Benzothiazole derivatives for the treatment of diseases

Abstract

The present disclosure relates to PDE4 inhibitor compounds, pharmaceutical compositions comprising the compounds, and methods of using such compounds to treat diseases.

Description

Related applications

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 565,592, filed March 15, 2024, the entire disclosure of which is incorporated herein by reference. Background Technology

[0002] Inhibition of PDE4 enzymes leads to the regulation of various inflammatory mechanisms through elevated levels of cyclic adenosine monophosphate (cAMP) in cells. The cAMP signaling system regulates many physiological and pathophysiological responses, including those in the cardiopulmonary, central nervous, cardiovascular, and immune systems, among which low intracellular levels can lead to many human diseases. One way to increase cellular cAMP concentration is by inhibiting PDEs to prevent cAMP hydrolysis. More than 21 PDE genes have been identified in the human genome, and these genes are classified into 11 families (PDE1 to PDE11) based on their sequence homology, substrate specificity, and inhibitor sensitivity. PDE4 enzymes are targets for the treatment of inflammatory diseases of the lungs, dermatology, and neurology. Several PDE4 inhibitors, including roflumilast, apremilast, and crisaborole, are approved and marketed as Dalsirep™, Otezla™, and Eucrisasa™ for the treatment of inflammatory lung diseases, psoriatic arthritis, and atopic dermatitis. However, these drugs have adverse effects, such as nausea, vomiting, and gastrointestinal effects. Selective inhibitors of PDE4 isoforms have the potential to reduce gastrointestinal side effects, including nausea and vomiting. The presence of the upstream conserved regions UCR1, UCR2, and CR3 distinguishes PDE4 from other PDE gene members, and selective interactions with these regions lead to selective inhibition of PDE4 isoforms.

[0003] Epilepsy is a disorder of the central nervous system in which abnormal electrical activity in the brain causes recurrent seizures, affecting consciousness, movement, and sensation. Many medications for seizures reduce the excitability of nerve cells in the brain, and they can also affect normal activity. For example, certain antiepileptic drugs (AEDs), such as levetiracetam and perampanel, are associated with undesirable behavioral side effects, including aggression and irritability. Other AEDs, such as topiramate, can cause undesirable cognitive impairments, including “brain fog” as well as memory and language complications. In pathological states, including epilepsy, increased PDE4 activity helps lower cAMP levels, which can exacerbate neuroinflammatory responses. Therefore, PDE4 inhibitors remain needed in approaches to treat diseases. For example, compounds that provide therapeutic benefits in central nervous system disorders such as epilepsy while minimizing undesirable behavioral side effects and complications are highly desirable. Summary of the Invention

[0004] In one aspect, this disclosure relates to compounds of formula I or pharmaceutically acceptable salts thereof.

[0005] (I)

[0006] in:

[0007] R 1 It is H, amino, or alkyl;

[0008] R 2 It is a heterocyclic group, cycloalkyl group, heteroaryl group, or aryl group; and

[0009] R 3 It is a heterocyclic group, cycloalkyl group, heteroaryl group, or aryl group.

[0010] In some respects, this disclosure relates to compounds of formula II, III, IIIa, IIIb or IV, or pharmaceutically acceptable salts thereof.

[0011] (II)

[0012] (III)

[0013] (IIIa)

[0014] (IIIb)

[0015] (IV)

[0016] Where R 1 R 2 R 3 R 4 R 5 X 1 X 2 and X 3 Each of these is as described in this article.

[0017] In some aspects, this disclosure provides a pharmaceutical composition comprising the compounds of this disclosure and pharmaceutically acceptable excipients.

[0018] In some aspects, this disclosure provides a method for treating a patient’s disease, the method comprising administering a compound or composition of this disclosure to a patient in need of such treatment. Attached Figure Description

[0019] Figure 1The graph shows the percentage increase of cAMP as a single mediator in HEK293 cells transfected with PDE4B1 (right panel) or in HEK293 cells with endogenous protein levels of PDE4A and PDE4D (left panel). Data are shown for the PDE4B selective inhibitor A33 and compound 1.

[0020] Figure 2A A graph showing the seizure frequency of all Scn1a+ / - mice exposed to compound 1 or the mediator, as measured by vEEG;

[0021] Figure 2B A graph showing the seizure frequency in Scn1a+ / - mice exposed to compound 1 or the agent and with severe phenotype* removed, as measured by vEEG. *Mice experiencing seizures ≥ Racine 4 during the first 48 hours of treatment were excluded from the seizure activity and lifespan assay. † Mixed-effects model (REML) two-way ANOVA, p = <0.0001, n = 6-8. BID = twice daily; Compound 1 administration: 40 mg / kg BID; Carrier administration: BID. BID = twice daily; vEEG = video electroencephalography;

[0022] Figure 3 The diagram shows the seizure activity of zebrafish with kv1.1 (kcna1- / -) and GABA_A1 (gabra1- / -) receptor knockout and PTZ-induced seizures induced by exposure to the mediator, compound 1, or other anti-seizure drugs (ASM). † Two-way ANOVA using Sidak multiple comparison test (p<0.0001): ** p<0.01; **** p<0.0001; ns = not significant. Compound 1 administration: 40 mg / kg BID; Carrier administration: BID. BID = twice daily; gabra1- / - = GABA_A receptor knockout α1 subunit; kcna1- / - = Kv1.1 potassium channel knockout; PTZ = pentylenetretazol, a model for inducing generalized epilepsy. Veh = carrier; WT = wild type;

[0023] Figure 4 A graph showing the percentage of protected wild-type (WT) mice exposed to either compound 1 or the mediator in the MES epilepsy model is presented. Compound 1 administration: 40 mg / kg BID; Mediator administration: BID. BID = twice daily;

[0024] Figure 5AA graph showing the Kaplan-Meier survival probabilities of mice exposed to compound 1 relative to those exposed to the mediator Scn1a+ / - epileptic mice is presented.

[0025] Figure 5B A graph showing the median survival of Scn1a+ / - epileptic mice after treatment cessation, relative to those exposed to the medium. † Unpaired t-test, p = 0.0081, n = 9–19; Compound 1 administration: 40 mg / kg BID; Carrier administration: BID. BID = twice daily; PND = postpartum days;

[0026] Figure 6A A graph showing the time spent near the target in the Barnes maze by WT and Scn1a+ / - epileptic mice exposed to compound 1 relative to WT mice exposed to the medium;

[0027] Figure 6B A graph showing the number of novel Y-maze arm entries in WT and Scn1a+ / - epileptic mice exposed to compound 1 relative to those exposed to the vehicle is presented. Compound 1 administration: 40 mg / kg BID; Vehicle administration: BID. BID = twice daily; Veh = vehicle; WT = wild-type;

[0028] Figure 7A A graph showing the number of embedded marbles in WT and Scn1a+ / - epileptic mice exposed to compound 1 relative to WT mice exposed to the medium;

[0029] Figure 7B A graph showing the distance traveled in an open field with a novel object in WT mice exposed to compound 1 versus those exposed to the medium is presented; and

[0030] Figure 7C A graph showing the time spent in the center of an open field with a novel object in WT mice exposed to compound 1 versus those exposed to the medium is shown. † One-way ANOVA, p = 0.0115, n = 3–11; ns = not significant; Compound 1 administration: 40 mg / kg BID; Carrier administration: BID. BID = twice daily; Veh = carrier; WT = wild type. Detailed Implementation

[0031] One object of this disclosure is to provide selective PDE4 inhibitors. The selective PDE4 inhibitors of this disclosure have found practical applications in the treatment of central nervous system disorders such as epilepsy, stroke, Alzheimer's disease, schizophrenia, depression, multiple sclerosis, traumatic brain injury, Huntington's disease, and other neurodevelopmental disorders. The selective PDE4 inhibitors of this disclosure that inhibit tumor growth and metastasis have found practical applications in the treatment and prevention of cancer. Such cancers may include esophageal cancer, brain cancer, pancreatic cancer, colon cancer, hematologic malignancies, lung cancer, prostate cancer, skin cancer, head and neck cancer, CNS cancer, gastric cancer, breast cancer, bladder cancer, or ovarian cancer. The selective PDE4 inhibitors of this disclosure can be used to improve cognition and therefore have found practical applications in the treatment of learning disabilities, memory loss, Fragile X syndrome, or other cognitive and emotional disorders. Selective PDE4 inhibitors can also be used to treat inflammatory diseases, including asthma, chronic obstructive pulmonary disease (COPD), psoriasis, atopic dermatitis, inflammatory bowel disease, and rheumatoid arthritis.

[0032] Before further describing this disclosure, it should be understood that this disclosure is not limited to the specific embodiments described, as these embodiments may, of course, vary. It should also be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting, as the scope of this disclosure will be defined only by the appended terms.

[0033] For the sake of brevity, all publications, including patents, cited in this specification are incorporated herein by reference. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. All patents, applications, published applications, and other publications mentioned herein are incorporated herein by reference in their entirety. If any definition set forth in this section is contrary to or otherwise inconsistent with the definition set forth in a patent, application, or other publication incorporated herein by reference, the definition set forth in this section shall prevail over the definition incorporated herein by reference.

[0034] As used herein and in the appended terms, the singular forms “a” and “the” include plural indicators unless the context clearly specifies otherwise. It should also be noted that the terms may be drafted to exclude any optional elements. Therefore, this statement is intended as a prior basis for the use of exclusive terms such as “uniquely” or “only” relating to the description of an element of the terms, or for the use of a “negative” restriction.

[0035] As used herein, the terms “including,” “containing,” and “comprise” are used in their open, non-restrictive sense.

[0036] To provide a more concise description, some quantitative expressions given herein are not limited by the term "about". It should be understood that, whether or not the term "about" is explicitly used, each quantity given herein means that it refers to an actual given value, and also to an approximation of such a given value that would be reasonably inferred based on ordinary techniques in the art, including equivalent and approximate values ​​resulting from the experimental and / or measurement conditions of such a given value. When yields are given as percentages, such yields refer to the mass of the entity for which the yield is given relative to the maximum amount of the same entity that could be obtained under specific stoichiometric conditions. Unless otherwise stated, concentrations given as percentages refer to mass ratios.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. While any methods and materials similar to or equivalent to those described herein may also be used in the practice or testing of this disclosure, preferred methods and materials are described hereafter. All publications referenced herein are incorporated by way of citation to disclose and describe methods and / or materials relating to the cited publications.

[0038] Unless otherwise stated, the methods and techniques of this embodiment are generally performed in accordance with conventional methods well known in the art and as described in the various general and more specific references cited and discussed throughout this specification. See, for example, Loudon, Organic Chemistry, 4th ed., New York: Oxford University Press, 2002, pp. 360-361, 1084-1085; Smith and March, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th ed., Wiley-Interscience, 2001.

[0039] The chemical names of the compounds described in this article have generally been derived using commercially available ACD / Name 2014 (ACD / Labs) or ChemBioDraw Ultra 13.0 (Perkin Elmer).

[0040] As used herein and in conjunction with the chemical structures depicting the various embodiments described herein, “*”, “**” and “” are used. "Each represents a covalent attachment point between a chemical group or chemical structure displaying that identifier and an adjacent chemical group or chemical structure. For example, in a hypothetical chemical structure AB, where A and B are connected by a covalent bond, in some embodiments, the portion of AB defined by group or chemical structure A can be..." , or This indicates that "-*", "-**" and "" are used in conjunction with the text. Each of the terms in the equation represents a bond with A and a covalent bond attachment point with B. Alternatively, in some embodiments, the portion of AB defined by the group or chemical structure B can be derived from... , or This indicates that "*", "-**", and "" are used to represent the characters in the text. Each of the symbols represents a bond attachment point to B and a covalent bond attachment point to A.

[0041] It should be understood that, for clarity, certain features of this disclosure described in the context of individual embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of this disclosure described in the context of individual embodiments may also be provided individually or in any suitable sub-combination. All combinations of embodiments relating to chemical groups represented by variables are specifically covered by this disclosure and disclosed herein as if each combination were disclosed individually and explicitly, to the extent that such combinations cover compounds as stable compounds (i.e., compounds whose biological activity can be isolated, characterized, and tested). Furthermore, all sub-combinations of chemical groups listed in embodiments describing such variables are also specifically covered by this disclosure and disclosed herein as if each such sub-combination of chemical groups were disclosed individually and explicitly herein.

[0042] definition

[0043] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by one of ordinary skill in the art. Generally, the nomenclature and techniques described herein in relation to chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry are those well-known and commonly used in the art.

[0044] Unless otherwise stated, the methods and techniques disclosed herein are generally practiced according to conventional methods well known in the art and as described in the various general and more specific references cited and discussed throughout this specification. See, for example, "Principles of Neural Science," McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intuitive Biostatistics," Oxford University Press, (1995); Lodish et al., "Molecular Cell Biology, 4th Edition," WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th Edition," WH Freeman & Co., New York (1999); and Gilbert et al., "Developmental Biology, 6th Edition," Sinauer Associates, Sunderland, MA (2000).

[0045] Unless otherwise defined herein, chemical terms used herein are used in accordance with conventional usage in the art, as illustrated by the McGraw-Hill Dictionary of Chemical Terms, Parker S., Ed., McGraw-Hill, San Francisco, California (1985).

[0046] All the foregoing references in this application, as well as any other publications, patents, and published patent applications, are specifically incorporated herein by reference. In case of conflict, this specification, including its specific definitions, shall prevail.

[0047] The term "pharmaceutical" is used herein to refer to chemical compounds (such as organic or inorganic compounds, mixtures of chemical compounds), biological macromolecules (such as nucleic acids, antibodies, including portions thereof, as well as humanized, chimeric, and human antibodies and monoclonal antibodies, proteins or portions thereof, such as peptides, lipids, carbohydrates), or extracts made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Pharmaceuticals include, for example, those with known structures and those with unknown structures. The ability of such pharmaceuticals to inhibit or promote AR degradation may make them suitable as "therapeutic agents" in the methods and compositions of this disclosure.

[0048] The terms “patient,” “subject,” or “individual” are used interchangeably and refer to human or non-human animals. These terms include mammals such as humans, primates, livestock (including bovines, swine, etc.), companion animals (e.g., canines, felines, etc.), and rodents (e.g., mice and rats).

[0049] "Treatment" of a condition or patient refers to measures taken to achieve a beneficial or desired outcome, including clinical outcomes. As used herein and as is well known in the art, "treatment" is a method for achieving a beneficial or desired outcome, including clinical outcomes. Beneficial or desired clinical outcomes may include, but are not limited to, relief or improvement of one or more symptoms or conditions, reduction of disease severity, stabilization of the disease state (i.e., non-deterioration), prevention of the spread of the disease, delay or slowing of disease progression, improvement or relief of the disease state, and remission (whether partial or complete), whether detectable or undetectable. "Treatment" may also mean extended survival compared to expected survival without treatment.

[0050] The term "prevention" is recognized in the art and is well known in the art when used in connection with conditions such as local recurrence (e.g., pain), diseases such as cancer, syndrome complexes such as heart failure, or any other medical condition. It includes administering a composition that, relative to a subject not receiving the composition, reduces the frequency of symptoms of the subject's medical condition or delays the onset of said symptoms. Thus, cancer prevention includes, for example, reducing the number of detectable cancerous growths in a group of patients receiving prophylactic treatment relative to an untreated control group, and / or delaying the appearance of detectable cancerous growths in a treated group, for example, by a statistically and / or clinically significant amount relative to an untreated control group.

[0051] The administration of a substance, compound, or agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, the compound or agent can be administered intravenously, intra-arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, through the eyes, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and percutaneously (by absorption, e.g., through a skin catheter). The compound or agent can also be suitably introduced via a rechargeable or biodegradable polymer device or other device such as a patch and pump or formulation, which provides an extended, slow, or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended time periods.

[0052] The appropriate method of administering a substance, compound, or agent to a subject will also depend on factors such as the subject's age and / or physical condition, and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability, and toxicity). In some embodiments, the compound or agent is administered orally, for example, by ingestion. In some embodiments, the orally administered compound or agent is in an extended-release or slow-release formulation, or is administered using a device for such slow-release or extended-release formulations.

[0053] As used herein, the phrase "combined administration" refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered agent remains effective in the body (e.g., both agents are effective simultaneously in the patient, which may include synergistic effects between the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially in the same formulation or in separate formulations. Thus, individuals receiving such treatment may benefit from the combined effects of different therapeutic agents.

[0054] The "therapeutic effective amount" or "therapeutic dose" of a drug or agent is the amount of drug or agent that will have the expected therapeutic effect when administered to a subject. A complete therapeutic effect does not necessarily occur with the administration of a single dose and may only occur after a series of doses. Therefore, the therapeutic effective amount may be administered in one or more doses. The precise effective amount required by the subject will depend on factors such as the subject's body size, health condition, and age, as well as the nature and extent of the condition being treated, such as cancer or MDS. Technicians can readily determine the effective amount for a given situation through routine laboratory procedures.

[0055] As used herein, the terms “optional” or “optionally” mean that an event or situation described below may or may not occur, and the description includes both cases where said event or situation occurs and cases where said event or situation does not occur. For example, “optionally substituted alkyl” means that the alkyl group may be substituted as well as cases where the alkyl group is not substituted.

[0056] It should be understood that those skilled in the art can select the substituents and substitution patterns on the compounds disclosed herein to produce chemically stable compounds that can be readily synthesized from readily available starting materials using techniques known in the art and the methods described below. If the substituent itself is substituted by more than one group, it should be understood that these multiple groups may be on the same carbon or different carbons, provided a stable structure is obtained.

[0057] As used herein, the term "optionally substituted" means replacing one to six hydrogen radicals in a given structure with a specified substituent, including but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclic, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2, or -CH2-OP(O)(O-alkyl)2. Preferably, "optionally substituted" means replacing one to four hydrogen radicals in a given structure with the aforementioned substituents. More preferably, one to three hydrogen radicals are replaced by the substituents described above. It should be understood that the substituents may be further substituted.

[0058] The term "acyl" is recognized in the art and refers to a group represented by the general formula hydrocarbon C(O)—, preferably a group represented by alkyl C(O)—.

[0059] The term "acylamino" is recognized in the art and refers to an amino group substituted with an acyl group, and may be represented, for example, by the alkyl group C(O)NH—.

[0060] The term "acyloxy group" is recognized in the art and refers to a group represented by the general formula hydrocarbon group C(O)O-, preferably a group represented by alkyl group C(O)O-.

[0061] The term "alkoxy group" refers to an alkyl group with an oxygen atom attached to it. Representative alkoxy groups include methoxy, ethoxy, propoxy, and tert-butoxy.

[0062] The term "alkoxyalkyl" refers to an alkyl group that has been substituted with an alkoxy group, and may be represented by the general formula alkyl-O-alkyl.

[0063] As used herein, the term "alkyl" refers to a saturated aliphatic group, including but not limited to C1-C1 groups. 10 Straight-chain alkyl groups or C1-C 10 Branched alkyl groups. Preferably, an "alkyl" group refers to a C1-C6 straight-chain alkyl group or a C1-C6 branched alkyl group. Most preferably, an "alkyl" group refers to a C1-C4 straight-chain alkyl group or a C1-C4 branched alkyl group. Examples of "alkyl" include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl, etc. Furthermore, the term "alkyl" as used throughout the specification, examples, and claims is intended to include both unsubstituted alkyl groups and substituted alkyl groups, the latter referring to an alkyl moiety having substituents on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.

[0064] Term "C" x-y "or "C x -C y "When used in conjunction with chemical motifs such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, it means to include a group containing x to y carbon atoms in the chain. C0 alkyl represents hydrogen, where the group is at the terminal position or, if internal, is a bond. For example, C1-6 alkyl groups contain one to six carbon atoms in the chain."

[0065] As used in this article, the term "amide" refers to the following groups:

[0066]

[0067] Where R 9 and R 10 Each independently represents a hydrogen or hydrocarbon group, or R 9 and R 10 Together with the N atoms to which they are attached, they form heterocycles with 4 to 8 atoms in the ring structure.

[0068] The terms "amine" and "amino" are recognized in the art and refer to both unsubstituted and substituted amines and their salts, for example, by the following terms:

[0069]

[0070] Where R 9 R 10 and R 10 Each independently represents a hydrogen or hydrocarbon group, or R 9 and R10 Together with the nitrogen atom to which they are attached, they form heterocycles with 4 to 8 atoms in the ring structure. Examples of amino groups include, but are not limited to, -NH2, -NH(alkyl), and -N(alkyl)2. 9 and R 10 Examples of heterocycles (e.g., cyclic amino groups) that form a ring structure with 4 to 8 atoms together with the N atoms to which they are attached include piperazine, pyrrolidine, piperidine, and morpholine.

[0071] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups, wherein each atom of the ring is a carbon atom. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more rings, wherein two or more carbons are shared by two adjacent rings, and at least one ring is aromatic; for example, other rings may be cycloalkyl, cycloalkenyl, cycloynyl, aryl, heteroaryl, and / or heterocyclic groups. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0072] The term "carbamate" is recognized in the art and refers to the following groups:

[0073]

[0074] Where R 9 and R 10 Independently represents a hydrogen or hydrocarbon group.

[0075] As used in this article, the term "carbocycloalkyl" refers to an alkyl group that has been substituted with a carbocyclo group.

[0076] The term "carbocyclic ring" includes 5-7 membered monocyclic rings and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocyclic ring may be selected from saturated rings, unsaturated rings, and aromatic rings. Carbocyclic rings include bicyclic molecules in which one, two, or three or more atoms are shared between two rings. The term "fused carbocyclic ring" refers to a bicyclic carbocyclic ring in which each ring shares two adjacent atoms with the other ring. Each ring of a fused carbocyclic ring may be selected from saturated rings, unsaturated rings, and aromatic rings. In an exemplary embodiment, an aromatic ring, such as a phenyl ring, may be fused with a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated bicyclic, unsaturated bicyclic, and aromatic bicyclic rings is included in the definition of a carbocyclic ring when valence permits. Exemplary "carbocyclic rings" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocyclic rings include decahydronaphthalene, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. The “carbocyclic ring” may be substituted at any one or more positions capable of carrying hydrogen atoms.

[0077] The term "carbonate" is recognized in the art and refers to the group —OCO2—.

[0078] As used in this article, the term "carboxyl group" refers to a group represented by the formula —CO2H.

[0079] As used herein, the term "cycloalkyl" refers to a 3- to 15-membered all-carbon monocyclic ring, including all-carbon 5-, 6-, or 6-, fused bicyclic rings, or polycyclic fused rings (a "fused" ring system means that each ring in the system shares an adjacent pair of carbon atoms with every other ring in the system), or a carbon ring fused to another group such as a heterocycle, such as a 5- or 6-membered cycloalkyl ring fused to a 5- to 7-membered heterocycle, wherein one or more rings may contain one or more double bonds, but the cycloalkyl ring does not contain a fully conjugated π-electron system. It should be understood that in some embodiments, the cycloalkyl ring may advantageously have a limited size, such as C3-C4. 13 C3-C9, C3-C6, and C4-C6. The cycloalkyl group may be unsubstituted or substituted as described with respect to alkyl groups or as described in the various embodiments provided herein. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, adamantyl, norbornyl, norbornenyl, etc. Exemplary examples of cycloalkyl groups shown in the illustrations include the following entities in the form of appropriately bonded portions:

[0080]

[0081]

[0082] ,and .

[0083] As used in this article, "cyano" refers to the -CN group.

[0084] As used in this article, the term "ester" refers to the group —C(O)OR 8 , where R 8 It represents a hydrocarbon group.

[0085] As used in this article, the term "ketone" refers to the group —C(O)R 7 , where R 7 This indicates a hydrocarbon group (e.g., alkyl, aryl, heteroaryl). An exemplary example of a ketone group is a methyl ketone (—C(O)CH3).

[0086] As used herein, the term "ether" refers to a hydrocarbon group that is attached to another hydrocarbon group by oxygen. Therefore, the ether substituent of a hydrocarbon group may be alkyl-O-. Ethers may be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocyclic-O-heterocycles and aryl-O-heterocycles. Ethers include "alkoxyalkyl" groups, which may be represented by the general formula alkyl-O-alkyl.

[0087] As used herein, the terms “halo” and “halogen” refer to halogens and include chlorine, fluorine, bromine, and iodine.

[0088] As used herein, the terms “hetaralkyl” and “heteroaralkyl” refer to alkyl groups that have been substituted with heteroaryl groups.

[0089] The terms "heteroaryl" and "hetaryl" encompass substituted or unsubstituted aromatic monocyclic structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structure includes at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic systems having two or more rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is heteroaromatic; for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic groups. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Exemplary heteroaryl groups include, for example, pyrazole, imidazole, oxazole, pyridine, pyridinone, pyrimidine, and pyridazine.

[0090] As used herein, the term "heteroatom" refers to an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0091] As used in this article, the term "heterocyclic alkyl" refers to an alkyl group that has been substituted with a heterocyclic group.

[0092] The terms "heterocyclic group," "heterocyclic," and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structure includes at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclic group" and "heterocyclic" also include polycyclic systems having two or more rings, wherein two or more carbons are shared by two adjacent rings, and wherein at least one ring is a heterocyclic ring; for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic groups. Heterocyclic groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.

[0093] As used herein, the term "hydrocarbon group" refers to a group bonded by carbon atoms, which does not have ═O or ═S substituents and typically has at least one carbon-hydrogen bond and a predominant carbon backbone, but may optionally include heteroatoms. Therefore, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbon groups, but substituents such as acetyl (which has an ═O substituent on the linking carbon) and ethoxy (which is linked by oxygen rather than carbon) are not considered hydrocarbon groups. Hydrocarbon groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, ynyl, and combinations thereof.

[0094] As used in this article, the term "hydroxyalkyl" refers to an alkyl group that has been replaced by a hydroxyl group.

[0095] The term "lower" when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy means including groups in which ten or fewer atoms, preferably six or fewer atoms, are present in the substituents. For example, "lower alkyl" refers to an alkyl group containing ten or fewer carbon atoms, preferably six or fewer carbon atoms. In some embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents as defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as when hydroxyalkyl and aralkyl are described (in which case, for example, when counting the carbon atoms in the alkyl substituents, the atoms in the aryl group are not counted).

[0096] The terms "polycyclic," "polycyclic," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic) in which two or more atoms are shared by two adjacent rings; for example, the rings are "fused rings." Each ring in a polycyclic ring may be substituted or unsubstituted. In some embodiments, each ring of the polycyclic ring contains 3 to 10 atoms, preferably 5 to 7 atoms.

[0097] The term "sulfate" is recognized in the art and refers to the group —OSO3H or its pharmaceutically acceptable salt.

[0098] The term "sulfonamide" is recognized in the art and refers to a group represented by the following general formula:

[0099]

[0100] Where R 9 and R 10 Independently represents either hydrogen or hydrocarbon groups.

[0101] The term "sulfoxide" is recognized in the art and refers to the group —S(O)—.

[0102] The term "sulfonate" is recognized in the art and refers to the group SO3H or its pharmaceutically acceptable salt.

[0103] The term "sulfone" is recognized in the art and refers to the group —S(O)2-R 9 , where R 9 This indicates a hydrocarbon group (e.g., alkyl, aryl, heteroaryl). An exemplary example of a sulfone group is methyl sulfone (—S(O)2CH3).

[0104] The term "substituted" refers to a portion having a substituent having a hydrogen atom on one or more carbons of the main chain. It should be understood that "substituted" or "replaced by" includes the implicit condition that such substitution meets the permissible valence of the substituted atom and the substituent, and that said substitution produces a stable compound, for example, which does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is contemplated to include all permissible substituents of an organic compound. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. For a suitable organic compound, permissible substituents may be one or more, and may be the same or different. For the purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of the organic compound described herein that satisfy the valence of the heteroatom. Substituents may include any substituents described herein, such as halogens, hydroxyl groups, carbonyl groups (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl groups (such as thioesters, thioacetic acids, or thiocarbamates), alkoxy groups, phosphoryl groups, phosphate esters, phosphonates, hypophosphonates, amino groups, amide groups, amidine groups, imine groups, cyano groups, nitro groups, azide groups, mercapto groups, alkylthio groups, sulfate esters, sulfonates, aminosulfonyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, aralkyl groups, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that, where appropriate, the portion substituted on the hydrocarbon chain may itself be substituted.

[0105] As used in this article, the term "regulation" includes both inhibiting or blocking functions or activities (such as cell proliferation) and enhancing functions or activities.

[0106] The phrase “pharmaceutically acceptable” is recognized in the art. In some embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms that, to the extent of reasonable medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0107] "Pharmaceutically acceptable salt" or "salt" as used in this article refers to an acid addition salt or base addition salt that is suitable for or compatible with the treatment of the patient.

[0108] As used herein, the term "pharmaceutically acceptable acid addition salt" means any non-toxic organic or inorganic salt of any base compound represented by Formula I. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include monocarboxylic acids, dicarboxylic acids, and tricarboxylic acids, such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, and salicylic acid, as well as sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Monoacid salts or diacid salts can be formed, and such salts may exist in hydrated, solvated, or substantially anhydrous forms. Generally, acid addition salts of compounds of Formula I are more soluble in water and a variety of hydrophilic organic solvents and typically exhibit a higher melting point compared to their free base form. The selection of suitable salts will be known to those skilled in the art. Other non-pharmaceutical acceptable salts, such as oxalates, may be used, for example, for compounds of formula I for laboratory use, or for subsequent conversion into pharmaceutically acceptable acid addition salts.

[0109] As used herein, the term "pharmaceutically acceptable base addition salt" refers to any non-toxic organic or inorganic base addition salt of any acid compound or any intermediate thereof represented by Formula I. Exemplary inorganic bases forming suitable salts include hydroxides of lithium, sodium, potassium, calcium, magnesium, or barium. Exemplary organic bases forming suitable salts include aliphatic, alicyclic, or aromatic organic amines, such as methylamine, trimethylamine, and methylpyridine or ammonia. The selection of suitable salts will be known to those skilled in the art.

[0110] Many compounds useful in the methods and compositions of this disclosure have at least one stereocenter in their structure. The stereocenter may be present in an R or S configuration, the R and S notations being consistent with the rules described in Pure Appl. Chem. (1976), 45, 11-30. This disclosure considers all stereoisomers, such as enantiomers and diastereomers (including all possible mixtures of stereoisomers) of compounds, salts, prodrugs, or mixtures thereof. See, for example, WO 01 / 062726.

[0111] Furthermore, some compounds containing alkenyl groups may exist as Z (zusammen) or E (entgegen) isomers. In each case, this disclosure includes both mixtures and separate, individual isomers.

[0112] Some compounds may also exist as tautomers. Although not explicitly indicated in the formulas described herein, such forms are intended to be included within the scope of this disclosure. For example, compounds having tautomer groups such as pyridone / hydroxypyridinium groups may be represented by any tautomer form.

[0113] A “prodrug” or “pharmaceutically acceptable prodrug” is a compound that, upon administration, is metabolized in a host body, for example, by hydrolysis or oxidation, to form a compound of this disclosure (e.g., a compound of formula I). ​​Typical examples of prodrugs include compounds having a biologically unstable or cleavable (protective) group on the functional portion of an active compound. Prodrugs include compounds that can be oxidized, reduced, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydrolysis, alkylation, dealkylation, acylation, deacylation, phosphorylation, or dephosphorylation to produce an active compound. Examples of prodrugs using esters or aminophosphates as biologically unstable or cleavable (protective) groups are disclosed in U.S. Patent Nos. 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of this disclosure are metabolized to produce compounds of formula I. This disclosure includes prodrugs of the compounds described herein within its scope. The routine procedures for selecting and preparing suitable prodrugs are described, for example, in “Design of Prodrugs”, Ed. H. Bundgaard, Elsevier, 1985.

[0114] As used in this article, the phrase “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or medium that can be used to formulate a drug for medical or therapeutic purposes, such as a liquid or solid filter, diluent, excipient, solvent, or encapsulating material.

[0115] As used herein, the terms “logarithm of solubility,” “LogS,” or “logS” are used in the art to quantify the water solubility of a compound. The water solubility of a compound significantly affects its absorption and distribution characteristics. Low solubility is typically accompanied by poor absorption. The LogS value is the dimensionless logarithm (base 10) of solubility measured in moles per liter.

[0116] As used herein, the term "PDE4 inhibitor" refers to a compound that prevents, reverses, slows down, or inhibits the activity of PDE4 or PDE4 isoforms.

[0117] In the treatment methods according to this disclosure, "effective amount" means an amount or dose sufficient to generally provide the desired therapeutic benefit in a subject requiring such treatment. The effective amount or dose of the compounds of this disclosure may be determined by conventional methods such as modeling, dose escalation, or clinical trials, taking into account conventional factors such as the manner or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the infection, the subject's health status, condition and weight, and the judgment of the treating physician. Exemplary doses range from about 0.1 mg to 1 g per day, or from about 1 mg to 50 mg per day, or from about 50 to 250 mg per day, or from about 250 mg to 1 g per day. The total dose may be given in single-dose units or fractional-dose units (e.g., BID, TID, QID).

[0118] Representative implementation plan

[0119] In some embodiments, this disclosure relates to compounds of formula I or pharmaceutically acceptable salts thereof.

[0120] (I),

[0121] Where R 1 R 2 and R 3 As described in this article.

[0122] In some embodiments, this disclosure relates to compounds of formula II or pharmaceutically acceptable salts thereof.

[0123] (II)

[0124] Where R 2 and R 3 As described in this article.

[0125] In some embodiments, this disclosure relates to compounds of formula III or pharmaceutically acceptable salts thereof.

[0126] (III)

[0127] Where R 1 R 3 X 1 X 2 and X 3 As described in this article.

[0128] In some embodiments, this disclosure relates to compounds of formula IIIa or pharmaceutically acceptable salts thereof.

[0129] (IIIa)

[0130] Where R1 R 3 R 5 X 1 and X 3 As described in this article.

[0131] In some embodiments, this disclosure relates to compounds of formula IIIb or pharmaceutically acceptable salts thereof.

[0132] (IIIb)

[0133] Where R 1 R 4 and R 5 As described in this article.

[0134] In some embodiments, this disclosure relates to compounds of formula IV or pharmaceutically acceptable salts thereof.

[0135] (IV)

[0136] Where R 4 As described in this article.

[0137] In some implementation schemes, R 1 It is H, amino, or alkyl. In some embodiments, R 1 It is H, NH2, or a C1-C6 alkyl group. In some embodiments, R 1 It is H or an amino group. In some embodiments, R 1 It is H or NH2. In some implementations, R 1 It is H. In some preferred embodiments, R 1 It is NH2.

[0138] In some implementation schemes, R 2 It is a heterocyclic, cycloalkyl, heteroaryl, or aryl group. In some embodiments, R 2 It is a 3- to 8-membered heterocyclic group, a C3-C8 cycloalkyl group, a 3- to 8-membered heteroaryl group, or a C6-C group. 10 Aryl. In some implementations, R 2 It is a heteroaryl or aryl group. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl or C6-C 10 Aryl. In some implementations, R 2 It is a 5- to 6-membered heteroaryl or C6-C 10 Aryl.

[0139] In some implementation schemes, R 2 It is a heteroaryl group. In some implementations, R 2 It is a 3- to 8-membered heteroaryl group. In some implementation schemes, R2 It is a 5- or 6-membered heteroaryl group. In some preferred embodiments, R 2 It is a 6-membered heteroaryl group.

[0140] In some implementation schemes, R 2 It is pyridyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl. In some embodiments, R 2 It is pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl. In some preferred embodiments, R 2 It is pyridyl (e.g., 3-pyridyl).

[0141] In some implementation schemes, R 2 Optionally substituted with an amino group (e.g., NH2, morpholino, or piperazine) or an alkyl group (e.g., C1-C6 alkyl). In some embodiments, R 2 Optionally substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2 Optionally substituted with NH2 or methyl. In some embodiments, R 2 Optionally replaced by NH2. In some implementations, R 2 Optionally substituted with a methyl group. In some embodiments, R 2 Optionally substituted with NH2, morpholino, piperazine, or C1-C6 alkyl. In some embodiments, R 2 Optionally substituted with a morpholino group. In some embodiments, R 2 Optionally substituted with piperazine group.

[0142] In some implementation schemes, R 2 It is substituted with an amino or alkyl group. In some embodiments, R 2 It is substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2 It is replaced by NH2 or methyl. In some embodiments, R 2 Replaced by NH2. In some implementations, R 2 It is replaced by C1-C6 alkyl groups (e.g., methyl).

[0143] In some implementation schemes, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with an amino or alkyl group. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with NH2, morpholino, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2It is a 3- to 8-membered heteroaryl group optionally substituted with NH2, morpholino, piperazine, or methyl. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with morpholino or piperazine. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with NH2. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with a morpholino group. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group optionally substituted with a piperazine group. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group that is optionally substituted with a methyl group.

[0144] In some implementation schemes, R 2 It is a 3- to 8-membered heteroaryl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group substituted with NH2 or methyl. In some embodiments, R 2 It is a 3- to 8-membered heteroaryl group substituted with NH2. In some embodiments, R 2 It consists of 3- to 8-membered heteroaryl groups substituted with methyl groups.

[0145] In some implementation schemes, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with an amino or alkyl group. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with NH2, morpholino, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with NH2, morpholino, piperazine, or methyl. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with morpholino or piperazine. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with NH2. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with a morpholino group. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group optionally substituted with a piperazine group. In some embodiments, R2 It is a 5- to 6-membered heteroaryl group that is optionally substituted with a methyl group.

[0146] In some implementation schemes, R 2 It is a 5- to 6-membered heteroaryl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with NH2, morpholino, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with NH2, morpholino, piperazine, or methyl. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with NH2 or methyl. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with morpholino or piperazine. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with NH2. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with a morpholino group. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with a piperazine group. In some embodiments, R 2 It is a 5- to 6-membered heteroaryl group substituted with methyl.

[0147] In some implementation schemes, R 2 It is an unsubstituted 3- to 8-membered heteroaryl group. In some embodiments, R 2 It is an unsubstituted 5- to 6-membered heteroaryl group. In some preferred embodiments, R 2 It is an unsubstituted 6-membered heteroaryl group.

[0148] In some implementation schemes, R 2 It is optionally substituted with an amino or alkyl group, and can be a pyridinyl, pyridone, pyrimidinyl, pyridazinyl, imidazoleyl, or oxazolyl group. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with NH2, morpholinyl, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is optionally substituted with NH2 or C1-C6 alkyl groups, namely pyridinyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazole, or oxazolyl group optionally substituted with NH2, morpholinyl, piperazine, or methyl. In some embodiments, R 2 It is a pyridinyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with NH2 or methyl. In some embodiments, R 2It is a pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with morpholinyl or piperazineyl. In some embodiments, R 2 It is optionally substituted with NH2, and consists of a pyridinyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with a morpholinyl group. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with a piperazine group. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with a methyl group.

[0149] In some implementation schemes, R 2 It is optionally substituted with an amino or alkyl group, and is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group. In some embodiments, R 2 It is optionally substituted with NH2 or C1-C6 alkyl groups, namely pyridinyl, pyrimidinyl, pyrazinyl, pyrazolyl, imidazoleyl, or oxazolyl. In some embodiments, R 2 It is optionally substituted with NH2 or methyl groups of pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl groups. In some embodiments, R 2 It is optionally substituted with NH2, and consists of a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group optionally substituted with a methyl group.

[0150] In some implementation schemes, R 2 It is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group substituted with NH2 or C1-C6 alkyl. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group substituted with NH2 or a methyl group. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group substituted with NH2. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl group substituted with a methyl group.

[0151] In some implementation schemes, R 2It is an unsubstituted pyridyl group, an unsubstituted pyridone group, an unsubstituted pyrimidinyl group, an unsubstituted pyridazinyl group, an unsubstituted imidazoleyl group, or an unsubstituted oxazolyl group. In some embodiments, R 2 It is an unsubstituted pyridyl, unsubstituted pyrimidinyl, unsubstituted pyridazinyl, unsubstituted pyrazolyl, unsubstituted imidazolyl, or unsubstituted oxazolyl.

[0152] In some implementation schemes, R 2 It is optionally substituted with an amino or alkyl group, such as a pyridinyl, pyridone, pyrimidinyl, or pyridazinyl group. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, or pyridazinyl group optionally substituted with NH2, morpholinyl, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is optionally substituted with NH2 or C1-C6 alkyl groups, namely pyridinyl, pyridone, pyrimidinyl, or pyridazinyl. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, or pyridazinyl group optionally substituted with a morpholinyl or piperazine group. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, or pyridazinyl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is optionally a pyridinyl, pyridone, pyrimidinyl, or pyridazinyl group substituted with NH2. In some embodiments, R 2 It is a pyridyl, pyridone, pyrimidinyl, or pyridazinyl group optionally substituted with a morpholinyl group. In some embodiments, R 2 It is a pyridinyl, pyridone, pyrimidinyl, or pyridazinyl group optionally substituted with a piperazine group.

[0153] In some implementation schemes, R 2 It is optionally substituted with an amino or alkyl group, and is a pyridinyl, pyrimidinyl, or pyridazinyl group. In some embodiments, R 2 It is optionally pyridinyl, pyrimidinyl, or pyridazinyl substituted with NH2 or C1-C6 alkyl. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, or pyridazinyl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, or pyridazinyl group that is optionally substituted with NH2.

[0154] In some implementation schemes, R 2 It is a pyridinyl, pyrimidinyl, or pyridazinyl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, or pyridazinyl group substituted with NH2 or a C1-C6 alkyl group. In some embodiments, R 2 It is a pyridinyl, pyrimidinyl, or pyridazinyl group substituted with NH2 or a methyl group. In some embodiments, R 2It is a pyridinyl, pyrimidinyl, or pyridazinyl group substituted with NH2.

[0155] In some implementation schemes, R 2 It is an unsubstituted pyridyl group, an unsubstituted pyridone group, an unsubstituted pyrimidinyl group, or an unsubstituted pyridazinyl group. In some embodiments, R 2 It is an unsubstituted pyridinyl group, or an unsubstituted pyridinone. In some embodiments, R 2 It is an unsubstituted pyridyl, unsubstituted pyrimidinyl, or unsubstituted pyridazinyl group. In some embodiments, R 2 It is an unsubstituted pyridinyl group (e.g., unsubstituted 3-pyridinyl). In some embodiments, R 2 It is an unsubstituted pyrimidine group. In some embodiments, R 2 It is an unsubstituted pyridazinyl group. In some embodiments, R 2 It is an unsubstituted pyridinone.

[0156] In some implementation schemes, R 2 It is optionally substituted with an amino or alkyl group, such as a pyrazolyl, imidazole, or oxazolyl group. In some embodiments, R 2 It is optionally a pyrazolyl, imidazole, or oxazolyl group substituted with NH2 or a C1-C6 alkyl group. In some embodiments, R 2 It is a pyrazolyl, imidazole, or oxazolyl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a pyrazolyl, imidazole, or oxazolyl group that is optionally substituted with NH2.

[0157] In some implementation schemes, R 2 It is a pyrazolyl, imidazole, or oxazolyl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyrazolyl, imidazole, or oxazolyl group substituted with NH2 or a C1-C6 alkyl group. In some embodiments, R 2 It is a pyrazolyl, imidazole, or oxazolyl group substituted with NH2 or methyl. In some embodiments, R 2 It is a pyrazolyl group, imidazole group, or oxazolyl group that is replaced by NH2.

[0158] In some implementation schemes, R 2 It is an unsubstituted pyrazolyl, imidazole, or oxazolyl group. In some embodiments, R 2 It is an unsubstituted pyrazolyl group. In some embodiments, R 2 It is an unsubstituted imidazole group. In some embodiments, R 2 It is an unsubstituted oxazolyl group.

[0159] In some implementation schemes, R 2It is a pyridyl group optionally substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyridyl group optionally substituted with NH2, morpholino, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is a pyridinyl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a pyridyl group optionally substituted with NH2, morpholino, or piperazine. In some embodiments, R 2 It is a pyridinyl group substituted with NH2. In some embodiments, R 2 It is a pyridyl group optionally substituted with a morpholino group. In some embodiments, R 2 It is a pyridyl group that is optionally substituted with a piperazine group.

[0160] In some implementation schemes, R 2 It is a pyridyl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyridyl group substituted with NH2, morpholino, piperazine, or C1-C6 alkyl. In some embodiments, R 2 It is a pyridyl group substituted with NH2 or methyl. In some embodiments, R 2 It is a pyridyl group substituted with NH2, morpholino, or piperazine. In some embodiments, R 2 It is a pyridinyl group substituted with NH2. In some embodiments, R 2 It is a pyridyl group substituted with a morpholino group. In some embodiments, R 2 It is a pyridyl group substituted with a piperazine group.

[0161] In some implementation schemes, R 2 It is a pyrimidinyl group optionally substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyrimidinyl group optionally substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2 It is a pyrimidinyl group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a pyrimidine group that is optionally substituted with NH2.

[0162] In some implementation schemes, R 2 It is a pyrimidinyl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyrimidinyl group substituted with NH2 or C1-C6 alkyl. In some embodiments, R 2 It is a pyrimidinyl group substituted with NH2 or methyl. In some embodiments, R 2 It is a pyrimidine group substituted with NH2.

[0163] In some implementation schemes, R 2It is an imidazole group optionally substituted with an amino or alkyl group. In some embodiments, R 2 It is an imidazole group optionally substituted with NH2 or C1-C6 alkyl. In some embodiments, R 2 It is an imidazole group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is an imidazolyl group that is optionally substituted with a methyl group.

[0164] In some implementation schemes, R 2 It is an imidazole group substituted with an amino or alkyl group. In some embodiments, R 2 It is an imidazole group substituted with NH2 or C1-C6 alkyl. In some embodiments, R 2 It is an imidazole group substituted with NH2 or methyl. In some embodiments, R 2 It is an imidazole group substituted with a methyl group.

[0165] In some implementation schemes, R 2 It is a pyrazolyl group optionally substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyrazolyl group optionally substituted with NH2 or C1-C6 alkyl groups. In some embodiments, R 2 It is a pyrazol group optionally substituted with NH2 or methyl. In some embodiments, R 2 It is a pyrazolyl group that is optionally substituted with a methyl group.

[0166] In some implementation schemes, R 2 It is a pyrazolyl group substituted with an amino or alkyl group. In some embodiments, R 2 It is a pyrazolyl group substituted with NH2 or C1-C6 alkyl. In some embodiments, R 2 It is a pyrazol group substituted with NH2 or methyl. In some embodiments, R 2 It is a pyrazol group substituted with a methyl group.

[0167] In some implementation schemes, R 2 Choose from the following groups:

[0168] , , , , , , , , , and .

[0169] In some implementation schemes, R 2 Choose from the following groups:

[0170] , , , , , and .

[0171] In some implementation schemes, R 2 Choose from the following groups:

[0172] , , and .

[0173] In some implementation schemes, R 2 Choose from the following groups:

[0174] , , and .

[0175] In some implementation schemes, R 2 Choose from the following groups:

[0176] , , , , , , and .

[0177] In some implementation schemes, R 2 Choose from the following groups:

[0178] , and .

[0179] In some implementation schemes, R 3 It is a heterocyclic, cycloalkyl, heteroaryl, or aryl group. In some embodiments, R 3 It is a 3- to 8-membered heterocyclic group, a C3-C8 cycloalkyl group, a 3- to 8-membered heteroaryl group, or a C6-C group. 10 Aryl. In some implementations, R 3 It is a heterocyclic, heteroaryl, or aryl group. In some embodiments, R 3 It is a 3- to 8-membered heterocyclic group, a 3- to 8-membered heteroaryl group, or a C6-C group. 10 Aryl. In some implementations, R 3 It is a heteroaryl or aryl group. In some embodiments, R 3It is a 3- to 8-membered heteroaryl or C6-C 10 Aryl. In some preferred embodiments, R 3 It is an aryl group (e.g., phenyl).

[0180] In some implementation schemes, R 3 It is a heterocyclic, cycloalkyl, heteroaryl, or aryl group, wherein each hydrogen atom in the heterocyclic, cycloalkyl, heteroaryl, or aryl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a heterocyclic, heteroaryl, or aryl group, wherein each hydrogen atom in the heterocyclic, heteroaryl, or aryl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group.

[0181] In some implementation schemes, R 3 It is a heterocyclic group. In some implementations, R 3 It is a 3- to 8-membered heterocyclic group. In some implementations, R 3 It is a 3- to 8-membered heterocyclic group. In some implementations, R 3 It is a 3- to 8-membered heterocyclic group, wherein each hydrogen atom in the 3- to 8-membered heterocyclic group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a 6-membered heterocyclic group. In some implementations, R 3 It is a 6-membered heterocyclic group, wherein each hydrogen atom in the 6-membered heterocyclic group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl. In some embodiments, R 3 It is tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl, wherein each hydrogen atom in tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl, wherein each hydrogen atom in tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl is optionally substituted with an alkyl group. In some embodiments, R 3 It is tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl, wherein each hydrogen atom in tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl is optionally substituted with a C1-C6 alkyl group. In some embodiments, R 3 It is tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl, wherein each hydrogen atom in the cyclohexyl group is optionally substituted with a methyl group. In some embodiments, R 3 It is N-methylpiperidinyl or N-methyltetrahydropyridinyl. In some embodiments, R 3It is an unsubstituted tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl.

[0182] In some implementation schemes, R 3 It is a cycloalkyl group. In some embodiments, R 3 It is C6-C 10 Cycloalkyl. In some embodiments, R 3 It is cyclohexyl. In some implementations, R 3 It is a cyclohexyl group, wherein each hydrogen atom in the cyclohexyl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a cyclohexyl group, wherein one or two hydrogen atoms of the cyclohexyl group are optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a cyclohexyl group, wherein each hydrogen atom in the cyclohexyl group is optionally replaced by a halogen. In some embodiments, R 3 It is a cyclohexyl group, wherein one or both hydrogen atoms of the cyclohexyl group are optionally substituted with halogens. In some embodiments, R 3 It is a cyclohexyl group, wherein each hydrogen atom in the cyclohexyl group is optionally replaced by chlorine, bromine, iodine, or fluorine. In some embodiments, R 3 It is a cyclohexyl group, wherein each hydrogen atom in the cyclohexyl group is optionally replaced by fluorine. In some embodiments, R 3 It is 4,4-difluorocyclohexyl. In some embodiments, R 3 It is an unsubstituted cyclohexyl group.

[0183] In some implementation schemes, R 3 It is a heteroaryl group. In some implementations, R 3 It is a 3- to 8-membered heteroaryl group. In some implementation schemes, R 3 It is a 5- or 6-membered heteroaryl group. In some implementations, R 3 It is a 5- or 6-membered heteroaryl group, wherein each hydrogen atom in the 5- or 6-membered heteroaryl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is thiophene-based. In some implementations, R... 3 It is a thiophene group, wherein each hydrogen atom in the thiophene group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a thiophene group, wherein each hydrogen atom in the thiophene group is optionally replaced by a halogen. In some embodiments, R 3 It is a thiophene group, wherein each hydrogen atom in the thiophene group is optionally replaced by chlorine, bromine, iodine, or fluorine. In some embodiments, R 3 It is a thiophene group, wherein each hydrogen atom in the thiophene group is optionally replaced by chlorine. In some embodiments, R3 It is 2-chloro-thiophene. In some embodiments, R 3 It is an unsubstituted thiophene group.

[0184] In some implementation schemes, R 3 It is aryl. In some implementations, R 3 It is C6-C 10 Aryl. In some implementations, R 3 It is C6-C 10 Aryl, wherein C6-C 10 Each hydrogen atom in the aryl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is phenyl. In some embodiments, R 3 It is a phenyl group, wherein each hydrogen atom in the phenyl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a phenyl group, wherein one hydrogen atom of the phenyl group is optionally substituted with a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is a phenyl group, wherein the hydrogen atom at the 3-position of the phenyl group is substituted with a halogen (e.g., chlorine), cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 3 It is an unsubstituted phenyl group. In some preferred embodiments, R 3 It is a phenyl group, wherein the hydrogen atom at the 3-position of the phenyl group is substituted with a halogen (e.g., chlorine) or a nitro group. In some preferred embodiments, R 3 It is 3-chlorophenyl or 3-nitrophenyl. In some preferred embodiments, R 3 It is 3-chlorophenyl.

[0185] In some implementation schemes, R 3 Choose from the following groups:

[0186] , , , , , , , , , , and .

[0187] In some implementation schemes, R 3 Choose from the following groups:

[0188] , , , and .

[0189] In some implementation schemes, R 3 Choose from the following groups:

[0190] , , , , and .

[0191] In some implementation schemes, R 4 It is a halogen, cyano, sulfone, nitro, ketone, or alkyl group. In some embodiments, R 4 It is a halogen, cyano, sulfone, nitro, or ketone. In some embodiments, R 4 It is chlorine, CN, SO2Me, NO2, or C(O)Me. In some preferred embodiments, R 4 It is a halogen (e.g., chlorine).

[0192] In some implementation schemes, X 1 and X 3 Each of them is independently CH or N. In some implementations, X 1 It is CH or N. In some implementations, X 3 It is CH or N. In some implementations, X 1 It is CH. In some implementations, X 1 It is N. In some implementations, X 3 It is CH. In some implementations, X 3 It is N. In some implementations, when X 1 When it is N, X 2 It is CH. In some implementations, X 3 It is N.

[0193] In some implementation schemes, X 2 It is CR 5 In some implementations, X 2 It is CH or C-NH2. In some implementations, X 2 It is CH. In some implementations, X 2 It is C-NH2. In some implementations, when X 1 When it is N, X 2 It is CH. In some implementations, when X 3 When it is N, X 1 It is CH. In some implementations, when X 1 When it is N, X 2 and X 3It is CH. In some implementations, X 1 and X 3 Each of them is CH, and X 2 It is CR 5 In some implementations, X 1 and X 3 Each of them is CH, and X 2 It is CH or C-NH2. In some implementations, X 1 and X 3 Each of them is CH, and X 2 It is C-NH2. In some implementations, X 1 X 2 and X 3 Each of them is CH.

[0194] In some implementation schemes, R 5 It is H or an amino group (e.g., -NH2, morpholino, or piperazine). In some preferred embodiments, R 5 It is H. In some implementations, R 5 It is an amino group.

[0195] In some implementations, when R 1 When it is H, R 2 It is pyridyl. In some embodiments, when R... 3 When it is a heterocyclic or cycloalkyl group, R 2 It is pyridyl. In some embodiments, when R... 1 When it is H, R 3 It is phenyl or thiophene.

[0196] In some implementation schemes, R 1 It is an amino group (e.g., NH2), and R 2 It is a heteroaryl group (e.g., pyridyl, such as 3-pyridyl). In some embodiments, R 1 It is an amino group (e.g., NH2), and R 3 It is an aryl group (e.g., phenyl, such as 3-chlorophenyl). In some embodiments, R 2 It is a heteroaryl group (e.g., pyridyl, such as 3-pyridyl), and R 3 It is an aryl group (e.g., phenyl, such as 3-chlorophenyl).

[0197] In some embodiments, this disclosure relates to compounds of formula I or pharmaceutically acceptable salts thereof, wherein R 1 It is an amino group (e.g., NH2), R 2 It is a heteroaryl group (e.g., pyridyl, such as 3-pyridyl), and R 3 It is an aryl group (e.g., phenyl, such as 3-chlorophenyl).

[0198] In some implementations, the compounds are selected from the group consisting of:

[0199] , , ,

[0200] , , , , , , , , , , , , , , , , , , , , ,

[0201] , , , , , , and ,

[0202] Or its pharmaceutically acceptable salt.

[0203] In some implementations, the compounds are selected from the group consisting of:

[0204] , , , , , , , , , , , , , , , , , , , , , , and ,

[0205] Or its pharmaceutically acceptable salt.

[0206] In some implementations, the compounds are selected from the group consisting of:

[0207] , , , , , , and ,

[0208] Or its pharmaceutically acceptable salt.

[0209] In some implementations, the compounds are selected from the group consisting of:

[0210] , , ,

[0211] , , , , and ,

[0212] Or its pharmaceutically acceptable salt.

[0213] The following are exemplary embodiments of compounds represented by formula (I):

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221] And its pharmaceutically acceptable salts.

[0222] In some embodiments, the compounds disclosed herein are pharmaceutically acceptable salts (e.g., HCl salts of compounds of formula I).

[0223] Those skilled in the art will recognize that the kinds listed or exemplified herein are not exhaustive, and additional kinds may be selected within the scope of the terms defined herein.

[0224] Pharmaceutical Composition

[0225] The compositions and methods disclosed herein can be used to treat individuals in need. In some embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal such as a human, the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, a compound disclosed herein and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions, such as water or physiologically buffered saline or other solvents or mediators, such as glycols, glycerols, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are used for human administration, particularly for invasive routes of administration (i.e., routes such as injection or implantation that avoid transport or diffusion across the epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients may be selected, for example, to achieve delayed release of the agent or selective targeting of one or more cells, tissues, or organs. The pharmaceutical compositions may be in the form of dosage units, such as tablets, capsules (including powdered capsules and gelatin capsules), granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system, such as a skin patch. The composition may also be present in a solution suitable for topical application, such as a lotion, cream, or ointment.

[0226] Pharmaceutically acceptable carriers may contain physiologically acceptable agents, such as those used to stabilize, increase solubility, or enhance the absorption of compounds such as those disclosed herein. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier including physiologically acceptable agents depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix in which compounds such as those disclosed herein may be incorporated. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers, and their preparation and administration are relatively simple.

[0227] The phrase “pharmaceutically acceptable” is used in this document to refer to compounds, materials, compositions, and / or dosage forms that, within reasonable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0228] As used herein, the phrase “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and is harmless to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and Soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0229] The pharmaceutical composition (or formulation) may be administered to a subject via any of a variety of routes of administration, including, for example, oral (e.g., oral enemas, tablets, capsules (including powdered capsules and gelatin capsules), granules, powders, pellets, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingual); subcutaneous; transdermal (e.g., as a patch for application to the skin); and topical (e.g., as a cream, ointment, or spray for application to the skin). The compound may also be formulated for inhalation. In some embodiments, the compound may simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and the patents referenced therein.

[0230] The formulation may be conveniently available in a single dosage form and may be prepared by any method well known in the pharmaceutical field. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the specific route of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be the amount of the compound that produces the therapeutic effect. Typically, in 100%, the amount will be in the range of about 1% to about 99% of the active ingredient, preferably about 5% to about 70%, and most preferably about 10% to about 30%.

[0231] Methods for preparing these formulations or compositions include the step of associating an active compound, such as a compound of this disclosure, with a carrier and optionally one or more auxiliary components. Generally, formulations are prepared by homogeneously and tightly associating a compound of this disclosure with a liquid carrier or a fractionated solid carrier, or both, and then, if necessary, shaping the product.

[0232] Formulations of this disclosure suitable for oral administration may be in the form of capsules (including powdered capsules and gelatin capsules), flat capsules, pills, tablets, lozenges (using a flavoring matrix, typically sucrose and gum arabic or tragacanth), lyophilized forms, powders, granules, or as solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil emulsions, or as elixirs or syrups, or as soft lozenges (using an inert matrix, such as gelatin and glycerin, or sucrose and gum arabic) and / or as mouthwashes, each containing a predetermined amount of a compound of this disclosure as an active ingredient. The compositions or compounds may also be administered as large pills, syrups, or pastes.

[0233] To prepare solid dosage forms (capsules (including powdered capsules and gelatin capsules), tablets, pills, sugar-coated pills, powders, granules, etc.) for oral administration, the active ingredient is mixed with one or more pharmaceutically acceptable carriers such as sodium citrate or dicalcium phosphate and / or any of the following: (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders such as, for example, carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and / or gum arabic; (3) humectants such as glycerin; (4) Disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain silicates and sodium carbonate; (5) Solution blockers, such as paraffin; (6) Absorption enhancers, such as quaternary ammonium compounds; (7) Wetting agents, such as, for example, cetyl alcohol and glyceryl monostearate; (8) Absorbents, such as kaolin and bentonite; (9) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) Complexing agents, such as modified cyclodextrin and unmodified cyclodextrin; and (11) Coloring agents. In the case of capsules (including powdered capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain buffers. Similar types of solid compositions may also be used as fillers in soft-filled gelatin capsules and hard-filled gelatin capsules using excipients such as lactose or milk candy and high molecular weight polyethylene glycol.

[0234] Tablets may be made by compression or molding, optionally containing one or more excipients. Compressed tablets may be prepared using binders (e.g., gelatin or hydroxypropyl methylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or croscarmellose sodium), surfactants, or dispersants. Molded tablets may be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0235] Tablets and other solid dosage forms of pharmaceutical compositions, such as sugar-coated pills, capsules (including powdered capsules and gelatin capsules), pellets, and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the field of pharmaceutical formulation. They may also be formulated, for example, using hydroxypropyl methylcellulose in varying proportions to provide a desired release profile, other polymer matrices, liposomes, and / or microspheres to provide slow or controlled release of the active ingredient therein. They may be sterilized by, for example, filtration via a bacterial trap filter, or by incorporating a sterilizing agent in the form of a sterile solid composition, which may be dissolved immediately in sterile water or some other sterile injection medium prior to use. These compositions may also optionally contain light-blocking agents and may be compositions that optionally release the active ingredient in a delayed manner only in or preferably in a portion of the gastrointestinal tract. Examples of encapsulation compositions that may be used include polymeric substances and waxes. If suitable, the active ingredient may also be in a microencapsulated form having one or more of the excipients described above.

[0236] Liquid dosage forms suitable for oral administration include pharmaceutically acceptable emulsions, lyophilized formulations for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrins and their derivatives, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuranol, polyethylene glycol, and fatty acid esters and mixtures thereof of sorbitol.

[0237] In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, coloring agents, aroma agents and preservatives.

[0238] In addition to the active compounds, the suspension may also contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum hydroxide, bentonite, agar and tragacanth gum, and mixtures thereof.

[0239] Dosage forms for topical or transdermal application include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalers. Active compounds may be mixed under sterile conditions with pharmaceutically acceptable carriers and with any preservatives, buffers, or propellants that may be necessary.

[0240] In addition to active compounds, ointments, pastes, creams and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth gum, cellulose derivatives, polyethylene glycol, silicones, bentonite, silicic acid, talc and zinc oxide or mixtures thereof.

[0241] In addition to the active compound, powders and aerosols may also contain excipients such as lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances. Aerosols may also contain commonly used propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0242] Transdermal patches offer the added advantage of controlled delivery of the compounds disclosed herein to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0243] As used herein, the phrases “parenteral administration” and “administered parenterally” refer to administration methods other than enteral and local administration, typically by injection, and including but not limited to intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, spinal, and intrasternal injections and infusions. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds combined with one or more pharmaceutically acceptable sterile isotonic or non-aqueous solutions, dispersions, suspensions, emulsions, or sterile powders, said sterile powders which may be reconstituted into sterile injectable solutions or dispersions prior to use, and which may contain antioxidants, buffers, bacteriostatic agents, solutes or suspending agents or thickeners that make the formulation isotonic with the blood of the intended recipient.

[0244] Examples of suitable aqueous and non-aqueous carriers that may be used in the pharmaceutical compositions of this disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Suitable flowability can be maintained, for example, by using a coating material such as lecithin, by maintaining the desired particle size in the case of a dispersion, and by using a surfactant.

[0245] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers, and dispersants. Antimicrobial activity may be ensured by including various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, and sorbic acid. It may also be desirable to include isotonic agents in the composition, such as sugars and sodium chloride. Furthermore, prolonged absorption of injectable drug forms may be achieved by including agents with delayed absorption, such as aluminum monostearate and gelatin.

[0246] In some cases, to prolong the action of a drug, it is desirable to slow its absorption after subcutaneous or intramuscular injection. This may be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn may depend on the crystal size and crystal form. Alternatively, delayed absorption of parenteral drug forms can be achieved by dissolving or suspending the drug in an oily medium.

[0247] Injectable reservoir formulations are prepared by forming a microencapsulated matrix of the compounds of the invention in a biodegradable polymer such as polylactide-polyglycolic acid. The rate of drug release can be controlled depending on the drug-to-polymer ratio and the properties of the specific polymer used. Examples of other biodegradable polymers include poly(orthoester) and poly(anhydride). Injectable reservoir formulations are also prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

[0248] For use in the methods of this disclosure, the active compound may be administered either on its own or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0249] The introduced methods may also be provided by rechargeable or biodegradable devices. In recent years, various slow-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), both biodegradable and non-biodegradable, can be used to form implants that sustainably release compounds at specific target sites.

[0250] The actual dose level of the active ingredient in a pharmaceutical composition may be altered to obtain an amount of active ingredient that is non-toxic to the patient and effectively achieves the desired therapeutic response for a particular patient, composition, and administration mode.

[0251] The chosen dose level will depend on a variety of factors, including the activity of the specific compound or combination of compounds or its esters, salts or amides used, the route of administration, the time of administration, the excretion rate of the specific compound used, the duration of treatment, other drugs, compounds and / or materials used in combination with the specific compound used, the age, sex, weight, condition, general health and medical history of the patient being treated, and similar factors well known in the medical field.

[0252] A physician or veterinarian with ordinary skill in the art can readily determine and prescribe a therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian may begin with a dose of the pharmaceutical composition or compound at a level below the desired level to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. “Therapeutically effective amount” means the concentration of the compound sufficient to cause the desired therapeutic effect. It should generally be understood that the effective amount of the compound will vary depending on the subject’s weight, sex, age, and medical history. Other factors that may affect the effective amount may include, but are not limited to, the severity of the patient’s condition, the disorder being treated, the stability of the compound, and, if necessary, another type of therapeutic agent administered with the compound disclosed herein. A larger total dose may be delivered by administering the agent multiple times. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al., (1996) Harrison's Principles of Internal Medicine, 13th edition, 1814–1882, which is incorporated herein by reference).

[0253] Generally, a suitable daily dose of the active compound used in the compositions and methods of this disclosure will be the amount of the lowest dose of the compound that effectively produces a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0254] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six, or more sub-dose, which are administered at appropriate intervals throughout the day, optionally in unit dosage form. In some embodiments of this disclosure, the active compound may be administered two or three times daily. In a preferred embodiment, the active compound will be administered once daily.

[0255] Patients receiving this treatment are any animals in need, including primates, especially humans; as well as other mammals such as horses, cattle, pigs, sheep, cats and dogs; poultry; and pets in general.

[0256] In some embodiments, the compounds disclosed herein may be used alone or in combination with another type of therapeutic agent.

[0257] This disclosure includes the use of pharmaceutically acceptable salts of the compounds of this disclosure in the compositions and methods of this disclosure. In some embodiments, the contemplated salts of this disclosure include, but are not limited to, alkylammonium salts, dialkylammonium salts, trialkylammonium salts, or tetraalkylammonium salts. In some embodiments, the contemplated salts of this disclosure include, but are not limited to, L-arginine salts, phenethylbenzylamine salts, benzylamine salts, betaine salts, calcium hydroxide salts, choline salts, dianones salts, diethanolamine salts, diethylamine salts, 2-(diethylamino)ethanol salts, ethanolamine salts, ethylenediamine salts, N-methylglucosamine salts, hebamin salts, 1H-imidazolium salts, lithium salts, L-lysine salts, magnesium salts, 4-(2-hydroxyethyl)morpholine salts, piperazine salts, potassium salts, 1-(2-hydroxyethyl)pyrrolidine salts, sodium salts, triethanolamine salts, tromethamine salts, and zinc salts. In some embodiments, the contemplated salts of this disclosure include, but are not limited to, Na salts, Ca salts, K salts, Mg salts, Zn salts, or other metal salts. In some embodiments, the contemplated salts of this disclosure include, but are not limited to: 1-hydroxy-2-naphthylcarboxylate, 2,2-dichloroacetate, 2-hydroxyethanesulfonate, 2-oxoglutarate, 4-acetamidobenzoate, 4-aminosalicylate, acetate, adipate, L-ascorbate, L-aspartate, benzenesulfonate, benzoate, (+) camphorate, (+) camphor-10-sulfonate, decadecanoate, hexanoate, octanoate, carbonate, cinnamate, citrate, cyclohexanesulfonate, dodecyl sulfate, ethane-1,2-disulfonate, ethanesulfonate, formate, fumarate, mucilage, and other salts. Cholates, d-glucoheponic acid salts, d-glucuronide salts, d-glucuronide salts, glutamate salts, glutamate salts, glycerol phosphate salts, glycolates, hippurate salts, hydrobromide salts, hydrochloride salts, isobutyrate salts, lactates, lactobionate salts, laurate salts, maleate salts, L-malate salts, malonate salts, mandelate salts, methanesulfonate salts, naphthalene-1,5-disulfonate salts, naphthalene-2-sulfonate salts, nicotinate salts, nitrates, oleate salts, oxalate salts, palmitate salts, pamoate salts, phosphates, propionate salts, L-pyroglutamate salts, salicylates, sebacic acid salts, stearates, succinate salts, sulfates, L-tartrate salts, thiocyanates, p-toluenesulfonate salts, trifluoroacetate salts, and undecenoate salts.

[0258] Pharmaceutically acceptable acid addition salts can also exist as various solvates, such as solvates with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates can also be prepared. These solvates can originate from the solvent of crystallization, be inherent in the solvent of preparation or crystallization, or be foreign to such solvents.

[0259] Wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavoring agents and aromas, preservatives, and antioxidants may also be present in the composition. Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0260] In some embodiments, the present invention provides pharmaceutical compositions comprising compounds described herein, such as compounds of formulas I-IV. In some embodiments, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0261] In some embodiments, the pharmaceutical composition may be used to treat or prevent conditions or diseases as described herein.

[0262] The compounds described herein can be used, for example, as therapeutic agents for treating diseases. In some aspects, this disclosure provides pharmaceutical compositions comprising the compounds of this disclosure and pharmaceutically acceptable excipients.

[0263] In some respects, this disclosure provides a method for treating a patient’s disease, the method comprising administering an effective amount of a compound or composition of this disclosure to a patient in need of such treatment.

[0264] In some aspects, this disclosure provides a method of treating an inflammatory disease in a patient, the method comprising administering a compound or composition of the present disclosure to a patient in need. In some aspects, this disclosure provides a method of treating atopic dermatitis or inflammatory bowel disorder in a patient, the method comprising administering a compound or composition of the present disclosure to a patient in need.

[0265] In some aspects, this disclosure provides a method for treating a central nervous system disorder in a patient, the method comprising administering a compound or composition of the present disclosure to a patient in need. In some aspects, this disclosure provides a method for treating a patient with epilepsy, stroke, Alzheimer's disease, schizophrenia, depression, multiple sclerosis, traumatic brain injury, Huntington's disease, or neurodevelopmental disorders, comprising administering a compound or composition of the present disclosure to a patient in need. In some aspects, this disclosure provides a method for treating a patient with a neurodevelopmental disorder (e.g., epilepsy, autism, attention deficit disorder (ADD), or attention deficit hyperactivity disorder (ADHD)), acute neurological disorder (e.g., stroke or traumatic brain injury), neurodegenerative disorder (e.g., Alzheimer's disease, dementia, multiple sclerosis, or Huntington's disease), or psychological / mental disorder (e.g., schizophrenia, depression, anxiety disorder (e.g., generalized anxiety disorder or social anxiety disorder), bipolar disorder, addiction, or obsessive-compulsive disorder (OCD)), the method comprising administering a compound or composition of the present disclosure to a patient in need.

[0266] In some aspects, this disclosure provides methods for treating patients with inflammatory diseases of the lungs, dermatology, and neurology, said methods comprising administering a compound or composition of the present disclosure to a patient in need. In some aspects, this disclosure provides methods for treating patients with asthma, chronic obstructive pulmonary disease (COPD), psoriasis, and rheumatoid arthritis, said methods comprising administering a compound or composition of the present disclosure to a patient in need.

[0267] In some aspects, this disclosure provides methods for improving a patient's cognition, the methods comprising administering a compound or composition of the present disclosure to a patient in need. In some aspects, this disclosure provides methods for treating cognitive and emotional disorders in patients, the methods comprising administering a compound or composition of the present disclosure to a patient in need. In some aspects, this disclosure provides methods for treating learning disabilities, memory loss, or Fragile X syndrome in patients, the methods comprising administering a compound or composition of the present disclosure to a patient in need.

[0268] In some aspects, this disclosure provides a method for increasing the lifespan of a patient. In some aspects, this disclosure provides a method for treating a patient's disease, thereby increasing the patient's lifespan. In some aspects, this disclosure provides a method for increasing the lifespan of a patient, the method comprising administering an effective amount of the compound or composition of this disclosure to a patient in need.

[0269] In some respects, the compounds and pharmaceutical compositions disclosed herein specifically target phosphodiesterases (PDEs), particularly PDE4 isoforms. Therefore, these compounds and pharmaceutical compositions can be used to prevent, reverse, mitigate, or inhibit PDE4 activity. In a preferred embodiment, a method for treating a targeted inflammatory disease is described.

[0270] In some aspects, this disclosure provides a method for treating inflammatory diseases. In some aspects, this disclosure provides methods for treating inflammatory diseases of the lungs, dermatology, and neurology. In some aspects, this disclosure provides methods for treating asthma, chronic obstructive pulmonary disease (COPD), psoriasis, and rheumatoid arthritis. In some aspects, this disclosure provides methods for treating atopic dermatitis or inflammatory bowel disorder.

[0271] In some aspects, this disclosure provides a method for treating central nervous system disorders. In some aspects, this disclosure provides a method for treating epilepsy, stroke, Alzheimer's disease, schizophrenia, depression, multiple sclerosis, traumatic brain injury, Huntington's disease, or neurodevelopmental disorders. In some aspects, this disclosure provides a method for treating neurodevelopmental disorders (e.g., epilepsy, autism, attention deficit disorder (ADD), or attention deficit hyperactivity disorder (ADHD)), acute neurological disorders (e.g., stroke or traumatic brain injury), neurodegenerative disorders (e.g., Alzheimer's disease, dementia, multiple sclerosis, or Huntington's disease), or psychological / mental disorders (e.g., schizophrenia, depression, anxiety disorders (e.g., generalized anxiety disorder or social anxiety disorder), bipolar disorder, addiction, or obsessive-compulsive disorder (OCD)).

[0272] In some aspects, this disclosure provides methods for inhibiting phosphodiesterases (PDEs), particularly PDE4 isoforms. In some aspects, this disclosure provides methods for inhibiting PDE4 and increasing cyclic adenosine monophosphate (cAMP). In some aspects, this disclosure provides methods for inhibiting the hydrolysis of cAMP by inhibiting PDEs such as PDE4.

[0273] In some aspects, this disclosure provides methods for treating cancer. In some embodiments, the cancer is a solid tumor. In some aspects, this disclosure provides methods for treating esophageal cancer, brain cancer, pancreatic cancer, colon cancer, hematologic cancer, lung cancer, prostate cancer, skin cancer, head and neck cancer, CNS cancer, gastric cancer, breast cancer, bladder cancer, or ovarian cancer. In some aspects, this disclosure provides methods for inhibiting tumor growth and metastasis.

[0274] In some aspects, this disclosure provides methods for improving cognition. In some aspects, this disclosure provides methods for treating cognitive and emotional disorders. In some aspects, this disclosure provides methods for treating learning disabilities, memory loss, and Fragile X syndrome.

[0275] In some respects, the compounds disclosed herein are used to treat inflammatory diseases, such as inflammatory diseases of the lungs, inflammatory diseases of the skin, and inflammatory diseases of the neurology. In some respects, the compounds disclosed herein are used to treat asthma, chronic obstructive pulmonary disease (COPD), psoriasis, and rheumatoid arthritis. In some respects, the compounds disclosed herein are used to treat atopic dermatitis or inflammatory bowel disorder.

[0276] In some aspects, the compounds disclosed herein are intended for the treatment of central nervous system disorders. In some aspects, the compounds disclosed herein are intended for the treatment of epilepsy, stroke, Alzheimer's disease, schizophrenia, depression, multiple sclerosis, traumatic brain injury, Huntington's disease, or neurodevelopmental disorders. In some aspects, the compounds disclosed herein are intended for the treatment of neurodevelopmental disorders (e.g., epilepsy, autism, attention deficit disorder (ADD), or attention deficit hyperactivity disorder (ADHD)), acute neurological disorders (e.g., stroke or traumatic brain injury), neurodegenerative disorders (e.g., Alzheimer's disease, dementia, multiple sclerosis, or Huntington's disease), or psychological / mental disorders (e.g., schizophrenia, depression, anxiety disorders (e.g., generalized anxiety disorder or social anxiety disorder), bipolar disorder, addiction, or obsessive-compulsive disorder (OCD)).

[0277] In some aspects, this disclosure provides methods for reducing the frequency of seizures in epilepsy. In some aspects, this disclosure provides methods for treating epilepsy, thereby reducing the frequency of seizures. In some aspects, this disclosure provides methods for reducing the frequency of seizures in epilepsy, wherein said methods further provide a reduction in behavioral side effects and / or cognitive impairments, including but not limited to aggression, irritability, brain fog, memory complications, and language complications.

[0278] In some aspects, the compounds disclosed herein are used to treat inflammatory diseases in patients. In some aspects, the compounds disclosed herein are used to treat inflammatory diseases of the lungs, dermatology, and neurology. In some aspects, the compounds disclosed herein are used to treat asthma, chronic obstructive pulmonary disease (COPD), psoriasis, and rheumatoid arthritis in patients. In some aspects, the compounds disclosed herein are used to treat atopic dermatitis or inflammatory bowel disorder in patients.

[0279] In some aspects, the compounds disclosed herein are used to inhibit PDE4. In some aspects, the compounds disclosed herein are used to inhibit PDE4 and increase cyclic adenosine monophosphate (cAMP). In some aspects, the compounds disclosed herein are used to inhibit the hydrolysis of cAMP by inhibiting PDEs such as PDE4.

[0280] In some respects, the compounds disclosed herein are used to increase lifespan. In some respects, the compounds disclosed herein are used to treat diseases (e.g., epilepsy), thereby increasing lifespan. In some respects, the compounds disclosed herein are used to treat diseases (e.g., epilepsy), thereby improving cognition.

[0281] In some respects, the compounds disclosed herein are used to treat cancer. In some respects, the compounds disclosed herein are used to treat esophageal cancer, brain cancer, pancreatic cancer, and colon cancer. In some respects, the compounds disclosed herein are used to inhibit tumor growth and metastasis.

[0282] In some aspects, the compounds disclosed herein are used to improve cognition. In some aspects, the compounds disclosed herein are used to treat cognitive and emotional disorders. In some aspects, the compounds disclosed herein are used to treat learning disabilities, memory loss, and Fragile X syndrome. In some aspects, the compounds disclosed herein are used to improve cognition in non-disease models.

[0283] In some aspects, this disclosure provides a method for treating mammals suffering from inflammatory diseases. In some aspects, this disclosure provides a method for treating mammals suffering from inflammatory diseases of the lungs, dermatology, and neurology. In some aspects, this disclosure provides a method for treating mammals suffering from asthma, chronic obstructive pulmonary disease (COPD), psoriasis, and rheumatoid arthritis. In some aspects, this disclosure provides a method for treating mammals suffering from atopic dermatitis or inflammatory bowel disorder.

[0284] In some aspects, this disclosure provides a method for treating mammals suffering from central nervous system disorders. In some aspects, this disclosure provides a method for treating mammals suffering from epilepsy, stroke, Alzheimer's disease, schizophrenia, depression, multiple sclerosis, traumatic brain injury, Huntington's disease, or neurodevelopmental disorders. In some aspects, this disclosure provides a method for treating mammals suffering from epilepsy, thereby reducing the frequency of seizures. In some aspects, this disclosure provides a method for treating mammals suffering from neurodevelopmental disorders (e.g., epilepsy, autism, attention deficit disorder (ADD), or attention deficit hyperactivity disorder (ADHD)), acute neurological disorders (e.g., stroke or traumatic brain injury), neurodegenerative disorders (e.g., Alzheimer's disease, dementia, multiple sclerosis, or Huntington's disease), or psychological / mental disorders (e.g., schizophrenia, depression, anxiety disorders (e.g., generalized anxiety disorder or social anxiety disorder), bipolar disorder, addiction, or obsessive-compulsive disorder (OCD)).

[0285] In some aspects, this disclosure provides a method for treating mammals suffering from cognitive or emotional disorders. In some aspects, this disclosure provides a method for treating mammals suffering from learning disabilities, memory loss, and Fragile X syndrome. In some aspects, this disclosure provides a method for improving cognition in non-disease models.

[0286] In some aspects, this disclosure provides methods for treating mammals suffering from cancer. In other aspects, this disclosure provides methods for treating mammals suffering from esophageal cancer, brain cancer, pancreatic cancer, and colon cancer.

[0287] In some embodiments, the compounds disclosed herein are prodrugs of the compounds described herein. For example, the hydroxyl group in the parent compound is present in the form of an ester or carbonate, or the carboxylic acid in the parent compound is present in the form of an ester. In some such embodiments, the prodrug is metabolized in vivo to the active parent compound (e.g., the ester is hydrolyzed to the corresponding hydroxyl or carboxylic acid).

[0288] In some embodiments, the compounds of this disclosure may be racemic. In some embodiments, the compounds of this disclosure may be rich in one enantiomer. For example, the compounds of this disclosure may have greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, or even 95% or higher of ee. In some embodiments, the compounds of this disclosure may have more than one stereocenter. In some such embodiments, the compounds of this disclosure may be rich in one or more diastereomers. For example, the compounds of this disclosure may have greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, or even 95% or higher of de.

[0289] Chemical synthesis methods

[0290] The following examples are provided to illustrate, but not to limit, this disclosure. Those skilled in the art will recognize that the following synthetic reactions and schemes can be modified by selecting suitable starting materials and reagents to obtain other compounds of formulas I, II, III, and IV.

[0291] Abbreviations: The embodiments described herein use materials including, but not limited to, those described by the following abbreviations known to those skilled in the art:

[0292]

[0293]

[0294] The proposed target compound can be prepared by conventional chemical methods or according to the general scheme shown below.

[0295] Synthesis of benzothiazole PDE4 inhibitors

[0296] Reagents were purchased at least reagent grade from Aldrich, Acros, Alfa Aesar, or other suppliers and used without further purification. Compounds were synthesized according to the general methods described below, and their purity was determined by HPLC.

[0297] The compounds were synthesized using the Hugerschoff reaction to synthesize benzothiazole heterocycles, with amide coupling introducing the R2 substituent and Suzuki cross-coupling introducing the R1 substituent.

[0298] Scheme 1. Representative synthesis of the benzothiazole compounds disclosed herein.

[0299]

[0300]

[0301] a. KSCN (4 equivalents) / Br2 (1 equivalent), AcOH, room temperature, 20 h (96%); b. NBS (1.5 equivalents), THF, room temperature, 20 h (90%); c. DMAP (1.2 equivalents), di-tert-butyl dicarbonate (1.5 equivalents), DCM, room temperature, 20 h; d. LiOH (4 equivalents), MeOH / water, 60℃, 20 h (95%); e. 3-aminopyridine (1.2 equivalents), EDC.HCl (1.2 equivalents), DMAP (0.8 equivalents), THF, 20 h (67%); f. 3-substituted boric acid (1.2 equivalents), cesium carbonate (1.5 equivalents), bis(triphenylphosphine)palladium(II) dichloride (0.15 equivalents), dioxane, EtOH-H2O (1:1) (50%-70%); g. 4M HCl / dioxane; h. tert-butyl nitrite, tetrahydrofuran (THF), 60°C for 16 h, (15-32%); i. 4M HCl / dioxane, ethyl acetate, (90%).

[0302] Table 1. Benzothiazole analogues disclosed herein.

[0303]

[0304]

[0305] Table 2. Synthesized benzothiazole analogs of this disclosure.

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313] intermediate synthesis

[0314] Preparation of (4-bromo-6-(pyridin-3-ylcarbamoyl)benzothiazo-2-yl)tert-butyl carbamate (I-5)

[0315]

[0316] Step 1: Ethyl 2-aminobenzothiazole-6-carboxylate

[0317]

[0318] Ethyl 4-aminobenzoate (50 mmol, 8.25 g) was dissolved in acetic acid (200 ml). Potassium thiocyanate (KSCN) (200 mmol, 19.43 g) was added to the mixture. Bromine (Br2) (50 mmol, 2.56 ml) was slowly added to the mixture over 30 minutes. The reaction mixture was then stirred at room temperature for 20 h. The reaction mixture was then poured into ice-cold water and alkalized to pH 8 using ammonium hydroxide (NH4OH). The precipitate was then extracted with ethyl acetate and water. The organic layer was separated, dried, and evaporated to give ethyl 2-aminobenzothiazole-6-carboxylate (10.78 g, 97% yield) as a pale yellow solid. TLC (SiO2, EtOAc / hexane 1:1) R f = 0.38; 1 H NMR (500 MHz, DMSO) δ 8.29 (d, J = 1.8 Hz, 1H), 7.92 (s, 2H), 7.82 (dd, J = 8.5, 1.8 Hz, 1H), 7.38 (d, J = 8.5 Hz, 1H), 4.29 (q, J = 7.1 Hz, 2H), 1.31 (t, J = 7.1 Hz, 3H). 13C NMR (125 MHz, DMSO) δ170.21, 166.11, 157.32, 131.60, 127.52, 122.96, 122.45, 117.53, 60.82, 14.72.

[0319] Step 2: Ethyl 2-amino-4-bromobenzothiazole-6-carboxylate (I-2)

[0320]

[0321] Ethyl 2-aminobenzothiazole-6-carboxylate (20 mmol, 4.45 g) was dissolved in acetic acid (100 mL). Bromine (Br2) (40 mmol, 2.05 mL) in 20 mL of acetic acid was slowly added to the mixture over 30 minutes. The reaction mixture was then stirred at 80 °C for 20 h. The reaction mixture was then poured into ice-cold water and alkalized to pH 8 using ammonium hydroxide (NH4OH). The precipitate was then extracted with ethyl acetate and water. The organic layer was separated, dried, and evaporated to give ethyl 2-amino-4-bromobenzothiazole-6-carboxylate (5.06 g, 84% yield) as a pale orange solid. TLC (SiO2, EtOAc / hexane 1:1) R f = 0.58; 1 H NMR (500 MHz, DMSO) δ 8.30 (d, J = 1.6 Hz, 1H), 7.95 (d, J =1.6 Hz, 1H), 7.51 (s, 2H), 4.29 (q, J = 7.1 Hz, 2H), 1.31 (t, J = 7.1 Hz, 3H). 13 C NMR (125 MHz, DMSO) δ 170.73, 164.97, 154.01, 131.54, 130.09, 123.76, 122.51, 109.64, 61.29, 14.65.

[0322] Step 3: Ethyl 4-bromo-2-((tert-butoxycarbonyl)amino)benzothiazole-6-carboxylate

[0323]

[0324] Ethyl 2-amino-4-bromobenzothiazol-6-carboxylate (5 mmol, 1.5 g) was dissolved in dichloromethane (100 ml). 4-(diethylamino)pyridine (6 mmol, 733 mg) was added to the reaction mixture. Di-tert-butyl dicarbonate (10 mmol, 2.18 g) was added to the mixture. The reaction mixture was stirred at room temperature for 20 h. After 20 h, the solvent was evaporated, and the crude product was purified using 20% ​​ethyl acetate / dichloromethane to give ethyl 4-bromo-2-((tert-butoxycarbonyl)amino)benzothiazol-6-carboxylate (1.8 g, 90% yield) as a white solid. TLC (SiO2, EtOAc / hexane 1:1) R f = 0.68; 1 H NMR(500 MHz, DMSO) δ 12.40 (s, 1H), 8.62 (d, J = 1.6 Hz, 1H), 8.12 (d, J = 1.6Hz, 1H), 4.35 (q, J = 7.1 Hz, 2H), 1.52 (s, 9H), 1.35 (t, J = 7.1 Hz, 3H). 13 CNMR (126 MHz, DMSO) δ 164.93, 164.54, 153.40, 151.41, 133.03, 130.05, 125.95, 123.45, 113.13, 82.88, 28.25, 14.65.

[0325] Step 4: 4-Bromo-2-((tert-butoxycarbonyl)amino)benzothiazol-6-carboxylic acid

[0326]

[0327] Ethyl 4-bromo-2-((tert-butoxycarbonyl)amino)benzothiazol-6-carboxylic acid (4 mmol, 1.60 g, 1 equivalent) was dissolved in 50 mL of methanol. Lithium hydroxide (LiOH) (16 mmol, 383 mg, 4 equivalent) was dissolved in 10 mL of deionized water. The lithium hydroxide (LiOH) solution was added to the reaction mixture. The reaction mixture was then stirred at 60 °C for 16 h. The reaction mixture was then cooled, and the solvent was evaporated. The residue was then dissolved in 100 mL of deionized water and acidified to pH 4 with 1 M HCl. The precipitate formed was filtered off by vacuum filtration to give 4-bromo-2-((tert-butoxycarbonyl)amino)benzothiazol-6-carboxylic acid (1.35 g, 91% yield) as a white solid. TLC (SiO2, EtOAc / hexane 1:1) R f =0.28;1 H NMR (500 MHz, DMSO) δ 12.36 (s, 1H), 8.57 (d, J = 1.6 Hz, 1H), 8.09 (d, J = 1.5 Hz, 1H), 1.50 (s, 9H). 13 C NMR (126 MHz, DMSO) δ 166.42, 164.19, 151.12, 132.88, 130.32, 126.99, 123.50, 113.01, 82.79, 28.24.

[0328] Step 5: (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazo-2-yl)tert-butyl carbamate (I-5)

[0329]

[0330] 4-Bromo-2-((tert-butoxycarbonyl)amino)benzothiazol-6-carboxylic acid (8.45 mmol, 3 g) was dissolved in tetrahydrofuran (THF) (100 ml). 4-(dimethylamino)pyridine (6.76 mmol, 825 mg) and EDC.HCl (12.87 mmol, 2.43 g) were added to the mixture. 3-Aminopyridine (10 mmol, 941 mg) was added to the mixture. The reaction mixture was stirred at room temperature under an inert atmosphere for 20 h. The solvent was then evaporated, and the crude product was extracted with ethyl acetate / water. The ethyl acetate layer was dried, evaporated, and the crude product was purified by silica using 10% methanol in dichloromethane to give tert-butyl (4-bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)carbamate (3.07 g, 81% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1∶9) R f = 0.58; 1 H NMR (500 MHz, DMSO) δ 12.39 (s, 1H),10.55 (s, 1H), 8.95 (d, J = 2.6 Hz, 1H), 8.57 (d, J = 1.7 Hz, 1H), 8.33 (dd,J = 4.7, 1.5 Hz, 1H), 8.25 – 8.19 (m, 2H), 7.42 (dd, J = 8.3, 4.7 Hz, 1H), 1.51 (s, 9H). 13C NMR (126 MHz, DMSO) δ 169.88, 164.71, 150.75, 145.08, 142.47,142.39, 136.27, 132.89, 128.89, 127.80, 124.03, 123.99, 121.58, 112.78,82.10, 28.34.

[0331] Preparation of 4-bromo-6-(bromomethyl)benzothiazole (I-8)

[0332]

[0333] Step 1: Ethyl 4-bromobenzothiazole-6-carboxylate (I-6)

[0334]

[0335] Ethyl 2-amino-4-bromobenzothiazol-6-carboxylate (15 mmol, 4.51 g) was dissolved in dioxane (50 ml). Tert-butyl nitrite (22.5 mmol, 2.65 ml) was added to the reaction mixture. The reaction mixture was stirred at 60 °C for 4 h. The solvent was then evaporated, and the crude product was extracted with ethyl acetate and water. The organic layer was separated, dried, evaporated, and purified by silica using 40% ethyl acetate / hexane to give ethyl 4-bromobenzothiazol-6-carboxylate (3.02 g, 47% yield) as an orange solid. TLC (SiO2, EtOAc / hexane 1:4) R f = 0.48; 1 H NMR (500 MHz, CDCl3) δ 9.25 (s,1H), 8.66 (d, J = 1.5 Hz, 1H), 8.42 (d, J = 1.5 Hz, 1H), 4.46 (q, J = 7.1 Hz,2H), 1.46 (t, J = 7.2 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 164.90, 157.76, 154.08, 134.49, 130.60, 129.01, 123.27, 117.19, 61.81, 14.34.

[0336] Step 2: (4-bromobenzothiazol-6-yl)methanol

[0337]

[0338] Ethyl 4-bromobenzothiazol-6-carboxylate (3 mmol, 858 mg) was dissolved in 30 mL of tetrahydrofuran (THF). Diisobutylaluminum hydrogenate (1 M, in hexane) (12 mL) was added at -70 °C under an inert atmosphere. The reaction mixture was then stirred at room temperature for 16 h. Saturated potassium sodium tartrate (4 mL) and water (4 mL) were added to the reaction mixture. The reaction mixture was stirred for 1 h and extracted with ethyl acetate and water. The organic phase was separated, dried, evaporated, and purified by silica using 10% ethyl acetate / dichloromethane to give (4-bromobenzothiazol-6-yl)methanol (0.25 g, 34% yield) as a pale orange solid. TLC (SiO2, EtOAc / DCM 1:9) R f = 0.26; 1 H NMR (300 MHz, DMSO) δ 7.58 (d, J =1.7 Hz, 1H), 7.41 (s, 1H), 7.28 (d, J = 8.1 Hz, 1H), 7.15 (dd, J = 8.2, 1.7Hz, 1H), 4.49 (s, 2H). 13 C NMR (75 MHz, DMSO) δ 166.65, 152.21, 135.77, 131.21, 124.87, 119.54, 117.71, 63.48.

[0339] Step 3: 4-Bromo-6-(bromomethyl)benzothiazole (I-8)

[0340]

[0341] (4-Bromobenzothiazol-6-yl)methanol (1 mmol, 244 mg) was dissolved in 25 mL diethyl ether. Phosphorus tribromide (PBr3) (1.5 mmol, 0.15 mL) in 10 mL diethyl ether was slowly added to the reaction mixture. The reaction mixture was stirred overnight at room temperature. The solvent was then evaporated, and the residue was extracted with ethyl acetate and water. The ethyl acetate layer was separated, dried, and evaporated to give 4-bromo-6-(bromomethyl)benzothiazolazole (200 mg, 65% yield) as a pale yellow solid. TLC (SiO2, EtOAc / DCM 2:3) R f = 0.53; 1 H NMR (500 MHz, CDCl3) δ 9.12 (s, 1H), 7.96,7.96, (s, 1H), 7.80, 7.80 (s, 1H), 4.61 (s, 2H). 13C NMR (126 MHz, CDCl3) δ155.49, 151.42, 136.77, 134.91, 130.79, 121.51, 117.45, 31.8.

[0342] Preparation of ethyl 4-(3-chlorophenyl)benzothiazole-6-carboxylate (I-9)

[0343]

[0344] Ethyl 4-bromobenzothiazol-6-carboxylate (13 mmol, 3.71 g) was dissolved in toluene (30 ml) ethanol / water (10 ml 1:1), and 3-chlorophenylboronic acid (15.6 mmol, 2.43 g) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.78 mmol, 901 mg) and potassium carbonate K2CO3 (26 mmol, 3.59 g) were added to the reaction mixture. The reaction mixture was stirred at 80 °C for 20 h under argon. The solvent was then evaporated and extracted with ethyl acetate. The crude product was purified by silica using 30% ethyl acetate / hexane to give ethyl 4-(3-chlorophenyl)benzothiazol-6-carboxylate (2.89 g, 70% yield) as a white solid. TLC (SiO2, EtOAc / hexane 2:3) R f = 0.48; 1 H NMR (500 MHz, CDCl3) δ 9.21 (s, 1H), 8.73 (d, J = 1.6 Hz, 1H), 8.28 (d, J = 1.6 Hz, 1H), 7.88 (t, J = 1.9 Hz, 1H), 7.77 (dt, J = 7.5, 1.5 Hz, 1H), 7.51 – 7.41 (m,2H), 4.49 (q, J = 7.1 Hz, 2H), 1.48 (t, J = 7.1 Hz, 3H). 13 C NMR (126 MHz, CDCl3) δ 165.93, 156.99, 153.43, 139.79, 135.18, 135.01, 134.33, 129.71,129.64, 128.19, 128.04, 127.85, 127.33, 123.53, 61.56, 14.41.

[0345] Compound Synthesis

[0346] Preparation of 2-amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (compound 1)

[0347]

[0348] Step 1: 2-Amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide

[0349] (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)carbamate tert-butyl ester (0.9 mmol, 404 mg) was dissolved in dioxane (8 ml) ethanol / water (4 ml 1:1), and 3-chlorophenylboronic acid (1 mmol, 156 mg) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.1 mmol, 71 mg) and cesium carbonate Cs2CO3 (1 mmol, 325 mg) were added to the reaction mixture. The reaction mixture was stirred at 160 °C under argon in a Biotage microwave initiator for 30 min. The solvent was then evaporated and extracted with ethyl acetate. The crude product was purified by silica using 50% ethyl acetate / hexane to give 2-amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (140 mg, 41% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1:9) R f = 0.38; HPLC - 96.42%. 1 HNMR (500 MHz, DMSO) δ 10.38 (s, 1H), 8.94 (dd, J = 2.6, 0.8 Hz, 1H), 8.34 (d,J = 1.8 Hz, 1H), 8.31 (dd, J = 4.7, 1.5 Hz, 1H), 8.20 (ddd, J = 8.3, 2.6, 1.5Hz, 1H), 8.01 (s, 1H), 8.01 (s, 2H), 7.93 (t, J = 1.9 Hz, 1H), 7.81 (dt, J =7.9, 1.2 Hz, 1H), 7.52 (t, J = 7.9 Hz, 1H), 7.48 – 7.37 (m, 2H). 13C NMR (126MHz, DMSO) δ 169.61, 165.78, 153.50, 144.87, 142.48, 141.51, 136.47, 133.19,132.74, 130.40, 129.37, 128.37, 128.15, 127.78, 127.52, 127.17, 126.07,123.99, 121.22. HRMS [M+H] + = 381.0571; [M+Na] + = 403.0391.

[0350] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (Compound 1)

[0351] To a solution of (4-(3-chlorophenyl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (115 mg, 0.24 mmol) in 1,4-dioxane (20 mL), 4 M HCl in 1,4-dioxane was added, and the reaction was stirred overnight at 60 °C. The mixture was concentrated under reduced pressure to give the product as a white solid (96 mg, 96% yield). 1 H NMR (300 MHz, DMSO-d6) δ 9.58 (s, 1H), 8.81 (m, 1H), 8.59 (m,1H), 8.44 (m, 1H), 8.06–8.04 (m, 1H), 7.89–7.76 (m, 2H), 7.43–7.32 (m, 2H).

[0352] Preparation of 2-amino-4-(3-cyanophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide (compound 2)

[0353]

[0354] (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)carbamate tert-butyl ester (0.67 mmol, 300 mg) was dissolved in dioxane (8 ml) ethanol / water (4 ml 1:1), and 3-cyanophenylboronic acid (0.8 mmol, 146 mg) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.1 mmol, 71 mg) and cesium carbonate Cs2CO3 (1 mmol, 325 mg) were added to the reaction mixture. The reaction mixture was stirred at 160 °C under argon in a Biotage microwave initiator for 30 min. The solvent was then evaporated and extracted with ethyl acetate. The crude product was washed with methanol to give 2-amino-4-(3-cyanophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (180 mg, 72% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1:9) R f = 0.32; HPLC - 97%; 1 H NMR (500 MHz, DMSO) δ 10.40 (s, 1H), 8.94 (d, J = 2.6 Hz, 1H), 8.38 – 8.33 (m, 2H), 8.32 (dd, J = 4.7, 1.5 Hz, 1H), 8.24 – 8.18 (m, 2H), 8.06 (d, J = 2.0 Hz, 3H), 7.87 – 7.84 (m, 1H), 7.71 (t, J = 7.8 Hz, 1H), 7.43 – 7.39 (m, 1H). 13 C NMR(126 MHz, DMSO) δ 169.85, 165.74, 153.50, 144.90, 142.48, 140.50, 136.45,134.49, 133.13, 132.79, 131.30, 129.87, 127.79, 127.34, 127.26, 126.11,124.01, 121.55, 119.51, 111.66. HRMS [M+H] + = 372.0914; [M+Na] + = 394.0733.

[0355] Preparation of 2-amino-4-(3-(methanesulfonyl)phenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide (compound 3) and 2-amino-4-(3-(methanesulfonyl)phenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (compound 7)

[0356]

[0357] Step 1: 2-Amino-4-(3-(methanesulfonyl)phenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide (Compound 3)

[0358] (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)carbamate tert-butyl ester (0.67 mmol, 300 mg) was dissolved in dioxane (8 ml) ethanol / water (4 ml 1:1), and 3-methylsulfonylphenylboronic acid (0.8 mmol, 160 mg) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.1 mmol, 71 mg) and cesium carbonate Cs2CO3 (1 mmol, 325 mg) were added to the reaction mixture. The reaction mixture was stirred at 160 °C under argon in a Biotage microwave initiator for 30 min. The solvent was then evaporated and extracted with ethyl acetate. The crude product was washed with methanol to give 2-amino-4-(3-methylsulfonyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (200 mg, 71% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1:9) R f = 0.42; HPLC - 96.86%; 1 H NMR (500MHz, DMSO) δ 10.42 (s, 1H), 8.95 (d, J = 2.6 Hz, 1H), 8.38 (d, J = 1.8 Hz, 1H), 8.34 (t, J = 1.8 Hz, 1H), 8.32 (dd, J = 4.7, 1.5 Hz, 1H), 8.21 (ddd, J =8.3, 2.8, 1.5 Hz, 2H), 8.04 (d, J = 1.9 Hz, 1H), 8.00 (s, 2H), 7.96 – 7.92(m, 1H), 7.77 (t, J = 7.8 Hz, 1H), 7.41 (dd, J = 8.4, 4.7 Hz, 1H), 3.29 (s, 3H). 13C NMR (126 MHz, DMSO) δ 169.72, 165.84, 153.58, 144.88, 142.43, 141.26,140.56, 136.48, 134.85, 132.70, 129.65, 128.01, 127.74, 127.31, 126.28,126.02, 124.01, 121.40, 55.38, 44.11. HRMS [M+H] + = 425.0737; [M+Na] + =447.0556.

[0359] Step 2: 2-Amino-4-(3-(methanesulfonyl)phenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 7)

[0360] Compound 7 was prepared using a similar method as described in step 2 of the preparation of compound 1.

[0361] Preparation of 2-amino-4-(5-chlorothiophen-2-yl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide (compound 4) and 2-amino-4-(5-chlorothiophen-2-yl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (compound 8)

[0362]

[0363] Step 1: 2-Amino-4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (Compound 4)

[0364] (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)tert-butyl carbamate (0.45 mmol, 200 mg) was dissolved in dioxane (8 ml) ethanol / water (4 ml 1:1), and 5-chlorothiophene-2-boronic acid (0.53 mmol, 87 mg) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.08 mmol, 47 mg) and cesium carbonate Cs2CO3 (0.68 mmol, 217 mg) were added to the reaction mixture. The reaction mixture was stirred at 160 °C under argon atmosphere in a Biotage microwave initiator for 30 min. The solvent was then evaporated and extracted with ethyl acetate. The crude product was purified over silica using 50% ethyl acetate / hexane to give 2-amino-4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (103 mg, 60% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1:9) R f =0.46; HPLC- 95.69%. 1 H NMR (500 MHz, DMSO) δ 10.42 (s, 1H), 8.95 (d, J = 2.6Hz, 1H), 8.32 (dd, J = 4.7, 1.5 Hz, 1H), 8.29 (d, J = 1.8 Hz, 1H), 8.25 (d, J= 1.7 Hz, 1H), 8.21 (ddd, J = 8.4, 2.6, 1.5 Hz, 1H), 8.16 (s, 2H), 7.80 (d, J= 4.1 Hz, 1H), 7.42 (dd, J = 8.3, 4.7 Hz, 1H), 7.21 (d, J = 4.1 Hz, 1H) 13 CNMR (126 MHz, DMSO) δ 169.58, 165.76, 151.20, 144.92, 142.49, 139.14, 136.44,132.42, 129.66, 127.78, 127.31, 126.87, 124.85, 124.01, 122.23, 121.64,120.35. HRMS [M+H] + = 387.0136; [M+Na] + = 408.9955.

[0365] Step 2: 2-Amino-4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 8)

[0366] Compound 8 was prepared using a similar method as described in step 2 of the preparation of compound 1.

[0367] Preparation of 2-amino-4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide (compound 5) and 2-amino-4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (compound 10)

[0368]

[0369] Step 1: 2-Amino-4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide (Compound 5)

[0370] (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)carbamate tert-butyl ester (0.9 mmol, 404 mg) was dissolved in dioxane (8 ml) ethanol / water (4 ml 1:1), and 3-nitrophenylboronic acid (1 mmol, 156 mg) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.1 mmol, 71 mg) and cesium carbonate Cs2CO3 (1 mmol, 325 mg) were added to the reaction mixture. The reaction mixture was stirred at 160 °C under argon in a Biotage microwave initiator for 30 min. The solvent was then evaporated and extracted with ethyl acetate. The crude product was purified by silica using hexane / ethyl acetate to give 2-amino-4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (100 mg, 57% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1:9) R f = 0.42; HPLC - 95.25%; 1H NMR (500 MHz, DMSO) δ 10.43 (s, 1H), 8.94 (d, J = 2.6 Hz, 1H), 8.73 (t, J= 2.0 Hz, 1H), 8.39 (d, J = 1.8 Hz, 1H), 8.32 (td, J = 5.7, 1.5 Hz, 2H), 8.23(dddd, J = 22.3, 8.4, 2.5, 1.3 Hz, 2H), 8.08 (d, J = 1.8 Hz, 1H), 8.06 (s,2H), 7.80 (t, J = 8.0 Hz, 1H), 7.41 (dd, J = 8.3, 4.7 Hz, 1H). 13 C NMR (126MHz, DMSO) δ 169.95, 165.75, 153.51, 148.19, 144.91, 142.46, 140.92, 136.44,136.27, 132.82, 130.15, 127.78, 127.30, 127.12, 126.19, 124.17, 124.02,122.48, 121.68. HRMS [M+H] + = 392.0812; [M+Na] + = 414.0631.

[0371] Step 2: 2-Amino-4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 10)

[0372] Compound 10 is prepared according to a similar method described in step 2 of the preparation of compound 1.

[0373] Preparation of 4-(3-acetylphenyl)-2-amino-N-(pyridin-3-yl)benzothiazole-6-carboxamide (compound 6) and 4-(3-acetylphenyl)-2-amino-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (compound 9)

[0374]

[0375] Step 1: 4-(3-acetylphenyl)-2-amino-N-(pyridin-3-yl)benzothiazole-6-carboxamide (compound 6)

[0376] (4-Bromo-6-(pyridin-3-ylcarbamoyl)benzothiazol-2-yl)carbamate tert-butyl ester (0.45 mmol, 200 mg) was dissolved in dioxane (8 ml) ethanol / water (4 ml 1:1), and 3-acetylphenylboronic acid (0.53 mmol, 88 mg) was added to the mixture. Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh3)2Cl2) (0.08 mmol, 47 mg) and cesium carbonate Cs2CO3 (0.68 mmol, 217 mg) were added to the reaction mixture. The reaction mixture was stirred at 160 °C under argon in a Biotage microwave initiator for 30 min. The solvent was then evaporated and extracted with ethyl acetate. The crude product was washed with methanol to give 2-amino-4-(3-acetylphenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (56 mg, 33% yield) as a pale yellow solid. TLC (SiO2, MeOH / DCM 1:9) R f = 0.42; HPLC - 95%; 1 H NMR (500MHz, DMSO) δ 10.41 (s, 1H), 8.95 (d, J = 2.6 Hz, 1H), 8.37 (t, J = 1.8 Hz,1H), 8.36 (d, J = 1.9 Hz, 1H), 8.31 (dd, J = 4.7, 1.5 Hz, 1H), 8.21 (ddd, J =8.4, 2.6, 1.5 Hz, 1H), 8.12 – 8.09 (m, 1H), 8.02 (d, J = 1.9 Hz, 1H), 7.98(s, 3H), 7.64 (t, J = 7.7 Hz, 1H), 7.43 – 7.39 (m, 1H), 2.67 (s, 3H). 13 C NMR(126 MHz, DMSO) δ 198.56, 169.49, 165.88, 153.65, 144.85, 142.44, 139.88,137.23, 136.50, 134.49, 132.63, 129.33, 129.02, 128.94, 127.72, 127.48,127.24, 126.17, 124.00, 121.02, 27.43. HRMS [M+H] + = 389.1067; [M+Na] + =411.0886.

[0377] Step 2: 4-(3-acetylphenyl)-2-amino-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 9)

[0378] Compound 9 was prepared using a similar method as described in step 2 of the preparation of compound 1.

[0379] Preparation of 4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (compound 11)

[0380]

[0381] Step 1: 4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide

[0382] 2-Amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (0.263 mmol, 100 mg) was dissolved in THF (50 mL). Tert-butyl nitrite (0.53 mmol, 0.063 mL) was added to the reaction mixture, and the mixture was stirred at 60 °C for 16 h. The solvent was then evaporated, the crude product was extracted with ethyl acetate, and the ethyl acetate layer was dried and evaporated. The residue was dissolved in dichloromethane (5 mL), and hexane (5 mL) was added to precipitate 4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (31 mg, 32% yield) as a pale brown solid. 1 H NMR (500MHz, acetone) δ 10.07 (s, 1H), 9.53 (s, 1H), 9.02 (s, 1H), 8.86 (d, J = 1.4 Hz,1H), 8.39 (d, J = 1.9 Hz, 1H), 8.38 (s, 1H), 8.34 (d, J = 2.6 Hz, 1H), 8.34(d, J = 0.8 Hz, 1H), 8.07 (t, J = 1.8 Hz, 1H), 7.96 (d, J = 2.7 Hz, 1H), 7.94(d, J = 2.7 Hz, 1H), 7.61 (t, J = 15.5 Hz, 1H), 7.53 (d, J = 3.2 Hz, 1H),7.43 (d, J = 13.0 Hz, 1H). 13C10 NMR (126 MHz, acetone) δ 165.13, 158.05, 152.73, 144.73, 141.84, 141.76, 140.19, 135.63, 134.39, 133.58, 132.23, 129.91, 129.67, 128.26, 127.87, 127.21, 127.12, 125.30, 123.48, 122.16. HRMS [M+H] + = 366.0462; [M+Na] + = 388.0282.

[0383] Step 2: 4-(3-chlorophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 11)

[0384] 4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (30 mg, 0.08 mmol) was dissolved in EtOAc (10 mL) and 4M HCl was added to 1,4-dioxane (1 mL). The solution was stirred at room temperature for 2 h. The solvent was then removed under reduced pressure to give the product 4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide hydrochloride (25 mg, 78% yield) as a solid.

[0385] Preparation of 4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide HCl salt (compound 12)

[0386]

[0387] Step 1: 4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide

[0388] 2-Amino-4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (0.256 mmol, 100 mg) was dissolved in THF (50 mL). Tert-butyl nitrite (0.53 mmol, 0.063 mL) was added to the reaction mixture, and the mixture was stirred at 60 °C for 16 h. The solvent was then evaporated, the crude product was extracted with ethyl acetate, and the ethyl acetate layer was dried and evaporated. The residue was dissolved in dichloromethane (5 mL), and hexane (5 mL) was added to precipitate 4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (30 mg, 33% yield) as a pale brown solid. 1H NMR(500 MHz, acetone) δ 10.12 (s, 1H), 9.59 (s, 1H), 9.05 (d, J = 2.2 Hz, 1H), 8.93(d, J = 1.7 Hz, 2H), 8.47 (d, J = 2.7 Hz, 1H), 8.46 (d, J = 1.7 Hz, 1H), 8.42(d, J = 3.6 Hz, 1H), 8.37 (dd, J = 1.3, 8.2 Hz, 1H), 7.89 (t, J = 8.0 Hz,1H), 7.49 (dd, J = 8.2 Hz, 1H). 13 C10 NMR (126 MHz, acetone) δ 165.04, 158.57, 152.71, 148.40, 144.55, 141.53, 139.72, 135.91, 135.76, 133.42, 132.35, 129.65, 125.50, 124.47, 123.61, 122.68. HRMS [M+H] + = 377.0703; [M+Na] + =399.0522.

[0389] Step 2: 4-(3-nitrophenyl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 12)

[0390] Compound 12 is prepared according to a similar method described in step 2 of the preparation of compound 11.

[0391] Preparation of 4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (compound 13)

[0392]

[0393] Step 1: 4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazole-6-carboxamide

[0394] 2-Amino-4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (0.26 mmol, 100 mg) was dissolved in THF (50 ml). Tert-butyl nitrite (0.53 mmol, 0.063 ml) was added to the reaction mixture, and the mixture was stirred at 60 °C for 16 h. The solvent was then evaporated, the crude product was extracted with ethyl acetate, and the ethyl acetate layer was dried and evaporated. The residue was dissolved in dichloromethane (5 ml), and hexane (5 ml) was added to precipitate 4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide (15 mg, 15% yield) as a pale brown solid. 1 H NMR (500 MHz, acetone) δ 10.02 (s, 1H), 9.59 (s, 1H), 9.01 (d, J = 2.0 Hz,1H), 8.74 (d, J = 1.5 Hz, 1H), 8.55 (d, J = 1.6 Hz, 1H), 8.38 (d, J = 1.3 Hz,1H), 8.36 (d, J = 2.5 Hz, 1H), 7.96 (d, J = 4.1 Hz, 1H), 7.42 (dd, J = 4.7,8.2 Hz, 1H), 7.19 (d, J = 4.0 Hz, 1H). 13 C10 NMR (126 MHz, acetone) δ 165.13, 158.06, 144.98, 141.98, 137.95, 135.46, 132.46, 131.72, 127.90, 126.89, 126.64, 126.17, 123.38, 121.88, 121.09. HRMS [M+H] + = 372.0027; [M+Na] + =393.9846.

[0395] Step 2: 4-(5-chlorothiophene-2-yl)-N-(pyridin-3-yl)benzothiazol-6-carboxamide HCl salt (Compound 13)

[0396] Compound 13 is prepared according to a similar method described in step 2 of the preparation of compound 11.

[0397] Preparation of 2-amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzo[d]thiazol-6-carboxamide HCl salt (compound 1)

[0398]

[0399] Step 1: Methyl 2-aminobenzo[d]thiazole-6-carboxylate

[0400] Bromine (3.10 g, 19.85 mmol) was dissolved in acetic acid (18 mL), and methyl 4-aminobenzoate (3.00 g, 19.85 mmol) and KSCN (6.89 g, 70.86 mmol) in acetic acid (67 mL) were slowly added dropwise. The reaction was allowed to proceed overnight at room temperature. The mixture was quenched with water, and the pH was adjusted to 9 with saturated Na₂CO₃ solution. The mixture was extracted three times with EA, washed with saturated brine, dried over anhydrous Na₂SO₄, and then filtered and purified by column chromatography (PE / EA = 3 / 1) to give the title compound (3.30 g, yield: 80%) as a white solid. LCMS: calculated exact mass = 208.0, found (ESI+) [M+H] + = 209.0.

[0401] Step 2: Methyl 2-amino-4-bromobenzo[d]thiazole-6-carboxylate

[0402] NBS (1.25 g, 7.21 mmol) was added to a solution of methyl 2-aminobenzo[d]thiazolyl-6-carboxylate (1 g, 4.81 mmol) in DMF (11 mL), and the mixture was stirred at 20 °C for 3 h. After the reaction was complete, the mixture was diluted with EA (20 mL) and washed with H2O (3 x 20 mL), aqueous Na2SO3 solution, and brine. The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid chromatography on silica gel (PE / EA = 3 / 1) to give the desired product as a yellow solid (1.0 g, 72.7% yield). LCMS: calculated exact mass = 285.9, found (ESI+) [M+H] + =286.9, 288.9.

[0403] Step 3: Methyl 4-bromo-2-((tert-butoxycarbonyl)amino)benzo[d]thiazol-6-carboxylate

[0404] DMAP (128 mg, 1.05 mmol) was added to a mixture of methyl 2-amino-4-bromobenzo[d]thiazolyl-6-carboxylate (1.5 g, 5.25 mmol), TEA (1 g, 10.49 mmol), and Boc2O (1.37 mg, 6.29 mmol) in DMF (14 mL). The mixture was stirred at 25 °C for 3 h. After the reaction was complete, the resulting reaction mixture was diluted with water (20 mL) and extracted with EA (3 × 30 mL). The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by rapid chromatography (PE:EA = 3:1) to give the desired product (1.2 g, 74.5% yield) as a white solid. LCMS: calculated exact mass = 386.0, found (ESI+) [M+H] + = 387.0, 389.0.

[0405] Step 4: 4-Bromo-2-((tert-butoxycarbonyl)amino)benzo[d]thiazolyl-6-carboxylic acid

[0406] LiOH (250 mg, 10.36 mmol) in H₂O (8 mL) was added to a solution of methyl 4-bromo-2-((tert-butoxycarbonyl)amino)benzo[d]thiazol-6-carboxylate (1.0 g, 2.59 mmol) in MeOH (40 mL). The reaction was stirred at 50 °C for 2 h, and an aqueous solution of HCl (1 N) was added to adjust the pH to < 3. The mixture was extracted with EA (3 × 20 mL), and the combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated under vacuum to give the desired product as a white solid (820 mg, 85.2% yield). LCMS: calculated exact mass = 372.0, found (ESI+) [M+H] + =373.0, 375.0.

[0407] Step 5: (4-bromo-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl)tert-butyl carbamate

[0408] EDCI (0.5 g, 2.64 mmol) was added to a mixture of 4-bromo-2-((tert-butoxycarbonyl)amino)benzo[d]thiazolyl-6-carboxylic acid (820 mg, 2.2 mmol), DMAP (220 mg, 2.46 mmol), and pyridine-3-amine (250 mg, 2.64 mmol) in DMF (6 mL), and the reaction was stirred at 20 °C for 16 h. The mixture was diluted with EA (30 mL) and washed with brine (5 x 30 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by chromatography on silica gel (DCM / MeOH = 10 / 1) to give the desired product as a white solid (0.8 g, 83.1% yield). LCMS: calculated exact mass = 448.0, found (ESI+) [M+H] + = 449.0, 451.0.

[0409] Step 6: (4-(3-chlorophenyl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl)tert-butyl carbamate

[0410] A mixture of tert-butyl (4-bromo-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)carbamate (0.4 g, 0.89 mmol), (3-chlorophenyl)boronic acid (153 mg, 4.31 mmol), Pd(dppf)Cl2 (80 mg, 0.11 mmol), and Cs2CO3 (0.48 g, 1.51 mmol) in 1,4-dioxane (12 mL) and H2O (2 mL) was stirred at 120 °C for 2 h under nitrogen. The resulting mixture was concentrated under vacuum, and the residue was purified by chromatography on silica gel (DCM / MeOH = 10 / 1) to give the desired product (260 mg, 60.7% yield) as a yellow solid. LCMS: calculated exact mass = 480.1, found (ESI+) [M+H] + = 481.1.

[0411] Step 7: 2-Amino-4-(3-chlorophenyl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0412] At 0 °C, 4 N HCl (in 10 mL of 1,4-dioxane) was added to a solution of (200 mg, 0.42 mmol) of tert-butyl carbamate in DCM (5 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was suspended in MTBE (20 mL) and stirred for 0.5 h, filtered, washed with MTBE, and the solid was collected and dried under vacuum to give the desired product (103 mg, 59.2% yield) as a white solid. LCMS: calculated exact mass = 380.0, found (ESI+) [M+H] + = 381.0.

[0413] 1 H NMR (400 MHz, DMSO-d6) δ 10.39 (s, 1H), 8.94 (d, J = 2.3 Hz, 1H), 8.34 (d, J = 1.8 Hz, 1H), 8.31 (dd, J = 4.7, 1.3 Hz, 1H), 8.22 – 8.17 (m,1H), 8.05 – 7.98 (m, 3H), 7.93 (t, J = 1.8 Hz, 1H), 7.83 – 7.79 (m, 1H), 7.52(t, J = 7.9 Hz, 1H), 7.44 (ddd, J = 8.0, 2.1, 1.1 Hz, 1H), 7.40 (dd, J = 8.2,4.6 Hz, 1H).

[0414] Preparation of 2-amino-4-(3,6-dihydro-2H-pyran-4-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (compound 14)

[0415] Step 1: (4-(3,6-dihydro-2H-pyran-4-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0416] To a mixture of (4-bromo-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl (200 mg, 0.445 mmol) in dioxane (4 mL), add H₂O (2 mL), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentane (105 mg, 0.49 mmol), Pd(dppf)Cl₂ (40 mg, 0.05 mmol), and Cs₂CO₃ (290 mg, 0.82 mmol). Stir the mixture at 120 °C for 2 h under nitrogen atmosphere. After completion, the resulting reaction mixture was diluted with H₂O, extracted with EA, washed with brine, dried over Na₂SO₄, filtered, and concentrated. The residue was purified by rapid chromatography using flowing MeOH (0-10%) in DCM to give the desired product as a yellow solid (150 mg, 74.4% yield). LCMS: calculated exact mass = 452.2, found (ESI+) [M+H]+ = 453.2.

[0417] Step 2: 2-Amino-4-(3,6-dihydro-2H-pyran-4-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0418] At 0 °C, 4 N HCl (in 1,4-dioxane, 10 mL) was added to a solution of (4-(3,6-dihydro-2H-pyran-4-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (100 mg, 0.445 mmol) in DCM (5 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (65 mg, 83.3% yield) as a white solid. LCMS: calculated exact mass = 352.1, found (ESI+) [M+H] + = 353.1.

[0419] 1H NMR (400 MHz, DMSO-d6) δ 10.34 (s, 1H), 8.93 (d, J = 2.3 Hz, 1H), 8.30 (dd, J = 4.7, 1.5 Hz, 1H), 8.23 ​​(d, J = 1.8 Hz, 1H), 8.21 – 8.15 (m,1H), 7.91 (s, 2H), 7.78 (d, J = 1.8 Hz, 1H), 7.39 (dd, J = 8.3, 4.7 Hz, 1H), 6.58 – 6.55 (m, 1H), 4.27 (m, 2H), 3.86 (t, J = 5.5 Hz, 2H), 2.71 – 2.65 (m,2H).

[0420] Preparation of 2-amino-N-(pyridin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)benzo[d]thiazolyl-6-carboxamide (compound 15)

[0421]

[0422] Step 1: (6-(pyridin-3-ylcarbamoyl)-4-(tetrahydro-2H-pyran-4-yl)benzo[d]thiazolyl)tert-butyl carbamate

[0423] Pd(OH)₂ / C (50 mg) was added to a solution of (200 mg, 0.412 mmol) tert-butyl carbamate (4-(3,6-dihydro-2H-pyran-4-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate in MeOH (10 mL). The mixture was hydrogenated at 40 °C under 45 psi H₂ for 5 h. After the reaction was complete, the reaction was filtered through diatomaceous earth and the filtrate was concentrated under reduced pressure to give the desired product as a yellow solid (163 mg, 81.1% yield). LCMS: calculated exact mass = 454.2, found (ESI+) [M+H] + = 455.2.

[0424] Step 2: 2-Amino-N-(pyridin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)benzo[d]thiazolyl-6-carboxamide

[0425] At 0 °C, 4 N HCl (in 1,4-dioxane, 10 mL) was added to a solution of (6-(pyridin-3-ylcarbamoyl)-4-(tetrahydro-2H-pyran-4-yl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (100 mg, 0.22 mmol) in DCM (5 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (60 mg, 75.1% yield) as a white solid. LCMS: calculated exact mass = 354.1, found (ESI+) [M+H] + = 355.1.

[0426] 1 H NMR (400 MHz, DMSO-d6) δ 10.28 (s, 1H), 8.92 (d, J = 2.4 Hz, 1H), 8.30 (dd, J = 4.7, 1.4 Hz, 1H), 8.22 – 8.16 (m, 2H), 7.90 (s, 2H), 7.78 (d, J = 1.6 Hz, 1H), 7.40 (dd, J = 8.3, 4.7 Hz, 1H), 4.00 (dd, J = 11.0, 3.6 Hz, 2H), 3.50 – 3.34 (m, 3H), 1.95 – 1.85(m, 2H), 1.78 – 1.75 (m, 2H).

[0427] Preparation of 2-amino-4-(4,4-difluorocyclohexyl)-N-(pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 16)

[0428]

[0429] Step 1: (4-(4,4-difluorocyclohexyl-1-en-1-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0430] To a mixture of (4-bromo-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl (300 mg, 0.67 mmol) in dioxane (6 mL), add H₂O (2 mL), 2-(4,4-difluorocyclohexyl-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentane (244 mg, 1.0 mmol), Pd(dppf)Cl₂ (73 mg, 0.1 mmol), and Cs₂CO₃ (650 mg, 2.0 mmol). Stir the mixture at 120 °C for 2 h under nitrogen atmosphere. After completion, the resulting reaction mixture was diluted with H₂O, extracted with EA, washed with brine, dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel chromatography using mobile MeOH (0-10%) in DCM to give the desired product as a yellow solid (240 mg, 73.8% yield). LCMS: calculated exact mass = 486.2, found (ESI+) [M+H]+ = 487.2.

[0431] Step 2: (4-(4,4-difluorocyclohexyl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl)tert-butyl carbamate

[0432] Pd / C (100 mg) and Pd(OH)₂ / C (50 mg) were added to a solution of (4-(4,4-difluorocyclohexyl-1-en-1-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)carbamate tert-butyl (100 mg, 0.20 mmol) in MeOH (4 mL) and THF (4 mL). The mixture was stirred at room temperature under 45 psi H₂ for 16 h. The reaction mixture was filtered and washed with MeOH, and the filtrate was concentrated to give the desired product (100 mg, 100% yield) as a pale yellow solid. LCMS: calculated exact mass = 488.2, found (ESI+) [M+H]+ = 489.2.

[0433] Step 3: 2-Amino-4-(4,4-difluorocyclohexyl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0434] To a solution of (100 mg, 0.20 mmol) tert-butyl carbamate in dioxane (3 mL), 4 NHCl and 0.2 mL of H₂O were added to dioxane (3 mL). The mixture was stirred at room temperature for 3 h. After completion, the resulting reaction was concentrated and purified by preparative HPLC to give the desired product (40 mg, 50.5% yield) as a white solid. LCMS: calculated exact mass = 388.1, found (ESI+) [M+H] + = 389.1.

[0435] 1 H NMR (400 MHz, CD3OD) δ 8.91 (d, J = 2.3 Hz, 1H), 8.33 – 8.26 (m,2H), 8.16 – 8.14 (m, 1H), 7.83 (s, 1H), 7.49 – 7.44 (m, 1H), 3.46 – 3.36 (m,1H), 2.27 – 2.15 (m, 2H), 2.12 – 1.87 (m, 6H).

[0436] Preparation of 4-(3,6-dihydro-2H-pyran-4-yl)-N-(pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 17) and N-(pyridin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)benzo[d]thiazol-6-carboxamide (compound 18)

[0437]

[0438]

[0439] Step 1: Methyl 4-bromobenzo[d]thiazole-6-carboxylate

[0440] t-BuONO (2.7 g, 26.13 mmol) was added to a solution of methyl 2-amino-4-bromobenzo[d]thiazolyl-6-carboxylate (2.5 g, 8.71 mmol) in THF (50 mL). The mixture was stirred under reflux at N2 for 2 h. After completion, the resulting reaction was concentrated under vacuum and purified by silica gel chromatography using mobile EA (0-10%) in PE to give the desired product as a white solid (1.3 g, 55.1% yield). LCMS: calculated exact mass = 270.9, found (ESI+) [M+H] + = 271.9, 273.9.

[0441] Step 2: 4-Bromobenzo[d]thiazolyl-6-carboxylic acid

[0442] LiOH (400 mg, 16.73 mmol) in H₂O (4 mL) was added to a solution of methyl 4-bromobenzo[d]thiazolyl-6-carboxylate (1.3 g, 4.78 mmol) in MeOH (20 mL). The reaction was stirred at 25 °C for 16 h, and an aqueous solution of HCl (1 N) was added to adjust the pH to < 3. The mixture was extracted with EA (3 × 20 mL), and the combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated under vacuum to give the desired product as a yellow solid (1.1 g, 90.2% yield). LCMS: calculated exact mass = 256.9, found (ESI+) [M+H] + = 257.9, 259.9.

[0443] Step 3: 4-Bromo-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0444] EDCI (0.53 g, 2.79 mmol) was added to a mixture of 4-bromobenzo[d]thiazolyl-6-carboxylic acid (600 mg, 2.33 mmol), DMAP (310 mg, 2.56 mmol), and pyridine-3-amine (260 mg, 2.79 mmol) in DMF (6 mL), and the reaction was stirred at 20 °C for 16 h. The mixture was diluted with EA (30 mL) and washed with brine (5 × 30 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by chromatography on silica gel (DCM / MeOH = 10 / 1) to give the desired product as a white solid (0.55 g, 70.87% yield). LCMS: calculated exact mass = 333.0, found (ESI+) [M+H] + = 334.0, 336.0.

[0445] Step 4: 4-(3,6-dihydro-2H-pyran-4-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0446] A mixture of 4-bromo-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (0.15 g, 0.45 mmol), Pd(dppf)Cl2 (73 mg, 0.10 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentane (120 mg, 0.58 mmol) and Cs2CO3 (0.29 g, 0.9 mmol) in 1,4-dioxane (2.5 mL) and H2O (0.5 mL) was stirred at 120 °C for 2 h under nitrogen. The resulting mixture was concentrated under vacuum, and the residue was purified by chromatography on silica gel (DCM / MeOH = 10 / 1) to give the desired product (120 mg, 78.95% yield) as a yellow solid. LCMS: Calculated exact mass = 337.1, measured value (ESI+) [M+H] + = 338.1.

[0447] 1 H NMR (400 MHz, CD3OD) δ 9.38 (s, 1H), 8.93 (d, J = 2.0 Hz, 1H), 8.62(d, J = 1.8 Hz, 1H), 8.34 – 8.28 (m, 2H), 8.03 (d, J = 1.8 Hz, 1H), 7.47(ddd, J = 8.4, 4.9, 0.6 Hz, 1H), 6.57– 6.54 (m, 1H), 4.40 (q, J = 2.8 Hz, 2H), 4.01 (t, J = 5.4 Hz, 2H), 2.86 – 2.81 (m, 2H).

[0448] Step 5: N-(pyridin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)benzo[d]thiazolyl-6-carboxamide

[0449] Pd / C (50 mg) and Pd(OH)₂ / C (50 mg) were added to a solution of 4-(3,6-dihydro-2H-pyran-4-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (120 mg, 0.36 mmol) in MeOH (4 mL) and THF (4 mL). The mixture was stirred at 60 °C for 16 h at 45 psi H₂. The reaction mixture was filtered and washed with MeOH, and the filtrate was concentrated and purified by preparative HPLC to give the desired product (50 mg, 41.67% yield) as a pale white solid. LCMS: calculated exact mass = 339.1, found (ESI+) [M+H]+ = 340.1.

[0450] 1 H NMR (400 MHz, CD3OD) δ 9.42 (s, 1H), 8.95 (d, J = 2.3 Hz, 1H), 8.61 (d, J = 1.7 Hz, 1H), 8.36 – 8.30 (m, 2H), 8.04 (d, J = 1.5 Hz, 1H), 7.50 (dd,J = 8.2, 4.7 Hz, 1H), 3.81 – 3.71 (m, 1H), 2.29 – 2.20 (m, 2H), 2.17 – 2.02(m, 6H).

[0451] Preparation of 4-(4,4-difluorocyclohexyl)-N-(pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 19)

[0452]

[0453] Step 1: 4-(4,4-difluorocyclohexyl-1-en-1-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0454] A mixture of 4-bromo-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (0.15 g, 0.45 mmol), Pd(dppf)Cl2 (73 mg, 0.10 mmol), 2-(4,4-difluorocyclohexyl-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentane (140 mg, 0.58 mmol) and Cs2CO3 (0.29 g, 0.9 mmol) in 1,4-dioxane (2.5 mL) and H2O (0.5 mL) was stirred at 120 °C for 2 h under nitrogen. The resulting mixture was concentrated under vacuum, and the residue was purified by chromatography on silica gel (DCM / MeOH = 10 / 1) to give the desired product (120 mg, 71.86% yield) as a yellow solid. LCMS: Calculated exact mass = 371.1, measured value (ESI+) [M+H] + = 372.1.

[0455] Step 2: 4-(4,4-difluorocyclohexyl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0456] Pd / C (150 mg) and Pd(OH)₂ / C (150 mg) were added to a solution of 4-(4,4-difluorocyclohexyl-1-en-1-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (100 mg, 0.27 mmol) in MeOH (3 mL) and THF (2 mL). The mixture was stirred at 50 °C for 16 h at 45 psi H₂. The reaction mixture was filtered and washed with MeOH, and the filtrate was concentrated and purified by preparative HPLC to give the desired product as a white solid (40 mg, 39.8% yield). LCMS: calculated exact mass = 373.1, found (ESI+) [M+H]+ = 374.1.

[0457] 1 H NMR (400 MHz, CD3OD) δ 9.42 (s, 1H), 8.95 (d, J = 2.3 Hz, 1H), 8.61 (d, J = 1.7 Hz, 1H), 8.36 – 8.30 (m, 2H), 8.04 (d, J = 1.5 Hz, 1H), 7.50 (dd,J = 8.2, 4.7 Hz, 1H), 3.82 – 3.70 (m, 1H), 2.29 – 2.20 (m, 2H), 2.17 – 2.02(m, 6H).

[0458] Preparation of 2-amino-4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-N-(pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 20)

[0459]

[0460] Step 1: (4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0461] To a mixture of (4-bromo-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl (200 mg, 0.445 mmol) in dioxane (4 mL), add H₂O (2 mL), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1,2,3,6-tetrahydropyridine (120 mg, 0.53 mmol), Pd(dppf)Cl₂ (40 mg, 0.05 mmol), and Cs₂CO₃ (290 mg, 0.82 mmol). Stir the mixture at 120 °C for 2 h under nitrogen atmosphere. After completion, the resulting reaction mixture was diluted with H₂O, extracted with EA, washed with brine, dried over Na₂SO₄, filtered, and concentrated. The residue was purified by rapid chromatography using flowing MeOH (0-10%) in DCM to give the desired product as a yellow solid (103 mg, 49.8% yield). LCMS: calculated exact mass = 465.2, measured (ESI+) [M+H]+ = 466.2.

[0462] Step 2: 2-Amino-4-(1-Methyl-1,2,3,6-tetrahydropyridin-4-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0463] At 0 °C, 4 NHCl (in 1,4-dioxane, 10 mL) was added to a solution of tert-butyl carbamate (100 mg, 0.215 mmol) in DCM (5 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (43 mg, 54.80% yield) as a white solid. LCMS: calculated exact mass = 365.1, found (ESI+) [M+H] + = 366.1.

[0464] 1 H NMR (400 MHz, DMSO-d6) δ 10.33 (s, 1H), 8.93 (d, J = 2.2 Hz, 1H), 8.30 (dd, J = 4.7, 1.4 Hz, 1H), 8.24 – 8.16 (m, 2H), 7.88 (s, 2H), 7.76 (d, J= 1.8 Hz, 1H), 7.39 (dd, J = 8.2, 4.7 Hz, 1H), 6.43 – 6.39 (m, 1H), 3.07–3.01 (m, 2H), 2.74– 2.67 (m, 2H), 2.58 (t, J = 5.6 Hz, 2H), 2.30 – 2.2 (m,2H).

[0465] Preparation of 2-amino-4-(1-methylpiperidin-4-yl)-N-(pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 21)

[0466]

[0467] Step 1: (4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0468] Pd(OH)₂ / C (50 mg) was added to a solution of tert-butyl carbamate (200 mg, 0.430 mmol) in MeOH (10 mL). The mixture was stirred at 40 °C for 16 h at 45 psi H₂. The reaction mixture was filtered and washed with MeOH, and the filtrate was concentrated and purified by preparative HPLC to give the desired product as a white solid (103 mg, 50.3% yield).

[0469] LCMS: Calculated exact mass = 467.2, measured value (ESI+) [M+H] + = 468.2.

[0470] Step 2: 2-Amino-4-(1-methylpiperidin-4-yl)-N-(pyridin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0471] At 0 °C, 4 NHCl (in 1,4-dioxane, 8 mL) was added to a solution of (100 mg, 0.213 mmol) of tert-butyl carbamate (4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate in DCM (2 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (34 mg, 43.26% yield) as a white solid. LCMS: calculated exact mass = 367.1, found (ESI+) [M+H] + = 368.1.

[0472] 1 H NMR (400 MHz, DMSO-d6) δ 10.29 (s, 1H), 8.92 (d, J = 2.1 Hz, 1H), 8.32 – 8.28 (m, 1H), 8.21– 8.15 (m, 2H), 7.90 – 7.74 (m, 3H), 7.39 (dd, J =8.3, 4.7 Hz, 1H), 3.21 – 3.11 (m, 1H), 2.94 – 2.86 (m, 2H), 2.22 (s, 3H), 2.02 – 1.74 (m, 6H).

[0473] Preparation of 2-amino-N-(pyridin-3-yl)-4-(tetrahydro-2H-pyran-3-yl)benzo[d]thiazolyl-6-carboxamide (compound 22)

[0474]

[0475] Step 1: (4-(3,4-dihydro-2H-pyran-5-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0476] To a mixture of (4-bromo-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl (200 mg, 0.445 mmol) in dioxane (4 mL), add H₂O (2 mL), 2-(3,4-dihydro-2H-pyran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentane (105 mg, 0.49 mmol), Pd(dppf)Cl₂ (40 mg, 0.05 mmol), and Cs₂CO₃ (290 mg, 0.82 mmol). Stir the mixture at 120 °C for 2 h under nitrogen atmosphere. After completion, the resulting reaction mixture was diluted with H₂O, extracted with EA, washed with brine, dried over Na₂SO₄, filtered, and concentrated. The residue was purified by rapid chromatography using flowing MeOH (0-10%) in DCM to give the desired product as a yellow solid (168 mg, 83.13% yield). LCMS: calculated exact mass = 452.2, found (ESI+) [M+H]+ = 453.2.

[0477] Step 2: (6-(pyridin-3-ylcarbamoyl)-4-(tetrahydro-2H-pyran-3-yl)benzo[d]thiazolyl)tert-butyl carbamate

[0478] Pd / C (50 mg) was added to a solution of tert-butyl (4-(3,4-dihydro-2H-pyran-5-yl)-6-(pyridin-3-ylcarbamoyl)benzo[d]thiazo-2-yl)carbamate (200 mg, 0.412 mmol) in MeOH (10 mL). The mixture was stirred at 40 °C for 16 h under 45 psi H2. The reaction mixture was filtered and washed with MeOH, and the filtrate was concentrated to give the desired product as a white solid (163 mg, 81.1% yield). LCMS: calculated exact mass = 454.2, found (ESI+) [M+H] + = 455.2.

[0479] Step 3: 2-Amino-N-(pyridin-3-yl)-4-(tetrahydro-2H-pyran-3-yl)benzo[d]thiazolyl-6-carboxamide

[0480] At 0 °C, 4 N HCl (in 10 mL of 1,4-dioxane) was added to a solution of (6-(pyridin-3-ylcarbamoyl)-4-(tetrahydro-2H-pyran-3-yl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (100 mg, 0.22 mmol) in DCM (2 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (66 mg, 86.44% yield) as a white solid. LCMS: calculated exact mass = 354.1, found (ESI+) [M+H] + = 355.1.

[0481] 1 H NMR (400 MHz, DMSO-d6) δ 10.30 (s, 1H), 8.92 (d, J = 2.4 Hz, 1H), 8.30 (dd, J = 4.7, 1.4 Hz, 1H), 8.21 – 8.15 (m, 2H), 7.92 (s, 2H), 7.74 (d, J= 1.5 Hz, 1H), 7.40 (dd, J = 8.3, 4.7 Hz, 1H), 3.97 – 3.86 (m, 2H), 3.54 –3.47 (m, 2H), 2.04 – 1.91(m, 3H), 1.75 – 1.64 (m, 2H).

[0482] Preparation of 2-amino-4-(3-chlorophenyl)-N-(pyridazin-3-yl)benzo[d]thiazol-6-carboxamide (compound 23)

[0483]

[0484] Step 1: 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylic acid ethyl ester

[0485] H₂O (8 mL), (3-chlorophenyl)boronic acid (2 g, 13 mmol), Pd(dppf)Cl₂ (732 mg, 1 mmol), and Cs₂CO₃ (6.5 g, 20 mmol) were added to a mixture of ethyl 4-bromo-2-((tert-butoxycarbonyl)amino)benzo[d]thiazolium-6-carboxylate (4 g, 10 mmol) in 1,4-dioxane (40 mL). The mixture was stirred at 120 °C for 2 h under nitrogen. After completion, the resulting reaction mixture was diluted with H₂O, extracted with EA, washed with brine, dried over Na₂SO₄, filtered, and concentrated. The residue was purified by rapid chromatography using mobile EA (0–10%) in PE to give the desired product (3.6 g, 83.72% yield) as a yellow solid. LCMS: Calculated exact mass = 432.1, measured value (ESI+) [M+H]+ = 433.1.

[0486] Step 2: 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylic acid

[0487] LiOH (870 mg, 36.4 mmol) in H₂O (13 mL) was added to a solution of ethyl 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylate (4.5 g, 10.4 mmol) in MeOH (40 mL). The reaction was stirred at 40 °C for 16 h, and an aqueous solution of HCl (1 N) was added to adjust the pH to < 3. The mixture was extracted with EA (3 × 20 mL), and the combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated under vacuum to give the desired product (4.1 g, 97.6% yield) as a white solid. LCMS: calculated exact mass = 404.1, found (ESI+) [M+H] + = 405.1.

[0488] Step 3: (4-(3-chlorophenyl)-6-(pyridazin-3-ylcarbamoyl)benzo[d]thiazolyl)tert-butyl carbamate

[0489] At 0 °C, DIEA (111 mg, 0.86 mmol), HATU (122 mg, 0.32 mmol), and pyridazine-3-amine (31 mg, 0.32 mmol) were added sequentially to a solution of 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (100 mg, 0.25 mmol) in THF (10 mL). The mixture was then stirred at 25 °C for 16 h. After the reaction was complete, the resulting mixture was diluted with H2O (50 mL) and extracted with EA (3 × 20 mL). The organic layer was washed with brine (3 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid chromatography on silica gel (DCM:MeOH = 20:1) to give the desired product as a solid (70 mg, 58.33% yield). LCMS: Calculated exact mass = 481.1, measured value (ESI+) [M+H] + = 482.1.

[0490] Step 4: 2-Amino-4-(3-chlorophenyl)-N-(pyridazin-3-yl)benzo[d]thiazolyl-6-carboxamide

[0491] At 0 °C, 4 N HCl (in 1,4-dioxane, 10 mL) was added to a solution of (200 mg, 0.42 mmol) of tert-butyl carbamate in DCM (4 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (100 mg, 62.5% yield) as a white solid. LCMS: calculated exact mass = 381.0, found (ESI+) [M+H] + = 382.0.

[0492] 1H NMR (400 MHz, DMSO-d6) δ 11.51 (s, 1H), 9.02 (d, J = 4.6 Hz, 1H), 8.44 – 8.38 (m, 2H), 8.15 (d, J = 1.2 Hz, 1H), 8.10 – 7.98 (m, 3H), 7.91 (d,J = 7.6 Hz, 1H), 7.74 (dd,J = 9.0, 4.7 Hz, 1H), 7.54 – 7.41 (m, 2H).

[0493] Preparation of 2-amino-4-(3-chlorophenyl)-N-(pyrimidin-5-yl)benzo[d]thiazol-6-carboxamide (compound 24)

[0494]

[0495] Step 1: (4-(3-chlorophenyl)-6-(pyrimidin-5-ylcarbamoyl)benzo[d]thiazolyl)tert-butyl carbamate

[0496] At 0 °C, DIEA (64 mg, 0.50 mmol), HATU (95 mg, 0.30 mmol), and pyrimidine-5-amine (24 mg, 0.25 mmol) were added sequentially to a solution of 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylic acid (100 mg, 0.25 mmol) in DMF (3 mL). The mixture was then stirred at 25 °C for 16 h. After the reaction was complete, the resulting mixture was diluted with H2O (50 mL) and extracted with EA (3 x 20 mL). The organic layer was washed with brine (3 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid chromatography on silica gel (DCM:MeOH = 20:1) to give the desired product as a solid (20 mg, 16.7% yield). LCMS: Calculated exact mass = 481.1, measured value (ESI+) [M+H] + = 482.1.

[0497] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(pyrimidin-5-yl)benzo[d]thiazolyl-6-carboxamide

[0498] At 0 °C, 4 N HCl (in 1,4-dioxane, 5 mL) was added to a solution of (50 mg, 0.10 mmol) of tert-butyl carbamate (4 mL) in DCM, and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (30 mg, 78.9% yield) as a white solid. LCMS: calculated exact mass = 381.0, found (ESI+) [M+H] + = 382.0.

[0499] 1 H NMR (400 MHz, CD3OD) δ 9.22 (s, 2H), 8.90 (s, 1H), 8.29 (d, J = 1.9Hz, 1H), 8.02 (d, J = 1.9 Hz, 1H), 7.91 (t, J = 1.8 Hz, 1H), 7.77 – 7.73 (m,1H), 7.45 (t, J = 7.9 Hz, 1H), 7.40 – 7.36 (m, 1H).

[0500] Preparation of 2-amino-N-(2-aminopyrimidin-5-yl)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxamide (compound 25)

[0501]

[0502] Step 1: (6-((2-aminopyrimidin-5-yl)carbamoyl)-4-(3-chlorophenyl)benzo[d]thiazolyl)tert-butyl carbamate

[0503] At 0 °C, DIEA (390 mg, 3.0 mmol), HATU (285 mg, 0.75 mmol), and pyrimidine-2,5-diamine (100 mg, 0.90 mmol) were added sequentially to a solution of 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylic acid (300 mg, 0.75 mmol) in DMF (20 mL). The mixture was then stirred at 25 °C for 16 h. After the reaction was complete, the resulting mixture was diluted with H2O (50 mL) and extracted with EA (3 × 20 mL). The organic layer was washed with brine (3 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid chromatography on silica gel (DCM:MeOH = 20:1) to give the desired product as a solid (250 mg, 67.2% yield). LCMS: Calculated exact mass = 496.1, measured value (ESI+) [M+H] + = 497.1.

[0504] Step 2: 2-Amino-N-(2-aminopyrimidin-5-yl)-4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxamide

[0505] At 0 °C, 4 N HCl (in 1,4-dioxane, 10 mL) was added to a solution of (250 mg, 0.5 mmol) of tert-butyl carbamate in DCM (4 mL), and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product as a white solid (120 mg, 60.6% yield). LCMS: calculated exact mass = 396.1, found (ESI+) [M+H] + = 397.1.

[0506] 1H NMR (400 MHz, DMSO-d6) δ 10.06 (s, 1H), 8.51 (s, 2H), 8.29 (d, J =1.7 Hz, 1H), 8.02 – 7.97 (m, 3H), 7.92 (d, J = 1.8 Hz, 1H), 7.82 – 7.78 (m,1H), 7.51 (t, J = 7.8 Hz, 1H), 7.47 – 7.42 (m, 1H), 6.53 (s, 2H).

[0507] Preparation of 2-amino-4-(3-chlorophenyl)-N-(1-methyl-1H-imidazol-4-yl)benzo[d]thiazol-6-carboxamide (compound 26)

[0508]

[0509] Step 1: (4-(3-chlorophenyl)-6-((1-methyl-1H-imidazol-4-yl)carbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0510] At 0 °C, DIEA (390 mg, 3.0 mmol), HATU (285 mg, 0.75 mmol), and 1-methyl-1H-imidazol-4-amine (87 mg, 0.90 mmol) were added sequentially to a solution of 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylic acid (300 mg, 0.75 mmol) in DMF (10 mL). The mixture was then stirred at 25 °C for 16 h. After the reaction was complete, the resulting mixture was diluted with H2O (50 mL) and extracted with EA (3 × 20 mL). The organic layer was washed with brine (3 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid chromatography on silica gel (DCM:MeOH = 20:1) to give the desired product as a solid (100 mg, 27.5% yield). LCMS: Calculated exact mass = 483.1, measured value (ESI+) [M+H] + = 484.1.

[0511] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(1-methyl-1H-imidazol-4-yl)benzo[d]thiazol-6-carboxamide

[0512] At 0 °C, 4 N HCl (in 1,4-dioxane, 10 mL) was added to a solution of (100 mg, 0.21 mmol) of tert-butyl carbamate (4 mL) in DCM, and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (40 mg, 49.4% yield) as a white solid. LCMS: calculated exact mass = 383.1, found (ESI+) [M+H] + = 384.1.

[0513] 1 H NMR (400 MHz, DMSO-d6) δ 10.78 (s, 1H), 8.31 (d, J = 1.8 Hz, 1H), 8.07 (d, J = 1.8 Hz, 1H), 8.00 (t, J = 1.8 Hz, 1H), 7.94 (s, 2H), 7.91 – 7.87(m, 1H), 7.49 (t, J = 7.9 Hz, 1H), 7.46 – 7.40 (m, 2H), 7.38 (d, J = 1.3 Hz, 1H), 3.66 (s, 3H).

[0514] Preparation of 2-amino-4-(3-chlorophenyl)-N-(1-methyl-1H-pyrazol-4-yl)benzo[d]thiazole-6-carboxamide (compound 28)

[0515]

[0516] Step 1: (4-(3-chlorophenyl)-6-((1-methyl-1H-pyrazole-4-yl)carbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0517] At 0 °C, DIEA (390 mg, 3.0 mmol), HATU (285 mg, 0.75 mmol), and 1-methyl-1H-pyrazole-4-amine (87 mg, 0.90 mmol) were added sequentially to a solution of 2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylic acid (300 mg, 0.75 mmol) in DMF (10 mL). The mixture was then stirred at 25 °C for 16 h. After the reaction was complete, the resulting mixture was diluted with H2O (50 mL) and extracted with EA (3 × 20 mL). The organic layer was washed with brine (3 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid chromatography on silica gel (DCM:MeOH = 20:1) to give the desired product as a solid (250 mg, 68.9% yield). LCMS: Calculated exact mass = 483.1, measured value (ESI+) [M+H] + = 484.1.

[0518] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(1-methyl-1H-pyrazol-4-yl)benzo[d]thiazol-6-carboxamide

[0519] At 0 °C, 4 N HCl (in 1,4-dioxane, 10 mL) was added to a solution of (250 mg, 0.52 mmol) of tert-butyl carbamate (4 mL) in DCM, and the reaction was stirred at 25 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in EA (20 mL), washed with an aqueous solution of NaHCO3 (20 mL), brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (125 mg, 63.1% yield) as a white solid. LCMS: calculated exact mass = 383.1, found (ESI+) [M+H] + = 384.1.

[0520] 1H NMR (400 MHz, CD3OD)δ 8.20 (d, J = 1.0 Hz, 1H), 8.03 (s, 1H), 7.93 (d, J = 1.3 Hz, 1H), 7.89 – 7.87 (m, 1H), 7.76 – 7.71 (m, 1H), 7.65 (s, 1H), 7.44 (t, J = 7.8 Hz, 1H), 7.38 – 7.35 (m, 1H), 3.89 (s, 3H).

[0521] Preparation of 2-amino-N-(6-aminopyridin-3-yl)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxamide (compound 29)

[0522]

[0523] Step 1: (6-((6-aminopyridin-3-yl)carbamoyl)-4-(3-chlorophenyl)benzo[d]thiazolyl)tert-butyl carbamate

[0524]

[0525] To a solution of 2-((tert-butoxycarbonyl)amino)-4-(2-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (500 mg, 1.2 mmol) in DMF (10 mL), pyridine-2,5-diamine (202 mg, 1.8 mmol), HATU (940 mg, 2.5 mmol), and DIEA (399 mg, 3.0 mmol) were added. The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (30 mL × 3). The organic layers were combined, washed with brine (20 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to give the product as a brown solid (500 mg, 82% yield). LCMS: calculated exact mass = 495.1, found (ESI+) [M+H] + = 496.1.

[0526] Step 2: 2-Amino-N-(6-aminopyridin-3-yl)-4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxamide

[0527]

[0528] A solution of 4 M HCl in dioxane (10 mL) was added to a solution of (6-((6-aminopyridin-3-yl)carbamoyl)-4-(3-chlorophenyl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (500 mg, 1.0 mmol) in DCM (10 mL). The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a N2 atmosphere. The reaction was monitored by LCMS. After completion, the solid was collected by filtration, washed with DCM (30 mL × 3), and dried under reduced pressure. The crude product was purified by MPLC (CH3OH / H2O) to give the product as a gray solid (20 mg, 5% yield). LCMS: calculated exact mass = 395.9, found (ESI+) [M+H] + = 396.9.

[0529] 1 H NMR (400 MHz, DMSO) δ 9.97 (s, 1H), 8.28 (d, J = 1.6 Hz, 1H), 8.22 (d, J = 2.4 Hz, 1H), 7.97 (s, 3H), 7.92 (t, J = 1.7 Hz, 1H), 7.81 (d, J = 7.8Hz, 1H), 7.71 (dd, J = 8.8, 2.6 Hz, 1H), 7.51 (t, J = 7.8 Hz, 1H), 7.46 –7.41 (m, 1H), 6.47 (d, J = 8.8 Hz, 1H), 5.81 (s, 2H).

[0530] Preparation of N-(6-aminopyridin-3-yl)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxamide (compound 30)

[0531]

[0532]

[0533] Step 1: Ethyl 4-bromobenzo[d]thiazole-6-carboxylate

[0534]

[0535] Amyl nitrite (9.4 g, 79.8 mmol) was added to a solution of ethyl 2-amino-4-bromobenzo[d]thiazolyl-6-carboxylate (12 g, 39.9 mmol) in THF (120 mL). The reaction mixture was stirred in an oil bath at 70 °C for 0.5 h via a balloon under N2 atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (100 mL) and extracted with EA (100 mL × 3). The organic layers were combined and washed with brine (100 mL × 3) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by silica gel rapid chromatography (PE / EA = 3 / 1) to give the product as a yellow solid (4.5 g, 39% yield). LCMS: calculated exact mass = 285.0, found (ESI+) [M+H] + = 286.0.

[0536] Step 2: Ethyl 4-(3-chlorophenyl)benzo[d]thiazol-6-carboxylate

[0537]

[0538] At 25 °C, (3-chlorophenyl)boronic acid (2.4 g, 15.4 mmol), Pd(dppf)Cl2 (562 mg, 0.8 mmol), and K2CO3 (2.1 g, 15.4 mmol) were added to a solution of ethyl 4-bromobenzo[d]thiazolyl-6-carboxylate (2.2 g, 7.7 mmol) in 1,4-dioxane (12.5 mL) and H2O (2.5 mL). The reaction mixture was stirred at 120 °C for 2 h under microwave irradiation. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (40 mL × 3). The organic layers were combined, washed with brine (30 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by silica gel rapid chromatography (PE / EA = 3 / 1) to give the product as a white solid (1.1 g, 46% yield). LCMS: Calculated exact mass = 317.0, measured value (ESI+) [M+H] + = 318.0.

[0539] Step 3: 4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid

[0540]

[0541] LiOH (332 mg, 13.8 mmol) was added to a solution of ethyl 4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylate (1.1 g, 3.5 mmol) in MeOH (10 mL) and H₂O (2.5 mL). The reaction mixture was stirred in an oil bath at 60 °C for 5 h by a balloon under N₂ atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was adjusted to pH 3 with HCl (3 mol / L). The reaction mixture was quenched with water (20 mL) and extracted with EA (20 mL × 3). The organic layers were combined and washed with brine (20 mL × 3) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to give the product as a white solid (900 mg, 90% yield). LCMS: calculated exact mass = 289.0, found (ESI+) [MH] + = 288.0.

[0542] Step 4: Synthesis of N-(6-aminopyridin-3-yl)-4-(3-chlorophenyl)benzo[d]thiazol-6-carboxamide (compound 30)

[0543]

[0544] To a solution of 4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (200 mg, 0.7 mmol) in DMF (10 mL), pyridine-2,5-diamine (152 mg, 1.4 mmol), HATU (532 mg, 1.4 mmol), and DIEA (270 mg, 2.1 mmol) were added. The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (30 mL) and extracted with EA (50 mL × 3). The organic layers were combined, washed with brine (20 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by MPLC (MeOH / H2O) to give the product as a purple solid (37.7 mg, 14% yield). LCMS: calculated exact mass = 380.0, found (ESI+) [M+H] + = 381.0.

[0545] 1H NMR (400 MHz, DMSO) δ 10.91 (s, 1H), 9.65 (s, 1H), 8.87 (d, J =1.5 Hz, 1H), 8.57 (d, J = 2.0 Hz, 1H), 8.28 (d, J = 1.5 Hz, 1H), 8.23 ​​(dd, J= 9.5, 2.1 Hz, 1H), 8.05 (s, 1H), 7.94 (d, J = 7.5 Hz, 2H), 7.64 – 7.51 (m,2H), 7.06 (d, J = 9.5 Hz, 1H).

[0546] Preparation of 4-(3-chlorophenyl)-N-(6-morpholinopyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 31)

[0547]

[0548] To a solution of 4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (400 mg, 1.4 mmol) in DMF (10 mL), 6-morpholinopyridine-3-amine (500 mg, 2.8 mmol), HATU (760 mg, 2.0 mmol), and DIEA (387 mg, 3.0 mmol) were added. The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (30 mL) and extracted with EA (50 mL × 3). The organic layers were combined, washed with brine (20 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by MPLC (MeOH / H2O) to give the product as a purple solid (235 mg, 37% yield). LCMS: calculated exact mass = 450.1, found (ESI+) [M+H] + = 451.2.

[0549] 1H NMR (400 MHz, MeOD) δ 9.44 (s, 1H), 8.81 (d, J = 2.4 Hz, 1H), 8.73 (d, J = 1.7 Hz, 1H), 8.29 (dd, J = 9.8, 2.5 Hz, 1H), 8.22 (d, J = 1.7 Hz,1H), 7.97 (t, J = 1.7 Hz, 1H), 7.89 – 7.81 (m, 1H), 7.56 – 7.44 (m, 3H), 3.94 – 3.83 (m, 4H), 3.75 – 3.63 (m, 4H).

[0550] Preparation of 4-(3-chlorophenyl)-N-(6-(piperazin-1-yl)pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 32)

[0551]

[0552] Step 1: 4-(5-(4-(3-chlorophenyl)benzo[d]thiazolyl-6-carbamate)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester

[0553]

[0554] To a solution of 4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (300 mg, 1.0 mmol) in DMF (10 mL), tert-butyl 4-(5-aminopyridin-2-yl)piperazine-1-carboxylic acid (500 mg, 1.8 mmol), HATU (760 mg, 2.0 mmol), and DIEA (387 mg, 3.0 mmol) were added. The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (20 mL × 3). The organic layers were combined and washed with brine (20 mL × 3) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by silica gel rapid chromatography (PE / EA = 4 / 1) to give the product as a yellow solid (300 mg, 52% yield). LCMS: Calculated exact mass = 549.2, measured value (ESI+) [M+H] + = 550.2.

[0555] Step 2: 4-(3-chlorophenyl)-N-(6-(piperazin-1-yl)pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 32)

[0556]

[0557] A solution of 4M HCl in dioxane (10 mL) was added to a solution of 200 mg (0.36 mmol) of tert-butyl 4-(5-(4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxamido)pyridin-2-yl)piperazine-1-carboxylate in DCM (10 mL). The mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a N2 atmosphere. The reaction was monitored by LCMS. After completion, the solid was collected by filtration, washed with DCM (30 mL × 3), and dried under reduced pressure to give the product HCl salt as a yellow solid (95 mg, 54% yield). LCMS: calculated exact mass = 449.1, found (ESI+) [M+H] + = 450.0.

[0558] 1 H NMR (400 MHz, MeOD) δ 9.45 (s, 1H), 8.91 (d, J = 2.5 Hz, 1H), 8.75 (d, J = 1.7 Hz, 1H), 8.40 (dd, J = 9.8, 2.5 Hz, 1H), 8.23 ​​(d, J = 1.8 Hz,1H), 7.97 (t, J = 1.7 Hz, 1H), 7.84 (dt, J = 7.6, 1.4 Hz, 1H), 7.59 – 7.53(m, 1H), 7.53 – 7.45 (m, 2H), 4.06 – 3.96 (m, 4H), 3.54 – 3.44 (m, 4H).

[0559] Preparation of 2-amino-4-(3-chlorophenyl)-N-(6-morpholinopyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (compound 33)

[0560]

[0561] Step 1: (4-(3-chlorophenyl)-6-((6-morpholinopyridin-3-yl)carbamoyl)benzo[d]thiazolyl-2-yl)tert-butyl carbamate

[0562]

[0563] To a solution of 2-((tert-butoxycarbonyl)amino)-4-(2-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (500 mg, 1.2 mmol) in DMF (10 mL), 6-morpholinopyridine-3-amine (332 mg, 1.8 mmol), HATU (940 mg, 2.5 mmol), and DIEA (399 mg, 3.0 mmol) were added. The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (30 mL × 3). The organic layers were combined, washed with brine (20 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to give the product as a brown solid (500 mg, 72% yield). LCMS: calculated exact mass = 565.2, found (ESI+) [M+H] + = 566.2.

[0564] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(6-morpholinopyridin-3-yl)benzo[d]thiazolyl-6-carboxamide (Compound 33)

[0565]

[0566] A solution of 4M HCl in dioxane (10 mL) was added to a solution of (4-(3-chlorophenyl)-6-((6-morpholinopyridin-3-yl)carbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (500 mg, 0.9 mmol) in DCM (10 mL). The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under N2 atmosphere. The reaction was monitored by LCMS. After completion, the solid was collected by filtration, washed with DCM (30 mL × 3) and dried under reduced pressure. The crude product was purified by MPLC (CH3OH / H2O, containing 0.1% HCl) to give the product as a yellow solid (90 mg, crude product). The crude product was further purified by preparative HPLC (CH3CN / H2O, containing 0.05% FA) to give the product as a pale yellow solid (34.1 mg, 29% yield). LCMS: Calculated exact mass = 466.0, measured value (ESI+) [M+H] + = 467.0.

[0567] 1H NMR (400 MHz, DMSO) δ 10.12 (s, 1H), 8.48 (d, J = 2.6 Hz, 1H), 8.30 (d, J = 1.8 Hz, 1H), 7.98 (d, J = 1.7 Hz, 3H), 7.96 – 7.91 (m, 2H), 7.83– 7.77 (m, 1H), 7.51 (t, J = 7.9 Hz, 1H), 7.43 (ddd, J = 8.0, 2.1, 1.1 Hz,1H), 6.87 (d, J = 9.1 Hz, 1H), 3.74 – 3.69 (m, 4H), 3.42 – 3.38 (m, 4H).

[0568] Preparation of 2-amino-4-(3-chlorophenyl)-N-(6-(piperazin-1-yl)pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 34)

[0569]

[0570] Step 1: 4-(5-(2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxamido)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester

[0571]

[0572] To a solution of 2-((tert-butoxycarbonyl)amino)-4-(2-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (500 mg, 1.2 mmol) in DMF (10 mL), tert-butyl 4-(5-aminopyridin-2-yl)piperazine-1-carboxylic acid (516 mg, 1.8 mmol), HATU (940 mg, 2.5 mmol), and DIEA (399 mg, 3.0 mmol) were added. The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (30 mL × 3). The organic layers were combined, washed with brine (20 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by silica gel rapid chromatography (PE / EA = 4 / 1) to give the product as a yellow solid (380 mg, 61% yield). LCMS: Calculated exact mass = 664.2, measured value (ESI+) [M+H] + = 665.2.

[0573] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(6-(piperazin-1-yl)pyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 34)

[0574]

[0575] A solution of 4M HCl in dioxane (10 mL) was added to a solution of 4-(5-(2-((tert-butoxycarbonyl)amino)-4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxamido)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (380 mg, 0.6 mmol) in DCM (10 mL). The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a N2 atmosphere. The reaction was monitored by LCMS. After completion, the solid was collected by filtration, washed with DCM (30 mL × 3), and dried under reduced pressure to give the product HCl salt (267 mg, 93% yield) as a pale yellow solid. LCMS: calculated exact mass = 501.4, found (ESI+) [M+H] + = 502.4.

[0576] 1 H NMR (400 MHz, DMSO) δ 10.08 (s, 1H), 8.44 (d, J = 2.6 Hz, 1H), 8.30 (d, J = 1.8 Hz, 1H), 7.98 (s, 3H), 7.93 – 7.87 (m, 2H), 7.83 – 7.78 (m,1H), 7.51 (t, J = 7.9 Hz, 1H), 7.44 (ddd, J = 8.0, 2.1, 1.1 Hz, 1H), 6.83 (d,J = 9.2 Hz, 1H), 3.82 (s, 1H), 3.38 – 3.35 (m, 4H), 2.88 – 2.76 (m, 4H).

[0577] Preparation of 2-amino-4-(3-chlorophenyl)-N-(6-hydroxypyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 35)

[0578]

[0579] Step 1: (4-(3-chlorophenyl)-6-((6-hydroxypyridin-3-yl)carbamoyl)benzo[d]thiazolyl)tert-butyl carbamate

[0580]

[0581] Add 5-aminopyridine-2(1H)-one (204 mg, 1.8 mmol), BTFFH (782 mg, 2.5 mmol), and DIEA (399 mg, 3.0 mmol) to a solution of 2-((tert-butoxycarbonyl)amino)-4-(2-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (500 mg, 1.2 mmol) in DMF (10 mL) to a solution of 2-((tert-butoxycarbonyl)amino)-4-(2-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (500 mg, 1.2 mmol) in DMF (10 mL) to a solution of 2-(tert-butoxycarbonyl)amino)-4-(2-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (500 mg, 1.2 mmol) in DMF (10 mL). Stir the reaction mixture in a balloon at 0 °C to room temperature for 5 h under a nitrogen atmosphere. Monitor the reaction by LCMS. After completion, quench the reaction mixture with water (20 mL) and extract with EA (30 mL × 3). Combine the organic layers and wash with brine (20 mL × 3), then dry over anhydrous sodium sulfate. Remove the solvent under reduced pressure to give the product as a white solid (500 mg, 82% yield). LCMS: calculated exact mass = 496.1, found (ESI+) [M+H] + = 497.1.

[0582] Step 2: 2-Amino-4-(3-chlorophenyl)-N-(6-hydroxypyridin-3-yl)benzo[d]thiazol-6-carboxamide (Compound 35)

[0583]

[0584] A solution of 4M HCl in dioxane (10 mL) was added to a solution of (4-(3-chlorophenyl)-6-((6-hydroxypyridin-3-yl)carbamoyl)benzo[d]thiazo-2-yl)carbamate tert-butyl ester (500 mg, 1.0 mmol) in DCM (10 mL). The reaction mixture was stirred by a balloon at 0 °C to room temperature for 5 h under a N2 atmosphere. The reaction was monitored by LCMS. After completion, the solid was collected by filtration, washed with DCM (30 mL × 3) and dried under reduced pressure. The crude product was purified by MPLC (CH3OH / H2O) to give the product as a white solid (20 mg, 5% yield).

[0585] 1H NMR (400 MHz, DMSO) δ 11.53 (s, 1H), 9.99 (s, 1H), 8.26 (d, J =1.8 Hz, 1H), 7.98 (s, 2H), 7.95 (dd, J = 7.7, 2.3 Hz, 2H), 7.91 (t, J = 1.8Hz, 1H), 7.81 – 7.77 (m, 1H), 7.68 (dd, J = 9.7, 2.9 Hz, 1H), 7.51 (t, J =7.8 Hz, 1H), 7.44 (ddd, J = 8.0, 2.1, 1.1 Hz, 1H), 6.40 (d, J = 9.7 Hz, 1H).

[0586] Preparation of 4-(3-chlorophenyl)-N-(6-hydroxypyridin-3-yl)benzo[d]thiazol-6-carboxamide (compound 36)

[0587]

[0588] BTFFH (442 mg, 1.4 mmol), DIEA (225 mg, 1.7 mmol), and 5-aminopyridine-2(1H)-one (154 mg, 1.4 mmol) were added to a solution of 4-(3-chlorophenyl)benzo[d]thiazolyl-6-carboxylic acid (200 mg, 0.7 mmol) in DMF (10 mL). The reaction mixture was stirred by balloon at 0 °C to room temperature for 6 h under N2 atmosphere. The reaction was monitored by LCMS. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (20 mL × 3). The organic layers were combined, washed with brine (20 mL × 3), and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The reaction mixture was purified by MPLC (CH3OH / H2O) to give the product as a white solid (27 mg, 18% yield). LCMS: calculated exact mass = 381.0, found (ESI+) [M+H] + = 382.0.

[0589] 1H NMR (400 MHz, DMSO) δ 11.52 (s, 1H), 10.28 (s, 1H), 9.62 (s, 1H), 8.77 (s, 1H), 8.21 (s, 1H), 8.07 – 7.98 (m, 2H), 7.91 (d, J = 7.5 Hz, 1H), 7.71 (dd, J = 9.8, 2.5 Hz, 1H), 7.61 – 7.53 (m, 2H), 6.43 (d, J = 9.7 Hz, 1H).

[0590] Biological assay methods

[0591] The following embodiments are provided to illustrate this disclosure but do not limit it. Those skilled in the art will recognize that the following measurements and methods may be modified by selecting appropriate materials and methods.

[0592] Example 1

[0593] PDE4 enzyme assay-cAMP detection

[0594] The inhibition of cAMP production in human PDE4B was evaluated using Revvity's cAMP Gi kit as described by the manufacturer. The compound was serially diluted in 10-point increments in 100% DMSO, and 90 nL of each serially diluted compound was dispensed into 384-well assay plates (6007290, PerkinElmer) using a liquid handling instrument (mosquito® LV, sptlabtech). Using the Apricot S2 liquid handling instrument, 5 µL of the full-length PDE4 protein and the kinase domain of the PDE4 protein, diluted in reaction buffer (1X HBSS, 5 mM HEPES, 3 mM MgCl2, 0.1% BSA, pH 7.4), were transferred to each well to achieve final concentrations of 0.5 nM and 2.5 nM, respectively. The plates were then placed on a plate shaker and incubated at room temperature for 10 minutes. Dispense 5 μL of cAMP into each well to achieve a final concentration of 16 nM. After rotating the plate at 1000 rpm for 10 seconds, incubate at room temperature for 1 hour. Add a mixture of 5 μL of 1X cAMP Eu Cryptate working solution and 5 μL of 1X cAMP d2 antibody working solution to each well, diluted in lysis and detection buffer 8 and supplemented with 1 mM IBMX. Then, incubate the assay plate at room temperature for another 1 hour. Read the fluorescence using an EnVision plate reader (PerkinElmer), where excitation is at 330 nm and emission is at 615 nm and 665 nm.

[0595]

[0596] Table 3. Hill slope and IC50 for compounds targeting the full-length PDE4B2 protein and the PDE4B2 catalytic domain only. 50 value.

[0597]

[0598]

[0599] N / A = Unavailable

[0600] *Data provided by the second independent trial.

[0601] cAMP detection based on PDE4 cells

[0602] The inhibition of cAMP production by human PDE4 isoforms was evaluated using the Meso Scale Discovery cAMP kit as described by the manufacturer. HEK293 cells were seeded into 96-well plates and incubated at 37°C with 5% CO2 for 2 hours to ensure adhesion. PDE4B1 was transfected into HEK293 cells using Lipofectamine. The compounds were measured at a concentration of 10 µM in DMSO diluted with PBS. HEK293 control cells were used to determine the selectivity for PDE4A / PDE4D, due to the high endogenous levels of these isoforms as determined by qPCR. HEK293-PDE4B1 overexpression was used to determine the selectivity for PDE4B. Cells were pretreated with saliva extract for 1 hour, followed by treatment with a PDE4 inhibitor for 15 minutes, and then cAMP (µM) accumulation was measured using an MSD plate reader (Meso Scale Discovery). Results were presented in […]. Figure 1 As shown in the image.

[0603] Example 2

[0604] Animal models and their husbandry:

[0605] All experiments were conducted according to the institution's animal care guidelines. Scn1a+ / - and wild-type (WT) mice (B6129F1) were housed in temperature- and humidity-controlled rooms under a 12-hour light / dark cycle with free access to food and water. Kv1.1 (kcna1- / -) and GABA_A1 receptor (gabra1- / -) knockout zebrafish (Danio rerio) were similarly maintained. The zebrafish were housed in standard aquariums at 28°C under a 14-hour light / 10-hour dark cycle and fed twice daily.

[0606] Drug treatment:

[0607] Compound 1 in a 1:1:4:4 (DMSO: Tween 80: fractionated coconut oil: sterile water) medium was administered to mice twice daily via oral gavage at a dose of 40 mg / kg, or diluted to 50 μM in DMSO in E3 medium (5.0 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2, 0.33 mM MgSO4, and 0.1 mg / L methylene blue) for zebrafish studies. Positive controls included valproic acid (VPA, 250 mg / kg in mice and 50 μM in fish), carbamazepine (CBZ, 50 μM), levetiracetam (LEV, 50 μM), lamotrigine (LTG, 50 μM), and VPA + clonazan (VPA 50 μM + CLO, 50 μM). The medium groups received an equal volume of the drug solvent.

[0608] Monitoring of seizures in Scn1a+ / - mice via video EEG (vEEG) Figure 2A and Figure 2B ):

[0609] Scn1a + / - Mice were implanted with epidural EEG electrodes on day 18 after birth and continuously monitored for 10 days using a tethered video-EEG monitoring system (Pinnacle Technology, Inc.) starting on day 20 after birth. Drug treatment or a mediator was administered on the first day of vEEG monitoring and for the following 10 days. Video EEG data were acquired at a sampling rate of 250 Hz and bandpass filtered from 0.5 to 100 Hz. Scn1a was used in mice exposed to compound 1 or the mediator. + / - In mice, the frequency of quantified seizures (seizures per day) during a 10-day treatment period ( Figure 2A Seizures were scored using a modified Racine scale: 1. Oral and facial automatisms; 2. Head nodding; 3. Unilateral forelimb clonus; 4. Bilateral forelimb clonus with orthostatic disposition; 5. Generalized tonic-clonic seizures, orthostatic disposition, and falls; 6. Generalized tonic-clonic seizures with uncontrolled and erratic running and jumping. In separate analyses, mice exhibiting severe phenotypes (Racine stage ≥ 4 within the first 48 hours) were excluded from seizure activity and survival assays. Figure 2B ).

[0610] Motor epileptic seizure activity in hereditary and PTZ-induced zebrafish models ( Figure 3 ):

[0611] Zebrafish larvae with 5-day post-fertilization (dpf) kv1.1- / - (kcna1- / -) and GABA_A1- / - (gabra1- / -) knockout were acclimated for 30 minutes in a Zebrabox (ViewPoint Behavior Technology), and their seizure activity was assessed using recognized kinesiology methods. Larvae exhibiting confirmed seizure activity at baseline were used for further experiments. During the acclimation phase, 5-dpf tail-length (TL, wild-type) zebrafish larvae were exposed to PTZ (5 mM) in the Zebrabox for 30 minutes, and seizure activity was assessed. TL larvae with confirmed seizure activity at baseline were selected for the next stage of experiments. An additional group of TL larvae exposed to the medium were acclimated and used as a negative control for the experiments.

[0612] After acclimatization, baseline movement readings were taken from all zebrafish. Larvae were then exposed to compound 1, VPA, CBZ, LEV, LTG, or VPA + CLO for 1.5 hours, and test movement readings were collected. After the test readings, the larvae were transferred to fresh plates containing E3 for continued monitoring after drug elution.

[0613] Maximum electroconvulsive seizure (MES) model ( Figure 4 ):

[0614] WT mice approximately 60 days old were subjected to MES stimulation (60 Hz, 0.6 ms pulse width, 0.2 s duration, 50 mA) using ear clip electrodes. Compound 1 or the mediator was administered for 3 consecutive days, with the last dose given 40 minutes before the MES test. VPA was administered 10 minutes before the MES stimulation.

[0615] Survival analysis ( Figure 5A and Figure 5B ):

[0616] Survival of Scn1a+ / - mice was monitored in their cages after removal of the vEEG probe. Kaplan-Meier survival analysis was performed to compare the compound 1 group with the mediator group. Median survival after treatment cessation was also assessed.

[0617] Barnes Maze Test for Learning, Memory, and Search Strategies ( Figure 6A ):

[0618] Over a 4-day period, an accelerated Barnes maze protocol was used to train approximately 60-day-old WT mice exposed to either the medium or compound 1, and Scn1a+ / - mice exposed to either the medium or compound 1, with a probe test performed on day 5. On day 1, mice were acclimatized to the maze and escape box. From days 2 to 4, mice underwent four 3-minute training trials with 15-minute intervals, for a total of 12 training trials. Learning curves were generated and denoted as "escape latency" during each trial. On day 5 (24-hour probe test), the escape box was removed, and mice were given 1 minute of free exploration of the maze. The time spent near the "target hole" during the probe test was quantified after treatment with compound 1 or the medium.

[0619] Y-maze test for spatial recognition and memory ( Figure 6B ):

[0620] A forced Y-maze was used to test spatial recognition memory in mice. In the training test, mice were placed in a Y-maze with one arm blocked and allowed free exploration for 5 minutes before being returned to their enclosure. After a 30-minute test interval, the blocked arm of the Y-maze was opened, and the mice were returned for a 5-minute probe test. The number of new arms entered was recorded as an indicator of working memory and spatial recognition in WT and Scn1a+ / - mice treated with Compound 1 or the mediator.

[0621] Marble Burial Test for Anxiety and Repetitive Behaviors ( Figure 7A ):

[0622] A marble burial test was performed to evaluate repetitive and anxiety-related behaviors in WT and Scn1a+ / - epileptic mice treated with compound 1 or the mediator. Mice were placed in static cages with 18 evenly spaced glass marbles arranged in 5 cm of clean bedding and allowed to explore for 30 minutes. The number of buried marbles (≥2 / 3 covered) was quantified. A large number of buried marbles was an indicator of repetitive and anxiety-like behaviors.

[0623] Open field test using new objects ( Figure 7B and Figure 7C ):

[0624] The open field test with a novel object was used to evaluate the motor activity, exploratory, and anxiety-like behaviors of WT mice exposed to Compound 1 or the medium. Mice were placed in a moderately lit 50 × 50 cm arena where the novel object was positioned in the center. Behavior was recorded over 10 minutes, measuring total distance traveled and time spent in the central area. Increased time spent in the center was interpreted as reduced anxiety, while reduced movement indicated a potential sedative effect or altered exploratory behavior.

[0625] Statistical analysis:

[0626] Data were analyzed using GraphPad Prism. Statistical significance was assessed, where applicable, using two-way ANOVA with Sidak post-hoc test, Kaplan-Meier survival analysis, and unpaired t-tests. Data are presented as mean ± SEM, where p-values ​​are indicated (p < 0.05 was considered significant).

[0627] By incorporating references

[0628] All publications and patents mentioned herein are incorporated herein by reference in their entirety, as if each individual publication or patent were specifically and individually indicated to be incorporated herein by reference. In case of conflict, the provisions of this application, including any definitions herein, shall prevail.

[0629] equivalent

[0630] While specific embodiments of the subject matter have been discussed, the foregoing description is illustrative and not restrictive. Many variations of this disclosure will become apparent to those skilled in the art upon reading this specification and the following claims. The full scope of this disclosure should be determined by reference to the full scope of the claims and their equivalents, as well as the description and such variations.

Claims

1. A compound of formula (I), （I） Or pharmaceutically acceptable salts, wherein: R 1 It is an amino, H, or alkyl group; R 2 It is a heteroaryl, heterocyclic, cycloalkyl, or aryl group; and R 3 It is aryl, heterocyclic, cycloalkyl, or heteroaryl.

2. The compound according to claim 1, wherein R 2 It is a heteroaryl group.

3. The compound according to claim 1 or 2, wherein R 2 It is pyridyl, pyridone, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, or oxazolyl.

4. The compound according to any one of claims 1-3, wherein R 2 It is pyridyl (e.g., 3-pyridyl).

5. The compound according to any one of claims 1-4, wherein R 2 Optionally substituted with amino (e.g., -NH2, morpholino, or piperazine) or alkyl (e.g., C1-C6 alkyl).

6. The compound according to any one of claims 1-5, wherein R 2 Choose from the following groups: , , , , , , , , , and .

7. The compound according to any one of claims 1-6, wherein R 3 It may optionally be substituted with halogen, cyano, sulfone, nitro, ketone or alkyl.

8. The compound according to any one of claims 1-7, wherein R 3 It is an aryl group.

9. The compound according to any one of claims 1-8, wherein R 3 It is a phenyl group, wherein each hydrogen atom in the phenyl group is independently and optionally substituted with a halogen, cyano, sulfone, nitro, ketone or alkyl group.

10. The compound according to any one of claims 1-9, wherein R 3 It is a phenyl group, wherein each hydrogen atom in the phenyl group is independently and optionally replaced by a halogen (e.g., chlorine).

11. The compound according to any one of claims 1-10, wherein R 3 It is 3-chlorophenyl.

12. The compound according to any one of claims 1-9, wherein R 3 It is a phenyl group, wherein each hydrogen atom in the phenyl group is independently and optionally substituted with a nitro group.

13. The compound according to any one of claims 1-9 or 12, wherein R 3 It is 3-nitrophenyl.

14. The compound according to any one of claims 1-7, wherein R 3 It is a heteroaryl group.

15. The compound according to any one of claims 1-7 or 14, wherein R 3 It is a thiophene group, wherein each hydrogen atom in the thiophene group is independently and optionally replaced by a halogen (e.g., chlorine).

16. The compound according to any one of claims 1-7, wherein R 3 It is a heterocyclic group.

17. The compound according to any one of claims 1-7 or 16, wherein R 3 It is tetrahydropyranyl, dihydropyranyl, piperidinyl, or tetrahydropyridinyl.

18. The compound according to any one of claims 1-7 or 16-17, wherein R 3 It is piperidinyl or tetrahydropyridinyl, wherein each hydrogen atom in the piperidinyl or tetrahydropyridinyl group is independently and optionally substituted with an alkyl group (e.g., C1-C6 alkyl group).

19. The compound according to any one of claims 1-7, wherein R 3 It is a cycloalkyl group.

20. The compound according to any one of claims 1-7 or 19, wherein R 3 It is a cyclohexyl group, wherein each hydrogen atom in the cyclohexyl group is independently and optionally replaced by a halogen (e.g., fluorine).

21. The compound according to any one of claims 1-7, wherein R 3 Choose from the following groups: , , , , , , , , , , and .

22. The compound according to any one of the preceding claims, wherein R 1 It is H or NH2.

23. The compound according to claim 1, wherein the compound has formula (II): (II), Or its pharmaceutically acceptable salt, wherein: R 2 It is a heteroaryl or aryl; and R 3 It is a heterocyclic group, cycloalkyl group, heteroaryl group, or aryl group.

24. The compound according to claim 1, wherein the compound has formula (III): (III), Or its pharmaceutically acceptable salt, wherein: X 1 and X 3 Each of them is independently CH or N; X 2 It is CH or C-NH2; R 1 It is H, amino, or alkyl; and R 3 It is a heterocyclic group, cycloalkyl group, heteroaryl group, or aryl group; The condition is X 1 and X 3 At least one of them is not N.

25. The compound according to claim 1, wherein the compound has formula (IIIa): (IIIa), Or its pharmaceutically acceptable salt, wherein: X 1 and X 3 Each of them is independently CH or N; R 1 It is H, amino, or alkyl; R 3 It is a heterocyclic group, cycloalkyl group, heteroaryl group, or aryl group; and R 5 It is H or amino; The condition is X 1 and X 3 At least one of them is not N.

26. The compound according to claim 1, wherein the compound has the formula (IIIb): (IIIb), Or its pharmaceutically acceptable salt, wherein: R 1 It is H, amino, or alkyl; R 4 It is a halogen, cyano, sulfone, nitro, ketone, or alkyl group; and R 5 It is H or amino.

27. The compound according to claim 1, wherein the compound has formula (IV): (IV) Or its pharmaceutically acceptable salt, wherein: R 4 It is a halogen, cyano, sulfone, nitro, ketone or alkyl group.

28. The compound according to claim 1, selected from the group consisting of: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and , Or its pharmaceutically acceptable salt.

29. The compound according to claim 1, selected from the group consisting of: , , , , , , , and , Or its pharmaceutically acceptable salt.

30. The compound according to any one of the preceding claims, wherein the compound is a pharmaceutically acceptable salt.

31. A pharmaceutical composition comprising a compound according to any one of claims 1-30 and a pharmaceutically acceptable excipient.

32. A method of treating a patient’s disease, the method comprising administering to the patient in need an effective amount of the compound according to any one of claims 1-30.

33. The method of claim 32, wherein the disease is an inflammatory disease selected from the group consisting of pulmonary inflammatory diseases, dermatological inflammatory diseases, and neurological inflammatory diseases.

34. The method of claim 33, wherein the disease is an inflammatory disease selected from the group consisting of asthma, chronic obstructive pulmonary disease (COPD), psoriasis, atopic dermatitis, inflammatory bowel disease, and rheumatoid arthritis.

35. The method of claim 32, wherein the disease is a cognitive or emotional disorder selected from the group consisting of learning disabilities, memory loss, and Fragile X syndrome.

36. The method of claim 32, wherein the disease is cancer.

37. The method of claim 36, wherein the cancer is a solid tumor.

38. The method of claim 36, wherein the cancer is esophageal cancer, brain cancer, pancreatic cancer, colon cancer, hematologic cancer, lung cancer, prostate cancer, skin cancer, head and neck cancer, CNS cancer, gastric cancer, breast cancer, bladder cancer, or ovarian cancer.

39. The method of claim 32, wherein the disease is a central nervous system disorder (e.g., epilepsy).

40. The method of claim 39, wherein the central nervous system disorder is a neurodevelopmental disorder (e.g., epilepsy, autism, attention deficit disorder (ADD) or attention deficit hyperactivity disorder (ADHD)), an acute neurological disorder (e.g., stroke or traumatic brain injury), a neurodegenerative disorder (e.g., Alzheimer's disease, dementia, multiple sclerosis or Huntington's disease), or a psychological / mental disorder (e.g., schizophrenia, depression, anxiety disorder (e.g., generalized anxiety disorder or social anxiety disorder), bipolar disorder, addiction or obsessive-compulsive disorder (OCD)).

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