Patent Grant
8927765
The present invention relates to compounds and their pharmaceutical uses. More particularly, the invention relates to substituted aromatic compounds, to processes for their preparation, to compositions comprising the same and to their use for the prevention or treatment of various diseases and conditions in subjects.
Blood Disorders
Hematopoiesis (hema=blood) refers to the process of formation, development and differentiation of all types of blood cells. All cellular blood components are derived from haematopoietic stem cells, including leukocytes and erythrocytes. The leukocytes or white blood cells (WBCs) are the cells of the immune system defending the body against both infectious disease and foreign materials. The erythrocytes are the non-nucleated, biconcave, disk-like cells which contain hemoglobin and these cells are essential for the transport of oxygen. A reduction in the number of white blood cells is called leukopenia whereas anemia refers to that condition which exists when there is a reduction below normal in the number of erythrocytes, the quantity of hemoglobin, or the volume of packed red blood cells in the blood. Disorders of the blood and the several kinds of leukopenia and anemia may be produced by a variety of underlying causes, including chemotherapy (e.g. chemotherapy induced anemia) and cancers (e.g. cancer related anemia). Therefore, there is a need for novel compositions and methods to stimulate hematopoiesis and to address the undesirable side effects of myelosuppression induced by chemotherapy and radiation therapy.
Kidney Diseases
The kidney is a structurally complex organ that has evolved to perform a number of important functions: excretion of the waste products of metabolism, regulation of body water and salt, maintenance of appropriate acid balance, and secretion of a variety of hormones and autocoids. Diseases of the kidney are as complex as its structure, but their study is facilitated by dividing them by their effects on four basic morphologic components: glomeruli, tubules, interstitium, and blood vessels. Unfortunately, some disorders affect more than one structure and the anatomic interdependence of structures in the kidney implies that damage to one almost always secondarily affects the others. Thus, whatever the origin, there is a tendency for all forms of renal disease ultimately to destroy all four components of the kidney, culminating in chronic renal failure. For instance, in autoimmune diseases such as diabetes mellitus, the kidneys are prime targets to suffer tissue damage or lesions. Nephrectomy, or kidney removal, a procedure which is sometimes performed on patients with kidney cancer (e.g. renal cell carcinoma), may negatively impact kidney function in the remaining kidney. Chemotherapy and immunosuppressive therapy are also a source of harmful effects to the kidneys. Therefore, there exists a need for drugs with a good safety profile which can be administered to patients with kidney disease. There is also a need for pharmaceutical compounds which can prolong kidney health or protect it from deterioration to the point at which the kidney can no longer function.
Inflammation
Immune Mediated inflammatory Disease (IMID) refers to any of a group of conditions or diseases that lack a definitive etiology but which are characterized by common inflammatory pathways leading to inflammation, and which may result from, or be triggered by, a dysregulation of the normal immune response. Autoimmune disease refers to any of a group of diseases or disorders in which tissue injury is associated with a humoral and/or cell-mediated immune response to body constituents or, in a broader sense, an immune response to self. Current treatments for autoimmune disease can be broadly classified into two groups: those drugs which dampen or suppress the immune response to self and those drugs which address the symptoms that arise from chronic inflammation. In greater detail, conventional treatments for autoimmune diseases (e.g., primarily arthritis) are (1) Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) such as aspirin, ibuprofen, naproxen, etodolac, and ketoprofen; (2) Corticosteroids such as prednisone and dexamethasone; (3) Disease-Modifying Anti-Rheumatic Drugs (DMARDs) such as methotrexate, azathioprine, cyclophosphamide, cyclosporin A. Sandimmune™, Neoral™, and FK506 (tacrolimus); (4) Biologicals such as the recombinant proteins Remicade™, Enbrel™ and Humira. While numerous therapies are available, conventional treatments are not routinely efficacious. More problematic is the accompanying toxicity which often prohibits the long-term use necessary with a chronic disease. Therefore, there is a need for compounds that are useful for the treatment of inflammatory-related diseases, including chronic and non-chronic autoimmune disease.
Oxidative Stress
Oxidative stress is caused by an imbalance between the production of reactive oxygen species and a biological system's ability to readily detoxify the reactive intermediates or easily repair the resulting damage. Although reactive oxygen species can be beneficial, as they are used in cell signaling and by the immune system they are also involved in many diseases. Therefore, a need still exists for compounds which can help maintain a proper balance in levels of reactive oxygen species in order to prevent damage to the cell or its components that may be caused by toxic effects of such reactive species.
The present invention addresses these needs for new treatment methods, compounds, and pharmaceutical compositions.
Indeed, it was unknown prior to the present invention that substituted aromatic compounds as defined herein may be therapeutically effective agents for the prevention and/or treatment of (i) blood disorders, (ii) renal disorders, nephropathy, and/or a renal disorder complication; (iii) an inflammatory-related disease; and/or (iv) oxidative stress related disorder.
Additional features of the invention will be apparent from review of the disclosure, figures and description of the invention below.
The present invention relates to compounds, compositions and treatment regimens for the prevention and/or treatment of various diseases and conditions in subjects.
Particular aspects of the invention relates to the use of compounds according to any of Formula I, Formula U. Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein, and pharmaceutically acceptable salts thereof. The salt may be selected from the group consisting of sodium, potassium, calcium and magnesium. Preferably, the compound is the sodium salt of Compound I or the sodium salt of compound XI. Specific examples of compounds according to the invention are represented in Table 2.
One particular aspect of the invention concerns the use of a compound represented by any of the formulas as defined herein for use in preventing and/or treating (i) blood disorders (e.g. anemia, neutropenia) (ii) renal disorders and/or renal disorder complications; (iii) inflammatory-related diseases (e.g. autoimmune disease); and (iv) oxidative stress.
Another related aspect of the invention concerns the use of a compound represented by any of the formulas as defined herein for the manufacture of a medications and pharmaceutical compositions. One particular example is a nephroprotective composition comprising a compound represented by any of the formulas as defined herein, and a pharmaceutically acceptable carrier.
The invention also relates to compounds according to any of Formula I, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula IIA as defined herein, and a pharmaceutically acceptable salts thereof, for use in preventing and/or treating (i) blood disorders (ii) renal disorders and/or renal disorder complications; (iii) inflammatory-related diseases; and/or (iv) oxidative stress.
The invention further relates to methods of preventing and/or treating various diseases and conditions including, but not limited (i) blood disorders (e.g. anemia, neutropenia) (ii) renal disorders and/or renal disorder complications; (iii) inflammatory-related diseases (e.g. autoimmune disease); and/or (iv) oxidative stress. The method comprises administering to a human patient in need thereof a pharmacologically effective amount of a compound represented by any of the formulas as defined herein.
The invention further relates to compounds according to any of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein, and pharmaceutically acceptable salts thereof, as prophylactically effective and/or therapeutically effective agents against various diseases and conditions in subjects.
Further aspects of the invention will be apparent to a person skilled in the art from the following description, and claims and generalizations therein.
In order that the invention may be readily understood, embodiments of the invention are illustrated by way of examples in the accompanying drawings.
A) General Overview of the Invention
The present inventors have discovered compounds that have beneficial pharmaceutical properties and that these compounds may be effective for use in the development of blood cells, in kidney protection, in inflammatory diseases, in diseases associated with high blood pressure and against oxidative stress-related disorders.
B) Compounds of the Invention
A compound of the present invention is represented by Formula I:
or a pharmaceutically acceptable salt thereof, wherein Cy, Q, and n are as defined hereinabove and hereinbelow.
The following are embodiments, groups, and substituents of the compounds according to Formula I, which are described hereinafter.
In one subset of compounds of Formula I, n is 1.
In another subset of compounds of Formula I, n is 0.
In one subset of compounds of Formula I, Cy is an aryl substituted with R1, R2, R3 and R4 as defined hereinabove and hereinbelow.
In another subset of compounds of Formula I, Cy is a heteroaryl substituted with R1, R2, R3, R4 and R5 as defined hereinabove and hereinbelow.
In one example, Cy is
wherein
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRcRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—;
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRcRd;
R4 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRcRd; and
R5 is H or ORb.
Rb, Rc and Rd are as defined hereinabove and hereinbelow.
In another example, Cy is
wherein
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRbRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRcRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—; and
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRbRd.
In one alternative example, Cy is
wherein
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRbRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRbRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C8 alkyl-Y—;
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRc; and
R4 is H.
In another alternative example, Cy is
wherein
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRcRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—;
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRc; and
R4 is H.
In another alternative example, Cy is
wherein
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRcRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—; and
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRc.
Thus compounds of Formula I comprise compounds of Formula II, III, IV and V:
wherein Q, n, R1, R2, R3, R4 and R5 are as defined hereinabove and hereinbelow.
Therefore, when n is 1, one subset of the compounds of Formula I comprise compounds of Formula IIA, IVA and VA:
wherein Q, R1, R2, R3, R4 and R5 are as defined hereinabove and hereinbelow.
Therefore, when n is 0, an alternative subset of the compounds of Formula I comprise compounds of Formula IIB, III, IV and VB:
wherein R1, R2, R3, R4 and R5 are as defined hereinabove and hereinbelow.
Thus, compounds of the present invention comprise compounds of Formula II:
or a pharmaceutically acceptable salt thereof;
n is an integer 0 or 1;
Q is C1-C4 alkyl optionally substituted with one Ra substituent;
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRcRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—;
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRcRd;
R4 is H, halogen, haloalkyl, C1-C4 alkyl, ORb. SRb, or NRcRd;
R5 is H or ORb;
Y is O, S, or NRcRd;
Ra is ORb, SRb, or NRcRd;
Rb is M, or C1-C4 alkyl; and
Rc and Rd are independently chosen from: H, or C1-C4 alkyl.
In one example, n is an integer 0 or 1.
In one example, when n is 1,
Q is C1-C4 alkyl optionally substituted with one Ra substituent;
R1 is C1-C6 alkyl, C2-C3 alkenyl, or C1-C6 alkyl-Y—, when R2 is H or halogen; or R1 is H, when R2 is C1-C6 alkyl;
R3 is H, ORb, SRb, or NRcRd;
R4 is H, ORb, SRb, or NRcRd;
R5 is H or ORb;
Y is O, S, or NRcRd; Ra is ORb, SRb, or NRcRd;
Rb is H, or C1-C4 alkyl; and
Rc and Rd are independently chosen from: H, or C1-C4 alkyl.
Thus, compounds of the present invention comprises compounds of Formula III:
or a pharmaceutically acceptable salt thereof,
wherein
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb. SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb. SRb, or NRcRd, when R2 is C1-C6 alkyl, C2/C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—;
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRcRd;
R4 is H, halogen, haloalkyl, C1-C4 alkyl, ORb. SRb, or NRcRd;
Y is O, S, or NRcRd;
Ra is ORb, SRb, or NRbRd;
Rb is H, or C1-C4 alkyl; and
Rc and Rd are independently chosen from: H, or C1-C4 alkyl.
In one example, R1 is C1-C6 alkyl, C2-C6 alkenyl, or C1-C6 alkyl-Y—, when R2 is H; or R1 is H, when R2 is C1-C6 alkyl, C2-C6 alkenyl, or C1-C6 alkyl-Y—;
R3 is H;
Y is O, S, or NRcRd; and
Rc and Rd are independently chosen from: H, or C1-C6 alkyl.
Thus, compounds of the present invention comprises compounds of Formula IV:
or a pharmaceutically acceptable salt thereof,
n is an integer 0 or 1;
Q is C1-C4 alkyl optionally substituted with one Ra substituent;
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb. SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb. SRb, or NRcRd, when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—;
R3 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRcRd;
R4 is H, halogen, haloalkyl, C1-C4 alkyl, ORb, SRb, or NRcRd;
Y is O, S, or NRcRd;
Ra is ORb, SRb, or NRcRd;
Rb is H, or C1-C4 alkyl; and
Rc and Rd are independently chosen from: H, or C1-C4 alkyl.
In one example, when n is 1. Q is C1-C4 alkyl;
R1 is C1-C6 alkyl-, when R2 is H; or R1 is H, when R2 is C1-C6 alkyl;
R3 is H; and
R4 is H.
Thus, compounds of the present invention comprises compounds of Formula V:
or a pharmaceutically acceptable salt thereof,
n is an integer 0 or 1;
Q is C1-C4 alkyl optionally substituted with one Ra substituent;
R1 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—, when R2 is H, halogen, haloalkyl, ORb, SRb, or NRcRd; or R1 is H, halogen, haloalkyl, ORb, SRb, or NRcRd when R2 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C1-C6 alkyl-Y—;
R4 is H;
Y is O, S, or NRcRa;
Ra is ORb, SRb, or NRcRa;
Rb is H, or C1-C4 alkyl; and
Rc and Rd are independently chosen from H, or C1-C4 alkyl.
In one example when n is 1, Q is C1-C4 alkyl;
R1 is C1-C6 alkyl, when R2 is H; or R1 is H, when R2 is C1-C6 alkyl; and
R4 is H.
In some embodiments, the compound according to Formula I excludes compounds where:
when n is 0, Cy is
and R1, R3, R4, and R5 are all H, then R2 cannot be ethyl, propyl, n-butyl or n-pentyl;
when n is 0, Cy is
and R2, R3, R4 and R5 are all H, then R1 cannot be ethyl, propyl or n-butyl;
when n is 0, Cy is
R1 is n-butyl, and R2, R4 and R5 are all H, then R3 cannot be Cl, Br or I; and/or
when n is 1, Q is CH2, Cy is
R1 is n-butyl, and R2, R4 and R5 are all H, then R3 cannot be Cl, Br, or I.
In some embodiments, the compound according to Formula I is limited to pharmaceutically acceptable salts and the acid form of the compound is excluded from the scope if the invention.
In some embodiments, the compounds of Table 2 are limited to pharmaceutically acceptable salts and the acid form of the compounds is excluded from the scope of the invention.
As used herein, the arrow → when used with Cy is intended to mean that Cy is covalently bonded to Q, when n is 1 or is covalently bonded to C(O)OH when n is 0.
As used herein, the term “alkyl” is intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, for example. C1-C6 as in C1-C6 alkyl is defined as including groups having 1, 2, 3, 4, 5, or 6, carbons in a linear or branched arrangement, or for example. C1-C4 as in C1-C4 alkyl is defined as including groups having 1, 2, 3, or 4 carbon atoms in a linear or branched arrangement. Examples of alkyl defined above include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, and i-butyl.
As used herein, the term, “alkenyl” is intended to mean unsaturated straight or branched chain hydrocarbon groups having the specified number of carbon atoms therein, and in which at least two of the carbon atoms are bonded to each other by a double bond, and having either E or Z regiochemistry and combinations thereof. For example, C2-C6 as in C2-C6 alkenyl is defined as including groups having 2, 3, 4, 5, or 6 carbons in a linear or branched arrangement, at least two of the carbon atoms being bonded together by a double bond. Examples of alkenyl include ethenyl (vinyl), 1-propenyl, 2-propenyl, 1-butenyl and the like as illustrated by compounds II and XI.
As used herein, the term “alkynyl” is intended to mean unsaturated, straight chain hydrocarbon groups having the specified number of carbon atoms therein and in which at least two carbon atoms are bonded together by a triple bond. For example C2-C6 as in C2-C6 alkynyl is defined as including groups having 2, 3, 4, 5, or 6 carbon atoms in a chain, at least two of the carbon atoms being bonded together by a triple bond. Examples of such alkynyls include ethynyl, 1-propynyl, 2-propynyl and the like.
As used herein, the term “halo” or “halogen” is intended to mean fluorine, chlorine, bromine and iodine.
As used herein, the term “haloalkyl” is intended to mean an alkyl as defined above, in which each hydrogen atom may be successively replaced by a halogen atom. Examples of haloalkyls include, but are not limited to, CH2F, CHF2 and CF3.
As used herein, the term “aryl”, either alone or in combination with another radical, means a carbocyclic aromatic monocyclic group containing 6 carbon atoms which may be further fused to a second 5- or 6-membered carbocyclic group which may be aromatic, saturated or unsaturated. Aryl includes, but is not limited to, phenyl and indanyl,
As used herein, the term “heteroaryl” is intended to mean an aromatic monocyclic ring system of up to six atoms, and contains from 1 to 4 hetero atoms selected from the group consisting of O, N, and S. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyrimidinyl, and pyrrolyl,
As used herein, the term “optionally substituted with one substituent” is intended to mean that the subsequently described event of circumstances may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. The definition is intended to mean from zero to one substituent.
If the substituents themselves are incompatible with the synthetic methods of the present invention, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions used in these methods. The protecting group may be removed at a suitable point in the reaction sequence of the method to provide a desired intermediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3.sup.rd ed.), John Wiley & Sons, NY (1999), which is incorporated herein by reference in its entirety. In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used in the methods of this invention. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful in an intermediate compound in the methods of this invention or is a desired substituent in a target compound.
The compounds of the present invention, or their pharmaceutically acceptable salts may contain one or more asymmetric centers, chiral axes and chiral planes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms and may be defined in terms of absolute stereochemistry, such as (R)- or (S)- or, as (O)- or (L)- for amino acids. The present invention is intended to include all such possible isomers, as well as, their racemic and optically pure forms. Optically active (+) and (−), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as reverse phase HPLC. The racemic mixtures may be prepared and thereafter separated into individual optical isomers or these optical isomers may be prepared by chiral synthesis. The enantiomers may be resolved by methods known to those skilled in the art, for example by formation of diastereoisomeric salts which may then be separated by crystallization, gas-liquid or liquid chromatography, selective reaction of one enantiomer with an enantiomer specific reagent. It will also be appreciated by those skilled in the art that where the desired enantiomer is converted into another chemical entity by a separation technique, an additional step is then required to form the desired enantiomeric form. Alternatively specific enantiomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents or by converting one enantiomer to another by asymmetric transformation.
Certain compounds of the present invention may exist in Zwitterionic form and the present invention includes Zwitterionic forms of these compounds and mixtures thereof.
Salts
As used herein, the term “pharmaceutically acceptable salt” is intended to mean base addition salts.
As used herein, the term “pharmaceutically acceptable base addition salt” is intended to mean those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine. N-ethylpiperidine, polyamine resins and the like.
Example of pharmaceutically acceptable salts are also described, for example, in Berge et al., “Pharmaceutical Salts”, J. Pharm. Sci. 66, 1-19 (1977).
Pharmaceutically acceptable salts may be synthesized from the parent agent that contains an acidic moiety, by conventional chemical methods. Generally, such salts are prepared by reacting the free acid forms of these agents with a stoichiometric amount of the appropriate base in water or in an organic solvent, or in a mixture of the two. Salts may be prepared in situ, during the final isolation or purification of the agent or by separately reacting a purified compound of the invention in its free acid form with the desired corresponding base, and isolating the salt thus formed.
All acid, salt and other ionic and non-ionic forms of the compounds described are included as compounds of the invention. For example, if a compound is shown as an acid herein, the salt forms of the compound are also included. Likewise, if a compound is shown as a salt and the acid forms are also included.
Prodrugs
In certain embodiments, the compounds of the present invention as represented by generalized Formula I, wherein said compounds are present in the free carboxylic acid form, may also include all pharmaceutically acceptable salts, isosteric equivalents such as tetrazole and prodrug forms thereof. Examples of the latter include the pharmaceutically acceptable esters or amides obtained upon reaction of alcohols or amines, including amino acids, with the free acids defined by Formula I.
Hydrates
In addition, the compounds of the invention also may exist in hydrated and anhydrous forms. Hydrates of any of the formulas described herein are included as compounds of the invention which may exist as a monohydrate or in the form of a polyhydrate.
C) Methods of Preparation
In general, all compounds of the present invention may be prepared by any conventional methods, using readily available and/or conventionally preparable starting materials, reagents and conventional synthesis procedures. Of particular interest is the work of Hundertmark, T.; Littke, A. F.; Buchwald, S. L.; Fu, G. C. Org. Lett. 2000, 12, pp. 1729-1731.
The inventors have discovered that a modified Sonogashira coupling can be used to synthesize compounds of the present invention. Generally speaking, Sonogashira coupling reactions may be represented as follows:
Typically, two catalysts are needed for this reaction: a zerovalent palladium complex and a halide salt of copper(I). The palladium complex activates the organic halides and the copper(I) halides react with the terminal alkyne and produce copper(I) acetylide, which acts as an activated species for the coupling reactions. The reaction medium must be basic to neutralize the hydrogen halide produced as the byproduct of this coupling reaction, so alkyl amine compounds such as triethylamine and diethylamine are sometimes used as solvents, but also DMF or ether can be used as solvent.
In this modified procedure, the inventors have used Pd(II) and eliminate the use of the second catalyst (copper(I) halides) and alkyl amine (triethylamine). This procedure offers the advantage of a practical route for scale-up of these compounds using a simple workup.
As used herein, the term “pharmaceutically acceptable carrier, diluent or excipient” is intended to mean, without limitation, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, emulsifier, or encapsulating agent, such as a liposome, cyclodextrins, encapsulating polymeric delivery systems or polyethyleneglycol matrix, which is acceptable for use in the subject, preferably humans.
D) Pharmaceutical Applications
As indicated hereinbefore and exemplified hereinafter, the compounds of the invention have beneficial pharmaceutical properties and these compounds may have useful pharmaceutical applications in the prevention and/or treatment of various diseases and conditions in a subject. Medical and pharmaceutical applications contemplated by the inventors include, but are not limited to, those addressing blood disorders, renal failure, inflammatory-related diseases and disorders related to reactive oxygen species.
The term “subject” includes living organisms in which blood disorders, renal failure, inflammatory-related diseases and/or oxidative stress-related disorders, can occur, or which are susceptible to such conditions. The term “subject” includes animals such as mammals or birds. Preferably, the subject is a mammal. More preferably, the subject is a human. Even more preferably, the subject is a human patient in need of treatment.
As used herein, “preventing” or “prevention” is intended to refer to at least the reduction of likelihood of the risk of (or susceptibility to) acquiring a disease or disorder (i.e., causing at least one of the clinical symptoms of the disease not to develop in a patient that may be exposed to or predisposed to the disease but does not yet experience or display symptoms of the disease). Biological and physiological parameters for identifying such patients are provided herein and are also well known by physicians.
The terms “treatment” or “treating” of a subject includes the application or administration of a compound of the invention to a subject (or application or administration of a compound of the invention to a cell or tissue from a subject) with the purpose of delaying, stabilizing, curing, healing, alleviating, relieving, altering, remedying, less worsening, ameliorating, improving, or affecting the disease or condition, the symptom of the disease or condition, or the risk of (or susceptibility to) the disease or condition. The term “treating” refers to any indication of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission; lessening of the rate of worsening; lessening severity of the disease; stabilization, diminishing of symptoms or making the injury, pathology or condition more tolerable to the subject; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; or improving a subject's physical or mental well-being. In some embodiments, the term “treating” can include increasing a subject's life expectancy and/or delay before additional treatments are required (e.g. dialysis or kidney transplantation).
Blood Disorders and Hematopoiesis
Addressing blood disorders is among the medical and pharmaceutical applications contemplated by present invention. The term “blood disorder” refers to any alteration in normal physiology, formation, proliferation and/or function of erythrocytes, leukocytes and/or platelets. Therefore, in one of its aspects the present invention relates to methods, compounds and compositions for stimulating hematopoiesis in a subject, preferably a human patient in need thereof.
Accordingly, one aspect of the invention relates to the use of the compounds described herein for stimulating production of leukocytes in a subject and/or for inhibiting decrease of leukocytes (i.e. leukopenia or leukocytopenia) in a subject. Related aspects include use of these compounds for stimulating a subject's immune system and reduce a subject's risk for infection. In some embodiment, the leukocytes are neutrophil granulocytes and the disorder is neutropenia. As is known, low white cell counts are often associated with chemotherapy, radiation therapy, leukemia, myelofibrosis and aplastic anemia. In addition, many common medications can cause leukopenia (eg. minocyclen, a commonly prescribed antibiotic). Accordingly, the invention also relates to the use of the compounds described herein for the prevention and/or treatment of those particular diseases and conditions.
In order to evaluate, assess, and/or confirm the efficacy of the method, compounds and/or compositions of the invention, serial measurements can be determined. Quantitative assessment of blood cell count, hennatopoiesis and erythropoiesis are well known in the art.
Typically a normal total white blood cell count in humans is within the range of 4 300 to 10 000 per mm3 (or mL), with an average value taken as 7 000 per mm3. A normal neutrophil count in human blood is within the range of 1 800 to 7 200 per mm3. Therefore, leukopenia refers to the condition wherein the blood white cell or leukocyte count is reduced to 5 000 per mm3 or less. In some embodiments, the subject is a human patient having a total white blood cells count under about 8 000 per mm3, or under about 5 000 per mm3 or under about 4 000 per mm3, or under 3 000 per mm3. In some embodiments, the subject is a human patient having a total neutrophil granulocytes count under about 5 000 per mm3, or under about 4 000 per mm3, or under about 3 000 per mm3, or under about 2 000 per mm3, or under about 1 000 per mm3. In some embodiments, the methods, compounds or compositions of the invention are effective in increasing the patients' total white blood cells count (and/or neutrophil granulocytes count) by at least 500 per mm3, by at least 1 000 per mm3, or by at least 2 000 per mm3 or more.
Another aspect of the invention relates to the use of the compounds described herein for stimulating production of erythrocytes (i.e. erythropoiesis) in a subject and/or inhibiting decrease of erythrocytes (i.e. anemia) in a subject. Related aspects include using of these compounds for compensating for excessive blood loss (e.g. a hemorrhage or chronically through low-volume loss), excessive blood cell destruction (e.g. hemolysis) or deficient red blood cell production (e.g. ineffective hematopoiesis). Related aspects include using of these compounds for blood cell differentiation, including the stimulation of production of erythrocytes from erythroid progenitor cells.
Of particular interest to the inventors is addressing anemia associated with the use of chemotherapy or radiotherapy in the treatment of cancer. Also of particular interest is anemia associated with end-stage renal disease as is the case for patients who require regular dialysis or kidney transplantation for survival. Therefore, some aspects of the invention relates to methods, compounds and compositions for the stimulation of the hematopoietic system in humans, for instance for treating the myelosuppressive effects of chemotherapy and/or radiotherapy and any other situation in which the stimulation of the hematopoietic system can be of therapeutic value such as, but not limited to, anemia. Additional aspects of the invention relates to a method effective for increasing the efficacy of chemotherapy and/or radiation therapy in human patients. The methods, compounds and compositions according to the invention may also be useful for using increasing the dose of chemotherapeutic compositions necessary to achieve a better therapeutic benefit, while avoiding increased side effects. Additional aspects relates to the methods, compounds and compositions according to the invention for reducing or eliminating chemotherapy-induced anemia in humans.
Typically, in normal adults, average values for red blood cell count (millions/mm3), hemoglobin (g/100 mL) and hematocrit or volume packed red blood cells (mL/100 mL) for females and males (at sea level) are 4.8+/−0.6 and 5.4+/−0.9, 14.0+/−2.0 and 16.0+/−2.0 and 52.0+/−5.0 and 47.0+/−5.0 respectively. Anemia refers to the condition which exists when there is a reduction below normal in the number of erythrocytes, the quantity of hemoglobin or the volume of packed red blood cells in the blood as characterized by a determination of the hematocrit. In some embodiments, the subject is a human patient having an hematocrit between 40 and 30, or under about 40. In some embodiments, the methods, compounds or compositions of the invention are effective in slowing a decrease or maintaining the patients' total red blood cells count and/or hematocrit. In some embodiments, the methods, compounds or compositions of the invention are effective in stabilizing the patients' hematocrit and/or in increasing the hematocrit by up about 5, or about 10, or whatever is necessary to achieve a normal value. In some embodiments, the methods, compounds or compositions of the invention are effective in reducing the need for blood transfusion(s).
Kidney Protection
In some aspects, the present invention relates to methods, compounds and compositions for preventing and/or treating a renal disorder in a subject in need thereof. The term “renal disorder”, “renal disease” or “kidney disease” means any alteration in normal physiology and function of the kidney. This can result from a wide range of acute and chronic conditions and events, including physical, chemical or biological injury, insult, trauma or disease, such as for example nephrectomy, chemotherapy, hypertension, diabetes, congestive heart failure, lupus, sickle cell anemia and various inflammatory, infectious and autoimmune diseases. HIV-associated nephropathies etc. This term includes but is not limited to diseases and conditions such as kidney transplant, nephropathy; chronic kidney disease (CKD); glomerulonephritis; inherited diseases such as polycystic kidney disease; nephromegaly (extreme hypertrophy of one or both kidneys); nephrotic syndrome; end stage renal disease (ESRD); acute and chronic renal failure; interstitial disease; nephritis; sclerosis, an induration or hardening of tissues and/or vessels resulting from causes that include, for example, inflammation due to disease or injury; renal fibrosis and scarring; renal-associated proliferative disorders; and other primary or secondary pathological conditions. Fibrosis associated with dialysis following kidney failure and catheter placement, e.g., peritoneal and vascular access fibrosis, is also included. In some embodiments the present invention more particularly relates to methods, compounds and compositions for nephroprotection. As used herein, “nephroprotection” refers to a process by which the rate of disease progression in the kidney is delayed or stopped and so the kidney is subsequently protected. In preferred embodiments (e.g. drug-induced nephrotoxicity), the compounds of Formula I are be administered prior to, during, or subsequent to the administration of a cytotoxic agent or anti-inflammatory or immunosuppressive drug. “Cytotoxic agent” refers to an agent which kills highly proliferating cells: e.g., tumors cells, virally infected cells, or hematopoietic cells. Examples of a cytotoxic agent include, but are not limited to, cyclophosphamide, doxorubicin, daunorubicin, vinblastine, vincristine, bleomycin, etoposide, topotecan, irinotecan, taxotere, taxol, 5-fluorouracil, methotrexate, gemcitabine, cisplatin, carboplatin, or chlorambucil, and an agonist of any of the above compounds. A cytotoxic agent can also be an antiviral agent: e.g., AZT (i.e., 3′-azido-3′-deoxythymidine) or 3TC/lamivudine (i.e., 3-thiacytidine). Such drugs can induce anemia in a mammal, including a human patient. In some embodiments, nephroprotection refers to the protection provided to a mammal from the toxic effects arising from treatment of the mammal with a chemotherapeutic agent. For instance, the compounds of Formula I may be used to protect the mammal, or facilitate its recovery, from the toxic effects resulting from treatment with a chemotherapeutic agent.
In some embodiments, the renal disorder or kidney disease may be generally defined as a “nephropathy” or “nephropathies”. The terms “nephropathy” or “nephropathies” encompass all clinical-pathological changes in the kidney which may result in kidney fibrosis and/or glomerular diseases (e.g. glomerulosclerosis, glomerulonephritis) and/or chronic renal insufficiency, and can cause end stage renal disease and/or renal failure. Some aspects of the present invention relate to compositions and their uses for the prevention and/or treatment of hypertensive nephropathy, diabetic nephropathy, and other types of nephropathy such as analgesic nephropathy, immune-mediated glomerulopathies (e.g. IgA nephropathy or Berger's disease, lupus nephritis), ischemic nephropathy. HIV-associated nephropathy, membranous nephropathy, glomerulonephritis, glomerulosclerosis, radiocontrast media-induced nephropathy, toxic nephropathy, analgesic-induced nephrotoxicity, cisplatin nephropathy, transplant nephropathy, and other forms of glomerular abnormality or injury; glomerular capillary injury (tubular fibrosis). In some embodiments, the terms “nephropathy” or “nephropathies” refers specifically to a disorder or disease where there is either the presence of proteins (i.e. proteinuria) in the urine of a subject and/or the presence of renal insufficiency.
The present invention further relates to methods, compounds and compositions for preventing and/or treating a renal disorder complication. The term “renal disorder complication” refers to a secondary condition correlated with a renal disorder, a health condition, an accident, or a negative reaction occurring during the course of a renal disorder that can become worse in its severity. A “renal disorder complication” is usually associated with increasing severity of the renal disease in the subjects suffering from symptoms or pathological changes, which can become widespread throughout the body or affecting other organ systems. As used herein, the term “renal disorder complication” encompasses, but is not limited to vascular diseases (e.g., macrovascular complications, microvascular complications, etc.), cardiovascular diseases (e.g. arteriosclerosis, atherosclerosis, coronary artery disease, congestive heart failure, stroke, angina, ischemic heat disease, myocardial infarction, etc), diabetic dyslipidemia, hyperlipidemia (e.g. hypercholesterolemia, hypertriglyceridemia, hyperlipoproteinennia), metabolic syndrome, obesity, anemia, edema, pancreatitis, weak bones, poor nutritional health and nerve damage.
According to some embodiments, the present invention concerns methods, compounds and compositions for preventing or treating characteristic aspects or evidence nephropathy including glomerulosclerosis, modification of the kidney vascular structure, and tubulointerstitial disease. Among characteristic aspects of nephropathy contemplated by the invention is the prevention of kidney cell apoptosis, fibrosis, sclerosis, and/or accumulation of proteins in tubular regions. Therefore, in some aspects the invention relates to a method for the prevention of kidney cell apoptosis, fibrosis, sclerosis, and/or accumulation of proteins in tubular regions. Related aspects concerns the use of the compounds and pharmaceutical compositions as defined herein for reducing CTGF mRNA expression and/or TGF-β mRNA expression in kidney cells.
In some embodiments, the subject may be suffering from a disorder such as, for example, diabetes, advanced progressive renal disease, and fibrotic renal disease and/or any of the renal diseases, renal disorders or renal disorder complications described herein. In some embodiments, the subject is a human patient having or susceptible of having glomerular filtration problems and/or a renal failure. In some embodiments, the subject is a human patient who is following, or who has received, treatments of chemotherapy or radiotherapy. Accordingly, related aspect concerns using the compound or pharmaceutical composition as defined herein for protecting kidneys against chemotherapeutic agents, including, but not limited to, doxorubicin, daunorubicin, vinblastine, vincristine, bleomycin, taxol, 5-fluorouracil, methotrexate, gemcitabine, cisplastin, carboplatin and chlorambucil. The methods of the present invention may comprise administering to a subject, e.g., a human patient in need thereof, a preventative- or therapeutically-effective amount of a compound or pharmaceutical composition as defined herein.
In order to evaluate, assess, and/or confirm the efficacy of the method, compounds and/or compositions of the invention, serial measurements can be determined. Quantitative assessment of renal function and parameters of renal dysfunction are well known in the art and can be found, for example, in Levey (Am J Kidney Dis. 1993, 22(I):207-214). Examples of assays for the determination of renal function/dysfunction are: serum creatinine level; creatinine clearance rate; cystatin C clearance rate, 24-hour urinary creatinine clearance, 24-hour urinary protein secretion; Glomerular Filtration Rate (GFR); urinary albumin creatinine ratio (ACR); albumin excretion rate (AER); and renal biopsy. Accordingly, in some aspects, the invention relates to a method of increasing creatinine clearance, to method of increasing insulin secretion and/or increasing insulin sensitivity, to method of decreasing insulin resistance by administering to a subject in need thereof a compound of Formula I.
In some embodiments, the subject is at risk of or has been diagnosed with, nephropathy. Typically a normal Glomerular Filtration Rate (GFR) in humans is from about 100 to about 140 ml/min. In some embodiments, the subject is a human patient having advanced nephropathy (i.e. a GFR of under 75 ml/min). In some embodiments, the subject is a human patient having ESRD (i.e. GFR of less than 10 ml/min). In some embodiments, the methods, compounds or compositions of the invention are effective in increasing the patients' GFR value by at least 1, 5, 10, 15, 20 or 25 ml/min or more.
In some embodiments, the subject is at risk of or has been diagnosed with, a kidney disease. In various embodiments, the subject is a human patient having or progressing towards stage I kidney disease, stage II kidney disease, stage III kidney disease, stage 1V kidney disease or stage V kidney disease. In some embodiments, the methods, compounds or compositions of the invention are effective in stabilizing or in improving the patient's kidney disease ((e.g. from stage V to stage IV, or from stage IV to stage III, or from stage III to stage II, or from stage II to stage I).
One of the first clinical indications of nephropathy is the presence of albuminuria or proteinuria. One refers to microalbuminuria when the amount of albumin in the urine is less than or equal to <300 mg/day and proteinuria when the total amount of protein in the urine is greater than 1 g/day. According to some aspects, the invention related to a method of preventing or decreasing proteinuria by administering to a subject in need thereof a compound of Formula I. In some embodiments, the subject is at risk of, or has been diagnosed with, proteinuria. In some embodiments, the subject is a human patient producing less than about 300 mg/day of protein in its urine. In some embodiments, the subject is a human patient producing more than about 1 g/day of protein in its urine. In some embodiments, the subject is a human patient having microalbuminuria. In some embodiments, the subject is a human patient with an albumin amount in the urine that exceeds 200 μg/min. In some embodiments, the methods, compounds or compositions of the invention are effective in lowering the patient's albuminuria by at least 10, 25, 50, 75, 100, 150, 200 μg/min or more.
Effectiveness of the methods, compounds and compositions of the invention may be assessed by the reduction in the undesired symptoms. Such reduction may be determined for example by the improvement in renal function as compared to the function prior to treatment. Such remediation may be evident in a delay in the onset of renal failure (including dialysis or transplant) or in a decrease in the rate of the deterioration of renal function as determined for example by the slowing of the rate of the increase of proteinuria or slowing the rate of the rise in serum creatinine or by the fall in the parameter of creatinine clearance or GFR, or decrease in hospitalization rate or mortality. In preferred embodiments, the compound is Compound I or Compound XI, or a pharmaceutically acceptable salt thereof.
In one embodiment, a compound of the invention is used in combination with at least one additional known compound which is currently being used or in development for preventing or treating renal disorder such as nephropathy, or an associated disorder or complication. Examples of such known compounds include but are not limited to: ACE inhibitor drugs (e.g. captopril (Capoten®), enalapril (Innovace®), fosinopril (Staril®), lisinopril (Zestril®), perindopril (Coversyl®), quinapril (Accupro®), trandanalopril (Gopten®), lotensin, moexipril, ramipril); RAS blockers; angiotensin receptor blockers (ARBs) (e.g. Olmesartan, Irbesartan, Losartan, Valsartan, candesartan, eprosartan, telmisartan, etc); protein kinase C (PKC) inhibitors (e.g. ruboxistaurin); inhibitors of AGE-dependent pathways (e.g. aminoguanidine, ALT-946, pyrodoxamine (pyrododorin), OPB-9295, alagebrium); anti-inflammatory agents (e.g. clyclooxigenase-2 inhibitors, mycophenolate mophetil, mizoribine, pentoxifylline), GAGs (e.g. sulodexide (U.S. Pat. No. 5,496,807)); pyridoxamine (U.S. Pat. No. 7,030,146); endothelin antagonists (e.g. SPP 301), COX-2 inhibitors, PPAR-γ antagonists and other compounds like amifostine (used for cisplatin nephropathy), captopril (used for diabetic nephropathy), cyclophosphamide (used for idiopathic membranous nephropathy), sodium thiosulfate (used for cisplatin nephropathy), tranilast, etc.
Inflammation
Another aspect of the invention relates to the use of the compounds of the invention for the prevention and/or treatment of inflammatory-related diseases. The term “inflammatory-related disease” refers to any and all abnormalities associated with inflammation, including chronic and acute inflammatory diseases, including but not limited to immune mediated inflammatory diseases (IMID) and autoimmune diseases arthritis, ITP, glomerulonephritis, vasculitis, psoriatic arthritis, systemic lupus erythematoses (SLE), idiopathic thrombocytopenic purpura (ITP), psoriasis, Crohn's disease, inflammatory bowel disease, ankylosing spondylitis, Sjögren's syndrome, Still's disease (macrophage activation syndrome), uveitis, scleroderma, myositis, Reiter's syndrome, and Wegener's syndrome. In general, prophylactic and therapeutic uses comprise the administration of a compound as described herein to a subject, preferably a human patient in need thereof. The compounds of the invention may be administered with any conventional treatments, including more particularly the current treatments defined hereinbefore in the Background section. In order to evaluate, assess, and/or confirm the efficacy of the method, compounds and/or compositions of the invention, serial measurements can be determined. Quantitative methods and techniques for the assessment of inflammation are well known in the art and include for instance methods similar to those provided in the exemplification section.
Additionally, or alternatively, compounds of the present invention may inhibit the inhibiting the production of prostaglandins, including but not limited to PGE2, and thus be useful to reduce fever, pain, stiffness, and swelling.
Oxidative Stress
Another aspect of the invention relates to methods, compounds and compositions of the invention for the prevention and/or treatment of an oxidative stress related disorder. The term “oxidative stress related disorder” refers to any disease in which there is an imbalance between the production of reactive oxygen species and a biological system's ability to readily detoxify the reactive intermediates or easily repair the resulting damage. Examples of such diseases include, but are not limited to, cardiovascular diseases, cancer, diabetes, arthritis, atherosclerosis. Parkinson's disease, heart failure, myocardial infarction. Alzheimer's disease, chronic fatigue syndrome and autoimmune diseases.
In general, prophylactic and therapeutic uses comprise the administration of a compound as described herein to a subject, preferably a human patient in need thereof. In some embodiments, the subject is at risk of, or has been diagnosed with an oxidative stress related disorder as defined hereinabove.
A related aspect of the invention relates to methods, compounds and compositions for maintaining a proper balance in levels of reactive oxygen species, and more particularly nitric oxide (NO), in order to prevent damage to the cell or its components. An additional related aspect relates to methods, compounds and compositions for preventing damage to a cell or its components (including but not limited to proteins, lipids and DNA) that may be caused by reactive species, and more particularly NO. Yet, a further aspect of the invention relates to the use of methods, compounds and compositions according to the invention for inhibiting NO production and/or for inhibiting the enzyme nitric oxide synthase. These methods comprise contacting the cell, component or enzyme with a compound and/ora composition as defined herein. Quantitative methods and techniques for the assessment of reactive oxygen species levels in vitro and in vivo are well known in the art.
In general, prophylactic and therapeutic uses comprise the administration of a compound as described herein to a subject, preferably a human patient in need thereof. The compounds of the invention may be administered with various antioxidants including, but not limited to, metal chelators/scavengers (e.g. desferrioxamine [Desferal®], a low molecular weight substance capable to scavenge Fe3+ and other metal ions); small scavengers of .O2− (superoxide), .OH (hydroxyl) or NO (nitric oxide) radicals (e.g. acetyl salicylic acid, scavenger of .O2−; mannitol or captopril, scavengers of .OH; arginine derivatives, inhibitors of nitric oxide synthase which produce NO); and proteins or their fragments that can assist the protective action against reactive oxygen species (e.g. superoxide dismutase which breaks down .O2−; hemoglobin which traps NO, catalase, or glutathione peroxidase which can eliminate hydrogen peroxide).
E) Pharmaceutical Compositions and Formulations
A related aspect of the invention concerns pharmaceutical compositions comprising one or more of the compounds of the invention described herein. As indicated hereinbefore, the compounds of the invention may be useful in: (i) in preventing and/or treating blood disorders (e.g. by stimulating hematopoiesis); (ii) in preventing, and/or treating a renal disorder, a nephropathy, and/or a renal disorder complication; (iii) in preventing and/or treating a inflammatory-related disease (e.g. an autoimmune disease); and/or (iv) in the prevention and/or treatment of an oxidative stress related disorder.
As used herein, the term “therapeutically effective amount” means the amount of compound that, when administered to a subject for treating or preventing a particular disorder, disease or condition, is sufficient to effect such treatment or prevention of that disorder, disease or condition. Dosages and therapeutically effective amounts may vary for example, depending upon a variety of factors including the activity of the specific agent employed, the age, body weight, general health, gender, and diet of the subject, the time of administration, the route of administration, the rate of excretion, and any drug combination, if applicable, the effect which the practitioner desires the compound to have upon the subject and the properties of the compounds (e.g. bioavailability, stability, potency, toxicity, etc), and the particular disorder(s) the subject is suffering from. In addition, the therapeutically effective amount may depend on the subject's blood parameters (e.g. lipid profile, insulin levels, glycemia), the severity of the disease state, organ function, or underlying disease or complications. Such appropriate doses may be determined using any available assays including the assays described herein. When one or more of the compounds of the invention is to be administered to humans, a physician may for example, prescribe a relatively low dose at first, subsequently increasing the dose until an appropriate response is obtained.
As used herein, the term “pharmaceutical composition” refers to the presence of at least one compound of the invention according to any one of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein and at least one pharmaceutically acceptable vehicle. Examples of representative compounds of the invention include the compounds in Table 2 and pharmaceutically acceptable salts thereof.
“Pharmaceutically acceptable vehicle” refers to a diluent, adjuvant, excipient, or carrier with which a compound is administered. The term “pharmaceutically acceptable” refers to drugs, medicaments, inert ingredients etc., which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, incompatibility, instability, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. It preferably refers to a compound or composition that is approved or approvable by a regulatory agency of the Federal or State government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals and more particularly in humans. The pharmaceutically acceptable vehicle can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils. Additional examples of pharmaceutically acceptable vehicles include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate. Prevention of the action of microorganisms can be achieved by addition of antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, isotonic agents are included, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, in the composition. Prolonged absorption of injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.
In some embodiments, the compositions of the invention comprise an effective amount of a compound of Formula I and, more preferably, Formula II as described herein before. Particularly preferred are compounds I, II, IV, VII, X, XI, and XIII. More preferred are the sodium salts of 3-pentylphenylacetic acid, 3-hydroxy-5-pentylphenylacetic acid and 3-hexylbenzoic acid.
In some embodiments the invention pertains to pharmaceutical compositions for preventing and/or treating blood disorders that include one or more compounds of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein.
In some embodiments the invention pertains to pharmaceutical compositions for preventing and/or treating a renal disorder, a nephropathy, and or a renal disorder complication, the composition comprising one or more compounds of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein.
In some embodiments the invention pertains to pharmaceutical compositions for preventing and/or treating a inflammatory-related disease, the composition comprising one or more compounds of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein.
In some embodiments the invention pertains to pharmaceutical compositions for preventing, delaying and/or treating an oxidative stress related disorder, the composition comprising one or more compounds of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein.
The compounds of the invention may be formulated prior to administration into pharmaceutical compositions using available techniques and procedures. For instance, the pharmaceutical compositions may be formulated in a manner suitable for administration by oral, intravenous (iv), intramuscular (IM), depo-im, subcutaneous (sc), depo-sc, sublingually, intranasal, intrathecal topical or rectal routes.
Preferably, the compound(s) of the invention can be orally administered. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with a pharmaceutically acceptable vehicle (e.g. an inert diluent or an assimilable edible carrier) and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product. The amount of the therapeutic agent in such therapeutically useful compositions is such that a suitable dosage will be obtained.
Formulations of the invention suitable for oral administration may be in the form of capsules (e.g. hard or soft shell gelatin capsule), cachets, pills, tablets, lozenges, powders, granules, pellets, dragees, e.g., coated (e.g., enteric coated) or uncoated, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. A compound of the present invention may also be administered as a bolus, electuary or paste, or incorporated directly into the subject's diet. Moreover, in certain embodiments these pellets can be formulated to (a) provide for instant or rapid drug release (i.e., have no coating on them); (b) be coated, e.g., to provide for sustained drug release over time; or (c) be coated with an enteric coating for better gastrointestinal tolerability. Coating may be achieved by conventional methods, typically with pH or time-dependent coatings, such that the compound(s) of the invention is released in the vicinity of the desired location, or at various times to extend the desired action. Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, waxes, and shellac.
In solid dosage forms for oral administration a compound of the present invention may be mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; absorbents, such as kaolin and bentonite clay; lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
Peroral compositions typically include liquid solutions, emulsions, suspensions, and the like. The pharmaceutically acceptable vehicles suitable for preparation of such compositions are well known in the art. Typical components of carriers for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. For a suspension, typical suspending agents include methyl cellulose, sodium carboxymethyl cellulose, tragacanth, and sodium alginate; typical wetting agents include lecithin and polysorbate 80; and typical preservatives include methyl paraben and sodium benzoate. Peroral liquid compositions may also contain one or more components such as sweeteners, flavoring agents and colorants disclosed above.
Pharmaceutical compositions suitable for injectable use may include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. Sterile injectable solutions can be prepared by incorporating the therapeutic agent in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the therapeutic agent into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient (i.e., the therapeutic agent) plus any additional desired ingredient from a previously sterile-filtered solution thereof.
Pharmaceutical formulations are also provided which are suitable for administration as an aerosol, by inhalation. These formulations comprise a solution or suspension of the desired compound of any Formula herein or a plurality of solid particles of such compound(s). For instance, metal salts of the compounds of this invention are expected to have physical chemical properties amenable with the preparation of fine particles of active pharmaceutical ingredient (API) for administration by inhalation but not the free acid form of these compounds. The desired formulation may be placed in a small chamber and nebulized. Nebulization may be accomplished by compressed air or by ultrasonic energy to form a plurality of liquid droplets or solid particles comprising the agents or salts. The liquid droplets or solid particles should have a particle size in the range of about 0.5 to about 5 microns. The solid particles can be obtained by processing the solid agent of any Formula described herein, or a salt thereof, in any appropriate manner known in the art, such as by micronization. The size of the solid particles or droplets will be, for example, from about 1 to about 2 microns. In this respect, commercial nebulizers are available to achieve this purpose. A pharmaceutical formulation suitable for administration as an aerosol may be in the form of a liquid, the formulation will comprise a water-soluble agent of any Formula described herein, or a salt thereof, in a carrier which comprises water. A surfactant may be present which lowers the surface tension of the formulation sufficiently to result in the formation of droplets within the desired size range when subjected to nebulization.
The compositions of this invention may also be administered topically to a subject, e.g., by the direct laying on or spreading of the composition on the epidermal or epithelial tissue of the subject, or transdermally via a “patch”. Such compositions include, for example, lotions, creams, solutions, gels and solids. These topical compositions may comprise an effective amount, usually at least about 0.1%, or even from about 1% to about 5%, of a compound of the invention. Suitable carriers for topical administration typically remain in place on the skin as a continuous film, and resist being removed by perspiration or immersion in water. Generally, the carrier is organic in nature and capable of having dispersed or dissolved therein the therapeutic agent. The carrier may include pharmaceutically acceptable emollients, emulsifiers, thickening agents, solvents and the like.
Other compositions useful for attaining systemic delivery of the subject agents may include sublingual, buccal and nasal dosage forms. Such compositions typically comprise one or more of soluble filler substances such as sucrose, sorbitol and mannitol; and binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose. Glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.
The compound(s) of the invention may also be administered parenterally, intraperitoneally, intraspinally, or intracerebrally. For such compositions, the compound(s) of the invention can be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
The method of treatment of the present invention may also include co-administration of the at least one compound according to the invention, or a pharmaceutically acceptable salt thereof together with the administration of another therapeutically effective agent for the prevention and/or treatment of (i) blood disorders, (ii) renal disorder, a nephropathy, and/or a renal disorder complication; (iii) an inflammatory-related disease; and/or (iv) oxidative stress related disorder. Therefore, an additional aspect of the invention relates to methods of concomitant therapeutic treatment of a subject, comprising administering to a subject in need thereof an effective amount of a first agent and a second agent, wherein the first agent is as defined in Formula I, and the second agent is for the prevention or treatment of any one of disorder or disease of (i) to (v) hereinbefore. As used herein, the term “concomitant” or “concomitantly” as in the phrases “concomitant therapeutic treatment” or “concomitantly with” includes administering a first agent in the present of a second agent. A concomitant therapeutic treatment method includes methods in which the first, second, third or additional agents are co-administered. A concomitant therapeutic treatment method also includes methods in which the first or additional agents are administered in the presence of a second or additional agents, wherein the second or additional agents, for example, may have been previously administered. A concomitant therapeutic treatment method may be executed step-wise by different actors. For example, one actor may administer to a subject a first agent and as a second actor may administer to the subject a second agent and the administering steps may be executed at the same time, or nearly the same time, or at distant times, so long as the first agent (and/or additional agents) are after administration in the presence of the second agent (and/or additional agents). The actor and the subject may be the same entity (e.g. a human).
Accordingly, the invention also relates to a method for preventing, reducing or eliminating a symptom or complication of any one of the above mentioned disease or condition. The method comprises administering, to a subject in need thereof, a first pharmaceutical composition comprising at least one compound of the invention and a second pharmaceutical composition comprising one or more additional active ingredients, wherein all active ingredients are administered in an amount sufficient to inhibit, reduce, or eliminate one or more symptoms or complications of the disease or condition to be treated. In one aspect, the administration of the first and second pharmaceutical composition is temporally spaced apart by at least about two minutes. Preferably the first agent is a compound of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein, or a pharmaceutically acceptable salt thereof, e.g. sodium salt. The second agent may be selected from the list of compounds given hereinbefore.
F) Kits
The compound(s) of the invention may be packaged as part of a kit, optionally including a container (e.g. packaging, a box, a vial, etc). The kit may be commercially used according to the methods described herein and may include instructions for use in a method of the invention. Additional kit components may include acids, bases, buffering agents, inorganic salts, solvents, antioxidants, preservatives, or metal chelators. The additional kit components are present as pure compositions, or as aqueous or organic solutions that incorporate one or more additional kit components. Any or all of the kit components optionally further comprise buffers.
The compound(s) of the invention may or may not be administered to a patient at the same time or by the same route of administration. Therefore, the methods of the invention encompass kits which, when used by the medical practitioner, can simplify the administration of appropriate amounts of two or more active ingredients to a patient.
A typical kit of the invention comprises a unit dosage form of a at least one compound according to the invention, e.g., a compound of Formula I, Formula II, Formula IIA, Formula IIB, Formula III, Formula IV, Formula IVA, Formula IVB, Formula V, Formula VA as defined herein or a pharmaceutically acceptable salt thereof, and a unit dosage form of at least one additional active ingredient. Examples of additional active ingredients that may be used in conjunction with the compounds according to the invention, include, but are not limited to any of the compounds that could be used in combination with the compound(s) of the invention as indicated herein before.
Kits of the invention can further comprise devices that are used to administer the active ingredients. Examples of such devices include, but are not limited to, syringes, drip bags, patches, inhalers, enemas, and dispensers for the administration of suppository formulations.
Kits of the invention can further comprise pharmaceutically acceptable vehicles that can be used to administer one or more active ingredients. For example, if an active ingredient is provided in a solid form that must be reconstituted for parenteral administration, the kit can comprise a sealed container of a suitable vehicle in which the active ingredient can be dissolved to form a particulate-free sterile solution that is suitable for parenteral administration. Examples of pharmaceutically acceptable vehicles are provided hereinbefore.
Headings are included herein for reference and to aid in locating certain sections These headings are not intended to limit the scope of the concepts described therein under, and these concepts may have applicability in other sections throughout the entire specification Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
The singular forms “a”, “an” and “the” include corresponding plural references unless the context clearly dictates otherwise.
Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, concentrations, properties, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the present specification and attached claims are approximations that may vary depending upon the properties sought to be obtained. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the embodiments are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contain certain errors resulting from variations in experiments, testing measurements, statistical analyses and such.
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of this invention and covered by the claims appended hereto. The invention is further illustrated by the following examples, which should not be construed as further limiting.
The Examples set forth herein below provide exemplary methods for the preparation of certain representative compounds encompassed by general Formula I. Some Examples provide exemplary uses of certain representative compounds of the invention. Also provided are exemplary methods for assaying the compounds of the invention for in vitro and in vivo efficacy.
Instrumentation:
All HPLC chromatograms and mass spectra were recorded on an HP 1100 LC-MS Agilent instrument using an analytical C18 column (250×4.6 mm, 5 microns) with a gradient over 5 min of 15-99% CH3CN—H2O with 0.01% TFA as the eluant and a flow of 2 mL/min.
Compound I: Synthesis using Modified Procedure of Sonogashira:
Step 1:
To a solution/suspension of 3-bromophenylacetic acid (5.02 g, 23.33 mmol) in ethanol (100 mL) at room temperature was added concentrated sulfuric acid (1 mL). The colorless solid was then stirred overnight at 80° C. The solution was concentrated under reduced pressure. The residue was diluted with ethyl acetate (25 mL), water (25 mL) and the two layers were separated. The aqueous layer was extracted with 2× ethyl acetate (25 mL) and brine (20 mL). The combinated organic layers were washed with 2× saturated solution of NaHCO3 (25 mL), brine (25 mL) and dried over sodium sulfate. After filtration the solution it was evaporated to dryness. This gave a light yellow oil (5.4 g, 95%). 1H-NMR (400 MHz, CDCl3): δ 1.26 (t, J=4.7 Hz, 3H), 3.57 (s, 2H), 4.15 (Q, J=7.0 and 14.3 Hz, 2H), 7.17-7.26
Step 2:
A mixture of ethyl (3-bromophenyl)acetate (0.3 g, 1.24 mmol) and tetrabutylammonium fluoride hydrate (0.97 g, 3.72 mmol), was treated with PdCl2(PPh3)2 (26 mg, 0.037 mmol; 3 mole %) and 1-pentyne (367 372 mmol) in a sealed tube. The tube was heated at 80° C. for 2 h. The mixture was treated with water, and was extracted with diethyl ether. The organic extract was dried over sodium sulfate, filtered and evaporated in vacuo to give the crude product. Purification on a Biotage™ 25M column (silica), eluting with ethyl acetate/hexane 0:1 to 2:98, gave ethyl (3-(pentyne-1-yl)phenyl)acetate as a pale yellow oil (0.23 g, 79%).
Step 3:
To ethyl[3-[pentyne-1-yl]phenyl]-acetate (0.23 g, 0.98 mmol) in ethanol (5 mL) under nitrogen atmosphere was added Pd on carbon (10%, 25 mg, 10% w/w). The mixture was vigorously stirred under hydrogen atmosphere at room temperature overnight. The solution was filtered and the palladium/carbon was washed with ethanol (20 mL). The filtrate was concentrated with silica gel. The crude product was purified by flash chromatography using a mixture of 10% hexanes/ethyl acetate. A clear oil was obtained (0.21 g, 90%).
Step 4:
To a solution of the ester (0.2 g, 0.9 mmol) in tetrahydrofuran (5 mL), methanol (1.5 mL) and water (1.5 mL) was added lithium hydroxide (0.09 g, 3.6 mmol) at 0° C. The reaction mixture was stirred overnight at room temperature. Insolubles were filtered and the filtrate was concentrated under reduced pressure. The residue was than treated with 2M HCl and extracted with ethyl acetate. The organic phase was dried over sodium sulfate and evaporated under reduced pressure. The crude material was purified on a 40 L Biotage column (silica) using ethyl acetate/hexanes (0:10 to 4:6) as eluant. This gave pure (3-pentylphenyl)acetic acid (0.19 g, 99%) as a white gummy solid. 1H NMR (400 MHz, CD3OD): δ 0.90 (t, J=7.0 Hz, 3H), 1.28-1.38 (m, 4H), 1.61 (qt, J=7.6 Hz, 15.0 Hz, 2H), 2.58 (t, J=7.6 Hz, 2H), 3.56 (s, 2H), 7.07 (m, 3H), 7.20 (m, 1H); LRMS (ESI): m/z 207 (MH+); HPLC: 4.3 min.
Step 5:
To a stirred solution of the acid (0.19 g, 0.82 mmol) in ethanol (4 mL) and water (1 mL) was added sodium bicarbonate (0.07 g, 0.82 mmol). The reaction mixture was stirred at room temperature overnight. The solvent was evaporated and the white gummy solid was dissolved in water and the solution was lyophilized. This gave pure sodium salt of (3-pentylphenyl)acetic acid (0.17 g, 92%) as a white solid. mp 110-112° C.; 1H NMR (400 MHz, CD3OD): δ0.89 (t, J=6.8 Hz, 3H), 1.28-1.37 (m, 4H), 1.60 (qt, J=7.4 Hz, 15.0 Hz, 2H), 2.56 (t, J=7.6 Hz, 2H), 3.43 (s, 2H), 6.96 (m, 1H), 7.12 (m, 3H); LRMS (ESI): m/z 207 ((MH+); HPLC: 4.3 min.
The above compound was prepared as for compound I starting with E-(3-pent-1-enyl-phenyl)acetic acid methyl ester. The latter was prepared by reacting 3-bromophenyl acetic acid methyl ester with trans-1-pentenylboronic acid pinacol ester under Suzuki conditions. White solid; 1H NMR (400 MHz, CD3OD): δ=7.32 (s, 1H), 7.11-7.18 (m, 3H), 6.35 (d, J=15.7 Hz, 1H), 6.20-6.27 (m, 1H), 3.44 (s, 2H), 2.19 (m, 2H), 1.45-1.54 (m, 2H), 0.96 (t, J=7.4, 3H); 13C NMR (101 MHz, CD3OD): δ=179.26, 138.25, 137.92, 130.32, 130.04, 128.06, 127.59, 126.60, 123.52, 45.21, 35.06, 22.52, 12.89; LRMS (ESI): m/z 205 (MH+); HPLC: 4.1 min.
The above compound was prepared as for compound I starting with 5-bromo-2-methoxyphenylacetic acid methyl ester. Demethylation of the methoxy group was undertaken using a solution of boron tribromide (1M/CH2Cl2) at −78° C. for 1 h then at 0° C. during 20 min. White solid; 1H NMR (400 MHz, CD3OD): δ=6.88 (m, 2H), 6.71 (d, J=8.6 Hz, 1H), 3.50 (s, 2H), 2.49 (t, J=7.6 Hz, 2H), 1.54-1.62 (m, 2H), 1.29-1.38 (m, 4H), 0.91 (t, J=7.0 Hz, 3H); 13C NMR (101 MHz, CD3OD): δ=180.08, 154.04, 134.03, 130.26, 127.36, 124.15, 116.57, 42.48, 34.91, 31.60, 31.42, 22.45, 13.24; LRMS (ESI): m/z 177 (MH+—CO—NaOH); HPLC, 3.7 min.
The above compound was prepared as for compound I starting with E-methyl 3-(3-bromo-4-fluorophenyl)acrylate. The latter was prepared by mixing a solution of 3-bromo-4-fluorobenzaldehyde and ethoxycarbonylmethylenetriphenylphosphorane in dry dichloromethane at room temperature. White solid; 1H NMR (400 MHz, CD3OD): δ=6.67-6.74 (m, 2H), 6.58 (m, 1H), 2.49 (t, J=7.6 Hz, 2H), 2.23 (t, J=7.4 Hz, 2H), 2.15 (m, 2H), 1.25 (m, 2H), 0.99-1.06 (m, 4H), 0.61 (t, J=6.7 Hz, 3H); 13C NMR (101 MHz, D2O); δ=182.38, 160.69, 158.28, 137.37, 130.34, 129.58, 126.84, 114.99, 39.68, 31.51, 29.92, 28.90, 22.31, 16.66; LRMS (ESI): m/z 221 (MH+—H2O); HPLC: 4.5 min.
The above compound was prepared as for compound I starting with 3-Oxo-3-bromophenylpropionic acid ethyl ester. The ketone group and the double bond were simultaneously reduced using palladium/carbon in ethanol under hydrogen pressure. White solid; 1H NMR (400 MHz, CDCl3): δ 7.14-710 (m, 1H), 7.04-7.00 (m, 2H), 6.95-6.93 (m, 1H), 2.88-2.84 (m, 2H), 2.55 (t, J=7.4 Hz, 2H), 2.44-2.40 (m, 2H), 1.63-1.55 (m, 2H), 1.35-1.28 (m, 4H), 0.90 (m, 3H); 13C NMR (101 MHz, CD3OD): δ 179.3, 141.2, 140.8, 126.7, 126.4, 124.0, 123.8, 38.6, 34.2, 31.2, 29.9, 29.8, 20.9, 11.7; LRMS (ESI): m/z 203 (MH+—CO—NaOH); HPLC: 4.5 min.
Step 1:
A suspension of sodium hydride (60% w/w in mineral oil; 0.5 g, 13.6 mmol) in anhydrous THF (8 mL) was cooled to 0° C., and was treated with a solution of methyl[3-pentylphenyl]acetate (1.0 g, 4.5 mmol) in anhydrous THF (4 mL). The reaction was stirred at 0° C. for 60 min, and was then treated with methyl iodide (0.7 mL, 11.3 mmol). The reaction was allowed to warm slowly to room temperature, and was stirred at this temperature overnight. The reaction was quenched by addition of saturated aqueous ammonium chloride (10 mL), and the mixture was extracted with ether (3×20 mL). Combined extracts were dried over magnesium sulfate and evaporated to dryness. Purification on a silica pad, eluting with ethyl acetate/hexane 1:99 then 2:98, gave methyl 2-methyl-2-(3-pentylphenyl)propionate as a colorless oil (0.68 g, 60%). 1H NMR (400 MHz, CD3OD): δ 7.18-7.22 (m, 1H), 7.08-7.13 (m, 2H), 7.02-7.05 (m, 1H), 3.62 (s, 3H), 2.58 (t, J=7.6 Hz, 2H), 1.55-1.62 (m, 2H), 1.53 (s, 6H), 1.28-1.36 (m, 4H), 0.90 (t, J=7.1 Hz, 3H); HPLC: 5.5 min.
Step 2:
A solution of the ester in THF (8 mL), methanol (2 mL) and water (2 mL) was treated with lithium hydroxide (0.2 g, 8.2 mmol), and the reaction was stirred at room temperature overnight, than at 50° C. for 2 days, and at room temperature for 10 days. The reaction was filtered and the funnel was washed with methanol (2×20 mL). Combined filtrate and washings were treated with 2M HCl (7 mL), and the mixture was extracted with ethyl acetate (3×40 mL). Combined extracts were washed with water (2×30 mL), dried over sodium sulfate, filtered and evaporated in vacuo, to give 2-methyl-2-(3-pentylphenyl)propionic acid as a pale yellow syrup (0.64 g, 99%). This material was used without further purification. 1H NMR (400 MHz, CDCl3): δ 7.19-7.27 (m, 3H), 7.07-7.10 (m, 1H), 2.60 (t, J=7.8 Hz, 2H), 1.60 (s, 6H), 1.58-1.63 (m, 2H), 1.30-1.37 (m, 4H), 0.89 (t, J=7.0 Hz, 3H); LRMS (ESI): m/z 257 (MNa+); HPLC: 4.7 min.
Step 3:
A solution of the acid in ethanol (16 mL) was treated with water (4 mL) and sodium bicarbonate (0.2 g, 2.7 mmol), and the reaction was stirred at room temperature for 3 days. Solvent was evaporated in vacuo, and the residue was dissolved in water, filtered and lyophilized to give sodium 2-methyl-2-[3-pentylphenyl]propionate as a white solid (0.7 g, 96%). 1H NMR (400 MHz, CD3OD): δ 7.19-7.23 (m, 2H), 7.13 (dd, J=7.6, 7.6 Hz, 1H), 6.91-6.95 (m, 1H), 2.56 (t, J=7.7 Hz, 2H), 1.56-1.63 (m, 2H), 1.46 (s, 6H), 1.28-1.39 (m, 4H), 0.90 (t, J=7.0 Hz, 3H); 13C NMR (101 MHz, CD3OD): δ 184.35, 148.62, 142.13, 127.51, 126.14, 125.32, 123.16, 36.01, 31.57, 31.40, 27.45, 22.44, 13.22; LRMS (ESI): m/z 235; (M-Na++2H+); HPLC: 4.6 min.
The above compound was prepared as for compound VI except sodium hydride was replaced by diisopropylamine/n-butyllithium and methyl iodide by hydroxymethyl-1H-benzotriazole. White solid; 1H NMR (400 MHz, CDCl3): δ 7.19-7.14 (m, 3H), 7.01-6.98 (m, 1H), 4.01-3.96 (m, 1H), 3.72-3.57 (m, 1H), 3.31-3.30 (m, 1H), 2.58-2.55 (m, 2H), 1.64-1.56 (m, 2H), 1.37-1.29 (m, 4H), 0.90 (t, 3H, J=7.0 Hz); 13C NMR (101 MHz, CD3OD): δ 179.7, 142.7, 139.8, 128.4, 127.9, 126.3, 125.6, 65.2, 57.5, 35.8, 31.5, 31.3, 22.4, 13.2; LRMS (ESI): m/z 473 (2M-2Na++3H+); HPLC: 3.5 min.
The above compound was prepared as for compound I starting with 2-methyl-2-(3-pentylphenyl)malonic acid diethyl ester. The latter was prepared by reacting 2-(3-bromophenyl)malonic acid diethyl ester with methyl iodide followed by Suzuki coupling using trans-1-pentenyl-1-boronic acid pinacol ester then reduction of the double bond by hydrogenation. White solid; 1H NMR (400 MHz, CD3OD): δ 7.19-6.95 (m, 4H), 3.54 (q, J=7.0 Hz, 1H), 2.56 (t, J=7.6 Hz, 2H), 1.64-1.56 (m, 2H), 1.38 (d, J=7.2 Hz, 3H), 1.37-1.20 (m, 4H), 0.90 (t, J=7.0 Hz, 3H); 13C NMR (CD3OD): δ 182.2, 144.4, 142.5, 127.8, 127.6, 125.8, 124.7, 49.2, 35.9, 31.5, 31.3, 22.4, 19.0, 13.2; LRMS (ESI): m/z 221(M−Na++2H+); HPLC: 4.5 min.
The above compound was prepared as for Compound IV starting with E-methyl 3-(3-but-1-enylphenyl)acrylate. The latter was prepared by reacting isophthaldehyde with carbomethoxymethylenetriphenylphosphorane followed by a Wittig reaction using n-butyltriphenylphosphonium bromide. White solid; 1H NMR (400 MHz, CDCl3): δ 7.10 (t, 1H, J=7.5 Hz), 7.01 (s, 1H), 6.94 (d, 2H, J=7.0 Hz), 2.68 (t, 2H, J=7.9 Hz), 2.43 (t, 2H, J=7.7 Hz), 2.29 (t, 2H, J=7.9 Hz), 1.40 (m, 2H, J=7.4 Hz), 1.14 (m, 2H, J=7.4 Hz), 0.72 (t, 3H, J=7.4 Hz); 13C NMR (101 MHz, CD3OD): δ 142.7; 142.4; 128.2; 128.0; 125.6; 125.4; 125.3; 40.1; 35.5; 33.9; 32.7; 22.2; 13.1; LRMS (ES1): m/z 209 (MH+); HPLC: 4.1 min.
Step 1:
To a solution of (3-bromophenyl)acetic acid (12.2 g, 56.8 mmol) in methanol (150 mL) was added p-toluenesulfonic acid (5.4 g, 28.4 mmol). The reaction mixture was stirred at reflux for 3 hours. The solvent was evaporated and the residue was dissolved in a mixture of ethyl acetate/water (3:2). The organic layer was dried over sodium sulfate and concentrated. The residue was purified using a silica pad eluting with a mixture of hexanes/ethyl acetate (9:1). This gave (3-bromophenyl)acetic acid methyl ester as a colorless oil (11.7 g, 90%). 1H NMR (400 MHz, CD3OD): δ=7.46 (m, 1H), 7.41 (m, 1H), 7.22 (m, 2H), 3.68 (s, 3H), 3.65 (s, 2H); LRMS (ESI): m/z=229 (MH+); HPLC: 3.8 min.
Step 2:
To a solution of the ester (6.0 g, 26.2 mmol) in tert-butanol (24 mL) was added, under nitrogen atmosphere, sodium iodide (7.8 g, 52.4 mmol), N,N′-dimethylethylenediamine (0.3 mL, 2.6 mmol) and copper iodide (0.3 g, 1.3 mmol). The reaction mixture was heated in a microwave apparatus at 145° C. for 1 h. Water (100 mL) was added and the product was extracted with ethyl acetate (3×50 mL). The organic layer was dried over sodium sulfate and concentrated. The residue was purified by flash chromatography on silica gel with a mixture of hexanes/ethyl acetate (8:2). This gave 3-iodophenylacetic acid methyl ester as a colorless oil (6.6 g, 86%). 1H NMR (400 MHz, CDCl3): δ=7.63 (m, 1H), 7.58-7.61 (m, 1H), 7.23-7.26 (m, 1H), 7.05 (dd, J=7.8 Hz, 1H), 3.69 (s, 3H), 3.56 (s, 2H); LRMS (ESI): m/z=277 (MH+).
Step 3:
The iodoester (6.2 g, 22.5 mmol) was mixed with palladium chloride (0.16 g, 0.22 mmol), triphenylphosphine (59.0 mg, 0.22 mmol) and diethylamine (60 mL) under nitrogen atmosphere. To this mixture was added copper(I) iodide (43 mg, 0.22 mmol) and propargyl alcohol (1.57 g, 28.1 mmol) and the reaction mixture was stirred overnight at 45° C. Diethylamine was removed under reduced pressure and 100 mL of water was added. The mixture was then extracted with ethyl acetate (3×30 mL) and the crude product was purified by flash chromatography using a mixture of ethyl acetate/hexanes (30%). This gave pure [3-(3-hydroxyprop-1-ynyl)phenyl]acetic acid methyl ester as a brownish oil (3.8 g, 84%). 1H NMR (400 MHz, CDCl3): δ=7.33-7.37 (m, 2H), 7.23-7.30 (m, 2H), 4.49 (d, J=6.1 Hz, 2H), 3.69 (s, 3H), 3.60 (s, 2H), 1.68 (t, J=6.3 Hz, 1H); LRMS (ESI): m/z=227 (MNa+); HPLC: 2.7 min.
Step 4:
To the methyl ester (3.8 g, 18.7 mmol) in ethanol (70 mL) under nitrogen atmosphere was added 10% palladium/carbon (0.30 g). The atmosphere was changed for hydrogen. The mixture was vigorously stirred at room temperature overnight. The solution was filtered and the palladium/carbon was washed with ethanol (50 mL). The filtrate was concentrated and the crude product was purified by flash chromatography using a mixture of hexanes/ethyl acetate (3:2). This gave pure 3-(3-hydroxypropyl)phenyl]acetic acid methyl ester as a colorless oil (3.20 g, 82%). 1H NMR (400 MHz, CD3OD): δ=7.21 (t, J=7.6 Hz, 1H), 7.11 (s, 1H), 7.07 (m, 2H), 3.67 (s, 3H), 3.61 (s, 2H), 3.56 (t, J=7.6 Hz, 2H), 2.66 (t, J=7.6 Hz, 2H), 1.78-185 (m, 2H); LRMS (ESI): m/z=209 (MH+); HPLC: 2.6 min.
Step 5:
At 0° C., under nitrogen atmosphere, pyridinium chlorochromate (1.44 g, 6.70 mmol) and molecular sieves were added to a solution of the methyl ester (0.9 g, 4.4 mmol) in dry dichloromethane (20 mL). The reaction mixture was stirred for 20 min at 0° C. and 3 h at room temperature. Ether (20 mL) was added and the precipitate was filtered and washed with ether (40 mL). The filtrate was evaporated to give [3-(3-oxopropyl)phenyl]acetic acid methyl ester as a brownish oil (0.9 g, 97%). The aldehyde was used in the next step without further purification. 1H NMR (400 MHz, CDCl3): δ=9.82 (t, J=1.4 Hz, 1H), 7.24-7.28 (m, 2H), 7.11 (m, 2H), 3.69 (s, 3H), 3.60 (s, 2H), 2.95 (t, J=7.6 Hz, 2H), 2.80 (t, J=7.0 Hz, 2H).
Step 6:
The aldehyde (0.9 g, 4.3 mmol) was dissolved in tetrahydrofuran (9 mL). In a separate flask containing a solution of (ethyl)triphenylphosphonium bromide (2.1 g, 5.6 mmol) in dry tetrahydrofuran (17 mL) at −10° C. was added a solution of 2.3 M n-butyllithium (1.94 mL, 5.8 mmol). The orange solution was stirred at this temperature for 20 min and at 0° C. for 40 min. To this solution was added the aldehyde and the mixture was stirred for 1 h at 0° C. and at room temperature overnight. Water (30 mL) was then added and the organic layer was extracted with ether (3×30 mL). The combined ether layers were washed with brine and dried. The solvent was evaporated and the residue was purified using a mixture of petroleum ether/ethyl acetate (95%) as eluent. This gave pure E/Z-(3-pent-3-enyl-phenyl)acetic acid methyl ester as a colorless oil (0.25 g, 27%). 1H NMR (400 MHz, CDCl3): d=7.13-7.18 (m, 1H), 7.06-7.08 (m, 3H), 5.31-5.44 (m, 2H), 3.62 (s, 3H), 3.52 (d, J=7.2 Hz, 2H), 2.57 (t, J=7.8 Hz, 2H), 2.25-2.31 (m, 2H), 1.57 (dd, J=3.3, 1.4 Hz, 3H).
Step 7:
To a solution of the olefin (0.13 g, 0.60 mmol) in tetrahydrofuran (3 mL), methanol (1.5 mL) and water (1.5 mL) was added lithium hydroxide (73 mg, 3.1 mmol) at 0° C. The reaction mixture was stirred overnight at room temperature. The solvent was concentrated, acidified with 2M HCl and extracted with ethyl acetate (3×15 mL). The organic phase was dried and evaporated under high vacuum. The crude product was purified on a silica pad with ethyl acetate/hexanes (20%). This gave pure E/Z-(3-Pent-3-enylphenyl)acetic acid (0.12 g, 100%) as colorless oil. 1H NMR (400 MHz, CDCl3): δ=10.70-11.50 (br s, 1H), 7.26-7.30 (m, 1H), 7.13-7.20 (m, 3H), 5.44-5.53 (m, 2H), 3.65 (s, 2H), 2.67-2.71 (m, 2H), 2.33-2.42 (m, 2H), 1.58-1.68 (m, 3H).
Step 8:
To a stirred solution of the acid (0.12 g, 0.6 mmol) in ethanol (3 mL) and water (2 mL) was added sodium bicarbonate (50 mg, 0.6 mmol). The reaction mixture was stirred at room temperature overnight. The solvent was concentrated and the residue was diluted in water (70 mL) and the solution was lyophilized. This gave pure sodium salt E/Z-(3-pent-3-enylphenyl)acetic acid as a white solid (0.14 g, 90%). 1HNMR (400 MHz, D2O): (major, E-isomer) δ=7.12 (dd, J=7.4 Hz, 1H), 7.00 (s, 1H), 6.99 (d, J=7.4 Hz, 1H), 6.95 (d, J=7.6 Hz, 1H), 5.27-5.38 (m, 2H), 3.33 (s, 2H), 2.53-2.48 (m, 2H), 2.13-2.24 (m, 2H), 1.35-1.44 (m, 3H).
Step 1:
A solution of methyl[3,5-dihydroxyphenyl]acetate (2.1 g, 11.5 mmol) in acetone (100 mL) was treated with potassium carbonate (2.4 g, 17.4 mmol), potassium iodide (0.38 g, 2.31 mmol) and benzyl bromide (1.5 mL, 12.7 mmol), and the mixture was stirred at room temperature overnight. The reaction was diluted with water, and was extracted with dichloromethane (×3). Combined organic extracts were dried over sodium sulfate and evaporated in vacuo. The crude material was purified on a Biotage™ 40M column (silica), eluting with 40% ethyl acetate/hexane, to give methyl[3-benzyloxy-5-hydroxyphenyl]acetate (1.0 g, 33%). 1H NMR (400 MHz, CDCl3): δ 7.32-7.42 (m, 5H), 6.48
Step 2:
A solution of the benzyl ether (1.04 g, 3.8 mmol) in CH2Cl2 (15 mL) at 0° C., was treated with N-phenyl-bis(trifluorosulfonyl)imide (1.40 g, 3.9 mmol), and then triethylamine (0.6 mL, 4.1 mmol) was added slowly. The reaction was stirred at 0° C. for 1 h, and than at room temperature for 1 h. The reaction mixture was diluted with water, and than extracted with diethylether (×2). Combined organic extracts were washed with 1M aqueous sodium hydroxide, water (×2) and saturated aqueous sodium chloride, then dried over sodium sulfate, filtered and evaporated in vacuo, to give the crude product. Purification on a Biotage™ 40M column (silica), eluting with ethyl acetate/hexane 0:1 to 1:4, gave methyl[3-benzyloxy-5-trifluoromethanesulfonyloxyphenyl]acetate (1.2 g, 79%). 1H NMR (400 MHz, CDCl3): δ 7.36-7.46 (m, 5H), 6.98 (s, 1H), 6.97 (s, 1H), 6.84 (s, 1H), 5.06 (s, 2H), 3.72 (s, 3H), 3.63 (s, 2H).
Step 3:
A solution of E-1-penten-1-ylboronic acid pinacol ester (0.8 g, 3.9 mmol) in dimethoxyethane (5 mL) was treated with a solution of the triflate (1.2 g, 3.0 mmol) in dimethoxyethane (5 mL). The solution was treated with palladium zero (0.7 g, 0.6 mmol) and 2M aqueous sodium carbonate (1.3 mL, 2.6 mmol). The mixture was then heated at 90° C. for 3 days. The reaction was cooled to room temperature and filtered through celite. The filtrate was evaporated in vacuo, and the crude material was purified on a Biotage™ 25M column (silica), eluting with ethyl acetate/hexane 0:1 to 5:95, to give methyl[3-benzyloxy-5-[pent-1-enyl]phenyl]acetate (0.4 g, 40%). 1H NMR (400 MHz, CDCl3): δ 7.36-7.47 (m, 5H), 6.90-6.92 (m, 2H), 6.79 (dd, J=2.0, 2.0 Hz, 1H), 6.35 (d, J=15.9 Hz, 1H), 6.24 (dt, J=15.9, 6.8 Hz, 1H), 5.07 (s, 2H), 3.70 (s, 3H), 3.59 (s, 2H), 2.20 (td, J=7.4, 6.8 Hz, 2H), 1.51 (dt, J=7.4 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H).
Step 4:
A solution of the alkene (0.4 g, 1.2 mmol) in ethanol (13 mL) was treated with 1% palladium on carbon (40 mg). The mixture was stirred under 1 atm. of hydrogen at room temperature overnight. The reaction was filtered, evaporated in vacuo, and purified on a Biotage™ 25S column (silica), eluting with ethyl acetate/hexane 0:1 to 15:85 to give methyl[3-hydroxy-5-pentylphenyl]acetate (0.3 g, 93%). 1H NMR (400 MHz, CDCl3): δ 6.64 (s, 1H), 6.58-6.60 (m, 2H), 3.70 (s, 3H), 3.55 (s, 2H), 2.51 (t, J=7.7 Hz, 2H), 1.55-1.59 (m, 2H), 1.28-1.34 (m, 4H), 0.88 (t, J=7.0 Hz, 3H).
Step 5:
A solution of the ester (0.3 g, 1.3 mmol) in ethanol (12 mL) was treated with water (3 mL) and lithium hydroxide (155 mg, 6.4 mmol), and the mixture was stirred vigorously at room temperature overnight. The reaction mixture was diluted with water (100 mL); washed with dichloromethane; then acidified to pH 1 with 1M aqueous HCl and extracted with dichloromethane (×3). Combined organic extracts were dried over sodium sulfate (0.3 g, 95%). This material was used without further purification. 1H NMR (400 MHz, CDCl3): δ 6.66 (s, 1H), 6.58-6.59 (m, 2H), 3.55 (s, 2H), 2.52 (t, J=7.7 Hz, 2H), 1.55-1.59 (m, 2H)
Step 6:
A solution of the acid (0.27 g, 1.23 mmol) in ethanol (6 mL) and water (6 mL) was treated with a sodium bicarbonate (0.1 g, 1.2 mmol), and the reaction was stirred at room temperature for a few hours. Solvent was concentrated in vacuo, and the solution was diluted with water, filtered (0.2 μm), and lyophilized to give sodium [3-hydroxy-5-pentylphenyl]acetate as a white solid (0.3 g, 95%). mp 63-66° C.; 1H NMR (400 MHz, CD3OD): δ 6.63 (s, 1H), 6.58 (s, 1H), 6.42 (s, 1H), 3.36 (s, 2H), 2.48 (t, J=7.6 Hz, 2H), 1.55-1.62 (m, 2H), 1.26-1.38 (m, 4H), 0.89 (t, J=6.8 Hz, 3H); 13C NMR (101 MHz, CD3OD): δ 177.79, 155.31, 142.36, 137.62, 119.08, 111.66, 111.18, 43.70, 34.17, 29.95, 29.56, 20.87, 11.64; LRMS (ESI): m/z 445.2 (2M-2Na++3H+), m/z 223 (M−Na++2H+); HPLC: 3.5 min.
The above compound was prepared as for compound I starting with 4-pentylbenzoic acid. White solid; 1H NMR (400 MHz, D2O): δ 7.61 (d, J=8.3 Hz, 2H), 7.12 (d, J=8.5 Hz, 2H), 2.46 (t, J=7.5 Hz, 2H), 1.38-1.45 (m, 2H), 1.04-1.15 (m, 4H), 0.65 (t, J=7.0 Hz, 3H); 13C NMR (101 MHz, D2O): δ 175.79, 147.29, 133.55, 129.15, 128.47, 35.07, 30.81, 30.45, 22.00, 13.42; LRMS (ESI): m/z 193 (M−Na++2H+); HPLC: 43 min.
The above compound was prepared as for compound IX starting with 3-[hex-1-enyl]benzoate. The latter was prepared by reacting pentyltriphenylphosphonium bromide with methyl 3-formylbenzoate. White solid; 1H NMR (400 MHz, CD3OD): δ 7.74-7.79 (m, 2H), 7.20-7.36 (m, 2H), 2.63 (t, J=7.6 Hz, 2H), 1.61-1.65 (m, 2H), 1.28-1.36 (m, 6H), 0.89 (t, J=7.5 Hz, 3H); 13C NMR (101 MHz, CD3OD): δ 174.64, 142.29, 137.65, 130.28, 129.13, 127.47, 126.50, 35.73, 31.74, 31.55, 28.89, 22.52, 13.28; LRMS (ESI): /m/z 307 (M−Na++2H+); HPLC: 3.0 min.
Step 1:
To a solution of indan-2-carboxylic acid (2.50 g) in ethanol (77 mL) at 25° C. was added concentrated H2SO4 (9 mL). The colorless solution was stirred at 75° C. over a period of 48 h. Solvent was removed under reduced pressure and the residue was diluted with dichloromethane (10 mL) and water (10 mL). The pH of the solution was brought from 1 to 14. Two layers were separated and the aqueous phase was extracted with ethyl acetate (30 mL). Organic layers were combined, dried over magnesium sulfate, filtered and concentrated under high vacuum. The solid obtained was purified on a Biotage™ 40M column (silica, hexanes/ethyl acetate 1:0 to 97:3) to give indan-2-carboxylic acid ethyl ester. Off-white oil; 1H NMR (400 MHz, CDCl3): δ 7.22-7.14 (m, 4H), 4.19 (q, J=7.04 Hz, 4H), 3.36-3.10 (m, 5H), 1.29 (t, J=7.04 Hz, 3H); LRMS (ESI): m/z 190 (MH+); HPLC: 4.3 min.
Step 2:
Butyryl chloride (0.8 mL, 7.9 mmol) was added to a mixture of the ethyl ester and AlCl3 (2.5 g, 18.5 mmol) in dichloromethane (20 mL). After stirring at room temperature for 5 h, the reaction was poured into a mixture ice and 1N HCl. The aqueous phase was extracted with dichloromethane (30 mL). The combined organic extracts were dried over magnesium sulfate, filtered and evaporated under reduced pressure. The crude residue was purified on a Biotage™ 40 L column (silica, hexanes/ethyl acetate 1:0 to 8:2) to yield 5-butyrylindan-2-carboxylic acid ethyl ester as colorless oil (1.0 g, 50%); 1H NMR (400 MHz, CDCl3): δ 7.80-7.77 (m, 2H), 7.27 (d, J=8.61 Hz, 1H), 4.18 (q, J=7.04 Hz, 4H), 3.36-3.10 (m, 5H), 2.91 (t, J=7.24 Hz, 2H), 1.78-1.65 (m, 2H), 1.28 (t, J=7.04 Hz, 3H), 0.99 (t, J=7.24 Hz, 3H); LRMS (ESI): m/z 261 (MH+); HPLC: 4.5 min.
Step 3:
To a stirred solution of the ketone (0.56 g, 2.17 mmol) in 1,2-dichloroethane (11 mL) at room temperature was added zinc iodide (1.04 g, 3.25 mmol) and sodium cyanoborohydride (1.0 g, 16.3 mmol). The reaction mixture was stirred at 25° C. for a period of 20 h. The reaction was filtered through a celite pad and the solvent was evaporated under reduced pressure. The crude residue was purified on a Biotage™ 25M column (silica, hexanes/ethyl acetate 1:0 to 8:2) to yield 5-butylindan-2-carboxylic acid ethyl ester (0.4 g, 77%). 1H NMR (400 MHz, CDCl3): δ 7.11 (d, J=7.83 Hz, 1H), 7.04 (s, 1H), 6.98 (d, J=7.63 Hz, 1H), 4.18 (q, J=7.24 Hz, 2H), 3.37-3.14 (m, 5H), 2.57 (t, J=7.63 Hz, 2H), 1.62-1.54 (m, 2H), 1.40-1.27 (m, 5H), 0.93 (t, J=7.24 Hz, 3H); HPLC: 5.5 min.
Step 4:
To a solution of the ester (0.4 g, 1.65 mmol) in a mixture of acetonitrile/methanol/water (5 mL, 3:1:1) was added lithium hydroxide (0.3 g, 10.7 mmol). The reaction was stirred at 25° C. for 15 h. After evaporation of solvent, the residue was extracted with dichloromethane (30 mL). The aqueous phase was acidified with 1N HCl until pH=4, and was then extracted with dichloromethane (2×25 mL). The organic extracts were dried over magnesium sulfate, filtered and evaporated under reduced pressure. The crude residue was purified on a Biotage™ 12M column (silica, hexanes/ethyl acetate 1:0 to 7:3) to yield 5-butylindan-2-carboxylic acid (0.3 g, 90%). HPLC: 4.3 min. To a solution of the acid (0.32 g, 1.49 mmol), in a mixture of acetonitrile/water (5 mL, 2:3) was added sodium bicarbonate. The reaction was stirred overnight at room temperature. After evaporation of acetonitrile, the residue was diluted with water (4 mL). The solution was filtered through a 0.45 μM filter and lyophilized. This gave pure sodium salt of 5-butylindan-2-carboxylic acid. White solid; 1H NMR (400 MHz, CD3OD): δ 7.02 (d, J=7.63 Hz, 1H), 6.96 (s, 1H), 6.88 (d, J=7.63 Hz, 1H), 3.18-3.04 (m, 5H), 2.54 (t, J=7.63 Hz, 2H), 1.59-1.51 (m, 2H), 1.37-1.28 (m, 2H), 0.92 (t, J=7.24 Hz, 3H); 13C NMR (101 MHz CD3OD): δ 183.2, 143.1, 140.7, 140.2, 126.2, 123.9, 123.6, 37.2, 36.9, 35.4, 34.1, 22.1, 13.1; LRMS (ESI): m/z 201 (MH+—NaOH); HPLC: 4.3 min.
Female C57BL/6 mice, 6 to 8 week old, were immunosuppressed by treatment with 200 mg/kg of cyclophosphamide administered intravenously at day 0. To examine the immunoprotective effect of compound I, mice were pre-treated orally at day −3, −2 and −1 with the compound. Mice were sacrificed at day +5 by cardiac puncture and cervical dislocation. Then, a gross pathological observation of the femurs (as a source of bone marrow cells) was recorded. After the sacrifice, tissues were crushed in PBS buffer and cells were counted on a hemacytometer.
A significant increase in total bone marrow cell count was observed with oral pre-treatment with compound I in cyclophosphamide treated mice (
Also, an increase in red bone marrow cell count was observed with oral pre-treatment with compound I in cyclophosphamide immunosuppressed mice (Table 1). Furthermore, compound I increases circulating red blood cells.
Additional compounds related to compound I were also prepared and tested for biological activity. Table 2 summarizes the activity of those additional compounds. The results in this table are expressed depending on their degree of activity on hematopoiesis/erythropoiesis compared to the activity of compound I where: ++++ designates greater activity than compound I, +++ designates 70 to 100% of compound I activity, ++ designates 40-70% of compound I activity, + designates 5-40% of compound I activity and “n/a” indicates that the compound has not been tested.
LPS-induced inflammation in the rat air-pouch model is believed to mimic the pathological process occurring in joint diseases such as arthritis. This is because the connective tissues formed along the air pouch are similar to those found in chronic joint diseases. LPS-induced inflammation and chronic joint diseases share other features, including markedly elevated PGE2, neutrophil infiltration, cytokine formation, and tissue damage.
An air cavity was produced at day −6 by subcutaneous injection of 20 ml of sterile air into the intracapsular area of the back of male Lewis rats (175-200 g). An additional 10 ml of air was injected into the cavity at day −3 to keep the space open. At day 0, compounds were administered intravenously and one hour later lipopolysaccharide (LPS: 2.5 ml, 2 μg/ml in PBS) was injected into the pouch to produce an inflammatory reaction. Two hours after treatment with LPS, animals were euthanized by CO2 asphyxiation and 5 ml of PBS/heparin (10 U/ml)/indomethacin (36 μg/ml) was injected into the pouch. The pouch fluid was collected and PGE2 was determined in the pouch exudates by ELISA.
As illustrated in
Oxidative stress and inflammation are related to several chronic diseases including but not limited to cardiovascular diseases, cancer, diabetes, arthritis. Alzheimer's disease and auto-immune disease. Nitric oxide (NO), produced by nitric oxide synthase, has been identified as an important molecule involved in inflammation and sepsis. Inducible nitiric oxide synthase (iNOS) is not expressed under normal conditions. However, after exposure to endogenous and exogenous stimulators, it can be induced in various cells such as macrophages, smooth muscle cells and hepatocytes, to trigger several disadvantageous cellular responses, such as inflammation. Hence, the level of iNOS may reflect the degree of inflammation, thereby permitting an evaluation of the effects of drugs on the inflammatory process.
The effect of selected compounds on NO production was undertaken in RAW264.7 (macrophage-like) cells. RAW264.7 cells were cultured with 1 μg/ml of LPO and 0.5 ng/ml of interferon in presence or absence of compounds for 16 hours in a humidified atmosphere of 95% air-5% carbon dioxide at 37° C. Nitric oxide measurement in the culture medium was measured using the Griess reagent after an incubation of 30 minutes at room temperature. The absorbance at 548 nm was read and compared with standard solutions of NaNO2. Cell viability was assessed with the addition of 50 μl of MTT. After an incubation of 4 hours, the medium was removed and 150 μl of DMSO was added to dissolve the crystals. The optical density of each sample was read at 570 nm against a blank prepared from cell-free wells.
Demonstration of the in vivo protection effect of Compound I on renal tissue was undertaken in the 5/6 nephrectomized (Nx) rat model using the following procedure. Male 6 week-old Wistar rats were subjected to 5/6 nephrectomy or sham operations. Under fluothane anesthesia, renal ablation was achieved by removing two-thirds of the left kidney followed by a right unilateral nephrectomy 7 days later. Sham rats underwent exposition of the kidneys and removal of the perirenal fat. Twenty-one days after the first operation, rats were randomized in the study by their reduced glomerular filtration rate (GFR) of creatinine indicating a dysfunction of the kidney. Animals that underwent the sham operation were given vehicle (saline) and were used as controls. Nx animals were divided in groups receiving the vehicle or Compound I. Saline or Compound I was given by gastric gavage once daily up to the sacrifice. GFR was measured every three weeks in order to assess the severity of this end-stage renal disease model. Rats were sacrificed at day 190.
Demonstration of the in vivo heart protection effect of Compound I was undertaken in the 5/6 nephrectomized (Nx) rat model using the procedure described in example 1. Briefly, heart pressure was recorded with a RTBP2000™ apparatus (Kent Scientific) in 5/6 nephrectomized rats to demonstrate that Compound I exerts a protective effect on the heart in severely affected 5/6 nephrectomized rats. A significant decrease in blood pressure was observed in Compound I-treated Nx rats (
Demonstration of the in vivo protection effect of oral administration of Compound I was undertaken in the doxorubicin-induced nephrotoxicity model using the following procedure C57BI/6 mice (6-10 week-old) were treated with Compound I prophylacticly from day −3 to day 10 or treated therapeutically from day 1 to 10. Nephrotoxicity was induced by an intravenous injection of 10 mg/kg of doxorubicin at day 0, Serum albumin and creatinine were monitored at days 4, 7, 9 and 11.
Prophylactic treatment with Compound I inhibits the decrease of serum albumin induced by doxorubicin. Therapeutic treatment with Compound I has no effect on serum albumin level induced by doxorubicin. (
Prophylactic treatment with Compound I inhibits the increase of serum creatinine induced by doxorubicin. Therapeutic treatment with Compound I also inhibits the increase of serum creatinine level induced by doxorubicin. (
Doxorubicin is well known to induce nephro- and cardiotoxicity.
Doxorubicin induces early lesions primarily at the tubular region. Toxicity is further extended to the glomerulus (around day 11 post-doxorubicin).
The mechanism by which Compound I appears to protect against doxorubicin-induced nephrotoxicity involves inhibition of fibrosis as demonstrated by the significant inhibition of CTGF expression in kidneys treated with Compound I.
CTGF is also regulated by TGF-β.
This application is a continuation-in-part application of International Application No. PCT/CA2010/000677, filed May 3, 2010; which claims the benefit of U.S. provisional application Ser. No. 61/175,235, filed May 4, 2009, both of which are incorporated herein by reference in their entirety. This application also claims priority to International Application No. PCT/CA2011/001179, entitled Compounds and Compositions for the Treatment of Cancer, filed Oct. 26, 2011; which claims the benefit of U.S. provisional application Ser. No. 61/407,069, filed Oct. 27, 2010, both of which are incorporated herein by reference in their entirety.
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| Number | Date | Country | |
|---|---|---|---|
| 20120088784 A1 | Apr 2012 | US |
| Number | Date | Country | |
|---|---|---|---|
| 61175235 | May 2009 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/CA2010/000677 | May 2010 | US |
| Child | 13289592 | US |
