HK1115132B - N-hydroxyamide derivatives and use thereof - Google Patents
N-hydroxyamide derivatives and use thereof Download PDFInfo
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- HK1115132B HK1115132B HK08110585.2A HK08110585A HK1115132B HK 1115132 B HK1115132 B HK 1115132B HK 08110585 A HK08110585 A HK 08110585A HK 1115132 B HK1115132 B HK 1115132B
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Description
Technical Field
The invention relates to N-hydroxyamide derivatives represented by formula (I), pharmaceutical compositions thereof, a preparation method thereof and applications thereof in treating and/or preventing autoimmune diseases and/or inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, cancers, respiratory diseases and fibrosis. In particular, the present invention relates to N-hydroxyamide derivatives useful for modulating, especially inhibiting, the activity or function of matrix metalloproteinases, especially gelatinases and metalloelastases.
Background
Metalloproteinases belong to the general family of proteases (enzymes) and are named because they rely on a metal ion (zinc) at the active site.
Matrix Metalloproteinases (MMPs) form a subfamily of metalloproteinases, one of their major biological functions is to catalyze the cleavage of connective tissue or extracellular matrix by the ability to hydrolyze various components of the tissue or matrix, such as collagen, gelatin, proteoglycans, fibronectin, and elastin. .
The family of matrix metalloproteinases is further classified according to their function and substrates (Visse et al, 2003, Circ. Res., 92, 827-839), including collagenases (MMP-1, MMP-8, MMP-13 and MMP-18), gelatinases (MMP-2 and MMP-9), stromelysins (MMP-3, MMP-10 and MMP-11), membrane-type MMPs (MT-MMP-1 to MT-MMP-6 and MMP-14, MMP-15, MMP-16, MMP-17, MMP-24 and MMP-25), matrilysins (MMP-7 and MMP-26) and other non-classified MMPs such as metalloelastase (MMP-12), amelysin (MMP-20), epilysin (MMP-28), MMP-19, MMP-22, and MMP-23.
In addition to their connective tissue-degrading role, MMPs are also associated with the biosynthesis of TNF-. alpha.and with posttranslational proteolytic processes, or with the shedding of biologically important membrane proteins (Hooper et al, 1997, Biochem J., 321, 265-279). For example, MMPs act on the local growth and spread of malignant lesions and thus can serve as targets for the development of anti-tumor drugs (Fingleton et al, 2003, Expert opin. the target, 7 (3): 385-. Some diseases such as inflammatory diseases (e.g. Arthritis) (Clark et al, 2003, expert. opin. the target, 7 (1): 19-34 and Liu et al, 2004, Arthritis and Rheumatism (Arthritis and Rheumatism), 50(10), 3112-.
A wide variety of matrix metalloproteinase inhibitors (MMPIs) have been developed (Skiles et al, 2001, Current Medicinal Chemistry (Current Medicinal Chemistry), 8, 425-474; Peterson, 2004, Heart Failure Reviews, 9, 63-79; Henrotin et al, 2002, Expert opin. Ther. patents, 12 (1): 29-43). However, many MMPIs exhibit musculoskeletal syndrome (tendonitis, fibroplasia, mylasia, joint pain), which is a dose-limiting side effect. It has been suggested that inhibition of MMP-1 or MMP-14 may be responsible for these consequences.
Therefore, the development of matrix metalloproteinase inhibitors with defined specificity has become an increasing demand.
Specific inhibitors (particularly for MMP-1) have been reported, including MMP-13 inhibitors (Stotnicki et al, 2003, Current Opinion in drug Discovery and Development, 6 (5): 742-759), MMP-12 inhibitors (WO0)1/83461), MMP-2 and MMP-9 inhibitors (Wada et al, 2002, J.Bio1.chem.45, 219-232).
The high relevance of the metalloprotease pathway in a number of widely spread diseases has exacerbated the need to develop inhibitors including selective inhibitors of MMPs, particularly gelatinases such as MMP-2 and/or MMP-9 and/or MMP-12.
Summary of The Invention
It is an object of the present invention to provide substances suitable for the treatment and/or prevention of diseases involving autoimmune and/or inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, stroke, cancer, preterm labor, endometriosis, respiratory diseases and fibrosis.
It is a further object of the present invention to provide substances suitable for the treatment and/or prevention of multiple sclerosis, rheumatoid arthritis, emphysema, chronic obstructive pulmonary disease and fibrosis.
It is a particular object of the present invention to provide compounds capable of modulating, especially inhibiting, the activity or function of matrix metalloproteinases, especially gelatinases and elastase in mammals, especially humans.
It is a further object of the present invention to provide a novel pharmaceutical formulation for the treatment and/or prevention of a disease selected from the group consisting of autoimmune diseases, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, stroke, cancer, preterm labor, endometriosis, respiratory diseases and fibrosis.
It is a further object of the present invention to provide a process for the preparation of the compounds of the present invention.
It is a final object of the present invention to provide a method for the treatment and/or prevention of a disease selected from the group consisting of autoimmune diseases, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, stroke, cancer, preterm labor, endometriosis, respiratory diseases and fibrosis.
In a first aspect, the present invention provides N-hydroxyamide derivatives of formula (I):
a, R therein1、R2、R3、R4、R5、R6And R7Defined in the detailed description section.
In a second aspect, the present invention provides a compound according to formula (I) for use as a medicament.
In a third aspect, the present invention provides the use of a compound of formula (I) for the preparation of a pharmaceutical composition for the treatment of a disease selected from the group consisting of autoimmune diseases, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, stroke, cancer, preterm labor, endometriosis, respiratory diseases and fibrosis.
In a fourth aspect, the present invention provides a pharmaceutical composition comprising at least one compound of formula (I) and a pharmaceutically acceptable carrier, diluent or excipient.
In a fifth aspect, the present invention provides a method of treatment comprising administering to a patient in need thereof a compound of formula (I).
In a sixth aspect, the present invention provides a method for synthesizing a compound of formula (I).
In a seventh aspect, the present invention provides a compound of formula (IV):
a, R therein1、R2、R3、R4、R5、R6And R7Defined in the detailed description.
Detailed Description
The following paragraphs provide definitions of the various chemical groups that make up the compounds of the present invention and are intended to apply uniformly throughout the specification and claims, unless an otherwise expressly set out definition provides a broader definition.
The term "MMPs" refers to "matrix metalloproteinases". For a recent review of MMPs, see Visse et al, 2003, supra; fingleton et al, 2003, supra; clark et al, 2003, supra and Doherty et al, 2002, Expert Opinion Therapeutic Patents12 (5): 665-707.
Examples of these MMPs include, but are not limited to:
collagenase:are commonly associated with diseases associated with collagen-based tissue cleavage, such as rheumatoid arthritis and osteoarthritis:
MMP-1(also known as collagenase 1, or fibroblast collagenase) and the substrate is collagen I, collagen II, collagen III, gelatin, proteoglycan. Overexpression of this enzyme is thought to be associated with emphysema, hyperkeratosis and atherosclerosis, and overexpression of papillary carcinoma alone.
MMP-8(also known as collagenase 2, or neutrophil collagenase), the substrates are collagen I, collagen II, collagen III, collagen V, collagen VII, collagen IX, gelatin, the overexpression of which leads to non-healing chronic ulcers.
MMP-13(also known as collagenase 3) with collagen I, collagen II, collagen III, collagen IV, collagen IX, collagen X, collagen XIV, fibronectin (fibronectin), gelatin, recently identified as over-expressed in breast cancer alone and associated with rheumatoid arthritis.
Matrix decomposer:
MMP-3(also known as stromelysins)I) The substrates are collagen III, collagen IV, collagen V, collagen IX, collagen X, laminin (laninin), nestin, and overexpression is thought to be associated with atherosclerosis, aneurysm, and restenosis.
Gelatinase-inhibition is considered to have a good effect on cancer, in particular invasion and metastasis.
MMP-2(also known as gelatinase a, 72 kDa gelatinase, basement membrane collagenase, or proteoglycanase) and collagen I, collagen II, collagen IV, collagen V, collagen VII, collagen X, collagen XI, collagen XIV, elastin, fibronectin, gelatin, nestin, are thought to be associated with tumor progression via the specificity of type IV collagen (high expression is observed in solid tumors and is thought to be associated with their growth, invasiveness, neovascular formation and metastasis) and acute pulmonary inflammation and respiratory distress syndrome (Krishna et al, 2004, Expert opin invest drugs, 13 (3): 255-.
MMP-9(also known as gelatinase B, or 92kDa gelatinase) and the substrate is collagen I, collagen III, collagen IV, collagen V, collagen VII, collagen X, collagen XIV, elastin, fibronectin, gelatin, nestin. The above enzymes are thought to be associated with tumor progression via collagen type IV specificity, are released by acid-phagocytic cells in response to exogenous factors such as air pollutants, allergens and viruses, are associated with inflammatory responses in multiple sclerosis (optenakker et al 2003, The Lancet Neurology, 2, 747-. MMP-9 is also thought to be associated with stroke (Horstmann et al, 2003, stroke 34(9), 2165-70).
Unclassified MMPs:
MMP-12(also known as metalloelastase, human macrophage elastase or HME) and the substrate is a fibrous linkProteins, laminins, are thought to play a role in tumor growth inhibition and regulation of inflammation such as multiple sclerosis (Vos et al, 2003, Journal of neuroimaging, 138, 106-.
The expression "MMP-associated disorder" refers to disorders treatable in accordance with the present invention, including all disorders in which the expression and/or activity of at least one MMP needs to be reduced, regardless of the cause of the disorder. Such diseases include, for example, those caused by inappropriate extracellular matrix (ECM) degradation.
Illustrative examples of such MMP-associated disorders include, but are not limited to:
cancers such as breast cancer and solid tumors; inflammatory diseases such as inflammatory bowel disease and neuroinflammation such as multiple sclerosis; pulmonary diseases such as Chronic Obstructive Pulmonary Disease (COPD), emphysema, asthma, acute lung injury, and acute respiratory distress syndrome; dental diseases such as periodontal disease and gingivitis; joint and bone diseases such as osteoarthritis and rheumatoid arthritis; liver diseases such as liver fibrosis, cirrhosis and chronic liver disease; fibrotic diseases such as pulmonary fibrosis, pancreatitis, lupus, glomerulosclerosis, systemic sclerosis skin fibrosis, fibrosis after radiation therapy and cystic fibrosis; vascular diseases such as aortic aneurysm, arteriosclerosis, hypertension, cardiomyopathy and myocardial infarction; restenosis; ophthalmic diseases such as diabetic retinopathy, dry eye syndrome, macular degeneration and corneal ulceration, and degenerative diseases of the central nervous system such as amyotrophic lateral sclerosis.
“C1-C6-alkyl "refers to a monovalent alkyl group having 1 to 6 carbon atoms. The term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-hexyl, and the like. By analogy, "C1-C12-alkyl "refers to a monovalent alkyl group having 1 to 12 carbon atoms, including" C1-C6-an alkyl groupHeptyl, octyl, nonyl, decanoyl, undecanoyl, and dodecanoyl groups, among others. "C1-C10-alkyl "means a monovalent alkyl group having 1 to 10 carbon atoms," C1-C8-alkyl "means a monovalent alkyl group having 1 to 8 carbon atoms," C1-C5-alkyl "refers to a monovalent alkyl group having 1 to 5 carbon atoms.
"Heteroalkyl" refers to C wherein at least one carbon is substituted with a heteroatom selected from O, N or S1-C12Alkyl, preferably C1-C6-alkyl, including 2-methoxyethyl.
"aryl" refers to an unsaturated aromatic carbocyclic group of 6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl). Aryl groups include phenyl, naphthyl, phenanthryl, and the like.
“C1-C6-alkylaryl "means having C1-C6Aryl groups of alkyl substituents, including methylphenyl, ethylphenyl, and the like.
"aryl group C1-C6-alkyl "means C with an aryl substituent1-C6Alkyl groups including 3-phenylpropionyl, benzyl, etc.
"heteroaryl" refers to a monocyclic heteroaromatic or a bicyclic or tricyclic fused ring heteroaromatic group. Specific examples of heteroaryl groups include optionally substituted pyridyl, pyrrolyl, pyrimidinyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, 1, 2, 3-triazolyl, 1, 2, 4-triazolyl, 1, 2, 3-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 5-oxadiazolyl, 1, 3, 4-triazinyl, 1, 2, 3-triazinyl, benzofuranyl, [2, 3-dihydro ] benzofuranyl, isobenzofuranyl, benzothienyl, benzotriazolyl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzimidazolyl, imidazo [1, 2-a ] pyridyl, benzothiazolyl, benzoxazolyl, thiolyl, and the like, Quinolizinyl, quinazolinyl, 2, 3-diazanaphthyl, quinoxalinyl, cinnolinyl, 1, 5-diazanaphthyl, pyrido [3, 4-b ] pyridyl, pyrido [3, 2-b ] pyridyl, pyrido [4, 3-b ] pyridyl, quinolyl, isoquinolyl, tetrazolyl, 5, 6, 7, 8-tetrahydroquinolyl, 5, 6, 7, 8-tetrahydroisoquinolyl, purinyl, pteridinyl, carbazolyl, xanthenyl, or benzoquinolyl.
“C1-C6-alkylheteroaryl "means having C1-C6Heteroaryl groups of alkyl substituents, including methylfuryl and the like.
"heteroaryl group C1-C6-alkyl "means C with a heteroaryl substituent1-C6Alkyl groups including furylmethyl and the like.
“C2-C6-alkenyl "means an alkenyl group preferably having 2 to 6 carbon atoms and having at least 1 or 2 alkenyl unsaturations. Preferred alkenyl groups include vinyl (-CH ═ CH)2) N-2-propenyl (allyl, -CH)2CH＝CH2) And the like.
“C2-C6-alkenylaryl "means having C2-C6Aryl groups of alkenyl substituents, including vinylphenyl and the like.
"aryl group C2-C6-alkenyl "means C with an aryl substituent2-C6Alkenyl groups including phenylvinyl and the like.
“C2-C6-alkenylheteroaryl "means having C2-C6Heteroaryl groups of alkenyl substituents, including vinylpyridinyl and the like.
"heteroaryl group C2-C6-alkenyl "means C with a heteroaryl substituent2-C6Alkenyl groups including pyridylvinyl and the like.
“C2-C6-alkynyl "means an alkynyl group preferably having 2 to 6 carbon atoms and having at least 1 to 2 alkynyl unsaturated bonds. Preferred alkynyl groups includeAlkynyl (-CH. ident.CH), propynyl (-CH)2C.ident.CH) and the like.
“C3-C8-cycloalkyl "refers to a saturated carbocyclic group of 3 to 8 carbon atoms having a single ring (e.g., cyclohexyl) or multiple condensed rings (e.g., norbornyl). C3-C8Cycloalkyl includes cyclopentyl, cyclohexyl, norbornyl, and the like.
"Heterocycloalkyl" means C as defined above3-C8Cycloalkyl in which up to 3 carbon atoms are substituted by a heteroatom selected from O, S, NR, R being defined as hydrogen or methyl. Heterocycloalkyl groups include pyrrolidine, piperidine, piperazine, morpholine, tetrahydrofuran, and the like.
“C1-C6-alkylcycloalkyl "denotes a compound having C1-C6C of an alkyl substituent3-C8Cycloalkyl groups, including methylcyclopentyl and the like.
"cycloalkyl group C1-C6-alkyl "means having C3-C8-C of a cycloalkyl substituent1-C6Alkyl groups including 3-cyclopentylpropyl and the like.
“C1-C6-Alkylheterocycloalkyl "means having C1-C6Heterocycloalkyl groups of alkyl substituents, including 1-methylpiperazine and the like.
"Heterocycloalkyl radical C1-C6-alkyl "means C with a heterocycloalkyl substituent1-C6Alkyl groups including 4-methylpiperidinyl and the like.
"carboxy" refers to the group-C (O) OH.
"carboxy group C1-C6-alkyl "means C with a carboxyl substituent1-C6Alkyl groups including 2-carboxyethyl and the like.
"acyl" refers to the group-C (O) R, wherein R includes "C1-C12-alkyl ", preferably" C1-C6-alkyl "," aryl "," heteroaryl "-, or,“C3-C8-cycloalkyl "," heterocycloalkyl "," aryl C1-C6-alkyl "," heteroaryl C1-C6-alkyl group "," C3-C8-cycloalkyl group C1-C6-alkyl "or" heterocycloalkyl C1-C6-an alkyl group ".
"acyl group C1-C6-alkyl "means C with an acyl substituent1-C6Alkyl groups including acetyl, 2-acetylethyl and the like.
"acylaryl" refers to an aryl group having an acyl substituent, including 2-acetylphenyl and the like.
"acyloxy" refers to the group-OC (O) R, where R includes H, "C1-C6-alkyl group "," C2-C6Alkenyl group "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"acyloxy group C1-C6-alkyl "means C with an acyloxy substituent1-C6Alkyl groups including ethyl propionate and the like.
"alkoxy" refers to the group-O-R, where R includes "C1-C6-alkyl "or" aryl "or" heteroaryl "or" aryl C1-C6-alkyl "or" heteroaryl C1-C6-an alkyl group ". Preferred alkoxy groups include, for example, methoxy, ethoxy, phenoxy and the like.
"alkoxy group C1-C6-alkyl "means having C1-C6Alkoxy groups of alkyl substituents, including methoxy, methoxyethyl, and the like.
"alkoxycarbonyl" refers to the group-C (O) OR, where R includes H, "C1-C6-alkyl "or" aryl "or" heteroaryl "or" aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl "or" heteroalkyl ".
"alkoxycarbonyl group C1-C6-alkyl "means C having an alkoxycarbonyl substituent1-C5Alkyl groups including 2- (benzyloxycarbonyl) ethyl and the like.
"aminocarbonyl" refers to the group-C (O) NRR 'where R, R' independently includes hydrogen or C1-C6-alkyl or aryl or heteroaryl or "aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl "including N-phenylcarboxamide.
"aminocarbonyl group C1-C6-alkyl "means C with an aminocarbonyl substituent1-C6Alkyl groups including 2- (dimethylaminocarbonyl) ethyl, N-ethylacetamide, N-diethylacetamide, and the like.
"acylamino" refers to the group-NRC (O) R ', wherein each R, R' is independently hydrogen, "C1-C6-alkyl group "," C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6Alkyl groups ".
"acylamino C1-C6-alkyl "means C with an acylamino substituent1-C6Alkyl groups including 2- (propionylamino) ethyl and the like.
"ureido" means a radical-NRC (O) NR 'R "where R, R', R" are independently hydrogen, "C1-C6-alkyl group "," C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-alkyl ", and wherein R' and R" together with the nitrogen atom to which they are attached may optionally form a 3-8 membered heterocycloalkyl ring.
"ureido group C1-C6-alkyl "means C having a ureido substituent1-C6Alkyl groups including 2- (N' -methylureido) ethyl and the like.
"Carbamate" refers to the group-NRC (O) OR ', wherein R, R' is independently hydrogen, "C1-C6-alkyl group "," C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," C1-C6-alkylaryl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"amino" refers to the group-NRR 'where R, R' is independently hydrogen or "C1-C6-alkyl "or" aryl "or" heteroaryl "or" C1-C6-alkylaryl "or" C1-C6-alkylheteroaryl "or" cycloalkyl "or" heterocycloalkyl ", and wherein R' and R" together with the nitrogen atom to which they are attached may optionally form a 3-8 membered heterocycloalkyl ring.
"amino group C1-C6-alkyl "means C with an amino substituent1-C5Alkyl groups including 2- (1-pyrrolidinyl) ethyl and the like.
"ammonium" refers to a positively charged group-N+RR 'R' where R, R ', R' are independently "C1-C6-alkyl "or" C1-C6-alkylaryl "or" C1-C6-alkylheteroaryl "or" cycloalkyl "or" heterocycloalkyl ", and wherein R and R', together with the nitrogen atom to which they are attached, may optionally form a 3-8 membered heterocycloalkyl ring.
"ammonium C1-C6-alkyl "means C with an ammonium substituent1-C6Alkyl groups including 1-ethylpyrrolidinium and the like.
"halogen" refers to fluorine, chlorine, bromine and iodine atoms.
"Sulfonyloxy" refers to the group-OSO2-R, wherein R is selected from H, "C1-C6-alkyl ", halogen-substituted" C1-C6Alkyl radicals "e.g. -OSO2-CF3、“C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," arylC2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"Sulfonyloxy group C1-C6-alkyl "means C having a sulfonyloxy substituent1-C6Alkyl groups including 2- (methylsulfonyloxy) ethyl and the like.
"Sulfonyl" refers to the group "-SO2-R ", wherein R is selected from H," aryl "," heteroaryl "," C1-C6-alkyl ", halogen-substituted" C1-C6Alkyl radicals "such as-SO2-CF3、“C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"Sulfonyl group C1-C6-alkyl "means C having a sulfonyl substituent1-C5Alkyl groups including 2- (methylsulfonyl) ethyl and the like.
"sulfinyl" refers to the group "-S (O) R", wherein R is selected from H, "C1-C6-alkyl ", halogen-substituted" C1-C6Alkyl radicals "such as-SO-CF3、“C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl "And aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"sulfinyl group C1-C6-alkyl "means C having a sulfinyl substituent1-C6Alkyl groups including 2- (methylsulfinyl) ethyl and the like.
"Thioalkyl" refers to the group-S-R, where R includes H, "C1-C6-alkyl ", halogen-substituted" C1-C6Alkyl radicals "such as-SO-CF3、“C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
Preferred sulfanyl groups include methylsulfanyl, ethylsulfanyl, and the like.
"Thioalkyl radical C1-C6-alkyl "means C with a sulfanyl substituent1-C5Alkyl groups including 2- (ethylsulfanyl) ethyl and the like.
"Sulfonylamino" refers to the group-NRSO2-R ', wherein each R, R' independently comprises H, "C1-C6-alkyl group "," C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl ",aryl, heteroaryl, aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"Sulfonylamino C1-C6-alkyl "means C having a sulfonylamino substituent1-C6Alkyl groups including 2- (ethylsulfonylamino) ethyl and the like.
"aminosulfonyl" refers to the group-SO2-NRR ', wherein each R, R' independently comprises H, "C1-C6-alkyl group "," C2-C6-alkenyl "," C2-C6-alkynyl "," C3-C8-cycloalkyl "," heterocycloalkyl "," aryl "," heteroaryl "," aryl C1-C6-alkyl "or" heteroaryl C1-C6-alkyl group "," aryl group C2-C6-alkenyl "," heteroaryl C2-C6-alkenyl "," aryl C2-C6-alkynyl "," heteroaryl C2-C6-alkynyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-an alkyl group ".
"Aminosulfonyl group C1-C6-alkyl "means C having an aminosulfonyl substituent1-C6Alkyl groups including 2- (cyclohexylaminosulfonyl) ethyl and the like.
"substituted or unsubstituted": unless otherwise limited by the definition of each substituent, groups described above, such as "alkenyl", "alkynyl", "aryl", "heteroaryl", "cycloalkyl", "heterocycloalkyl", and the like, may be optionally substituted with 1 to 5 substituents selected from: "C1-C6-alkyl group "," C2-C6-alkenyl "," C2-C6-alkynyl "," cycloalkyl "," heterocycloalkyl "," aryl C1-C6-alkyl "," heteroaryl C1-C6-alkyl "," cycloalkyl C1-C6-alkyl "," heterocycloalkyl C1-C6-alkyl "," amino "," ammonium "," acyl "," acyloxy "," acylamino "," aminocarbonyl "," alkoxycarbonyl "," ureido "," aryl "," carbamate "," heteroaryl "," sulfinyl "," sulfonyl "," alkoxy "," sulfanyl "," halogen "," carboxy ", trihalomethyl, cyano, hydroxy, mercapto, nitro, and the like.
"pharmaceutically acceptable salt or complex" refers to a salt or complex of a compound of formula (I) as defined below. Examples of such salts include, but are not limited to, base addition salts formed by reacting a compound of formula (I) with an organic or inorganic base such as a hydroxide, carbonate or bicarbonate of a metal cation selected, for example, from the alkali metals (sodium, potassium or lithium) or alkaline earth metals (e.g., calcium or magnesium); or a base addition salt formed by reaction with an organic primary, secondary or tertiary alkylamine. Ammonium salts of amine salts derived from methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, morpholine, N-Me-D-glucamine, N' -bis (phenylmethyl) -1, 2-ethylenediamine, tromethamine, ethanolamine, diethanolamine, 1, 2-ethylenediamine, N-methylmorpholine, procaine, piperidine, piperazine and the like are considered to be within the scope of the present invention.
Acid addition salts with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like); and acid addition salts with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoic acid, tannic acid, 4' -methylenebis (3-hydroxy-2-naphthoic acid), alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid, are also included.
"pharmaceutically active derivative" refers to any compound that, when administered to a recipient, is capable of providing, directly or indirectly, the activity disclosed herein. The term "indirectly" also includes prodrugs that are convertible to a pharmaceutically active form by endogenous enzymes or metabolism. The prodrug consists of the active drug compound itself and a chemical masking group. Such masking groups may be cyclopropanone compounds of formula (I ') wherein Y is a methyl group or hydrogen, Y' is a methyl group, C2-C4Alkyl, phenyl, benzyl, optionally substituted with 1 to 3 substituents selected from the group consisting of: c1-C4Alkyl radical, C1-C4Alkoxy, hydroxy, amino, methylamino, dimethylamino, chloro, and fluoro; A. r1、R2、R4、R5、R6And R7Defined in the detailed description.
"enantiomeric excess" (ee) refers to the production of a product wherein one enantiomeric excess (ee) is at least about 52% by asymmetric synthesis, such as synthesis involving non-racemic starting materials and/or reagents, or synthesis comprising at least one enantioselective step.
"Interferon" or "IFN" as used herein, includes any molecule defined in the literature, including, for example, any of the types of IFNs described in the "background" section above. Specifically, IFN- α, IFN- β and IFN- γ are included within the above definitions. A preferred IFN of the invention is IFN- β. IFN- β applicable in accordance with the invention is commercially available, e.g., Rebif(Serono)、Avonex(Biogen) or Betaferon(Schering)。
The term "interferon-beta (IFN-beta or IFN-beta)" as used herein includes human fibroblast interferon, in particular of human origin, obtained by isolation from a biological fluid or by recombinant DNA techniques from prokaryotic or eukaryotic host cells, as well as salts, functional derivatives, mutants, analogs and active fractions thereof. Preferably, IFN- β refers to recombinant interferon β -1 a.
IFN- β s applicable in accordance with the invention are commercially available, e.g., Rebif(Serono)、Avonex(Biogen) or Betaferon(Schering). Human interferons are also preferably used in accordance with the present invention. The term interferon as used herein is intended to include salts, functional derivatives, mutants, analogs and active moieties thereof.
A recent development in interferon therapy is the use of Rebif(recombinant interferon-beta) treatment of Multiple Sclerosis (MS), represents a significant advance in therapeutic approaches. RebifIs Interferon (IFN) - β 1a, produced by a mammalian cell line. It has been determined that interferon beta-1 a administered subcutaneously three times a week has therapeutic effects on relapsing-remitting multiple sclerosis (RRMS). Interferon beta-1 a has beneficial effects on long-term MS disease course, it reduces the number and severity of relapses, and reduces disease burden and disease activity as determined by MRI.
The dosage of IFN- β for the treatment of relapsing-remitting MS according to the invention depends on the type of IFN- β used.
According to the invention, when the IFN is a recombinant IFN- β 1b (commercially available under the trademark Betaseron) produced by E.coli) In this case, it is preferred to administer the drug subcutaneously every other day at a dose of about 250 μ g or 8 MIU to 9.6 MIU per person.
According to the present invention, when IFN is recombinant IFN-. beta.1a (commercially available under the trademark Avonex) produced by Chinese hamster ovary cells (CHO cells)) In this case, it is preferably administered intramuscularly once a week at a dose of about 30-33. mu.g or 6 MIU-6.6MIU per person.
According to the invention, when IFN is produced by Chinese hamster ovary cells (CHO cells) recombinant IFN-beta 1a (commercially available under the trademark Rebif)) In this case, it is preferred to administer subcutaneously three times a week (TIW) at a dose of about 22-44 μ g or 6 MIU-12MIU per person.
The compounds of the present invention also include pharmaceutically acceptable salts thereof. Preferred pharmaceutically acceptable salts of formula (I) are acid addition salts with pharmaceutically acceptable acids such as hydrochloride, hydrobromide, sulphate or bisulphate, phosphate or hydrogenphosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate, methanesulphonate, benzenesulphonate and p-toluenesulphonate.
It has now been found that the compounds of the present invention are modulators of matrix metalloproteinases, particularly gelatinases and elastase including MMP-2 and/or MMP-9 and/or MMP-12. When matrix metalloproteases are inhibited by the compounds of the invention, the MMP(s) inhibited are unable to exert their enzymatic, biological and/or pharmacological effects. The compounds of the invention are therefore useful in the treatment and prevention of autoimmune and/or inflammatory diseases, cardiovascular diseases, preterm labor, endometriosis, neurodegenerative diseases, stroke, cancer, respiratory diseases and fibrosis.
In one embodiment, the present invention provides derivatives of formula (I) and optically active forms thereof such as enantiomeric, diastereomeric and racemic forms, and pharmaceutically acceptable salts thereof.
Wherein:
a is selected from-C, (B) -and N;
b is H or B and R5Or R7One of which is bonded;
R1is selected from H; optionally substituted C1-C6An alkyl group;
optionally substituted C2-C6An alkenyl group; optionally substituted C2-C6An alkynyl group;
optionally substituted C3-C8-cycloalkyl, including cyclohexyl;
optionally substituted heterocycloalkyl;
optionally substituted aryl groups including optionally substituted phenyl such as phenyl, fluorophenyl (e.g. 2-fluorophenyl, 4-fluorophenyl, 3-chlorophenyl), chlorophenyl (e.g. 2-chlorophenyl, 4-chlorophenyl), methoxyphenyl (e.g. 4-methoxyphenyl), ethoxyphenyl (e.g. 4-ethoxyphenyl), cyanophenyl (e.g. 2-cyanophenyl), trifluoromethylphenyl (e.g. 4-trifluoromethoxyphenyl), biphenyl (e.g. 4-biphenyl) and 4-chloro-2-fluorophenyl, 2-fluoro-5-methoxyphenyl;
optionally substituted heteroaryl, including optionally substituted pyridyl, such as pyridyl, methylpyridyl (such as 4-methylpyridin-2-yl, 6-methylpyridin-2-yl), chloropyridyl (such as 6-chloropyridin-2-yl, 5-chloropyridin-2-yl, 3, 5-dichloropyridin-4-yl), trifluoromethylpyridyl (such as 3- (trifluoromethyl) pyridin-2-yl, 4- (trifluoromethyl) pyridin-2-yl, 5- (trifluoromethyl) pyridin-2-yl), cyanopyridyl (such as 5-cyanopyridin-2-yl), phenylpyridyl (such as 5-phenylpyridin-2-yl) and optionally substituted fused pyridyl (such as 4- [ 6-methyl-2- (trifluoromethyl) quinoline- 4-yl)); including optionally substituted pyrazinyl (e.g. 4-pyrazin-2-yl); including optionally substituted thiadiazolyl groups such as 3-phenylthiadiazolyl (e.g., 3-phenyl-1, 2, 4-thiadiazol-5-yl); including optionally substituted pyrimidinyl (e.g., 4-pyrimidinyl-2-yl); including optionally substituted oxadiazolyl groups such as 5-phenyl-1, 2, 4-oxadiazol-3-yl, 4-pyridin-4-yl-1, 2, 4-oxadiazol-3-yl and 5- (4-fluorophenyl) -1, 3, 4-oxadiazol-2-yl;
optionally substituted C3-C8-cycloalkyl group C1-C6An alkyl group;
optionally substituted heterocycloalkyl C1-C6Alkyl groups including 2-morpholin-4-ylethyl;
optionally substituted heteroaryl C1-C6Alkyl, including 2-thienylethyl;
optionally substituted amino, including optionally substituted phenylamino (e.g., phenylamino, 3-methoxyphenylamino, 3- (dimethylamino) phenylamino, 4-ethoxyphenylamino), heteroarylamino (e.g., 4-trifluoromethyl) pyrimidin-2-yl, 3-aminopyridin-2-yl); and
optionally substituted alkoxy, including 4- (pyridin-2-yloxy), 4- (trifluoromethyl) phenoxy, 2-chlorophenoxy;
R2is H;
R3selected from H, optionally substituted C1-C6Alkyl, optionally substituted C2-C6Alkenyl and optionally substituted C2-C6An alkynyl group;
R4、R5、R6and R7Independently selected from H; optionally substituted C1-C6Alkyl, including methyl; optionally substituted C2-C6An alkenyl group; optionally substituted C2-C6An alkynyl group; or R4And R7May together form a CH2Linkage, e.g. to form a 2, 5-azabicyclo [2.2.1 ] ring with a piperazine ring]The hept-2-yl ring.
In a preferred embodiment, the present invention provides a derivative of formula (I), wherein R1Selected from optionally substituted aryl and optionally substituted heteroaryl.
In a more preferred embodiment, the present invention provides a derivative of formula (I), wherein R1Is optionally substituted aryl such as optionally substituted phenyl, including fluorophenyl (e.g., 4-fluorophenyl), methoxyphenyl (e.g., 4-trifluoromethoxyphenyl), and biphenyl (e.g., 4-biphenyl-4-yl).
In another preferred embodiment, the present invention provides a derivative of formula (I), wherein R is3Is H.
In another preferred embodiment, the present invention provides a derivative of formula (I), wherein R is5、R6And R7Is H.
In another preferred embodiment, the present invention provides a derivative of formula (I), wherein R is4Selected from H and optionally substituted C1-C6Alkyl groups, including methyl.
In a further embodiment, the present invention provides a derivative of formula (I), wherein R is4Is H.
In a further embodiment, the present invention provides a derivative of formula (I), wherein R is4Is methyl.
In another preferred embodiment, the present invention provides a derivative of formula (I) wherein a is N.
In another preferred embodiment, the present invention provides a derivative of formula (I), wherein R is1Is optionally substituted aryl, including optionally substituted phenyl; r3、R5、R6And R7Is H; r4Selected from H and methyl; a is N.
The compounds of the invention include in particular compounds selected from the following group:
(2R) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
(2S) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
(2S) -N, 2-dihydroxy-4- { (2R) -2-methyl-4- [4- (trifluoromethoxy) phenyl ] piperazin-1-yl } -4-oxobutanamide;
(2S) -4- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide.
In another embodiment of the present invention, there are provided N-hydroxyamide derivatives according to formula (I) for use as a medicament.
In another embodiment of the present invention, a pharmaceutical composition is provided comprising at least one N-hydroxyamide derivative of the present invention and a pharmaceutically acceptable carrier, diluent or excipient.
In another embodiment of the present invention, there is provided a use of an N-hydroxyamide derivative represented by formula (I) for the preparation of a medicament for the prophylaxis and/or treatment of a disease selected from autoimmune diseases, inflammatory diseases, stroke, cardiovascular diseases, neurodegenerative diseases, cancer, premature labor, endometriosis, respiratory diseases and fibrosis, including multiple sclerosis, inflammatory bowel diseases, rheumatoid arthritis, emphysema, Chronic Obstructive Pulmonary Disease (COPD), liver cirrhosis and fibrosis, including liver and lung, pancreatic fibrosis and liver fibrosis.
In another embodiment of the invention, there is provided the use of an N-hydroxyamide derivative of formula (I) for the preparation of a pharmaceutical preparation for modulating, in particular inhibiting, the activity of a matrix metalloproteinase. In particular, there is provided a use according to the invention wherein the matrix metalloproteinase is selected from MMP-2, MMP-9 and MMP-12. Preferably, the compounds according to the invention are selective inhibitors of metalloproteases selected from MMP-2, MMP-9 and/or MMP-12 over MMP-1.
In another embodiment, the invention provides a method of treating and/or preventing a disease in a patient in need thereof, comprising the step of administering a compound of formula (I), wherein the disease is selected from the group consisting of autoimmune diseases, inflammatory diseases, cardiovascular diseases, premature labor, endometriosis, neurodegenerative diseases, stroke, cancer, respiratory diseases and fibrosis, including multiple sclerosis, rheumatoid arthritis, emphysema, Chronic Obstructive Pulmonary Disease (COPD) and fibrosis, including liver cirrhosis and fibrosis, including lung, pancreas and liver fibrosis.
In another embodiment, the invention provides a process for the preparation of the N-hydroxyamide derivatives of the invention, comprising reacting a compound of formula (IV) with H2NO-R8The step of derivative reaction:
a, R therein1、R2、R4、R5、R6And R7As defined above, R8Selected from H and a protecting group such as tert-butyl, benzyl, trialkylsilyl, tetrahydropyranyl.
In yet another embodiment, the invention provides a process for the preparation of the N-hydroxyamide derivatives of the invention, optionally further comprising a deprotection step (when R is8When not H, removing R8)。
In another embodiment, the invention provides a compound of formula (IV):
a, R therein1、R2、R4、R5、R6And R7As defined above.
In yet another embodiment, the present invention provides a compound of formula (IV) selected from the group consisting of:
(5R) -5- {2- [4- (4-fluorophenyl) piperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one;
(5S) -5- {2- [4- (4-fluorophenyl) piperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one;
(5S) -2, 2-dimethyl-5- [2- ((2R) -2-methyl-4- {4- [ (trifluoromethyl) oxy ] phenyl } piperazin-1-yl) -2-oxoethyl ] -1, 3-dioxolan-4-one;
(5S) -5- {2- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one.
The compounds of the invention were named according to the standard adopted by the program "ACD/Name" of Advanced Chemistry Development Inc., ACD/Labs (7.06 Release).
The compounds of formula (I) are useful in the treatment and/or prevention of autoimmune diseases, inflammatory diseases, cardiovascular diseases, premature birth, endometriosis, neurodegenerative diseases, stroke, cancer, premature birth, endometriosis, respiratory diseases and fibrosis, including multiple sclerosis, rheumatoid arthritis, emphysema, chronic obstructive pulmonary disease and fibrosis, including liver cirrhosis and fibrosis, including liver and lung, pancreatic fibrosis and liver fibrosis.
In another embodiment, the compounds of the invention can be used for the treatment of autoimmune diseases, in particular demyelinating diseases such as multiple sclerosis, alone or in combination with a combination useful for the treatment of autoimmune diseases, wherein the combination is selected from, for example, the following compounds:
(a) interferons, e.g. pegylated or non-pegylated interferons, e.g. administered by subcutaneous, intramuscular or oral routes, preferably interferon beta;
(b) glatiramer (Glatiramer), e.g., in the form of its acetate salt;
(c) immunosuppressive agents with or without antiproliferative/antitumor activity, such as mitoxantrone, methotrexate, azathioprine, cyclophosphamide, or steroids such as methylprednisolone, prednisone or dexamethasone, or steroid-secreting drugs, such as ACTH;
(d) adenosine deaminase inhibitors, such as cladribine;
(e) an inhibitor of VCAM-1 expression or an antagonist of a ligand thereof, such as an antagonist of the α 4/β 1 integrin VLA-4 and/or α -4- β -7 integrin, e.g., natalizumab (ANTEGRENO).
Further combinations of drugs such as anti-inflammatory drugs (especially demyelinating diseases such as multiple sclerosis) are described below:
one particular anti-inflammatory drug is Teriflunomide (Teriflunomide) described in WO 02/080897.
Another specific anti-inflammatory drug is Fingolimod described in EP-727406, WO2004/028251 and WO 2004/028251.
Another specific anti-inflammatory drug is Laquinimod described in WO 99/55678.
Another specific anti-inflammatory drug is Tensirolimus described in WO 02/28866.
Another specific anti-inflammatory drug is Xaliprodene described in WO 98/48802.
Another specific anti-inflammatory drug is the descara pirfenidone described in WO 03/068230.
Another specific anti-inflammatory drug is benzothia as described in WO 01/47920
An azole derivative.
Another specific anti-inflammatory drug is one of the hydroxamic acid derivatives described in WO 03/070711.
Another specific anti-inflammatory drug is MLN3897 described in WO 2004/043965.
Another specific anti-inflammatory drug is CDP323 described in WO 99/67230.
Another specific anti-inflammatory drug is simvastatin, described in WO 01/45698.
Another specific anti-inflammatory drug is aminopyridine (Fampridine) described in US 5,540,938.
The compounds of the present invention also include tautomers thereof, geometrical isomers thereof, optically active forms thereof such as enantiomers, diastereomers and racemic forms thereof, and pharmaceutically acceptable salts thereof. Preferred pharmaceutically acceptable salts of formula (VI) are acid addition salts with pharmaceutically acceptable acids such as hydrochloride, hydrobromide, sulphate or bisulphate, phosphate or hydrogenphosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate, methanesulphonate, benzenesulphonate and p-toluenesulphonate.
The derivatives exemplified in the present invention can be prepared from readily available starting materials using the following general methods and procedures. It should be recognized that while typical or preferred experimental conditions (i.e., reaction temperatures, times, moles of reagents, solvents, etc.) are given, other experimental conditions may be used unless otherwise indicated. Optimum reaction conditions will vary with the particular reagents or solvents used, but such conditions can be determined by one skilled in the art using routine optimization procedures.
When used as a medicament, the compounds of the invention are typically administered in the form of a pharmaceutical composition. Accordingly, pharmaceutical compositions comprising a compound of the invention and a pharmaceutically acceptable carrier, diluent or excipient are also within the scope of the invention. Those skilled in the art are aware of all such carriers, diluents or excipients which are suitable for formulating pharmaceutical compositions.
The compounds of the present invention may be incorporated in the dosage form of pharmaceutical compositions and unit doses thereof, in solid form (e.g., tablets or filled capsules) or in liquid form (e.g., solutions, suspensions, emulsions, elixirs, or capsules filled with such liquids), with the usual adjuvants, carriers, diluents, or excipients for oral use; or a sterile injectable solution for parenteral administration (including subcutaneous administration). Such pharmaceutical compositions and unit dosage forms may contain the ingredients in conventional proportions, with or without other active compounds or active ingredients, and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed.
Pharmaceutical compositions containing the compounds of the invention may be prepared by methods well known in the pharmaceutical arts and include at least one active compound. Typically, the compounds of the present invention are administered in a pharmaceutically effective amount. The amount of the compound actually administered will generally be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of each patient, the severity of the patient's symptoms, and the like.
The pharmaceutical compositions of the present invention may be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. Compositions for oral administration may take the form of large liquid solutions or suspensions, or large powders. More typically, however, the compositions will be presented in unit dosage form to facilitate accurate administration. The term "unit dosage form" refers to physically discrete units suitable for use as a single dose in humans or other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, pre-metered ampoules or syringes of liquid compositions, or pills, tablets, capsules and the like of solid compositions. In such compositions, the derivatives of the invention are generally a minor component (about 0.1 to 50% by weight or preferably about 1 to 40% by weight), the remainder being various excipients or carriers and processing aids which assist in forming the desired dosage form.
Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous vehicles containing buffers, suspending and dispersing agents, colors, flavors, and the like.
The solid form may include, for example, any of the following ingredients or compounds of similar nature: a binder (such as microcrystalline cellulose, tragacanth or gelatin), an excipient (such as starch or lactose), a disintegrant (such as alginic acid, Primogel, or corn starch), a lubricant (such as magnesium stearate), a glidant (such as colloidal silicon dioxide), a sweetening agent (such as sucrose or saccharin), or a flavoring agent (such as peppermint, methyl salicylate, or orange flavoring).
Injectable compositions are typically based on sterile injectable saline or phosphate buffered saline or other injectable carriers known in the art. As mentioned above, N-hydroxyamide derivatives of formula (I) are generally minor ingredients in such compositions, often in the range of 0.05-10% by weight, with the remainder being injectable carriers and the like.
The above ingredients for oral or injectable compositions are representative only. Further materials and processing techniques, etc., can be found in Remington's Pharmaceutical Sciences, fifth part of 20 th edition, 2000, Marck publishing company, Easton, Pennsylvania, which is incorporated herein by reference.
The compounds of the present invention may also be administered in sustained release form or from a sustained release drug delivery system. A description of representative sustained release materials is also found in the materials incorporated by Remington's pharmaceutical sciences.
Synthesis of the Compounds of the invention:
the novel derivatives of formula (I) can be obtained from readily available starting materials by several synthetic methods using solution phase and solid phase chemistry schemes. Examples of synthetic routes are described below.
The following abbreviations are respectively defined as follows:
aq (of water), eq (equivalent), h (hour), g (g), i.p. (intraperitoneal), L (L), mg (milligram), MHz (megahertz), min. (minute), mM (millimeter), μm (micrometer), mmol (millimole), mM (millimole), m.p. (melting point), mL (milliliter), μ L (microliter), p.o. (oral), s.c. (subcutaneous), BINAP (2, 2 '-bis (diphenylphosphino) -1, 1' -binaphthyl), CDCl (L)3(deuterated chloroform), CH3CN (acetonitrile), c-hex (cyclohexane), DCC (dicyclohexylcarbodiimide), DCM (dichloromethane), DIC (diisopropylcarbodiimide), DIEA (diisopropylethylamine), DMF (dimethylformamide), DMSO (dimethyl sulfoxide), DMSO-d6(deuterated dimethyl sulfoxide), EDC (1- (3-dimethyl-amino-propyl) -3-ethyl carbodiimide), ESI (electrospray ionization), Et2O (diethyl ether), HATU (dimethylamino- ([1, 2, 3))]Triazole [4, 5-b ]]Pyridin-3-yloxy) -methylene]-dimethyl-ammonium hexafluorophosphate), HPLC (high performance liquid chromatography), i-PrOH (2-propanol), LC (liquid chromatography), MeOH (methanol), MS (mass spectrometry), MTBE (methyl tert-butyl ether), NMM (N-methylmorpholine), NMR (nuclear magnetic resonance), RT (room temperature), PyBOP(benzotriazol-1-yl-oxy-tri-pyrrolidinyl-phosphino hexafluorophosphate), Rt (retention time), TBTU (2- (1-H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborateBoric acid), TEA (triethylamine), TFA (trifluoroacetic acid), THF (tetrahydrofuran), THP (tetrahydropyran), TLC (thin layer chromatography), UV (ultraviolet).
The synthesis method comprises the following steps:
a preferred method for preparing compounds of formula (I) is to couple a dioxolane protected dicarboxylic acid of formula (II) with a suitable amine (III) to form Intermediate (IV) wherein A, R1、R2、R4、R5、R6And R7As defined above (scheme 1 below). The general scheme for such coupling is given in the examples below, using conditions and methods well known to those skilled in the art, in a suitable solvent (e.g., DCM, THF or DMF), in the presence or absence of a base (e.g., TEA, DIEA, NMM), with or without the use of standard coupling reagents (e.g., DIC, EDC, TBTU, DCC, HATU, PyBOP)Isobutyl chloroformate, 1-methyl-2-chloropyridinium iodide (Mukaiyama's reagent), or others), to prepare amide bonds from amines and carboxylic acids or carboxylic acid derivatives (e.g., acid chlorides).
Scheme 1
The compounds of formula (III) may be obtained commercially or by the methods described herein.
The intermediates of formula (IV) can be reacted with hydroxylamines or with protected hydroxylamines H2NO-R8Reaction of wherein R8E.g., t-butyl, benzyl, trialkylsilyl or any suitable protecting group, followed by known deprotection steps to provide the compound of formula (I) (scheme 2, below).
Scheme 2
Intermediates of formula (II) may be prepared by known methods or by methods described herein.
An alternative route to the preparation of compounds of formula (I) may be with or without the use of standard coupling reagents (e.g. DIC, EDC, TBTU, DCC, HATU, PyBOP)Isobutyl chloroformate, 1-methyl-2-chloropyridinium iodide (Mukaiyama's reagent)), a carboxylic acid represented by the formula (V) is coupled with hydroxylamine or with protected hydroxylamine H2NO-R8Coupling of wherein R8E.g., t-butyl, benzyl, trialkylsilyl, tetrahydropyranyl or any suitable protecting group, followed by known deprotection steps to provide the compound of formula (I) (scheme 3 below).
Scheme 3
The HPLC data provided in the following examples were obtained by the following conditions: conditions A and B, HPLC column Waters XterraMS C8The column is 50mm multiplied by 4.6mm, and the flow rate is 2 mL/min; conditions C and D, Waters XterraMS C8The column was 150 mm. times.4.6 mm, and the flow rate was 1 mL/min.
Condition a: from 0.1% TFA in H2O to 0.07% TFA in CH3Gradient in CN for 8 min.
Condition B: from 95% H2O to 100% CH3Gradient CN 8 min.
Condition C: from 95% H2O to 100% CH3Gradient CN 20 min.
Condition D: from 95% H2O to 40% CH3Gradient CN 20 min.
All conditions were UV detected (maximum peak).
Preparative HPLC on Waters XterraPrep MS C810 μm column 300mm × 30mm, UV detection (254nM and 220nM), flow rate: 30 mL/min. The mass spectral data in the following examples were obtained as follows: mass spectrum: LC/MS Waters ZMD (ESI). The NMR data in the following examples were obtained as follows:1H-NMR：Bruker DPX-300MHz。
following further general procedures, compounds of formula (I) may be converted to other compounds of formula (I) using suitable conversion techniques well known to those skilled in the art.
If the general synthetic methods described above are not yet available to give compounds of formula (I) and/or intermediates required for the synthesis of compounds of formula (I), suitable preparative methods known to those skilled in the art should be used. In general, the synthetic route for each particular compound of formula (I) depends on the particular substituents per molecule and on whether the desired intermediate is readily available; these factors are also well known to those skilled in the art. All protection and deprotection methods are described in Kocienski, "Protecting Groups (Protecting Groups)", Georg Thieme Verlag Stuttgart, New York, 1994 and Greene and Wuts, "Protecting Groups in organic Synthesis (Protective Groups in organic Synthesis)", Wiley Interscience, 3 rd edition, 1999. It will be appreciated by those skilled in the art that certain reactions are preferably carried out in such a way that other potentially reactive functional groups of the molecule are masked or protected to avoid side reactions and/or to increase the yield of the reaction. Examples of protecting groups can be found in Kocienski (1994, supra) and Greene et al (1999, supra). The choice of protecting group and whether a protecting group is required for a particular reaction is well known to those skilled in the art and depends on the nature of the functional group to be protected (hydroxyl, amino, carboxyl, etc.), the structure and stability of the molecule of the moiety in which the substituent is in a reactive state.
The compounds of the invention associated with the solvent molecule may be isolated or purified by evaporative crystallization from a suitable solvent. Pharmaceutically acceptable acid addition salts of compounds of formula (I) containing a base centre may be prepared by conventional methods. For example, a solution of the free base can be treated with a suitable acid (either neat or dissolved in a suitable solvent) and the resulting salt isolated by filtration or evaporation of the reaction solvent in vacuo. Pharmaceutically acceptable base addition salts may likewise be obtained by treating a compound of formula (I) with a suitable base. The two types of salts may be formed or interconverted with ion exchange resins.
The present invention will now be illustrated by means of some examples, which should not be construed as limiting the scope of the invention.
The following are commercially available reagents and resins used：
2, 2-dimethoxypropane (ex Fluka), copper (II) chloride (ex Aldrich), HOBt (ex Aldrich), EDC (ex Aldrich), 1- (4-fluorophenyl) piperazine dihydrochloride (ex Aldrich), (R) - (-) -2-methylpiperazine (ex Astatech), 1-bromo-4- (trifluoromethoxy) benzene (ex Aldrich), 4-bromobiphenyl (ex Fluka), 2 '-bis (diphenylphosphino) -1, 1' -binaphthyl (ex Fluka).
Intermediate 1: (3R) -1-biphenyl-4-yl-3-methyl-piperazine
Toluene (700.00mL) was degassed with nitrogen for 30 minutes. (R) -2-methylpiperazine (30.0 g; 299.5 mmol; 1.0eq.) and 4-bromophenyl (73.3 g; 3)14.5 mmol; 1.05eq.), tBuONa (43.18 g; 449.3 mmol; 1.5eq.), palladium (II) acetate trimer (3.36 g; 15.0 mmol; 0.05eq.) and (+/-) -2, 2 '-bis (diphenylphosphino) -1, 1' -binaphthyl (7.46 g; 12mmol of the crude product; 0.04eq.) was added to the solution and the whole was heated under reflux overnight. The reaction mixture was filtered and Et was added2O into the filtrate precipitated the phosphine. The solvent was evaporated to dryness to afford a black solid (133 g). Purification by preparative chromatography (800g silica; DCM: MeOH 90: 10) afforded a black solid. The solid was poured into Et2To O, a minimum amount of DCM was added to completely dissolve it. Activated carbon was added and the resulting mixture was stirred at room temperature for 30 minutes. The celite layer was filtered and the solvent was evaporated until a white precipitate of off-white powder appeared. The mixture was cooled to-20 ℃ and filtered to obtain the product. The solid was treated with cold (0 ℃ C.) Et2O rinsing and drying at 45 ℃ under reduced pressure gave the first crop of the title compound (17.3g) as a white powder. The mother liquor was recrystallized to yield a second crop of white solid (13.8g, 41% overall yield). M+(ESI)：253.3.1H NMR(CDCl3300MHz) δ 7.47(d, J ═ 7.3Hz, 2H), 7.43(d, J ═ 8.7Hz, 2H), 7.32(t, J ═ 7.6Hz, 2H), 7.19(t, J ═ 8.2Hz, 1H), 6.91(d, J ═ 8.7Hz, 2H), 3.49(d, J ═ 11.9Hz, 2H), 2.89-3.06(m, 3H), 2.66(td, J ═ 11.4, 3.5Hz, 1H), 2.31(t, J ═ 10.6Hz, 1H), 1.06(d, J ═ 6.2Hz, 3H), HPLC (condition a): rt: 2.5min (HPLC purity: 98.5%).
Intermediate 2: (3R) -3-methyl-1- (4-trifluoromethoxyphenyl) -piperazine
Pd (OAc) under nitrogen atmosphere2(0.28g, 12.5mmol) and BINAP (0.62g, 1mmol) were added sequentially to a mixture of (R) -2-methylpiperazine (3.0g, 30mmol), 4-trifluoromethoxybromobenzene (6.6g, 27.5mmol) and sodium tert-butoxide (3.56g, 37.5mmol) in dry toluene (50mL) and refluxed for 16 hours. The reaction mixture was then concentrated and the crude product was purified by column chromatography on silica gel using methanol as eluent to afford the titled compoundCompound, as a dark brown solid (3g, 38%).
Example 1: (2R) -4- [4- (4-fluorophenyl) piperazin-1-yl]-N, 2-dihydroxy-4-oxobutanamide(1)
Step a) formation of (5R) -5- {2- [4- (4-fluorophenyl) piperazine 1-yl ] -2-oxyethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one
HOBt (2.97 g; 22.0 mmol; 1.1eq.) is added to [ (4R) -2, 2-dimethyl-5-oxo-1, 3-dioxolan-4-yl]A solution of acetic acid (3.48 g; 20.0 mmol; 1.0eq.) and TEA (6.07 g; 60.0 mmol; 3.0eq.) in DCM (60mL) was cooled to 0 ℃. EDC (4.6 g; 24.0 mmol; 1.2eq.) is then added and the resulting reaction mixture is stirred at 0 ℃ for 15 minutes. 1- (4-fluorophenyl) piperazine dihydrochloride (5.57 g; 22.0 mmol; 1.1eq.) was added and the resulting reaction mixture was stirred at room temperature overnight. Purification by flash chromatography (AcOEt/c-Hex: 50/50) gave the title compound as a colorless oil (5.12g, 76%). M+(ESI): HPLC (condition a): rt: 2.5min (HPLC purity: 97.4%).
Step b) formation of (2R) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide (1)
An aqueous hydroxylamine solution (50%, 0.295 mL; 5.0 mmol; 5.0eq.) was added to (5R) -5- {2- [4- (4-fluorophenyl) -1-piperazinyl]-2-oxyethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one (336 mg; 1.0 mmol; 1.0eq.) in a solution of i-PrOH/THF (25/75) (5 mL). After stirring at room temperature for 3 hours, the solvent was evaporated to give a solid. The solid was crystallized from AcOEt (Et added)2O and c-Hex) to yield the title compound as a white solid (25)0mg，80％)。M+(ESI)：312.1；M-(ESI)：310.1.1H NMR(DMSO-d6300MHz) δ 10.50(s, 1H), 8.72(s, 1H), 7.12-6.91(m, 4H), 5.46(d, J ═ 3.0Hz, 1H), 4.28(q, J ═ 6.3Hz, 1H), 3.60(s, 4H), 3.14-2.95(m, 4H), 2.65(d, J ═ 6.3Hz, 2H), HPLC (condition a): rt: 1.6min (HPLC purity: 85.6%).
Example 2: (2S) -4- [4- (4-fluorophenyl) piperazin-1-yl]-N, 2-dihydroxy-4-oxobutanamide (2)
Step a) formation of (5S) -5- {2- [4- (4-fluorophenyl) piperazin-1-yl ] -2-oxyethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one
The title product was prepared by the method of preparation example 1 (step a) but starting from [ (4S) -2, 2-dimethyl-5-oxo-1, 3-dioxolan-4-yl]Starting with acetic acid (300 mg; 1.72 mmol; 1.0eq.) the title compound was obtained as a white foam (350mg, 60%). M+(ESI)：337.1.1H NMR(CDCl3300MHz) δ 7.12-6.83(m, 4H), 4.94(dd, J ═ 3.0Hz, J ═ 7.5Hz, 1H), 3.90-3.68(m, 2H), 3.70-3.57(m, 2H), 3.19-3.08(m, 4H), 3.05(dd, J ═ 3.0Hz, J ═ 16.6Hz, 1H), 2.85(dd, J ═ 7.5Hz, J ═ 16.6Hz, 1H), 1.68(s, 3H), 1.63(s, 3H), HPLC (condition a): rt: 2.6min (HPLC purity: 96.9%).
Step b) formation of (2S) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide (2)
The title product was prepared according to the method for the preparation of example 1 (step b) but starting from (5S) -5- {2- [4- (4-fluorophenyl) piperazin-1-yl]-2-oxyethyl groupStarting from} -2, 2-dimethyl-1, 3-dioxolan-4-one (343mg, 1.02mmol), the title compound was obtained as a white powder (220mg, 69%). M+(ESI)：312.1；M-(ESI)：310.0.1H NMR(DMSO-d6300MHz) δ 10.53(s, 1H), 8.75(s, 1H), 7.18-6.85(m, 4H), 5.47(d, J ═ 3.0Hz, 1H), 4.28(q, J ═ 6.2Hz, 1H), 3.60(s, 4H), 3.16-2.93(m, 4H), 2.65(d, J ═ 6.3Hz, 2H), HPLC (condition a): rt: 1.2min (HPLC purity: 93.2%).
Example 3: (2S) -N, 2-dihydroxy-4- { (2R) -2-methyl-4- [4- (trifluoromethoxy) phenyl]Piperazine derivatives
Oxazin-1-yl } -4-oxobutanamide (3)
Step a) formation of (5S) -2, 2-dimethyl-5- [2- ((2R) -2-methyl-4- {4- [ (trifluoromethyl) oxy ] phenyl } piperazin-1-yl) -2-oxoethyl ] -1, 3-dioxolan-4-one
The title product was prepared by the method of preparation example 1 (step a) but starting from [ (4S) -2, 2-dimethyl-5-oxo-1, 3-dioxolan-4-yl]Acetic acid (150 mg; 0.86 mmol; 1.0eq.) and (3R) -3-methyl-1- {4- [ (trifluoromethyl) oxy-]Starting from phenyl } piperazine (intermediate 2, 247mg, 0.95mmol, 1.1eq.) the title compound was obtained as a colorless oil (123mg, 34%). M+(ESI)：417.2.1HNMR(DMSO-d6300MHz) δ 7.05(d, J ═ 8.3Hz, 2H), 6.79(d, J ═ 9.0Hz), 4.91-4.74(m, 1H), 4.51-4.39(m, 0.5H), 4.11-3.95(m, 0.5H), 3.68-3.21(m, 3H), 3.16-2.55(m, 4H), 1.57(s, 3H), 1.51(s, 3H), 1.40-1.20(m, 3H), PLC (condition a): rt: 4.3min (HPLC purity: 97.2%).
Step b) formation of (2S) -N, 2-dihydroxy-4- { (2R) -2-methyl-4- [4- (trifluoromethoxy) phenyl ] piperazin-1-yl } -4-oxobutanamide (3)
The title product was prepared according to the procedure for the preparation of example 1 (step b) but starting from (5S) -2, 2-dimethyl-5- [2- ((2R) -2-methyl-4- {4- [ (trifluoromethyl) oxy)]Phenyl } piperazin-1-yl) -2-oxoethyl]Starting from-1, 3-dioxolan-4-one (117mg, 0.28mmol), the title compound was obtained as a white powder (81mg, 74%). M+(ESI)：392.2；M-(ESI): 390.2 PLC (condition a): rt: 3.0min (HPLC purity: 93.8%).
Example 4: (2S) -4- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl]-N, 2-dihydroxy-4-
Oxybutylamide (4)
Step a) formation of (5S) -5- {2- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one
The title product was prepared by the method of preparation example 1 (step a) but starting from [ (4S) -2, 2-dimethyl-5-oxo-1, 3-dioxolan-4-yl]Starting from acetic acid (150 mg; 0.86 mmol; 1.0eq.) and (3R) -1-biphenyl-4-yl-3-methylpiperazine (intermediate 1, 239mg, 0.95mmol, 1.1eq.) the title compound was obtained as a colorless oil (107mg, 30%). M+(ESI): HPLC (condition a): rt: 4.3min (HPLC purity: 98.1%).
Step b) formation of (2S) -4- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide (4)
The title product was prepared according to the method for preparation example 1 (step b) but starting from (5S) -5- {2- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one (90mg, 0.22 mmol). The crude product was purified by reverse phase chromatography to give the title compound as a white powder (60mg, 71%). M + (ESI): 384.2, respectively; m- (ESI): HPLC (condition a): rt: 3.0min (HPLC purity: 99.0%).
Biological test:
the compounds of the present invention can be subjected to the following tests.
Example 5: enzyme inhibition assay
The compounds of the invention were tested for activity as MMP-1, MMP-2, MMP-9 and MMP-12 inhibitors.
MMP-9 assay protocol
Peptide substrate (7-methoxycoumarin-4-yl) acetyl-Pro-Leu-Gly-Leu- (3- [2, 4-dinitrophenyl) labeled with coumarin]-L-2, 3-diaminopropionyl) -Ala-Arg-NH2(McpaPLGLDpaAR) assay the inhibitory activity of the compounds of the invention on 92kDa gelatinase (MMP-9) was tested (Knight et al, 1992, FEBS Lett., 263-266).
The stock solution was prepared as follows: testing the cream: containing 100mM NaCl and 10mM CaCl2And 0.05% Brij35 in 100mM Tris-HCl pH 7.6.
Substrate: stock solutions (stored at-20 ℃) of 0.4mM McAPLGLDpaAR (from Bachem) (0.437mg/ml) were made up in 100% DMSO. Diluted to 8 μ M with test cream.
Enzyme: recombinant human 92kDa gelatinase (MMP-9; activated with APMA (phenylmercuric 4-aminoacetate) if necessary) was diluted appropriately with test milk fat.
A10 mM compound solution of test compound was prepared in 100% DMSO, diluted to 1mM with 100% DMSO, and then serially diluted 3-fold in 100% DMSO in each well of the row 1-10 of a 96-well microtiter plate. Test concentration range: 100mM (column 1) to 5.1nM (column 10).
The assay was performed in 96-well microtiter plates at a total volume of 100 μ L per well. Activated enzyme (20 μ L) was added to the wells followed by 20 μ L of test milk fat. Test compounds were added at appropriate concentrations dissolved in 10. mu.L DMSO, followed by 50. mu.L of McAPLGLDpaLR (8. mu.M, diluted in test milk fat with DMSO stock). The 10 concentrations of test compound were measured in duplicate at each concentration. Control wells contained neither enzyme nor test compound. The reaction was incubated at 37 ℃ for 2 hours. Without stopping the reaction, fluorescence at 405nm was immediately measured using a SLT Fluostar fluorimeter (SL T laboratories Gmb H, Grodig, Austria) with excitation at 320 nm.
The effect of the test compound was measured from a dose-response curve generated with 10 concentrations (in duplicate) of inhibitor. By fitting the data to the equation Y ═ a + ((b-a)/(1+ (c/X)d) To obtain IC)50(reduction of 50% of the enzyme activity required for the compound concentration). (Y-the inhibition obtained at the particular dose; X-the dose (nM); a-the minimum Y value or 0% inhibition; b-the maximum Y value or 100% inhibition; c-IC ═ the inhibition obtained at the particular dose50(ii) a d ═ slope). The result is rounded to a significant figure.
MMP-12 assay protocol
Peptide substrate (7-methoxycoumarin-4-yl) acetyl-Pro-Leu-Gly-Leu- (3- [2, 4-dinitrophenyl) labeled with coumarin]-L-2, 3-diaminopropionyl) -Ala-Arg-NH2(McpaPLGLDpaAR) assay the compounds of the invention were tested for their inhibitory activity against metalloelastase (MMP-12) (Knight et al, 1992, FEBS Lett., 263-266). This test protocol is as described above with respect to the MMP-9 assay.
MMP-1 assay protocol
Peptide substrate (7-methoxycoumarin-4-yl) acetyl-Pro-Leu-Gly-Leu- (3- [2, 4-dinitrobenzene) labelled with coumarin]-L-2, 3-diaminopropionyl) -Ala-Arg-NH2(McpaPLGLDpaAR) assay the inhibitory activity of the compounds of the invention on collagenase (MMP-1) was tested (Knight et al, 1992, FEBS Lett., 263-266). This test protocol is as above for MMP-9 description of the test method.
MMP-2 assay protocol
Peptide substrate (7-methoxycoumarin-4-yl) acetyl-Pro-Leu-Gly-Leu- (3- [2, 4-dinitrobenzene) labelled with coumarin]-L-2, 3-diaminopropionyl) -Ala-Arg-NH2(McpaPLGLDpaAR) assay the inhibitory activity of the compounds of the invention on gelatinase A (MMP-2) was tested (Knight et al, 1992, FEBS Lett., 263-266). This test protocol is as described above with respect to the MMP-9 assay.
IC for result50Expressed (concentration of compound required to reduce 50% of enzyme activity), IC of the compound of formula 150See Table 1 below
Table 1: IC for different MMPs50
| Examples | MMP-1IC50(nM) | MMP-12IC50(nM) |
| Example 1 | ＞5000 | 264 |
| Example 2 | ＞5000 | 58 |
| Example 3 | ＞5000 | 5 |
| Example 4 | ＞5000 | 5 |
Example 6: IL-2 induced peritoneal recruitment of lymphocytes
Intraperitoneal injection of IL-2 allows migration of lymphocytes into the peritoneal cavity. This is a model of lymphocyte migration occurring during the inflammatory phase
Scheme(s)
C3H/HEN mice (Elevige Janvier, France) were injected intraperitoneally with IL-2(Serono pharmaceutical Research Institute, 20. mu.g/kg, in saline).
The compounds of the invention were suspended in 0.5% carboxymethylcellulose (CMC)/0.25% Tween-20 and either injected subcutaneously or administered orally (10ml/kg) 15 minutes prior to IL-2 administration.
24 hours after IL-2 administration, peritoneal cavity was perfused three times with 5ml of Phosphate Buffered Saline (PBS) -1mM EDTA (+4 ℃) in succession and peritoneal leukocytes were collected. This suspension was centrifuged (1700 g.times.10 min. +4 ℃ C.) and the resulting pellet (pellet) was suspended in 1ml PBS-1mM EDTA.
Lymphocytes were identified and counted using a Beckman/Coulter counter.
Design of experiments
Animals were divided into 6 groups (6 mice per group):
group 1: (baseline) received 0.5% CMC/0.25% tween-20 (excipient for the compound of the invention) and physiological saline (excipient for IL-2);
group 2: (IL-2 control group) received 0.5% CMC/0.25% Tween-20 and injected with IL-2;
group 3: test group (inventive compound dose 1) received inventive compound and injected IL-2;
group 4: test group (inventive compound dose 2) received inventive compound and injected IL-2;
group 5: test group (inventive compound dose 3) received inventive compound and injected IL-2;
group 6: the reference group received the reference compound dexamethasone and injected IL-2.
Computing
Lymphocyte recruitment inhibition was calculated according to the following formula:
here, Ly 1 is the number of group 1 lymphocytes (E3/. mu.l), Ly2 is the number of group 2 lymphocytes (E3/. mu.l), and Ly X is the number of group X lymphocytes (3-5) (E3/. mu.l).
The results for the compounds of formula (I) are shown in Table 2 below.
Table 2: percent inhibition of IL-2-induced peritoneal recruitment of lymphocytes by Compounds of the invention
| Examples | Dosage (mg/kg) | Route of administration | Inhibition (%) |
| Example 2 | 1 | Is administered orally | 51 |
Example 7: model of Chronic Obstructive Pulmonary Disease (COPD)
This model allows the ability of the compounds of the invention to prevent smoking from causing COPD to be assessed.
Groups of 5 female AJ mice (Harlan, 17-25g) were placed in separate clean rooms and exposed to smoke (CS) daily for 11 consecutive days. Animals were weighed before treatment, on day 6 and day 12 of exposure. CS was produced using a 1R1 cigarette purchased from the tobacco institute at the university of Kentucky, USA and entered the chambers at a flow rate of 100 ml/min.
To minimize any potential problems caused by repeated exposure to high daily levels of CS, the CS exposure of mice from day 5 to day 11 was gradually increased over time to 6 cigarettes (exposure approximately 48 minutes).
Sham-exposed mice were also exposed to air for long periods of time daily as a control group (no exposure to CS).
Treatment of
The compounds of the invention were prepared using 0.5% sodium carboxymethylcellulose (CMC, Sigma reference C-4888) as excipient.
Animals were gavaged 2 times daily for a dose volume of 5ml/kg 1 hour before exposure to air or CS and 6 hours after cessation of exposure.
Sham-exposed animals (n-10) received vehicle daily and were exposed to air for up to 50 minutes. The control group (n-10) received vehicle and was exposed to CS (up to 6 cigarettes per day). The remaining groups were exposed to CS (up to 6 cigarettes per day) and treated with one of the test compounds or the reference compound.
Bronchoalveolar lavage and cytospin analysis
Bronchoalveolar lavage was performed 24 hours after the last CS exposure as follows:
the trachea was dissected under deep anesthesia (sodium pentobarbital) and inserted approximately 8mm with a Portex nylon intravenous cannula. The extract was gently and slowly infused 3 times with phosphate buffered saline (0.4ml) containing 10 units/ml heparin. Lavage fluid was placed into Eppendorf tubes and stored frozen prior to subsequent assays. The lavage fluid is then separated from the cells by centrifugation. The supernatant was removed and frozen for later analysis. The cell pellet was resuspended in PBS and the stained fraction (Turks staining) was counted under the microscope using a hemocytometer to calculate the total number of cells.
Different cell counts were then performed as follows: the remaining cell pellet was diluted to about 105 cells per ml. 500 μ l was placed in the funnel of a cytospin smear slide and centrifuged at 800rpm for 8 minutes. Slides were air dried and stained with Kwik-Diff solution (Shandon) according to commercial instructions. After the slides were dried, they were covered with coverslips and different cell counts were performed using an optical microscope. 400 cells were counted per slide. Cells were differentiated using standard morphometric techniques.
Statistical analysis
Mean +/-SD was calculated for each test group.
The analysis results were corrected using one-way analysis of variance (ANOVA) followed by Bonferroni with multiple point comparison. p < 0.05 was considered statistically significant.
Example 8: experimental Allergic Encephalomyelitis (EAE) model
The activity of the compounds of the invention can be evaluated in a mouse model of multiple sclerosis.
Animal(s) production
C57BL/6NCrlBR female mice were used. Mice were placed in wire mesh cages (cm 32 × 14 × 13h) with stainless steel feeders and fed standard feed (4RF21, Charles River, italy) with ad libitum access to water. Wet pellets (wet pellets) were also placed on the bottom of the cage each day, starting on day 7. Plastic bottles are used in addition to automatic water supply systems.
Experimental procedures
200 ug MOG in complete Freund's adjuvant (CFA, Difco, Detroit, USA) containing 0.5mg tubercle bacillus was injected subcutaneously in the left flank35-55Mice were immunized (day 0) with 0.2ml emulsion of peptide (Neosystem, Strasbourg, france). Immediately after immunization, 500ng of pertussis toxin (List Biological Lab., Campbell, CA, USA) in 400. mu.l buffer (0.5M NaCl, 0.017% Triton X-100, 0.015M Tris, pH 7.5) was intraperitoneally injected, and the animals were given a second injection of 500ng of pertussis toxin on day 2.
On day 7, mice were given a second subcutaneous injection of 200 μ g MOG in CFA to the right flank33-55A peptide. Starting approximately from day 8-10, this procedure resulted in progressive paralysis, starting from the tail and rising to the forelimbs.
Each animal was weighed and checked for paralysis according to the following scoring system score:
0 ═ no disease symptoms
0.5 ═ partial tail paralysis
Paralysis of the tail 1 ═ tail
1.5-Tail paralysis + partial unilateral posterior paralysis
Tail paralysis + bilateral hind limb weakness or partial paralysis
2.5 tail paralysis + partial hind limb paralysis (low pelvis)
3-tail paralysis + complete paralysis of hind limbs
3.5-Tail paralysis + hind limb paralysis + incontinence
4-tail paralysis + hind limb paralysis + weakness or partial forelimb paralysis
Dying or dead
Mortality and clinical signs were monitored daily for each group of animals by technicians unaware of the procedure.
All groups of animals were treated with compound, their vehicle or reference compound daily starting on day 7 for 15 or 21 consecutive days.
Histopathological examination
At the end of the treatment period, each animal was anesthetized with sodium pentobarbital and fixed by perfusion through the left ventricle using 4% paraformaldehyde as carotid artery. The fixed spinal cord is then carefully isolated.
Spinal cord sections were embedded with paraffin blocks. Sections and hematoxylin-eosin staining were performed, inflammation was checked with CD45 staining, demyelination and axonal loss were checked with Kluver-PAS (Luxol fast blue) plus iodic acid Schiff staining) and Bielchowski staining.
The total area of all spinal cord sections per animal was measured at 10X 10 grid intersections at a magnification of 0.4X 0.4mm per grid. Perivascular inflammatory infiltration was calculated in each section and expressed per mm2The number of infiltrations was evaluated to obtain a total value for each animal. The areas of demyelination and axonal loss for each animal were measured at 10X 10 grid intersections at a magnification of 0.1X 0.1mm per grid and expressed as a percentage of total demyelinated area to total area of the section.
Data evaluation and statistical analysis
Clinical and histopathological observations of each treatment group are presented as mean (+ -SEM). The values observed in the test drug-treated group were compared with the positive control group. Differences in clinical scores between groups were analyzed for significance using one-way ANOVA, followed by Fisher test hypothesis significance (p < 0.05).
Differences in perivascular inflammatory infiltrates and spinal cord demyelination, degree of axonal loss, and body weight data present between groups were analyzed by one-way ANOVA, followed by Fisher's test to hypothesize significance (p < 0.05).
Example 9: preparation of pharmaceutical dosage forms
The following dosage form examples illustrate representative pharmaceutical compositions, but the invention is not limited to these compositions.
Dosage form 1-tablet
The dry powder of the compound of the present invention is mixed with the dry gelatin binder in a mass ratio of about 1: 2. A small amount of magnesium stearate was added as a lubricant. The mixture was tabletted with a tabletting machine to tablets containing 80-90mg of the active N-hydroxyamide derivative per tablet with a tablet weight of 240-270 mg.
Dosage form 2-capsule
The dried compound of the present invention is mixed with a starch diluent in an approximate 1: 1 weight ratio. The mixture was filled into 250mg capsules each containing 125mg of the active N-hydroxyamide derivative.
Dosage form 3-liquid
The compound of the invention (1250mg), sucrose (1.75g) and xanthan gum (4mg) were mixed, passed through a 10 mesh US sieve and then mixed with previously prepared microcrystalline cellulose and sodium carboxymethylcellulose (11: 89, 50mg) in water. Sodium benzoate (10mg), fragrance and colour were diluted with water and added with stirring. Then a sufficient amount of water was added to make the total volume 5 ml.
Dosage form 4-tablet
The dry powder of the compound of the present invention is mixed with the dry gelatin binder in a weight ratio of about 1: 2. A small amount of magnesium stearate was added as a lubricant. This mixture was made into 450-900mg tablets (containing 150-300mg of the active N-hydroxyamide derivative) using a tablet press.
Formulation 5-injection
The compounds of the invention are dissolved in a sterile saline buffer medium for injection at a concentration of about 5 mg/ml.
Claims (12)
1. N-hydroxyamide derivatives shown in formula (I) or enantiomers, diastereomers, racemes or pharmaceutically acceptable salts thereof
Wherein:
a is N;
R1is phenyl, fluorophenyl, methoxyphenyl or biphenyl;
R2is H;
R3is H;
R4selected from H and C1-C6An alkyl group, a carboxyl group,
R5、R6and R7Is H.
2. The N-hydroxyamide derivative of claim 1 wherein R1Is phenyl; a is N.
3. The N-hydroxyamide derivative of claim 1 wherein said N-hydroxyamide derivative is selected from the group consisting of:
(2R) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
(2S) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
(2S) -N, 2-dihydroxy-4- { (2R) -2-methyl-4- [4- (trifluoromethoxy) phenyl ] piperazin-1-yl } -4-oxobutanamide; and
(2S) -4- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide.
4. Use of an N-hydroxyamide derivative according to any of claims 1-3 for the manufacture of a medicament for the treatment of a disease associated with modulation of matrix metalloprotease, wherein said matrix metalloprotease is selected from the group consisting of MMP-9, MMP-2 and MMP-12, said disease is associated with matrix metalloprotease activity and/or expression, and said disease associated with modulation of matrix metalloprotease is multiple sclerosis or chronic obstructive pulmonary disease.
5. A pharmaceutical composition comprising at least one N-hydroxyamide derivative according to any of claims 1-3 and a pharmaceutically acceptable carrier, diluent or excipient.
6. A process for the preparation of a compound of formula (I)
Comprising reacting a compound of formula (IV) with hydroxylamine H2NO-R8The reaction steps are as follows:
wherein
A is N;
R1is phenyl, fluorophenyl, methoxyphenyl or biphenyl;
R2is H;
R3is H;
R4selected from H and C1-C6An alkyl group, a carboxyl group,
R5、R6and R7Is a compound of formula (I) wherein the compound is H,
R8selected from the group consisting of H, t-butyl, benzyl, trialkylsilyl, and tetrahydropyranyl.
7. The method of claim 6, when R is8When not H, the method further comprises a deprotection step, wherein the deprotection step occurs at R8。
8. A compound of formula (IV) or a pharmaceutically acceptable salt thereof:
wherein
A is N;
R1is phenyl, fluorophenyl, methoxyphenyl or biphenyl;
R2is H;
R4selected from H and C1-C6An alkyl group, a carboxyl group,
R5、R6and R7Is H.
9. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
(5R) -5- {2- [4- (4-fluorophenyl) piperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one;
(5S) -5- {2- [4- (4-fluorophenyl) piperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one;
(5S) -2, 2-dimethyl-5- [2- ((2R) -2-methyl-4- {4- [ (trifluoromethyl) oxy ] phenyl } piperazin-1-yl) -2-oxoethyl ] -1, 3-dioxolan-4-one; and
(5S) -5- {2- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -2-oxoethyl } -2, 2-dimethyl-1, 3-dioxolan-4-one.
10. Use of a compound according to claim 1 in the manufacture of a medicament for inhibiting a matrix metalloproteinase selected from MMP-9, MMP-2 and MMP-12.
11. The use of claim 10, wherein R1Is phenyl; a is N.
12. The use according to claim 10, wherein the compound is selected from the group consisting of:
(2R) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
(2S) -4- [4- (4-fluorophenyl) piperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
(2S) -N, 2-dihydroxy-4- { (2R) -2-methyl-4- [4- (trifluoromethoxy) phenyl ] piperazin-1-yl } -4-oxobutanamide; and
(2S) -4- [ (2R) -4-biphenyl-4-yl-2-methylpiperazin-1-yl ] -N, 2-dihydroxy-4-oxobutanamide;
or a pharmaceutically acceptable salt thereof.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP05100646.8 | 2005-01-31 | ||
| EP05100646 | 2005-01-31 | ||
| US64893105P | 2005-02-01 | 2005-02-01 | |
| US60/648,931 | 2005-02-01 | ||
| PCT/EP2006/050480 WO2006079653A1 (en) | 2005-01-31 | 2006-01-27 | N-hydroxyamide derivatives and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1115132A1 HK1115132A1 (en) | 2008-11-21 |
| HK1115132B true HK1115132B (en) | 2012-07-27 |
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