cells present at the beginning of exposure to the compound and negative PG values denote net cell death.
Los compuestos empleados en la presente invención no poseían actividad citotóxica sobre las líneas celulares ensayadas ya que todos los compuestos fueron inactivos (valores de inhibición del crecimiento Glso superiores a 100 microMolar). The compounds used in the present invention did not possess cytotoxic activity on the cell lines tested since all the compounds were inactive (Glso growth inhibition values greater than 100 microMolar).
Ejemplo 3. Determinación de los niveles de expresión de IL-8, IL-1p, TLR4 e IKpa. Se cultivó la línea celular A549 (línea celular humana epiteliar pulmonar) en medio de cultivo enriquecido con el 2% de suero fetal bovino durante 24 horas. Posteriormente, se trató el cultivo celular con 300 microgramos/mililitro de LPS de E. Coli (serotipo 055: B5 distribuible comercialmente por la compañía Sigma-Aldrich ®) junto con 100 microMolar de los productos de síntesis descritos en la presente invención durante 18 horas. Pasado el tiempo de tratamiento, se recogió el medio de cultivo en tubos de ensayo Falcon 15 mililitros estériles y se centrifugaron a 1.200 RPM (revoluciones por minuto) durante 10 minutos a 4°C. Después, se recogieron los sobrenadantes de los tubos Falcon 15 mililitros y se añadieron en forma de alícuotas de 500 microlitros en tubos eppendorf 1,5 mililitros estériles. Dichos tubos eppendorf se conservaron en congelación (-80°C) hasta su uso. Example 3. Determination of the expression levels of IL-8, IL-1p, TLR4 and IKpa. The A549 cell line (human pulmonary epithelial cell line) was cultured in culture medium enriched with 2% fetal bovine serum for 24 hours. Subsequently, the cell culture was treated with 300 micrograms / milliliter of E. coli LPS (serotype 055: B5 commercially distributable by the Sigma-Aldrich ® company) together with 100 microMolar of the synthesis products described in the present invention for 18 hours . After the treatment time, the culture medium was collected in sterile Falcon 15 milliliter test tubes and centrifuged at 1,200 RPM (revolutions per minute) for 10 minutes at 4 ° C. Then, the supernatants were collected from the 15 milliliter Falcon tubes and added in the form of 500 microliter aliquots in sterile 1.5 milliliter eppendorf tubes. Said eppendorf tubes were kept frozen (-80 ° C) until use.
Posteriormente, se añadieron 200 microlitos de este medio de cultivo en una placa de ensayo de 96 pocillos y se añadieron los anticuerpos y reactivos del kit comercial de BD Biosciences "Cytometry-based bead array system". El último paso consistió en medir los niveles de expresión de IL-8 e IL-1p mediante ensayo ELlSA a un rango de longitud de 633 nanómetros. Subsequently, 200 microliths of this culture medium were added in a 96-well test plate and the antibodies and reagents of the BD Biosciences commercial kit "Cytometry-based bead array system" were added. The last step was to measure the expression levels of IL-8 and IL-1p by ELlSA assay at a length range of 633 nanometers.
Los niveles de expresión de la proteína de receptor de membrana denominada TLR4 y la proteína inhibidora del factor de transcripción denominado Factor nuclear kappa B (NF-KB) se midieron mediante Western Blot. Para ello, se realizó electroforesis de cada una de las muestras en geles de SDS-PAGE (10%) durante 2 horas. Después de ello, se realizó la transferencia de los geles SDS-PAGE en membranas de difluoruro de polivinilideno (PVDF) y bloqueadas durante 1 hora a temperatura ambiente en el tampón TBST (del inglés Tris-buffered saline Tween-20) más 10% de leche carente de contenido graso. Los anticuerpos anti-TLR4 (Santa Cruz Biotechnology Inc.) y anti-proteína inhibidora NF-KB (Santa Cruz Biotechnology Inc.) se incubaron durante 2 horas en TBST más 5% de leche carente de contenido graso. Después de la incubación durante 2 horas con el anticuerpo primario, se realizó la incubación durante 1 hora con el anticuerpo secundario conjugado con peroxidasa (Goat Anti-rabbit IgG-HRP; Santa Cruz Biotechnology Inc.). La detección de las bandas se llevó a cabo por sistema quimioluminiscente con el kit de GE Healthcare "Amersham ECL Western Blotting Detection Reagent". Posteriormente, se densitometró cada banda proteíca mediante el software público Scion Image. Cada Western Blot se realizó tres veces con las mismas condiciones experimentales nombradas. Expression levels of the membrane receptor protein called TLR4 and the transcription factor inhibitor protein called Kappa B nuclear factor (NF-KB) were measured by Western Blot. For this, electrophoresis of each of the samples was performed in SDS-PAGE gels (10%) for 2 hours. After that, the SDS-PAGE gels were transferred into polyvinylidene difluoride (PVDF) membranes and blocked for 1 hour at room temperature in the TBST buffer (Tris-buffered saline Tween-20) plus 10% of Milk lacking in fat content. The anti-TLR4 (Santa Cruz Biotechnology Inc.) and NF-KB inhibitor anti-protein (Santa Cruz Biotechnology Inc.) antibodies were incubated for 2 hours in TBST plus 5% milk lacking in fat. After incubation for 2 hours with the primary antibody, incubation was performed for 1 hour with the peroxidase-conjugated secondary antibody (Goat Anti-rabbit IgG-HRP; Santa Cruz Biotechnology Inc.). The detection of the bands was carried out by chemiluminescent system with the GE Healthcare kit "Amersham ECL Western Blotting Detection Reagent". Subsequently, each protein band was densitometered using the public software Scion Image. Each Western Blot was performed three times with the same experimental conditions named.
Para comprobar la homogeneidad de la carga proteica en cada uno de los pocillos, las mismas membranas PVDF se re-incubaron con el anticuerpo primario Beta-actina (de la compañía Cell Signaling Technology) durante 2 horas y con el mismo anticuerpo secundario y las mismas condiciones experimentales de los anticuerpos anti-TLR4 y anti-proteína inhibidora del factor de transcripción denominado Factor nuclear kappa B. To check the homogeneity of the protein load in each of the wells, the same PVDF membranes were re-incubated with the primary Beta-actin antibody (from the Cell Signaling Technology company) for 2 hours and with the same secondary antibody and the same experimental conditions of anti-TLR4 and anti-transcription factor inhibitor protein called kappa B nuclear factor.
Todos los datos relacionados con la actividad anti-inflamatoria de los compuestos objeto de la presente invención son expresados como media de los resultados obtenidos para cada compuesto ± error estándar y los análisis estadísticos fueron llevados a cabo empleando los Test de Fisher y la T de Student para datos emparejados y datos no emparejados y empleando el software SPSS (versión 15.0 para Windows). Para los análisis estadísticos de los ensayos Western Blot se empleó la misma metodología pero los datos fueron normalizados en func:ión del control de carga proteíca Beta-actina. Los efectos se consideraron significantes cuando p<0.05. All data related to the anti-inflammatory activity of the compounds object of the present invention are expressed as the average of the results obtained for each compound ± standard error and the statistical analyzes were carried out using the Fisher Test and Student's T for paired data and unpaired data and using the SPSS software (version 15.0 for Windows). For the statistical analyzes of the Western Blot assays, the same methodology was used but the data were normalized according to the beta-actin protein load control ion. The effects were considered significant when p <0.05.
Basados en los ensayos de medición de los niveles de citoquinas pro-inflamatorias en el sobrenadante de la línea celular A549, la figura 1 muestra que los compuestos 1-7 fueron capaces de reducir los niveles de la citoquina pro-inflamatoria IL-8 inducidos bajo el tratamiento de LPS de E. CoN (p<0.001) de una manera significativa (p<0.001). La figura 2 muestra que los compuestos 1-7 objeto de la presente invención fueron capaces de prevenir completamente los efectos producidos por el tratamiento de LPS de E. Coli en los niveles de la citoquina proinflamatoria IL-1,8 (p<0.01). Based on the assays measuring the levels of pro-inflammatory cytokines in the supernatant of the A549 cell line, Figure 1 shows that compounds 1-7 were able to reduce the levels of IL-8 pro-inflammatory cytokine induced under LPS treatment of E. CoN (p <0.001) in a significant way (p <0.001). Figure 2 shows that the compounds 1-7 object of the present invention were able to completely prevent the effects produced by the treatment of E. coli LPS at the levels of the IL-1.8 proinflammatory cytokine (p <0.01).
Basados en los ensayos de Western Blot, las figuras 3 y 4 muestran que la expresión de la proteína TLR4 se incrementó de manera significativa bajo el tratamiento con LPS de E. Coli (p<0.001). Incremento que fue reducido bajo el co-tratamiento de los compuestos 1-7 (p<0.001) y también muestran que la expresión de la proteína inhibidora del factor de transcripción denominado Factor nuclear kappa B se redujo bajo el tratamiento con LPS de E. Coli de manera significativa (p<0.01) y que los compuestos 1-7 objeto de la presente invención produjeron un aumento en los Based on the Western Blot assays, Figures 3 and 4 show that TLR4 protein expression was significantly increased under treatment with E. coli LPS (p <0.001). Increase that was reduced under the co-treatment of compounds 1-7 (p <0.001) and also show that the expression of the transcription factor inhibitor protein called kappa B nuclear factor was reduced under the treatment with E. coli LPS significantly (p <0.01) and that the compounds 1-7 object of the present invention produced an increase in
.niveles de expresión de la proteína lK,8a cuando se comparó este resultado con el tratamiento de LPS de E. Coli (p<0.05). . lK, 8a protein expression levels when this result was compared with the E. coli LPS treatment (p <0.05).
Ejemplo 4. Los compuestos 1 a 7 disminuyen los efectos de LPS producidos en la línea celular A549 Se estimuló la línea celular A549 con 300 microgramos/mililitro de LPS de E. Coli en combinación con cada uno de los compuestos a una dosis simple (100 microMolar). Example 4. Compounds 1 to 7 decrease the effects of LPS produced on the A549 cell line. The A549 cell line was stimulated with 300 micrograms / milliliter of E. Coli LPS in combination with each of the compounds at a single dose (100 microMolar).
El tiempo de incubación de los compuestos fue de18 horas, después del cual las células fueron precipitadas en 25 microlitros de 50% (peso/volumen) TCA (Ácido tricloroacético) y fijadas durante 60 minutos a 4°C. Después, se llevó a cabo el ensayo colorimétrico descrito en Skehan, P. et aL, New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Caneer Inst 1990 (82) 1107-1112. La densidad óptica de cada pocillo se leyó a 492 nanómetros empleando el espectrofotómetro BioTek's PowerWave XS Absorbance Microplate Reader y se determinó el porcentaje de supervivencia (PS) con respecto a las células no tratadas (control negativo) y las células tratadas con 300 microgramos/mililitro de LPS de E. Coli (control positivo, TLPS) de cada concentración de compuesto (T). Por tanto, el cálculo se desarrolló de la siguiente manera: PS= 100 x [(T-TLPS)/(C-TLPS)]. Con este cálculo, un valor de PS de O corresponde a células tratadas solamente con LPS de E. Coli presente al final de la exposición, mientras valores positivos de PS denotan muerte celular neta. The incubation time of the compounds was 18 hours, after which the cells were precipitated in 25 microliters of 50% (weight / volume) TCA (trichloroacetic acid) and fixed for 60 minutes at 4 ° C. Then, the colorimetric assay described in Skehan, P. et al., New colorimetric cytotoxicity assay for anticancer-drug screening was carried out. J Natl Caneer Inst 1990 (82) 1107-1112. The optical density of each well was read at 492 nanometers using the BioTek's PowerWave XS Absorbance Microplate Reader spectrophotometer and the survival percentage (PS) was determined with respect to untreated cells (negative control) and cells treated with 300 micrograms / milliliter of E. coli LPS (positive control, TLPS) of each concentration of compound (T). Therefore, the calculation was developed as follows: PS = 100 x [(T-TLPS) / (C-TLPS)]. With this calculation, a PS value of O corresponds to cells treated only with E. coli LPS present at the end of the exposure, while positive PS values denote net cell death.
En la figura 5 se detallan valores PS correspondientes a diversos derivados de ureas. De los resultados obtenidos, como se muestra en esta figura, se podría concluir que todos los compuestos ensayados poseen la habilidad para disminuir los efectos de LPS producidos en la línea celular A549. Los mejores resultados se 10 alcanzaron con el compuesto 5 donde se alcanzó un valor de PS de 58%. Figure 5 shows PS values corresponding to various urea derivatives. From the results obtained, as shown in this figure, it could be concluded that all the compounds tested possess the ability to decrease the effects of LPS produced in the A549 cell line. The best results were achieved with compound 5 where a PS value of 58% was reached.
Claims (28)
- 20. twenty.
- Uso según reivindicación 19, donde R es benceno. Use according to claim 19, wherein R is benzene.
- 21. twenty-one.
- Uso según reivindicación 19, donde R es Use according to claim 19, wherein R is
- 22. 22
- Uso según reivindicación 19, donde R es Use according to claim 19, wherein R is
- 23. 2. 3.
- Uso según reivindicación 19, donde R es Use according to claim 19, wherein R is
- 26. 26.
- Uso según reivindicación 19, donde Res Use according to claim 19, wherein Res
- 27. 27.
- Uso según la reivindicación 19, donde dicho medicamento se administra en una Use according to claim 19, wherein said medicament is administered in a
- 29. 29.
- Uso según una de las reivindicaciones 27 ó 28, donde dicho medicamento se Use according to one of claims 27 or 28, wherein said medicament is
- 30. 30
- Uso según la reivindicación 19, donde dicho medicamento se administra por vía intravenosa, muscular y/o intramuscular. Use according to claim 19, wherein said medicament is administered intravenously, muscularly and / or intramuscularly.
- 32. 32
- Uso según cualquiera de las reivindicaciones 19 a 31, donde dicha enfermedad inflamatoria está causada por bacterias. Use according to any of claims 19 to 31, wherein said inflammatory disease is caused by bacteria.
- 33. 33.
- Uso según reivindicación 32, donde dichas bacterias son bacterias Gramnegativas. Use according to claim 32, wherein said bacteria are Gram-negative bacteria.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES201031978A ES2385443B2 (en) | 2010-12-28 | 2010-12-28 | USE OF COMPOUNDS DERIVED FROM PYRIMIDINE UREA FOR THE PREPARATION OF A USEFUL MEDICINAL PRODUCT FOR THE TREATMENT OF INFLAMMATORY DISEASES. |
| PCT/ES2011/070822 WO2012089876A1 (en) | 2010-12-28 | 2011-11-25 | Pyrimidine urea‑derived compound for treating inflammatory diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES201031978A ES2385443B2 (en) | 2010-12-28 | 2010-12-28 | USE OF COMPOUNDS DERIVED FROM PYRIMIDINE UREA FOR THE PREPARATION OF A USEFUL MEDICINAL PRODUCT FOR THE TREATMENT OF INFLAMMATORY DISEASES. |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| ES2385443A1 true ES2385443A1 (en) | 2012-07-25 |
| ES2385443B2 ES2385443B2 (en) | 2013-05-16 |
Family
ID=46382347
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES201031978A Expired - Fee Related ES2385443B2 (en) | 2010-12-28 | 2010-12-28 | USE OF COMPOUNDS DERIVED FROM PYRIMIDINE UREA FOR THE PREPARATION OF A USEFUL MEDICINAL PRODUCT FOR THE TREATMENT OF INFLAMMATORY DISEASES. |
Country Status (2)
| Country | Link |
|---|---|
| ES (1) | ES2385443B2 (en) |
| WO (1) | WO2012089876A1 (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1466906A1 (en) * | 1999-03-12 | 2004-10-13 | Boehringer Ingelheim Pharmaceuticals Inc. | Heterocyclic urea and related compounds useful as anti-inflammatory agents |
-
2010
- 2010-12-28 ES ES201031978A patent/ES2385443B2/en not_active Expired - Fee Related
-
2011
- 2011-11-25 WO PCT/ES2011/070822 patent/WO2012089876A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1466906A1 (en) * | 1999-03-12 | 2004-10-13 | Boehringer Ingelheim Pharmaceuticals Inc. | Heterocyclic urea and related compounds useful as anti-inflammatory agents |
Non-Patent Citations (4)
| Title |
|---|
| BRUGEL, T.A. et al. ¿Development of N-2,4-pyrimidine-N-alkyl-N¿-phenylureas as inhibitors of tumor necrosis factor alpha (TNF-¿)synthesis. Part 1.¿ Bioorganic & Medicinal Chemistry Letters 2006,Volumen 16, páginas 3510-3513. [Disponible en línea el 02.05.2006]. Ver página 3510, columna 1, párrafo 1; página 3511, tabla 1. * |
| JAKUBNIENE, V. et al. ¿Synthesis of (pyrimidin-4-yloxy)-and (pyrimidin-3-yl)acetyl azides and their rearrangement to carbamates and ureas¿. ARKIVOC, Volumen 2010, Parte (xi), páginas 39-48. [Publicado el 06.09.2010]. Ver página 40, esquema 1; página 41, esquema 2, página 42, esquema 3. * |
| LATTHE, P.R. et al. ¿Curtius rearrangement reactions of 3-(4-azidocarbonyl) phenylsydnone. Synthesis of 4-(sydnon-3-yl) phenyl carbamates, N-aryl-N¿-[4-(sydnon-3-yl)] phenyl ureas andtheir antimicrobial and insecticidal activities¿. Journal of Chemical Sciences 2006, Volumen 118, Número 3, páginas 249-256. Ver página 249, resumen; página 250, esquema 1. * |
| MAIER, J.A. et al. ¿Development of N-4,6-pyrimidine-N-alkyl-N¿-phenylureas as orally active inhibitors of lymphocytespecific tyrosine kinase.¿ Bioorganic & Medicinal Chemistry Letters 2006, Volumen 16, páginas 3646-3650. [Disponible en línea el 08.05.2006]. Ver página 3646, figura 1; página 3647, tabla 1; página 3650, columna 2, párrafo 3. * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2012089876A1 (en) | 2012-07-05 |
| ES2385443B2 (en) | 2013-05-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chou et al. | Clinical characteristics, antimicrobial susceptibilities, and outcomes of patients with Chryseobacterium indologenes bacteremia in an intensive care unit | |
| CN106687116B (en) | Preparation containing cephalosporins having catechol group | |
| Arif et al. | In vitro and in vivo antimicrobial activities of seeds of Caesalpinia bonduc (Lin.) Roxb. | |
| JP6359013B2 (en) | Inhibitor of apoptosis-related spec-like card protein containing 1,5-D-anhydrofructose | |
| EP2889034B1 (en) | Composition comprising ceftaroline and tobramycin | |
| EP2826473A1 (en) | Novel use of patchoulol | |
| Gan et al. | Nifuroxazide ameliorates pulmonary fibrosis by blocking myofibroblast genesis: a drug repurposing study | |
| Tang et al. | Isochlorogenic acid A alleviates dextran sulfate sodium-induced ulcerative colitis in mice through STAT3/NF-кB pathway | |
| Minjie et al. | Targeting pancreatic cancer cells by a novel hydroxamate-based histone deacetylase (HDAC) inhibitor ST-3595 | |
| ES2385443A1 (en) | Pyrimidine urea-derived compound for treating inflammatory diseases | |
| Joshi et al. | Study of in-vitro antioxidant and antibacterial activity of cow urine from different altitudinal and climatic region of Nepal | |
| CN107236022B (en) | Lipophilic compound conjugate of cell penetrating peptide and application thereof in antibiosis | |
| JP2629166B2 (en) | Antibacterial agent effective against methicillin-resistant Staphylococcus aureus | |
| CN110946870A (en) | Antibacterial pharmaceutical composition and application thereof | |
| CN102887948B (en) | Antibacterial and immunoregulation polypeptide medicine | |
| CN113082026B (en) | Application of artemisinin derivative in preparation of polymyxin antibacterial synergist | |
| Chanda et al. | Global resistance trends and the potential impact of Methicillin Resistant Staphylococcus aureus (MRSA) and its solutions | |
| KR102510514B1 (en) | cell killing agent | |
| KR101396933B1 (en) | The composition of antibacterial complex for animal | |
| Dhanya et al. | Nimbolide from Azadirachta indica and its derivatives plus first-generation cephalosporin antibiotics: a novel drug combination for wound-infecting pathogens | |
| ES2481990B1 (en) | CYCLOPENTENONE COMPOUNDS, PROCEDURE OF OBTAINING AND ITS USE IN THE PREPARATION OF A USEFUL MEDICINAL PRODUCT FOR THE TREATMENT OF INFLAMMATORY DISEASES THAT ARE PROCESSING WITH CELLULAR APOPTOTIC AND FIBROTIC PROCESSES. | |
| Naveed et al. | Assay of different brands of cefadroxil by using spectrophotometric method | |
| ES2408281B1 (en) | USE OF A CHEMICAL COMPOUND DERIVED FROM A 1,2,3,5-TETRAS REPLACED PIRROL IN THE PREPARATION OF A USEFUL MEDICINAL PRODUCT FOR THE TREATMENT OF INFLAMMATORY DISEASES THAT COURSE WITH CELLULAR APOPTOTHICAL PROCESSES. | |
| JP7347915B2 (en) | antibacterial agent | |
| CN118005740B (en) | High-stability high-activity antibacterial polypeptide APH318 and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FG2A | Definitive protection |
Ref document number: 2385443 Country of ref document: ES Kind code of ref document: B2 Effective date: 20130516 |
|
| FD2A | Announcement of lapse in spain |
Effective date: 20180924 |