阅读说明：本技术 取代的-3H-咪唑并[4,5-c]吡啶和1H-吡咯并[2,3-c]吡啶系列的新型外核苷酸焦磷酸酶/磷酸二酯酶-1(ENPP1)和干扰素基因刺激物(STING)调节剂作为癌症免疫治疗剂 (Novel ectonucleotide pyrophosphatase/phosphodiesterase-1 (ENPP1) and interferon gene Stimulator (STING) modulators of substituted-3H-imidazo [4,5-c ] pyridines and 1H-pyrrolo [2,3-c ] pyridines series) 是由 哈里普拉萨德·梵卡亚拉帕提 刘晓慧 古鲁桑卡·拉玛穆尔蒂 苏尼尔·沙玛 莫汉·拉奥·卡迪奇 于 2018-07-27 设计创作，主要内容包括：取代的-3H-咪唑并[4,5-c]吡啶和1H-吡咯并[2,3-c]吡啶系列的新型外核苷酸焦磷酸酶/磷酸二酯酶-1(ENPP1)和相关化合物,它们可用作ENPP1的抑制剂；制备这些化合物的合成方法；包含所述化合物的药物组合物；以及使用所述化合物和组合物治疗与ENPP1功能障碍相关的病症的方法。(Novel exonucleotide pyrophosphatase/phosphodiesterase-1 (ENPP1) and related compounds of the substituted-3H-imidazo [4,5-c ] pyridine and 1H-pyrrolo [2,3-c ] pyridine series, useful as inhibitors of ENPP 1; synthetic methods for preparing these compounds; pharmaceutical compositions comprising said compounds; and methods of using the compounds and compositions to treat conditions associated with ENPP1 dysfunction.)

1. A compound of formula I:

wherein

Y is S;

z is-H or-N;

X₁is-C ═ O or-NH,

X₂is-C ═ O or-NH, with the proviso that if X₁is-C ═ O, then X₂is-NH; and if X₁is-NH, then X₂is-C ═ O;

R₁selected from the group consisting of: -H, halogen, -OCF₃、-CF₃、-CN、OCH₃and-CH₃；

R₂Selected from the group consisting of: -H, -NH₂-OH and-CH₃；

R₃Selected from the group consisting of: -H, -NH₂-OH and-CH₃；

R₄Selected from the group consisting of: -H, -CH₂-CH₃and-CH₃；

R₅Selected from the group consisting of:

R₆Selected from the group consisting of: -OCH₃、OH、F、Cl、-CH₃、O-CH₂-CH₃And OCH₂-CF₃、OCHF₂、

Or an isomer or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein

Z is-N;

X₁is a compound of formula-C ═ O,

X₂is-NH;

R₁selected from the group consisting of: halogen and-CN;

R₂selected from the group consisting of: -H and-CH₃；

R₃is-H;

R₄is-H;

R₅is that

R₆Selected from the group consisting of: -OCH₃And an OH group, and a nitrogen-containing compound,

or an isomer or a pharmaceutically acceptable salt thereof.

3. The compound of claim 2, wherein R₁Selected from the group consisting of: cl and-CN.

4. According to claimThe compound of claim 1, wherein the compound is selected from the group consisting of:

5. a pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 and a pharmaceutically acceptable carrier.

6. A method of treating an uncontrolled cell proliferation disorder in a mammal, the method comprising the step of administering to the mammal an effective amount of a compound of claim 1.

7. A method of treating cancer in a mammal comprising the step of administering to said mammal an effective amount of a compound of claim 1.

Background

Members of the ectonucleotide pyrophosphatase/phosphodiesterase (ENPP) family include 7 subtypes, ENPP1-7, which are type II transmembrane glycoproteins or extracellular enzymes. The results of mass spectrometry and proteomics analyses from 370 multiple protein targets led to the identification of the extracellular protein ENPP1 as one of the hot proteins exhibiting high hydrolytic activity. ATP is an identified substrate of ENPP1, which is hydrolyzed to AMP and PPi. CD73 converts AMP into adenosine and inorganic Phosphate (PI). Kinetic experimental data indicate that ENPP1 is capable of hydrolyzing ATP. These exonucleotidases are involved in the hydrolysis of pyrophosphate (PPi) and phosphodiester bonds in extracellular nucleotides (e.g., triphosphates, oligonucleotides) and produce 5' -monophosphate nucleosides. One of the key subtypes, ENPP1 (plasma cell membrane glycoprotein-1, PC-1), is involved in many physiological processes, such as development, formation and transport, as well as pathophysiological conditions. Abnormal ENPP1 expression was detected in breast cancer relative to normal breast epithelium, which is evidence of its potential in the development of bone metastases (occurring in about 80% of cases), hodgkin lymphoma, hepatocellular carcinoma, follicular lymphoma, glioblastoma, and other malignant tumor tissues.

Recent reports have shown that Cyclic Dinucleotides (CDNs), the substrate of ENPP1, stimulate innate immunity through STING-dependent interferon gene activation. Similar to checkpoint inhibitors such as anti-PD-1 or PD-L1, which are promising immunotherapeutic agents for various cancers, inhibition of STING pathway activation by ENPP1 is crucial for tumor control. Furthermore, mutations in ENPP1 have been associated with a variety of conditions, including infant artery calcification (infant generalized artery calcification or GACI), posterior longitudinal ligament ossification of the spine, and insulin signaling and resistance. ENPP1 is highly expressed in bone and cartilage and is associated with pulmonary and renal fibrosis. The expression of ENPP1 also correlated with the classification of astrocytic tumors. Another study reported that ENPP1 was required to maintain the undifferentiated and proliferative state of glioblastoma stem-like cells. Thus, ENPP1 is an attractive drug target for the development of novel anti-cancer, cardiovascular, diabetes, obesity and anti-fibrotic therapies.

The importance of ENPP1 activity was further investigated both in direct binding assays and in vitro cell efficacy on MDA-MB231 cells. In cell-specific and in vivo experiments, the ENPP1 knockdown based on siRNA significantly reduced its catalytic activity. These experiments demonstrated that ENPP1 activity was abolished upon treatment with siRNA. This further supports the effectiveness of the target in certain diseases. It has recently been shown that bisphosphonate analogues of endogenous cGAMP are resistant to hydrolysis by ENPP1 phosphodiesterase, in particular Cyclic Dinucleotides (CDNs) are more effective in inducing IFN- β secretion in human THP1 cells by a mechanism that inhibits ENPP1 activity and simultaneous STING activation.

There is substantial evidence that ENPP1 is significantly expressed in human primary breast tumors, with the highest levels observed in breast cancer bone metastases relative to normal breast epithelium. These data support not only the potential role of ENPP1 in bone metastasis of breast cancer, but also its potential prognostic marker for breast cancer. These results from the target validation experiments clearly support the pharmacological role of ENPP1 in the development of new cancer immunotherapies.

Furthermore, ENPP1 activity is also associated with bacterial and/or viral-caused diseases, and therefore modulators of ENPP1 are useful in the treatment of bacterial and/or viral diseases and conditions.

Disclosure of Invention

In one aspect, the invention relates to compounds of formula I:

wherein

Y is S;

z is-H or-N;

X₁is-C ═ O or-NH,

X₂is-C ═ O or-NH, with the proviso that if X₁is-C ═ O, then X₂is-NH; and if X₁is-NH, then X₂is-C ═ O;

R₁selected from the group consisting of: -H, halogen, -OCF₃、-CF₃、-CN、OCH₃and-CH₃；

R₂Selected from the group consisting of: -H, -NH₂-OH and-CH₃；

R₃Selected from the group consisting of: -H, -NH₂-OH and-CH₃；

R₄Selected from the group consisting of: -H, -CH₂-CH₃and-CH₃；

R₅Selected from the group consisting of:

and R is₆Selected from the group consisting of: -OCH₃、OH、F、Cl、-CH₃、O-CH₂-CH₃And OCH₂-CF₃、OCHF₂Or an isomer or a pharmaceutically acceptable salt thereof.

In a preferred embodiment of the process according to the invention,

z is-N;

X₁is a compound of formula-C ═ O,

X₂is-NH;

R₁selected from the group consisting of: halogen and-CN;

R₂selected from the group consisting of: -H and-CH₃；

R₃is-H;

R₄is-H;

R₅is thatAnd is

R₆Selected from the group consisting of: -OCH₃And OH;

in another preferred embodiment, R₁Selected from the group consisting of: -Cl and-CN.

Examples of compounds provided include:

the invention also includes hydrates, solvates, polymorphs, isomers, tautomers, pharmaceutically acceptable salts of said compounds and pharmaceutically acceptable salts of tautomers of said compounds.

The invention also provides pharmaceutical formulations, medicaments comprising said compounds; methods of preparing pharmaceutical formulations, medicaments, compounds; 0 and methods of treating patients using the provided pharmaceutical formulations and compounds.

The compounds of the invention are identified by structure-based calculation of docking and binding free energies.

Also disclosed are pharmaceutical compositions comprising a therapeutically effective amount of the disclosed compounds and a pharmaceutically acceptable carrier.

Synthetic methods for preparing the disclosed compounds are also disclosed. In a further aspect, the products of the disclosed synthetic methods are disclosed.

Also disclosed are methods of treating a disorder associated with dysfunction of ENPP1 activity in a mammal, comprising the step of administering to the mammal a therapeutically effective amount of a disclosed compound, or a pharmaceutically acceptable salt, tautomer, isomer, hydrate, solvate, or polymorph thereof.

Also disclosed are methods of inhibiting ENPP1 activity in a mammal, comprising the step of administering to the mammal a therapeutically effective amount of at least one disclosed compound, or a pharmaceutically acceptable salt, tautomer, isomer, hydrate, solvate, or polymorph thereof.

Also disclosed are methods of inhibiting ENPP1 activity in at least one cell, the method comprising the step of contacting the at least one cell with an effective amount of at least one disclosed compound, or a pharmaceutically acceptable salt, tautomer, isomer, hydrate, solvate, or polymorph thereof.

Also disclosed are methods of treating a disorder associated with dysfunction of ENPP1 activity in a mammal by eliciting an immunotherapeutic response in the mammal, comprising administering to the mammal a therapeutically effective amount of a disclosed compound, or a pharmaceutically acceptable salt, tautomer, isomer, hydrate, solvate, or polymorph thereof, wherein the compound elicits an immunotherapeutic response beneficial for treating a disorder associated with ENPP1 activity. Such a condition may be, but is not limited to, any type of cancer or any disease caused by bacteria and/or viruses in which ENPP1 activity is implicated.

Also disclosed are pharmaceutical compositions comprising a pharmaceutically acceptable carrier and an effective amount of the disclosed compounds, or pharmaceutically acceptable salts, tautomers, isomers, hydrates, solvates, or polymorphs thereof.

Also disclosed are kits comprising at least one of the disclosed compounds or a pharmaceutically acceptable salt, tautomer, isomer, hydrate, solvate, or polymorph thereof.

Also disclosed are methods for manufacturing a medicament, comprising combining at least one disclosed compound or at least one disclosed product with a pharmaceutically acceptable carrier or diluent. In a further aspect, the invention relates to the use of a disclosed compound in the manufacture of a medicament for the treatment of a condition associated with dysfunction of ENPP1 activity. In a further aspect, the invention relates to the use of a disclosed compound in the manufacture of a medicament for the treatment of an uncontrolled cell proliferation disorder.

Also disclosed is the use of the disclosed compounds or the disclosed products in the manufacture of a medicament for treating a condition associated with ENPP1 dysfunction in a mammal.

Although aspects of the present invention may be described and claimed in a particular legal class, such as system legal class, this is for convenience only and those skilled in the art will understand that each aspect of the present invention may be described and claimed in any legal class. Unless expressly stated otherwise, it is in no way intended that any method or aspect set forth herein be required that its steps be performed in a specific order. Accordingly, where method claims do not specifically state in the claims or descriptions that the steps are to be limited to a particular order, it is not intended that an order be inferred, in any way. This applies to any possible non-express basis for interpretation, including logical problems associated with the arrangement of steps or operational flows, ordinary meaning as derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

Drawings

Figure 1 is a graph of ENPP1 inhibition assay of TMP of some compounds of the invention.

FIG. 2A is a graph of a positive control in the ENPP2 inhibition assay.

Figure 2B is a graph of ENPP2 inhibition assays for some compounds of the present invention.

FIG. 3 is a graph of ENPP1 Thermal transition analysis (Thermal Shift Assay) of the compounds of the present invention.

FIG. 4 is a bar graph of ENPP1 mineralization analysis of some compounds of the invention.

Detailed Description

A. Definition of

As used herein, the nomenclature of compounds (including organic compounds) can be given using the nomenclature suggested by the common name, IUPAC, IUBMB, or CAS. When one or more stereochemical features are present, the Cahn-ingold-Prelog rule of stereochemistry may be employed to specify stereochemical priorities, E/Z specifications, and the like. Given a name, one skilled in the art can perform systematic reduction of compound structure by using naming rules, or by commercially available software (e.g., ChemDraw)^TM(Cambridge Soft Corporation, U.S.A.)) to easily determine the structure of the compound.

As used in the specification and the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a functional group," "an alkyl group," or "a residue" includes mixtures of two or more such functional groups, alkyl groups, or residues, and the like.

Ranges can be expressed herein as from "about" one particular value, and/or to "about" another particular value. When such a range is expressed, another aspect includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another aspect. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It will also be understood that a number of values are disclosed herein, and that each value is also disclosed herein as "about" that particular value, in addition to the value itself. For example, if the value "10" is disclosed, then "about 10" is also disclosed. It is also understood that each unit between two particular units is also disclosed. For example, if 10 and 15 are disclosed, 11, 12, 13 and 14 are also disclosed.

References in the specification and concluding claims to parts by weight of a particular element or component in a composition, denotes the weight relationship between the element or component and any other elements or components in the composition or article for which parts by weight are indicated. Thus, in a compound containing 2 parts by weight of component X and 5 parts by weight of component Y, X and Y are present in a ratio of 2: 5, and is present in such a ratio regardless of whether additional components are present in the compound.

Unless expressly stated to the contrary, the weight percent (wt.%) of a component is based on the total weight of the formulation or composition in which the component is included.

As used herein, the term "ENPP 1" refers to ectonucleotide pyrophosphatase/phosphodiesterase.

As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

As used herein, the term "subject" may be a vertebrate, such as a mammal, fish, bird, reptile, or amphibian. Thus, the subject of the methods disclosed herein can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig, or rodent. The term does not denote a particular age or gender. Thus, adult and neonatal subjects, as well as fetuses (whether male or female) are intended to be covered. In one aspect, the subject is a mammal. "patient" refers to a subject having a disease or disorder. The term "patient" includes human and veterinary subjects. In some aspects of the disclosed methods, prior to the administering step, the subject has been diagnosed as in need of treatment for a disorder of uncontrolled cellular proliferation associated with dysfunction of ENPP 1. In some aspects of the disclosed methods, prior to the administering step, the subject has been diagnosed as in need of inhibition of ENPP 1.

As used herein, the term "treatment" refers to the medical management of a patient with the purpose of curing, ameliorating, stabilizing or preventing a disease, pathological condition or disorder. The term includes active treatment, i.e. treatment specifically directed to the improvement of a disease, pathological condition or disorder, and also includes causal treatment, i.e. treatment directed to the elimination of the cause of the associated disease, pathological condition or disorder. Furthermore, the term includes palliative treatment, i.e., treatment intended to alleviate symptoms rather than cure a disease, pathological condition, or disorder; prophylactic treatment, i.e. treatment aimed at minimizing or partially or completely inhibiting the development of the associated disease, pathological condition or disorder; and supportive therapy, i.e., therapy for supplementing another specific therapy for improvement of the associated disease, pathological condition or disorder. In various aspects, the term encompasses any treatment of a subject, including a mammal (e.g., a human), and includes (i) preventing the disease from occurring in a subject that may be predisposed to the disease but has not yet been diagnosed as having the disease; (ii) inhibiting, i.e. arresting the development of, said disease; or (iii) ameliorating the disease, i.e., causing regression of the disease. In one aspect, the subject is a mammal, e.g., a primate, and in a further aspect, the subject is a human. The term "subject" also includes domestic animals (e.g., cats, dogs, etc.), livestock (e.g., cows, horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mice, rabbits, rats, guinea pigs, drosophila, zebrafish, etc.).

As used herein, the terms "prevent" or "preventing" mean to exclude, avoid, eliminate, prevent, stop, or hinder something from happening, particularly by acting in advance. It is understood that where "reduce," "inhibit," or "prevent" is used herein, the use of two other words is also expressly disclosed unless expressly stated otherwise.

As used herein, the term "diagnosed" refers to having been physically examined by a skilled artisan (e.g., a physician) and found to have a condition that can be diagnosed or treated by a compound, composition, or method disclosed herein. For example, "diagnosed as having an uncontrolled cell proliferation disorder" refers to a disorder that has been physically examined by a skilled artisan (e.g., a physician) and found to be diagnosed or treated by a compound or composition that inhibits ENPP 1. As another example, "diagnosed as requiring inhibition of ENPP 1" refers to having been physically examined by a technician (e.g., a physician) and found to have a condition characterized by ENPP1 dysfunction. As discussed herein, such diagnosis may involve disorders, such as those of uncontrolled cellular proliferation, cancer, and the like. For example, "diagnosed as in need of treatment for one or more uncontrolled cell proliferation disorders associated with ENPP1 dysfunction" refers to a disorder that has been physically examined by a skilled person (e.g., a physician) and found to have one or more uncontrolled cell proliferation associated with ENPP1 dysfunction.

As used herein, the phrase "identified as in need of treatment for a condition" and the like refers to the selection of a subject based on the need to treat a disease. For example, based on an early diagnosis by a skilled artisan, a subject can be identified as having a need to treat a disorder (e.g., a disorder associated with dysfunction of ENPP1), and then treat the disorder. In one aspect, it is contemplated that the identifying can be performed by a person other than the person making the diagnosis. In a further aspect, it is also contemplated that the administration can be by a person who subsequently performs the administration.

As used herein, the terms "administering" and "administration" can refer to any method of providing a pharmaceutical formulation to a subject. Such methods are well known to those skilled in the art and include, but are not limited to, oral administration, transdermal administration, inhalation administration, nasal administration, topical administration, intravaginal administration, ocular administration, intra-aural administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, intraurethral administration, and parenteral administration, including injectable, such as intravenous administration, intraarterial administration, intramuscular administration, and subcutaneous administration. Administration may be continuous or intermittent. In various aspects, the formulation can be administered therapeutically; i.e., for treating an existing disease or disorder. In further various aspects, the formulation may be administered prophylactically; i.e., for the prevention of a disease or disorder.

The term "contacting" as used herein refers to bringing the disclosed compounds and cells, target receptors, or other biological entities together such that the compounds are capable of affecting the activity of a target (e.g., receptor, cell, etc.) either directly (i.e., by interacting with the target itself), or indirectly (i.e., by interacting with another molecule, cofactor, factor, or protein upon which the activity of the target depends).

As used herein, the terms "effective amount" and "dose effective" refer to an amount sufficient to achieve a desired result or effect on an undesired condition. For example, a "therapeutically effective amount" refers to an amount sufficient to achieve a desired therapeutic result or effect on an undesirable condition, but generally insufficient to cause an adverse side effect. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the condition being treated and the severity of the condition; the specific ingredients used; the age, weight, general health, sex, and diet of the patient; the time of administration; the route of administration; the rate of excretion of the particular compound used; the duration of the treatment; drugs used in combination or concomitantly with the specific compound employed and similar factors well known in the medical arts. For example, one skilled in the art will start doses of the compound at levels below those required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. If desired, the effective daily dose may be divided into multiple doses for administration purposes. Thus, a single dose composition may contain such amounts or submultiples thereof to make up the daily dose. In the case of contraindications, the dosage may be adjusted by the individual physician. The dosage may vary, and may be administered once or multiple times daily for one or more days. Guidelines for appropriate dosing of a given class of pharmaceutical products can be found in the literature. In further various aspects, the formulation can be administered in a "prophylactically effective amount"; i.e., an amount effective to prevent a disease or disorder.

"EC" as used herein₅₀By "is meant a concentration of a substance (e.g., a compound or drug) that is 50% required to agonize or activate a biological process or process component (including proteins, subunits, organelles, ribonucleoproteins, etc.). In one aspect, as further defined elsewhere herein, EC₅₀And may refer to the concentration of a substance required for 50% agonism or activation in the body. In a further aspect, EC₅₀Refers to a concentration of agonist or activator that elicits a response halfway between baseline and maximal responseAnd (4) degree.

As used herein, "IC₅₀By "is meant the concentration of a substance (e.g., a compound or drug) required to inhibit 50% of a biological process or process component (including proteins, subunits, organelles, ribonucleoproteins, etc.). For example, as further defined elsewhere herein, an IC₅₀It may refer to the concentration of substance required for 50% inhibition in vivo or for inhibition measured in vitro. Or, IC₅₀Refers to the half maximal (50%) Inhibitory Concentration (IC) of the substance. Inhibition can be measured in cell lines such as AN3 CA, BT-20, BT-549, HCT 116, HER218, MCF7, MDA-MB-231, MDA-MB-235, MDA-MB-435S, MDA-MB-468, PANC-1, PC-3, SK-N-MC, T-47D, and U-87 MG.

The term "pharmaceutically acceptable" describes materials that are not biologically or otherwise undesirable, i.e., do not cause unacceptable adverse biological effects or interact in a deleterious manner.

The term "stable" as used herein refers to a compound that is substantially unchanged when subjected to conditions that allow its production, detection, and in some aspects its recovery, purification, and use for one or more of the purposes disclosed herein.

As used herein, the term "derivative" refers to a compound having a structure derived from a parent compound (e.g., a compound disclosed herein) and which is sufficiently similar in structure to the compound disclosed herein that, based on the similarity, one of skill in the art would expect it to exhibit the same or similar activity and utility as, or to induce as a precursor the same or similar activity and utility as, the claimed compound. Exemplary derivatives include salts, esters, amides, salts of esters or amides, and N-oxides of the parent compound.

The term "pharmaceutically acceptable carrier" as used herein refers to sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. These compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. Prevention of the action of microorganisms can be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin. Injectable depot forms are prepared by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, poly (orthoesters), and poly (anhydrides). Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues. The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. Suitable inert carriers may include sugars such as lactose. Desirably, at least 95% by weight of the active ingredient particles have an effective particle size in the range of 0.01 to 10 microns.

As used in the specification and concluding claims, a residue of a chemical refers to the moiety of the resulting product that is a chemical in a particular reaction scheme or subsequent formulation or chemical product, regardless of whether the moiety is actually obtained from the chemical. Thus, an ethylene glycol residue in a polyester refers to one or more-OCH's in the polyester₂CH₂O-units, regardless of whether ethylene glycol is used to make the polyester. Similarly, a sebacic acid residue in a polyester refers to one or more-CO (CH) residues in the polyester₂)₈CO-moiety, and whether or not the residue isIs obtained by reacting sebacic acid or an ester thereof to obtain a polyester.

As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. For suitable organic compounds, the permissible substituents can be one or more and the same or different. For purposes of this disclosure, a heteroatom (e.g., nitrogen) may have a hydrogen substituent and/or any permissible substituents of organic compounds described herein that satisfy the valencies of the heteroatom. The present disclosure is not intended to be limited in any way by the permissible substituents of organic compounds. Likewise, the terms "substituted" or "substituted with … …" include the implicit proviso that such substitution is in accordance with the permitted valences of the substituted atoms and substituents, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, and the like. It is also contemplated that, in certain aspects, individual substituents may be further optionally substituted (i.e., further substituted or unsubstituted), unless explicitly stated to the contrary.

In defining the terms, "A" and "A" are used¹”、“A²”、“A³"and" A⁴"is used herein as a generic symbol to denote various specific substituents. These symbols may be any substituent, not limited to those disclosed herein, and when they are defined as certain substituents in one instance, they may be defined as others in another instance.

The term "alkyl" as used herein is a branched or unbranched saturated hydrocarbon group having 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like. The alkyl group may be cyclic or acyclic. The alkyl group may be branched or unbranched. Alkyl groups may also be substituted or unsubstituted. For example, an alkyl group may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol as described herein. A "lower alkyl" group is an alkyl group containing 1-6 (e.g., 1-4) carbon atoms.

For example, a "C1-C3 alkyl" group can be selected from methyl, ethyl, n-propyl, isopropyl, and cyclopropyl, or from a subset thereof. In certain aspects, a "C1-C3 alkyl" group can be optionally further substituted. As another example, a "C1-C4 alkyl" group can be selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and cyclobutyl, or from a subset thereof. In certain aspects, a "C1-C4 alkyl" group can be optionally further substituted. By way of further example, a "C1-C6 alkyl" group can be selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, sec-pentyl, tert-pentyl, neopentyl, cyclopentyl, n-hexyl, isohexyl, 3-methylpentane, 2, 3-dimethylbutane, neohexane, and cyclohexane, or a subset thereof. In certain aspects, a "C1-C6 alkyl" group can be optionally further substituted. As another example, a "C1-C8 alkyl" group can be selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, sec-pentyl, tert-pentyl, neopentyl, cyclopentyl, n-hexyl, isohexyl, 3-methylpentane, 2, 3-dimethylbutane, neohexane, cyclohexane, heptane, cycloheptane, octane, and cyclooctane, or a subset thereof. In certain aspects, a "C1-C8 alkyl" group can be optionally further substituted. As another example, a "C1-C12 alkyl" group can be selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, sec-pentyl, tert-pentyl, neopentyl, cyclopentyl, n-hexyl, isohexyl, 3-methylpentane, 2, 3-dimethylbutane, neohexane, cyclohexane, heptane, cycloheptane, octane, cyclooctane, nonane, cyclononane, decane, cyclodecane, undecane, cycloundecane, dodecane, and cyclododecane, or a subset thereof. In certain aspects, a "C1-C12 alkyl" group can be optionally further substituted.

Throughout the specification, "alkyl" is generally used to refer to both unsubstituted alkyl and substituted alkyl; however, substituted alkyl groups are also specifically mentioned herein by identifying specific substituents on the alkyl group. For example, the term "halogenated alkyl" or "haloalkyl" particularly refers to an alkyl group substituted with one or more halides, such as fluorine, chlorine, bromine, or iodine. As described below, the term "alkoxyalkyl" specifically refers to an alkyl group substituted with one or more alkoxy groups. As described below, the term "alkylamino" specifically refers to an alkyl group substituted with one or more amino groups, and the like. When "alkyl" is used in one instance and a specific term such as "alkyl alcohol" is used in another instance, this does not mean that the term "alkyl" does not refer to the specific term such as "alkyl alcohol" or the like.

This approach is also applicable to the other groups described herein. That is, while terms such as "cycloalkyl" refer to both unsubstituted and substituted cycloalkyl moieties, further, substituted moieties may be specifically identified herein; for example, certain substituted cycloalkyl groups may be referred to as, for example, "alkylcycloalkyl". Similarly, a substituted alkoxy group may be specifically referred to as, for example, "haloalkoxy", and a specific substituted alkenyl group may be, for example, "alkenyl alcohol" and the like. Likewise, the use of a generic term (e.g., "cycloalkyl") and a specific term (e.g., "alkylcycloalkyl") does not imply that the generic term does not include the specific term.

The term "cycloalkyl" as used herein is a non-aromatic carbon-based ring consisting of at least three carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, and the like. The term "heterocycloalkyl" is a cycloalkyl group as defined above and is included within the meaning of the term "cycloalkyl" wherein at least one carbon atom of the ring is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus. Cycloalkyl and heterocycloalkyl groups may be substituted or unsubstituted. Cycloalkyl and heterocycloalkyl groups may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, nitrile, sulfonamide, or thiol as described herein.

The term "aryl" as used herein is a group containing any carbon-based aromatic group, including, but not limited to, benzene, naphthalene, phenyl, biphenyl, phenoxybenzene, and the like. The term "aryl" also includes heteroaryl, which is defined as a group containing an aromatic group having at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus. Likewise, the term "non-heteroaryl" is also included in the term "aryl" to define groups containing aryl groups that do not contain heteroatoms. The aryl group may be substituted or unsubstituted. The aryl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halogen, hydroxy, ketone, azido, nitro, silyl, sulfo-oxo, nitrile, sulfonamide, or thiol as described herein. The term "biaryl" is a specific type of aryl group, included in the definition of "aryl". Biaryl means that two aryl groups are joined together through a fused ring structure (as in naphthalene) or through one or more carbon-carbon bonds (as in biphenyl).

As used herein, the terms "halogen", "halide" and "halo" refer to the halogens fluorine, chlorine, bromine and iodine. It is also contemplated that, in various aspects, the halogen can be selected from fluorine, chlorine, bromine, and iodine. For example, the halogen may be selected from fluorine, chlorine and bromine. As another example, the halogen may be selected from fluorine and chlorine. As another example, the halogen may be selected from chlorine and bromine. As another example, the halogen may be selected from bromine and iodine. As another example, the halogen may be selected from chlorine, bromine, and iodine. In one aspect, the halogen can be fluorine. In a further aspect, the halogen can be chlorine. In a still further aspect, the halogen is bromine. In a still further aspect, the halogen is iodine.

It is also contemplated that in certain aspects, pseudohalogens (e.g., triflates, mesylates, tosylates, brosylates, etc.) may be used in place of halogens. For example, in certain aspects, a halogen may be substituted with a pseudohalogen. As another example, the pseudohalogen may be selected from triflate, mesylate, tosylate and brosylate. In one aspect, the pseudohalogen is triflate. In a further aspect, the pseudohalogen is mesylate. In a further aspect, the pseudohalogen is tosylate. In a further aspect, the pseudohalogen is p-bromobenzenesulfonate.

As used herein, the term "heterocycle" refers to monocyclic and polycyclic aromatic or non-aromatic ring systems in which at least one ring member is not carbon. Heterocycles include azetidine, dioxane, furan, imidazole, isothiazole, isoxazole, morpholine, oxazole (including 1,2, 3-oxadiazole, 1,2, 5-oxadiazole and 1,3, 4-oxadiazole), piperazine, piperidine, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, tetrahydrofuran, tetrahydropyran, tetrazine (including 1,2,4, 5-tetrazine), tetrazole (including 1,2,3, 4-tetrazole and 1,2,4, 5-tetrazole), thiadiazole (including 1,2, 3-thiadiazole, 1,2, 5-thiadiazole and 1,3, 4-thiadiazole), thiazole, thiophene, triazine (including 1,3, 5-triazine and 1,2, 4-triazine), triazole (including 1,2, 3-triazole, and thiadiazoles, 1,3, 4-triazole), and the like.

As used herein, the term "hydroxy" is represented by the formula — OH.

"R" as used herein¹”、“R²”、“R³”、“Rⁿ"(wherein n is an integer) may independently have one or more of the groups described above. For example, if R¹Is a straight chain alkyl group, one of the hydrogen atoms of the alkyl group may be optionally substituted with a hydroxyl group, an alkoxy group, an alkyl group, a halide, or the like. Depending on the group selected, the first group may be incorporated into the second group, or the first group may be pendant (i.e., attached) to the second group. For example, for the phrase "alkyl group comprising an amino group," the amino group may be incorporated within the backbone of the alkyl group. OrThus, the amino group may be attached to the backbone of the alkyl group. The nature of the group(s) selected will determine whether the first group is intercalated or attached to the second group.

As described herein, the compounds of the present invention may comprise an "optionally substituted" moiety. In general, the term "substituted", whether preceded by the term "optionally" or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a particular group, the substituents may be the same or different at each position. Combinations of substituents contemplated by the present invention are preferably those that result in the formation of stable or chemically feasible compounds. It is also contemplated that, in certain aspects, each substituent may be further optionally substituted (i.e., further substituted or unsubstituted), unless expressly stated to the contrary.

The compounds described herein may contain one or more double bonds, and thus may give rise to cis/trans (E/Z) isomers, as well as other conformational isomers. Unless stated to the contrary, the present invention includes all such possible isomers, as well as mixtures of such isomers.

Unless otherwise indicated, molecular formulas with chemical bonds shown only as solid lines rather than wedges or dashed lines contemplate each possible isomer (e.g., each enantiomer and diastereomer), as well as mixtures of isomers (e.g., racemic or partially racemic mixtures). The compounds described herein may contain one or more asymmetric centers and may therefore give rise to diastereomers and optical isomers. Unless indicated to the contrary, the present invention includes all such possible diastereomers as well as racemic mixtures thereof, substantially pure resolved enantiomers thereof, all possible geometric isomers thereof, and pharmaceutically acceptable salts thereof. Mixtures of stereoisomers are also included, as are isolated specific stereoisomers. During the synthetic procedures used to prepare these compounds, or during the use of racemization or epimerization procedures known to those skilled in the art, the products of these procedures can be mixtures of stereoisomers.

Many organic compounds exist in optically active forms that have the ability to rotate the plane of plane polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of a molecule with respect to its chiral center. The prefixes d and l or (+) and (-) are used to denote the rotation sign of a compound with respect to plane polarized light, and (-) or l denotes that the compound is left-handed. Compounds prefixed with (+) or d are dextrorotatory. For a given chemical structure, these compounds (called stereoisomers) are identical except that they are mirror images that are not superimposable on each other. A particular stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is commonly referred to as a mixture of enantiomers. A50: 50 mixture of enantiomers is referred to as a racemic mixture.

Many of the compounds described herein may have one or more chiral centers and thus may exist in different enantiomeric forms. If desired, the chiral carbons may be indicated by an asterisk. When the bond to the chiral carbon is depicted as a straight line in the disclosed formula, it is understood that the (R) and (S) configurations of the chiral carbon, and thus the enantiomers and mixtures thereof, are encompassed by the formula. As used in the art, when it is desired to specify an absolute configuration with respect to a chiral carbon, one bond of the chiral carbon may be depicted as a wedge (bonded to an atom above the plane) and the other bond may be depicted as a series or wedge of short parallel lines (bonded to an atom below the plane). The Cahn-Inglod-Prelog system can be used to assign either the (R) or (S) configuration to a chiral carbon.

The compounds described herein contain atoms in both natural isotopic and unnatural abundance. The disclosed compounds can be the same isotopically-labeled or isotopically-substituted compounds as described, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include hydrogen, carbon, nitrogen, oxygen, hydrogen,Isotopes of phosphorus, fluorine and chlorine, e.g. each²H、³H、¹³C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³⁵S、¹⁸F and³⁶and (4) Cl. The compounds further include prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labeled compounds of the present invention, for example, comprise a radioactive isotope (e.g.³H and¹⁴C) those compounds of (a), are useful in drug and/or stromal tissue distribution assays. Tritiated isotopes (i.e.³H) And carbon-14 isotopes (i.e.¹⁴C) They are particularly preferred for their ease of preparation and detectability. In addition, heavier isotopes are used (e.g. deuterium (i.e. deuterium)²H) Alternative) may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in some circumstances. Isotopically-labelled compounds of the present invention and prodrugs thereof can generally be prepared by: the non-isotopically labeled reagent is replaced by a readily available isotopically labeled reagent.

The compounds described in the present invention may exist as solvates. In some cases, the solvent used to prepare the solvate is an aqueous solution, and then the solvate is often referred to as a hydrate. The compounds may exist as hydrates, which may be obtained, for example, by crystallization from solvents or from aqueous solutions. In this regard, one, two, three or any number of solvate or water molecules may be combined with the compounds according to the present invention to form solvates and hydrates. Unless stated to the contrary, the present invention includes all such possible solvates.

It is also understood that certain compounds described herein may exist as equilibrium states of tautomers. For example, ketones having an alpha-hydrogen can exist in an equilibrium state of the keto form and the enol form.

Also, amides having N-hydrogen may exist in an equilibrium state of the amide form and the imide acid form. Unless stated to the contrary, the present invention includes all such possible tautomers.

It is known that chemical substances form solids that exist in different ordered states, which are called polymorphic forms (form) or variants (modification). Different variants of polymorphs can differ greatly in their physical properties. The compounds according to the invention may exist in different polymorphic forms, while particular variants may be metastable. Unless stated to the contrary, the present invention includes all such possible polymorphic forms.

In some aspects, the structure of a compound may be represented by the formula:

it is understood to be equivalent to the formula:

where n is typically an integer. That is, RⁿIs understood to represent five independent substituents: r^n(a)、R^n(b)、R^{n (c)}、R^n(d)、R^n(e). By "independent substituents" is meant that each R substituent may be independently defined. For example, if in one example, R^n(a)Is halogen, then in this case R^n(b)Not necessarily halogen.

Certain materials, compounds, compositions, and components disclosed herein can be obtained commercially or readily synthesized using techniques commonly known to those skilled in the art. For example, the starting materials and Reagents for preparing the disclosed Compounds and compositions can be obtained from commercial suppliers (e.g., Sigma-Aldrich chemical company (Milwaukee, Wis.), Across organics (NJ), Fisher Scientific (Pittsburgh, Pa.), or Sigma (St. Louis, Mo.),) or by methods known to those skilled in the art, according to the references (e.g., Fieser and materials Reagents for Organic Synthesis, Volumes 1-17(John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5and complexes (Elsevier Science Publishers, 1989); organics, Volumes 1-40(John trade company and company, Inc., and company's, Inc.; Wilcompany and company, Inc.; Wilcompany, Inc., and company, Inc.; Williams and company, Inc.; production company, Inc., and company, Inc., and company.

Unless expressly stated otherwise, it is in no way intended that any method set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not actually recite an order to be followed by its steps or it is not otherwise specifically stated in the claims or descriptions that the steps are to be limited to a specific order, it is not intended that an order be inferred, in any respect. This applies to any non-obvious basis for interpretation, including logic issues regarding step arrangement or operational flow; simple meaning derived from grammatical organization or punctuation; and the number or type of embodiments described in the specification.

Disclosed are the components used to prepare the compositions of the present invention as well as the compositions themselves to be used within the methods disclosed herein. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of various individual and collective combinations and permutations of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed and a number of modifications that can be made to a number of molecules including the compound are discussed, each and every combination and permutation of the compound and possible modifications are specifically contemplated unless specifically indicated to the contrary. Thus, if a class of molecules A, B and C is disclosed, as well as a class of molecules D, E and F, and an example of a combination molecule, A-D, is disclosed, then even if each is not individually recited, each is individually or collectively contemplated as meaning a combination, it is contemplated that A-E, A-F, B-D, B-E, B-F, C-D, C-E and C-F are disclosed. Likewise, any subset or combination of these is also disclosed. Thus, for example, it will be appreciated that subgroups of A-E, B-F and C-E are disclosed. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the compositions of the present invention. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the methods of the present invention.

It is understood that the compositions disclosed herein have certain functions. Certain structural requirements for performing the disclosed functions are disclosed herein, and it should be understood that there are various structures that can perform the same functions associated with the disclosed structures, and that these structures will generally achieve the same results.

B. Compound (I)

In one aspect, the present invention relates to compounds useful as ENPP1 inhibitors. Furthermore, in one aspect, the compounds of the invention are useful for treating disorders of uncontrolled cellular proliferation. In a further aspect, the disorder of uncontrolled cellular proliferation is a cancer or a tumor. In a still further aspect, the disorder of uncontrolled cellular proliferation is associated with ENPP1 dysfunction, as further described herein.

In another aspect, the compounds of the invention are useful for the treatment of diseases of bacterial or viral origin. Accordingly, in one aspect, the present invention provides a method of treating a disease caused by a bacterium or virus, comprising administering to a subject a therapeutically effective amount of a compound of the present invention.

It is contemplated that each of the disclosed derivatives may be optionally further substituted. It is also contemplated that any one or more derivatives may optionally be omitted from the present invention. It is to be understood that the disclosed compounds can be provided by the disclosed methods. It is also to be understood that the disclosed compounds can be used in the disclosed methods of use.

1. Structure of the product

In one aspect, the invention relates to compounds of formula I:

wherein

Y is S;

z is-H or-N;

X₁is-C ═ O or-NH,

X₂is-C ═ O or-NH, with the proviso that if X₁is-C ═ O, then X₂is-NH; and if X₁is-NH, then X₂is-C ═ O;

R₁selected from the group consisting of: -H, halogen, -OCF₃、-CF₃、-CN、OCH₃and-CH₃；

R₂Selected from the group consisting of: -H, -NH₂-OH and-CH₃；

R₃Selected from the group consisting of: -H, -NH₂-OH and-CH₃；

R₄Selected from the group consisting of: -H, -CH₂-CH₃and-CH₃；

R₅Selected from the group consisting of:

and is

R₆Selected from the group consisting of: -OCH₃、OH、F、Cl、-CH₃、O-CH₂-CH₃And OCH₂-CF₃、OCHF₂，

Or an isomer thereof or a pharmaceutically acceptable salt thereof.

In a preferred embodiment of the process according to the invention,

z is-N;

X₁is-C ═ O;

X₂is-NH;

R₁selected from the group consisting of: halogen and-CN;

R₂selected from the group consisting of: -H and-CH₃；

R₃is-H;

R₄is-H;

R₅is that

And is

R₆Selected from the group consisting of: -OCH₃And OH.

In another preferred embodiment, R₁Selected from the group consisting of: -Cl and-CN.

Examples of compounds provided include:

the invention also provides pharmaceutical compositions comprising a therapeutically effective amount of any of the compounds of the invention and a pharmaceutically acceptable carrier.

The invention also provides a method for treating a disorder of uncontrolled cellular proliferation in a mammal, the method comprising the step of administering to the mammal an effective amount of any of the compounds of the invention.

The present invention also provides a method of reducing ENPP1 activity in a mammal, the method comprising the step of administering to the mammal an effective amount of any of the compounds of the present invention.

The present invention also provides a method of inhibiting ENPP1 activity in a mammal, comprising the step of administering to the mammal an effective amount of any of the compounds of the present invention.

C. Process for preparing compounds

In one aspect, the present invention relates to methods of making compounds useful as ENPP1 inhibitors. In a further aspect, the product of the disclosed preparation method is a modulator of ENPP1 activity.

The compounds of the invention can be prepared by using the reactions shown in the following schemes, in addition to other standard procedures known in the literature, which are illustrated in the experimental section or are clear to the skilled person. For clarity, examples having a single substituent are shown, where multiple substituents are allowed under the definitions disclosed herein.

The reactions used to produce the compounds of the present invention are prepared by using reactions as shown in the following reaction schemes, in addition to other standard procedures known in the literature or known to those skilled in the art. The following examples are provided for a more complete understanding of the present invention and are intended to be illustrative only and should not be construed as limiting.

In one aspect, the disclosed compounds include the products of the synthetic methods described herein. In a further aspect, the disclosed compounds include compounds produced by the synthetic methods described herein. In yet a further aspect, the invention includes a pharmaceutical composition comprising a therapeutically effective amount of the product of the disclosed methods and a pharmaceutically acceptable carrier. In a still further aspect, the invention includes a method for making a medicament comprising combining at least one compound of any disclosed compound or at least one product of the disclosed method with a pharmaceutically acceptable carrier or diluent.

It is believed that one skilled in the art can determine the reaction conditions and amounts of ingredients without specifying them. It is contemplated that each disclosed method may also include additional steps, acts, and/or components. It is also contemplated that any one or more steps, operations, and/or components may optionally be omitted from the present invention. It is understood that the disclosed methods can be used to provide the disclosed compounds. It is also understood that the products of the disclosed methods can be used in the disclosed methods of use.

D. Pharmaceutical composition

In one aspect, the invention relates to pharmaceutical compositions comprising the disclosed compounds. That is, a pharmaceutical composition can be provided that comprises a therapeutically effective amount of at least one disclosed compound or at least one product of the disclosed methods and a pharmaceutically acceptable carrier.

In a further aspect, the invention relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of the product of the disclosed synthetic process. In a further aspect, the effective amount is a therapeutically effective amount. In a further aspect, the effective amount is a prophylactically effective amount. In a further aspect, the compound is a disclosed compound.

In certain aspects, the disclosed pharmaceutical compositions comprise the disclosed compounds (including pharmaceutically acceptable salts thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally other therapeutic ingredients or adjuvants. The compositions of the invention include those suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and the nature and severity of the conditions to which the active ingredient is being administered. The pharmaceutical compositions may conveniently be presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

The term "pharmaceutically acceptable salt" as used herein refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When the compounds of the present invention are acidic, their corresponding salts may be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (both higher (-ic) and lower (-ous)), ferric, ferrous, lithium, magnesium, manganese (both higher and lower), potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, as well as salts of cyclic amines and substituted amines (e.g., naturally occurring and synthetic substituted amines). Other pharmaceutically acceptable organic non-toxic bases from which salts can be formed include ion exchange resins such as arginine, betaine, caffeine, choline, N' -dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

As used herein, the term "pharmaceutically acceptable non-toxic acid" includes inorganic acids, organic acids and salts prepared therefrom, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroxyethyl, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, polyamic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid and the like. Preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric and tartaric acids.

In fact, the compounds of the present invention or pharmaceutically acceptable salts thereof may be intimately admixed with pharmaceutical carriers as the active ingredient according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present invention may be presented as discrete units suitable for oral administration, such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient. Furthermore, the composition may be present as a powder, as a particle, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion or as a water-in-oil liquid emulsion. In addition to the above-mentioned conventional dosage forms, the compounds of the present invention and/or pharmaceutically acceptable salts thereof may also be administered by a controlled release route and/or delivery device. The compositions may be prepared by any pharmaceutical method. Generally, such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more required ingredients. Generally, compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped to the desired appearance.

Accordingly, the pharmaceutical compositions of the present invention may include a pharmaceutically acceptable carrier and a compound of the present invention or a pharmaceutically acceptable salt thereof. The compounds of the present invention or pharmaceutically acceptable salts thereof may also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.

The pharmaceutical carrier employed may be, for example, a solid, liquid or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate and stearic acid. Examples of liquid carriers are syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

In preparing the compositions for oral dosage form, any convenient pharmaceutical medium may be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like may be used to form oral liquid preparations (e.g., suspensions, elixirs, and solutions); and carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like may be used to form oral solid dosage forms (e.g., powders, capsules and tablets). Tablets and capsules are the preferred oral dosage units due to ease of administration, whereby solid pharmaceutical carriers are employed. Optionally, the tablets may be coated by standard aqueous or non-aqueous techniques.

Tablets containing the composition of the invention may be prepared by compression or moulding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

The pharmaceutical compositions of the invention comprise a compound of the invention (or a pharmaceutically acceptable salt thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally one or more other therapeutic agents or adjuvants. The compositions of the invention include compositions suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and the nature and severity of the conditions for which the active ingredient is administered. The pharmaceutical compositions may conveniently be presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

Pharmaceutical compositions of the invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compound in water. Suitable surfactants, such as hydroxypropyl cellulose, may be included. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. In addition, preservatives may be included to prevent undesirable microbial growth.

Pharmaceutical compositions of the invention suitable for injectable use include sterile aqueous solutions or dispersions. In addition, the compositions may be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In all cases, the final injectable form must be sterile and must be an effective liquid for easy injection. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; therefore, preferably, the contaminating action of microorganisms such as bacteria and fungi should be prevented. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

The pharmaceutical compositions of the present invention may be in a form suitable for topical use, for example, aerosols, creams, ointments, lotions, dusting powders, mouthwashes, gargles, and the like. In addition, the composition may be in a form suitable for use in a transdermal device. These formulations can be prepared by conventional processing methods using the compounds of the present invention or pharmaceutically acceptable salts thereof. For example, a cream or ointment is prepared by mixing a hydrophilic material and water with about 5% to about 10% by weight of the compound to produce a cream or ointment having a desired consistency.

The pharmaceutical compositions of the present invention may be in a form suitable for rectal administration wherein the carrier is a solid. Preferably, the mixture forms a unit dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be conveniently formed by first mixing the composition with the softened or melted carrier, followed by cooling and shaping in a mould.

In addition to the above-mentioned carrier ingredients, the above-mentioned pharmaceutical preparations may suitably comprise one or more additional carrier ingredients, such as diluents, buffers, flavouring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants) and the like. In addition, other adjuvants may be included to render the formulation isotonic with the blood of the intended recipient. Compositions containing the compounds of the present invention and/or pharmaceutically acceptable salts thereof may also be prepared in the form of a powder or liquid concentrate.

Suitable dosage levels under therapeutic conditions where inhibition or down-regulation of ENPP1 protein activity is desired are typically from about 0.01 to 500mg/kg patient body weight per day, and may be administered in single or multiple doses. Preferably, the dosage level is from about 0.1 to about 250mg/kg per day; more preferably 0.5 to 100mg/kg per day. Suitable dosage levels may be about 0.01 to 250mg/kg per day, about 0.05 to 100mg/kg per day, or about 0.1 to 50mg/kg per day. Within this range, the dose may be 0.05 to 0.5, 0.5 to 5.0, or 5.0 to 50mg/kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000mg of active ingredient, in particular 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000mg of active ingredient, for the symptomatic adjustment of the dosage to the patient to be treated. The compound may be administered from 1 to 4 times per day, preferably once or twice per day. The administration regimen may be adjusted to provide the optimal therapeutic response.

It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors. These factors include the age, weight, general health, sex, and diet of the patient. Other factors include the time and route of administration, rate of excretion, drug combination, and the type and severity of the particular disease being treated.

The present invention also relates to a method of making a medicament for inhibiting or down-regulating ENPP1 protein activity (e.g., treating a disorder of uncontrolled cellular proliferation, or one or more neurodegenerative disorders associated with ENPP1 dysfunction) in a mammal (e.g., a human), comprising combining one or more of the disclosed compounds, products, or compositions with a pharmaceutically acceptable carrier or diluent. Thus, in one aspect, the invention relates to a process for the manufacture of a medicament comprising combining at least one disclosed compound or at least one disclosed product with a pharmaceutically acceptable carrier or diluent.

The disclosed pharmaceutical compositions may further comprise other therapeutically active compounds, which are generally applied in the treatment of the above mentioned pathological conditions.

It is understood that the disclosed compositions can be prepared from the disclosed compounds. It is also to be understood that the disclosed compositions can be used in the disclosed methods of use.

E. Methods of using compounds and compositions

The disclosed compounds can be used as single agents or in combination with one or more other drugs for the treatment, prevention, control, amelioration, or reduction of risk of the above-mentioned diseases, disorders, and conditions for which compounds of formula I or other drugs have utility, where the drugs in combination are safer or more effective than either drug alone. Other drugs can be administered either simultaneously or sequentially with the disclosed compounds by the routes and amounts commonly used. When the disclosed compounds are used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compounds is preferred. However, the combination therapy may also be administered in an overlapping regimen. It is also contemplated that the combination of one or more active ingredients and the disclosed compounds will be more effective than either as a single dose.

The pharmaceutical compositions and methods of the present invention may further comprise other therapeutically active compounds as described herein, which are typically employed in the treatment of the above-mentioned pathological conditions.

1. Method of treatment

The compounds disclosed herein are useful for treating, preventing, ameliorating, controlling, or reducing the risk of a variety of disorders in which a patient or subject would benefit from inhibition or negative modulation of ENPP 1. In one aspect, a method of treating or preventing a disorder in a subject is provided, comprising administering to the subject at least one disclosed compound in a dose and amount effective to treat the disorder in the subject; at least one disclosed pharmaceutical composition; and/or at least one of the disclosed products.

Also provided is a method of treating one or more conditions for which inhibition of ENPP1 is expected to be beneficial in a subject, comprising administering to the subject at least one disclosed compound in a dose and amount effective to treat the condition in the subject; at least one disclosed pharmaceutical composition; and/or at least one of the disclosed products.

In one aspect, a method of treating an uncontrolled cell proliferation disorder is provided, comprising: administering at least one disclosed compound to a subject at a dose and amount effective to treat the disorder in the subject; at least one disclosed pharmaceutical composition; and/or at least one of the disclosed products. In a further aspect, there is provided a method of treating or preventing a neurodegenerative disorder comprising: administering at least one disclosed compound to a subject at a dose and amount effective to treat the disorder in the subject; at least one disclosed pharmaceutical composition; and/or at least one of the disclosed products. Also provided are methods for treating a condition in a mammal comprising the step of administering to the mammal at least one disclosed compound, composition or medicament.

The present invention relates to the use of said chemical composition for the treatment of a disease or disorder in a patient, preferably a human, wherein inhibition of ENPP1 is expected to have a therapeutic effect, such as disorders of uncontrolled cellular proliferation (e.g. cancer) and neurodegenerative disorders (e.g. alzheimer's, huntington's and parkinson's), diseases caused by bacteria and/or viruses, by administering one or more of the disclosed compounds or products.

The compounds of the invention are also useful in immunotherapy. In one embodiment, the compounds of the invention are used to treat uncontrolled cell proliferation disorders and/or diseases caused by bacteria and/or viruses by immunotherapy, which means that the compounds elicit an immunotherapeutic response to treat these diseases.

The compounds disclosed herein are useful for treating, preventing, ameliorating, controlling, or reducing the risk of various uncontrolled cell proliferation disorders.

Methods of using the disclosed compounds, compositions, or medicaments are also provided. In one aspect, the method of use relates to the treatment of a disorder. In a further aspect, the disclosed compounds can be used as a single agent or in combination with one or more other drugs for treating, preventing, managing, ameliorating or reducing the risk of the aforementioned diseases, disorders and conditions for which the compounds or other drugs have utility, wherein the drugs in combination are safer or more effective than either drug alone. Other drugs can be administered either simultaneously or sequentially with the disclosed compounds by the routes and amounts commonly used. When the disclosed compounds are used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compounds is preferred. However, the combination therapy may also be administered in an overlapping regimen. It is also contemplated that a combination of one or more active ingredients and the disclosed compounds may be more effective than as a single dose.

Examples of disorders that can be treated with the provided compounds include disorders of uncontrolled cellular proliferation. In a still further aspect, the disorder of uncontrolled cellular proliferation is cancer. In a still further aspect, the cancer is leukemia. In a further aspect, the cancer is a sarcoma. In a still further aspect, the cancer is a solid tumor. In a still further aspect, the cancer is lymphoma.

It is understood that "cancer" refers to or describes a physiological condition in mammals that is generally characterized by unregulated cell growth. The cancer may be multidrug resistant (MDR) or drug sensitive. Examples of cancer include, but are not limited to, carcinoma (carcinoma), lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include breast, prostate, colon, squamous cell, small-cell lung, non-small cell lung, gastrointestinal, pancreatic, cervical, ovarian, peritoneal, liver (e.g., liver, bladder, colorectal, endometrial), kidney, and thyroid cancers.

In various aspects, further examples of cancer are basal cell carcinoma, biliary tract carcinoma; bone cancer; brain and CNS cancers; choriocarcinoma; connective tissue cancer; esophageal cancer; eye cancer; head and neck cancer; gastric cancer; intraepithelial tumors; laryngeal cancer; lymphomas (including hodgkin lymphoma and non-hodgkin lymphoma); melanoma; a myeloma cell; neuroblastoma; oral cancer (e.g., lip, tongue, mouth, and pharynx); retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; a sarcoma; skin cancer; gastric cancer; testicular cancer; uterine cancer; urinary cancers as well as other cancers and sarcomas.

In a further aspect, the cancer is a hematologic cancer. In a further aspect, the hematologic cancer is selected from Acute Myelogenous Leukemia (AML), Acute Lymphocytic Leukemia (ALL), Chronic Myelogenous Leukemia (CML), Chronic Lymphocytic Leukemia (CLL), hairy cell leukemia, chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia (JMML), hodgkin lymphoma, non-hodgkin lymphoma, multiple myeloma, single myeloma, local myeloma, and extramedullary myeloma. In a further aspect, the cancer is selected from chronic lymphocytic leukemia, small lymphocytic lymphoma, B-cell non-hodgkin's lymphoma, and large B-cell lymphoma.

In a further aspect, the cancer is a brain cancer. In a further aspect, the brain cancer is selected from the group consisting of glioma, medulloblastoma, primary neuroectodermal tumor (PNET), acoustic neuroma, glioma, meningioma, pituitary adenoma, schwannoma, CNS lymphoma, primary neuroectodermal tumor, craniopharyngioma, chordoma, medulloblastoma, brain neuroblastoma, central neuroblastoma, pinealoblastoma, atypical teratoid rhabdoid tumor, chondrosarcoma, choroid plexus cancer, choroid plexus papilloma, craniopharyngioma, neuroepithelial tumor with dysplasia, ganglioneuroma, germ cell tumor, hemangioblastoma, vascular endothelial cell tumor, and metastatic brain tumor. In a still further aspect, the glioma is selected from the group consisting of ependymoma, astrocytoma, oligodendroglioma, and oligodendroastrocytoma. In a further aspect, the glioma is selected from the group consisting of juvenile hairy cell astrocytoma, sub-ependymal giant cell astrocytoma, ganglion cell glioma, sub-ependymal tumor, pleomorphic yellow astrocytoma, anaplastic astrocytoma, glioblastoma multiforme, brain stem glioma, oligodendroglioma, ependymal tumor, oligodendroastrocytoma, cerebellar astrocytoma, infantile profibroproliferative astrocytoma (desmoplastic in fantille astrocytoma), sub-ependymal giant cell astrocytoma, diffuse astrocytoma, mixed glioma, optic glioma, brain glioma disease, multifocal glioma, multicentric glioblastoma multiforme, paraganglioma, and ganglion cell tumor.

In one aspect, the cancer may be a cancer selected from the group consisting of blood, brain, genitourinary tract, gastrointestinal tract, colon, rectum, breast, kidney, lymphatic system, stomach, lung, pancreas, and skin. In a further aspect, the cancer is selected from the group consisting of prostate cancer, glioblastoma multiforme, endometrial cancer, breast cancer, and colon cancer. In a further aspect, the cancer is selected from the group consisting of breast, ovarian, prostate, head, neck and kidney cancers. In a still further aspect, the cancer is selected from cancers of the blood, brain, genitourinary tract, gastrointestinal tract, colon, rectum, breast, liver, kidney, lymphatic system, stomach, lung, pancreas, and skin. In a still further aspect, the cancer is selected from lung and liver cancer. In a still further aspect, the cancer is selected from the group consisting of breast, ovarian, testicular, and prostate cancer. In a still further aspect, the cancer is a cancer of the breast. In a still further aspect, the cancer is a cancer of the ovary. In a further aspect, the cancer is a cancer of the prostate. In a still further aspect, the cancer is cancer of the testis.

In various aspects, the disorder associated with dysfunction of ENPP1 comprises a neurodegenerative disorder. In a further aspect, the neurodegenerative disease is selected from alzheimer's disease, parkinson's disease, and huntington's disease.

The compounds may also be used in methods of preventing, treating, controlling, ameliorating, or reducing the risk of diseases, disorders, and conditions described herein. The compounds may also be used in combination with other agents in methods of preventing, treating, controlling, ameliorating, or reducing the risk of the foregoing diseases, disorders, and conditions.

The invention also relates to the administration of an ENPP1 inhibitor for improving the therapeutic outcome in the case of uncontrolled cell proliferation disorders, including cancer. That is, in one aspect, the present invention relates to a combination therapy (cotherapeutic) method comprising the step of administering to a mammal an effective amount and dose of at least one compound of the present invention that is associated with cancer therapy.

In a further aspect, administration improves the therapeutic outcome of the cancer therapy. Administration in connection with cancer therapy may be continuous or intermittent. Administration need not be simultaneous with therapy, and can be before, during, and/or after therapy. For example, cancer therapy can be provided within 1,2,3,4, 5,6, 7 days before or after administration of the compound. As another example, cancer therapy can be provided within 1,2,3, or 4 weeks before or after administration of the compound. As yet another example, cognitive or behavioral therapy may be provided within a period of 1,2,3,4, 5,6, 7, 8, 9, or 10 half-lives of the administered compound, either before or after administration.

In one aspect, the disclosed compounds can be used in combination with one or more other drugs to treat, prevent, control, ameliorate, or reduce the risk of a disease or condition for which the disclosed compounds or other drugs can have utility, where the drugs in combination are safer or more effective than either drug alone. Such other agents may be administered, by a route and in an amount commonly used therefor, either simultaneously or sequentially with a compound of the present invention. When a compound of the present invention is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the disclosed compound is preferred. However, combination therapy may also include therapies in which the disclosed compound and one or more other drugs are administered in different overlapping regimens. It is also contemplated that the disclosed compounds and other active ingredients, when used in combination with one or more other active ingredients, may be used in lower doses than when used alone.

Accordingly, pharmaceutical compositions include those that contain one or more other active ingredients in addition to a compound of the invention.

Such combinations include not only combinations of the disclosed compounds with one other active compound, but also combinations of the disclosed compounds with two or more other active compounds. Likewise, the disclosed compounds can be used in combination with other drugs that are useful in the prevention, treatment, control, amelioration, or reduction of risk of diseases or conditions for which the disclosed compounds are useful. Such other drugs may be administered, by a route and in an amount commonly used therefor, simultaneously or sequentially with the compound of the present invention. When a compound of the present invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the disclosed compounds is preferred. Accordingly, pharmaceutical compositions also include those that contain one or more other active ingredients in addition to a compound of the present invention.

The weight ratio of the disclosed compound to the second active ingredient can vary and will depend on the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when a compound of the present invention is combined with another agent, the weight ratio of the disclosed compound to the other agent is generally in the range of from about 1000: 1 to about 1: 1000, preferably in the range of from about 200: 1 to about 1: 200. Combinations of the compounds of the invention and other active ingredients will generally also be within the above-described ranges, but in each case an effective dose of each active ingredient should be used.

In such combinations, the disclosed compounds and other active agents may be administered alone or in combination (inconnection). In addition, one ingredient may be administered before, simultaneously with, or after the other agent.

Thus, the compounds of the present invention may be used alone or in combination with other agents known to be beneficial in the present indications or other drugs that affect receptors or enzymes that increase the efficacy, safety, convenience, or reduce undesirable side effects or toxicity of the disclosed compounds. The compounds of the present invention and other agents may be co-administered in concomitant therapy (comomatant therapy) or in a fixed combination.

In one aspect, the compounds may be used in combination with anti-cancer therapeutics or other known therapeutics.

In the treatment of conditions where inhibition or down-regulation of ENPP1 is desired, suitable dosage levels will generally be from about 0.01 to 1000mg/kg of patient body weight per day, which may be administered in single or multiple doses. Preferably, the dosage level is from about 0.1 to about 250mg/kg per day; more preferably from about 0.5 to 100mg/kg per day. Suitable dosage levels may be about 0.01 to 250mg/kg per day, about 0.05 to 100mg/kg per day, or about 0.1 to 50mg/kg per day. Within this range, the dose may be 0.05 to 0.5, 0.5 to 5, or 5 to 50mg/kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000mg of active ingredient, in particular 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000mg of active ingredient, for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered from 1 to 4 times per day, preferably once or twice per day. The dosage regimen may be adjusted to provide the optimal therapeutic response. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

Thus, in one aspect, the present invention relates to a method for inhibiting or down-regulating ENPP1 in at least one cell, said method comprising the step of contacting the at least one cell with at least one compound of the present invention in an amount effective to modulate or activate, for example, an ENPP1 activity response in the at least one cell. In a further aspect, the cell is mammalian, e.g., human. In a further aspect, the cell has been isolated from the subject prior to the contacting step. In a further aspect, the contacting is by administration to the subject.

a. Treatment of uncontrolled cell proliferation disorders

In one aspect, the invention relates to a method of treating a disorder of uncontrolled cellular proliferation in a mammal, the method comprising the step of administering to the mammal an effective amount of at least one disclosed compound or a product of a disclosed method of making a compound, or a pharmaceutically acceptable salt, hydrate, solvate, or polymorph thereof, thereby treating the disorder of uncontrolled cellular proliferation.

In a still further aspect, the effective amount is a therapeutically effective amount. In a still further aspect, the effective amount is a prophylactically effective amount.

In a further aspect, the mammal is a human. In a still further aspect, the method further comprises the step of identifying a mammal in need of treatment for a disorder of uncontrolled cellular proliferation. In a still further aspect, the mammal has been diagnosed with a need for treatment of a condition of uncontrolled cellular proliferation prior to the administering step.

In a further aspect, the disorder of uncontrolled cellular proliferation is cancer. In a still further aspect, the cancer is leukemia. In a further aspect, the cancer is a sarcoma. In a still further aspect, the cancer is a solid tumor. In a still further aspect, the cancer is lymphoma. In a further aspect, the cancer is selected from chronic lymphocytic leukemia, small lymphocytic lymphoma, B-cell non-hodgkin's lymphoma, and large B-cell lymphoma. In a still further aspect, the cancer is selected from cancers of the blood, brain, genitourinary tract, gastrointestinal tract, colon, rectum, breast, liver, kidney, lymphatic system, stomach, lung, pancreas, and skin. In a still further aspect, the cancer is selected from lung and liver cancer. In a still further aspect, the cancer is selected from the group consisting of breast, ovarian, testicular, and prostate cancer. In a still further aspect, the cancer is a cancer of the breast. In a still further aspect, the cancer is a cancer of the ovary. In a further aspect, the cancer is a cancer of the prostate. In a still further aspect, the cancer is cancer of the testis.