4-Amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one derivatives and their salts

Abstract

The present disclosure relates to variants or derivatives of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one, and salts thereof, for use as agonists and antagonists. The present disclosure further relates to compositions, methods of manufacture, and methods of treatment.

Description

[Technical field]

[0001] This application claims priority to U.S. Provisional Application No. 63 / 130,010, filed December 23, 2020, and U.S. Provisional Application No. 63 / 245,288, filed September 17, 2021.

[0002] The present disclosure relates to variants or derivatives of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one, and salts thereof, for use as agonists and antagonists. The present disclosure further relates to pharmaceutical compositions, methods of manufacture, and methods of treatment. [Background technology]

[0003] Small molecule drugs have proven useful for a variety of diseases and conditions. Some act as antagonists by blocking or attenuating a biological response, some act as agonists by activating a biological response, and some can function as both.

[0004] While many small molecule drugs are known, new and diverse therapeutic agents are needed. For example, according to the National Center for Complementary and Integrative Health, multiple sclerosis ("MS") affects approximately 2,500,000 people worldwide, including approximately 400,000 in the United States. Although there are several drugs available for the treatment of MS, not everyone responds well to the available drug therapies. Examples of other diseases and conditions for which there is a need include autoimmune disorders such as lupus, pemphigus vulgaris, myasthenia gravis, Sjogren's syndrome, dry eye, multiple sclerosis, Wegener's granulomatosis, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, granulomatosis with polyangiitis, and rheumatoid arthritis.

[0005] The present disclosure provides compounds that are tyrosine kinase inhibitors, particularly Bruton's tyrosine kinase ("BTK") inhibitors, and thus are useful in the treatment of diseases such as cancer, autoimmune diseases, inflammatory diseases, and thromboembolic diseases. Also provided are pharmaceutical compositions containing such compounds and methods for producing such compounds.

[0006] BTK is a member of the Tec family of non-receptor tyrosine kinases and is essential for downstream B cell signaling from the B cell receptor. BTK is expressed in B cells and other hematopoietic cells such as monocytes, macrophages, and mast cells. BTK functions in various aspects of B cell function, maintaining the B cell repertoire (see Non-Patent Document 1). B cells play a role in other autoimmune diseases such as rheumatoid arthritis (see Non-Patent Document 2, Non-Patent Document 3, and Non-Patent Document 4), systemic lupus erythematosus, and cancer (see Non-Patent Document 5, Non-Patent Document 6, and Non-Patent Document 7).

[0007] BTK inhibitors may also have potential for the treatment of allergic diseases (Non-Patent Document 4). It has been noted that irreversible inhibitors suppress passive cutaneous anaphylaxis (PCA) induced by IgE-antigen complexes in mice. These findings are consistent with those observed in BTK mutant mast cells and knockout mice, suggesting that BTK inhibitors may be useful in the treatment of asthma, an IgE-dependent allergic disease of the airways.

[0008] Therefore, compounds that inhibit BTK are believed to be useful in treating diseases such as autoimmune diseases, inflammatory diseases, and cancer.

[0009] It has been surprisingly discovered that certain 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one derivatives function as agonists, antagonists, or both, of multiple receptors involved in many diseases and disorders, including those listed above. [Prior art documents] [Non-patent literature]

[0010] [Non-Patent Document 1] Gauld SB et al. B cell antigen receptor signaling: roles in cell development and disease.Science, 296:1641~2.2002 [Non-Patent Document 2] Perosa F. et al., CD20-depleting therapy in autoimmune diseases:from basic research to the clinic.J Intern Med.267:260~77.2010 [Non-Patent Document 3] Dorner T et al. Targeting B cells in immune-mediated inflammatory disease: a comprehensive review of mechanisms of action and identification of biomarkers.Pharmacol Ther.125:464~75.2010 [Non-Patent Document 4] Honigberg, L. et al., The selective BTK inhibitor PCI-32765 blocks B cell and mast cell activation and prevents mouse collagen-induced arthritis.Clin.Immunol.127 S1:S111.2008 [Non-Patent Document 5] Shlomchik MJ et al. The role of B cells in lpr / lpr-induced autoimmunity.J.Exp Med.180:1295~1306.1994 [Non-Patent Document 6] Honigberg LA, The Bruton tyrosine kinase inhibitor PCI-32765 blocks B-cell activation and is efficacious in models of autoimmune disease and B-cell malignancy.Proc.Natl.Acad.Sci.107:13075~80.2010 [Non-Patent Document 7] Mina-Osorio P et al., Suppression of glomerulonephritis in lupus-prone NZB x NZW mice by RN486, a selective inhibitor of Bruton's tyrosine kinase.Arthritis Rheum.65:2380~91.2013 Summary of the Invention [Means for solving the problem]

[0011] Described herein, in certain embodiments, are derivatives of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one and salts thereof.

[0012] In one embodiment, the compound of formula I is provided herein: [ka] or a salt thereof is disclosed.

[0013] In one embodiment, the compound of formula Ia: [ka] or a salt thereof. In another embodiment, the compound of formula Ia or a salt thereof is about 90% pure.

[0014] In another embodiment, the compound of formula Ib is provided herein: [ka] or a salt thereof, wherein the compound or salt thereof is about 90% pure.

[0015] In a further embodiment, the compound of formula II is provided herein. [ka] or a salt thereof, wherein the compound or salt thereof is about 99.6% pure.

[0016] In another embodiment, the composition described herein comprises a medicament comprising a pharma- ceutically acceptable excipient and [ka] and at least one compound selected from, or a salt thereof.

[0017] In a further embodiment, provided herein is a method for preparing a compound of formula Ia or a salt thereof, comprising the steps of: [ka] A method is disclosed that includes reacting a compound of the formula: with acryloyl chloride.

[0018] In a further embodiment, provided herein is a method for preparing a compound of formula Ib or a salt thereof, comprising the steps of: [ka] A method is disclosed that includes reacting a compound of the formula: with acryloyl chloride.

[0019] Additional objects and advantages will be set forth in part in the description which follows, and in part will be understood from the description, or may be learned by practice. The objects and advantages will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.

[0020] 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 scope of the claims. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] I. Definition Unless otherwise stated, the following terms used in the specification and claims are defined for purposes of this disclosure and have the following meanings:

[0022] A "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" means a carrier or excipient that is generally safe, non-toxic, and not biologically or otherwise undesirable and is useful in preparing pharmaceutical compositions, and includes carriers or excipients that are acceptable for veterinary as well as human pharmaceutical use. As used in the specification and claims, a "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes both one and more of such carriers or excipients.

[0023] "Treating" a disease, or its "treatment," includes the following: (1) To prevent disease, e.g., to prevent the development of clinical symptoms of disease in a patient, e.g., a human patient, who may have been exposed to or be predisposed to the disease but has not yet experienced or exhibited symptoms of the disease; (2) inhibiting the disease, e.g., preventing or reducing the onset of the disease or its clinical symptoms; or (3) Alleviating the disease, e.g., causing regression of the disease or its clinical symptoms.

[0024] "Optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances in which the event or circumstance occurs and instances in which the event or circumstance does not occur.

[0025] "Therapeutically effective amount" means an amount of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one derivative or salt thereof that, when administered to a patient, e.g., a human patient, for treating a disease, is sufficient to affect such treatment of the disease. A "therapeutically effective amount" varies depending on the compound, the disease and its severity, and the age, weight, etc., of the patient, e.g., a human patient, being treated.

[0026] "About X% purity" or "about X% purity" (where "X" is a number) means that the reference compound is present at about X% as determined by HPLC area of ​​total peak area (detected by UV at 254 nm).

[0027] Before describing the present teachings in detail, it should be understood that the present disclosure is not limited to specific compositions or method steps, as such may vary. As used in this specification and the appended claims, it should be noted that the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to a "conjugate" includes a plurality of conjugates, reference to a "cell" includes a plurality of cells, and so forth.

[0028] Numerical ranges are inclusive of the numbers defining the range. Measurements and measurables are understood to be approximations taking into account significant digits and error associated with measurement. Furthermore, the use of "comprise", "comprises", "comprising", "contain", "contains", "containing", "include", "includes" and "including" are not intended to be limiting. It is understood that both the foregoing general and detailed description are exemplary and explanatory only and are not intended to be restrictive of the teachings.

[0029] Unless otherwise indicated, embodiments described herein as "comprising" various components are also intended to be "consisting of" or "consisting essentially of" the listed components; embodiments described herein as "consisting of" various components are also intended to be "comprising" or "consisting essentially of" the listed components; and embodiments described herein as "consisting essentially of" various components are also intended to be "consisting of" or "comprising" the listed components (this interchangeability does not apply to the use of these terms in the claims).

[0030] "Or" is used in its inclusive sense, ie, equivalent to "and / or", unless the context requires otherwise.

[0031] II. Compounds In one aspect, provided herein are compounds that are 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one derivatives and salts thereof. In some embodiments, the compounds are about 50% pure. In some embodiments, the compounds are about 55% pure. In some embodiments, the compounds are about 60% pure. In some embodiments, the compounds are about 65% pure. In some embodiments, the compounds are about 70% pure. In some embodiments, the compounds are about 75% pure. In some embodiments, the compounds are about 80% pure. In some embodiments, the compounds are about 85% pure. In some embodiments, the compounds are about 90% pure. In some embodiments, the compounds are about 95% pure. In some embodiments, the compounds are about 98% pure. In some embodiments, the compounds are about 99% pure.

[0032] In one aspect, the present invention provides a compound of formula I: [ka] or a salt thereof.

[0033] In one aspect, the present disclosure provides a compound of formula Ia: [ka] or a salt thereof. In some embodiments, the compound of formula Ia or a salt thereof is about 90% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 95% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 98% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 99% pure.

[0034] In one aspect, the present disclosure provides a compound of formula Ib: [ka] or a salt thereof, wherein the compound or salt thereof is about 90% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 95% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 98% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 99% pure.

[0035] In one aspect, the present invention relates to a compound represented by formula II [ka] or a salt thereof, wherein the compound or salt thereof is about 99.6% pure. In some embodiments, the compound is [ka] or a salt thereof, wherein the compound or salt thereof is about 90% pure. In some embodiments, the compound of formula IIa or a salt thereof is about 95% pure. In some embodiments, the compound of formula IIa or a salt thereof is about 98% pure. In some examples, the compound is [ka] or a salt thereof, wherein the compound or salt thereof is about 90% pure. In some embodiments, the compound of formula IIb or a salt thereof is about 95% pure. In some embodiments, the compound of formula IIb or a salt thereof is about 98% pure.

[0036] In one aspect, the present disclosure provides compounds of formula III, formula IV, and formula V [ka] or a salt thereof.

[0037] III. Composition In one aspect, provided herein is a pharmaceutical composition comprising a pharma- ceutically acceptable excipient and a compound or salt thereof that is a variant or derivative of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one, which the inventors have discovered as an in vivo metabolite of (R)-1-(1-acryloylpiperidin-3-yl)-4-amino-3-(4-phenoxyphenyl)-1H-imidazo[4,5-c]pyridin-2(3H)-one.

[0038] In one embodiment, a pharmaceutical composition as described herein comprises a pharmacopoeitic agent comprising: [ka] and at least one compound selected from, or a salt thereof. In some embodiments, the at least one compound, or a salt thereof, is present in an effective amount.

[0039] In one embodiment, a pharmaceutical composition as described herein comprises a pharmacopoeitic agent comprising: [ka] and at least one compound selected from, or a salt thereof.

[0040] In some embodiments, at least one compound or its salt is present in an effective amount.In one embodiment, a composition is provided that includes a pharma-ceutically acceptable excipient and a compound of formula Ia or its salt.In some embodiments, the compound of formula Ia or its salt is at least about 98% pure.

[0041] In one embodiment, provided herein is a composition comprising a pharma- ceutically acceptable excipient and a compound of formula Ib, or a salt thereof. In some embodiments, the compound of formula Ib, or a salt thereof, is at least about 99% pure.

[0042] Pharmaceutical compositions according to the present disclosure may be in a form suitable for oral, buccal, parenteral, nasal, topical, or rectal administration, or in a form suitable for administration by inhalation.

[0043] The compounds described herein can be used to manufacture compositions, such as pharmaceutical compositions, by combining a compound as an active ingredient with a pharma- ceutically acceptable excipient. Some examples of materials which may function as pharma- ceutically acceptable excipients include: sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; surfactants such as polysorbate 80 (i.e., Tween 80); powdered tragacanth; malt; gelatin; talc; suppository bases such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffers; polyesters, polycarbonates or polyanhydrides; and other non-toxic compatible substances used in pharmaceutical compositions. Pharmaceutical compositions can be prepared by known pharmaceutical methods. Suitable formulations are described, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st Edition (2005), which is incorporated herein by reference.

[0044] Wetting agents, emulsifying agents, and lubricating agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives, and antioxidants can also be present in the composition.

[0045] Examples of pharma- ceutically acceptable antioxidants include: water soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; oil soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha tocopherol, etc.; and metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0046] Compositions of the present disclosure suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an emulsion in an oil-in-water or water-in-oil liquid, or as an elixir or syrup, or as a pastille (using an inert base such as gelatin and glycerin, or sucrose and acacia), or as a mouthwash, etc., each containing a predetermined amount of a compound of the present disclosure as an active ingredient. Compounds of the present disclosure may also be administered as a bolus, electuary, or paste.

[0047] In solid dosage forms of the present disclosure for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharma- ceutically acceptable carriers such as sodium citrate or dicalcium phosphate, or any of the following: fillers or extenders, for example starches, lactose, sucrose, glucose, mannitol, or silicic acid; binders, for example carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, or acacia; humectants, for example glycerin; disintegrating agents, for example agar-agar, calcium carbonate, potato starch, or tapioca starch, alginic acid, certain silicates, and sodium carbonate; Solution retarding agents such as paraffin; absorption enhancers such as quaternary ammonium compounds; wetting agents such as cetyl alcohol, glycerol monostearate, and nonionic surfactants; absorbents such as kaolin and bentonite clay; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and coloring agents. In the case of capsules, tablets, and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type may also be used as fillers for soft and hard shell gelatin capsules using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols.

[0048] Tablets can be produced by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be produced using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents or dispersing agents. Molded tablets can be produced in a suitable machine by moistening a mixture of powdered compounds with an inert liquid diluent.

[0049] Tablets and other solid dosage forms of the pharmaceutical composition of the present disclosure, such as dragees, capsules, pills and granules, may be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may also be formulated for sustained or controlled release of the active ingredient therein, for example, using hydroxypropylmethylcellulose in varying proportions to obtain the desired release profile, or other polymer matrices, liposomes or microspheres. They may also be formulated for rapid release, for example, lyophilized. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or any other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents, and may be of such a composition that they release the active ingredient only, or in a certain part of the digestive tract, optionally delayed. Examples of implantable compositions that may be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0050] The liquid dosage form for oral administration of the compound of the present disclosure includes pharmaceutically acceptable emulsion, microemulsion, solution, suspension, syrup and elixir.In addition to active ingredient, liquid dosage form may contain inert diluent commonly used in the art, such as water or other solvent, solubilizer and emulsifier, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerin, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid ester of sorbitan and their mixtures.

[0051] Besides inert diluents, the oral compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0052] Suspensions may contain, in addition to the active compounds, suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0053] IV. Treatment Method In one aspect, provided herein is a method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a derivative of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one or a salt thereof, in need of such treatment. In another aspect, provided herein is a method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a derivative of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one or a salt thereof, in need of such treatment, in need of such treatment.

[0054] In some embodiments, the method is a method of treating MS. In some embodiments, the method is a method of treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0055] In one aspect, a patient, e.g., a human patient, in need of treatment is administered an effective amount of a compound of formula Ia: [ka] or a salt thereof. In some embodiments, the compound of formula Ia or a salt thereof is about 90% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 95% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 98% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 99% pure.

[0056] In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula Ia or a salt thereof is a method of treating MS. In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula Ia or a salt thereof is a method of treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0057] In one aspect, a patient, e.g., a human patient, in need of treatment is administered an effective amount of a compound of formula Ib: [ka] or a salt thereof, wherein the compound or salt thereof is about 90% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 95% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 98% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 99% pure.

[0058] In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula Ib or a salt thereof is a method of treating MS. In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula Ib or a salt thereof is a method of treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0059] In one aspect, a patient, e.g., a human patient, in need of treatment is administered an effective amount of a compound of formula II: [ka] or a salt thereof, wherein the compound or salt thereof is about 99.6% pure.

[0060] In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula II or a salt thereof is a method of treating MS. In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula II or a salt thereof is a method of treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0061] In some embodiments, the compound is [ka] or a salt thereof, wherein the compound of formula IIa or a salt thereof is about 90% pure. In some embodiments, the compound of formula IIa or a salt thereof is about 95% pure. In some embodiments, the compound of formula IIa or a salt thereof is about 98% pure.

[0062] In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula IIa or a salt thereof is a method of treating MS. In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula IIa or a salt thereof is a method of treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0063] In some instances, the compound is [ka] or a salt thereof, wherein the compound of formula IIb or a salt thereof is about 90% pure. In some embodiments, the compound of formula IIb or a salt thereof is about 95% pure. In some embodiments, the compound of formula IIb or a salt thereof is about 98% pure.

[0064] In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula IIb or a salt thereof is a method of treating MS. In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, an effective amount of a compound of formula IIb or a salt thereof is a method of treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0065] In one aspect, the present invention provides a method for administering to a patient, e.g., a human patient, in need of treatment, a pharma- ceutical or pharmaceutical composition comprising administering to said patient ... [ka] and at least one compound or salt thereof selected from the group consisting of:

[0066] In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, a composition comprising a pharma- ceutically acceptable excipient and an effective amount of at least one compound selected from Formula Ia, Formula Ib, Formula II, and Formula III, or a salt thereof, is a method of treating MS. In some embodiments, the method of treatment comprising administering to a patient, e.g., a human patient, a composition comprising a pharma- ceutically acceptable excipient and an effective amount of at least one compound selected from Formula Ia, Formula Ib, Formula II, and Formula III, or a salt thereof, is a method of treating autoimmune disorders such as rheumatoid arthritis and lupus.

[0067] In one aspect, provided herein is a composition for use as a medicament for treating a patient in need of treatment, e.g., a human patient, comprising an effective amount of a derivative of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one or a salt thereof. In another aspect, provided herein is a composition for use as a medicament for treating a patient in need of treatment, e.g., a human patient, comprising a pharmaceutically acceptable excipient and an effective amount of a derivative of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one or a salt thereof.

[0068] In some embodiments, the medicament is for treating MS, hi some embodiments, the medicament is for treating autoimmune disorders such as rheumatoid arthritis and lupus.

[0069] In one aspect, provided herein is an effective amount of a compound of formula Ia: [ka] A composition is provided for use as a medicament for treating a patient, e.g., a human patient, in need of treatment, comprising a compound of formula Ia or a salt thereof. In some embodiments, the compound of formula Ia or a salt thereof is about 90% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 95% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 98% pure. In some embodiments, the compound of formula Ia or a salt thereof is about 99% pure.

[0070] In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment comprises an effective amount of a compound of formula Ia, or a salt thereof, for treating MS. In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment comprises an effective amount of a compound of formula Ia, or a salt thereof, for treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0071] In one aspect, provided herein is an effective amount of a compound of formula Ib: [ka] or a salt thereof, for use as a medicament for treating a patient, e.g., a human patient, in need of treatment, the composition comprising the compound of formula Ib or a salt thereof, wherein the compound or salt thereof is about 90% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 95% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 98% pure. In some embodiments, the compound of formula Ib or a salt thereof is about 99% pure.

[0072] In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment comprises an effective amount of a compound of formula Ia, or a salt thereof, for treating MS. In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment comprises an effective amount of a compound of formula Ia, or a salt thereof, for treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0073] In one aspect, provided herein is an effective amount of a compound of formula II: [ka] or a salt thereof, for use as a medicament for treating a patient, e.g., a human patient, in need of such treatment, wherein the compound or salt thereof is about 99.6% pure.

[0074] In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment comprises an effective amount of a compound of formula II or a salt thereof for treating MS. In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment comprises an effective amount of a compound of formula II or a salt thereof for treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0075] In some embodiments, the compound is [ka] or a salt thereof, wherein the compound of formula IIa or a salt thereof is about 90% pure. In some embodiments, the compound of formula IIa or a salt thereof is about 95% pure. In some embodiments, the compound of formula IIa or a salt thereof is about 98% pure.

[0076] In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment, comprises an effective amount of a compound of formula IIa, or a salt thereof, for treating MS. In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment, comprises an effective amount of a compound of formula IIa, or a salt thereof, for treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0077] In some instances, the compound is [ka] or a salt thereof, wherein the compound of formula IIb or a salt thereof is about 90% pure. In some embodiments, the compound of formula IIb or a salt thereof is about 95% pure. In some embodiments, the compound of formula IIb or a salt thereof is about 98% pure.

[0078] In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment, comprises an effective amount of a compound of formula IIb, or a salt thereof, for treating MS. In some embodiments, a composition for use as a medicament for treating a patient, e.g., a human patient, in need of treatment, comprises an effective amount of a compound of formula IIb, or a salt thereof, for treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0079] In one aspect, the present invention provides a method for the preparation of a pharmaceutical composition comprising a pharma- ceutically acceptable excipient and an effective amount of [ka] and at least one compound selected from the group consisting of:

[0080] In some embodiments, a composition for use as a medicament for treating a patient in need of treatment, e.g., a human patient, comprises a pharmaceutically acceptable excipient and an effective amount of at least one compound selected from Formula Ia, Formula Ib, Formula II, and Formula III, or a salt thereof, for treating MS. In some embodiments, a composition for use as a medicament for treating a patient in need of treatment, e.g., a human patient, comprises a pharmaceutically acceptable excipient and an effective amount of at least one compound selected from Formula Ia, Formula Ib, Formula II, and Formula III, and / or a salt thereof, for treating an autoimmune disorder, such as rheumatoid arthritis and lupus.

[0081] V. Synthesis method In one aspect, provided herein are methods for synthesizing and preparing derivatives of 4-amino-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one or salts thereof.

[0082] In one embodiment, the compound of formula Ia [ka] A method for preparing a compound of formula VIa or a salt thereof [ka] With acryloyl chloride.

[0083] In some embodiments, the method includes deprotecting in an acid system to form a compound of formula IX [ka] from the compound of formula VIa.

[0084] In one embodiment, the compound of formula Ib [ka] A method for preparing a compound of formula VIb or a salt thereof, comprising the steps of: [ka] With acryloyl chloride.

[0085] In some embodiments, the method includes deprotecting in an acid system to obtain a compound of formula VII. [ka] from the compound of formula VIb.

[0086] In some embodiments, the method includes deprotecting the compound of formula VIII using a base. [ka] from the compound of formula VII.

[0087] In some embodiments, the method uses triphenylphosphine and an azodicarboxylate to produce a compound of formula IX [ka] from the compound of formula VIII.

[0088] In some embodiments, the azodicarboxylate is selected from diethyl azodicarboxylate and diisopropyl azodicarboxylate. In some embodiments, the azodicarboxylate is diisopropyl azodicarboxylate.

[0089] In one embodiment, the compound of formula IV [ka] or a salt thereof, comprising reacting (4R)-5-amino-4-[4-amino-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]pentanoic acid with acryloyl chloride.

[0090] In some embodiments, the method further comprises preparing (4R)-5-amino-4-[4-amino-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]pentanoic acid or a salt thereof from (4R)-4-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(tert-butoxycarbonylamino)pentanoic acid by acid deprotection.

[0091] In some embodiments, the method further comprises preparing (4R)-4-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(tert-butoxycarbonylamino)pentanoic acid or a salt thereof from tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate by ring opening in the presence of a base.

[0092] In some embodiments, the method further comprises producing tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate or a salt thereof from tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate by electrochemical reaction utilizing a mediator and an electrolyte. In some embodiments, the mediator is quinucleidine. In further embodiments, the electrolyte is tetramethylammonium tetrafluoroborate.

[0093] In one embodiment, the compound of formula V [ka] or a salt thereof, comprising reacting 4-amino-1-[(1R)-1-(aminomethyl)-4-hydroxy-butyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one with acryloyl chloride.

[0094] In some embodiments, the method further comprises preparing 4-amino-1-[(1R)-1-(aminomethyl)-4-hydroxy-butyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one or a salt thereof from tert-butyl N-tert-butoxycarbonyl-N-[1-[(1R)-1-[(tert-butoxycarbonylamino)methyl]-4-hydroxy-butyl]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-4-yl]carbamate by acid deprotection.

[0095] In some embodiments, the method further comprises preparing tert-butyl N-tert-butoxycarbonyl-N-[1-[(1R)-1-[(tert-butoxycarbonylamino)methyl]-4-hydroxy-butyl]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-4-yl]carbamate, or a salt thereof, from tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate by ring opening in the presence of a base.

[0096] In some embodiments, the method further comprises producing tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate or a salt thereof from tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate by electrochemical reaction utilizing a mediator and an electrolyte. In some embodiments, the mediator is quinucleidine. In further embodiments, the electrolyte is tetramethylammonium tetrafluoroborate.

[0097] [Table 1] EXAMPLES

[0098] Synthesis of 4-amino-1-[(3R,5S)-5-hydroxy-1-prop-2-enoyl-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one and 4-amino-1-[(3R,5R)-5-hydroxy-1-prop-2-enoyl-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one

[0099] [ka]

[0100] Compounds of Formula Ia and Formula Ib were synthesized as shown in the scheme above (designated Compound 13 and Compound 14, respectively) and as described in more detail below.

[0101] Step 1: Under nitrogen atmosphere, a reactor was charged with DMF (21.5 mL). 2,4-Dichloro-3-nitropyridine (SM1) (4.3 g, 22.28 mmol, 1.0 equiv) was added at 20±5° C. After stirring for 5 min, triethylamine (4.75 mL, 33.81 mmol, 1.5 equiv) was added and the mixture was cooled to 0±5° C. HOBt (301 mg, 2.23 mmol, 0.1 equiv) was added followed by a solution of tert-butyl (3R,5S)-3-amino-5-hydroxy-piperidine-1-carboxylate (4.8 g, 22.28 mmol, 1.0 equiv) in DMF (64.5 mL) added dropwise. The temperature was maintained between −10° C. and 0° C. during the addition of the reagents. The reaction mixture was stirred at 0±5° C. for 2 h and then at 20±5° C. for 4 h. HPLC analysis confirmed the production of tert-butyl (3R,5S)-3-[[2-(benzotriazol-1-yloxy)-3-nitro-4-pyridyl]amino]-5-hydroxy-piperidine-1-carboxylate (110).

[0102] Step 2: The next step was carried out immediately in one pot by charging triethylamine (4.75 mL, 33.81 mmol, 1.5 equiv) and 1-(4-methoxyphenyl)-N-[(4-methoxyphenyl)methyl]methanamine (5.7 g, 22.28 mmol, 1.0 equiv). The reaction solution was warmed to 40-45 °C and stirred at this temperature for 62 h. HPLC analysis confirmed that less than 1% intermediate remained. After cooling to 10-20 °C, EtOAc (129 mL) and water (129 mL) were added to the solution and stirred at 10-20 °C for 30 min. The organic phase was separated and washed with aqueous CaCl2 (5%, 43 mL x 5). The organic phase was concentrated under reduced pressure below 35° C. to give a solution of tert-butyl (3R,5S)-3-[[2-[bis[(4-methoxyphenyl)methyl]amino]-3-nitro-4-pyridyl]amino]-5-hydroxy-piperidine-1-carboxylate (111) (94.3% purity) in EtOAc (brown solution), which was used directly in the next step.

[0103] Step 3: tert-Butyl (3R,5S)-3-[[2-[bis[(4-methoxyphenyl)methyl]amino]-3-nitro-4-pyridyl]amino]-5-hydroxy-piperidine-1-carboxylate (111) (14.7 g, 24.76 mmol, 1 equiv.) (theoretical amount) was dissolved in AcOEt (73.5 mL) and charged to a reactor. The reactor was inerted under vacuum and filled with nitrogen 5 times, wet 10% Pd / C (2.64 g, 20 wt.%) was added, the reactor was inerted under vacuum and filled with nitrogen 5 times, then the reactor was inerted under vacuum and filled with hydrogen 3 times. The reaction was kept under hydrogen pressure at 20±5° C. and stirred for 22 h. HPLC analysis confirmed the disappearance of starting material. The suspension was filtered and the cake was washed with EtOAc (73.5 mL×5). The filtrate was concentrated under reduced pressure to give tert-butyl (3R,5S)-3-[[3-amino-2-[bis[(4-methoxyphenyl)methyl]amino]-4-pyridyl]amino]-5-hydroxy-piperidine-1-carboxylate (112) as a yellow foamy solid (12 g, yield=87%, purity 91.1%), which was used in the next step without further purification.

[0104] Step 4: A reactor was charged with THF (60 mL), tert-butyl (3R,5S)-3-[[2-[bis[(4-methoxyphenyl)methyl]amino]-3-nitro-4-pyridyl]amino]-5-hydroxy-piperidine-1-carboxylate (112) (12.0 g, 20.21 mmol, 1.0 equiv.) and triethylamine (4.4 mL, 40.42 mmol, 2.0 equiv.). The solution was stirred at 20±5°C for 10 min, then triphosgene (1.07 mL, 6.46 mmol, 0.32 equiv.) was added in three portions and the resulting solution was stirred at 20-25°C for 1.5 h. HPLC analysis confirmed that less than 1% intermediate remained. The solution was concentrated to 24 mL, CHCl (120 mL) and aqueous NaHCO (60 mL) were added and the mixture was stirred for 10 min. The organic phase was separated and concentrated under reduced pressure below 40 °C to give the crude product, which was purified by SGC (CHCl / MeOH = 30 / 1) to give tert-butyl (3R,5S)-3-[4-[bis[(4-methoxyphenyl)methyl]amino]-2-oxo-3H-imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (113) as a foamy solid (10 g, yield = 74%, purity 97.1%).

[0105] Step 5: A solution of tert-butyl (3R,5S)-3-[4-[bis[(4-methoxyphenyl)methyl]amino]-2-oxo-3H-imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (113) (10.0 g, 16.95 mmol, 1.0 equiv) in CHCl (70 mL) was stirred at 20±5° C. for 10 min, then cooled to 5±5° C.; TFA (10 mL) was added slowly, maintaining the temperature below 10° C. The reaction solution was stirred at 5±5° C. for 30 min, warmed to 40±5° C., and then stirred for 14 h. HPLC analysis confirmed less than 1.0% starting material. The reaction was cooled to 15-25° C. The pH was adjusted to 9-10 with aqueous NaCO. The aqueous phase containing 4-amino-1-[(3R,5S)-5-hydroxy-3-piperidyl]-3H-imidazo[4,5-c]pyridin-2-one (117) was separated and saved for direct use in the next step.

[0106] Step 6: To an aqueous solution containing 4-amino-1-[(3R,5S)-5-hydroxy-3-piperidyl]-3H-imidazo[4,5-c]pyridin-2-one (117) cooled to 5±5°C, CHCl (40 mL) was added and a solution of BocO (3.7 g, 16.95 mmol, 1.0 equiv) in CHCl (10 mL) was added dropwise at 5±5°C. The solution was warmed to 15-25°C and stirred at 20-25°C for 42 h. HPLC analysis confirmed that the starting material was less than 3.0%. The aqueous phase was separated and extracted with CHCl (100 mL x 3) and THF (100 mL x 2). The organic phases were combined and concentrated under reduced pressure below 35 °C to give 5.4 g of tert-butyl (3R,5S)-3-(4-amino-2-oxo-3H-imidazo[4,5-c]pyridin-1-yl)-5-hydroxy-piperidine-1-carboxylate (118) as a solid with a purity of 92.2% (containing CH2Cl2 / THF).

[0107] Step 7: A reactor was charged with CHCl (21.6 mL) and tert-butyl (3R,5S)-3-(4-amino-2-oxo-3H-imidazo[4,5-c]pyridin-1-yl)-5-hydroxy-piperidine-1-carboxylate (118) (5.4 g, 15.45 mmol, 1.0 equiv). The suspension was stirred for 10 min. After addition of N,N-dimethylformamide dimethyl acetal (6.1 mL, 46.35 mmol, 3.0 equiv) at 20±5° C., the solution became clear. After stirring for 20 h at 20±5° C., a solid precipitated. HPLC analysis confirmed less than 2.0% starting material. MTBE (54 mL) was added and the reaction was stirred for 2 h at 20±5° C. The suspension was filtered. The cake was washed with MTBE (21.6 mL) and dried under reduced pressure at 35±5° C. for 8 h to give tert-butyl (3R,5S)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3H-imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (7) as an off-white solid (4.3 g, yield=83%, purity 93.4%).

[0108] Step 8: At 20±5° C., a reactor was charged with CHCl (75 mL), Cu(OAc) (561.2 mg, 3.09 mmol, 0.5 equiv), 2,2′-dipyridyl (482 mg, 3.09 mmol, 0.5 equiv), CsCO (4.02 g, 12.36 mmol, 2.0 equiv), 4 Å MS (2.5 g, 1.0 w / w) and tert-butyl (3R,5S)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3H-imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (7) (2.5 g, 6.18 mmol, 1.0 equiv). Compressed air (21% oxygen and 79% nitrogen) was bubbled through the reaction for 30 min at 20 ± 5 °C. 4-Phenoxyphenylboronic (1.98 g, 9.27 mmol, 1.5 equiv) was added to the reaction mixture in four portions over 1.5 h at 15-25 °C. Compressed air (21% oxygen and 79% nitrogen) was bubbled through the reaction for 5 h at 20 ± 5 °C. HPLC analysis confirmed less than 5.0% starting material. The suspension was filtered and the cake was washed with CHCl (25 mL × 2). The filtrate was concentrated and the crude product was purified by SGC (CHCl / MeOH=50 / 1) to give tert-butyl (3R,5S)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (compound of formula IX, also referred to above as (8)) as a solid in 97.5% purity and 47.2% yield.

[0109] Step 9: tert-Butyl (3R,5S)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (8) (0.3 g, 0.52 mmol, 1.0 equiv.) was dissolved in CHCl (1.5 mL). To this solution, aqueous HCl (3N, 3 mL) was added dropwise at 20±5°C. The solution was stirred at 30-35°C for 2 h and at 60-65°C for 14 h. HPLC analysis confirmed that the starting material was less than 1.0%. To the solution cooled to 20-25°C, CHCl (15 mL) and water (45 mL) were added. After the solution was stirred for 10 min, the aqueous phase was separated and extracted with CHCl (30 mL) and the pH of the aqueous phase was adjusted to 9-10 with aqueous NaCO. The aqueous phase turned pink in color. After stirring for 30 min, the organic phase was extracted, washed with brine (15 mL), dried over NaSO, and filtered to give a solution of 4-amino-1-[(3R,5S)-5-hydroxy-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (compound of formula VIa, also referred to above as (11A)) in CHCl (81% purity), which was used directly in step 10.

[0110] Step 10: The previous solution of 4-amino-1-[(3R,5S)-5-hydroxy-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (11A) (164 mg, 0.39 mmol, 1 equiv.) in CHCl was cooled to 0±5° C., N,N-diisopropylethylamine (133 μL, 0.78 mmol, 2.0 equiv.) and trimethylsilyl chloride (50 μL, 0.39 mmol, 1.0 equiv.) in CHCl (2.5 ml) were charged, and the solution was stirred for 30 min at 0±5° C. The solution was cooled to −20±5° C., followed by dropwise addition of acryloyl chloride (32 μL, 0.39 mmol, 1.0 equiv.) in CHCl (5 ml). The reaction was quenched with water (20 mL), and the organic phase was separated and concentrated under reduced pressure below 35° C. to give the crude product. The crude product was purified by preparative TLC (CHCl / EtOH=15 / 1), the fractions were concentrated, and CHCl was exchanged for EtOH. The solution was concentrated to 10 mL and n-heptane (15 ml) was charged. The solution was then concentrated to 10 mL to allow the solid to precipitate. The precipitate was filtered to give 80 mg of 4-amino-1-[(3R,5S)-5-hydroxy-1-prop-2-enoyl-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (compound of formula Ia, also referred to above as (13)) as an off-white solid with 98.1% purity and 43% yield. 1H NMR spectrum (500 MHz, DMSO D6, δ in ppm): 7.77 (d, 1H); 7.51-7.41 (m, 4H); 7.22 (t, 1H); 7.18-7.11 (m, 4H); 6.96 (br. m, 1H); 6.81 (m, 1H); 6.15 (d, 1H); 5.71 (dd, 1H); 5.28 (br. s, 1H); 4.83 (br. m, 2H); 4.58 (br. d, 0.5 H); 4.47 (br. d, 0.5 H); 4.25 (br. qd, 1H); 4.15 (br. d, 0.5 H); 4.1 (br. d, 0.5 H); 3.69 (br. t, 0.5 H); 3.60 (m, 1H); 3.27 (br. t, 0.5 H); 2.95 (br. t, 0.5 H); 2.49 (br. t, 0.5 H); 2.28 (m, 1H); 2.13 (br. m, 1H)

[0111] Step 9B: An azote deactivated reactor was charged with THF (12.5 mL), DIAD (683 μL, 3.48 mmol, 4.0 equiv), and Ph3P (912 mg, 3.48 mmol, 4.0 equiv). The reaction was cooled to 0-5 °C, after which tert-butyl (3R,5S)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (8) (0.5 g, 0.87 mmol, 1.0 equiv) was added to the reactor. The reaction was stirred at 0-5°C for 10 min, 4-nitrobenzoic acid (291 mg, 1.74 mmol, 2.0 equiv) was added to the reactor, and the reaction was stirred at 0-5°C for an additional 2 h. The reaction mixture was then stirred at 15-20°C for 14 h and concentrated under reduced pressure below 35°C. The residue was purified by SGC (CHCl / MeOH=50 / 1) to give crude tert-butyl (3R,5R)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(4-nitrobenzoyl)oxy-piperidine-1-carboxylate (compound of formula VIII, also referred to above as (9B)) as a foamy solid (87% purity) containing PhPO. PhPO was difficult to separate, and this intermediate was not further purified.

[0112] Step 10B: A flask was charged with MeOH (9 mL), tert-butyl (3R,5R)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(4-nitrobenzoyl)oxy-piperidine-1-carboxylate (9B) (0.45 g, 0.62 mmol, 1.0 equiv.) and K2CO3 (258 mg, 1.87 mmol, 3.0 equiv.). The reaction mixture was stirred at 15-20 °C for 14 h. HPLC analysis showed no starting material remained. The suspension was filtered, the cake washed with MeOH (2.25 mL) and the filtrate was concentrated under reduced pressure below 35 °C to give the crude product. The crude product was purified by SGC (CHCl / MeOH=50 / 1) to give tert-butyl (3R,5R)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (compound of formula VII, also referred to above as (10B)) (0.16 g, yield=44%, purity 98.2%) as a powder solid.

[0113] Step 11B: A flask was charged with CHCl (1 mL), tert-butyl (3R,5R)-3-[4-[(Z)-dimethylaminomethyleneamino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-piperidine-1-carboxylate (10B) (0.16 g, 0.28 mmol, 1 equiv.) and aqueous HCl (3.0 M, 2 mL). The solution was stirred at 25-30° C. for 2 h and then at 60-65° C. for 14 h. HPLC analysis showed no starting material remained. The reaction mixture was cooled to 15-20° C. and CHCl (15 ml) and H0 (30 ml) were added to the flask. The organic phase was separated. CHCl (30 ml) was added to the aqueous phase, and the pH value of the aqueous phase was adjusted to 9-10 with saturated aqueous NaCO. The organic phase was separated, and the aqueous phase was re-extracted with CHCl (30 mL×2). The different organic phases were collected, washed with brine (10 mL), dried over NaSO, filtered, and concentrated under reduced pressure below 35° C. to give 0.11 g of the crude material 4-amino-1-[(3R,5R)-5-hydroxy-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (compound of formula VIb, also referred to above as (11B)) as a foamy solid, which was used directly in step 12B without further purification.

[0114] Step 12B: To a solution of 4-amino-1-[(3R,5R)-5-hydroxy-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (11B) (0.11 g, 0.26 mmol, 1 equiv.) in CHCl (44 mL) was added N,N-diisopropylethylamine (177 μL, 1.04 mmol, 4.0 equiv.). The solution was cooled to -5 to 0 °C and a solution of trimethylsilyl chloride (33 μL, 0.26 mmol, 1.0 equiv.) in CHCl (2.5 ml) was added dropwise. The solution was stirred at -5 to 0 °C for 30 min and cooled to -25 to 20 °C. Acryloyl chloride (27.5 μL, 0.34 mmol, 1.3 equiv) in CHCl (2.5 mL) was added dropwise. The reaction was stirred for 10 min, warmed to 0-5° C., and quenched with water (20 mL). The organic phase was separated, washed with brine (10 mL), and concentrated under reduced pressure below 35° C. to give the crude material. The crude product was purified by preparative TLC (CHCl / MeOH=15 / 1) to give 4-amino-1-[(3R,5R)-5-hydroxy-1-prop-2-enoyl-3-piperidyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (compound of formula Ib, also referred to above as (14)) (63 mg, yield=51%, purity 99.1%) as an off-white powder solid. LC / HRMS: 472.1821 (M+H) + , 470.1790 (MH) - . 1H NMR spectrum (500 MHz, DMSO D6, δ in ppm): 7.76 (d, 1H); 7.50-7.42 (m, 4H); 7.22 (t, 1H), 7.18-7.11 (m, 4H); 6.90 (d, 1H); 6.79 (dd, 1H); 6.13 (dd, 1H); 5.68 (dd, 1H); 5.05 (br. d, 0.75 H); 4.96 (br. d, 0.25 H); 4.81 (br. s, 2H); 4.65 (br. m, 0.25 H); 4.59-4.46 (m, 1.75 H); 4.19 (br. d, 0.25 H); 4.07 (br. s, 1H); 3.97 (br. d, 0.75 H); 3.72 (t, 0.25 H); 3.38 (d, 0.75 H); 3.32 (m, 0.75H); 2.90 (d, 0.25 H); 2.53 (m, 1H); 1.92 (br. d, 1H). EXAMPLES

[0115] Synthesis of 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-1-(2,3-dihydroxypropanoyl)-3-piperidyl]imidazo[4,5-c]pyridin-2-one [ka]

[0116] As shown in the above scheme, compounds of formula IIa and IIb were prepared by the reaction of AD-mix-alpha (i.e., (DHQ)2PHAL (hydroquinine 1,4-phthalazinediyl diether, Sigma Aldrich Cat. No. 392723) 0.0016 mol; potassium carbonate, powder 0.4988 mol; potassium ferricyanide 0.4988 mol; potassium osmate dihydrate 0.0007 mol) or AD-mix-beta (i.e., (DHQD)2PHAL (Sigma Aldrich Cat. No. 392723) 0.0016 mol; potassium carbonate, powder 0.4988 mol; potassium ferricyanide 0.4988 mol; potassium osmate dihydrate 0.0007 mol). Prepared from 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-1-prop-2-enoyl-3-piperidyl]imidazo[4,5-c]pyridin-2-one using: Aldrich catalog number 392731 (0.0016 mol); potassium carbonate, powder 0.4988 mol; potassium ferricyanide 0.4988 mol; potassium osmate dihydrate 0.0007 mol).

[0117] The starting material, 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-1-prop-2-enoyl-3-piperidyl]imidazo[4,5-c]pyridin-2-one, was prepared according to the procedure described in Example 3 of U.S. Pat. No. 9,688,676.

[0118] To prepare compounds of formula IIa and IIb, a suspension of AD-mix-alpha (154 mg) or AD-mix-beta (154 mg) in water / tBuOH (1 / 1, 1 mL / 1 mL) was stirred for about 10 min until the solid dissolved. Under argon atmosphere, the starting material 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-1-prop-2-enoyl-3-piperidyl]imidazo[4,5-c]pyridin-2-one was added to the yellow solution. The starting material was insoluble in the mixture, resulting in a yellow suspension. The suspension was stirred at room temperature for 20 h. The progress of the reaction was monitored by TLC (AcOEt / MeOH 10 / 1). The reaction mixture was hydrolyzed with Na2SO3, stirred for 20 min, and extracted with CH2Cl2. The organic phase was separated, dried over MgSO4, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by SCX (strong cation exchange) column (MeOH, then 3.5N solution of NH3 in MeOH) to give a clear oil, 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-1-(2,3-dihydroxypropanoyl)-3-piperidyl]imidazo[4,5-c]pyridin-2-one, as a mixture of two diastereoisomers separated by chiral chromatography to give the compound of formula IIa (98% pure) and the compound of formula IIb (98% pure).IIa: 1H NMR (500 MHz, DMSO-d6) (2 rotamers 50 / 50) δ ppm 1.45 - 1.65 (m, 1 H), 1.80 - 1.95 (m, 2 H), 2.45 (m, 1 H), 3.10 (m, 0.5 H), 3.25 (m, 0.5 H), 3.40 - 3.55 (m, 2 H), 3.67 (m, 0.5 H), 4.05 - 4.50 (m, 4.5 H), 4.62 (t, J = 4 Hz, 0.5 H), 4.77 (t, J = 4 Hz, 0.5 H), 4.80 (m, 2 H), 4.95 (d, J = 4 Hz, 0.5 H), 5.01 (d, J = 4 Hz, 0.5 H), 6.90 - 6.96 (m, 1 H), 7.15 (m, 4 H), 7.22 (t, J = 7 Hz, 1 H), 7.45 (m, 4 H), 7.75 (d, J = 5 Hz, 1 H). IIb: 1H NMR (500 MHz, DMSO-d6) (2 rotamers 50 / 50) δ ppm 1.45 - 1.65 (m, 1 H), 1.80 - 1.95 (m, 2 H), 2.45 (m, 1 H), 3.10 (m, 0.5 H), 3.25 (m, 0.5 H), 3.40 - 3.55 (m, 2 H), 3.67 (m, 0.5 H), 4.05 - 4.50 (m, 4.5 H), 4.65 (t, J = 4 Hz, 0.5 H), 4.76 (t, J = 4 Hz, 0.5 H), 4.80 (m, 2 H), 4.98 (m, 1 H), 6.90 - 6.96 (m, 1 H), 7.15 (m, 4 H), 7.22 (t, J = 7 Hz, 1 H), 7.45 (m, 4 H), 7.75 (d, J = 5 Hz, 1 H) LC / MS: 490 (M + H) + .

Example

[0119] Synthesis of (R)-5-acrylamido-4-(4-amino-2-oxo-3-(4-phenoxyphenyl)-2,3-dihydro-1H-imidazo[4,5-c]pyridin-1-yl)pentanoic acid [ka]

[0120] The compound of formula IV (referred to above as compound (6)) was prepared as shown in the scheme above and as described in more detail below. The starting material, 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-3-piperidyl]imidazo[4,5-c]pyridin-2-one (1), was prepared according to the procedure described in Example 2 of U.S. Pat. No. 9,688,676. 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-3-piperidyl]imidazo[4,5-c]pyridin-2-one (1) was then converted to the oxalate salt by adding DCM to compound (1). The solution was exchanged with EtOH and the resulting solution was reacted with oxalic acid. After filtration, the crude solid was slurried in MTBE, filtered, and dried to obtain the oxalate salt of compound (1).

[0121] Step 1: Under argon atmosphere, a flask was charged with CHCl and the oxalate salt of 4-amino-3-(4-phenoxyphenyl)-1-[(3R)-3-piperidyl]imidazo[4,5-c]pyridin-2-one (1.3 g, 2.66 mmol, 1 eq.). To this suspension was then added DMAP (487 mg, 3.98 mmol, 1.5 eq.), DIEA (1.35 mL, 7.97 mmol, 3 eq.) and BocO (2.90 g, 13.28 mmol, 5 eq.). The starting material dissolved and the reaction became clear. The solution was stirred at room temperature for 20 h. The reaction was hydrolyzed with water and extracted with CHCl. ​​The organic phase was separated, dried over MgSO, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by flash chromatography (SiO2, Hep / AcOEt (1 / 1)) to give tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate (2) (1.86 g, yield=91.2%).

[0122] Step 2: To a solution of tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate (2) (1 g, 1.42 mmol, 1 equiv.) in MeCN / HO / CCl (15 mL / 20 mL / 15 mL) was added NaIO (1.22 g, 5.7 mmol, 4 equiv.) and RuCl.xHO (32 mg, 142 μmol, 0.1 equiv.) in sequence. The reaction was a light brown mixture. After stirring at room temperature for 1 h, additional NaIO (152 mg, 0.7 mmol, 0.25 equiv.) was added. The reaction was stirred at room temperature for 24 h, at which point it turned dark brown. After hydrolysis with aqueous Na2S2O5 and stirring for 30 min, the solution turned pale yellow, which was extracted with AcOEt, the organic phase was separated, dried over MgSO4, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by flash chromatography (SiO2, Hep / AcOEt (1 / 1)) to give tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate (starting material) (450 mg, yield=45%), and tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate (desired product) (3) (200 mg, yield=20%).

[0123] Step 2: Electrochemical process - Using an Electrasyn 2.0 (IKA) equipped with a 20 mL vial setup, to a solution of tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate (2) (300 mg, 0.43 mmol, 1 equiv.) in MeCN (12 mL) was added quinuclidine (2 equiv., 0.86 mmol, 95 mg) as a mediator, 1,1,1,3,3,3-hexafluoropropan-2-ol ("HFIP") (11 equiv., 4.70 mmol, 790 mg), and tetramethylammonium tetrafluoroborate (2.05 equiv., 0.88 mmol, 141 mg) as an electrolyte. Vial cap set: (working electrode) RVC-Reticulated Vitreous Carbon Foam was used as the anode and Ni Foam was used as the cathode. The electrochemical reaction was carried out in constant current mode; a constant current of 8 mA without polarity switching. The total charge was 16 Faradays / mol at room temperature. The reaction was monitored by LC / MS. After the reaction was poured into ice water, a 1M solution of sodium thiosulfate was added. The aqueous phase was washed with ethyl acetate. The organic phase was separated, dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Purification by flash chromatography on silica gel, eluting with ethyl acetate / pentane (70 / 30) gave the expected product tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate (3) (150 mg, yield=49%).

[0124] Step 3: To a solution of tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate (3) (820 mg, 1.15 mmol, 1 equiv.) in THF / H2O (30 mL / 30 mL) was added LiOH (471 mg, 19.4 mmol, 17 equiv.). The solution was stirred at room temperature for 3 h. The reaction mixture was hydrolyzed with 1N aqueous HCl until pH=1-2 and then extracted with AcOEt. The organic phase was separated, dried over MgSO4, filtered and concentrated under reduced pressure to give the desired compound: (4R)-4-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(tert-butoxycarbonylamino)pentanoic acid (4) (780 mg, yield=92.8%).

[0125] Step 4: Under an argon atmosphere, a flask was charged with CHCl (5 mL), (4R)-4-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(tert-butoxycarbonylamino)pentanoic acid (720 mg, 0.98 mmol, 1 equiv.) (4), and TFA (2 mL). The mixture was stirred at room temperature for 6 h. After evaporating CHCl and TFA and drying under high vacuum for 10 h, (4R)-5-amino-4-[4-amino-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]pentanoic acid (5) (410 mg, 96%) was obtained, which was used directly in the next step without further purification.

[0126] Step 5: To a suspension of (4R)-5-amino-4-[4-amino-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]pentanoic acid (5) (425 mg, 0.98 mmol, 1 equiv.) and CHCl (6 mL) was added DMF (0.5 mL) to solubilize the starting material. The reaction solution was then cooled to 0° C. and charged with DIEA (343 μL, 1.96 mmol, 2 equiv.) and a solution of acryloyl chloride (41 μL, 0.49 mmol, 0.5 equiv.) in CHCl (2 mL) was added dropwise. The solution was allowed to warm to room temperature and stirred for 2 h. The reaction was concentrated under reduced pressure and purified directly by preparative HPLC followed by supercritical HPLC to give (4R)-4-[4-amino-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-(prop-2-enoylamino)pentanoic acid (compound of formula IV, also referred to above as (6)) (180 mg, yield=38%, purity 98%). 1H NMR (500 MHz, DMSO-d6) δ ppm 2.02 (m, 1 H), 2.17 (m, 2 H), 2.28 (m, 1 H), 3.65 (m, 2 H), 4.36 (m, 1 H), 4.78 (s, 2 H), 5.53 (dd, J=8 , 2Hz , 1 H), 6.02 (dd, J=17 , 2 Hz, 1 H), 6.09 (dd, J=17 , 8 Hz ,1 H), 6.65 (m, 1 H), 7.15 (d, J=8 Hz, 4 H), 7.21 (t, J=7 Hz, 1 H), 7.45 (m, 4 H), 7.71 (d, J=5 Hz, 1 H), 8.28 (t, J=6 Hz, 1H) LC / MS: 488.2 (M+H) + / 486.1 (MH) - . EXAMPLES

[0127] Synthesis of (R)-N-(2-(4-amino-2-oxo-3-(4-phenoxyphenyl)-2,3-dihydro-1H-imidazo[4,5-c]pyridin-1-yl)-5-hydroxypentyl)acrylamide [ka]

[0128] The compound of formula V (referred to above as compound (6)) was prepared as shown in the scheme above and as detailed below.

[0129] Steps 1 and 2 were carried out as described in steps 1 and 2 of Example 3. Step 3: At room temperature and under argon atmosphere, a flask was charged with tert-butyl (5R)-5-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-2-oxo-piperidine-1-carboxylate (3) (280 mg, 0.39 mmol, 1 eq.), MeOH (8 mL), and NaBH4 (148 mg, 3.9 mmol, 10 eq.). The reaction mixture was stirred at room temperature for 3 h and then hydrolyzed with H2O. The organic phase was separated, dried over MgSO4, filtered, and concentrated under reduced pressure to give the desired compound tert-butyl (3R)-3-[4-[bis(tert-butoxycarbonyl)amino]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]piperidine-1-carboxylate (4) (281 mg, yield=77%), which was used directly in the next step without further purification.

[0130] Step 4: To the starting material tert-butyl N-tert-butoxycarbonyl-N-[1-[(1R)-1-[(tert-butoxycarbonylamino)methyl]-4-hydroxy-butyl]-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-4-yl]carbamate (4) (216 mg, 0.30 mmol, 1 eq.) was added HCl in dioxane (15 mL). The solution was stirred at room temperature for 2 h. The solvent was evaporated and the crude product 4-amino-1-[(1R)-1-(aminomethyl)-4-hydroxy-butyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (5) (126 mg, yield=95.3%) was used in the next step.

[0131] Step 5: Under an argon atmosphere, a flask was charged with 4-amino-1-[(1R)-1-(aminomethyl)-4-hydroxy-butyl]-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-2-one (125 mg, 0.29 mmol, 1 eq) (5) and CHCl (2 mL). DIEA (104 μL, 0.59 mmol, 2 eq) was then charged, after which the reaction solution was cooled to 0° C. and a solution of acryloyl chloride (12.5 μL, 0.15 mmol, 0.5 eq) in CHCl (1 mL) was added dropwise. The solution was kept at 0° C. and stirred for 1 h. The reaction was hydrolyzed with H0 and extracted with AcOEt. The organic phase was separated, dried over MgSO4, filtered and concentrated under reduced pressure to give the crude compound which was purified by preparative HPLC to give N-[(2R)-2-[4-amino-2-oxo-3-(4-phenoxyphenyl)imidazo[4,5-c]pyridin-1-yl]-5-hydroxy-pentyl]prop-2-enamide (compound of formula V, referred to above as (6)) (38 mg, yield=27%, purity>95%). 1H NMR (500 MHz, DMSO-d6) δ ppm 1.25 - 1.40 (m, 2 H), 1.83 (m, 1 H), 2.08 (m, 1 H), 3.35 (m, 2 H), 3.50 - 3.70 (m, 2 H), 4.35 (m, 1 H), 4.43 (m, 1 H), 4.80 (s, 2 H), 5.53 (dd, J=8 , 2Hz , 1 H), 6.03 (dd, J=17 , 2 Hz, 1 H), 6.11 (dd, J=17 , 8 Hz ,1 H), 6.70 (m, 1 H), 7.15 (d, J=8 Hz, 4 H), 7.22 (t, J=7 Hz, 1 H), 7.43 (d, J=7 Hz, 2 H), 7.48 (t, J=7 Hz, 2 H), 7.74 (d, J=5 Hz, 1 H), 8.29 (t, J=6 Hz, 1H) LC / MS: 474.1 (M+H) + . EXAMPLES

[0132] BTK(h) Test Compounds of Formula Ib, Formula II, and Formula III were tested against BTK(h) as described herein. All compounds tested were made into working stocks in 100% DMSO at 50x the final assay concentration. If necessary, more concentrated stocks were manually diluted 50x with 100% DMSO. Powdered compounds were reconstituted to 10 mM stocks in 100% DMSO before further dilution 50x.

[0133] Assay procedure: 50x the required amount of test compound stock was added to the assay wells prior to the addition of the reaction mix containing enzyme and substrate. The reaction was initiated by the addition of the selected concentration of ATP. There was no pre-incubation of the enzyme / substrate mix with compound prior to the addition of ATP.

[0134] Data Analysis: Results are expressed as percent remaining kinase activity relative to DMSO control, which is calculated using the following formula:

number

[0135] I C 50 Data are analyzed using XLFit version 5.3 (ID Business Solutions) for determination of R. Nonlinear regression analysis is used to fit sigmoidal dose-response (variable slope) curves based on the average results for each test concentration. If the top and / or bottom ends of the curve deviate by more than 10% from 100 and 0, respectively, an R 2 Either or both of these limits can be 100 and 0, provided that QC criteria for

[0136] The results are shown in Table 2.

[0137] [Table 2] EXAMPLES

[0138] In vitro testing Compounds of formula Ia, formula Ib, formula III, formula IV, and formula V were tested in various in vitro assays. Compound binding was calculated as the % inhibition of the binding of radiolabeled ligand specific to each target. Enzyme inhibition effect of compounds was calculated as the % inhibition of control enzyme activity. Certain compounds showed significant activity in certain assays, defined in this example as more than 50% inhibition or stimulation.

[0139] BZD binding assay: The general procedure is described below. See Le Fur, G. et al. (1983), Life Sci., 33:449-457. Kd is the affinity of the radioligand for the receptor. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0140] [Table 3]

[0141] The compound of formula Ia showed significant activity in this assay, exhibiting 52.2% inhibition.

[0142] Dopamine binding assay: The general procedure is described below. See Pristupa, ZB et al. (1994), Mol. Pharmacol., 45:125-135. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0143] [Table 4]

[0144] Compound of formula Ia showed significant activity in this assay, exhibiting 65% inhibition. Compound of formula III also showed significant activity, exhibiting 96.2% inhibition.

[0145] D1(h) binding assay: The general procedure is described below. See Zhou, QY et al. (1990), Nature, 347:76-80. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0146] [Table 5]

[0147] The compound of formula III showed significant activity in this assay, exhibiting 80% inhibition.

[0148] H2(h) Binding Assay: The general procedure is described below. See Leurs, R. et al. (1994), Brit. J. Pharmacol., 112:847-854. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0149] [Table 6]

[0150] The compound of formula III showed significant activity in this assay, exhibiting 56.7% inhibition.

[0151] δ(DOP)(h) Binding Assay: The general procedure is described below. See Simonin, F. et al. (1994), Mol. Pharmacol., 46:1015-1021. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0152] [Table 7]

[0153] The compound of formula III showed significant activity in this assay, exhibiting 56.7% inhibition.

[0154] Kappa (h) (KOP) binding assay: The general procedure is described below. See Simonin, F. et al. (1995), Proc. Natl. Acad. Sci. USA, 92:7006-7010. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0155] [Table 8]

[0156] The compound of formula III showed significant activity in this assay, exhibiting 54.6% inhibition.

[0157] 5-HT 5a (h) Binding Assay: The general procedure is described below. See Rees, S. et al. (1994), FEBS Lett., 355:242-246. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0158] [Table 9]

[0159] The compound of formula III showed significant activity in this assay, exhibiting 56.1% inhibition.

[0160] Norepinephrine Transporter (h) Binding Assay: The general procedure is described below. See Pacholczyk, T. et al. (1991), Nature, 350:350-354. Minor changes to the experimental protocol described below may have occurred during testing, but if so, did not affect the quality of the results obtained.

[0161] [Table 10]

[0162] The compound of formula III showed significant activity in this assay, exhibiting 66.7% inhibition.

[0163] Equivalent The above specification is believed to be sufficient to enable those skilled in the art to practice the embodiments. The above description and examples detail certain embodiments and describe the best mode contemplated by the inventors. However, no matter how detailed the above appears in the text, it will be understood that the embodiments can be practiced in many ways and must be construed in accordance with the appended claims and any equivalents thereof.

[0164] As used herein, the term about refers to numerical values, including, for example, integers, decimals, and percentages, whether or not explicitly stated. The term about generally refers to a range of numerical values ​​(e.g., + / - 5 to 10% of the recited range) that one of ordinary skill in the art would consider equivalent to the recited value (e.g., having the same function or result). When the terms at least and about precede a recitation of a numerical value or range, they modify all of the values ​​or ranges provided in that recitation. In some cases, the term about may include numerical values ​​that are rounded to the nearest significant figure.

Claims

1. Formula Ia: 【Chemistry 1】 or a salt thereof, wherein the compound or salt thereof has a purity of about 90%.

2. 10. The compound of claim 1, wherein the compound or a salt thereof is about 95% pure.

3. 10. The compound of claim 1, wherein the compound or a salt thereof is about 98% pure.

4. Formula Ib: 【Chemistry 2】 or a salt thereof, wherein the compound or salt thereof has a purity of about 90%.

5. 5. The compound of claim 4, wherein the compound or salt thereof is about 95% pure.

6. 5. The compound of claim 4, wherein the compound or salt thereof is about 98% pure.

7. 5. The compound of claim 4, wherein the compound or salt thereof is about 99% pure.

8. Formula II 【Chemistry 3】 or a salt thereof, wherein the compound or salt thereof has a purity of about 99.6%.

9. The compound is 【Chemistry 4】 or a salt thereof, wherein the compound or salt thereof is about 90% pure.

10. 10. The compound of claim 9, wherein the compound of formula IIa or a salt thereof is about 95% pure.

11. 10. The compound of claim 9, wherein the compound of formula IIa or a salt thereof is about 98% pure.

12. The compound is 【Chemistry 5】 or a salt thereof, wherein the compound or salt thereof is about 90% pure.

13. 13. The compound of claim 12, wherein the compound of formula IIb or a salt thereof is about 95% pure.

14. 13. The compound of claim 12, wherein the compound of formula IIb or a salt thereof is about 98% pure.

15. A pharma- ceutically acceptable excipient; 【Chemistry 6】 and at least one compound selected from the group consisting of aryl, ...

16. 16. The composition of claim 15, wherein at least one compound is a compound of formula Ia or a salt thereof, and the compound of formula Ia or a salt thereof is at least about 98% pure.

17. 16. The composition of claim 15, wherein at least one compound is a compound of formula Ib or a salt thereof, and the compound of formula Ib or a salt thereof is at least about 99% pure.

18. 16. The composition of claim 15 in the form of a capsule, tablet, or pill.

19. The compound of formula Ia according to claim 1 【Chemistry 7】 or a salt thereof, comprising: Formula VIa 【Chemistry 8】 with acryloyl chloride.

20. Deprotection in an acid system gives the compound of formula IX 【Chemistry 9】 20. The method of claim 19, further comprising preparing a compound of formula VIa from a compound of formula VIa.

21. The compound of formula Ib according to claim 4 【Chemistry 10】 or a salt thereof, comprising: Formula VIb 【Chemistry 11】 with acryloyl chloride.

22. Deprotection in an acid system gives the compound of formula VII 【Chemistry 12】 22. The method of claim 21, further comprising preparing a compound of formula VIb from a compound of formula VIb.

23. Deprotection using base gives the compound of formula VIII 【Chemistry 13】 23. The method of claim 22, further comprising preparing a compound of formula VII from a compound of formula VII.

24. Using triphenylphosphine and azodicarboxylate, a compound of formula IX 【Chemistry 14】 24. The method of claim 23, further comprising preparing a compound of formula VIII from a compound of formula VIII.

25. 25. The method of claim 24, wherein the azodicarboxylate is selected from diethyl azodicarboxylate and diisopropyl azodicarboxylate.

26. A compound as described herein substantially as hereinbefore.

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