(19)
(11)EP 3 152 188 B1

(12)EUROPEAN PATENT SPECIFICATION

(45)Mention of the grant of the patent:
13.02.2019 Bulletin 2019/07

(21)Application number: 15744724.4

(22)Date of filing:  05.06.2015
(51)International Patent Classification (IPC): 
C07C 233/63(2006.01)
A61K 31/165(2006.01)
C07C 235/40(2006.01)
(86)International application number:
PCT/US2015/034566
(87)International publication number:
WO 2015/188152 (10.12.2015 Gazette  2015/49)

(54)

NOVEL EP4 AGONISTS AS THERAPEUTIC COMPOUNDS

NEUARTIGE EP4-AGONISTEN ALS THERAPEUTISCHE VERBINDUNGEN

NOUVEAUX AGONISTES DES RÉCEPTEURS EP4 UTILISÉS COMME COMPOSÉS THÉRAPEUTIQUES


(84)Designated Contracting States:
AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

(30)Priority: 06.06.2014 US 201462009028 P

(43)Date of publication of application:
12.04.2017 Bulletin 2017/15

(73)Proprietor: ALLERGAN, INC.
Irvine, CA 92612 (US)

(72)Inventors:
  • OLD, David W.
    Irvine, California 92620 (US)
  • HEIN, Christopher D.
    Irvine, California 92612 (US)

(74)Representative: Hoffmann Eitle 
Patent- und Rechtsanwälte PartmbB Arabellastraße 30
81925 München
81925 München (DE)


(56)References cited: : 
WO-A1-2006/004191
US-A- 4 540 690
US-B1- 6 410 781
WO-A1-2011/053870
US-A- 5 420 343
  
  • DATABASE REGISTRY [Online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 24 March 2014 (2014-03-24), XP002744512, Database accession no. 1572391-58-2
  • DATABASE REGISTRY [Online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 28 March 2014 (2014-03-28), XP002744513, Database accession no. 1575419-94-1
  
Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention).


Description

FIELD



[0001] The present invention relates to the compound and the compound for use in methods for treating disorders and specifically to agonists and antagonists for EP4 receptors as defined in the claims.

BACKGROUND



[0002] The prostanoid EP4 receptor is a G protein-coupled receptor that mediates the actions of prostaglandin E2 (PGE2) and is characterized by the longest intracellular C terminus loop when compared to other prostanoid receptors. The EP4 receptor is one of four receptor subtypes of prostaglandin E2 receptors. In general, EP4 receptors couple to G proteins and mediate elevations in cyclic-adenosine monophsophate ("cAMP") concentration, although they do participate in other pathways as well. Expression of EP4 receptors is controlled by various physiological and pathophysiological processes as these receptors participate in ovulation and fertilization, induce bone formation, T cell factor signaling, protect against inflammatory bowel disease, facilitate Langerhans cell migration and maturation and mediate joint inflammation in a model of collagen-induced arthritis, among others.

[0003] U.S. Pat. No. 6,552,067 teaches the use of prostaglandin EP4 selective agonists for the treatment of methods of treating conditions which present with low bone mass, particularly osteoporosis, frailty, an osteoporotic fracture, a bone defect, childhood idiopathic bone loss, alveolar bone loss, mandibular bone loss, bone fracture, osteotomy, bone loss associated with periodontitis, or prosthetic ingrowth in a mammal.

[0004] U.S. Pat. No. 6,586,468 teaches that prostaglandin EP4 agonists may be useful for the prophylaxis and/or treatment of autoimmune disorders such as amyotrophic lateral sclerosis, multiple sclerosis, Sjoegren's syndrome, arthritis, rheumatoid arthritis, systemic lupus erythematosus, post-transplantation graft rejection, asthma, abnormal bone formation, neurocyte death, pulmopathy, hepatopathy, acute hepatitis, nephritis, renal insufficiency, hypertension, myocardial ischemia, systemic inflammatory syndrome, pain induced by ambustion, sepsis, hemophagocytosis syndrome, macrophage activation syndrome, Still's diseases, Kawasaki diseases, bums, systemic granuloma, ulcerative colitis, Crohn's diseases, hypercytokinemia at dialysis and multiple organ failure and shock. WO2006004191 discloses certain heterocyclic amides for the treatment of several diseases such as glaucoma.

[0005] Inflammatory bowel disease constitutes a group of diseases characterized by inflammation of the large and small intestines and manifests symptoms such as diarrhea, pain, and weight loss. Kabashima and colleagues taught that "EP4 works to keep mucosal integrity, to suppress the innate immunity, and to down regulate the proliferation and activation of CD4+ T cells. These findings have not only elucidated the mechanisms of IBD by NSAIDs, but also indicated the therapeutic potential of EP4-selective agonists in prevention and treatment of IBD." (Kabashima, et. al., The Journal of Clinical Investigation, April 2002, Vol. 9, 883-893).

[0006] Various other diseases are mediated by the EP4 receptor such as esophageal ulcers, alcohol gastropathy, duodenal ulcers, non-steroidal anti-inflammatory drug-induced gastroenteropathy and intestinal ischemia. New compounds for use in methods for treating or preventing such diseases are desired.

SUMMARY



[0007] The present invention is directed in part to EP4 agonists and antagonists and their use in treating a variety of pathological conditions associated with activity of the EP4 receptors as defined in the claims.

[0008] The invention includes compounds having the following structure:

or a pharmaceutically acceptable salt or diastereomer or enantiomer thereof, wherein:

the dashed line is a single or a double bond;

Y is H or CH3;

X is zero (meaning completely absent), one or two substituents on the cycloalkane or cycloalkene ring, said substituents chosen from the group consisting of H, C1 - C6 alkyl, hydroxyalkyl, aryl, halogen, cycloalkene, OR, CF3, C(O)R, COCF3, SO2N(R)2, SO2NH2, NO2, and CN;

n is selected from the group consisting of 0, 1, 2, 3, or 4;

R is C1 - C6 alkyl;

R2 is selected from the group consisting of H, hydroxyalkyl, C1 - C6 alkyl, phenyl or biphenyl;

R3 is selected form the group consisting of H, C(O)R5, SO2R5, C1 - C6 alkyl;

R5 is C1-C6 alkyl, haloalkyl including trifluoromethyl, aryl or heteroaryl;

R6 is CO2H, CH3, CO2R2, CH2OR2, CONR2R3 or tetrazol-5-yl;

R7 is selected from the group consisting of H, CF3, OCH3; and

R8 is selected form the group consisting of H and OCH3.



[0009] Another embodiment of the invention includes compounds having the following structure:

or a pharmaceutically acceptable salt or diastereomer or enantiomer thereof, wherein:

the dashed line is a single or a double bond;

Y is H or CH3;

n is selected from the group consisting of 0, 1, 2, 3, or 4;

R2 is selected from the group consisting of H, C1 - C6 alkyl, hydroxyalkyl, phenyl or biphenyl;

R3 is selected form the group consisting of H, C(O)R5, SO2R5, C1 - C6 alkyl;

R5 is C1-C6 alkyl, haloalkyl including trifluoromethyl, aryl or heteroaryl;

R6 is CO2H, CH3, CO2R2, CH2OR2, CONR2R3 or tetrazol-5-yl;

R7 is selected from the group consisting of H, CF3, OCH3; and

R8 is selected form the group consisting of H and OCH3.



[0010] Some of the compounds of the present invention are diastereomers yet have very different activities, see Examples 9 and 10 and 12 and 13 in Table 1.

DETAILED DESCRIPTION



[0011] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention claimed. As used herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, "or" means "and/or" unless stated otherwise. Furthermore, use of the term "including" as well as other forms, such as "includes," and "included," is not limiting.

[0012] As used herein "alkane" refers to compounds consisting of only hydrogen and carbon, fully saturated and containing only single bonds.

[0013] As used herein, "alkyl" refers to straight or branched chain hydrocarbyl groups having from 1 to about 100 carbon atoms. Whenever it appears herein, a numerical range, such as "1 to 100" or "C1-C100", refers to each integer in the given range; e.g., "C1-C100 alkyl" means that an alkyl group may comprise only 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 100 carbon atoms, although the term "alkyl" also includes instances where no numerical range of carbon atoms is designated. For example, the term "alkyl" can refer to a sub-range between C1-C100 (e.g. C1-C6)."Substituted alkyl" refers to alkyl moieties bearing substituents including alkyl, alkenyl, alkynyl, hydroxy, oxo, alkoxy, mercapto, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted heterocyclic, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aryloxy, substituted aryloxy, halogen, haloalkyl, cyano, nitro, nitrone, amino, lower alkylamino, lower alkyldiamino, amido, azido, -C(O)H, -C(O)R9, -CH2OR9, -C(O)-, -S-, -S(O)2, -OC(O)-O-, wherein R9 is H or lower alkyl, acyl, oxyacyl, carboxyl, carbamate, sulfonyl, sulfonamide, sulfuryl, and the like. As used herein, "lower alkyl" refers to alkyl moieties having from 1 to about 6 carbon atoms.

[0014] As used herein, "alkenyl" refers to straight or branched chain hydrocarbyl groups having at least one carbon-carbon double bond, and having in the range of about 2 up to about 100 carbon atoms, and "substituted alkenyl" refers to alkenyl groups further bearing one or more substituents as set forth above. As used herein, "lower alkenyl" refers to alkenyl moieties having from 2 to about 6 carbon atoms.

[0015] As used herein, "alkynyl" refers to straight or branched chain hydrocarbyl groups having at least one carbon-carbon triple bond, and having in the range of about 2 up to about 100 carbon atoms, and "substituted alkynyl" refers to alkynyl groups further bearing one or more substituents as set forth above. As used herein, "lower alkynyl" refers to alkynyl moieties having from 2 to about 6 carbon atoms.

[0016] As used herein, "cycloalkyl" refers to cyclic (i.e., ring-containing) alkyl moieties typically containing in the range of about 3 up to about 8 carbon atoms, and "substituted cycloalkyl" refers to cycloalkyl groups further bearing one or more substituents as set forth above.

[0017] As used herein, "aryl" refers to aromatic groups having in the range of 5 up to 14 (e.g., phenyl) carbon atoms and "substituted aryl" refers to aryl groups further bearing one or more substituents as set forth above.

[0018] As used herein "arylene" and "heteroarylene" refer to an aryl ring or ring system or a heteroaryl ring or ring system which connects two other parts of a molecule, i.e. the two parts are bonded to the ring or ring system in two distinct ring positions. Arylene and heteroarylene may be substituted or unsubstituted. Unsubstituted arylene and heteroarylene have no substituents other than the two parts of the molecule it connects. Substituted arylene and heteroarylene have substituents in addition to the two parts of the molecule it connects.

[0019] An "effective amount" of a compound is an amount sufficient to contribute to the treatment, prevention (e.g. prophylaxis), or reduction of a symptom or symptoms of a disease. Where recited in reference to a disease treatment, an "effective amount" may also be referred to as a "therapeutically effective amount." A "reduction" of a symptom or symptoms (and grammatical equivalents of this phrase) means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s). A "prophylactically effective amount" of a drug is an amount of a drug that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) a disease, disorder or condition, or reducing the likelihood of the onset (or reoccurrence) of a disease, disorder or condition or symptoms thereof. The full prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations.

[0020] As used herein, "heteroaryl" refers to aromatic moieties containing one or more heteroatoms (e.g., N, O, S, or the like) as part of the ring structure and having in the range of 5 up to 14 total atoms in the ring structure (i.e., carbon atoms and heteroatoms). "Substituted heteroaryl" refers to heteroaryl groups further bearing one or more substituents as set forth above.

[0021] As used herein, "heterocyclic" refers to non-aromatic cyclic (i.e., ring-containing) groups containing one or more heteroatoms (e.g., N, O, S, or the like) as part of the ring structure, and having in the range of 3 up to 14 carbon atoms and "substituted heterocyclic" refers to heterocyclic groups further bearing one or more substituents as set forth above.

[0022] As used herein, "halogen" or "halide" refers to fluoride, chloride, bromide or iodide. "Fluoride, chloride, bromide or iodide" may also be referred to as "fluoro", "chloro", "bromo", or "iodo".

[0023] It will be readily apparent to those skilled in the art that some of the compounds of the invention may contain one or more asymmetric centers, such that the compounds may exist in enantiomeric as well as in diastereomeric forms. Unless it is specifically noted otherwise, the scope of the present invention includes all enantiomers, diastereomers and racemic mixtures. Some of the compounds of the invention may form salts with pharmaceutically acceptable acids or bases, and such pharmaceutically acceptable salts of the compounds described herein are also within the scope of the invention.

[0024] As used herein, "diastereomers" are stereoisomers in which asymmetric centers are different but which are not enantiomers.

[0025] In particular, a skilled person will realize that even if the absolute stereochemistry of a particular stereoisomer (e.g. an enantiomer or diastereomer) of a molecule is not known, that particular stereoisomer can be distinguished from the other stereoisomers by use of other techniques (e.g. polarimetry, nuclear magnetic resonance spectroscopy, chromatography, and others identifiable to a skilled person).

[0026] In particular, one exemplary method of distinguishing stereoisomers when the absolute stereochemistry of each stereoisomer is not known is chromatography such as high pressure liquid chromatography (HPLC) or thin-layer chromatograph (TLC). In particular, two or more stereoisomers such as diastereomers can be separated and characterized by their retention times and of Rf values, which would be expected to be replicable by using the same chromatographic conditions (e.g. flow rate, column material, TLC stationary phase, solvent systems/gradient profiles, and others identifiable to a skilled person). In particular, a skilled person will realize that even when the exact relative retention times and/or Rf values of one or more stereoisomers is not replicated (e.g. due to slight variations in the chromatographic parameters and/or chromatographic equipment), a stereoisomer with a shorter retention time can be said to be "faster eluting" and a stereoisomer with a linger retention time can be said to be "slower eluting", and similarly, a stereoisomer with smaller Rf value can be said to have a "low Rf" and a stereoisomer with a larger Rf value can be said to have a "high Rf".

[0027] A skilled person will realize that once two or more stereoisomers are distinguished by a technique such as chromatography (e.g. HPLC and/or TLC), the absolute stereochemistry of the stereoisomers can be determined by techniques or combinations of techniques identifiable to a skilled person (e.g. x-ray crystallography, vibrational circular dichroism, nuclear magnetic resonance, total synthesis, and others identifiable to a skilled person).

[0028] A "pharmaceutically acceptable salt" is any salt that retains the activity of the parent compound and does not impart any additional deleterious or untoward effects on the subject to which it is administered and in the context in which it is administered compared to the parent compound. A pharmaceutically acceptable salt also refers to any salt which may form in vivo as a result of administration of an acid, another salt, or a prodrug which is converted into an acid or salt.

[0029] Pharmaceutically acceptable salts of acidic functional groups may be derived from organic or inorganic bases. The salt may comprise a mono or polyvalent ion. Of particular interest are the inorganic ions, lithium, sodium, potassium, calcium, and magnesium. Organic salts may be made with amines, particularly ammonium salts such as mono-, di- and trialkyl amines or ethanol amines. Salts may also be formed with caffeine, tromethamine and similar molecules. Hydrochloric acid or some other pharmaceutically acceptable acid may form a salt with a compound that includes a basic group, such as an amine or a pyridine ring. The term "pharmaceutically acceptable salt" is also meant to include salts of active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66:1-19). Certain specific compounds of the present invention may contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0030] The terms "treating" or "treatment" refers to any indicia of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a patient's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination.

EXAMPLES



[0031] The following examples are intended only to illustrate the invention and should in no way be construed as limiting the invention.

SYNTHETIC PROCEDURES



[0032] Scheme 1 sets forth below outlines a synthetic route to the compound of the invention described in Examples 1 and 2:





[0033] Scheme 2 set forth below outlines a synthetic route to the compounds described in Examples 3-6 and 8-35:





[0034] The following scheme (Scheme 3) sets forth a synthetic route to the compounds of the invention described in Example 7:



[0035] Scheme 4 set forth below outlines a synthetic route to the compound described in Examples 36-39:




Example 1


4-((2-(3,5-Dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer):



[0036] 


Step 1. Ethyl 2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxylate



[0037] Palladium acetate (3.3 mg, 0.015 mmol) and SPhos (11.8 mg, 0.029 mmol) were added to a solution of ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclopent-1-enecarboxylate (Combiblocks, Inc., 411 mg, 1.43 mmol) in THF (1.43 mL). The reaction mixture was purged with nitrogen and a solution of 3,5-dimethoxybenzylzinc chloride (Aldrich, 3.4 mL of a 0.5 M solution in THF, 1.7 mmol) was added drop-wise. After stirring at room temperature overnight, the reaction mixture was quenched with saturated aqueous NH4Cl (25 mL) and extracted with EtOAc (3x75 mL). The combined extracts were dried (Na2SO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (40 g gold column, hexanes → 50% EtOAc/hexanes, gradient), to afford 353 mg (85%) of ethyl 2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxylate.

Step 2. Ethyl 2-(3,5-dimethoxybenzyl)cyclopentanecarboxylate



[0038] Palladium on carbon (10 wt%, 5 mg) was added to a solution of ethyl 2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxylate (100 mg, 0.34 mmol) in methanol (3.4 mL). A hydrogen atmosphere was established by evacuating and refilling with hydrogen (3x). The reaction mixture placed in a 40 °C bath and stirred under a balloon of hydrogen overnight. The mixture was then cooled and filtered through celite, washing with excess methanol. The filtrate was concentrated in vacuo to afford 100 mg (quant.) of ethyl 2-(3,5-dimethoxybenzyl)cyclopentanecarboxylate.

Step 3. 2-(3,5-Dimethoxybenzyl)cyclopentanecarboxylic acid



[0039] Aqueous lithium hydroxide (1.7 mL of a 1.0 M solution, 1.7 mmol) was added to a solution of ethyl 2-(3,5-dimethoxybenzyl)cyclopentanecarboxylate (100 mg, 0.34 mmoL) in THF (3.4 mL) in a scintillation vial. The vial was heated at 60 °C for 3 days then cooled to room temperature. The organic solvent was removed under a stream of nitrogen, then the residue was diluted with water (5 mL), acidified with 1 N HCl (2 mL) and extracted with EtOAc (3x20 mL). The extracts were dried (Na2SO4), filtered and concentrated to afford 98 mg (quant.) of 2-(3,5-dimethoxybenzyl)cyclopentanecarboxylic acid.

Step 4. Methyl 4-((2-(3,5-dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoate



[0040] Triethylamine (28 µL, 0.20 mmol), DMAP (56 mg, 0.45 mmol), methyl 4-(aminomethyl)benzoate hydrochloride (41 mg, 0.20 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (50 mg, 0.26 mmol) were added sequentially to a solution of 2-(3,5-dimethoxybenzyl)cyclopentanecarboxylic acid (52 mg, 0.20 mmol) in CH2Cl2 (2.0 mL). The heterogeneous mixture was allowed to stir at room temperature overnight, during which time the reaction became homogeneous. The reaction solution was treated with saturated aqueous NH4Cl (10 mL) and extracted with EtOAc (3x20 mL). The combined extracts were dried (Na2SO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (4 g gold column, hexanes → EtOAc, gradient), to afford 13 mg (16%) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoate (high Rf diastereomer) and 3.6 mg (4%) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 5. 4-((2-(3,5-Dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0041] Aqueous lithium hydroxide (0.15 mL of a 1.0 M solution, 0.15 mmol) was added to a solution of methyl 4-((2-(3,5-dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoate (high Rf diastereomer, 13 mg, 0.032 mmol) in THF (0.3 mL) in a 1-dram vial. The vial was heated at 50 °C overnight then cooled to room temperature. The organic solvent was removed under a stream of nitrogen, then the residue was diluted with water (0.5 mL), acidified with 1 N HCl (0.5 mL) and extracted with EtOAc (3x2 mL). The extracts were dried (Na2SO4), filtered and concentrated to afford 11 mg (88%) of the title compound.

Example 2


4-((2-(3,5-Dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 1):



[0042] 



[0043] In accordance with the procedure of Example 1, step 5, methyl 4-((2-(3,5-dimethoxybenzyl)cyclopentanecarboxamido)methyl)benzoate (low Rf diastereomer, 3.6 mg, 0.009 mmol) was converted into 3 mg (86%) of the title compound.

Example 3


4-((2-(3,5-Dimethoxybenzyl)cyclopent-1-enecarboxamido)methyl)benzoic acid:



[0044] 


Step 1. 2-(3,5-Dimethoxybenzyl)cyclopent-1-enecarboxylic acid



[0045] Aqueous lithium hydroxide (4.1 mL of a 1.0 M solution, 4.1 mmol) was added to a solution of ethyl 2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxylate (68.7 mg, 0.237 mmol) in THF (4.1 mL). The vial was heated at 60 °C for 24 h after which time TLC analysis showed little if any reaction had occurred. A second portion of lithium hydroxide (351 mg, 8.20 mmol) was added, and the mixture was stirred for an additional 18 h at 60 °C. The mixture was then cooled to room temperature, acidified with 1 N HCl and extracted with CH2Cl2 (3x). The extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (4 g gold column, 5% EtOAc/hexanes → 60% EtOAc/hexanes, gradient), to afford 22.5 mg (36%) of 2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxylic acid.

Step 2. Methyl 4-((2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxamido) methyl)benzoate



[0046] TBTU (O-Benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorborate (28.0 mg, 0.0863 mmol), methyl 4-(aminomethyl)benzoate hydrochloride (18.1 mg, 0.0870 mmol) and diisopropylethylamine (45 µL, 0.26 mmol), were added to a solution of 2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxylic acid (22.5 mg, 0.858 mmol) in DMF (0.43 mmol). After stirring at 40 °C for 18 h, the reaction mixture was diluted with EtOAc and washed with 1 N HCl (2x), 1 N NaOH (2x) and brine. The organic phase was dried (MgSO4), filtered and concentrated in vacuo to afford 35 mg (99%) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxamido)methyl)benzoate as a white solid.

Step 3. 4-((2-(3,5-Dimethoxybenzyl)cyclopent-1-enecarboxamido)methyl)benzoic acid



[0047] Water (363 µL) and aqueous lithium hydroxide (0.56 mL of a 1.0 M solution, 0.56 mL) were added to a solution of methyl 4-((2-(3,5-dimethoxybenzyl)cyclopent-1-enecarboxamido)methyl)benzoate (11.0 mg, 0.0269 mmol) in THF (0.75 mL). After stirring at 30 °C for 18 h, the mixture was cooled, acidified with 1 N HCl and extracted with CH2Cl2 (3x). The combined extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (4 g gold column, 30% EtOAc/hexanes → 60% EtOAc/hexanes (both solvents containing 0.5% AcOH), gradient), to afford 5.9 mg (55%) of the title compound.

Example 4


4-((2-(3-(Trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoic acid:



[0048] 


Step 1. Ethyl 2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxylate



[0049] Palladium acetate (3.1 mg, 0.014 mmol) and SPhos (10.9 mg, 0.0258 mmol) were added to a solution of ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclopent-1-enecarboxylate (Combiblocks, Inc., 380 mg, 1.25 mmol) in THF (1.3 mL). The reaction mixture was purged with nitrogen for 10 min and a solution of 3-trifluormethylbenzylzinc chloride (Aldrich, 3.1 mL of a 0.5 M solution in THF, 1.6 mmol) was added drop-wise. After stirring at room temperature overnight, the reaction mixture was quenched with saturated aqueous NH4Cl and extracted with EtOAc (3x). The combined extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (40 g gold column, hexanes → 15% EtOAc/hexanes, gradient), to afford 342 mg (92%) of ethyl 2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxylate.

Step 2. 2-(3-(Trifluoromethyl)benzyl)cyclopent-1-enecarboxylic acid



[0050] Aqueous lithium hydroxide (6.5 mL of a 3.0 M solution, 20 mmol) was added to a solution of ethyl 2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxylate (111 mg, 0.373 mmol) in 1,4-dioxane (6.5 mL). The reaction mixture was heated at 70 °C for 18 h then cooled to room temperature. The reaction mixture was acidified with 1 N HCl and extracted with CH2Cl2 (3x). The extracts were dried (MgSO4), filtered and concentrated in vacuo. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (12 g gold column, 5% EtOAc/hexanes → 40% EtOAc/hexanes, gradient), to afford 83.1 mg (83%) of 2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxylic acid.

Step 3. Methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido) methyl)benzoate



[0051] In accordance with the procedure of Example 3, step 2, 2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxylic acid (81.7 mg, 0.302 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (63.1 mg, 0.304 mmol) were converted into 126 mg (quant.) of methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(3-(Trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoic acid



[0052] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate (10.4 mg, 0.0249 mmol) was converted into 6.8 mg (68%) of the title compound.

Example 5


4-((2-(3-(Trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer, see Example 4):



[0053] 


Step 1.Methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoate



[0054] Palladium on carbon (10 wt%, 8.7 mg) was added to a solution of methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate (47.9 mg, 0.115 mmol) in ethanol (1.7 mL). A hydrogen atmosphere was established by evacuating and refilling with hydrogen (3x). The reaction mixture placed in a 40 °C bath and stirred under a balloon of hydrogen overnight. The mixture was then cooled and filtered through celite, washing with excess ethanol and the filtrate was concentrated in vacuo. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (4 g gold column, 20% EtOAc/hexanes → 50% EtOAc/hexanes, gradient), to afford 30.5 mg (63%) of methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoate (high Rf diastereomer) and 13.8 mg (29%) of methyl 4-((2-(3-(trifluoromethyl)benzyl) cyclopentanecarboxamido) methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(3-(Trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0055] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoate (high Rf diastereomer, 17.2 mg, 0.0410 mmol) was converted into 8.2 mg (49%) of the title compound.

Example 6


4-((2-(3-(Trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 5):



[0056] 



[0057] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopentanecarboxamido)methyl)benzoate (low Rf diastereomer, 5.9 mg, 0.014 mmol) was converted into 2.6 mg (46%) of the title compound.

Example 7


4-((N-Methyl-2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoic acid:



[0058] 


Step 1. Methyl 4-((N-methyl-2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate



[0059] A solution of lithium diisopropylamide (0.09 mL of a 2.0 M solution in THF/heptane/ethylbenzene, 0.2 mmol) was added to a - 78 °C solution of methyl 4-((2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate (see Example 4, 64.5 mg, 0.155 mmol) in THF (1.0 mL). After 20 min at -78 °C, iodomethane (0.10 mL of a 2.0 M solution in t-butyl ether, 0.20 mmol) was added. The mixture was allowed to warm to room temperature and stirred for 18 h. The reaction was quenched with saturated aqueous NH4Cl and extracted with CH2Cl2 (3x). The combined extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (12 g gold column, 20% EtOAc/hexanes → 50% EtOAc/hexanes, gradient), to afford 46.6 mg (70%) of methyl 4-((N-methyl-2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate, which appears as a 3:2 ratio of amide rotamers.

Step 2. 4-((N-Methyl-2-(3-(triffuoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoic acid



[0060] In accordance with the procedure of Example 3, step 3, of methyl 4-((N-methyl-2-(3-(trifluoromethyl)benzyl)cyclopent-1-enecarboxamido)methyl)benzoate (10.0 mg, 0.0232 mmol) was converted into 4.8 mg (50%) of the title compound.

Example 8


4-((2-(3,5-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid:



[0061] 


Step 1. Ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate



[0062] Sodium hydride (60 wt. % in oil, 228 mg, 5.70 mmol) was added to a 0 °C solution of ethyl 2-oxocyclohexanecarboxylate (888 mg, 4.96 mmol) in THF (25 mL). After 40 min at 0 °C, N-phenyl-bis(trifluoromethanesulfonimide) (2.14 g, 5.93 mmol) was added and the solution was allowed to warm to room temperature and stir overnight. The reaction mixture was quenched with saturated aqueous NaHCO3 and extracted with CH2Cl2 (3x). The combined extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (120 g gold column, hexanes → 25% EtOAc/hexanes, gradient), to afford 879 mg (59%) of ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate.

Step 2. Ethyl 2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxylate



[0063] Palladium acetate (2.8 mg, 0.012 mmol) and SPhos (10.1 mg, 0.0239 mmol) were added to a solution of ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (293 mg, 0.967 mmol) in THF (1.0 mL). The reaction mixture was purged with nitrogen for 10 min and a solution of 3,5-dimethoxybenzylzinc chloride (Aldrich, 2.3 mL of a 0.5 M solution in THF, 1.2 mmol) was added drop-wise. After stirring at room temperature overnight, the reaction mixture was quenched with saturated aqueous NH4Cl and extracted with EtOAc (3x). The combined extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (24 g gold column, hexanes → 25% EtOAc/hexanes, gradient), to afford 194 mg (66%) of ethyl 2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxylate.

Step 3. 2-(3,5-Dimethoxybenzyl)cyclohex-1-enecarboxylic acid



[0064] In accordance with the procedure of Example 4, step 2, ethyl 2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxylate (189 mg, 0.619 mmol) was converted into 122 mg (72%) of 2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxylic acid.

Step 4. Methyl 4-((2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0065] In accordance with the procedure of Example 3, step 2, 2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxylic acid (120 mg, 0.435 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (90.6 mg, 0.436 mmol) were converted into 188 mg (quant.) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate.

Step 5. 4-((2-(3,5-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0066] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (11.4 mg, 0.0269 mmol) was converted into 5.0 mg (45%) of the title compound.

Example 9


4-((2-(3,5-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer):



[0067] 


Step 1. Methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate



[0068] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(3,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (42.9 mg, 0.101 mmol) was converted into 25.0 mg (58%) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 14.0 mg (33%) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(3,5-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0069] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 24.3 mg, 0.0571 mmol) was converted into 14.8 mg (63%) of the title compound.

Stereochemical Assignments



[0070] The two diastereomers isolated in Example 9, step 1 [methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer)] were examined in a ROESY NMR experiment in order to determine their relative stereochemistry. Based on the enhancements observed, it was determined that the high Rf diastereomer has a cis arrangement of the groups appended to the cyclohexane core, and the low Rf diastereomer has a trans arrangement of the groups appended to the cyclohexane core (see Figure 1 below). This assignment is expected to hold for all the sets of diastereomers described herein, however a skilled person can determine the relative and/or absolute stereochemistry by using routine techniques known in the art (e.g. ROESY NMR).


Example 10


4-((2-(3,5-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 9):



[0071] 



[0072] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 13.7 mg, 0.0322 mmol) was converted into 5.8 mg (44%) of the title compound.

Example 11


4-((2-(3,5-Dimethoxybenzyl)cyclohept-1-enecarboxamido)methyl)benzoic acid:



[0073] 


Step 1. Methyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohept-1-enecarboxylate



[0074] In accordance with the procedure of Example 8, step 1, methyl 2-oxocycloheptanecarboxylate (797 mg, 4.64 mmol) was converted into 621 mg (44%) of methyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohept-1-enecarboxylate after purification on a Teledyne-Isco Combiflash machine (80 g gold column, hexanes → 25% EtOAc/hexanes, gradient).

Step 2. Methyl 2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxylate



[0075] In accordance with the procedure of Example 8, step 2, methyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohept-1-enecarboxylate (256 mg, 0.846 mmol) was converted into 167 mg (65%) of methyl 2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxylate.

Step 3. 2-(3,5-Dimethoxybenzyl)cyclohept-1-enecarboxylic acid



[0076] In accordance with the procedure of Example 4, step 2, methyl 2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxylate (167 mg, 0.549 mmol) was converted into 137 mg (86%) of 2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxylic acid.

Step 4. Methyl 4-((2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxamido)methyl)benzoate



[0077] In accordance with the procedure of Example 3, step 2, 2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxylic acid (124 mg, 0.427 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (89.2 mg, 0.429 mmol) were converted into 189 mg (quant.) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxamido)methyl)benzoate.

Step 5. 4-((2-(3,5-Dimethoxybenzyl)cyclohept-1-enecarboxamido)methyl)benzoic acid



[0078] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxamido)methyl)benzoate (8.6 mg, 0.020 mmol) was converted into 2.3 mg (28%) of the title compound.

Example 12


4-((2-(3,5-Dimethoxybenzyl)cycloheptanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer, see Example 11):



[0079] 


Step 1. Methyl 4-((2-(3,5-dimethoxybenzyl)cycloheptanecarboxamido) methyl)benzoate



[0080] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(3,5-dimethoxybenzyl)cyclohept-1-enecarboxamido)methyl)benzoate (41.6 mg, 0.0951 mmol) was converted into 25.0 mg (62%) of methyl 4-((2-(3,5-dimethoxybenzyl)cycloheptanecarboxamido)methyl) benzoate (high Rf diastereomer) and 9.6 mg (23%) of methyl 4-((2-(3,5-dimethoxybenzyl)cycloheptanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(3,5-Dimethoxybenzyl)cycloheptanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0081] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3,5-dimethoxybenzyl)cycloheptanecarboxamido)methyl)benzoate (high Rf diastereomer, 23.8 mg, 0.0541 mmol) was converted into 15.3 mg (67%) of the title compound.

Example 13


4-((2-(3,5-Dimethoxybenzyl)cycloheptanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 12):



[0082] 



[0083] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3,5-dimethoxybenzyl)cycloheptanecarboxamido)methyl)benzoate (low Rf diastereomer, 9.5 mg, 0.022 mmol) was converted into 3.3 mg (36%) of the title compound.

Example 14


4-((2-(2,3-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid:



[0084] 


Step 1. Ethyl 2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxylate



[0085] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (193 mg, 0.64 mmol) and 2,3-dimethoxybenzylzinc chloride (Rieke Metals, 1.5 mL of a 0.5 M solution in THF, 0.75 mmol) were converted into 123 mg (63%) of ethyl 2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxylate after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 25% EtOAc/hexanes, gradient).

Step 2. 2-(2,3-Dimethoxybenzyl)cyclohex-1-enecarboxylic acid



[0086] In accordance with the procedure of Example 4, step 2, ethyl 2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxylate (120 mg, 0.395 mmol) was converted into 66.6 mg (61%) of 2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxylic acid.

Step 3. Methyl 4-((2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0087] In accordance with the procedure of Example 3, step 2, 2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxylic acid (64.8 mg, 0.234 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (49 mg, 0.235 mmol) were converted into 103 mg (quant.) of methyl 4-((2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0088] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(2,3-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (12.0 mg, 0.0283 mmol) was converted into 5.4 mg (47%) of the title compound.

Example 15


4-((2-(2,3-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer):



[0089] 


Step 1. Methyl 4-((2-(2,3-dimethoxybenzyl)cyclohexanecarboxamido) methyl)benzoate



[0090] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(2,3-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (38.8 mg, 0.0916 mmol) was converted into 20.1 mg (52%) of methyl 4-((2-(2,3-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 15.0 mg (39%) of methyl 4-((2-(2,3-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(2,3-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0091] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2,3-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 20.1 mg, 0.0472 mmol) was converted into 11.2 mg (58%) of the title compound.

Example 16


4-((2-(2,3-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 14):



[0092] 



[0093] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2,3-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 15.0 mg, 0.0353 mmol) was converted into 6.4 mg (44%) of the title compound.

Example 17


4-((2-(2,5-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid:



[0094] 


Step 1. Ethyl 2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxylate



[0095] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (193 mg, 0.637 mmol) and 2,5-dimethoxybenzylzinc chloride (Rieke Metals, 1.5 mL of a 0.5 M solution in THF, 0.75 mmol) were converted into 142 mg (73%) of ethyl 2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxylate after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 25% EtOAc/hexanes, gradient).

Step 2. 2-(2,5-Dimethoxybenzyl)cyclohex-1-enecarboxylic acid



[0096] In accordance with the procedure of Example 4, step 2, ethyl 2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxylate (137 mg, 0.450 mmol) was converted into 73.5 mg (59%) of 2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxylic acid.

Step 3. Methyl 4-((2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxamido) methyl)benzoate



[0097] In accordance with the procedure of Example 3, step 2, 2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxylic acid (71.9 mg, 0.260 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (54.0 mg, 0.260 mmol) were converted into 110 mg (quant.) of methyl 4-((2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(2,5-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0098] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (10.3 mg, 0.0243 mmol) was converted into 5.8 mg (58%) of the title compound.

Example 18


4-((2-(2,5-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer):



[0099] 


Step 1. Methyl 4-((2-(2,5-dimethoxybenzyl)cyclohexanecarboxamido) methyl)benzoate



[0100] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(2,5-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (40.3 mg, 0.0952 mmol) was converted into 21.0 mg (52%) of methyl 4-((2-(2,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 13.8 mg (34%) of methyl 4-((2-(2,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(2,5-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0101] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 20.0 mg, 0.0470 mmol) was converted into 6.7 mg (35%) of the title compound.

Example 19


4-((2-(2,5-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 18):



[0102] 



[0103] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2,5-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 13.8 mg, 0.0324 mmol) was converted into 5.4 mg (41%) of the title compound.

Example 20


4-((2-(3,5-Dimethoxybenzyl)cyclooct-1-enecarboxamido)methyl)benzoic acid:



[0104] 


Step 1. Ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclooct-1-enecarboxylate



[0105] In accordance with the procedure of Example 8, step 1, ethyl 2-oxocyclooctanecarboxylate (Aldrich, 829 mg, 4.06 mmol) was converted into 497 mg (37%) of ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclooct-1-enecarboxylate.

Step 2. Ethyl 2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxylate



[0106] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (224 mg, 0.679 mmol) and of 3,5-dimethoxybenzylzinc chloride (Aldrich, 1.6 mL of a 0.5 M solution in THF, 0.80 mmol) were converted into 98.4 mg (44%) of ethyl 2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxylate.

Step 3. 2-(3,5-Dimethoxybenzyl)cyclooct-1-enecarboxylic acid



[0107] In accordance with the procedure of Example 4, step 2, ethyl 2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxylate (94.8 mg, 0.285 mmol), with stirring at 80 °C, was converted into 20.7 mg (24%) of 2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxylic acid, along with a 60.0 mg (63%) of starting ester.

Step 4. Methyl 4-((2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxamido)methyl)benzoate



[0108] In accordance with the procedure of Example 3, step 2, 2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxylic acid (62.3 mg, 0.205 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (42.7 mg, 0.205 mmol) were converted into 85.8 mg (93%) of methyl 4-((2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxamido)methyl)benzoate.

Step 5. 4-((2-(3,5-Dimethoxybenzyl)cyclooct-1-enecarboxamido)methyl)benzoic acid



[0109] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(3,5-dimethoxybenzyl)cyclooct-1-enecarboxamido)methyl)benzoate (11.1 mg, 0.0246 mmol) was converted into 5.7 mg (53%) of the title compound.

Example 21


4-((2-(3,4-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0110] 


Step 1. Ethyl 2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxylate



[0111] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (198 mg, 0.654 mmol) and 3,4-dimethoxybenzylzinc chloride (Rieke Metals, 1.6 mL of a 0.5 M solution in THF, 0.80 mmol) were converted into 122 mg (61%) of ethyl 2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxylate after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 20% EtOAc/hexanes, gradient).

Step 2. 2-(3,4-Dimethoxybenzyl)cyclohex-1-enecarboxylic acid



[0112] In accordance with the procedure of Example 4, step 2, ethyl 2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxylate (120 mg, 0.393 mmol) was converted into 92 mg (85%) of 2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxylic acid.

Step 3. Methyl 4-((2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0113] In accordance with the procedure of Example 3, step 2, 2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxylic acid (92 mg, 0.33 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (69 mg, 0.33 mmol) were converted into 131 mg (93%) of methyl 4-((2-(3,4-dimethoxybenzy1)cyclohex-1-enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(3,4-Dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0114] In accordance with the procedure of Example 3, step 3, of methyl 4-((2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (11.2 mg, 0.0264 mmol) was converted into 4.3 mg (40%) of the title compound.

Example 22


4-((2-(3,4-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0115] 


Step 1. Methyl 4-((2-(3,4-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate



[0116] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(3,4-dimethoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (40 mg, 0.095 mmol) was converted into 27.5 mg (68%) of methyl 4-((2-(3,4-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 10.9 mg (27%) of methyl 4-((2-(3,4-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(3,4-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0117] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3,4-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 27.5 mg, 0.0646 mmol) was converted into 16.1 mg (61%) of the title compound.

Example 23


4-((2-(3,4-Dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer, see Example 22)



[0118] 



[0119] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3,4-dimethoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 10.4 mg, 0.0244 mmol) was converted into 4.1 mg (41%) of the title compound.

Example 24


4-((2-(2-Chlorobenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0120] 


Step 1. Ethyl 2-(2-chlorobenzyl)cyclohex-1-enecarboxylate



[0121] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (300 mg, 0.993 mmol) and 2-chlorobenzylzinc(II) chloride (Aldrich, 2.4 mL of a 0.5 M solution in THF, 1.2 mmol) were converted into 144 mg (52%) of ethyl 2-(2-chlorobenzyl)cyclohex-1-enecarboxylate after purification on a Teledyne-Isco Combiflash machine (24 g gold column, hexanes → 15% EtOAc/hexanes, gradient).

Step 2. 2-(2-Chlorobenzyl)cyclohex-1-enecarboxylic acid



[0122] In accordance with the procedure of Example 4, step 2, ethyl 2-(2-chlorobenzyl)cyclohex-1-enecarboxylate (55 mg, 0.197 mmol) was converted into 42.4 mg (86%) of 2-(2-chlorobenzyl)cyclohex-1-enecarboxylic acid after purification on a Teledyne-Isco Combiflash machine (4 g gold column, hexanes → 35% EtOAc/hexanes, gradient).

Step 3. Methyl 4-((2-(2-chlorobenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0123] In accordance with the procedure of Example 3, step 2, 2-(2-chlorobenzyl)cyclohex-1-enecarboxylic acid (41 mg, 0.16 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (34 mg, 0.16 mmol) were converted into 64 mg (99%) of methyl 4-((2-(2-chlorobenzyl)cyclohex-1-enecarboxamido)methyl)benzo ate.

Step 4. 4-((2-(2-Chlorobenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0124] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2-chlorobenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (9.4 mg, 0.0236 mmol) was converted into 4.0 mg (44%) of the title compound.

Example 25


4-((2-Benzylcyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0125] 


Step 1. Methyl 4-((2-benzylcyclohexanecarboxamido)methyl)benzoate



[0126] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(2-chlorobenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (47 mg, 0.12 mmol) was converted into 26.6 mg (62%) of methyl 4-((2-benzylcyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 9.3 mg (22%) of methyl 4-((2-benzylcyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer). No product containing the starting chloro substituent was isolated.

Step 2. 4-((2-Benzylcyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0127] In accordance with the procedure of Example 3, step 3, methyl 4-((2-benzylcyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 26.6 mg, 0.0728 mmol) was converted into 18.7 mg (73%) of the title compound.

Example 26


4-((2-Benzylcyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer)



[0128] 



[0129] In accordance with the procedure of Example 3, step 3, methyl 4-((2-benzylcyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 9.3 mg, 0.0254 mmol) was converted into 4.5 mg (50%) of the title compound.

Example 27


4-((2-(3-Methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0130] 


Step 1. Ethyl 2-(3-methoxybenzyl)cyclohex-1-enecarboxylate



[0131] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (190 mg, 0.628 mmol) and 3-methoxybenzylzinc(II) chloride (Rieke Metals, 1.5 mL of a 0.5 M solution in THF, 0.75 mmol) were converted into 152 mg of ethyl 2-(3-methoxybenzyl)cyclohex-1-enecarboxylate contaminated with the triflate starting material after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 15% EtOAc/hexanes, gradient). The estimated yield of the desired product, accounting for impurity based on 1H-NMR analysis, is approximately 115 mg (∼ 67%).

Step 2. 2-(3-Methoxybenzyl)cyclohex-1-enecarboxylic acid



[0132] In accordance with the procedure of Example 4, step 2, ethyl 2-(3-methoxybenzyl)cyclohex-1-enecarboxylate (152 mg of the impure material from step 1, ∼ 0.42 mmol) was converted into 86.4 mg (- 84%) of 2-(3-methoxybenzyl)cyclohex-1-enecarboxylic acid after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 35% EtOAc/hexanes, gradient).

Step 3. Methyl 4-((2-(3-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0133] In accordance with the procedure of Example 3, step 2, 2-(3-methoxybenzyl)cyclohex-1-enecarboxylic acid (83 mg, 0.34 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (70 mg, 0.34 mmol) were converted into 136 mg (quant.) of methyl 4-((2-(3-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(3-Methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0134] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (8.3 mg, 0.021 mmol) was converted into 2.9 mg (36%) of the title compound.

Example 28


4-((2-(3-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0135] 


Step 1. Methyl 4-((2-(3-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate



[0136] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(3-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (37 mg, 0.094 mmol) was converted into 15 mg (40%) of methyl 4-((2-(3-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 7.2 mg (19%) of methyl 4-((2-(3-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(3-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0137] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 15 mg, 0.038 mmol) was converted into 8.2 mg (57%) of the title compound.

Example 29


4-((2-(3-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer)



[0138] 



[0139] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(3-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 7.2 mg, 0.018 mmol) was converted into 2.9 mg (42%) of the title compound.

Example 30


4-((2-(2-Methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0140] 


Step 1. Ethyl 2-(2-methoxybenzyl)cyclohex-1-enecarboxylate



[0141] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (190 mg, 0.628 mmol) and 2-methoxybenzylzinc(II) chloride (Rieke Metals, 1.5 mL of a 0.5 M solution in THF, 0.75 mmol) were converted into 160 mg of ethyl 2-(2-methoxybenzyl)cyclohex-1-enecarboxylate contaminated with the triflate starting material after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 15% EtOAc/hexanes, gradient). The estimated yield of the desired product, accounting for impurity based on 1H-NMR analysis, is approximately 119 mg (∼69%).

Step 2. 2-(2-Methoxybenzyl)cyclohex-1-enecarboxylic acid



[0142] In accordance with the procedure of Example 4, step 2, ethyl 2-(2-methoxybenzyl)cyclohex-1-enecarboxylate (160 mg of the impure material from step 1, ∼ 0.42 mmol) was converted into 84 mg (∼ 81%) of 2-(2-methoxybenzyl)cyclohex-1-enecarboxylic acid after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 35% EtOAc/hexanes, gradient).

Step 3. Methyl 4-((2-(2-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0143] In accordance with the procedure of Example 3, step 2, 2-(2-methoxybenzyl)cyclohex-l-enecarboxylic acid (80 mg, 0.33 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (68 mg, 0.33 mmol) were converted into 121 mg (94%) of methyl 4-((2-(2-methoxybenzyl)cyclohex-1 -enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(2-Methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0144] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (10.3 mg, 0.0262 mmol) was converted into 4.5 mg (45%) of the title compound.

Example 31


4-((2-(2-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0145] 


Step 1. Methyl 4-((2-(2-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate



[0146] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(2-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (40 mg, 0.10 mmol) was converted into 18.8 mg (47%) of methyl 4-((2-(2-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 13.5 mg (34%) of methyl 4-((2-(2-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(2-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0147] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 18.8 mg, 0.0475 mmol) was converted into 8.9 mg (49%) of the title compound.

Example 32


4-((2-(2-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer)



[0148] 



[0149] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(2-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 13.5 mg, 0.0341 mmol) was converted into 7.2 mg (55%) of the title compound.

Example 33


4-((2-(4-Methoxybenzyl)cyc|ohex-1-enecarboxamido)methyl)benzoic acid



[0150] 


Step 1. Ethyl 2-(4-methoxybenzyl)cyclohex-1-enecarboxylate



[0151] In accordance with the procedure of Example 8, step 2, ethyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohex-1-enecarboxylate (199 mg, 0.659 mmol) and 4-methoxybenzylzinc(II) chloride (Rieke Metals, 1.6 mL of a 0.5 M solution in THF, 0.80 mmol) were converted into 156 mg of ethyl 2-(4-methoxybenzyl)cyclohex-1-enecarboxylate contaminated with the triflate starting material after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 15% EtOAc/hexanes, gradient). The estimated yield of the desired product, accounting for impurity based on 1H-NMR analysis, is approximately 135 mg (∼ 75%).

Step 2. 2-(4-Methoxybenzyl)cyclohex-1-enecarboxylic acid



[0152] In accordance with the procedure of Example 4, step 2, ethyl 2-(4-methoxybenzyl)cyclohex-1-enecarboxylate (156 mg of the impure material from step 1, ∼ 0.49 mmol) was converted into 98 mg (∼81%) of 2-(4-methoxybenzyl)cyclohex-1-enecarboxylic acid after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 35% EtOAc/hexanes, gradient).

Step 3. Methyl 4-((2-(4-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate



[0153] In accordance with the procedure of Example 3, step 2, 2-(4-methoxybenzyl)cyclohex-1-enecarboxylic acid (96.3 mg, 0.39 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (81.3 mg, 0.39 mmol) were converted into 153 mg (99%) of methyl 4-((2-(4-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate.

Step 4. 4-((2-(4-Methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoic acid



[0154] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(4-methoxybenzyl)cyclohex-1-enecarboxamido)methyl)benzoate (10.6 mg, 0.0269 mmol) was converted into 5.6 mg (55%) of the title compound.

Example 34


4-((2-(4-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0155] 


Step 1. Methyl 4-((2-(4-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate



[0156] In accordance with the procedure of Example 5, step 1, methyl 4-((2-(4-methoxybenzyl)cyclohex-l-enecarboxamido)methyl)benzoate (40 mg, 0.10 mmol) was converted into 26.1 mg (65%) of methyl 4-((2-(4-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer) and 10.1 mg (25%) of methyl 4-((2-(4-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer).

Step 2. 4-((2-(4-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0157] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(4-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (high Rf diastereomer, 25.7 mg, 0.065 mmol) was converted into 15.9 mg (64%) of the title compound.

Example 35


4-((2-(4-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from low Rf ester diastereomer)



[0158] 



[0159] In accordance with the procedure of Example 3, step 3, methyl 4-((2-(4-methoxybenzyl)cyclohexanecarboxamido)methyl)benzoate (low Rf diastereomer, 9.8 mg, 0.0248 mmol) was converted into 3.2 mg (34%) of the title compound.

Example 36


4-((3-(3,5-Dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxamido)methyl)benzoic acid



[0160] 


Step 1. Methyl 3-oxobicyclo[2.2.1]heptane-2-carboxylate



[0161] A solution of lithium diisopropylamide (2.3 mL of a 2.0 M solution in THF, 4.6 mmol) was added to a - 78 °C solution of norcamphor (Aldrich, 459 mg, 4.08 mmol) in THF (23.5 mL). After 15 min at - 78 °C, a solution of methyl cyanoformate (421 mg, 4.90 mmol) in THF (1 mL + 1 mL rinse) was added. The mixture was allowed to warm to room temperature and stirred for 18 h. The reaction was quenched with saturated aqueous NH4Cl and extracted with CH2Cl2 (3x). The combined extracts were dried (MgSO4), filtered and concentrated. The resulting crude residue was purified on a Teledyne-Isco Combiflash machine (40 g gold column, 10% EtOAc/hexanes → 30% EtOAc/hexanes, gradient), to afford 572 mg (83%) of methyl 3-oxobicyclo[2.2.1]heptane-2-carboxylate.

Step 2. Methyl 3-(((trifluoromethyl)sulfonyl)oxy)bicyclo[2.2.1]hept-2-ene-2-carboxylate



[0162] In accordance with the procedure of Example 8, step 1, methyl 3-oxobicyclo[2.2.1]heptane-2-carboxylate (446 mg, 2.65 mmol) was converted into 316 mg (40%) of methyl 3-(((trifluoromethyl)sulfonyl)oxy)bicyclo[2.2.1]hept-2-ene-2-carboxylate.

Step 3. Methyl 3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxylate



[0163] In accordance with the procedure of Example 8, step 2, methyl 3-(((trifluoromethyl)sulfonyl)oxy)bicyclo[2.2.1]hept-2-ene-2-carboxylate (160 mg, 0.533 mmol) and 3,5-dimethoxybenzylzinc(II) chloride (Aldrich, 1.3 mL of a 0.5 M solution in THF, 0.65 mmol) were converted after 3 days stirring at room temperature into 99 mg (61 %) of methyl 3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxylate after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 15% EtOAc/hexanes, gradient).

Step 4. 3-(3,5-Dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxylic acid



[0164] In accordance with the procedure of Example 4, step 2, methyl 3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxylate (98 mg, 0.32 mmol) was converted into 31.4 mg (34%) of 3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxylic acid and 60.5 mg (65%) of (E)-3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxylic acid (as a mixture of 2 stereoisomers) after purification on a Teledyne-Isco Combiflash machine (12 g gold column, hexanes → 35% EtOAc/hexanes, gradient).

Step 5. Methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxamido)methyl)benzoate



[0165] In accordance with the procedure of Example 3, step 2, 3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]-2-ene-2-carboxylic acid (31.4 mg, 0.109 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (22.8 mg, 0.11 mmol) were converted into 47 mg (quant.) of methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2. 1]hept-2-ene-2-carboxamido)methyl)benzoate.

Step 6. 4-((3-(3,5-Dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxamido)methyl)benzoic acid



[0166] In accordance with the procedure of Example 3, step 3, methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxamido)methyl)benzoate (9.2 mg, 0.021 mmol) was converted into 4.0 mg (45%) of the title compound.

Example 37


4-(((1S,2R,4R,E)-3-(3,5-Dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoic acid (racemic)



[0167] 


Step 1. (E)-Methyl 4-((3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate



[0168] In accordance with the procedure of Example 3, step 2, (E)-3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxylic acid (mixture of 2 stereoisomers, 60.5 mg, 0.210 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (43.6 mg, 0.210 mmol) were converted into 43.4 mg (48%) of racemic methyl 4-(((1S,2R,4R,E)-3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate and 23.3 mg (26%) of racemic methyl 4-(((1S,2S,4R,E)-3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate.

Step 2. 4-(((1S,2R,4R,E)-3-(3,5-Dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoic acid (racemic)



[0169] In accordance with the procedure of Example 3, step 3, racemic methyl 4-(((1S,2R,4R,E)-3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate (8.4 mg, 0.019 mmol) was converted into 3.1 mg (39%) of the title compound. Using the same conditions, racemic methyl 4-(((1S,2S,4R,E)-3-(3,5-dimethoxybenzylidene)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate (the other diastereomer from the previous step, 6.5 mg, 0.014 mmol) afforded 1.2 mg (19%) of the same title product.

Example 38


4-((3-(3,5-Dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0170] 


Step 1. Methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate



[0171] In accordance with the procedure of Example 5, step 1, methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]hept-2-ene-2-carboxamido)methyl)benzoate (40 mg, 0.092 mmol) was converted into 32 mg (80%) of methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate (high Rf diastereomer, presumably 1 racemic cis isomer) and 3.8 mg (10%) of methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate (low Rf diastereomer, mixture of 3 isomers, presumably the other racemic cis isomer and 2 trans isomers).

Step 2. 4-((2-(4-Methoxybenzyl)cyclohexanecarboxamido)methyl)benzoic acid (from high Rf ester diastereomer)



[0172] In accordance with the procedure of Example 3, step 3, methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate (high Rf diastereomer, 10.4 mg, 0.0238 mmol) was converted into 7.1 mg (70%) of the title compound.

Example 39


4-((3-(3,5-Dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoic acid (from low Rf ester diastereomer)



[0173] 



[0174] In accordance with the procedure of Example 3, step 3, methyl 4-((3-(3,5-dimethoxybenzyl)bicyclo[2.2.1]heptane-2-carboxamido)methyl)benzoate (low Rf diastereomer, mixture of 3 isomers, 3.8 mg, 0.0087 mmol) was converted into 1.4 mg (38%) of the title compound, still as a mixture of 3 isomers.

BIOLOGICAL DATA


Binding Data



[0175] Data from running binding and activity studies on the compounds of the invention were carried out as described in United States Patent No. 7,427,685.

Ki Binding Data



[0176] Competition binding experiments were performed in a medium containing Hank's balanced salt solution, Hepes 20 mM, pH 7.3, membranes (∼60 µg protein) or 2 × 105 cells from HEK 293 cells stably expressing human EP2 receptors, [3H]PGE2 (10 nM) and various concentrations of test compounds in a total volume of 300 µl. Reaction mixtures were incubated at 23 °C for 60 min, and were filtered over Whatman GF IB filters under vacuum. Filters were washed three times with 5 ml ice-cold buffer containing 50 mM Tris/HCl (PH 7.3). Non-specific binding was estimated in the presence of excess unlabeled PGE2 (10 µM). Binding data fitted to the binding model for a single class of binding sites, using nonlinear regression analysis. IC50 values thus obtained were converted to Ki using the equation of Ki=(IC50/(1+[L]/KD) where [L] represents PGE2 concentration (10 nM) and KD the dissociation constant for [3H]PGE2 at human EP2 receptors (40 nM).

Radioligand Binding


Cells Stably Expressing EP1, EP2, EP4, and FP Receptors



[0177] HEK-293 cells stably expressing the human or feline FP receptor, or EP1, EP2, or EP4 receptors were washed with TME buffer, scraped from the bottom of the flasks, and homogenized for 30 sec using a Brinkman PT 10/35 polytron. TME buffer was added to achieve a final 40 ml volume in the centrifuge tubes (the composition of TME is 100 mM TRIS base, 20 mM MgCl2, 2M EDTA; ION HCl is added to achieve a pH of 7.4).

[0178] The cell homogenate was centrifuged at 19000 rpm for 20 min at 4 °C using a Beckman Ti-60 rotor. The resultant pellet was resuspended in TME buffer to give a final 1 mg/ml protein concentration, as determined by Biorad assay. Radioligand binding competition assays vs. [3H-]17-phenyl PGF (5 nM) were performed in a 100µl volume for 60 min. Binding reactions were started by adding plasma membrane fraction. The reaction was terminated by the addition of 4 ml ice-cold TRIS-HCl buffer and rapid filtration through glass fiber GF/B filters using a Brandel cell harvester. The filters were washed 3 times with ice-cold buffer and oven dried for one hour.

[0179] [3H-] PGE2 (specific activity 180 Ci mmol) was used as the radio ligand for EP receptors. [3H] 17-phenyl PGF was employed for FP receptor binding studies. Binding studies employing EP1, EP2, EP4 and FP receptors were performed in duplicate in at least three separate experiments. A 200 µl assay volume was used. Incubations were for 60 min at 25 °C and were terminated by the addition of 4 ml of ice-cold 50 mM TRIS-HCl, followed by rapid filtration through Whatman GF/B filters and three additional 4 ml washes in a cell harvester (Brandel). Competition studies were performed using a final concentration of 5 nM [3H]-PGE2, or 5 nM [3H] 17-phenyl PGF and non-specific binding determined with 10-5 M of unlabeled PGE2, or 17-phenyl PGF, according to receptor subtype studied.

Methods for FLIPR™ Studies


(a) Cell Culture



[0180] HEK-293(EBNA) cells, stably expressing one type or subtype of recombinant human prostaglandin receptors (prostaglandin receptors expressed: hDP/Gqs5; hEP1; hEP2/Gqs5; hEP3A/Gqi5; hEP4/Gqs5; hFP; hIP; hTP), were cultured in 100 mm culture dishes in high-glucose DMEM medium containing 10% fetal bovine serum, 2 mM I-glutamine, 250 µg/ml geneticin (G418) and 200 µg/ml hygromycin B as selection markers, and 100 units/ml penicillin G, 100 µg/ml streptomycin and 0.25 µg/ml amphotericin B.

(b) Calcium Signal Studies on the FLIPR™



[0181] Cells were seeded at a density of 5 × 104 cells per well in Biocoat® Poly-D-lysine-coated black-wall, clear-bottom 96-well plates (Becton-Dickinson) and allowed to attach overnight in an incubator at 37 °C. Cells were then washed two times with HBSS-HEPES buffer (Hanks Balanced Salt Solution without bicarbonate and phenol red, 20 mM HEPES, pH 7.4) using a Denley Cellwash plate washer (Labsystems). After 45 minutes of dye-loading in the dark, using the calcium-sensitive dye Fluo-4 AM at a final concentration of 2 µM, plates were washed four times with HBSS-HEPES buffer to remove excess dye leaving 100 µL in each well. Plates were re-equilibrated to 37 °C for a few minutes.

[0182] Cells were excited with an Argon laser at 488 nm, and emission was measured through a 510-570 nm bandwidth emission filter (FLIPR™, Molecular Devices, Sunnyvale, CA). Drug solution was added in a 50 µL volume to each well to give the desired final concentration. The peak increase in fluorescence intensity was recorded for each well. On each plate, four wells each served as negative (HBSS-HEPES buffer) and positive controls (standard agonists: BW245C (hDP); PGE2(hEP1; hEP2/Gqs5; hEP3A/Gqi5; hEP4/Gqs5); PGF (hFP); carbacyclin (hIP); U-46619 (hTP), depending on receptor). The peak fluorescence change in each drug-containing well was then expressed relative to the controls.

[0183] Compounds were tested in a high-throughput (HTS) or concentration-response (CoRe) format. In the HTS format, forty-four compounds per plate were examined in duplicates at a concentration of 10-5 M. To generate concentration-response curves, four compounds per plate were tested in duplicates in a concentration range between 10-5 and 10-11. The duplicate values were averaged. In either, HTS or CoRe format each compound was tested on at least 3 separate plates using cells from different passages to give an n ≥ 3.

cAMP Assay



[0184] A 384-well drug plate was prepared to contain 6 test compounds, PGE2 and cAMP in 16 serial dilutions in triplicate, using a Biomek station. HEK-EBNA cells expressing a target PG receptor subtype (EP2 or EP4) were suspended in a stimulation buffer (HBSS, 0.1 % BSA, 0.5 mM IBMX and 5 mM HEPES, pH 7.4) in a density of 104 cells/5 µL. The reaction was initiated by mixing 5 µL drug dilutions with 5 µL ofHEK-EBNA cells in a well, carried out for 30 min at room temperature, and followed by the addition of 5 µL anti-cAMP acceptor beads in the control buffer with Tween-20 (25 mM NaCl, 0.03 % Tween-20, 5 mM HEPES, pH 7.4). After 30 min in the dark at room temperature, the mixtures were incubated with 15 µL biotinylated-cAMP/strepavidin donor beads in Lysis/Detection buffer (0.1 % BSA, 0.3 % Tween-20 and 5 mM HEPES, pH 7.4) for 45 min at the room temperature. Fluorescence changes were read using a Fusion-alpha HT microplate reader.

[0185] The results set forth below in Table 1 demonstrate that the compounds disclosed herein are selective prostaglandin EP4 agonists and antagonists, and are thus useful for the treatment of pathological conditions associated with EP4 receptors. In Table 1, "NA" indicates no activity.
Table 1
Ex.StructureEP2 DataEP4 dataOther Receptors (EC50 in nM)
cAMP EC50KicAMP EC50KIhFPhEP1hEP3AhTPhIPhDP
1

  6186 weak agonist 30 NA NA NA NA NA NA
2

  >10000 9 84 NA NA NA NA NA NA
3

33 1124 1 5 NA NA NA NA NA NA
4

2 109 >10000 7 NA NA NA NA NA NA
5

13 154 >10000 8 NA NA NA NA NA NA
6

10 733 >10000 18 NA NA NA NA >10000 NA
7

      1570            
8

2 177 0.2 1 NA NA NA NA NA NA
9

389 2148 >10000 2 NA NA NA NA NA NA
10

25 908 0.2 2 NA NA NA NA NA NA
11

48 2776 >10000 1 NA NA NA NA >10000 NA
12

1308 >10000 >10000 9 NA NA NA NA NA NA
13

573 6297 0.2 2 NA NA NA NA NA NA
14

  1194 >10000 45 NA NA NA NA NA NA
15

  7481 >10000 17 NA NA NA NA NA NA
16

  4119 >10000 114 NA NA NA NA NA NA
17

  403 >10000 8 NA NA NA NA NA NA
18

  462 >10000 16 NA NA NA NA NA NA
19

  393 0.2 2 NA NA NA NA NA NA
20

  >10000 >10000 0.2 NA NA NA NA NA NA
21

    >10000 4            
22

    >10000 4            
23

    >10000 8            
24

  83 >10000 4 NA NA NA NA NA NA
25

  4947 >10000 58 NA NA NA NA NA NA
26

  2923 35 116 NA NA NA NA NA NA
27

  138 >10000 2 NA NA NA NA NA NA
28

  2351 >10000 4 NA NA NA NA NA NA
29

  2574 4 19 NA NA NA NA NA NA
30

  219 >10000 6            
31

  994 >10000 5            
32

  1319 >10000 7            
33

  1142 >10000 1            
34

  >10000 >10000 44            
35

  >10000 >10000 86            
36

      1008            
37

  NA >10000 323            
38

  NA >10000 402            
39

  NA >10000 77            

Formulations and Compositions



[0186] Those skilled in the art will readily understand that for administration or the manufacture of medicaments the compounds disclosed herein can be admixed with pharmaceutically acceptable excipients which per se are known in the art. Specifically, a drug to be administered systemically, it may be confected as a powder, pill, tablet or the like, or as a solution, emulsion, suspension, aerosol, syrup or elixir suitable for oral or parenteral administration or inhalation.

[0187] For solid dosage forms or medicaments, non-toxic solid carriers include, but are not limited to, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, the polyalkylene glycols, talcum, cellulose, glucose, sucrose and magnesium carbonate. The solid dosage forms may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distcarate may be employed. They may also be coated by the technique described in the U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotic therapeutic tablets for control release. Liquid pharmaceutically administrable dosage forms can, for example, comprise a solution or suspension of one or more of the presently useful compounds and optional pharmaceutical adjutants in a carrier, such as for example, water, saline, aqueous dextrose, glycerol, ethanol and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like. Typical examples of such auxiliary agents are sodium acetate, sorbitan monolaurate, triethanolamine, sodium acetate, triethanolamine oleate, etc. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 16th Edition, 1980. The composition of the formulation to be administered, in any event, contains a quantity of one or more of the presently useful compounds in an amount effective to provide the desired therapeutic effect.

[0188] Parenteral administration is generally characterized by injection, either subcutaneously, intramuscularly or intravenously. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol and the like. In addition, if desired, the injectable pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like.

[0189] The amount of the compound or compounds administered is, of course, dependent on the therapeutic effect or effects desired, on the specific mammal being treated, on the severity and nature of the mammal's condition, on the manner of administration, on the potency and pharmacodynamics of the particular compound or compounds employed, and on the judgment of the prescribing physician. The therapeutically effective dosage of the compounds can be in the range of about 0.5 or about 1 to about 100 mg/kg/day.

[0190] In some embodiments, the amount of the active compound in a pharmaceutical composition is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 3.0%, 4.0% and 5.0% w/w.

[0191] In some embodiments, an effective amount, e.g., a therapeutically effective amount, of the active compound in a pharmaceutical composition is afforded at a concentration of about 1x10-7 to 50% (w/w), about 0.001 to 50% (w/w), about 0.01 to 50% (w/w), about 0.1 to 50% (w/w), or about 1 to 50% (w/w). In some embodiments, the therapeutically effective amount of the active compound in a pharmaceutical composition is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% and 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 3.0%, 4.0% and 5.0% w/w.

[0192] A liquid which is ophthalmically acceptable is formulated such that it can be administered topically to the eye. The comfort should be maximized as much as possible, although sometimes formulation considerations (e.g. drug stability) may necessitate less than optimal comfort. In the case that comfort cannot be maximized, the liquid should be formulated such that the liquid is tolerable to the patient for topical ophthalmic use. Additionally, an ophthalmically acceptable liquid should either be packaged for single use, or contain a preservative to prevent contamination over multiple uses.

[0193] For ophthalmic application, solutions or medicaments are often prepared using a physiological saline solution as a major vehicle. Ophthalmic solutions can be maintained at a comfortable pH with an appropriate buffer system. The formulations may also contain conventional, pharmaceutically acceptable preservatives, stabilizers and surfactants.

[0194] Preservatives that may be used in the pharmaceutical compositions of the present invention include, but are not limited to, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetate and phenylmercuric nitrate. A useful surfactant is, for example, Tween 80. Likewise, various useful vehicles may be used in the ophthalmic preparations of the present invention. These vehicles include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropyl methylcellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl cellulose and purified water.

[0195] Tonicity adjustors may be added as needed or convenient. They include, but are not limited to, salts, particularly sodium chloride, potassium chloride, mannitol and glycerin, or any other suitable ophthalmically acceptable tonicity adjustor.

[0196] Various buffers and means for adjusting pH may be used so long as the resulting preparation is ophthalmically acceptable. Accordingly, buffers include acetate buffers, citrate buffers, phosphate buffers and borate buffers. Acids or bases may be used to adjust the pH of these formulations as needed.

[0197] In a similar vein, an ophthalmically acceptable antioxidant for use in the present invention includes, but is not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene.

[0198] Other excipient components which may be included in the ophthalmic preparations are chelating agents. A useful chelating agent is edetate disodium, although other chelating agents may also be used in place or in conjunction with it.

[0199] The ingredients are usually used in the following amounts:
IngredientAmount (% w/v)
active ingredient about 0.001-5
preservative 0-0.10
vehicle 0-40
tonicity adjustor 1-10
buffer 0.01-10
pH adjustor q.s. pH 4.5-7.5
antioxidant as needed
surfactant as needed
purified water as needed to make 100%


[0200] For topical use, creams, ointments, gels, solutions or suspensions, etc., containing the compound disclosed herein are employed. Topical formulations may generally be comprised of a pharmaceutical carrier, cosolvent, emulsifier, penetration enhancer, preservative system, and emollient.

[0201] The compositions may be administered between 1 and 7 days a week, for a period of time necessary to achieve the desired results, which may be several days to several months. The compositions can be administered once or several times (2, 3, 4, or more times) a day depending on the desired effect. In certain embodiments, the compositions can be administered every 1, 2, 3, 4, 5, 6, or 7 days. In another embodiment, the compositions can be administered one or more times every 1, 2, 3, or 4 weeks. The administration can be on a monthly or bi-monthly basis. Further, the compositions can be administered for 1, 2, 3, 6, 9, or 12 months or more. In certain embodiments, the compositions can be administered on an ongoing basis to maintain a desired result.

[0202] The disclosed compounds can be administered as part of a composition. As used herein, "formulation" and "composition" may be used interchangeably and refer to a combination of elements that is presented together for a given purpose. Such terms are well known to those of ordinary skill in the art.

[0203] As used herein, "carrier," "inert carrier," and "acceptable carrier" may be used interchangeably and refer to a carrier which may be combined with the presently disclosed compounds in order to provide a desired composition. Those of ordinary skill in the art will recognize a number of carriers that are well known for making specific pharmaceutical and/or cosmetic compositions. Desirably, the carrier is suitable for application to keratinous surfaces or other areas of the body. Upon application, acceptable carriers are substantially free of adverse reactions with skin and other keratinous surfaces. For example, the carriers may take the form of fatty or non-fatty creams, milky suspensions or emulsion-in-oil or oil-in-water types, lotions, gels or jellies, colloidal or non-colloidal aqueous or oily solutions, pastes, aerosols, soluble tablets or sticks. In accordance with one embodiment, the composition includes a dermatologically compatible vehicle or carrier. The vehicle which may be employed for preparing compositions may comprise, for example, aqueous solutions such as e.g., physiological salines, oil solutions or ointments. The vehicle furthermore may contain dermatologically compatible preservatives such as e.g., benzalkonium chloride, surfactants like e.g., polysorbate 80, liposomes or polymers, for example, methyl cellulose, polyvinyl alcohol, polyvinyl pyrrolidone and hyaluronic acid; these may be used for increasing the viscosity.

[0204] Examples of additional agents which can be included in the present compositions are anti-itch, anti-cellulite, anti-scarring, and anti-inflammatory agents, anesthetics, anti-irritants, vasoconstrictors, vasodilators, as well as agents to prevent/stop bleeding, and improve/remove pigmentation, moisturizers, desquamating agents, tensioning agents, anti-acne agents. Anti-itch agents can include methyl sulphonyl methane, sodium bicarbonate, calamine, allantoin, kaolin, peppermint, tea tree oil and combinations thereof. Anti-cellulite agents can include forskolin, xanthine compounds such as, but not limited to, caffeine, theophylline, theobromine, and aminophylline, and combinations thereof. Anesthetic agents can include lidocaine, benzocaine, butamben, dibucaine, oxybuprocaine, pramoxine, proparacaine, proxymetacaine, tetracaine, and combinations thereof. Anti-scarring agents can include IFN-.gamma., fluorouracil, poly(lactic-co-glycolic acid), methylated polyethylene glycol, polylactic acid, polyethylene glycol and combinations thereof. Anti-inflammatory agents can include dexamethasone, prednisolone, corticosterone, budesonide, estrogen, sulfasalazine, mesalamine and derivatives and combinations thereof. Additionally, active agents such as epinephrine, thymidine, cytidine, uridine, antiypyrin, aminocaproic acid, tranexamic acid, eucalyptol, allantoin, glycerin, and sodium selenite, can be included. Formulations can further comprise degradation inhibitors. Degradation inhibitors, include but are not limited to, glycosaminoglycans (e.g., heparin, heparin sulfate, dermatan sulfate, chrondroitin sulfate, o-sulfated HA, lnamarin, and amygdalin), antioxidants (e.g. ascorbic acid, melatonin, vitamin C, vitamin E), proteins (e.g., serum hyaluronidase inhibitor), and fatty acids (e.g. saturated C10 to C22 fatty acids). In certain embodiments, additional active agent is an antioxidant. In certain embodiments, the antioxidant comprises a vitamin C and/or a vitamin E such as d-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS).

[0205] The disclosed compositions are well suited for topical, subcutaneous, intradermal, subdermal, subcutaneous, and transdermal administration. Topical administration relates to the use of a composition applied to the surface of the skin at the site of a skin blemish for exertion of local action. Accordingly, such topical compositions include those pharmaceutical or cosmetic forms in which the composition is applied externally by direct contact with the skin surface to be treated, such as the face, neck, arms, legs, and/or torso. Conventional pharmaceutical or cosmetic forms for this purpose include ointments, liniments, creams, shampoos, lotions, pastes, jellies, sprays, aerosols, and the like, and may further be applied directly or in patches or impregnated dressings depending on blemish and skin region to be treated. The term "ointment" embraces formulations (including creams) having oleaginous, water-soluble and emulsion-type bases, e.g., petrolatum, lanolin, polyethylene glycols, as well as mixtures of these.

[0206] The compositions are appropriate for mesotherapy applications as well. Mesotherapy is a non-surgical cosmetic treatment technique involving intra-epidermal, intra-dermal, and/or subcutaneous injection of a composition. The compositions are administered in the form of small multiple droplets into the epidermis, dermo-epidermal junction, and/or the dermis.

[0207] The actual dose of the active compounds of the present invention depends on the specific compound, and on the condition to be treated; the selection of the appropriate dose is well within the knowledge of the skilled artisan.

[0208] The compounds disclosed herein are also useful in combination with other drugs useful for the treatment of glaucoma or other conditions.

[0209] For the treatment of glaucoma, combination treatment with the following classes of drugs are contemplated:

β-Blockers (or β-adrenergic antagonists) including carteolol, levobunolol, metiparanolol, timolol hemihydrate, timolol maleate, β1-selective antagonists such as betaxolol, and the like, or pharmaceutically acceptable salts or prodrugs thereof;

Adrenergic Agonists including non-selective adrenergic agonists such as epinephrine borate, epinephrine hydrochloride, and dipivefrin, and the like, or pharmaceutically acceptable salts or prodrugs thereof; and

α2-selective adrenergic agonists such as apraclonidine, brimonidine, and the like, or pharmaceutically acceptable salts or prodrugs thereof;

Carbonic Anhydrase Inhibitors including acetazolamide, dichlorphenamide, methazolamide, brinzolamide, dorzolamide, and the like, or pharmaceutically acceptable salts or prodrugs thereof;

Cholinergic Agonists including direct acting cholinergic agonists such as carbachol, pilocarpine hydrochloride, pilocarbine nitrate, pilocarpine, and the like, or pharmaceutically acceptable salts or prodrugs thereof;

chlolinesterase inhibitors such as demecarium, echothiophate, physostigmine, and the like, or pharmaceutically acceptable salts or prodrugs thereof;

Glutamate Antagonists and other neuroprotective agents such as Ca2+ channel blockers such as memantine, amantadine, rimantadine, nitroglycerin, dextrophan, dextromethorphan, CGS-19755, dihydropyridines, verapamil, emopamil, benzothiazepines, bepridil, diphenylbutylpiperidines, diphenylpiperazines, HOE 166 and related drugs, fluspirilene, eliprodil, ifenprodil, CP-101,606, tibalosine, 2309BT, and 840S, flunarizine, nicardipine, nifedimpine, nimodipine, barnidipine, verapamil, lidoflazine, prenylamine lactate, amiloride, and the like, or pharmaceutically acceptable salts or prodrugs thereof;

Prostamides such as bimatoprost, or pharmaceutically acceptable salts or prodrugs thereof; and

Prostaglandins including travoprost, UFO-21, chloprostenol, fluprostenol, 13,14-dihydro-chloprostenol, isopropyl unoprostone, latanoprost and the like.

Cannabinoids including CB1 agonists such as WIN-55212-2 and CP-55940 and the like, or pharmaceutically acceptable salts or prodrugs thereof.



[0210] For treatment of diseases affecting the eye including glaucoma, these compounds can be administered topically, periocularly, intraocularly, or by any other effective means known in the art.

[0211] Pathological conditions associated with EP4 receptors include, but are not limited to, acute hepatitis, asthma, bronchitis, burn, chronic obstructive respiratory diseases, Crohn's disease, digestive ulcer, glaucoma (and other diseases related to elevated intraocular pressure), hemophagous syndrome, hepatopathy, hypercytokinemia at dialysis, hypertension, immunological diseases (autoimmune diseases, organ transplantation, etc.), inflammation (such as rheumatoid arthritis), Kawasaki disease, liver injury, macrophage activation syndrome, myocardial ischemia, nephritis, nerve cell death, osteoporosis and diseases associated with bone disorders, premature birth, pulmonary emphysema, pulmonary fibrosis, pulmonary injury, renal failure, sepsis, sexual dysfunction, shock, sleep disorder, Still disease, stomatitis , systemic granuloma, systemic inflammatory syndrome, thrombosis and stroke, and ulcerative colitis.

[0212] The foregoing descriptions details specific methods and compositions that can be employed to practice the present invention, and represents the best mode contemplated. It should not be construed as limiting the overall scope hereof; rather, the ambit of the present invention is to be governed only by the lawful construction of the appended claims.


Claims

1. A compound having the following structure:

or a pharmaceutically acceptable salt or diastereomer or enantiomer thereof, wherein:

the dashed line is a single or a double bond;

Y is H or CH3;

X is zero, one or two substituents on the cycloalkane or cycloalkene ring, said substituents chosen from the group consisting of H, C1 - C6 alkyl, hydroxyalkyl, aryl, halogen, cycloalkene, CF3, C(O)R, COCF3, SO2N(R)2, SO2NH2, NO2, and CN;

n is selected from the group consisting of 0, 1, 2, 3, or 4;

R is C1 - C6 alkyl;

R2 is selected from the group consisting of H, C1 - C6 alkyl, hydroxyalkyl, phenyl or biphenyl;

R3 is selected form the group consisting of H, C(O)R5, SO2R5, C1 - C6 alkyl;

R5 is C1-C6 alkyl, haloalkyl including trifluoromethyl, aryl or heteroaryl;

R6 is CO2H, CH3, CO2R2, CH2OR2, CONR2R3 or tetrazol-5-yl;

R7 is selected from the group consisting of H, CF3, OCH3; and

R8 is selected form the group consisting of H and OCH3.


 
2. The compound of claim 1, wherein R6 is CO2H and n is selected from the group consisting of 0, 1 and 2.
 
3. The compound of claim 1, wherein n is selected from the group consisting of 0, 1 and 2, preferably 1 and 2, and the dashed line represents a double bond.
 
4. The compound of claim 3, wherein R7 and R8 are OCH3 or wherein R7 is CF3 and R8 is H.
 
5. The compound of claim 1, wherein (i) the dashed line represents a single bond; or wherein (ii) n is selected from the group consisting of 3 and 4; or wherein (iii) R7 and R8 are OCH3.
 
6. The compound of claim 1 selected from the group consisting of:

























and pharmaceutically acceptable salts, diastereomers, and enantiomers thereof.
 
7. A pharmaceutical composition comprising a compound of any one of claims 1-6 and a pharmaceutically acceptable composition.
 
8. The compound of any one of claims 1-6 or the pharmaceutical composition of claim 7 for use in a method of treating a disease.
 
9. The compound of any one of claims 1-6 or the pharmaceutical composition of claim 7 for use in a method of treating glaucoma, the method comprising administering an effective amount of said compound or said pharmaceutical composition to an individual in need thereof.
 


Ansprüche

1. Verbindung mit der nachstehenden Struktur:

oder ein pharmazeutisch annehmbares Salz oder Diastereomer oder Enantiomer davon, worin:

die gestrichelte Linie eine Einfachbindung oder eine Doppelbindung ist;

Y H oder CH3 ist;

X null, einer oder zwei Substituenten an dem Cycloalkan- oder Cycloalkenring ist, wobei der/die Substituent(en) aus der Gruppe bestehend aus H, C1-6-Alkyl, Hydroxyalkyl, Aryl, Halogen, Cycloalken, CF3, C(O)R, COCF3, SO2N(R)2, SO2NH2, NO2 und CN ausgewählt ist;

n aus der Gruppe bestehend aus 0, 1, 2, 3 oder 4 ausgewählt ist;

R C1-6-Alkyl ist;

R2 aus der Gruppe bestehend aus H, C1-6-Alkyl, Hydroxyalkyl, Phenyl oder Biphenyl ausgewählt ist;

R3 aus der Gruppe bestehend aus H, C(O)R5, SO2R5, C1-6-Alkyl ausgewählt ist;

R5 C1-6-Alkyl, Halogenalkyl, einschliesslich Trifluormethyl, Aryl oder Heteroaryl, ist;

R6 CO2H, CH3, CO2R2, CH2OR2, CONR2R3 oder Tetrazol-5-yl ist;

R7 aus der Gruppe bestehend aus H, CF3 und OCH3 ausgewählt ist; und

R8 aus der Gruppe bestehend aus H und OCH3 ausgewählt ist.


 
2. Verbindung gemäss Anspruch 1, worin R6 CO2H ist und n aus der Gruppe bestehend aus 0, 1 und 2 ausgewählt ist.
 
3. Verbindung gemäss Anspruch 1, worin n aus der Gruppe bestehend aus 0, 1 und 2, vorzugsweise 1 und 2, ausgewählt ist und die gestrichelte Linie eine Doppelbindung darstellt.
 
4. Verbindung gemäss Anspruch 3, worin R7 und R8 OCH3 sind oder worin R7 CF3 ist und R8 H ist.
 
5. Verbindung gemäss Anspruch 1, worin (i) die gestrichelte Linie eine Einfachbindung darstellt oder worin (ii) n aus der Gruppe bestehend aus 3 und 4 ausgewählt ist oder worin (iii) R7 und R8 OCH3 sind.
 
6. Verbindung gemäss Anspruch 1, ausgewählt aus der Gruppe bestehend aus:

























und pharmazeutisch annehmbare Salze, Diastereomere und Enantiomere davon.
 
7. Pharmazeutische Zusammensetzung, umfassend eine Verbindung gemäss irgendeinem der Ansprüche 1 bis 6 und eine pharmazeutisch annehmbare Zusammensetzung.
 
8. Verbindung gemäss irgendeinem der Ansprüche 1 bis 6 oder pharmazeutische Zusammensetzung gemäss Anspruch 7 zur Verwendung in einem Verfahren zur Behandlung einer Erkrankung.
 
9. Verbindung gemäss irgendeinem der Ansprüche 1 bis 6 oder pharmazeutische Zusammensetzung gemäss Anspruch 7 zur Verwendung in einem Verfahren zur Behandlung von Glaukom, wobei das Verfahren die Verabreichung einer wirksamen Menge der Verbindung oder der pharmazeutischen Zusammensetzung an ein Individuum, das dies benötigt, umfasst.
 


Revendications

1. Composé ayant la structure suivante :

ou un sel pharmaceutiquement acceptable ou diastéréoisomère ou énantiomère de celui-ci, dans lequel :

la ligne pointillée est une liaison double ou simple ;

Y est H ou CH3 ;

X est zéro, un ou deux substituants sur le cycle cycloalcane ou cycloalcène, lesdits substituants choisis parmi le groupe constitué de H, C1-C6 alkyle, hydroxyalkyle, aryle, halogène, cycloalcène, CF3, C(O)R, COCF3, SO2N(R)2, SO2NH2, NO2, et CN ;

n est choisi parmi le groupe constitué de 0, 1, 2, 3, ou 4 ;

R est C1-C6 alkyle ;

R2 est choisi parmi le groupe constitué de H, C1-C6 alkyle, hydroxyalkyle, phényle ou biphényle ;

R3 est choisi parmi le groupe constitué de H, C(O)R5, SO2R5, C1-C6 alkyle ;

R5 est C1-C6 alkyle, haloalkyle comportant trifluorométhyle, aryle ou hétéroaryle ;

R6 est CO2H, CH3, CO2R2, CONR2R3 ou tétrazol-5-yle ;

R7 est choisi parmi le groupe constitué de H, CF3, OCH3 ; et

R8 est choisi parmi le groupe constitué de H et OCH3.


 
2. Le composé de la revendication 1, dans lequel R6 est CO2H et n est choisi parmi le groupe constitué de 0, 1 et 2.
 
3. Le composé de la revendication 1, dans lequel n est choisi parmi le groupe constitué de 0, 1 et 2, préférablement 1 et 2, et les lignes pointillés représentent une liaison double.
 
4. Le composé de la revendication 3, dans lequel R7 et R8 sont OCH3 ou dans lequel R7 est CF3 et R8 est H.
 
5. Le composé de la revendication 1, dans lequel (i) la ligne pointillée représente une liaison simple ; ou dans lequel (ii) n est choisi parmi le groupe constitué de 3 et 4 ; ou dans lequel (iii) R7 et R8 sont OCH3.
 
6. Le composé de la revendication 1 choisi parmi le groupe constitué de :

























et des sels pharmaceutiquement acceptables, diastéréoisomères, et énantiomères de celui-ci.
 
7. Composition pharmaceutique comprenant un composé de l'une quelconque des revendications 1-6 et une composition pharmaceutiquement acceptable.
 
8. Le composé de l'une quelconque des revendications 1-6 ou la composition pharmaceutique de la revendication 7pour l'utilisation dans une méthode de traitement d'une maladie.
 
9. Le composé de l'une quelconque des revendications 1-6 ou la composition pharmaceutique de la revendication 7 pour l'utilisation dans une méthode de traitement du glaucome, la méthode comprenant l'administration d'une quantité efficace dudit composé ou de ladite composition pharmaceutique à un individu qui en a besoin.
 






Cited references

REFERENCES CITED IN THE DESCRIPTION



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Patent documents cited in the description




Non-patent literature cited in the description