(e) -n '- (1-phenylethylidene) benzohydrazide analogs substituted as histone demethylase inhibit

专利摘要:
Substituted (e) -n '- (1-phenylethylidene) benzohydrazide analogues as histone demethylase inhibitors The present invention relates to substituted (e) -n' - (1-phenylethylidene) benzohydrazide analogues or (3- (5-chloro -2-hydroxyphenyl-4-methyl-1H-pyrazol-1-yl) (3- (morpholinosulfonyl) phenyl) methanone, derivatives thereof, and related compounds which are useful as lysine-specific histone demethylase inhibitors, including lsd1. compounds for preparing the compounds, pharmaceutical compositions comprising the compounds, and methods for using the compounds and compositions for treating disorders associated with lysine-specific demethylase dysfunction, a pharmaceutically acceptable salt, hydrate, solvate, or polymorph thereof, and one or more of: (a) at least one agent known to enhance histone demethylase activity, (b) at least one agent known to reduce histone demethylase activity, (c) at least one known agent treating an uncontrolled cell proliferation disorder; (d) at least one agent known to treat a neurodegenerative disorder; (e) instructions for treating a neurodegenerative disorder, or (f) instructions for treating a disorder associated with uncontrolled cell proliferation.
公开号:BR112015032113B1
申请号:R112015032113-5
申请日:2014-06-19
公开日:2019-01-29
发明作者:Hariprasad Vankayalapati；Venkataswamy Sorna；Steve L. Warner；David J. Bearss；Sunil Sharma；Bret Stephens
申请人:University Of Utah Research Foundation；
IPC主号:

专利说明:

Patent Descriptive Report for: "(E) -Ν '- (1 — PHENILETILIDEN) BENZOHYDRAZIDE ANALOGS REPLACED AS HYDON DEMETILASE INHIBITORS" BACKGROUND OF THE INVENTION
In the last decade, it has become clear that epigenetic changes, which alter gene activity without altering the DNA sequence, have collaborated with genetic errors to promote cancer development and progression (Tsai, HC and Baylin, SB Cell Res 2011, 21 (3), 502-17, and Fullgrabe, J., Kavanagh, E., and Joseph, B. Oncogene 2011). Regulation of DNA and protein modifications associated with DNA has become an area of intense interest, and the enzymes involved in these processes have been suggested as a new class of protein targets for drug development. The main proteins associated with DNA are histone proteins. Histone tails are subject to a variety of post-translational modifications, such as phosphorylation, acetylation, methylation and ubiquitination, and such modifications, especially acetylation and methylation in lysine residues, play an important role in regulating gene expression and are often unregulated. in cancer (Fullgrabe, J., Kavanagh, E. and Joseph, B. Oncogene 2011).
Recently, an enzyme called specific lysine demethylase 1 (LSD1) has been found to catalyze oxidative demethylation of monomethylated and dimethylated histone H3 to lysine 4 (H3K4mel and H3K4me2) and to lysine 9 (H3K9mel and H3K9me2). flavin and adenine dinucleotide dependent reaction (FAD) (Shi, Y., et al.
Cell 2004, 119 (7), 941-53; and Metzger, E., et al. Nature 2005, 437 (7057), 436-9), Given that histone acetylation is associated with loose chromatin and gene activation, histone methylation is less direct. Using LSD1-regulated lysine residues as an example, H3K4 methylation is generally associated with gene activation, whereas H3K9 methylation is associated with repression of transcription.
There is currently a known mammalian homologue of LSD1 which is a protein designated in various ways such as LSD2, KDMlb and AOF1. It shares a similar domain homology, but has less than 31% sequence identity (Fang, R. et al. Molecular Cell 2010, 39: 222-233). LSD2 has been shown to be an H3K4mel / 2 demethylase that specifically regulates histone H3K4 methylation in the intragenic regions of its target genes (ibid.). Both LSD1 and LSD2 contain a SWIRM domain, a FAD coenzyme binding moiety and a C-terminal amine oxidase domain, all of which are essential for enzymatic activity. However, unlike LSD1, the LSD2 protein contains a CW zinc finger domain in its N-terminal domain, a region that is unstructured in LSD1. Also, LSD2 does not contain the LSD1 tower domain. At the cellular level, it has been suggested that LSD2 has a function in regulating transcription (ibid.). As expected, LSD2 also appears to play a role in regulating DNA methylation as well, although the function in DNA methylation may be specific to the developmental stage (ibid .; Ciccone, DN, et al. Nature 2009 461: 415 -418; Karytinos, A., et al., J. Biol. Chem. 2009 284: 17775-17782; and Yang, Z., et al., Cell Res. 2010 20: 276-287).
Several lines of evidence point to LSD1 as a possible therapeutic target in cancer. LSD1 is reportedly overexpressed in a variety of tumors, including ER negative breast, bladder, lung, and colorectal tumors neuroblastoma (Schulte, JH, et al. Cancer Res 2009, 69 (5), 2065-71; Lim, S., et al.
Carcinogenesis 2010, 31 (3), 512-20; and Hayami, S., et al.
Int J Cancer 2011, 128 (3), 574-86). Increased methylation of the permissive H3K4 tag by inhibiting LSD1 has been shown to reactivate tumor suppressor gene expression in cancer models (Huang, Y., et al. Clin Cancer Res 2009, 15 (23), 7217-28). In addition, LSD1 has been found to be associated with estrogen and androgen receptors that lead to repressive H3K9-specific demethylation, thereby increasing target gene expression (Metzger, E., et al. Nature 2005, 437 (7057)). 436-9, and Garcia-Bassets, I., et al., Cell 2007, 128 (3), 505-18). Thus, depending on LSD1-linked cofactors, LSD1 demethylation may contribute to cancer through the permissive H3K4 and repressive H3K9 mark. Therefore, LSD1 inhibition may be an effective strategy for re-expression of epigenetically silenced tumor suppressor genes, as well as for the unregulation of important cancer pathways in various cancers. Several LSD1 inhibitors have been reported, but they have shown poor selectivity and / or pharmacological properties, further complicating the exploration of LSD1 biology.
Monoamine oxidase (MAO) inhibitors, such as tranylcypromine and pargillin, have been reported as inhibitors of LSD1, and there are several reports of attempts to find derivatives with higher selectivity for LSD1 over MAO (Mimasu, S., et Biochemistry 2010, 49 (30), 6494-503; Binda, C., et al., J Am Chem Soc 2010, 132 (19), 6827-33; Culhane, JC, et al., J Am Chem Soc 2006, 128 (14), 4536-7; Culhane, JC, et al., J Am Chem Soc 2010, 132 (9), 3164-76; and Ueda, R., et al., J Am Chem Soc 2009, 131 (48) , 17536-7). These compounds irreversibly inactivate LSD1 by covalent binding to the FAD cofactor. Polyamine derivatives have also been evaluated as inhibitors of LSD1, where compounds with activity in the μ na range have been described (Huang, Y., et al. Clin Cancer Res 2009, 15 (23), 7217-28; Sharma, SK, et al J Med Chem 2010, 53 (14), 5197-212, and Huang, Y. et al Proc Natl Acad Sci USA 2007, 104 (19), 8023-8). In general, these and other reported LSD1 inhibitors are neither adequately selective nor sufficiently potent to optimally interact with the crucial amino acid residues of the substrate binding site present in LSD1.
In summary, LSD proteins play a key role in transcriptional and epigenetic regulation, and they are often altered in mammalian cancers, making them an attractive target for therapeutic intervention. Despite advances in drug discovery aimed at identifying inhibitors of LSD1 and / or LSD2 protein activity, there is still a shortage of compounds that are potent, effective and selective inhibitors of LSD1 or LSD2. In addition, there is a shortage of compounds effective in the treatment of cancer and other disorders associated with LSD1 and / or LSD2 dysfunction. These and other needs are met by the present invention.
SUMMARY
According to the purpose (s) of the invention as set forth and widely described herein, the invention in one aspect relates to compounds useful as inhibitors of lysine demethylase-specific demethylase or LSD. In another aspect, the disclosed compounds and products of the disclosed manufacturing methods, or a pharmaceutically acceptable salt, hydrate, solvate or polymorph thereof, are modulators of LSD activity, methods of doing so, pharmaceutical compositions comprising the same and methods of treating disorders associated with an LSD activity disorder using the same. In yet another further aspect, the present invention relates to compounds that bind to an LSD protein and negatively modulate LSD activity. The disclosed compounds may, in one aspect, exhibit subtype selectivity. In another aspect, the disclosed compounds exhibit selectivity for the LSD1 member of the LSD protein family. In yet another aspect, the disclosed compounds exhibit selectivity for the LSD2 member of the LSD protein family.
Also disclosed are pharmaceutical compositions comprising a therapeutically effective amount of a disclosed compound and a pharmaceutically acceptable carrier.
Synthetic methods for making the disclosed compounds are also disclosed. In another aspect, the products of the disclosed synthetic methods are disclosed.
Methods are disclosed for treating a disease associated with a dysfunction of LSD activity in a mammal which comprises the step of administering to the mammal a therapeutically effective amount of a disclosed compound, or a hydrate, solvate, polymorph or salt pharmaceutically. acceptable of it.
Also disclosed are methods for inhibiting LSD 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 hydrate, solvate, polymorph or salt thereof.
Also disclosed are methods for inhibiting LSD activity in at least one cell, comprising the activity of contacting at least one cell with an effective amount of at least one disclosed compound, or a hydrate, solvate, polymorph or salt. pharmaceutically acceptable thereof.
Uses of a disclosed compound or of a pharmaceutically acceptable hydrate, solvate, polymorph or salt thereof are also disclosed. The invention relates to pharmaceutical compositions comprising a pharmaceutically acceptable carrier and an effective amount of a disclosed compound, or a hydrate, solvate, polymorph or pharmaceutically acceptable salt thereof.
Also disclosed are kits comprising at least one disclosed compound, or a pharmaceutically acceptable hydrate, solvate, polymorph or salt thereof, and one or more of: (a) at least one agent known to enhance histone demethylase activity; (b) at least one agent known to decrease histone demethylase activity; (c) at least one agent known to treat an uncontrolled cell proliferation disorder; (d) at least one agent known to treat a neurodegenerative disorder; (e) instructions for treating a neurodegenerative disorder; or (f) instructions for treating a disorder associated with uncontrolled cell proliferation.
Also disclosed are methods 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. In another aspect, the invention relates to the use of a compound disclosed in the manufacture of a medicament for the treatment of a disorder associated with a dysfunction of LSD activity. In an additional aspect, LSD activity dysfunction is a dysfunction of LSD1 activity.
In an additional aspect, LSD activity dysfunction is a dysfunction of LSD2 activity. In yet another aspect, the invention relates to the use of the disclosed compound in the manufacture of a medicament for the treatment of an uncontrolled cell proliferation disorder.
Also disclosed are the uses of a disclosed compound or a disclosed product in the manufacture of a medicament for the treatment of a disorder associated with an LSD dysfunction in a mammal.
Although aspects of the present invention may be described and claimed in a particular legal class, such as the legal system class, this is done for convenience only, and one 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, it is by no means intended that any method or aspect defined in this document be construed as requiring that its steps be performed in a specific order. In that sense, where a method claim does not specifically define in the claims or descriptions that the steps must be limited to a specific order, it is by no means intended to be inferred in any respect. This applies to any possible unstated basis for interpretation, including questions of logic regarding the arrangement of steps or operational flow, the simple meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the descriptive report.
DESCRIPTION
The present invention may be more readily understood by reference to the following detailed description of the invention and the examples included therein.
Before the present compounds, compositions, articles, systems, devices and / or methods are disclosed and described, it should be understood that they are not limited to specific synthetic methods unless otherwise specified or to reagents. particular, unless otherwise specified, as they may of course vary. It should also be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting. While any methods and materials similar or equivalent to those described in the present invention may be used in the practice or testing of the present invention, exemplary methods and materials will now be described.
All publications mentioned in this document are hereby incorporated by reference to disclose and describe the methods and / or materials for which the publications are cited. The publications discussed in this document are provided solely for disclosure prior to the filing date of this application. Nothing herein should be construed as an admission that the present invention is not intended to precede such publication by virtue of the foregoing invention. In addition, the publication dates stated in this document may differ from the actual publication dates, which may require independent confirmation.
A. DEFINITIONS
As used in the present invention, nomenclature for compounds, including organic compounds, may be provided using common names or recommendations from IUPAC, IUBMB or CAS for nomenclature.
When one or more stereochemical characteristics are present, the Cahn-Ingold-Prelog rules for stereochemistry may be employed to designate stereochemical priority, the E / Z specification and the like. One of ordinary skill in the art can easily determine the structure of a compound if a name is given by systemically reducing the structure of the compound using naming conventions, or by commercially available software such as ChemDraw ™ (Cambridgesoft Corporation, U.S.A.).
As used in the specification and the appended claims, the singular forms "one", "one" and "the" include plural references, unless the context clearly states otherwise. Thus, for example, reference to "a functional group," "an alkyl" or "a residue" includes mixtures of two or more such functional groups, alkyls or residues, and the like.
The ranges may be expressed herein as from "about" a particular value, and / or even "about" another particular value. When such a range is expressed, another aspect includes from a particular value and / or even another particular value. Similarly, when values are expressed as approximations, by using the predecessor "about", it will be understood that the particular value forms another aspect. It will further be understood that the parameters of each of the ranges are significant both with respect to the other parameter and independently of the other parameter. It is also understood that there are various values disclosed in this document, and that each value is also disclosed in this document 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 specific units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13 and 14 are also disclosed.
References in the specification and conclusive claims for parts by weight of a particular element or component in a composition denotes the weight ratio between the element or component and any other elements or components in the composition or article for which a part by weight. is expressed. 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 weight ratio of 2: 5, and are present in such ratio regardless of whether the additional components are present. contained in the compound.
A percentage by weight (% by weight) of a component, unless specifically stated otherwise, is based on the total weight of the formulation or composition in which the component is included.
As used in the present invention, the term "LSD" collectively refers to one or both LSD1 and LSD2.
As used in the present invention, the terms "LSD1" and "specific lysine demethylase 1" may be used interchangeably and refer to a histone demethylase encoded by the KDM1A gene. The KDM1A gene has a locus in the lp36.12 gene map, as described by the cytogenetic band Gene Entrez, the cytogenetic band Ensembl, and the cytogenetic band HGNC. The term LSD1 refers to a native protein that has 852 amino acids with molecular weight of about 92903 Da, and is a member of the flavamine monoamine oxidase family. The term LSD1 is inclusive of protein, gene product and / or gene referred to by such alternative names as: LSD1, KDM1; RP1-184J9.1; AOF2; BHCl110; KIAA0601; LSD1; BRAF35-HDAC BHC110 complex protein; BRAF35-HDAC FAD binding protein complex, 110 KDa subunit; amine oxidase 2 domain (containing flavin); histone demethylase 1 specific lysine; histone demethylase IA; flavin-containing amine oxidase domain protein 2; lysine specific demethylase 1 (K); amine oxidase domain 2 (containing flavin); and 110 Kda subunit of the BRAF35-HDAC FAD-binding protein complex as used by those skilled in the art.
As used in the present invention, the terms "LSD2" and "specific lysine demethylase 2" may be used interchangeably and refer to a histone demethylase encoded by the KDM1B gene. The KDM1B gene has a gene map locus of 6p22.3, as described by the cytogenetic band Gene Entrez, the cytogenetic band Ensembl, and the cytogenetic band HGNC. The term LSD21 refers to a native protein that has 822 amino acids with molecular weight of about 92098 Da, and is a member of the flavamine monoamine oxidase family. The term LSD2 is inclusive of protein, gene product and / or gene referred to by such alternative names as: LSD2, AOF1; FLJ33898; FLJ34109; FLJ43328; C6orfl93; DKFZp686I0412; OTTHUMP00000179125; bA204B7.3; dJ298J15.2; flavin-containing amine oxidase domain protein 1; lysine-2 specific histone demethylase; specific lysine (K) demethylase 1B; amine oxidase domain 1 (containing flavin); flavine 1-containing amine oxidase; lysine-2 specific histone demethylase; open reading frame 193 of chromosome 6; and specific lysine histone demethylase 1B; as used by those skilled in the art.
As used herein, the term "histone demethylase" refers to that group of enzymes that remove methyl groups from histone proteins. The term is inclusive of lysine histone demethylase, ie enzymes that remove methyl groups from lysine residues in histones, and arginine histone demethylase, ie enzymes that remove methyl groups from arginine residues in histones.
As used in the present invention, the term "histone lysine demethylase" or "specific lysine histone demethylase" may be used interchangeably, and both refer to the group of enzymes that remove methyl groups from lysine residues of histone proteins. . Lysine histone demethylases are a group of enzymes comprising the following specific forms: LSD1, LSD2, JMJD2A, JMJD2B, JMJD2C and JMJD2D.
As used herein, "optional" or "optionally" means that the subsequently described event of circumstances may or may not occur, and that the description includes examples where said event or circumstance occurs, and examples where it does not occur. .
As used in the present invention, the term "individual" may be a vertebrate, such as a mammal, a fish, a bird, a reptile or an amphibian. Thus, the subject of the methods disclosed herein may be a human being, a nonhuman primate, a 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 newborn individuals, such as fetuses, male and female, are intended to be encompassed. In one aspect, the individual is a mammal. A patient refers to an individual afflicted with a disease or disorder. The term "patient" includes both human and veterinary individuals. In some aspects of the disclosed methods, the subject has been diagnosed with a need for treatment of an uncontrolled cell proliferation disorder associated with a histone lysine demethylase dysfunction prior to the administration step. In some aspects of the disclosed method, the subject was diagnosed with a need for inhibition of a histone lysine demethylase prior to the administration step.
As used in the present invention, the term "treatment" refers to the medical management of a patient with the intention of curing, ameliorating, stabilizing or preventing a disease, condition or disorder. This term includes active treatment, ie treatment directed specifically at amelioration of a disease, pathological condition or disorder, and also includes causal treatment, ie treatment aimed at removing the cause of the disease, pathological condition or of the associated disorder. In addition, this term includes palliative treatment, ie treatment designed to relieve symptoms rather than cure the disease, condition, or disorder; preventive treatment, ie treatment directed to minimize or partially or completely inhibit the development of the associated disease, pathological condition or disorder; and supportive treatment, that is, treatment employed to supplement other specific therapy directed at ameliorating the associated disease, pathological condition, or disorder. In many respects, the term encompasses any treatment of an individual, including a mammal (for example, a human) and includes: (i) preventing disease from occurring in an individual who may be predisposed to the disease but has not yet been diagnosed as having; (ii) inhibit the disease, i.e. stop its development; or (iii) alleviate the disease, ie, cause regression of the disease. In one aspect the individual is a mammal like a primate and in another aspect the individual is a human being. The term "individual" also includes domesticated animals (e.g., cats, dogs, etc.), cattle (e.g., cattle, horses, pigs, sheep, goats, etc.) and laboratory animals (e.g., mouse, rabbit). , rat, guinea pig, fruit fly, zebrafish, etc.).
As used in the present invention, the term "preventing" or "preventing" refers to preventing, preventing, preventing, anticipating, interrupting or hindering something from happening, especially by anticipatory action. It is understood that where reducing, inhibiting or preventing is used herein, unless specifically indicated otherwise, the use of the other two words is also expressly disclosed.
As used in the present invention, the term "diagnosed" means having undergone a physical examination by a person skilled in the art, for example a physician, who has been found to have a condition that can be diagnosed or treated by the compounds. , compositions or methods disclosed herein. For example, "diagnosed with an uncontrolled cell proliferation disorder" means having undergone a physical examination by a person skilled in the art, such as a physician, and finding a condition that can be diagnosed or treated by a compound or composition. which may inhibit a histone lysine demethylase. As an additional example, "diagnosed with a need for inhibition of a histone demethylase" refers to having undergone a physical examination by a person skilled in the art, for example a physician, and discovering a condition characterized by a dysfunction. of histone demethylase. Such a diagnosis may be in reference to a disorder, such as an uncontrolled cell proliferation disorder, cancer, and the like, as discussed in this document. For example, the term "diagnosed with a need for inhibition of histone demethylase activity" refers to having undergone a physical examination by a person skilled in the art, for example a physician, and discovering a condition that may be diagnosed or treated by inhibition of histone demethylase activity. For example, "diagnosed with a need to treat one or more uncontrolled cell proliferation disorders associated with a histone demethylase dysfunction" means having undergone a physical examination by a person skilled in the art, such as a physician, and discovering have one or more uncontrolled cell proliferation disorders associated with a histone demethylase dysfunction.
As used in the present invention, the phrase "identified as in need of treatment for a disorder" or the like, refers to the selection of an individual as a basis for the need for treatment of the disorder. For example, an individual may be identified as having a need for the treatment of a disorder (e.g., a disorder related to a histone demethylase activity disorder) based on an earlier diagnosis by a person skilled in the art and thereafter. undergo treatment for the disorder. It is contemplated that identification may, in one aspect, be performed by a person other than the person making the diagnosis. It is also contemplated, in another aspect, that the administration may be performed by a person who subsequently performed the administration.
As used herein, the terms "administering" and "administering" refer to any method of providing a pharmaceutical preparation to an individual. 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, ophthalmic administration, intraaural administration, intracerebral administration, rectal, 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, a preparation may be administered therapeutically; that is, administered to treat an existing disease or condition. In other various aspects, a preparation may be administered prophylactically; ie administered to prevent a disease or condition.
The term "contacting" as used herein refers to bringing together a disclosed compound and a cell, target receptor or other biological entity such that the compound may affect the activity of the target (e.g. receiver, cell, etc.), directly; that is, by interacting with the target itself, or indirectly; that is, by interacting with another molecule, cofactor, factor, or protein on which target activity is dependent.
As used in the present invention, the terms "effective amount" and "effective amount" refer to an amount that is sufficient to achieve the desired result or to have an effect on an unwanted condition. For example, a "therapeutically effective amount" refers to an amount that is sufficient to achieve the desired therapeutic outcome or to have an effect on undesirable symptoms, but is not generally sufficient to cause adverse side effects. The specific therapeutically effective dose level for any particular patient will depend on a number of factors, including the disorder being treated and the severity of the disorder; the specific composition used; the age, body weight, general health, gender and diet of the patient; the time of administration, the route of administration; the excretion rate of the specific compound used; the duration of treatment; medicines used in combination or at the same time with the specific compound employed, and similar factors well known in medical techniques. For example, it is well within the ability of the person skilled in the art to initiate doses of a compound at levels below that necessary to achieve the desired therapeutic effect and to gradually increase the dose to the desired effect. If desired, the effective daily dose may be divided into several doses for administration purposes. Accordingly, single dose compositions may contain such amounts or submultiples thereof to make up the daily dose. The dosage may be adjusted by the individual's physician in case there are any contraindications. The dosage may vary and may be administered in one or more daily dose administrations for one or several days. Guidance for suitable dosages for certain classes of pharmaceutical products can be found in the literature. In various additional aspects, a preparation may be administered in a "prophylactically effective amount"; that is, an amount effective to prevent a disease or condition.
As used herein, "EC50" refers to the concentration of a substance (e.g., a compound or a drug) that is required to achieve 50% agonism or activation of a biological process, or component of a including a protein, subunit, organelle, ribonucleoprotein, etc. In one aspect, an EC50 may refer to the concentration of a substance that is required for 50% agonism or activation in vivo, as further defined elsewhere herein. In another aspect, the EC50 refers to the concentration of agonist or activator that elicits a midway response between the baseline and the maximal response.
As used herein, "IC50" refers to the concentration of a substance (e.g., a compound or a drug) that is required to achieve 50% inhibition of a biological process, or component of a process, including a protein, subunit, organelle, ribonucleoprotein, etc. For example, an IC 50 may refer to the concentration of a substance that is required for 50% inhibition in vivo or the inhibition is measured in vitro, as further defined elsewhere herein. Alternatively, the IC50 refers to half of the maximum inhibitory concentration (IC) (50%) of a substance. Inhibition can be measured in a cell line 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. In yet a further aspect, inhibition is measured in a cell line, e.g. HEK-293 or HeLa, transfected with a wild type or mutant mammalian histone demethylase, for example LSD1 or LSD2.
The term "pharmaceutically acceptable" describes a material that is not biologically or otherwise undesirable, that is, that does not cause an unacceptable level of undesirable or deleteriously interacting biological effects.
The term "stable" as used herein refers to compounds which are not substantially altered when subjected to conditions permitting their production, detection and, in certain respects, their recovery, purification and use for a or more of the purposes disclosed herein.
As used herein, the term "derivative" refers to a compound that has a structure derived from the structure of an parent compound (e.g., a compound disclosed in the present invention) and whose structure is sufficiently similar to those disclosed. in that document and, based on that similarity, a person skilled in the art would expect identical or similar activities and uses to the claimed compounds, or to induce, as a precursor, identical or similar activities and uses to the claimed compounds. Exemplary derivatives include salts, esters, amides, ester or amide salts and N-oxides of a parent compound.
As used herein, the term "pharmaceutically acceptable carrier" 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 just prior to use. Examples of suitable aqueous or non-aqueous vehicles, 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. as ethyl oleate. Proper flowability can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in case of dispersions and by the use of surfactants. These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. The prevention of the action of microorganisms can be ensured by the inclusion of several antibacterial and antifungal agents, such as paraben, chlorobutanol, phenol, sorbic acid and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form may be caused by the inclusion of agents such as aluminum monostearate and gelatin which delay absorption. Injectable depot forms are made by forming microencapsulation matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, (poly) orthoesters and (poly) anhydrides. Depending on the drug to polymer ratio and the nature of the particular polymer employed, the rate of drug release may be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues. Injectable formulations may be sterilized, for example, by filtration through a bacterial retention filter, or by incorporation of sterilizing agents in the form of sterile solid compositions which may be dissolved or dispersed in sterile water or other sterile injectable media 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 micrometers.
A residue of a chemical species, as used in the specification and the final claims, refers to the portion which is the resulting product of the chemical species in a particular reaction scheme, subsequent formulation or chemical, regardless of whether the portion It is effectively obtained from chemical species. Thus, an ethylene glycol residue in a polyester refers to one or more -OCH 2 CH 2 O- units in the polyester, regardless of whether ethylene glycol is used to prepare the polyester. Similarly, a sebacic acid residue in a polyester refers to one or more -CO (CH2) sCO- portions in the polyester, regardless of whether the residue is obtained by reacting sebacic acid or an ester thereof to obtain the polyester.
As used in the present invention, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the allowable 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. The allowable substituents may be one or more and the same or different for appropriate organic compounds. For disclosure purposes, heteroatoms, such as nitrogen, may have hydrogen substituents and / or any allowable substituents of the organic compounds described herein that satisfy the valencies of the heteroatoms. This disclosure is not intended to be limited in any way by the allowable substituents of the organic compounds. In addition, the terms "substitution" or "substituted for" include the implicit caveat that such substitution is in accordance with the permitted valence of the substituted atom and substituent, and that substitution results in a stable compound, for example, in a compound that does not undergo spontaneous transformation such as rearrangement, cyclization, elimination, etc. It is also contemplated that in certain aspects, unless expressly indicated otherwise, individual substituents may be further optionally substituted (i.e. further substituted or unsubstituted).
In the definition of various terms, "A1", "A2", "A3" and "A4" are used herein as generic symbols to represent various specific substituents. These symbols may be any substituent, not limited to those disclosed herein, and when they are defined to be certain substituents in one example, they may, in another example, be defined as some other substituents.
The term "alkyl" as used herein is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-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. The alkyl group may also be substituted or unsubstituted. For example, the alkyl group may be substituted with one or more groups including, but not limited to alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxyl, nitro, silyl, sulfo-oxo or thiol as described herein invention. A "lower alkyl" group is an alkyl group containing from one to six years (for example from one to four) carbon atoms.
For example, a "C1-C3 alkyl" group may be selected from methyl, ethyl, n-propyl, i-propyl and cyclopropyl, or a subgroup thereof. In certain aspects, the "C1-C3 alkyl" group may optionally be further substituted. As a further example, a "C1-C4 alkyl" group may be selected from methyl, ethyl, n-propyl, i-propyl, cyclopropyl, n-butyl, i-butyl, s-butyl, t-butyl and chlorobutyl, or of a subset of them. In certain aspects, the "C1-C4 alkyl" group may optionally be further substituted. As a further example, a "C1-C6 alkyl" group may be selected from methyl, ethyl, n-propyl, i-propyl, cyclopropyl, n-butyl, i-butyl, s-butyl, t-butyl, chloroformate, n. -pentyl, i-pentyl, s-pentyl, t-pentyl, neopentyl, cyclopentyl, n-hexyl, i-hexyl, 3-methylpentane, 2,3-dimethylbutane, neohexane and cyclohexane, or a subset thereof. In certain aspects, the "C1-C6 alkyl" group may optionally be further substituted. As a further example, a "C1 -C6 alkyl" group may be selected from methyl, ethyl, n-propyl, i-propyl, cyclopropyl, n-butyl, i-butyl, s-butyl, t-butyl, chlorobutyl, -pentyl, i-pentyl, s-pentyl, t-pentyl, neopentyl, cyclopentyl, n-hexyl, i-hexyl, 3-methylpentane, 2,3-dimethylbutane, neohexane, cyclohexane, heptane, cycloheptane , octane and cyclooctane, or a subset thereof. In certain aspects, the "C1 -C6 alkyl" group may optionally be further substituted. As a further example, a "C1-C12 alkyl" group may be selected from methyl, ethyl, n-propyl, i-propyl, cyclopropyl, n-butyl, i-butyl, s-butyl, t-butyl, chlorobutyl, -pentyl, i-pentyl, s-pentyl, t-pentyl, neopentyl, cyclopentyl, n-hexyl, i-hexyl, 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, the "C1-C12 alkyl" group may optionally be further substituted.
Throughout the specification, "alkyl" is generally used to refer to unsubstituted alkyl groups and substituted alkyl groups; however, substituted alkyl groups are also specifically referred to herein by identifying the specific substituent (s) on the alkyl group. For example, the term "halogenated alkyl" or "haloalkyl" refers specifically to an alkyl group that is substituted with one or more halides, for example fluorine, chlorine, bromine or iodine. The term "alkoxyalkyl" refers specifically to an alkyl group that is substituted with one or more alkoxy groups as described below. The term "alkylamino" refers specifically to an alkyl group that is substituted with one or more amino groups as described below and the like. When "alkyl" is used in one example and a specific term such as "alkyl alcohol" is used in another, this does not imply that the term "alkyl" also does not refer to specific terms such as "alkyl alcohol" and the like. .
This practice is also used for other groups described in this document. That is, while a term such as "cycloalkyl" refers to both substituted and unsubstituted cycloalkyl moieties, the substituted moieties may furthermore be specifically identified herein; for example, a particular substituted cycloalkyl may be referred to as, for example, an "alkylcycloalkyl". Similarly, a substituted alkoxy may be specifically referred to as, for example, a "halogenated alkoxy", a particular substituted alkenyl may be, for example, an "alkenyl alcohol" and the like. Again, the practice of using a general term such as "cycloalkyl" and a specific term such as "alkylcycloalkyl" should not imply that the general term also does not include the specific term.
The term "cycloalkyl" as used in the present invention is a non-aromatic carbon-based ring composed 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 type of cycloalkyl group as defined above and is included within the meaning of the term "cycloalkyl" where at least one of the ring carbon atoms is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus. The cycloalkyl group and the heterocycloalkyl group may be substituted or unsubstituted. The cycloalkyl group and the heterocycloalkyl group may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxyl, nitro, silyl, sulfo-oxo, nitrile, sulfonamide or thiol as described in the present invention.
The term "polyalkylene group" as used in the present invention is a group with two or more CH2 groups attached to each other. The polyalkylene group may be represented by the formula - (CH2) a-, where "a" is an integer from 2 to 500.
The terms "alkoxy" and "alkoxy" as used in the present invention refer to an alkyl or cycloalkyl group bonded via an ether bond; that is, an "alkoxy" group may be defined as -OA1, where A1 is alkyl or cycloalkyl as defined above. "Alkoxy" also includes polymers of alkoxy groups as just described; that is, an alkoxy may be a polyether such as -OA1-OA2 or -OA1- (OA2) a-OA3, where "a" is an integer from 1 to 200 and A1, A2 and A3 are alkyl and / or cycloalkyl.
The term "alkenyl" as used herein is a hydrocarbon group of 2 to 24 carbon atoms with a structural formula containing at least one carbon-carbon double bond. Asymmetric structures, such as (A ^ -A2) C = C (A3A4), include both E and Z isomers. This can be assumed from the structural formulas herein, where an asymmetric alkene is present, or can be explicitly indicated by the symbol. Binding C = C. The alkenyl group may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfo-oxo, nitrile, sulfonamide or thiol as described in the present invention.
The term "cycloalkenyl" as used herein is a non-aromatic carbon-based ring composed of at least three carbon atoms and containing at least one carbon-carbon double bond, that is, C = C. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, norbornenyl and the like. The term "heterocycloalkenyl" is a type of cycloalkenyl group as defined above and is included within the meaning of the term "cycloalkenyl" where at least one of the ring carbon atoms is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus. The cycloalkenyl group and the heterocycloalkenyl group may be substituted or unsubstituted. The cycloalkenyl group and the heterocycloalkenyl group may 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, halide, hydroxyl, ketone, azide, nitro, silyl, sulfo-oxo, nitrile, sulfonamide or thiol as described in the present invention.
The term "alkynyl" as used herein is a hydrocarbon group of 2 to 24 carbon atoms with a structural formula containing at least one carbon-carbon triple bond. The alkynyl group may be unsubstituted or substituted by one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkylyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether , halide, hydroxyl, ketone, azide, nitro, silyl, sulfo-oxo, nitrile, sulfonamide or thiol as described in the present invention.
The term "cycloalkynyl" as used herein is a non-aromatic carbon-based ring composed of at least seven carbon atoms and containing at least one carbon-carbon triple bond. Examples of cycloalkynyl groups include, but are not limited to, cycloheptinyl, cyclooctinyl, cyclononinyl and the like. The term "heterocycloalkynyl" is a type of cycloalkenyl group as defined above and is included within the meaning of the term "cycloalkynyl" where at least one of the ring carbon atoms is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus. The cycloalkyl group and the heterocycloalkyl group may be substituted or unsubstituted. The cycloalkyl group and the heterocycloalkyl group may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfo-oxo, nitrile, sulfonamide or thiol as described in the present invention.
The term "aryl" as used in the present invention 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 that has 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. Similarly, the term "non-heteroaryl", which is also included in the term "aryl", defines a group that contains an aromatic group that does not contain a heteroatom. The aryl group may be substituted or unsubstituted. The aryl group may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfo-oxo, nitrile, sulfonamide or thiol as described in the present invention. The term "biaryl" is a specific type of aryl group and is included in the definition of "aryl". Biaryl refers to two aryl groups which are attached via a fused ring structure, as in naphthalene, or which are linked through one or more carbon-carbon bonds, as in biphenyl.
The term "aldehyde" as used in the present invention is represented by the formula -C (O) H. Throughout this descriptive report, "C (O)" is an abbreviated notation for a carbonyl group, ie, C = O.
The terms "amine" or "amino" as used in the present invention are represented by the formula -NA1 -A2, where A1 and A2 may independently be hydrogen or alkyl, cycloalkyl, alkenyl, cycloalkenyl group , alkynyl, cycloalkylyl, aryl or heteroaryl as described in the present invention.
The term "alkylamino" as used in the present invention is represented by the formula -NH (-alkyl), where alkyl is as described herein. Representative examples include, but are not limited to, methylamino group, ethylamino group, propylamino group, isopropylamino group, butylamino group, isobutylamino group, (sec-butyl) amino group, (tert-butyl) amino group, pentylamino group, isopentylamino group, (tert-pentyl) amino group, hexylamino group and the like.
The term "dialkylamino" as used in the present invention is represented by the formula -N (-alkyl) 2, where alkyl is as described herein. Representative examples include, but are not limited to, dimethylamino group, diethylamino group, dipropylamino group, diisopropylamino group, dibutylamino group, diisobutylamino group, di (sec-butyl) amino group, di (tert-butyl) amino group, dipentylamino group, diisopentylamino group, di (tert-pentyl) amino group, dihexylamino group, N-ethyl-N-methylamino group, N-methyl-N-propylamino group, N-ethyl-N-propylamino group and the like .
The term "carboxylic acid" as used in the present invention is represented by the formula -C (O) OH.
The terms "ester" as used in the present invention is represented by the formula -OCOJA1 or -CIOJOA1, where A1 may be alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkyl, aryl or heteroaryl as described herein . The term "polyester" as used in the present invention is represented by the formula - (A 2 O (O) C-A2-C (O) 0) a - or - (A 2 O 1 O 2 -C 2 -OC (O)) a-, where A1 and A2 may be independently an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group described herein and "a" is an integer from 1 to 500. "Polyester" is The term is used to describe a group that is produced by the reaction between a compound with at least two carboxylic acid groups with a compound with at least two hydroxyl groups.
The term "ether" as used in the present invention is represented by the formula A 1 -OA 2, wherein A 1 and A 2 may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl as described in the present invention. The term "polyether" as used in the present invention is represented by the formula - (A 2 O-A 2 O) a -, where A 1 and A 2 may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl described herein and "a" is an integer from 1 to 500. Examples of polyether groups include polyethylene oxide, polypropylene oxide and polybutylene oxide.
The terms "halogen," "halide" and "halo" as used herein refer to the halogens fluorine, chlorine, bromine and iodine. It is also envisaged that in various respects halogen may be selected from fluorine, chlorine, bromine and iodine. For example, halogen may be selected from fluorine, chlorine and bromine. As another example, halogen may be selected from fluorine and chlorine. As another example, halogen may be selected from chlorine and bromine. As another example, halogen may be selected from bromine and iodine. As another example, halogen may be selected from chlorine, bromine and iodine. In one aspect, halogen may be fluorine. In another aspect, halogen may be chlorine. In another additional aspect, halogen is bromine. In an additional aspect, halogen is iodine.
It is also envisaged that in certain respects pseudohalogens (eg, triflate, mesylate, tosylate, brosylate, etc.) may be used instead of halogens. For example, in certain respects halogen may be substituted for a pseudohalogen. As a further example, pseudohalogen may be selected from triflate, mesylate, tosylate and brosylate. In one aspect, the pseudohalogen is triflate. In another aspect, the pseudohalogen is mesylate. In another aspect, pseudohalogen is tosylate. In another aspect, pseudohalogen is brosylate.
The term "heterocycle" as used herein refers to mono- or multicyclic aromatic or non-aromatic ring systems, wherein at least one ring member is different from carbon.
Heterocycle includes 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, pyrrol, pyrrolidine, tetrahydrofuran, tetrahydropyran, tetrazine, including 1,2,4-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-triazol, 1,3,4-triazol and the like.
The term "hydroxyl" as used in the present invention is represented by the formula -OH.
The term "ketone" as used in the present invention is represented by the formula wherein R1 and R2 may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl as described in the present invention.
The term "azide" as used in the present invention is represented by the formula -N3.
The term "nitro" as used in the present invention is represented by the formula -NO 2.
The term "nitrile" as used in the present invention is represented by the formula -CN.
The term "silyl" as used in the present invention is represented by the formula -SiA1A2A3, wherein A1, A2 and A3 may independently be hydrogen or an alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl aryl or heteroaryl as described in the present invention.
The term "sulfo-oxo" as used in the present invention is represented by the formulas -SYJA1, -S (O) 2A1, -OS (O) 2A1 or -OS (O) 2OA1, where A1 may be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described in the present invention. Throughout this descriptive report, "S (O)" is an abbreviated notation for S = O. The term "sulfonyl" is used herein to refer to the sulfo-oxo group represented by the formula —S (O) 2A1 'where A1 may be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl as described in the present invention. The term "sulfone" as used in the present invention is represented by the formula A1S (O) 2A2, where A1 and A2 may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described in the present invention. The term "sulfoxide" as used in the present invention is represented by the formula wherein Al and A2 may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein. in the present invention.
The term "thiol" as used in the present invention is represented by the formula -SH.
"R1", "R2", "R3" and "Rn" where n is an integer as used in the present invention may independently have one or more of the groups listed above. For example, if R1 is 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 and the like. Depending on the selected groups, a first group may be incorporated within the second group, or alternatively, the first group may be hung (ie attached) to the second group. For example, with the phrase "an alkyl group comprising an amino group", the amino group may be incorporated into the alkyl group structure. Alternatively, the amino group may be attached to the alkyl group structure. The nature of the group (s) that is / will be selected will determine if the first group is incorporated or attached to the second group.
As described in the present invention, the compounds of the invention may contain "optionally substituted" moieties. In general, the term "substituted", whether preceded by the term "optionally" or not, means that one or more hydrogens of the designated moiety are substituted with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position in the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group. , the substituent may be the same or different at each position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically viable compounds. It is also contemplated that in certain aspects, unless expressly indicated otherwise, individual substituents may be further optionally substituted (i.e. further substituted or unsubstituted).
Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen; - (CH 2) o -4R °; - (CH 2) o -40 R °; -O (CH 2) 0-4 R0, -O- (CH 2) 0-4C (O) OR °; - (CH 2) 0-4CH (OR °) 2; - (CH2) 0-4SR0; - (CH2) o-4Ph, which may be substituted with R °; - (CH 2) 0-4 O (CH 2) o-iPh, which may be substituted with R °; - CH = CHPh, which may be substituted with R °; - (CH 2) 0-4 O (CH 2) 0-1 -pyridyl, which may be substituted with R °; -NO2; -CN; - N3; - (CH 2) 0-4 N (R °) 2; - (CH 2) 0-4 N (R °) C (O) R °; -N (R °) C (S) R °; - (CH 2) o -4N (R °) C (O) NR ° 2; -N (R °) C (S) NR ° 2; - (CH 2) 0-4 N (R °) C (O) 0 R °; N (R °) N (R °) C (0) R °; -N (R °) N (R °) C (O) NR ° 2; -N (R °) N (R °) C (O) 0 R °; - (CH 2) 0-4C (0) R °; -C (S) R °; - (CH 2) 0-4C (0) 0R °; - (CH 2) o -4C (O) SR °; - (CH 2) 0-4 ° C (0) OSi R ° 3; - (CH 2) 0-4 ° C (0) R °; -0C (0) (CH2) 0-4SR-, SC (S) SR °; - (CH 2) 0-4 ° C (0) R °; - (CH 2) 0-4C (0) NR ° 2; -C (S) NR ° 2; -C (S) SR °; -SC (S) SR °, - (CH 2) 0-4OC (0) NR ° 2; -C (O) N (OR °) R °; C (O) C (O) R °; -C (O) CH 2 C (O) R °; -C (NOR °) R °; - (CH 2) 0-4SSR0; - (CH 2) o -4S (O) 2 R °; - (CH 2) 0-4 S (O) 20 R °; - (CH 2) o -40 S (O) 2 R °; S (O) 2 NR ° 2; - (CH 2) 0-4 S (0) R °; -N (R °) S (O) 2 NR ° 2; -N (R °) S (O) 2 R °; - N (0R °) R °; -C (NH) NR ° 2; -P (O) 2 R °; -P (O) R ° 2; -OP (O) R ° 2; 0P (0) (0R °) 2; SiR ° 3; - (straight or branched chain C1-4 alkylene) 0-N (R °) 2; or - (C1-4 straight or branched chain alkylene) C (0) 0-N (R °) 2, wherein each R ° may be substituted as defined below and is independently hydrogen, a C1-6 aliphatic, -CH2Ph , -0 (CH2) o-iPh, or a saturated, partially unsaturated ring or 5 to 6 membered aryl of 0 to 4 heteroatoms selected independently of nitrogen, oxygen or sulfur or, despite the above definition, two independent occurrences of R °, together with their intermediate atom (s), form a 3- to 12-membered saturated, partially unsaturated or bicyclic mono- or bicyclic ring with 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.
Suitable monovalent substituents on R ° (or on the ring formed by taking two independent occurrences of R ° together with its intermediate atoms), are independently halogen, - (CH 2) o-2 R ·, - (halo R ·), - (CH 2 ) o-2OH, - (CH2) 0-2OR ·, - (CH2) o-2CH (OR ') 2; -0 (halo R ·), -CN, -N 3, - (CH 2) o-2C (0) R ·, - (CH 2) o-2C (0) OH, - (CH 2) o-2C (0) OR · , - (CH2) o-2SR ·, - (CH2) o-2SH, - (CH2) 0-2NH2, - (CH2) o-2NHR ·, - (CH2) o-2NR '2, -no2, - SiR ®3, -OSíR®3, -C (O) SR ·, - (linear or branched C1-4 alkylene) C (O) OR · or -SSR ·, where each R · is unsubstituted, or where preceded by "halogen" is substituted with only one or more halogens, and is independently selected from C1-4 aliphatics, -CH2Ph, -0 (CH2) o-iPh, or a saturated, partially unsaturated ring or 5 to 6 membered aryl, with 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents on a saturated carbon atom of R ° include = O and = S.
Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: = 0, = S, = NNR * 2, = NNHC (O) R *, = NNHC (O) OR *, = NNHS (O) 2R *, = NR *, = NOR *, -O (C (R * 2)) 2-3O-, or -S (C (R * 2)) 2-3S-, where each Independent occurrence of R * is selected from hydrogen, a C1-6 aliphatic which may be substituted as defined below, or a saturated, partially unsaturated ring or 5 to 6 membered aryl and with 0 to 4 heteroatoms selected independently of nitrogen, oxygen or sulfur. Suitable divalent substituents that bind to the vicinal substitutable carbons of an "optionally substituted" group include: -O (CR * 2) 2-3O-, wherein each independent occurrence of R * is selected from hydrogen, a C1-6 aliphatic which may be substituted as defined below or a saturated, partially unsaturated ring or 5-6 membered aryl having 0 to 4 heteroatoms selected independently from nitrogen, oxygen or sulfur.
Suitable substituents on the aliphatic group R * include halogen, -R ·, - (halo R ·), -OH, -OR ·, -0 (halo R ·), -CN, -C (O) OH, -C ( O) OR ·, -NH2, -NHR ·, -NR »2, or -NO2, wherein each R · is unsubstituted or, where preceded by" halo ", is substituted with only one or more halogens and is independent of one. C1-4 aliphatic, -0 (CH2) o-iPh or a saturated, partially unsaturated ring or 5-6 membered aryl with 0 to 4 heteroatoms selected independently of nitrogen, oxygen or sulfur.
Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R +, -NR + 2, -C (O) R +, -C (O) OR +, -C (O) C (O) R +, -C (O) CH 2 C (O) R +, -S (O) 2 R +, -S (O) 2 NR + 2, -C (S) NR + 2, -C (NH) NR + 2 or -N (R +) S (O) 2 R +; wherein each Rt is independently hydrogen, a C1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh or a saturated, partially unsaturated or unsubstituted 5 to 6 membered aryl ring of 0 to 4 heteroatoms selected independently of notwithstanding the above definition, two independent occurrences of Rf, taken together with its intermediate atom (s) form a partially unsaturated or partially saturated or bicyclic ring 3 to 12 membered aryl, with 0 to 4 heteroatoms selected independently of nitrogen, oxygen or sulfur.
Suitable substituents on the aliphatic group R + are independently halogen, -R ·, - (halo R ·), -OH, -OR ·, -O (halo R ·), -CN, -C (O) OH, -C ( O) OR ·, -NH2, -NHR ·, -NR®2, or -NO2, where each R · is unsubstituted or, where preceded by 'halo', is substituted with only one or more halogens and is independent of one. C 1-4 aliphatic, -CH 2 Ph, -0 (CH 2) o-iPh or a saturated, partially unsaturated ring or 5-6 membered aryl with 0 to 4 heteroatoms selected independently of nitrogen, oxygen or sulfur.
The term "leaving group" refers to an electron-removing atom (or group of atoms) that can be displaced as a stable species, carrying the electrons of the bond with it. Examples of suitable leaving groups are halides - including chlorine, bromine and iodine - and pseudohalides (sulfonate esters) - including triflate, mesylate, tosylate and brosylate. It is also contemplated that a hydroxyl moiety may be converted to a leaving group by a Mitsunobu reaction.
The terms "hydrolysable group" and "hydrolysable moiety" refer to a functional group capable of hydrolysis, for example under acidic or basic conditions. Examples of hydrolysable residues include, without limitation, acid halides, activated carboxylic acids and various protecting groups known in the art (see, for example, Protective Groups in Organic Synthesis, T.W. Greene, P.G. Wuts, Wiley-Interscience, 1999).
The term "protecting group" means a group that protects one or more functional groups of a compound, giving rise to a protected derivative of the specified compound. Functional groups which may be protected include, by way of example, amino groups, hydroxyl groups and the like. Protecting groups are well known to those skilled in the art and are described, for example, in T.W. Greene and G.M. Wuts, Protective Groups in Organic Synthesis, Third Edition, Wiley, New York, 1999, and references cited therein.
The term "amino protecting group" means a protecting group suitable for preventing unwanted reactions in an amino group, and includes, but is not limited to tert-butoxycarbonyl (BOC), trityl (Tr), benzyloxycarbonyl (Cbz), 9-fluorenylmethoxycarbonyl (FMOC), formyl, trimethylsilyl (TMS), tert-butyldimethylsilyl (TBS), benzyl, p-methoxybenzyl, p-fluorobenzyl, p-chlorobenzyl, diphenylmethyl naphthylmethyl, tetrahydropyran and THP).
The term "hydroxyl protecting group" means a suitable protecting group to prevent undesirable reactions in a hydroxyl group. Representative hydroxyl protecting groups include, but are not limited to, silyl groups including trialkylsilyl (C1-6) groups such as trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBS) and the like; esters (acyl groups) including (C 1-6) alkanoyl groups such as formyl, acetyl and the like; arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), diphenylmethyl (benzhydryl, DPM), tetrahydropyran (THP), methoxymethyl (MOM), methylthiomethyl (MTM), benzyloxymethyl (BOM) ) and the like.
The term "organic residue" defines a carbon-containing residue, that is, a residue comprising at least one carbon atom and includes, but is not limited to, the carbon-containing groups, residues or radicals defined above. Organic waste may contain several heteroatoms, or be linked to another molecule through a heteroatom, including oxygen, nitrogen, sulfur, phosphorus or the like. Examples of organic residues include, but are not limited to, substituted alkyl or alkyls, alkoxy or substituted alkoxy, mono or disubstituted amino groups, amide, and the like. Organic waste may preferably comprise 1 to 18 carbon atoms, 1 to 15 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms or 1 to 4 carbon atoms. In another aspect, an organic residue may comprise from 2 to 18 carbon atoms, from 2 to 15 carbon atoms, from 2 to 12 carbon atoms, from 2 to 8 carbon atoms, from 2 to 4 carbon atoms, or 2 at 4 carbon atoms.
A closely related synonym for the term "residue" is the term "radical" which, as used in the specification and the final claims, refers to a fragment, group or substructure of a molecule described herein, regardless of how The molecule is prepared. For example, a 2,4-thiazolidinedione radical in a given compound has the structure: regardless of whether thiazolidinedione is used to prepare the compound. In some embodiments, the radical (e.g. an alkyl) may be further modified (i.e. substituted alkyl) by having attached one or more "substituent radicals" to it. The number of atoms in a given radical is not critical to the present invention unless otherwise indicated elsewhere herein.
"Organic radicals," as the term is defined and used herein, contains one or more carbon atoms. An organic radical may have, for example, 1 to 26 carbon atoms, 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. . In another aspect, an organic radical may have 2 to 26 carbon atoms, 2 to 18 carbon atoms, 2 to 12 carbon atoms, 2 to 8 carbon atoms, 2 to 6 carbon atoms or 2 to 4 carbon atoms . Organic radicals usually have hydrogen bonded to at least some of the carbon atoms of the organic radical. An example of an organic radical that does not comprise inorganic atoms is a 5,6,7,8-tetrahydro-2-naphthyl radical. In some embodiments, an organic radical may contain from 1 to 10 inorganic heteroatoms attached to or within itself, including halogens, oxygen, sulfur, nitrogen, phosphorus and the like. Examples of organic radicals include, but are not limited to, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, monosubstituted amino, disubstituted amino, acyloxy, cyano, carboxy, carboalkoxy, alkylcarboxamide, substituted alkylcarboxamide, dialkylcarboxamide, alkylalkylsulfinyl, thioalkyl, thiohaloalkyl, alkoxy, substituted alkoxy, haloalkyl, haloalkoxy, aryl, substituted aryl, heteroaryl, heterocyclic or substituted heterocyclic radicals, wherein the terms are as defined elsewhere herein. Some non-limiting examples of organic radicals that include heteroatoms include alkoxy radicals, trifluoromethoxy radicals, acetoxy radicals, dimethylamino radicals and the like.
"Inorganic radicals", as the term is defined and used herein, do not contain carbon atoms and therefore comprise only different carbon atoms. Inorganic radicals comprise bonded combinations of selected atoms of hydrogen, nitrogen, oxygen, silicon, phosphorus, sulfur, selenium and halogens such as fluorine, chlorine, bromine and iodine, which may be present individually or in their chemically stable combinations. Inorganic radicals have 10 or less, or preferably one to six or one to four joined inorganic atoms, as indicated above. Examples of inorganic radicals include, but are not limited to, amino, hydroxyl, halogens, nitro, thiol, sulfate, phosphate and similar commonly known inorganic radicals. Inorganic radicals have not attached to them the metallic elements of the periodic table (such as alkali metals, alkaline earth metals, transition metals, lanthanide metals or actinide metals), although such metal ions can sometimes serve as a pharmaceutically acceptable cation. anionic inorganic radicals, such as a sulfate, phosphate, or similar anionic inorganic radicals. Inorganic radicals do not include metalloid elements such as boron, aluminum, gallium, germanium, arsenic, tin, lead or tellurium, or noble gas elements, unless specifically indicated elsewhere herein.
The compounds described herein may contain one or more double bonds and thus potentially yield cis / trans (E / Z) isomers as well as other conformational isomers. Unless otherwise defined, the invention includes all such possible isomers as well as mixtures of such isomers.
Unless otherwise defined, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, for example, each enantiomer and diastereoisomer, and a mixture of isomers as a racemic or scalemic mixture. The compounds described herein may contain one or more asymmetric centers and thus potentially give rise to diastereoisomers and optical isomers. Unless otherwise defined, the present invention includes all such possible diastereoisomers as well as racemic mixtures thereof, substantially pure separate enantiomers, all possible geometric isomers and pharmaceutically acceptable salts thereof. Mixtures of stereoisomers as well as specific isolated stereoisomers are also included. In the course of the synthetic procedures used to prepare such compounds, or using the racemization or epimerization procedures known to those skilled in the art, the products of such procedures may be a mixture of stereoisomers.
Many organic compounds exist in optically active forms, having the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule relative to its chiral / chiral center (s). The prefixes d e 1 or (+) and (-) are used to denote the rotation signal of the flat-polarized light of the compound, with (-) or 1 meaning that the compound is levogiro. A compound with prefix (+) or d is dextrogiro. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are non-overlapping specular images of each other. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often referred to as an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture.
Many of the compounds described herein may have one or more chiral centers and therefore may exist in different enantiomeric forms. If desired, a chiral carbon may be designated with an asterisk (*). When chiral carbon bonds are represented as straight lines in the disclosed formulas, it is understood that both (R) and (S) configurations of chiral carbon, and thus both enantiomers and mixtures thereof, are encompassed by the formula. As used in the art, when it is desired to specify the absolute configuration with respect to a chiral carbon, one of the bonds to the chiral carbon may be represented as a wedge (bonds to above plane atoms) and the other may be represented as a series. or wedge of small parallel lines (bonds to atoms below the plane). The Cahn-Inglod-Prelog system can be used to assign the (R) or (S) configuration of a chiral carbon.
[0100] The compounds described herein comprise atoms in their natural isotopic abundance and in unnatural abundance. The disclosed compounds may be isotopically labeled or isotopically substituted compounds which are identical to those described except that one or more atoms are replaced by an atom having atomic mass or mass number other than atomic mass or mass number generally. found in nature. Examples of isotopes which may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine such as 2H, 3H, 13C, 14C, 15N, 180, 170, 35S, 18F and 36 Cl. respectively. The compounds further comprising their prodrugs and pharmaceutically acceptable salts of said compounds or said prodrugs containing the above isotopes and / or other isotopes of the other atoms are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, for example, those in which radioactive isotopes, such as 3 H and 14 C, are incorporated, and useful in drug and / or substrate tissue distribution assays. Tritium (i.e. 3H) and carbon 14 (i.e. 14C) isotopes are particularly preferred for their ease of preparation and detection. In addition, substitution with heavier isotopes, such as deuterium, ie 2H, may provide certain therapeutic advantages resulting from increased metabolic stability, for example longer in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances. The isotopically labeled compounds of the present invention and prodrugs thereof may generally be prepared by performing the procedures below by replacing a readily available isotopically labeled reagent with an isotopically unlabeled reagent.
The compounds described in the invention may be present as a solvate. In some cases, the solvent used to prepare the solvate is an aqueous solution, and the solvent is then often referred to as a hydrate.
The compounds may be present as a hydrate, which may be obtained, for example, by crystallization from a solvent or aqueous solution. In this context, one, two, three or any arbitrary number of solvent or water molecules may combine with the compounds according to the invention to form solvates and hydrates. Unless otherwise defined, the invention includes all such possible solvates.
The term "cocrystal" means a physical association of two or more molecules whose stability occurs through non-covalent interaction. One or more components of this molecular complex provide a stable structure in the crystalline lattice. In certain cases, guest molecules are incorporated into the crystalline lattice as anhydrates or solvates, see, for example, "Crystal Engineering of the Composition of Pharmaceutical Phases. Do Pharmaceutical Co-crystals Represent a New Path to Improved Medicines " Almarasson, 0., et. al., The Royal Society of Chemistry, pages 1889 to 1896, 2004. Examples of cocrystals include p-toluenesulfonic acid and benzenesulfonic acid.
It is also estimated that certain compounds described herein may be present as a balance of tautomers. For example, ketones with an α-hydrogen may exist in a balance of keto form and enol form.
Similarly, amides with an N-hydrogen may exist in a balance of the amide form and the imidic acid form. Unless otherwise defined, the invention includes such possible tautomers.
Chemicals are known to form solids that are present in different states of order which are termed polymorphic forms or modifications. The different modifications of a polymorphic substance may differ greatly in their physical properties. The compounds according to the invention may be present in different polymorphic forms, and certain modifications may be metastable. Unless otherwise defined, the invention includes all such possible polymorphic forms.
In some aspects, a structure of a compound may be represented by a formula: which is understood to be equivalent to a formula: where n is normally an integer. That is, Rn is understood to represent five independent substituents, Rn (a), Rn (b), Rn (c), Rn (d) and Rn (e). By "independent substituents" is meant that each substituent R may be independently defined. For example, if in one example Rn <a> is halogen, then Rn <b> is not necessarily halogen in that example.
Certain materials, compounds, compositions and components disclosed herein may be obtained commercially or readily synthesized using techniques generally known to those skilled in the art. For example, starting materials and reagents used in the preparation of the disclosed compounds and compositions are available from commercial suppliers such as Sigma-Aldrich Chemical Co. (Milwaukee, WI), Acros Organics (Morris Plains, NJ), Fisher Scientific (Pittsburgh, PA.) Or Sigma (St. Louis, MO.), Or are prepared by methods known to those skilled in the art according to the procedures defined in the references, such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1. 17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1 to 5 and Supplements (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1 to 40 (John Wiley and Sons, 1991); March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition); and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989).
Unless expressly indicated, it is by no means intended that any method defined in this document be construed as requiring that its steps be performed in a specific order. In that sense, where a method claim does not effectively cite an order to be followed by its steps or is not otherwise specifically defined in the claims or descriptions that the steps must be limited to a specific order, it is not intended to be no, that an order be inferred in any respect. This goes for any possible unspoken basis for interpretation, including questions of logic regarding the arrangement of steps or operational flow, the simple meaning derived from grammatical organization or punctuation; and the number or type of modalities described in the descriptive report.
The components to be used to prepare the compositions of the invention are disclosed, as well as the compositions themselves to be used according to the methods disclosed herein. These and other materials are disclosed in this document, and it is understood that when combinations subsets, interactions, groups, etc. Such materials are disclosed that while specific reference to each of the various individual and collective combinations and permutation of such compounds cannot 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 may be made to various molecules, including the compounds, are discussed, each and every combination and permutation of the compound and the modifications that are possible are specifically contemplated. unless specifically stated otherwise. 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, AD is disclosed, then even if each is not individually cited, each is individually and collectively contemplated, meaning that the AE, AF, BD, BE, BF, CD, CE and CF combinations are considered disclosed. Similarly, any subset or combination thereof is also disclosed. Thus, for example, the subgroup with A-E, B-F and C-E would be considered disclosed. This concept applies to all aspects of the present application including, but not limited to, the steps in the methods of making and using the compositions of the 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 invention.
[0110] It is understood that the compositions disclosed in this document have certain functions. Certain structural requirements for performing the disclosed functions are disclosed, and it is understood that there are a variety of structures that can perform the same function as they are related to the disclosed structures, and that these structures will usually achieve the same result.
B. COMPOUNDS
In one aspect, the invention relates to compounds useful as histone demethylase inhibitors. In another aspect, the compounds are useful as specific lysine histone demethylase ("LSD") inhibitors. Furthermore, in one aspect, the compounds of the invention are useful in the treatment of uncontrolled cell proliferation disorders. In another aspect, the disorder of uncontrolled cell proliferation is a cancer or a tumor. In yet a further aspect, the uncontrolled cell proliferation disorder is associated with an LSD dysfunction, as further described herein.
It is envisaged that each disclosed derivative may be optionally additionally substituted. It is also contemplated that any one or more derivative (s) may optionally be omitted from the invention. It is understood that a disclosed compound can be obtained by the disclosed methods. It is also understood that the disclosed compounds may be used in the disclosed methods of use.
1. STRUCTURE
In one aspect, the invention relates to a compound having a structure represented by formula (I) or formula (II): wherein m is 0 or 1; n is an integer from 0 to 3; X is selected from the group consisting of Z is selected from the group consisting of N and CH;
R1 is selected from the group consisting of halogen, C1-C3 haloalkyl and C1-C3 polyhaloalkyl; R 2, R 3 and R 4 are each independently selected from the group consisting of hydrogen, halogen, hydroxyl, cyano, amino, C 2 -C 6 alkoxy, C 1 -C 6 alkoxy, C 1 -C 6 alkyl, C 1 -C 6 polyhaloalkyl and C 1 haloalkyl. Cê;
Rs is selected from the group consisting of NR6R7, C1-C6 alkyl, C3-C6 cycloalkyl, and Cy, and is substituted with 0 to 3 groups independently selected from halogen, hydroxyl, amino, C2-C6 alkoxy, C1-C6 alkyl alcohol, alkoxy C1-C6, C1-C6 alkyl, C1-C6 polyhaloalkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl and Cy;
Cy is a heterocycloalkyl selected from the group consisting of aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, oxazolidinyl, imidazolidinyl, pyrazolidinyl, piperazinyl, oxazinanyl, morpholinyl, hexahydropyrimidinyl and hexahydropyridine; and each of R 8 and R 7 is independently selected from the group consisting of hydrogen, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl and C 3 -C 6 heterocycloalkyl; or a pharmaceutically acceptable salt thereof.
In some embodiments, the invention provides a compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof.
In still other embodiments, the invention provides a compound having a structure represented by a formula: or a pharmaceutically acceptable salt thereof.
In still other embodiments, the invention provides a compound having a structure represented by a formula: or a pharmaceutically acceptable salt thereof.
The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of any compound of the invention and a pharmaceutically acceptable carrier.
The invention also provides a method for treating an uncontrolled cell proliferation disorder 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 invention also provides a method for decreasing histone demethylase activity in a mammal, the method comprising the step of administering to the mammal an effective amount of any of the compounds of the invention.
2. INHIBITION OF HISTON DEMETILASE ACTIVITY
In one aspect, the disclosed compounds exhibit inhibition of LSD protein activity. In a still further aspect, the disclosed compounds exhibit selective inhibition of LSD1 protein activity. In yet another additional aspect, the disclosed compounds exhibit selective inhibition of LSD2 protein activity. In yet another aspect, the disclosed compounds inhibit LSD demethylase activity. In yet another aspect, the disclosed compounds exhibit binding to the LSD FAD domain. In yet another additional aspect, the disclosed compounds exhibit inhibition of histone 3 (H3) LSD-mediated demethylation at the Lys4 position. In yet another aspect, the disclosed compounds exhibit inhibition of H3K3ml and H3K4me2 LSD-mediated demethylation. In yet another aspect, the disclosed compounds exhibit inhibition of H3K9me2 and H3K9mel LSD-mediated demethylation.
In yet another aspect, the disclosed compounds inhibit LSD1 demethylase activity. In yet another aspect, the disclosed compounds exhibit binding to the LSD1 FAD domain. In yet another additional aspect, the disclosed compounds exhibit inhibition of histone 3 (H3) LSD1 mediated demethylation at the Lys4 position. In yet another aspect, the disclosed compounds exhibit inhibition of H3K3ml and H3K4me2 LSD1-mediated demethylation. In yet another aspect, the disclosed compounds exhibit inhibition of H3K9me2 and H3K9mel LSD1 mediated demethylation.
In yet another aspect, the disclosed compounds inhibit LSD2 demethylase activity. In yet another aspect, the disclosed compounds exhibit binding to the LSD2 FAD domain. In yet another additional aspect, the disclosed compounds exhibit inhibition of histone 3 (H3) LSD2 mediated demethylation at the Lys4 position. In yet another aspect, the disclosed compounds exhibit inhibition of H3K3ml and H3K4me2 LSD2 mediated demethylation.
In yet another aspect, the disclosed compounds exhibit disruptions of LSD interaction with a complex comprising one or more of the HDAC1 / 2, CoREST, CtBP1, BRAF35 and BHC80 proteins. In yet another aspect, the disclosed compounds disrupt LSD1 binding to one or more proteins selected from the HDAC1 / 2, CoREST, CtBP1, BRAF35 and BHC80 proteins. In yet another additional aspect, the disclosed compounds disrupt LSD2 binding to one or more proteins selected from G9a, NSD3, HDAC1 / 2, CoREST, CtBP1, BRAF35 and BHC80 proteins.
Inhibition of LSD activity can be determined by a variety of in vitro and in vivo methods known to one of ordinary skill in the art. For example, enzymatic activity may be determined in in vitro enzyme assay systems. In many respects, the enzymatic activity of LSD1 or LSD2 can be determined in a spectrophotometric assay. In summary, the assay is based on the multi-step enzymatic reaction in which LSD1 or LSD2 primarily produces H2O2 during demethylation of lysine 4 into a peptide corresponding to the first 21 amino acids of the histone H3 N-terminal tail. In the presence of horseradish peroxidase, the produced H2O2 reacts with ADHP to produce the highly fluorescent resorufin compound which can be analyzed with an excitation wavelength of 530 to 540 nm and an emission wavelength of 585 to 595 nm. The assay requires an LSD1 or LSD2 enzyme source, purified from natural sources (e.g., from cultured tissue or cells), isolated as a recombinantly expressed protein or as an impure protein in whole cell extracts. In one aspect, the disclosed compounds exhibit inhibition of LSD protein activity with an IC50 in an EMSA assay less than about 300 μΜ, less than about 100 μΜ, less than about 50μΜ, less than about 10 μΜ, less than about 1 μΜ, less than about 500 nM or less than about 100 nM. In another aspect, the disclosed compounds exhibit inhibition of LSD1 protein activity with an IC50 in an EMSA assay less than about 300 μΜ, less than about 100 μΜ, less than about 50 M, less than about 10 μΜ, less than about 1 μΜ, less than about 500 nM or less than about 100 nM. In yet another aspect, the disclosed compounds exhibit inhibition of LSD2 protein activity with an IC50 in an EMSA assay less than about 300 μΜ, less than about 100 μΜ, less than about 50 μΜ, less than about 10 μΜ , less than about 1 μΜ, less than about 500 nM or less than about 100 nM.
In one aspect, the disclosed compounds are selective for LSD. In another aspect, selective inhibition of LSD activity is determined using an enzyme assay. In several other respects, the compound inhibits LSD activity in an enzyme assay with an IC50 lower than IC50 for MAO A and / or MAO B. That is, a disclosed compound may have selectivity for LSD protein vis-à-vis vis MAO A and / or MAO B. For example, in one aspect, a disclosed compound may inhibit LSD with an IC50 about 5 times lower than that of MAO A, about 10 times lower than that of MAO A, about 20 times smaller than that of MAO A, about 30 times smaller than that of MAO A, about 50 times smaller than that of MAO A, about 100 times smaller than that of MAO A, about 250 times smaller than MAO A, about 500 times smaller than MAO A, about 1000 times smaller than MAO A, and more than about 1000 times smaller than MAO A In another aspect, a disclosed compound may inhibit LSD with an IC 50 about 5 times lower than that of MAO B, about 10 times lower than that of MAO B, about 20 times lower than that of MAO B about 30 times m less than that of MAO B, about 50 times smaller than that of MAO B, about 100 times smaller than that of MAO B, about 250 times smaller than that of MAO B, about 500 times smaller than than that of MAO B, about 1000 times smaller than that of MAO B and more than about 1000 times smaller than that of MAO B.
In one aspect, the disclosed compounds are selective for LSD1. In another aspect, selective inhibition of LSD1 activity is determined using an enzyme assay.
In several additional aspects, the compound inhibits LSD1 activity in an enzyme assay with an IC50 lower than IC50 for one or more of the LSD2, MAO A, and MAO B enzymes. That is, a disclosed compound may have selectivity for the LSD1 protein vis-à-vis One or more of LSD2, MAO A and MAO B. For example, in one aspect, a disclosed compound may inhibit LSD1 with an IC50 of about 5 times lower than LSD2, about 10 times lower. than LSD2, about 20 times smaller than LSD2, about 30 times smaller than LSD2, or about 50 times smaller than LSD2. In another aspect, a disclosed compound may inhibit LSD1 with an IC 50 about 5 times lower than that of MAO A, about 10 times lower than that of MAO A, about 20 times lower than that of MAO A about 30 times smaller than that of MAO A, about 50 times smaller than that of MAO A, about 100 times smaller than that of MAO A, about 250 times smaller than that of MAO A, about 500 times smaller than that of MAO A, about 1000 times smaller than that of MAO A, and more than about 1000 times smaller than that of MAO A. In another aspect, a disclosed compound may inhibit LSD1 with an IC50 about 5 times lower than that of MAO B, about 10 times lower than that of MAO B, about 20 times lower than that of MAO B, about 30 times lower than that of MAO B, about 50 times smaller than that of MAO B, about 100 times smaller than that of MAO B, about 250 times smaller than that of MAO B, about 500 times smaller than that of MAO B, about 1000 times smaller than da B and more than q which is about 1000 times smaller than that of B.
In one aspect, the disclosed compounds are selective for LSD2. In another aspect, selective inhibition of LSD2 activity is determined using an enzyme assay.
In several additional aspects, the compound inhibits LSD2 activity in an enzyme assay with an IC50 lower than IC50 for one or more of the LSD1 enzymes, A and B. That is, a disclosed compound may have selectivity for the LSD2 protein vis-à-vis One or more of LSD1, A and B. For example, in one aspect, a disclosed compound may inhibit LSD2 with an IC50 of about 5 times lower than that of LSD1, about 10 times lower. than LSD1, about 20 times smaller than LSD1, about 30 times smaller than LSD1, or about 50 times smaller than LSD1. In another aspect, a disclosed compound may inhibit LSD2 with an IC50 about 5 times lower than that of A, about 10 times lower than that of A, about 20 times lower than that of A , about 30 times smaller than that of A, about 50 times smaller than that of A, about 100 times smaller than that of A, about 250 times smaller than that of A, about 500 times smaller than that of A, about 1000 times smaller than that of A, and more than about 1000 times smaller than that of A. In another aspect, a disclosed compound can inhibit LSD2 with an IC50 about 5 times lower than that of MAO B, about 10 times lower than that of MAO B, about 20 times lower than that of MAO B, about 30 times lower than that of MAO B, about 50 times smaller than that of MAO B, about 100 times smaller than that of MAO B, about 250 times smaller than that of MAO B, about 500 times smaller than that of MAO B, about 1000 times smaller than that of MAO B is more than about 1000 times smaller than that of MAO B.
In various aspects, the disclosed compounds exhibit binding to an LSD protein. In another aspect, the disclosed compounds exhibit binding to the FAD domain of an LSD protein. In yet another aspect, the disclosed compounds exhibit LSD1 protein binding. In yet another additional aspect, the disclosed compounds exhibit LSD2 protein binding. The binding affinity of a disclosed compound for an LSD protein, for example LSD1 protein, can be determined by various methods known to one of ordinary skill in the art. In one aspect, the disclosed compounds exhibit LSD protein binding with a Kd of less than about 50 μΜ, less than about 10 μΜ, less than about 1 μΜ, less than about 500 nM or less than about 100 nM. In another aspect, Kd is determined using an SPR method. In yet another aspect, binding is determined using the LSD1 protein. In yet a further aspect, binding is determined using the LSD2 protein.
[0129] In many additional respects, LSD binding is selective. In a further aspect, the disclosed compounds exhibit a lower LSD binding Kd than the MAO A and / or MAO B KD. That is, a disclosed compound may have selectivity for the LSD protein vis-à-vis MAO A and / or MAO B proteins. For example, in one aspect, a disclosed compound may bind LSD with a Kd about 5 times lower than that of MAO A, about 10 times lower than that of MAO A, about 20 times smaller than that of MAO A, about 30 times smaller than that of MAO A, about 50 times smaller than that of MAO A, about 100 times smaller than that of MAO A, about 250 times smaller than MAO A, about 500 times smaller than MAO A, about 1000 times smaller than MAO A, and more than about 1000 times smaller than MAO A In another aspect, a disclosed compound may bind LSD with a Kd about 5 times lower than that of MAO B, about 10 times lower than that of MAO B, about 20 times lower than that of MAO. B, about 30 times m less than that of MAO B, about 50 times smaller than that of MAO B, about 100 times smaller than that of MAO B, about 250 times smaller than that of MAO B, about 500 times smaller than than that of MAO B, about 1000 times smaller than that of MAO B and more than about 1000 times smaller than that of MAO B.
[0130] In many additional respects, the binding of LSD1 is selective. In another aspect, the disclosed compounds exhibit a lower Kd for LSD1 binding than one Kd for one or more of the LSD2, MAO A and MAO B. That is, a disclosed compound may have selectivity for the LSD1 protein vis-à-vis vis one or more of the LSD2, MAO A and MAO B proteins. For example, in one aspect, a disclosed compound may bind LSD1 with a Kd of about 5 times lower than that of LSD2, about 10 times lower than than LSD2, about 20 times smaller than LSD2, about 30 times smaller than LSD2, or about 50 times smaller than LSD2. In another aspect, a disclosed compound may bind LSD1 with a Kd about 5 times lower than that of MAO A, about 10 times lower than that of MAO A, about 20 times lower than that of MAO A, about 30 times smaller than that of MAO A, about 50 times smaller than that of MAO A, about 100 times smaller than that of MAO A, about 250 times smaller than that of MAO A, about 500 times smaller than that of MAO A, about 1000 times smaller than that of MAO A, and more than about 1000 times smaller than that of MAO A. In another aspect, a disclosed compound may bind to LSD1 with a Kd about 5 times lower than that of MAO B, about 10 times lower than that of MAO B, about 20 times lower than that of ΜΑΟ B, about 30 times lower than that of ΜΑΟ B, about 50 times smaller than that of B, about 100 times smaller than that of B, about 250 times smaller than that of B, about 500 times smaller than that of B, about 1000 times smaller than that of ΜΑΟ B and more than about 1000 times smaller than that of ΜΑΟ B.
[0131] In many additional respects, LSD2 binding is selective. In another aspect, the disclosed compounds exhibit a lower Kd for LSD2 binding than one Kd for one or more of the LSD1, ΔA and ΔB. That is, a disclosed compound may have selectivity for the LSD2 protein vis-à-vis vis one or more of the LSD1 proteins, A and B. For example, in one aspect, a disclosed compound can bind LSD2 with a Kd of about 5 times lower than that of LSD1, about 10 times lower than than LSD1, about 20 times smaller than LSD1, about 30 times smaller than LSD1, or about 50 times smaller than LSD1. In another aspect, a disclosed compound may bind LSD2 with a Kd about 5 times lower than that of ΜΑΟ A, about 10 times lower than that of ΜΑΟ A, about 20 times less than that of ΜΑΟ A, about 30 times smaller than that of MAO A, about 50 times smaller than that of MAO A, about 100 times smaller than that of MAO A, about 250 times smaller than that of MAO A, about 500 times smaller than that of MAO A, about 1000 times smaller than that of MAO A, and more than about 1000 times smaller than that of MAO A. In another aspect, a disclosed compound may bind to LSD2 with a Kd about 5 times lower than that of MAO B, about 10 times lower than that of MAO B, about 20 times lower than that of MAO B, about 30 times lower than that of MAO B, about 50 times smaller than that of MAO B, about 100 times smaller than that of MAO B, about 250 times smaller than that of MAO B, about 500 times smaller than that of MAO B, about 1000 times smaller than that of MAO B in more than about 1000 times smaller than that of MAO B.
Alternatively, inhibition of STAT protein activity may be determined in a cell-based assay. There are a variety of cell-based assays that are suitable for determining inhibition of LSD protein activity known to one of ordinary skill in the art. For example, inhibition of cell growth or cell disruption can be determined using a cell, either a permanent cell line or a primary cell culture that has a dysfunctioning LSD protein. In another aspect, the LSD protein is LSD1. In yet another aspect, the LSD protein is LSD2. In an additional aspect, LSD protein dysfunction is one in which the LSD protein has acquired a mutation gain in function. Alternatively, LSD protein dysfunction has a persistent or constitutive activity phenotype. For example, the LSD protein may have persistent or constitutive activity due to a dysfunction in an upstream regulatory protein. In another aspect, the LSD protein is overexpressed due to a dysfunction in LSD gene transcription and / or translation regulation. In another aspect, the cell houses an active oncogene that is associated with LSD dysfunction.
In one aspect, the disclosed compounds and the products of the disclosed methods of inhibiting cell growth. In yet another aspect, the disclosed compounds and the products of the disclosed methods inhibit cell growth in an in vitro assay system. In yet another aspect, the in vitro assay system makes use of a cell line derived from a cancer or tumor selected from breast cancer, ovarian cancer, testicular cancer, lung cancer, liver cancer, prostate cancer, breast cancer. pancreas and a sarcoma. In yet a further aspect, the cell line is derived from a human source. In a still further aspect, the disclosed compounds inhibit cell growth in a cell with a persistently active LSD protein. In yet another aspect, the cell line has an activated LSD protein. In a still further aspect, the cell line is selected from 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. In one aspect, the disclosed compounds exhibit inhibition of cell growth activity in an in vitro cell-based assay with an IC50 of less than about 500 μΜ, less than about 250 μΜ, less than about 100 μΜ, less than about 50 μΜ, less than about 10 μΜ, less than about 1 μΜ, less than about 500 nM, less than about 100 nM, less than about 10 nM, and less than about 1 nM.
In one aspect, the disclosed compounds and the products of the disclosed methods of inhibiting cell migration. In yet another aspect, the disclosed compounds and the products of the disclosed methods inhibit cell migration in an in vitro assay system. In yet another aspect, the in vitro assay system makes use of a cell line derived from a cancer or tumor selected from breast cancer, ovarian cancer, testicular cancer, lung cancer, liver cancer, prostate cancer, breast cancer. pancreas and a sarcoma. In yet a further aspect, the cell line is derived from a human source. In a still further aspect, the disclosed compounds inhibit cell growth in a cell with a persistently active LSD protein. In yet another aspect, the cell line has an activated LSD protein. In a still further aspect, the cell line is selected from 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. In one aspect, the disclosed compounds exhibit inhibition of cell migration in an in vitro cell-based assay with an IC50 of less than about 300 μΜ, less than about 100 μΜ, less than about 50μΜ, less than about 10 μΜ , less than about 1 μΜ, less than about 500 nM or less than about 100 nM.
C. METHODS FOR MAKING COMPOUNDS
In one aspect, the invention relates to methods for making compounds useful as LSD inhibitors. In another aspect, the products of the disclosed manufacturing methods are modulators of LSD's activity. In yet another aspect, the products of the disclosed methods of binding to a STAT protein and negatively modulating LSD activity. The compounds may, in one aspect, exhibit subtype selectivity. In yet another aspect, the products of the disclosed methods present selectivity for the LSD1 member of the LSD protein family. In yet another aspect, the products of the disclosed methods of presenting selectivity for the LSD2 member of the LSD protein family.
In one aspect, the invention relates to methods for making compounds useful as histone demethylase inhibitors, which may be useful in the treatment of uncontrolled cell proliferation disorders. In another aspect, the histone demethylase is LSD1. In a still further aspect, the histone demethylase is LSD2.
[0137] The compounds of this invention may be prepared using reactions as shown in the following reaction schemes, in addition to other standard manipulations which are known in the literature or exemplified in the experimental or clear sections for one skilled in the art. For the sake of clarity, single substituent examples are shown where multiple substituents are permitted under the definitions disclosed herein.
[0138] The reactions used to generate the compounds of this invention are prepared using reactions as shown in the following reaction schemes, in addition to other standard manipulations that are known in the literature or to one skilled in the art. The following examples are provided so that the invention may be more fully understood, are illustrative only and should not be construed as limiting.
In one aspect, the disclosed compounds comprise the products of the synthetic methods disclosed therein. In another aspect, the disclosed compounds comprise a compound produced by a synthetic method described herein. In a still further aspect, the invention comprises a pharmaceutical composition comprising a therapeutically effective amount of the product of the disclosed methods and a pharmaceutically acceptable carrier. In yet another aspect, the invention comprises a method of manufacturing a medicament comprising combining at least one compound of any of the disclosed compounds or at least one product of the disclosed methods with a pharmaceutically acceptable carrier or diluent.
1. ROUTE I
In one aspect, (E) -N '- (1-phenylethylidene) benzohydrazide analogs of the present invention may be prepared generically by the synthetic scheme as shown below.
The compounds are represented generically with substituents as noted in the compound descriptions elsewhere herein. A more specific example is defined below.
In one aspect, Route I begins with a suitable substituted acid derivative (1.1). Suitable substituted acid derivatives (1.1) are commercially available or may be readily prepared by one of ordinary skill in the art. In a typical reaction, the type 1.1 compound is added to the type 1.2 amine derivative in the presence of a suitable base, for example potassium carbonate, in a suitable solvent such as THF. The reaction is stirred at room temperature (about 15 to 30 ° C) for a period of time sufficient to complete the reaction, for example about twelve hours. After completion of the reaction, the solvent is removed under vacuum, and the type 1.3 compound is isolated and purified by chromatography.
[0143] In one aspect, type 1.4 compounds may be prepared by reacting type 1.3 compounds with an alcohol by an esterification reaction. In a typical reaction, a type 1.3 compound is heated to a suitable temperature (for example under reflux at about 65 ° C) in a suitable alcoholic solvent, for example methanol in the presence of an acid catalyst such as concentrated sulfuric acid. for a sufficient time to complete the reaction, for example overnight (about 8 to 18 h). Upon completion of the reaction, the solvent is removed under vacuum, and the type 1.4 compound is isolated and purified by chromatography.
In one aspect, type 1.4 compounds may provide type 1.5 compounds by reaction with an appropriate hydrazine derivative (NH2NHR4). In a typical reaction, a type 1.4 compound is added to a suitable hydrazine derivative (NH2NHR4) and heated to a suitable temperature (e.g. under reflux at about 65 ° C) in a suitable solvent, eg methanol, for a period of time sufficient to complete the reaction (eg about 12 h). Upon completion of the reaction, the solvent is removed under vacuum, and the type 1.5 compound is isolated and purified by chromatography.
[0145] In one aspect, type 1.5 compounds may provide type 1.7 compounds by reaction with an appropriate carbonyl containing compound (1.6). In a typical reaction, a type 1.6 compound and an appropriate hydrazine derivative (1.5) are dissolved in a suitable solvent, for example methanol, in the presence of a suitable acid catalyst (eg acetic acid), and the mixture is heated. using a microwave reactor at a suitable temperature, for example about 120 ° C, for a period of time sufficient to complete the reaction (eg about 30 min). After completion of the reaction and cooling, the solvent is removed under vacuum and type 1.7 compounds are isolated and purified by chromatography.
2. ROUTE II
In one aspect, (E) -N '- (1-phenylethylidene) benzohydrazide analogs of the present invention may be prepared generically by the synthetic scheme as shown below.
The compounds are represented generically, with substituents, as noted in the compound descriptions elsewhere herein. A more specific example is defined below.
In one aspect, Route II begins with a suitable substituted acid derivative (2.1). Suitable substituted acid derivatives (2.1) are commercially available or can be readily prepared by one skilled in the art. In one aspect, type 2.2 compounds may be prepared by reacting type 2.1 compounds with an alcohol by an esterification reaction. In a typical reaction, a type 2.1 compound is heated to a suitable temperature (for example under reflux at about 70 ° C) in a suitable alcoholic solvent, for example methanol in the presence of an acid catalyst such as concentrated sulfuric acid. for a period of time sufficient to complete the reaction, for example overnight (about 8 to 18 h). Upon completion of the reaction, the solvent is removed under vacuum, and the type 2.2 compound is isolated and purified by chromatography.
In one aspect, type 2.2 compounds may provide type 2.3 compounds by reaction with an appropriate hydrazine derivative (NH2NHR4). In a typical reaction, a type 2.2 compound is added to a suitable hydrazine derivative (NH2NHR4) and heated to a suitable temperature (e.g. under reflux at about 70 ° C) in a suitable solvent, eg methanol, long enough to complete the reaction, such as overnight (for example, 8 to 18 hours). Upon completion of the reaction, the solvent is removed under vacuum, and the type 2.3 compound is isolated and purified by chromatography.
[0150] In one aspect, type 2.3 compounds may be used to produce type 2.5 compounds by reaction with an appropriate carbonyl containing compound (2.4). In a typical reaction, a 2.4 compound and an appropriate hydrazine derivative (2.3) are dissolved in a suitable solvent, for example methanol, in the presence of a suitable acid catalyst (eg acetic acid), and the mixture is heated using a microwave reactor at a suitable temperature, for example about 120 ° C, for a period of time sufficient to complete the reaction (eg about 30 min). After completion of the reaction and cooling, the solvent is removed under vacuum and type 2.5 compounds are isolated and purified by chromatography.
In a further aspect, the compound produced exhibits inhibition of a histone demethylase. In another additional aspect, histone demethylase is a member of the specific lysine family ("LSD") of histone demethylases. In a still further aspect, the histone demethylase is LSD1. In a still further aspect, the histone demethylase is LSD2. In a still further aspect, the compound produced exhibits inhibition of cell viability.
In a further aspect, the produced compound exhibits inhibition with an IC50 of less than about 1.0 ΧΙΟ-4M. In yet another aspect, the produced compound exhibits inhibition with an IC50 of less than about 1.0 ΧΙΟ. -5M. In yet a further aspect, the produced compound exhibits inhibition with an IC50 of less than about 1.0 ΧΙΟ -6M. In yet another aspect, the produced compound exhibits inhibition with an IC50 of less than about 1, In an additional aspect, the produced compound exhibits inhibition with an IC 50 of less than about 1.0 ΧΙΟ -8 M. In yet another aspect, the produced compound exhibits inhibition with an IC 50 of less than about 1.0 ΧΙΟ-9 M.
It is envisaged that each of the disclosed methods may further comprise additional steps, manipulations, and / or components. It is also contemplated that any one or more steps, manipulations and / or components may be optionally omitted from the invention. It is understood that disclosed methods may be used to produce the disclosed compounds. It is also understood that the products of the disclosed methods may be used in the disclosed methods of use.
D. PHARMACEUTICAL COMPOSITIONS
In one aspect, the invention relates to pharmaceutical compositions comprising the disclosed compounds. That is, a pharmaceutical composition may be provided, comprising a therapeutically effective amount of at least one disclosed compound or at least one product of a disclosed method, and a pharmaceutically acceptable carrier.
In a further aspect, the invention relates to pharmaceutical compositions comprising a pharmaceutically acceptable carrier and an effective amount of the product of a disclosed synthetic method. In a further aspect, the effective amount is a therapeutically effective amount. In another 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 salt / salts thereof as an active ingredient, a pharmaceutically acceptable carrier and optionally other therapeutic ingredients or adjuvants). The present compositions 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 for which it is present. the active ingredient is being administered The pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical art.
As used herein, the term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When the compound of the present invention is acidic, its corresponding salt may conveniently be prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (cupric and cuprous), ferric, earthy, lithium, magnesium, manganese (manganic and manganous), potassium, sodium, zinc and similar salts. Particularly preferred salts are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable non-toxic bases include salts of primary, secondary and tertiary amines, as well as cyclic amines and substituted amines, such as synthesized naturally occurring and substituted amines. Other pharmaceutically acceptable organic non-toxic bases from which salts may be formed include ion exchange resins such as arginine, betaine, caffeine, choline, N, 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, trimethylamine, trimethylamine, trimethylamine .
As used herein, the term "pharmaceutically acceptable non-toxic acids" includes inorganic acids, organic acids and prepared salts thereof, for example acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic acids , glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic and the like. Preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric and tartaric acids.
In practice, the compounds of the invention, or pharmaceutically acceptable salts thereof thereof may be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical composition techniques. The carrier may take a variety of forms depending upon the desired preparation form for administration, for example 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, wafers or tablets, each containing a predetermined amount of the active ingredient. In addition, the compositions may be presented as a powder, as granules, as a solution 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 common dosage forms set forth above, the compounds of the invention and / or pharmaceutically acceptable salt / salts thereof may also be administered by controlled release means and / or delivery devices. The compositions may be prepared by any of the pharmaceutical methods. In general, such methods include a step of associating the active ingredient with the carrier which constitutes one or more required ingredients. In general, the compositions are prepared by uniformly and intimately mixing the active ingredient with liquid carriers, solid or finely divided carriers or both. Then the product can be conveniently molded to the desired presentation.
Thus, the pharmaceutical compositions of this invention may include a pharmaceutically acceptable carrier and a compound or a pharmaceutically acceptable salt of the compounds of the invention. The compounds of the 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 vehicles are sugar syrup, peanut oil, olive oil and water. Examples of gaseous vehicles include carbon dioxide and nitrogen.
In preparing the compositions for oral dosage formulation, any covalent pharmaceutical medium may be used. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like may be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like may be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease in administration, tablets and capsules are the preferred oral dosage units by which solid pharmaceutical carriers are used. Optionally, the tablets may be coated by standard aqueous or non-aqueous techniques.
A tablet containing the composition of this invention may be prepared by compression or molding, optionally with one or more adjuvant or adjuvant ingredients. Compressed tablets may be prepared by compression, in a suitable machine, the active ingredient in a free flowing form, such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surfactant or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
The pharmaceutical compositions of the present invention comprise a compound of the invention (or pharmaceutically acceptable salts thereof) as an active ingredient, a pharmaceutically acceptable carrier and optionally one or more additional therapeutic agents or adjuvants. The present compositions include compositions suitable for oral, rectal, topical and parenteral administration (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 it is present. The active ingredient is being administered. The pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical art.
Pharmaceutical compositions of the present invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant may be included, for example hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. In addition, a preservative may be included to prevent harmful growth of microorganisms.
Pharmaceutical compositions of the present 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 effectively fluid for easy syringability. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; therefore, they should preferably be protected against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils and suitable mixtures thereof.
[0167] The pharmaceutical compositions of the present invention may be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, talc, mouthwash, gargling and the like. In addition, the compositions may be in a form suitable for use in transdermal devices. Such formulations may be prepared using a compound of the invention, or pharmaceutically acceptable salts thereof, by conventional processing methods. For example, a cream or ointment is prepared by mixing hydrophilic material and water, together with about 5 wt.% To about 10 wt.% Of the compound, to produce a cream or ointment of desired consistency.
[0168] The pharmaceutical compositions of this invention may be in a form suitable for rectal administration, wherein the carrier is a solid. It is preferable for the mixture to form unit dosage suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories may conveniently be formed primarily by mixing the composition with the molten or softened carrier (s), followed by refrigeration and molding in the molds.
In addition to the above-mentioned carrier ingredients, the pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants) and the like. In addition, other adjuvants may be included to make the formulation isotonic with the intended recipient's blood. Compositions containing a compound of the invention, and / or pharmaceutically acceptable salts thereof, may also be prepared in concentrated powder or liquid form.
Under treatment conditions requiring inhibition or negative modulation of LSD protein activity, an appropriate dosage level will generally be about 0.01 to 500 mg per kg body weight of the patient per day, and may be administered. in single or multiple doses. Preferably, the dosage level will be from about 0.1 to about 250 mg / kg per day; more preferably from 0.5 to 100 mg / kg per day. A suitable dosage level may be about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range, the dosage may be 0.05 to 0.5, 0.5 to 5.0, or 5.0 to 50 mg / kg per day. For oral administration, the compositions are preferably provided as tablets containing 1.0 to 1000 milligrams of active ingredient, particularly 1, 0, 5, 0, 10, 15, 20, 25, 50, 75, 100, 150, 200 , 250, 300, 400, 500, 600, 750, 800, 900 and 1000 milligrams of the active ingredient for symptomatic dose adjustment of the patient to be treated. The compound may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosage regimen can be adjusted to provide the optimal therapeutic response.
It is understood, however, that the specific dose level for any particular patient will depend on a variety of factors. Such factors include the age, body weight, general health, gender and diet of the patient. Other factors include time and route of administration, excretion rate, drug combination, and the type and severity of specific disease on therapy.
[0172] The present invention is further directed to a method for the manufacture of a medicament for inhibiting or negatively modulating LSD protein activity (e.g., treating an uncontrolled cell proliferation disorder, or one or more neurodegenerative disorders associated with LSD dysfunction) in mammals (e.g., humans), comprising combining one or more disclosed compounds, products or compositions with a pharmaceutically acceptable carrier or diluent. Thus, in one aspect, the invention relates to a method of manufacturing 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 may be prepared from the disclosed compounds. It is also understood that the disclosed compositions may be used in the disclosed methods of use.
E. METHODS OF USE OF COMPOUNDS AND COMPOSITIONS
The disclosed compounds may be used as individual agents or in combination with one or more other drugs in the treatment, prevention, control, amelioration or reduction of the risk of the aforementioned diseases, disorders and conditions for which the compounds of formula I or other drugs have utility, where the combination of drugs together is safer or more effective than any drug alone. The other drug (s) may be administered by one route and in an amount commonly used for that purpose, simultaneously or sequentially with a disclosed compound. When a disclosed compound is used simultaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compound is preferred. However, combination therapy may also be administered in overlapping schedules. It is also anticipated that the combination of one or more active ingredients and a disclosed compound will be more effective than either as an individual agent.
The pharmaceutical compositions and methods of the present invention may further comprise other therapeutically active compounds, as observed in the present invention, which are generally applied in the treatment of the above-mentioned pathological conditions.
1. TREATMENT METHODS
The compounds disclosed in the present invention are useful for treating, preventing, ameliorating, controlling or reducing the risk of a variety of disorders, in which the patient or individual could benefit from the inhibition or negative modulation of a protein. LSD. In one aspect, a treatment may include selective inhibition of LSD to an effective extent to affect histone demethylation activity. Thus, a disorder may be associated with histone demethylation activity, for example, with dysfunctional epigenetic regulation of genes in a cancer cell. In one aspect, there is provided a method of treating or preventing a disorder in an individual comprising the step of administering to the individual at least one disclosed composition; and / or at least one disclosed pharmaceutical composition; and / or at least one product disclosed in a dosage and amount effective to treat the disorder in the subject.
Also provided is a method for treating one or more disorders, for which inhibition of LSD is expected to be beneficial in an individual, comprising the step of administering to the individual at least one disclosed compound; at least one disclosed pharmaceutical composition; and / or at least one product disclosed in a dosage and amount effective to treat the disorder in the subject.
In one aspect, there is provided a method for treating an uncontrolled cell proliferation disorder comprising: administering to an individual at least one disclosed compound; at least one disclosed pharmaceutical composition; and / or at least one product disclosed in a dosage and amount effective to treat the disorder in the subject. In a further aspect, there is provided a method for treating or preventing a neurodegenerative disorder comprising: administering to an individual at least one disclosed compound; at least one disclosed pharmaceutical composition; and / or at least one product disclosed in a dosage and amount effective to treat the disorder in the subject. Also provided is a method for treating a disorder in a mammal comprising the step of administering to the mammal at least one disclosed compound, composition or medicament.
[0180] The invention is directed to the use of the described chemical compositions for treating diseases or disorders in patients (preferably humans) where it could be predicted that LSD inhibition would have a therapeutic effect, such as disorders of uncontrolled cell proliferation. (e.g., cancers) and neurodegenerative disorders, such as Alzheimer's disease, Huntington's disease, and Parkinson's disease by administering one or more disclosed compounds or products.
The compounds disclosed in the present invention are useful in treating, preventing, ameliorating, controlling or reducing the risk of a variety of uncontrolled cell proliferation disorders. In one aspect, the uncontrolled cell proliferation disorder is associated with a histone demethylase dysfunction. In another aspect, histone demethylase dysfunction is dysregulation of LSD. In yet another aspect, histone demethylase dysfunction is dysregulation of LSD1. In yet another additional aspect, histone demethylase dysfunction is dysregulation of LSD2.
Also provided is a method of using a disclosed compound, composition or medicament. In one aspect, the method of use is directed to the treatment of a disorder. In a further aspect, the disclosed compounds may be used as individual agents or in combination with one or more other drugs in the treatment, prevention, control, amelioration or risk reduction of the above-mentioned diseases, disorders and conditions for which the compound or other drugs have utility, where the combination of drugs together are safer or more effective than either drug alone. The other drug (s) may be administered by one route and in an amount commonly used for that purpose, simultaneously or sequentially with a disclosed compound. When a disclosed compound is used simultaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compound is preferred. However, combination therapy may also be administered in overlapping schedules. It is also envisaged that the combination of one or more active ingredients and a disclosed compound may be more effective than either as a single agent.
Examples of disorders associated with a histone demethylase dysfunction include an uncontrolled cell proliferation disorder. In an additional aspect, the disorder of uncontrolled cell proliferation is cancer. Still in an additional aspect, cancer is a leukemia. In yet another additional aspect, cancer is a sarcoma. In yet another aspect, cancer is a solid tumor. Still in an additional aspect, cancer is a lymphoma.
Cancer is understood to refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Cancer may be multidrug resistant (MDR) or drug sensitive. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More particular examples of such cancers include breast cancer, prostate cancer, colon cancer, squamous cell cancer, small cell lung cancer, non-small cell lung cancer, gastrointestinal cancer, pancreatic cancer, cervical cancer. , ovarian cancer, peritoneal cancer, liver cancer, eg liver carcinoma, bladder cancer, colorectal cancer, endometrial carcinoma, kidney cancer, and thyroid cancer.
[0185] In many ways, other examples of cancers are basal cell carcinoma, biliary tract cancer, bone cancer; brain and CNS cancer; choriocarcinoma; connective tissue cancer; esophageal cancer; eye cancer; head and neck cancer; gastric cancer; intraepithelial neoplasia; laryngeal cancer; lymphoma, including Hodgkin's and non-Hodgkin's lymphoma; melanoma; myeloma; neuroblastoma; oral cavity cancer (eg, lips, tongue, mouth and pharynx); retinoblastoma; rhabdomyosarcoma; rectal cancer; respiratory system cancer; sarcoma; skin cancer; stomach cancer; testicular cancer; uterine cancer; urinary system cancer, as well as other carcinomas and sarcomas.
[0186] In another aspect, cancer is a hematological cancer. In an additional aspect, hematologic cancer is selected from acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia, chronic myelomonocytic leukemia (CMML). ), juvenile myelomonocytic leukemia (JMML), Hodgkin's lymphoma, non-Hodgkin's lymphoma, multiple myeloma, solitary myeloma, localized myeloma, and extramedullary myeloma. In yet another aspect, the cancer is selected from chronic lymphocytic leukemia, small lymphocytic lymphoma, non-Hodgkin B-cell lymphoma, and large B-cell lymphoma.
[0187] In an additional aspect, cancer is brain cancer. Still in an additional aspect, brain cancer is selected from a glioma, medulloblastoma, primitive neuroectodermal tumor (PNET), acoustic neurinoma, glioma, meningioma, pituitary adenoma, schwannoma, CNS lymphoma, primitive neuroectodermal tumor, craniopharyngioma, chordoma medulloblastoma, cerebral neuroblastoma, central neurocytoma, pineocytoma, pineoblastoma, atypical terdoid rhabdoid tumor, chondrosarcoma, chondroid, choroidal plexus carcinoma, craniopharyngioma, dysembrioplastic tumor neuroepithelial tumor, hematomalomatic tumor In yet a further aspect, the glioma is selected from ependymoma, astrocytoma, oligodendroglioma and oligoastrocytoma. In yet another additional aspect, the glioma is selected from juvenile pilocytic astrocytoma, subependymal giant cell astrocytoma, ganglioglioma, subependymoma, pleomorphic xanthoastrocytoma, anaplastic astrocytoma, glioblastoma multiforme, brain stem glioma, oligodendrogomalomatoma, anomalocytoma, anomalous infantile, subependymal giant cell astrocytoma, diffuse astrocytoma, mixed glioma, optic glioma, cerebral gliomatosis, multifocal gliomatous tumor, multicenter glioblastoma multiform tumor, paraganglioma and ganglioglioma.
[0188] In one aspect, cancer may be a cancer selected from blood, brain, genitourinary tract, gastrointestinal, colon, rectum, breast, kidney, lymphatic system, stomach, lung, pancreas and skin cancers. In an additional aspect, the cancer is selected from prostate cancer, glioblastoma multiforme, endometrial cancer, breast cancer and colon cancer. In an additional aspect, the cancer is selected from a breast, ovarian, prostate, head, neck and kidney cancer. In yet another aspect, the cancer is selected from cancers of blood, brain, genitourinary tract, gastrointestinal tract, colon, rectum, breast, liver, kidney, lymphatic system, stomach, lung, pancreas and skin. In yet another aspect, cancer is selected from lung and liver cancer. In yet another aspect, the cancer is selected from a breast, ovarian, testicular and prostate cancer. Still in an additional aspect, cancer is a breast cancer. Still in an additional aspect, cancer is an ovarian cancer. Still in an additional aspect, cancer is a prostate cancer. Still in an additional aspect, cancer is a testicular cancer.
[0189] In many respects, disorders associated with a histone demethylase dysfunction include neurodegenerative disorders. In another aspect, neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease and Huntington's disease.
In addition, the compounds are useful in a method for preventing, treating, controlling, ameliorating or reducing the risk of the diseases, disorders and conditions noted in this document. In addition, the compounds are useful in a method for preventing, treating, controlling, ameliorating or reducing the risk of the above-mentioned diseases, disorders and conditions in conjunction with other agents.
The present invention further relates to the administration of an LSD inhibitor to improve treatment outcomes in the context of uncontrolled cell proliferation disorders, including cancer. That is, in one aspect, the invention relates to a co-therapeutic method comprising the step of administering to a mammal an effective amount and dosage of at least one compound of the invention together with cancer therapy.
In a further aspect, administration improves treatment outcomes in the context of cancer therapy. Administration in the context of cancer therapy may be continuous or intermittent. Administration need not be concurrent with therapy and may occur before, during or after therapy. For example, cancer therapy may be provided 1, 2, 3, 4, 5, 6, or 7 days before or after administration of the compound. As an additional example, cancer therapy may be provided 1, 2, 3 or 4 weeks before or after administration of the compound. As yet another example, cognitive or behavioral therapy may be provided before or after administration within a time period of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 compound half-lives. administered.
In a further aspect, the disclosed compounds may be used in conjunction with one or more other drugs in the treatment, prevention, control, amelioration or reduction of the risk of diseases or conditions for which the disclosed compounds or other drugs have utility, where the drug combination is safer or more effective than any drug alone. Such other drug (s) may be administered by one route and in an amount commonly used for that purpose, simultaneously or sequentially with a compound of the present invention. When a compound of the present invention is used simultaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and a disclosed compound is preferred. However, combination therapy may also include therapies in which a disclosed compound and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in conjunction with one or more other active ingredients, the disclosed compounds and the other active ingredients may be used at lower doses than when each is used alone.
Accordingly, pharmaceutical compositions include those containing one or more other active ingredients in addition to a compound of the present invention.
The above combinations include combinations of a disclosed compound not only with another active compound, but also with two or more other active compounds. Likewise, the disclosed compounds may be used in conjunction with other drugs that are used in preventing, treating, controlling, ameliorating or reducing the risk of the diseases or conditions for which the disclosed compounds are useful. Such other drug (s) may be administered by one route and in an amount commonly used for that purpose, simultaneously or sequentially with a compound of the present invention. When a compound of the present invention is used simultaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to a disclosed compound is preferred. Accordingly, pharmaceutical compositions include those which also contain one or more other active ingredients in addition to a compound of the present invention.
The weight ratio of a disclosed compound to the second active ingredient may 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 a compound disclosed with the other agent will generally range from about 1000: 1 to about 1: 1000, preferably from about 200 : 1 to about 1: 200.
Combinations of a compound of the present invention and other active ingredients will generally also be within the above range, but in each case, an effective dose of each active ingredient should be used.
In such combinations, a disclosed compound and other active agents may be administered separately or together. In addition, administration of an element may occur before, at the same time as or after administration of other agent (s).
Thus, the subject compounds may be used alone or in combination with other agents that are known to be beneficial in the indications of the case or with other drugs that affect receptors or enzymes that increase efficacy, safety, convenience or reduce unwanted side effects or toxicity of the disclosed compounds. The compound of the matter and the other agent may be co-administered in concomitant therapies or in a fixed combination.
In one aspect, the compound may be employed in conjunction with anticancer therapeutic agents or other known therapeutic agents.
Under treatment conditions requiring inhibition or negative modulation of LSD protein activity, an appropriate dosage level will generally be about 0.01 to 1000 mg per kg body weight of the patient per day, and may be administered. in single or multiple doses. Preferably, the dosage level will be from about 0.1 to about 250 mg / kg per day; more preferably from about 0.5 to about 100 mg / kg per day. A suitable dosage level may be about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range, the dosage may be 0.05 to 0.5, 0.5 to 5, or 5 to 50 mg / kg per day. For oral administration, the compositions are preferably provided as tablets containing 1.0 to 1000 milligrams of active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200 , 250, 300, 400, 500, 600, 750, 800, 900 and 1000 milligrams of the active ingredient for symptomatic dose adjustment for the patient to be treated. The compounds may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosage regimen can be adjusted to provide the optimal therapeutic response. It will be understood, however, that the specific dose level and dosage frequency for any particular patient may vary, and will depend on a number of factors, including the activity of the specific compound used, the metabolic stability and the duration of action of that compound, age, body weight, general health status, gender, diet, mode and time of administration, excretion rate, drug combination, severity of the particular condition and the host undergoing therapy.
Thus, in one aspect, the invention relates to methods for negatively inhibiting or modulating LSD in at least one cell, comprising the step of contacting at least one cell with at least one compound of the invention in An effective amount modulates or activates the LSD activity response, for example, LSD1 or LSD2, in at least one cell. In another aspect, the cell is mammalian, e.g. human. In another aspect, the cell was isolated from an individual prior to the contact step. In another aspect, contact occurs through administration to an individual.
The. TREATMENT OF A PROLIFERATION DISORDER
Uncontrolled cell phone
In one aspect, the invention relates to a method for treating an uncontrolled cell proliferation disorder 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 uncontrolled cell proliferation disorder.
In yet a further aspect, the effective amount is a therapeutically effective amount. In yet another additional aspect, the effective amount is a prophylactically effective amount.
In a further aspect, the mammal is a human being. In a still further aspect, the method further comprises the step of identifying a mammal in need of treatment for an uncontrolled cell proliferation disorder. In another additional aspect, the mammal was diagnosed with a need for treatment of an uncontrolled cell proliferation disorder prior to the administration step.
In another aspect, the disorder of uncontrolled cell proliferation is associated with a histone demethylase dysfunction. In a further aspect, histone demethylase is a specific lysine histone demethylase. In a still further aspect, the specific lysine histone demethylase is LSD1. In a still further aspect, the specific lysine histone demethylase is LSD2.
In another aspect, the disorder of uncontrolled cell proliferation is a cancer. Still in an additional aspect, cancer is a leukemia. In yet another additional aspect, cancer is a sarcoma. In yet another aspect, cancer is a solid tumor. Still in an additional aspect, cancer is a lymphoma. Still in an additional aspect, the cancer is selected from chronic lymphocytic leukemia, small lymphocytic lymphoma, non-Hodgkin B-cell lymphoma and large B-cell lymphoma. In an additional aspect, the cancer is selected from cancers of the blood, brain, tract. genitourinary, gastrointestinal tract, colon, rectum, breast, liver, kidney, lymphatic system, stomach, lung, pancreas and skin. In yet another aspect, cancer is selected from lung and liver cancer. In yet another aspect, the cancer is selected from a breast, ovarian, testicular and prostate cancer. Still in an additional aspect, cancer is a breast cancer. Still in an additional aspect, cancer is an ovarian cancer. Still in an additional aspect, cancer is a prostate cancer. Still in an additional aspect, cancer is a testicular cancer.
B. HISTON DEMETILASE ACTIVITY DECREASE
In one aspect, the invention relates to a method for decreasing histone demethylase activity 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 The disclosed method of making a compound, or a pharmaceutically acceptable salt, hydrate, solvate or polymorph thereof, or a pharmaceutically acceptable hydrate, solvate, polymorph or salt thereof, thereby decreasing the activity of histone demethylase in the mammal.
In yet a further aspect, the effective amount is a therapeutically effective amount. In yet another additional aspect, the effective amount is a prophylactically effective amount.
In a further aspect, the mammal is a human being. In a still further aspect, the method further comprises the step of identifying a mammal in need of decreased histone demethylase activity. In a still further aspect, the mammal was diagnosed with a need to decrease histone demethylase activity prior to the administration step.
In a further aspect, histone demethylase is a specific lysine histone demethylase. In a still further aspect, the specific lysine histone demethylase is LSD1. In a still further aspect, the specific lysine histone demethylase is LSD2.
In an additional aspect, the need to decrease histone demethylase activity is associated with a histone demethylase dysfunction. In yet a further aspect, histone demethylase dysfunction is associated with an uncontrolled cell proliferation disorder. In a still further aspect, the method further comprises the step of identifying a mammal in need of treatment for an uncontrolled cell proliferation disorder. In another additional aspect, the mammal was diagnosed with a need for treatment of an uncontrolled cell proliferation disorder prior to the administration step.
In yet another aspect, the disorder of uncontrolled cell proliferation is a cancer. Still in an additional aspect, cancer is a leukemia. In yet another additional aspect, cancer is a sarcoma. In yet another aspect, cancer is a solid tumor. Still in an additional aspect, cancer is a lymphoma. Still in an additional aspect, the cancer is selected from chronic lymphocytic leukemia, small lymphocytic lymphoma, non-Hodgkin B-cell lymphoma and large B-cell lymphoma. In an additional aspect, cancer is selected from blood, brain, genitourinary tract, gastrointestinal tract, colon, rectum, breast, liver, kidney, lymphatic system, stomach, lung, pancreas and skin. In yet another aspect, cancer is selected from lung and liver cancer. In yet another aspect, the cancer is selected from a breast, ovarian, testicular and prostate cancer. Still in an additional aspect, cancer is a breast cancer. Still in an additional aspect, cancer is an ovarian cancer. Still in an additional aspect, cancer is a prostate cancer. Still in an additional aspect, cancer is a testicular cancer.
ç. HISTON DEMETILASE ACTIVITY DECREASE
IN CELLS
In one aspect, the invention relates to a method for decreasing histone demethylase activity in at least one cell, the method comprising the step of contacting at least one cell with an effective amount of at least one cell. disclosed compound or a product of a disclosed method of making a compound, or a pharmaceutically acceptable salt, hydrate, solvate or polymorph thereof, or a pharmaceutically acceptable hydrate, solvate, polymorph or salt thereof, thereby decreasing histone demethylase activity in the cell.
In yet a further aspect, the effective amount is a therapeutically effective amount. In yet another additional aspect, the effective amount is a prophylactically effective amount.
In a further aspect, the cell is mammalian. In yet another aspect, the cell is of a human being. In yet a further aspect, contact occurs through administration to a mammal. In another aspect, the method further comprises the step of identifying the mammal as having a need to decrease histone demethylase activity in a cell. In yet another aspect, the mammal was diagnosed with a need to decrease histone demethylase activity prior to the administration step.
In another aspect, histone demethylase is a specific lysine histone demethylase. In a still further aspect, the specific lysine histone demethylase is LSD1. In a still further aspect, the specific lysine histone demethylase is LSD2.
[0217] In another aspect, the need to decrease histone demethylase activity in a cell is associated with a lack of cellular control.
In yet another aspect, the disorder of uncontrolled cell proliferation is a cancer. Still in an additional aspect, cancer is a leukemia. In yet another additional aspect, cancer is a sarcoma. In yet another aspect, cancer is a solid tumor. Still in an additional aspect, cancer is a lymphoma. Still in an additional aspect, the cancer is selected from chronic lymphocytic leukemia, small lymphocytic lymphoma, non-Hodgkin B-cell lymphoma and large B-cell lymphoma. In an additional aspect, the cancer is selected from cancers of the blood, brain, tract. genitourinary, gastrointestinal tract, colon, rectum, breast, liver, kidney, lymphatic system, stomach, lung, pancreas and skin. In yet another aspect, cancer is selected from lung and liver cancer. In yet another aspect, the cancer is selected from a breast, ovarian, testicular and prostate cancer. Still in an additional aspect, cancer is a breast cancer. Still in an additional aspect, cancer is an ovarian cancer. Still in an additional aspect, cancer is a prostate cancer. Still in an additional aspect, cancer is a testicular cancer.
2. MANUFACTURE OF A MEDICINAL PRODUCT
In one aspect, the invention relates to a method for the manufacture of a medicament for inhibiting histone demethylase activity in a mammal which comprises combining a therapeutically effective amount of a disclosed compound or product of a disclosed method. a pharmaceutically acceptable carrier or diluent.
F. EXPERIMENTAL
The following examples are provided to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed in the present invention are made and evaluated, and are intended to be purely exemplary of the invention, and not to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers (eg quantities, temperature, etc.) but some errors and deviations should be taken into account. Unless otherwise indicated, parts are parts by weight, temperature is in ° C or room temperature, and pressure is at or near atmospheric pressure.
Several methods for preparing the compounds of this invention are illustrated in the following examples. The necessary starting materials and intermediates are in some cases commercially available, or may be prepared according to literature procedures or as illustrated herein.
The following exemplary compounds of the invention were synthesized. Examples are provided herein to illustrate the invention and should not be construed as limiting the invention in any way. Examples are usually portrayed as a free base according to the IUPAC naming convention. However, some of the examples were obtained or isolated in salt form.
As indicated, some of the examples were obtained as racemic mixtures of one or more enantiomers or diastereoisomers. The compounds may be separated by one of ordinary skill in the art to isolate individual enantiomers. Separation may be accomplished by coupling a racemic mixture of the compounds with an enantiomerically pure compound to form a diastereoisomeric mixture, followed by separation of the individual diastereoisomers by standard methods such as fractional crystallization or chromatography. A racemic or diastereoisomeric mixture of the compounds may also be separated directly by chromatographic methods using chiral stationary phases.
1. CHEMICAL MATERIALS AND GENERAL METHODS
All analytical grade or anhydrous reagents were purchased from commercial sources and were used without further purification. The solvents were analytical or anhydrous grade (Sigma-Aldrich). Special chemicals and building blocks obtained from various suppliers had the highest purity offered (always> 95%).
NMR spectroscopy was performed on a Varian Unity 400 instrument with a 5 mm wide band probe and using standard pulse sequences. Chemical displacements (δ) are reported in parts per million (ppm) in the low field of solvent references. Coupling constants (J values) are expressed in Hz.
Mass spectrometry was performed on a LCMS Finnigan LCQ Duo ion trap electrospray ionization (ESI) mass spectrometer. All samples were analyzed by positive ESI-MS and mass-charge ratio (m / z). ) of the protonated molecular ion is reported.
Microwave assisted reactions were performed on a Biotage 2.5 primer at various potencies.
Hydrogenation reactions were performed in a standard Parr hydrogenation apparatus.
[0228] Reactions were monitored by HPLC or TLC. When monitored by TLC, reactions were analyzed on 200 µm Baker flexible reinforced silica gel plates containing a fluorescent indicator. Preparative TLC was performed on 20 cm x 20 cm Uniplates Analtech plates coated with a 1000 or 2000 µm silica gel layer containing a fluorescent indicator (UV 254).
Elution mixtures are reported as v: v. Spot visualization was achieved using UV light.
Flash chromatography was performed on a Teledyne Isco CombiFlash RF 200 using C-18 reversed phase or appropriately sized Redisep Rf Gold standard phase silica columns. The crude compounds were adsorbed on silica gel, 70 to 230 mesh 40 Å (for normal phase) or Celite 503 (for reverse phase) and loaded into solid cartridges. Elution mixtures are reported as v: v.
2. MOLECULAR MODELING AND VIRTUAL SCREENING METHODS
[0230] All computational studies used PDB ID 2Z5U for the LSD1 structural coordinates. The ICM, Glide and GOLD virtual coupling methods programs have been implemented. The protein structure was prepared by 3D protonation, water molecule elimination and energy minimization using the ICM force field and distance-dependent dielectric potential with an RMS gradient of 0.1; heavy atoms in the protein were kept fixed, and histidine residues were considered neutral. Virtual screening calculations used standard parameters (unless explicitly specified otherwise) with ICM and Glide scores being scoring functions, respectively. In both cases, FAD was defined as the ligand and an active site region was defined by a 12 Â radius sphere around the FAD bound in the LSD1 complex.
Confirmation of the precision and efficiency of the coupling protocol applied used the FAD dinucleotide adenine cofactor fragment and the flavin fragment, and known LSD1 inhibitors (decoy assembly) as positive controls. Two separate coupling analyzes were performed with the ICM and Glide coupling program; The GOLD coupling program was used for a new classification.
[0232] The compound database was prepared using Ligprep 2.1.23 (Schrodinger, LLC., New York, New York). Two VS cycles including HTVS and standard precision coupling (SP) were adopted. The 10,000 major compounds classified by Glide were stored and subjected to additional coupling experiments using ICM coupling. The final set of 2000 hits was selected based on ICM scores and the individual compounds were visually inspected for coupling representations and interactions between ligands and LSD1. The GOLD consensus scoring functions were additionally used to retell these 2000 hits selected by Glide and ICM. Finally, 121 compounds were purchased (if available) or synthesized for LSD1 inhibition studies.
3. MD SIMULATION METHODS
All simulations were performed using the ff99SB AMBER force field (Hornak, V., et al. Proteins 2006, 65 (3), 712-25) for LSD1, the Amber general force field ("gaff"; see Wang, J., et al. J. Comput Chem 2004, 25 (9), 1157-74) for compound 12, and the TIP3P model (Journal of Chemical Physics 1982, (77), 4156-4163) for water were used. Simulations approximate long-range electrostatic interactions using the Ewald particle-mesh (PME) method procedure (Essmann, U., et al.
Journal of Chemical Physics 1995, (103), 8577-8593; Darden, T., et al. Journal of Chemical Physics 1993, (98), 10089-1092). Using LeaP, the binding modes generated from the ICM coupling on the LSD1 complex were solvated to neutral charge and the complexes were first minimized with PMEMD (Case, D. A., et al. AMBER11, San Francisco, 2010). After minimization, 200 ps of unrestricted dynamic molecular simulation using an unbound cutoff limit of 9Â ° was performed for both coupling modes with a periodic constant pressure limit maintaining 1 atm of pressure and isotropic position sizing over time. 2 ps relaxation SHAKE was used to constrain hydrogen bonding, and Langevin dynamics were used to regulate temperature (Case, DA, et al. AMBER11, San Francisco, 2010), keeping at 300 K. Bond-free relative energies for comparisons between Both binding modes were predicted using MMPBSA.py9 with 100 photos at 1-ps intervals, starting at lps or 10lps in the trajectory.
4. VIRTUAL SCAN RESULTS
[0234] The first crystalline structures of LSD1 elucidating critical structural features were later presented by Stavropoulos et al. (Nat Struct Mol Biol 2006, 13 (7): 626-32; Protein Data Bank or PDB ID 2H94; see http://www.wwpdb.org/), Yang et al. (Mol Cell 2006, 23 (3) , 377-87; PDB ID 2IW5), and Chen et al. (Proc Natl Acad Sci USA 2006, 103 (38), 13956-61; PDB ID 2HKO). These 2.9 Â, 2.57 Â, and 2.8 Â structures, respectively, show a highly negatively charged substrate binding cavity spacious enough to accommodate the N-terminal tail of histone H3. In addition, an N-terminal SWIRM domain and an insertion into the catalytic core domain, called the Tower Domain, were defined as structural portions required for enzymatic activity and interactions with cofactors such as CoREST. For the studies described herein, the structure, PDB ID 2Z5U, was used with tranilcipromin-bound LSD1 inhibitor for computational studies, including virtual scanning, coupling, and molecular dynamics (Mimasu, S., et al. Biochem Biophys Res Commun 2008, 366 (1), 15-22). In order to evaluate the sternal chemical space of tranylcypromin and polyamine derivatives, HTVS was used with a company library. The library was cured from publicly available vendor libraries, totaling about 13 million compounds, using internally developed custom filters. Compounds were filtered based on the Lipinski rule of five, with exceptions occurring in only 62,000 compounds. In addition, structurally redundant compounds were removed so that the resulting library contained a diverse yet manageable set of about 2 million compounds. Prior to screening, the compounds were prepared using the Schrodinger suite's LigPrep module as well as ICM's integrated preparation of three-dimensional (3D) ligands so that physiologically relevant protonation states were used.
[0235] The prepared ligands were then coupled against three different sites on LSD1; the FAD site located in the amine oxidase domain and the adenine dinucleotide and flavin fragments of this "pocket". The coupling protocols used by ICM and Glide were performed with FAD, dinucleotide adenine, and flavin fragments, and with known LSD1 inhibitors to verify accuracy. In addition to the coupling algorithm classifications, visual inspection of the coupling results was used to assess proper coupling position, posture, and orientation. Taken together, the ICM and Glide scoring functions were able to correctly identify known inhibitors contained in the top 2% of the decoy set used. GOLD was used to repackage and the GOLD adjustment function produced similar enrichments.
[0236] A virtual scan was set against the LSD1 FAD binding pocket using the established coupling protocol established and the database containing 2 million compounds. The 10,000 major compounds were selected from both the ICM and Glide scoring functions for further analysis. Some identical compounds were scored similarly between the two algorithms; This redundancy has been filtered out. In addition, visual inspection was performed to filter out similar compounds and to increase the diversity of the final selection. Visual analysis also identified the most important interactions in the LSD1 ADF pocket. These include hydrogen bonding with Ser289, Arg310 and Arg316, van der Waals interactions with Val590 and Leu625 and π interactions with Trp756. In addition, hydroxyl compounds and hydrophobic electron removal groups appeared to show greater enrichment in the initial coupling results. The LSD1 FAD binding pocket is a narrow, deep slit inside the protein and is surrounded by hydrophobic amino acid residues. Thus, the hydrophobic character of the compounds may play an important role in the random pathway of the compound at the active site.
Based on the selection criteria discussed above, 121 structurally distinct compounds were obtained and sent for biochemical screening with LSD1. The biochemical assay, as described in the experimental section, measures H2O2 produced from oxidative demethylation of a peptide substrate. Of the 121 compounds, a series of related compounds that showed potent activity in the biochemical assay were identified. Coupling scores, ratings, and results of the corresponding biochemical assay for the series are shown in Tables 1, 2, and 6 to 9.
Of the ten active compounds in Table 1 (and associated tables, which provide biochemical and cellular data, Tables 6, 8, and 9), which were discovered using virtual scanning methods, for example, compounds 1, 2, 4, and 5 showed similar binding modes at the LSD1 FAD binding site. In addition, coupling scores for compounds 1, 2, 4 and 5 correlated well with observed biochemical activity. These results suggested that improved pocket-directed adenine dinucleotide inhibitors in the LSD1 amine oxidase domain were accessible.
Glide scores are predictive and correlate with compounds with aryl p-OH or m-Cl substitutions (compounds 1 and 5). It is evident from these studies that hydrophobic electron removal groups, such as -Cl, are tolerated, while small alkyl substituents such as methyl (eg, compound 8) or fused bicyclic substituents containing compound 10 have less activity. The introduction of any donor groups, particularly the -OCH3 functional group at the 2nd position, loses activity due to the lack of H-binding interactions with Gly314 (e.g., compound 6). The lack of biochemical activity of compound 6 was highly predictive of coupling scores, where ICM and Glide provided energies of -18.39 and -6.63 kcal / mol, respectively. In subsequent coupling analysis, series of additional benzohydrazine compounds were identified, with compounds containing 4-substituted sulfone -C methyl or aryl hydrazine, as exemplified by virtual matching compound 9, which exhibited potent LSD1 inhibiting activity with an IC 50 of 19 nM. The low coupling score of compound 9 is mainly due to the shift in position of the 2-OH aryl ring. Compound 9, with a sulfone / morpholine substituent, was chosen as a structure for further optimization due in part to its chemical stability.
The mode of binding of compound 12 to the sulfone / morpholine is depicted with the predicted coupling representation from the ICM in Figure 1. In this model, the phenolic group fits well in the pocket comprising residues Ser289, Gly314 and Arg316. . The central carbonyl group appears to be involved in strong H-binding interactions with the Arg310 amino group and morpholine oxygen exhibits H-binding interactions with Val590.
These sets of hydrogen bonding interactions were also observed with the Glide and GOLD coupling experiments. Further experiments showed the morpholine substituted aryl ring participating in n-n interactions with the Trp756 residue, while morpholine oxygen was retained at the H bond with Val590.
Chemical optimization also focused on the design of compounds containing heteroaryl rings on either side of compound 12. Computer models using these results generated a variety of chemically plausible structures, of which a substituted pyridine was identified as an appropriate moiety capable. interact with the Ser289, Gly314 and Arg316, the surrounding residues and with the ideal properties. One representative is compound 24, which showed potent LSD1 activity (28 nM) and also exhibited a binding mode similar to that of compound 12 (see Figure 2).
Many of the representative compounds contain a C-alkyl hydrazine to increase the metabolic stability of the series. However, a larger group, such as the ethyl group of compound 21, is not well accommodated by the binding pocket as illustrated by the different biochemical activities of compounds 12 and 21. The replacement of aryl with methylsulfone (compound 25) is substituted One morpholine ring (compound 12) increased biochemical efficacy by about one order of magnitude compared to compound 11. The addition of just one morpholine ring maintains some biochemical activity, as illustrated by compound 23. Sulfonmorpholine replacement Sulfono-N-dimethyl also maintained biochemical activity, as illustrated by compound 18. In addition, it was found that substitution of the 2-OH group with a chlorine was not well housed and a significant drop in activity was demonstrated between Compounds 12 and 16. Results with compound 24 suggest that the use of a substituted pyridine is accommodated by the enzyme, but several other substitutions and heterocycles generally resulted in a drop in biochemical activity, as illustrated in compounds 13, 14, 15, 17, 19, 20 and 22.
Many of the representative compounds in Table 2 contained a C-alkylhydrazine to increase the metabolic stability of the series. However, a larger group, for example, The ethyl group of compound 21, is less well accommodated by the binding pocket, as illustrated by the different biochemical activities of compounds 12 and 21. Aryl substitution with methylsulfone (e.g. 25) and substituted with a morpholine ring (compound 12) increased biochemical efficacy by about one order of magnitude compared to compound 11. Addition of a heterocycle, for example a morpholine ring, maintains biochemical activity, As shown by compound 23. Substitution of sulfonomorpholine with sulfono-N-dimethyl also maintained biochemical activity, as illustrated by compound 18. In addition, substitution of the 2-OH group with a chlorine was found not to be successful. housed, with a significant drop in activity between compounds 12 and 16.
As discussed above, compound 24 suggests that the use of a substituted pyridine is adapted by the enzyme. Further analysis suggests that the hydroxyl of compound 12 is associated with increased biochemical activity, for example, when this substituent group is replaced with a chlorine (compound 16), the activity decreased. TABLE 1
5. RESULTS OF SIMULATION OF MOLECULAR DYNAMICS
Molecular dynamics ("MD") simulations were performed using the two different coupling representations of compound 12 to determine if there was a preference for one coupling representation over another. These data can better inform which interactions play a role in the results obtained with the synthesized compounds. Coupling results show the highest ranked representation of compound 12 bound in the dinucleotide binding pocket by direct H-binding interactions with Ser289 or Arg316 through its hydroxyl moiety (binding mode 1, see figure 3 and table 3). However, there is another favorably punctuated representation with the morpholine ring of compound 12 interacting with Ser289 and Arg316 (binding mode 2, see Figure 3 and Table 3).
[0245] MD using the AMBER program set was used to evaluate the binding energy for both predicted binding modes. Simulations for bonding mode 1 showed the interactions of π-conjugated electrons between compound 12 and Arg 316, as well as the potential for hydrogen bonding between hydroxyl and Ser289. Analysis of binding mode 2 showed potential n-n interactions between compound 12 and Trp756, with more favorable hydrogen bonds with Arg310 and Arg316. In addition, bonding mode 1 is predicted to have hydrogen bonding with Val590, while bonding mode two has van der Waals interactions involving the chlorine group. MMPBSA analysis of the final 100 ps simulation showed that it is predicted that binding mode 2 has a free binding energy of -40.8 Kcal / mol, which is approximately 20 Kcal / mol more favorable than -21.0 for binding mode 1. The first 100 ps of simulation probably partly reflects the equilibrium of the complex, so the calculated free binding energies are not as favorable. This finding contradicts the classifications of binding representations during the coupling process. This difference may arise from differences in protein structure during coupling and MD, with a rigid structure used to increase coupling protocol speed and a flexible structure used for MD. TABLE 3. MD Compound No. 12 Compound No. 12 (Bonding Mode 1) (Bonding Mode 2) 1 at 100ps: -20.2154 -32.9117 Bonding AG (Kcal / mol) 101-200ps: -21 0.263 -40.8046 Binding Ag (Kcal / mol) 150-200ps: 0.394 1.560 RMSD of Binder (Δ) 6. PREPARATION OF (E) -N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE ) BENZOHYDRAZED.
1- (5-chloro-2-hydroxyphenyl) ethanone (100 mg, 0.586 mmol) and benzohydrazide (80 mg, 0.586 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and in Then, the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. After cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound (90 mg) as a solid. 1H-NMR (400 MHz, DMSO-d6): δ 7.95 (m, 2H), 7.67-7.62 (m, 2H), 7.56 (m, 2H), 7.35 (dd, 1H , J = 2.4 & 8.8 Hz), 6.95 (d, 1H, J = 8.4 Hz), 3.35 (s, 3H). ESI-MS: 289.0 [M + H] +. 7. PREPARATION OF (E) -N (1- (2,6 — DI — HYDROXYPHENYL) ETHYLIDENE) BENZOHIDRAZID.
1- (2,6-dihydroxyphenyl) ethanone (100 mg, 0.657 mmol) and benzohydrazide (89 mg, 0.657 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as catalyst and then , the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. After cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound (100 mg) as a solid. 1 H NMR (400 MHz, CD 3 OD): δ 7.59 (m, 2H), 7.49 (m, 1H), 7.39 (m, 2H), 7.11 (t, 1H, J = 8.0 Hz), 6.45 (m, 2H), 2.35 (s, 3H). ESI-MS: 271.1 [M + H] +. 8. PREPARATION OF 3- (MORPHOLINOSULPHONYL) BENZOIC ACID.
3- (Chlorosulfonyl) benzoic acid (250 mg, 1.133 mmol) was added to morpholine (99 mg, 1.133 mmol) in the presence of potassium carbonate (313 mg, 2.266 mmol) in THF (5 mL) at room temperature. , and the reaction mixture was stirred for 12 h at room temperature. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (3% CH 3 OH / CH 2 Cl 2) yielding the title compound (160 mg) as a solid. 1 H NMR (400 MHz, CD 3 OD): δ 8.34 (m, 1H), 8.32 (d, 1H, J = 8.0 Hz), 7.99 (m, 1H), 7.76 (t, 1H, J = 8.0 Hz), 3.70 (m, 4H), 2.98 (m, 4H). ESI-MS: 272.0 [M + H] +.
9. PREPARATION OF 3- (MORPHOLINOSULPHONYL) METZO BENZOATE.
3- (Morpholinosulfonyl) benzoic acid (100 mg, 0.369 mmol) was refluxed overnight in methanol in the presence of catalytic concentrated H2SO4 at 65 ° C. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography to afford the title compound as an off-white solid (60 mg). NMR (400 MHz, CDCl 3): δ 8.38 (t, 1H, J = 1.6 Hz), 8.27 (m, 1H), 7.92 (m, 1H), 7.64 (t, 1H , J = 8.0 Hz), 3.95 (s, 3H), 3.73 (m, 4H), 3.00 (m, 4H). ESI-MS: 286.1 [M + H] +. 10. PREPARATION OF 3- (MORPHOLINOSULPHONYL) BENZOHIDRAZID.
Methyl 3- (morpholinosulfonyl) benzoate (120 mg, 0.421 mmol) was added to hydrazine (17.52 mg, 0.547 mmol) in methanol and refluxed for 12 hours at 65 ° C. The reaction was monitored by TLC. After completion of the reaction and cooling of the reaction mixture, the solvent was removed under vacuum and then the compound was purified by column chromatography to afford the title compound as an off-white solid (90 mg). 1H-NMR (400 MHz, CDCl3): δ 8.16 (m, 1H), 8.12 (m, 1H), 8.04 (m, 1H), 7.85 (m, 1H), 7.63 ( t, 1H, J = 8.0 Hz), 4.19 (m, 2H), 3.71 (m, 4H), 2.97 (m, 4H). ESI-MS: 286.1 [M + H] +. 11. PREPARATION OF (E) -Ν '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
1- (5-chloro-2-hydroxyphenyl) ethanone (20 mg, 0.117 mmol) and 3- (morpholinosulfonyl) benzohydrazide (33.5 mg, 0.117 mmol) were dissolved in methanol (4 mL) in the presence of acid acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. After cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound (16 mg) as a solid. 1 H NMR (400 MHz, CD 3 OD): δ 8.26 (m, 1H), 8.17 (d, 1H, J = 8.0 Hz), 7.92 (d, 1H, J = 8.0 Hz), 7.72 (t, 1H, J = 8.0 Hz), 7.48 (d, 1H, J = 2.0 Hz), 7.22 (m, 1H), 6.91 (d, 1H, J = 8.8 Hz), 3.72 (m, 4H), 3.01 (m, 4H), 2.43 (s, 3H). ESI-MS: 438.1 [M + H] +. 12 PREPARATION OF (E) -N '- (1- (3-CHLORUS-2-FLUORPHENYL) ETHYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
1- (3-chloro-2-fluorophenyl) ethanone (20 mg, 0.116 mmol) and 3- (morpholinosulfonyl) benzohydrazide (33.1 mg, 0.116 mmol) were dissolved in methanol (4 mL) in the presence of acid acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. After cooling, the solvent was removed by vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound (22 mg) as a solid. 1 H NMR (400 MHz, CDCl 3): δ 9.43 (s, 1H), 8.37 (m, 1H), 8.16 (m, 1H), 7.87 (d, 1H, J = 7.2 Hz), 7.65 (m, 1H), 7.41 (m, 1H), 7.10 (t, 1H, J = 8.0 Hz), 3.71 (m, 4H), 2.95 ( m, 4H), 2.38 (s, 3H). ESI-MS: 440.1 [M + H] +. 13. PREPARATION OF (E) -N '- (1- (2-CHLOROPYRIDIN — 4 — IL) ETHYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
1- (2-chloropyridin-4-yl) ethanone (20 mg, 0.129 mmol) and 3- (morpholinosulfonyl) benzohydrazide (36.7 mg, 0.129 mmol) were dissolved in methanol (4 mL) in the presence of acid acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. After cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography, yielding the title compound in 60% yield. 1 H NMR (400 MHz, CDCl 3): δ 9.43 (m, 1 H), 8.39 (m, 2 H), 8.15 (d, 1H, J = 8.0 Hz), 7.93 (d, 1H, J = 7.6 Hz), 7.70 (t, 1H, J = 7.6 Hz), 7.52 (m, 1H), 3.73 (m, 4H), 3.02 (m, 4H), 2.35 (s, 3H). ESI-MS: 423.1 [M + H] +. 14 PREPARATION OF (E) -N (1- (2,5 — DICHLOROPHENYL) ETHYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
1- (2,5-dichlorophenyl) ethanone (20 mg, 0.106 mmol) and 3- (morpholinosulfonyl) benzohydrazide (30.2 mg, 0.106 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. After cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography, yielding the title compound in 10 mg yield. 4H NMR (400 MHz, CDCl3): δ 8.29 (m, 1H), 8.09 (m, 1H), 7.81 (m, 1H), 7.57 (m, 1H), 7.40 ( m, 1H), 7.26 (m, 2H), 3.52 (m, 4H), 2.91 (m, 4H), 2.28 (s, 3H). ESI-MS: 456.1 [M + H] +. 15 PREPARATION OF 4-HYDRAZINYL-3- (MORPHOLINOSULPHONYL) METHYL BENZOATE.
Methyl 4-fluoro-3- (morpholinosulfonyl) benzoate (30 mg, 0.099 mmol) was added to hydrazine (4.44 mg, 0.138 mmol) in methanol (8 mL) and refluxed for 5 h at 65 °. Ç. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (20 mg). NMR (400 MHz, CD 3 OD): δ 8.15 (d, 1H, J = 2.0 Hz), 8.03 (dd, 1H, J = 2.4 & 9.2 Hz), 7.48 (d 1H, J = 9.2 Hz), 3.86 (s, 3H), 3.67 (m, 4H), 3.04 (m, 4H). ESI-MS: 316.1 [M + H] +. 16. PREPARATION OF 4-FLUOR-3- (MORPHOLINOSULPHONYL) METHYL BENZOATE.
4-Fluoro-3- (morpholinosulfonyl) benzoic acid (50 mg, 0.173 mmol) was refluxed overnight in the presence of concentrated sulfuric acid (1.117 mg, 8.64 pmol) in methanol ( 8 mL) at 70 ° C. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (20 mg). 1 H NMR (400 MHz, CD 3 OD): δ 8.42 (dd, 1H, <7 = 2.0 & 6.4 Hz), 8.33 (m, 1H), 7.49 (t, 1H, J = 8.8 Hz), 3.94 (s, 3H), 3.71 (m, 4H), 3.16 (m, 4H). 17 PREPARATION OF 3-BROMO-4 — METHYL CHLOROBENZOATE.
3-Bromo-4-chlorobenzoic acid (200 mg, 0.849 mmol) was refluxed in the presence of concentrated sulfuric acid (5.49 mg, 0.042 mmol) in methanol (10 mL) at 70 ° C overnight To the other. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography to yield the title compound (130 mg). 1 H NMR (400 MHz, CDCl 3): δ 8.29 (d, 1H, J = 2.0 Hz), 7.91 (dd, 1H, J = 2.0 & 8.4 Hz), 7.52 ( d, 1H, J = 8.4 Hz), 3.92 (s, 3H). ESI-MS: 250.9 [M + H] +.
18. PREPARATION OF 3- (N, N — DIMETHYLSULFAMOIL) METHYL BENZOATE.
3- (N, N-Dimethylsulfamoyl) benzoic acid (200 mg, 0.872 mmol) was refluxed in the presence of concentrated sulfuric acid (5.64 mg, 0.044 mmol) in methanol (10 mL) at 70 ° C . The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography to yield the title compound (125 mg). 1 H NMR (400 MHz, CDCl 3): δ 8.42 (s, 1H), 8.27 (d, 1H, J = 8.0 Hz), 7.97 (d, 1H, J = 7.2 Hz) 7.65 (t, 1H, J = 8.0 Hz), 3.96 (s, 3H), 2.74 (s, 6H). ESI-MS: 244.0 [M + H] +. 19. PREPARATION OF 3 — BROMO — 4 — CHLOROBENZOHIDRAZID.
Methyl 3-bromo-4-chlorobenzoate (120 mg, 0.481 mmol) was added to hydrazine (23.12 mg, 0.721 mmol) in methanol (8 mL) and refluxed for 12 hours at 70 ° C. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography to yield the title compound (30 mg). 1 H NMR (400 MHz, CDCl 3): δ 8.02 (d, 1H, J = 1.6 Hz), 7.60 (dd, 1H, J = 2.0 & 8.0 Hz), 7.52 ( d, 1H, J = 8.0 Hz). ESI-MS: 250.9 [M + H] +. 20 PREPARATION OF 3- (HYDRAZINOCARBONYL) —N, N— DIMETHYLBenzenesulfonamide.
Methyl 3- (N, N-dimethylsulfamoyl) benzoate (150 mg, 0.617 mmol) was added to hydrazine (29.6 mg, 0.925 mmol) in methanol (10 mL) and refluxed for 8 h at 65 °. Ç. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (60 mg). 1H-NMR (400 MHz, CDCl3): δ 8.11 (s, 1H), 8.01 (d, 1H, J = 8.4 Hz), 7.92 (d, 1H, J = 8.0 Hz) , 7.65 (t, 1H, J = 8.0 Hz), 2.73 (s, 6H). ESI-MS: 244.0 [M + H] +. 21. PREPARATION OF (E) -3-BROMO-4-CHLOR-N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) BENZOHYDRAZIDE.
3-bromo-4-chlorobenzohydrazide (30 mg, 0.120 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (20.51 mg, 0.120 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound (15 mg). 1 H NMR (400 MHz, acetone-cfe): δ 8.30 (s, 1H), 7.98 (d, 1H, J = 8.4 Hz), 7.73 (d, 1H, J = 8.4 Hz), 7.61 (d, 1H, J = 2.4 Hz), 7.29 (dd, 1H, J = 2.4 & 8.4 Hz), 6.93 (d, 1H, J = 8 , 8 Hz), 2.55 (s, 3H). ESI-MS: 402.9 [M + H] +. 22 PREPARATION OF (E) -3- (2- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) HYDRAZINOCARBONYL) —N, N — Dimethylbenzenesulfonamide.
3- (hydrazinocarbonyl) -N, N-dimethylbenzenesulfonamide (50 mg, 0.206 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (35.1 mg, 0.206 mmol) were dissolved in methanol ( 4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound as a solid (15 mg). 1H-NMR (400 MHz, acetone-c6): δ 8.29 (m, 2H), 8.01 (d, 1H, J = 8.4 Hz), 7.83 (t, 1H, J = 8.4 Hz), 7.62 (d, 1H, J = 2.4 Hz), 7.32 (dd, 1H, J = 2.4 & 8.8 Hz), 6.96 (d, 1H, J = 8 , 8 Hz), 2.73 (s, 6H), 2.58 (s, 3H). ESI-MS: 396.0 [M + H] +. 23. PREPARATION OF 5-BROMO-6 — CHLORONICOTINOHYDRAZED.
Methyl 5-bromo-6-chloronicotinate (100 mg, 0.399 mmol) was added to hydrazine (19.19 mg, 0.599 mmol) in methanol (8 mL) and heated overnight at 70 ° C. . The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (20 mg). 4 H NMR (400 MHz, CD 3 OD): δ 8.33 (d, 1H, J = 2.4 Hz), 8.01 (d, 1H, J = 2.4 Hz). 24 PREPARATION OF (E) -5-BROMO-6-CHLOR-N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) NICOTINOHYDRAZIDE.
5-Bromo-6-chloronicotinohydrazide (15 mg, 0.060 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (10.22 mg, 0.060 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound as a solid (8 mg). 7Η NMR (400 MHz, DMSO-d 6): δ 8.39 (d, 1H, J = 2.4 Hz), 8.28 (s, 1H), 7.63 (d, 1H, J = 2.4 Hz), 7.32 (dd, 1H, J = 2.4 & 8.8 Hz), 7.06 (d, 1H, J = 6.8 Hz), 6.92 (d, 1H, J = 9 , 2 Hz), 6.81 (d, 1H, J = 6.8 Hz), 2.47 (s, 3H). ESI-MS: 404.0 [M + H] +. 25. PREPARATION OF METHYL 5-CHLORONICOTINATE.
5-Chloronicotinic acid (200 mg, 1.269 mmol) was refluxed overnight in the presence of concentrated sulfuric acid (8.20 mg, 0.063 mmol) in methanol (10 mL) at 70 ° C . The reaction was monitored by TLC. Upon completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (120 mg). 1H-NMR (400 MHz, CDCl3): δ 9.07 (d, 1H, J = 1.6 Hz), 8.72 (d, 1H, J = 2.0 Hz), 8.26 (m, 1H) 3.95 (s, 1H).
26. PREPARATION OF METHYL 5-CHLORONICOTINATE
5-Chloronicotinic acid (200 mg, 1.269 mmol) was refluxed overnight in the presence of concentrated sulfuric acid (8.20 mg, 0.063 mmol) in methanol (10 mL) at 70 ° C . The reaction was monitored by TLC. Upon completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (120 mg). 1 H NMR (400 MHz, CDCl 3): δ 9.07 (d, 1H, δ = 1.6 Hz), 8.72 (d, 1H, J = 2.0 Hz), 8.26 (m, 1H ), 3.95 (s, 1H). 27 PREPARATION OF 5-CHLORONICOTINOHYDRAZED.
Hydrazine (17.93 mg, 0.560 mmol) was added to methyl 5-chloronicotinate (80 mg, 0.466 mmol) in methanol (8 mL) and heated overnight at 70 ° C. The reaction was monitored by TLC. Upon completion of the reaction, the solvent was removed under vacuum and the compound was purified by column chromatography affording the title compound (40 mg). 4H NMR (400 MHz, CD 3 OD): δ 8.85 (d, 1H, J = 2.0 Hz), 8.70 (d, 1H, J = 2.4 Hz), 8.22 (t, 1H, J = 2.0 Hz). ESI-MS: 172.0 [M + H] +. 28 PREPARATION OF (E) -5-CHLOR-N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) NICOTINOHYDRAZIDE.
5-chloronicotinohydrazide (30 mg, 0.175 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (29.8 mg, 0.175 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation to 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound as a solid (20 mg). NMR (400 MHz, acetone-d6): δ 9.06 (s, 1H), 8.77 (s, 1H), 8.37 (s, 1H), 7.62 (d, 1H, J = 2, 8 Hz), 7.31 (dd, 1H, J = 2.0 & 8.4 Hz), 6.95 (d, 1H, J = 8.8 Hz), 2.58 (s, 3H). ESI-MS: 324.0 [M + H] +. 29 PREPARATION OF (E) —Ν '- (1- (5-CHLOR-2-HYDROXYPHENYL) PROPYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
3- (morpholinosulfonyl) benzohydrazide (40 mg, 0.140 mmol) and 1- (5-chloro-2-hydroxyphenyl) propan-1-one (25.9 mg, 0.140 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound as a solid (20 mg). 1H-NMR (400 MHz, acetone): δ 8.26 (m, 2H), 8.00 (d, 1H, J = 7.6 Hz), 7.84 (t, 1H, J = 8.0 Hz), 7.64 (d, 1H, J = 2.4 Hz), 7.33 (m, 1H), 6.98 (d, 1H, J = 9.2 Hz), 3.69 (m, 4H), 3.10 (q, 2H, J = 7.6 Hz), 2.99 (m, 4H), 1.26 (t, 3H, J = 7.6 Hz). ESI-MS: 452.1 [M + H] +. 30 PREPARATION OF (E) -3- (MORPHOLINOSULPHONYL) -Ν '- (1- (PYRIDIN — 3 — IL) ETHYLIDENE) BENZOHIDRAZID.
3- (morpholinosulfonyl) benzohydrazide (40 mg, 0.140 mmol) and 1- (pyridin-3-yl) ethanone (16.98 mg, 0.140 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound as a solid (15 mg). 1H-NMR (400 MHz, CDCl3): δ 9.53 (bs, 1H), 8.87 (s, 1H), 8.59 (m, 1H), 8.39 (m, 1H), 8.17 ( m, 1H), 7.98 (m, 1H), 7.89 (d, 1H, J = 8.0 Hz), 7.67 (t, 1H, J = 8.0 Hz), 7.32 ( m, 1H), 3.70 (m, 4H), 3.00 (m, 4H), 2.39 (s, 3H). ESI-MS: 389.0 [M + H] +. 31. PREPARATION OF 3-MORPHOLINOBENZOHIDRAZIDE.
Methyl 3-morpholinobenzoate (100 mg, 0.452 mmol) was added to hydrazine (14.48 mg, 0.452 mmol) in methanol (10 mL) and refluxed for 12 hours at 65 ° C. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the compound was purified by column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound as a solid (52 mg). NMR (400 MHz, DMSO-d 6): δ 9.69 (s, 1H), 7.35 (s, 1H), 7.27 (m, 2H), 7.07 (m, 1H), 4.45 (bs, 2H), 3.74 (m, 4H), 3.14 (m, 4H). ESI-MS: 222.1 [M + H] +. 32 PREPARATION OF (E) -N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3-MORPHOLINOBENZOHIDRAZIDE.
1- (5-chloro-2-hydroxyphenyl) ethanone (40 mg, 0.234 mmol) and 3-morpholinobenzohydrazide (51.9 mg, 0.234 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the rational mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound (60 mg) as a solid. 1H-NMR (400 MHz, DMSO-d6): δ 7.65 (d, 1H, J = 2.4 Hz), 7.42-7.32 (m, 4H), 7.20 (m, 1H), 6.94 (d, 1H, J = 8.8 Hz), 3.77 (m, 4H), 3.19 (m, 4H), 2.48 (s, 3H). ESI-MS: 374.1 [M + H] +. 33 PREPARATION OF 5- (METHYLSULPHONYL) NICOTINOHYDRAZED.
Methyl 5- (methylsulfonyl) nicotinate (100 mg, 0.465 mmol) was added to hydrazine (17.87 mg, 0.558 mmol) in methanol (10 mL) and refluxed for 12 hours at 70 °. Ç. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the compound was purified by flash column chromatography (3% CH 3 OH / CH 2 Cl 2) to yield the title compound (83 mg, 80% yield). as a solid. 1 H NMR (400 MHz, CDCl 3): δ 9.20 (d, 1H, J = 2.0 Hz), 9.17 (d, 1H, J = 2.0 Hz), 8.61 (s, 1H) 3.11 (s, 3H). ESI-MS: 216.1 [M + H] +. 34 PREPARATION OF (E) -Ν '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -5- (METHYLSULPHONYL) NICOTINE HYDRAZIDE.
1- (5-chloro-2-hydroxyphenyl) ethanone (50 mg, 0.293 mmol) and 5- (methylsulfonyl) nicotine hydrazide (63.1 mg, 0.293 mmol) were dissolved in methanol (4 mL) in the presence of acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (3% CH 3 OH / CH 2 Cl 2) yielding the title compound (70 mg, 63.0% yield) as a solid. 1H-NMR (400 MHz, DMSO-d6): δ 11.86 (s, 1H), 9.37 (s, 1H), 9.27 (s, 1H), 8.76 (s, 1H), 7, 68 (s, 1H), 7.36 (d, 1H, J = 8.8 Hz), 6.97 (d, 1H, J = 8.8 Hz), 3.42 (s, 3H), 2, 53 (s, 3H). ESI-MS: 368.8 [M + H] +. 35 Preparation of 3- (Methylsulphonyl) BENZOHIDRAZED.
Methyl 3- (methylsulfonyl) benzoate (100 mg, 0.467 mmol) was added to hydrazine (22.44 mg, 0.700 mmol)) in methanol (10 mL) and refluxed for 12 hours at 70 ° C. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the compound was purified by flash column chromatography (3% CH 3 OH / CH 2 Cl 2) yielding the title compound (80 mg, 80% yield) as a solvent. solid. 1H-NMR (400 MHz, CDCl3): δ 8.28 (s, 1H), 8.07 (d, 1H, J = 7.6 Hz), 8.01 (d, 1H, J = Ί, ζ> Hz ), 7.62 (t, 1H, J = 7.6 Hz), 3.04 (s, 3H). ESI-MS: 215.1 [M + H] +. 36 PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3- (METHODSULPHONYL) BENZOHIDRAZIDE.
1- (5-chloro-2-hydroxyphenyl) ethanone (55 mg, 0.322 mmol) and 3- (methylsulfonyl) benzohydrazide (69.1 mg, 0.322 mmol) were dissolved in methanol (5 mL) in the presence of acid acetic acid as a catalyst, and the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (3% CH 3 OH / CH 2 Cl 2) yielding the title compound (75 mg, 63.4% of yield) as a solid. 1 H NMR (400 MHz, CD 3 OD): δ 8.49 (s, 1H), 8.26 (d, 1H, J = 8.4 Hz), 8.18 (d, 1H, J = 7.6 Hz) , 7.80 (t, 1H, J = 7.6 Hz), 7.60 (d, 1H, J = 2.4 Hz), 7.27 (m, 1H), 6.93 (d, 1H, J = 8.8 Hz), 3.19 (s, 3H), 2.49 (s, 3H). ESI-MS: 367.8 [M + H] +. 37. Preparation of 3 - ((4-Methylpiperidine-1-yl) sulfonyl) benzoic acid.
4-Methylpiperidine (180 mg, 1.813 mmol) was added to 3- (chlorosulfonyl) benzoic acid (200 mg, 0.906 mmol) in the presence of potassium carbonate (251 mg, 1.813 mmol) in THF (Volume: 5 mL ) at room temperature, and the reaction mixture was stirred for 12 h at room temperature. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (3% CH 3 OH / CH 2 Cl 2) yielded the title compound as a solid. 1 H NMR (CD 3 OD, 400 MHz): δ 8.32 (m, 1H), 8.27 (m, 1H), 7.96 (m, 1H), 7.72 (t, 1H, J = 8, Hz), 3.72 (m, 2H), 2.27 (m, 2H), 1.68 (m, 2H), 1.29 (m, 1H), 1.21 (m, 2H), 0 , 88 (d, 3H, J = 6.4 Hz). ESI-MS: 284.1 [M + H] +. 38. Preparation of Methyl 3 - ((4-methylpiperidine-1-yl) sulfonyl) benzoate.
3 - ((4-Methylpiperidin-1-yl) sulfonyl) benzoic acid (120 mg, 0.424 mmol) was refluxed in the presence of concentrated sulfuric acid (2.74 mg, 0.021 mmol) in methanol at 70 °. C, overnight. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography to yield methyl 3 - ((4-methylpiperidin-1-yl) sulfonyl) benzoate (100 mg, 0.319 mmol). 75% yield). 1H-NMR (CDCl3, 400 MHz): δ 8.39 (m, 1H), 8.25 (m, 1H), 7.94 (m, 1H), 7.62 (t, 1H, J = 7.6 Hz), 3.95 (s, 3H), 3.77 (m, 2H), 2.25 (m, 2H), 1.67 (m, 2H), 1.29 (m, 3H), 0, 90 (d, 3H, J = 4.8 Hz). ESI-MS: 298.1 [M + H] + 39. PREPARATION OF 2- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
Hydrazine (22.46 mg, 0.701 mmol) was added to methyl 2- (morpholinosulfonyl) benzoate (100 mg, 0.350 mmol) in methanol and refluxed for 12 hours at 70 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography, yielding the title compound 2- (morpholinosulfonyl) benzohydrazide (40 mg, 0.129 mmol, 36.8% yield) as a solid. 1H-NMR (CDCl3, 400 MHz): δ 7.86 (m, 1H), 7.66-7.56 (m, 2H), 7.52 (dd, 1H, J = 1.2 & 7.6 Hz ), 7.40 (m, 1H), 4.09 (m, 2H), 3.70 (m, 4H), 3.15 (m, 4H). ESI-MS: 286.1 [M + H] + 40. PREPARATION OF 3 - ((4-Methylpiperidine-1-yl) sulfonyl) benzodiazide.
Methyl 3 - ((4-methylpiperidin-1-yl) sulfonyl) benzoate (100 mg, 0.336 mmol) was added to hydrazine (21.55 mg, 0.693 mmol) in methanol and refluxed for 8 h 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography to yield 3 - ((4-methylpiperidin-1-yl) sulfonyl) benzohydrazide (70 mg, 0.217 mmol) with 64.4% yield). 1H-NMR (CD3OD, 400 MHz): δ 8.16 (m, 1H), 8.05 (m, 1H), 7.91 (m, 1H), 7.70 (t, 1H, J = 7.6Hz) ), 3.74 (m, 2H), 2.28 (m, 2H), 1.69 (m, 2H), 1.32-1.16 (m, 3H), 0.90 (d, 3H, J = 6.0 Hz). ESI-MS: 298.1 [M + H] + 41. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3 - ((4-Methylpiperidine-1-IL ) SULPHONY) BENZHYDRAZED.
[0281] 3 - ((4-methylpiperidin-1-yl) sulfonyl) benzohydrazide (70 mg, 0.235 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (40.2 mg, 0.235 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -N '- (1 - (5-chloro-2-hydroxyphenyl) ethylidene) -3 - ((4-methylpiperidin-1-yl) sulfonyl) benzohydrazide (15 mg, 0.032 mmol, 13.60% yield) as a solid. NMR (CDCl3, 400 MHz): δ 8.11 (m, 2H), 7.81 (m, 1H), 7.59 (m, 1H), 7.39 (m, 1H), 7.19 (m 1H), 6.89 (m, 1H), 3.69 (m, 2H), 2.41 (m, 2H), 2.24 (m, 2H), 1.63 (m, 2H), 1 .24 (m, 4H), 0.87 (d, 3H, J = 4.4 Hz). Mass [M + H] +: 450.2 42. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-FLUORPHENYL) ETHYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHIDRAZIDE.
1- (5-chloro-2-fluorophenyl) ethanone (20 mg, 0.116 mmol) and 3- (morpholinosulfonyl) benzohydrazide (33.1 mg, 0.116 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -N '- (1 - (5-chloro-2-fluorophenyl) ethylidene) -3- (morpholinosulfonyl) benzohydrazide (10 mg, 0.022 mmol, 19.22% yield) as a solid. 1 H NMR (CDCl 3, 400 MHz): δ 8.26 (m, 1H), 8.09 (m, 1H), 7.80 (d, 1H, J = 7.6 Hz), 7.58 (t, 1H, J = 7.6 Hz), 7.37 (m, 1H), 7.21 (m, 1H), 6.95 (m, 1H), 3.61 (m, 4H), 2.90 ( m, 4H), 2.29 (s, 3H). Mass [M + H] +: 440.1 43. PREPARATION DWE 3- (PYROLIDIN — 1 — ILSULFONYL) METHYL BENZOATE.
3- (Pyrrolidin-1-ylsulfonyl) benzoic acid (200 mg, 0.783 mmol) was refluxed in the presence of concentrated sulfuric acid (5.06 mg, 0.039 mmol) in methanol at 70 ° C overnight. the other. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography to afford methyl 3- (pyrrolidin-1-ylsulfonyl) benzoate (150 mg, 0.535 mmol, 68.3%). income). 1 H NMR (CDCl 3, 400 MHz): δ 8.47 (m, 1H), 8.25 (d, 1H, J = 7.6 Hz), 8.02 (dt, 1H, J = 1.2 &lt; 8.0 Hz), 7.63 (t, 1H, J = 7.6 Hz), 3.96 (s, 3H), 3.27 (m, 4H), 1.77 (m, 4H). Mass [M + H] +: 270.1 44. PREPARATION OF METHYL 3- (N-METHYLSULFAMOIL) BENZOATE.
3- (N-Methylsulfamoyl) benzoic acid (200 mg, 0,929 mmol) was refluxed in the presence of concentrated sulfuric acid (6.01 mg, 0.046 mmol) in methanol at 70 ° C overnight. the other. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography to yield methyl 3- (N-methylsulfamoyl) benzoate (120 mg, 0.497 mmol, 53.5% yield). ). 1 H NMR (CDCl 3, 400 MHz): δ 8.51 (m, 1H), 8.25 (m, 1H), 8.06 (dt, 1H, J = 1.2 & 8.0 Hz), 7, 63 (t, 1H, J = 7.6 Hz), 3.96 (s, 3H), 2.69 (s, 3H). Mass [M + H] +: 230.1 45. PREPARATION OF 3- (PYROLIDIN-1-ILSULFONYL) BENZOHIDRAZIDE.
Methyl 3- (pyrrolidin-1-ylsulfonyl) benzoate (150 mg, 0.557 mmol) was added to hydrazine (35.7 mg, 1.114 mmol) in methanol and refluxed for 12 hours at 65 ° C. After cooling, the reaction was monitored by TLC. After termination of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography to afford 3- (pyrrolidin-1-ylsulfonyl) benzohydrazide (110 mg, 0.396 mmol, 71.1% yield). ). 1 H NMR (CDCl 3, 400 MHz): δ 8.18 (m, 1H), 8.03 (d, 1H, J = 7.6 Hz), 7.97 (d, 1H, J = 8.0 Hz) ), 7.78 (bs, 1H), 7.63 (t, 1H, J = 7.6 Hz), 4.17 (bs, 2H), 3.25 (m, 4H), 1.77 (m , 4H). Mass [M + H] +: 270.1 46. PREPARATION OF 3- (HYDRAZINOCARBONYL) -N-METHYLBenzenesulfonamide.
Hydrazine (43.3 mg, 1.352 mmol) was added to methyl 3- (N-methylsulfamoyl) benzoate (155 mg, 0.676 mmol) in methanol and refluxed for 12 hours at 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography to afford 3- (hydrazinocarbonyl) -N-methylbenzenesulfonamide (120 mg, 0.502 mmol, 74.3% yield) . 1 H NMR (CDCl 3, 400 MHz): δ 8.25 (m, 1H), 8.01 (m, 2H), 7.64 (m, 2H), 4.63 (m, 1H), 4.17 ( m, 2H), 2.69 (d, 3H, J = 5.2 Hz). ESI-MS: 230.0 [M + H] + 47. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -2- (MORPHOLINOSULPHONYL) BENZOHYDRAZED
2- (morpholinosulfonyl) benzohydrazide (30 mg, 0.105 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (17.94 mg, 0.105 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -N '- (1 - (5-chloro-2-hydroxyphenyl) ethylidene) -2- (morpholinosulfonyl) benzohydrazide (10 mg, 0.022 mmol, 21.28% yield) as a solid. 1 H NMR (CD 3 OD, 400 MHz): δ 7.95 (d, 1H, J = 8.0 Hz), 7.95-7.70 (m, 2H), 7.66 (d, 1H, J = 7 , 6 Hz), 7.56 (d, 1H, J = 2.8 Hz), 7.25 (dd, 1H, J = 2.8 & 8.8 Hz), 6.91 (d, 1H, J = 8.4 Hz), 3.66 (m, 4H), 3.2 (m, 4H), 2.36 (s, 3H).
Mass [M + H] +: 438.1 48. PREPARATION OF (E) -3- (2- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) HYDRAZINOCARBONYL) —N — Methylbenzenesulfonamide.
[0288] 3- (hydrazinocarbonyl) -N-methylbenzenesulfonamide (120 mg, 0.523 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (89 mg, 0.523 mmol) were dissolved in methanol (volume: 4 mL) in presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -3- (2- (1- (5-chloro-2-hydroxyphenyl) ethylidene) hydrazinocarbonyl) -N-methylbenzenesulfonamide (75 mg, 0.192 mmol, in 36.8% yield) as a solid. 1 H NMR (CDCl 3, 400 MHz): δ 8.21 (m, 1H), 8.06 (m, 1H), 7.95 (d, 1H, J = 7.6 Hz), 7.59 (t, 1H, J = 8.0 Hz), 7.39 (d, 1H, J = 2.4 Hz), 7.18 (m, 1H), 6.90 (d, 1H, J = 8.0 Hz) 2.56 (s, 3H), 2.36 (s, 3H). Mass [M + H] +: 382.1 49. PREPARATION OF (E) —N '- (1- (5 — CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3- (PYROLIDIN-1-ILSULFONYL) BENZHYDRAZID.
[0289] 3- (pyrrolidin-1-ylsulfonyl) benzohydrazide (105 mg, 0.390 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (66.5 mg, 0.390 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -Ν '- (1 - (5-chloro-2-hydroxyphenyl) ethylidene) -3- (pyrrolidin-1-ylsulfonyl) benzohydrazide (70 mg, 0.163 mmol, 41.7% yield) as a solid. 1H-NMR (CDCl3, 400 MHz): δ 8.18 (m, 1H), 8.13 (m, 1H), 7.95 (d, 1H, J = 7.6 Hz), 7.65 (t, 1H, J = 7.6 Hz), 7.41 (m, 1H), 7.21 (m, 1H), 6.93 (d, 1H, J = 8.8 Hz), 3.23 (m, 4H), 2.39 (s, 3H), 1.75 (m, 4H).
Mass [M + H] +: 422.1 50. PREPARATION OF METHYL 3- (1,1-DIOXIDOTIOMORPHOLINE) BENZOATE.
3- (1,1-Dioxothiomorpholino) benzoic acid (100 mg, 0.392 mmol) was refluxed in the presence of concentrated sulfuric acid (2.53 mg, 0.020 mmol) in methanol (5 mL) at 70 ° C overnight. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography to afford methyl 3- (1,1-dioxothiomorpholino) benzoate (99 mg, 0.353 mmol, 90% yield). ). 1H-NMR (CDCl3, 400 MHz): δ 7.58 (m, 2H), 7.36 (t, 1H, J = 8.0 Hz), 7.09 (m, 1H), 3.91 (s, 3H), 3.89 (m, 4H), 3.11 (m, 4H). Mass [M + H] +: 270.1 51. PREPARATION OF 3- (1,1-DIOXIDOTIOMORPHOLINE) BENZOHIDRAZIDE.
Methyl 3- (1,1-dioxothiomorpholino) benzoate (95 mg, 0.353 mmol) was added to hydrazine (22.61 mg, 0.705 mmol) in methanol and refluxed for 12 hours at 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the title compound 3- (1,1-dioxothiomorpholino) benzohydrazide (32 ° C). mg, 0.109 mmol, 31.0% yield) as a solid. 1 H NMR (CDCl 3, 400 MHz): δ 7.34 (m, 1H), 7.29 (t, 1H, J = 8.4 Hz), 7.18 (d, 1H, J = 7.6 Hz ), 6.70 (dd, 1H, J = 4.8 & 8.0 Hz), 3.85 (m, 4H), 3.05 (m, 4H). Mass [M + H] +: 270.1 52. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3- (1,1-DIOXIDOTIOMORPHOLINE) BENZOHYDRAZED.
3- (1,1-dioxidothiomorpholino) benzohydrazide (30 mg, 0.111 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (19.00 mg, 0.111 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -N '- (1 - (5-chloro-2-hydroxyphenyl) ethylidene) -3- (1,1-dioxothiomorpholino) benzohydrazide (15 mg, 0.035 mmol, 31.3% yield) as a solid. 1H-NMR (DMSO-d6, 400 MHz): δ 7.65 (d, 1H, J = 2.0 Hz), 7.47 (m, 1H), 7.41 (t, 1H, J = 7.6 Hz), 7.36-7.27 (m, 3H), 6.94 (d, 1H, J = 8.8 Hz), 3.87 (m, 4H), 3.17 (m, 4H), 2.48 (s, 3H). Mass [M + H] +: 422.2 53. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-NITROPHENYL) ETHYLIDENE) -3- (MORPHOLINOSULPHONYL) BENZOHYDRAZIDE.
1- (5-chloro-2-nitrophenyl) ethanone (30 mg, 0.150 mmol) and 3- (morpholinosulfonyl) benzohydrazide (42.9 mg, 0.150 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the product (E) -N '- (1- ( 5-chloro-2-nitrophenyl) ethylidene) -3- (morpholinosulfonyl) benzohydrazide (15 mg, 0.030 mmol, 20.09% yield) as a solid. 1H-NMR (CDCl3, 400 MHz): δ 8.20 (m, 1H), 8.07 (m, 1H), 7.88 (m, 1H), 7.66 (m, 1H), 7.51 ( m, 2H), 7.39 (m, 1H), 3.69 (m, 4H), 2.99 (m, 4H), 2.29 (s, 3H). Mass [M + H] +: 468.0 54. PREPARATION OF METHYL 3-SULFAMOILBENZOATE.
3-Sulfamoylbenzoic acid (150 mg, 0.746 mmol) was refluxed in the presence of concentrated sulfuric acid (4.82 mg, 0.037 mmol) in methanol (5 mL) at 70 ° C overnight. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography which afforded methyl 3-sulfamoylbenzoate (115 mg, 0.524 mmol, 70.2% yield) as a solid. 1H-NMR (CDCl3, 400 MHz): δ 8.53 (m, 1H), 8.18 (d, 1H, J = 8.0 Hz), 8.08 (d, 1H, J = 7.6 Hz) 7.57 (t, 1H, J = 8.0 Hz), 3.92 (s, 3H). Mass [M + H] +: 216.0 55. PREPARATION OF METHYL 4- (MORPHOLINOSULPHONYL) BENZOATE.
4- (morpholinosulfonyl) benzoic acid (150 mg, 0.553 mmol) was refluxed in the presence of concentrated sulfuric acid (3.57 mg, 0.028 mmol) in methanol at 70 ° C overnight. The reaction was monitored by TLC.
After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography which afforded methyl 4- (morpholinosulfonyl) benzoate (135 mg, 0.464 mmol, 84% yield) . NMR (CDCl3, 400 MHz): δ 8.21 (m, 2H), 7.82 (m, 2H), 3.97 (s, 3H), 3.4 (m, 4H), 3.02 (m , 4H). Mass [M + H] +: 286.0 56. PREPARATION OF 3- (HYDRAZINOCARBONYL) BENZENOSULFONAMIDE.
Methyl 3-sulfamoylbenzoate (110 mg, 0.511 mmol) was added to hydrazine (32.8 mg, 1.022 mmol) in methanol and refluxed for 8 h at 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography (5% methanol / DCM) to afford 3- (hydrazinocarbonyl) benzenesulfonamide (57 mg, 0.260 mmol, 50%). 8% yield) as a white solid. 1 H NMR (CD 3 OD, 400 MHz): δ 8.32 (m, 1H), 8.04 (d, 1H, J = 7.6 Hz), 7.97 (d, 1H, J = 7.6 Hz) , 7.63 (t, 1H, J = 8.0 Hz). Mass [M + H] +: 216.07. Preparation of 4- (MORPHOLINOSULPHONYL) BENZOHYDRAZED.
Methyl 4- (morpholinosulfonyl) benzoate (135 mg, 0.473 mmol) was added to hydrazine (30.3 mg, 0.946 mmol) in methanol and refluxed for 8 h at 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography (3% methanol / DCM) to afford 4- (morpholinosulfonyl) benzohydrazide (102 mg, 0.350 mmol, 74, 0% yield) as a white solid. 1H-NMR (CDCl3, 400 MHz): δ 7.94 (m, 2H), 7.79 (m, 2H), 3.72 (m, 4H), 2.99 (m, 4H). Mass [M + H] +: 286.0 58. PREPARATION OF (E) -3- (2- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) HYDRAZINOCARBONYL) BENZENOSULFONAMIDE.
3- (hydrazinocarbonyl) benzenesulfonamide (50 mg, 0.232 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (39.6 mg, 0.232 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding product (E) -3- (2- (1 - (5-chloro-2-hydroxyphenyl) ethylidene) hydrazinocarbonyl) benzenesulfonamide (36 mg, 0.094 mmol, 40.4% yield) as a solid. NMR (DMSO-d 6, 400 MHz): δ 8.34 (s, 1H), 8.15 (d, 1H, J = 7.6 Hz), 8.02 (d, 1H, J = 7.6 Hz ), 7.73 (t, 1H, J = 8.0 Hz), 7.64 (m, 1H), 7.51 (bs, 2H), 7.32 (dd, 1H, J = 2.4 & 8.4 Hz), 6.92 (d, 1H, J = 8.4 Hz), 2.49 (s, 3H).
Mass [M + H] +: 368.0 59. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -4- (MORPHOLINOSULPHONYL) BENZOHYDRAZIDE.
4- (morpholinosulfonyl) benzohydrazide (100 mg, 0.350 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (59.8 mg, 0.350 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding (E) -N '- (1- (1 5-chloro-2-hydroxyphenyl) ethylidene) -4- (morpholinosulfonyl) benzohydrazide (80 mg, 0.177 mmol, 50.6% yield) as a solid. 7H NMR (DMSO-d6, 400 MHz): δ 8.16 (m, 2H), 7.89 (m, 2H), 7.67 (d, 1H, J = 2.4 Hz), 7.35 ( dd, 1H, J = 2.4 & 8.8 Hz), 6.95 (d, 1H, J = 8.4 Hz), 3.64 (m, 4H), 2.92 (m, 4H), 2.49 (s, 3H). Mass [M + H] +: 438.0 60. PREPARATION OF 3 - ((4-Methylpiperazin-1-yl) sulfonyl) benzoic acid.
3- (Chlorosulfonyl) benzoic acid (200 mg, 0.906 mmol) was added to 1-methylpiperazine (100 mg, 0.997 mmol) in the presence of potassium carbonate (251 mg, 1.813 mmol) in THF (volume: 5 mL ) at room temperature, and the reaction mixture was stirred for 12 h at room temperature. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (3% CH 3 OH / CH 2 Cl 2) yielded 3 - ((4-methylpiperazin-1-yl) sulfonyl acid product ) benzoic acid (100 mg, 0.320 mmol, 35.3% yield) as a solid. 1H-NMR (CDCl3, 400 MHz): δ 7.77 (m, 2H), 7.63-7.55 (m, 2H), 3.04 (m, 4H), 2.46 (m 4H), 2 .31 (s, 3H). Mass [M + H] +: 285.1 61. Preparation of Methyl 3 - ((4-methylpiperazin-1-yl) sulfonyl) benzoate.
3 - ((4-Methylpiperazin-1-yl) sulfonyl) benzoic acid (250 mg, 0.879 mmol) was refluxed in the presence of concentrated sulfuric acid (5.68 mg, 0.044 mmol) in methanol at 70 °. C overnight. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and the crude material was used for further reaction without purification. 62. Preparation of 3 - ((4-Methylpiperazin-1-yl) sulfonyl) benzohydrate.
Methyl 3 - ((4-methylpiperazin-1-yl) sulfonyl) benzoate (200 mg, 0.670 mmol) was added to hydrazine (43.0 mg, 1.341 mmol) in methanol and refluxed for 8 h 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography (3% methanol / DCM) to afford 3 - ((4-methylpiperazin-1-yl) sulfonyl) benzohydrazide (125 mg, 0.406 mmol, 60.6% yield) as a white solid. 1 H NMR (DMSO-d 6, 400 MHz): δ 10.08 (s, 1H), 8.12 (m, 2H), 7.84 (d, 1H, J = 7.6 Hz), 7.72 ( t, 1H, J = 7.6 Hz), 4.57 (m, 1H), 2.88 (m, 4H), 2.32 (m, 4H), 2.10 (s, 3H). Mass [M + H] +: 298.9 63. PREPARATION OF (E) —N '- (1- (5 — CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3 - ((4-METHYPYPERAZIN-1-IL) SULFONY ) BENZOHYDRAZED.
[0303] 3 - ((4-methylpiperazin-1-yl) sulfonyl) benzohydrazide (85 mg, 0.285 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (48.6 mg, 0.285 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. Upon completion of the reaction, after cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) afforded the product (E) -Ν '- (1- ( 5-chloro-2-hydroxyphenyl) ethylidene) -3 - ((4-methylpiperazin-1-yl) sulfonyl) benzohydrazide (70 mg, 0.152 mmol, 53.4% yield) as a solid. 1 H NMR (CD 3 OD, 400 MHz): δ 8.29 (s, 1H), 8.21 (d, 1H, J = 7.2 Hz), 7.99 (d, 1H, J = 8.0 Hz) , 7.78 (t, 1H, J = 7.6 Hz), 7.59 (d, 1H, J = 2.4 Hz), 7.27 (dd, 1H, J = 2.4 & 9.2 Hz), 6.92 (d, 1H, J = 8.8 Hz), 3.09 (m, 4H), 2.54 (m, 4H), 2.48 (s, 3H), 2.28 ( s, 3H). Mass [M + H] +: 450.9 64. PREPARATION OF 3- (PIPERIDIN-1-ILSULFONYL) BENZOHIDRAZIDE.
Methyl 3- (piperidin-1-ylsulfonyl) benzoate (150 mg, 0.529 mmol) was added to hydrazine (50.9 mg, 1.588 mmol) in methanol and refluxed for 8 h at 65 ° C. After cooling, the reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by flash chromatography (3% methanol / DCM) to afford 3- (piperidin-1-ylsulfonyl) benzohydrazide (70 mg, 0.245 mmol, 46.2% yield) as a white solid. 1H-NMR (CD3OD, 400 MHz): δ 8.17 (t, 1H, J = 1.2 Hz), 8.05 (dt, 1H, J = 1.2 & 8.0 Hz), 7.90 ( dt, 1H, J = 1.2 & 8.0 Hz), 7.69 (t, 1H, J = 7.6 Hz), 2.99 (m, 4H), 1.62 (m, 4H), 1.43 (m, 2H). Mass [M + H] +: 284.1 65. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -3- (PIPERIDIN-1-ILSULFONYL) BENZOHIDRAZIDE.
[0305] 3- (piperidin-1-ylsulfonyl) benzohydrazide (65 mg, 0.229 mmol) and 1- (5-chloro-2-hydroxyphenyl) ethanone (39.1 mg, 0.229 mmol) were dissolved in methanol (volume: 4 mL) in the presence of acetic acid as a catalyst and then the reaction mixture was heated by microwave irradiation at 120 ° C for 30 min. The reaction was monitored by TLC. Upon completion of the reaction, after cooling, the solvent was removed under vacuum and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) yielding the product (E) -N '- (1- (5-chloro-2-hydroxyphenyl) ethylidene) -3- (piperidin-1-ylsulfonyl) benzohydrazide (55 mg, 0.124 mmol, 53.9% yield) as a solid. 1 H NMR (CDCl 3, 400 MHz): δ 8.09 (m, 2H), 7.85 (d, 1H, J = 8.0 Hz), 7.62 (t, 1H, J = 8.0 Hz) , 7.41 (d, 1H, J = 2.4 Hz), 7.22 (d, 1H, J = 8.0 Hz), 6.93 (d, 1H, J = 8.8 Hz), 2 .97 (m, 4H), 2.41 (s, 3H), 1.61 (m, 4H), 1.40 (m, 2H). Mass [M + H] +: 436.9 66. Preparation of 4-Chloro-2- (4-Methyl-1H-PIRAZOL-5-IL) PHENOL.
A mixture of (E) -3- (5-chloro-2-hydroxyphenyl) -2-methylacrylaldehyde (40 mg, 0.203 mmol) and 4-methylbenzenesulfonohydrazide (41.7 mg, 0.224 mmol) in acetonitrile ( 3 mL) was stirred at room temperature for 3 h and then acetonitrile (2 mL) and sodium hydroxide (8.95 mg, 0.224 mmol) were added, and the mixture was heated at reflux for 16 h. The product was used for further reaction without purification. 67. PREPARATION OF 3- (MORPHOLINOSULPHONYL) BENZOIC ACID.
3- (Chlorosulfonyl) benzoic acid (250 mg, 1.133 mmol) was added to morpholine (99 mg, 1.133 mmol) in the presence of potassium carbonate (313 mg, 2.266 mmol) in THF (5 mL) at room temperature. , and the reaction mixture was stirred for 12 h at room temperature. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (3% CH 3 OH / CH 2 Cl 2) yielding the title compound (160 mg) as a solid. 1 H NMR (400 MHz, CD 3 OD): δ 8.34 (m, 1H), 8.32 (d, 1H, J = 8.0 Hz), 7.99 (m, 1H), 7.76 (t, 1H, J = 8.0 Hz), 3.70 (m, 4H), 2.98 (m, 4H). ESI-MS: 272.0 [M + H] + 68. PREPARATION OF (3- (5-CHLOR-2-HYDROXYPHENYL) —4-METHYL-1H-PIRAZOL-1-IL) (3- (MORPHOLINOSULPHONYL) PHENYL) Methanone.
A mixture of (E) -3- (5-chloro-2-hydroxyphenyl) -2-methylacrylaldehyde (40 mg, 0.203 mmol) and 4-methylbenzenesulfonohydrazide (41.7 mg, 0.224 mmol) in acetonitrile ( 3 mL) was stirred at room temperature for 3 h and then acetonitrile (2 mL) and sodium hydroxide (8.95 mg, 0.224 mmol) were added, and the mixture was heated at reflux for 16 h then sodium hydroxide (12.21 mg, 0.305 mmol) and 3- (morpholinosulfonyl) benzoyl chloride (88 mg, 0.305 mmol) (made of 3- (morpholinosufonyl) benzoic acid) were then added, and the mixture was stirred at room temperature. room for 2 h. The reaction was monitored by TLC. After completion of the reaction, the product was extracted with EtOAc and the organic layer was washed with brine, dried over anhydrous Na 2 SO 4 and filtered, and the solvent removed under vacuum. The resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2) and yielded the desired product (3- (5-chloro-2-hydroxyphenyl) -4-methyl-1H-pyrazol-1-yl) ( 3- (morpholinosulfonyl) phenyl) methanone (30 mg, 0.064 mmol, 31.3% yield) as a solid. 1 H NMR (400 MHz, CDCl 3): δ 8.28 (m, 2H), 8.20 (d, 1H, J = 8.0 Hz), 7.95 (d, 1H, J = 8.4 Hz) , 7.70 (t, 1H, J = 7.6 Hz), 7.59 (d, 1H, J = 2.4 Hz), 7.18 (dd, 1H, J = 2.8 & 8.8 Hz), 6.87 (d, 1H, J = 8.4 Hz), 3.68 (m, 4H), 3.02 (m, 4H), 2.40 (s, 3H). ESI-MS: 462.0 [M + H] + 69. PREPARATION OF 4-CHLOR-2- (1H-PIRAZOL-3-IL) PHENOL
A mixture of (E) -3- (5-chloro-2-hydroxyphenyl) -2-methylacrylaldehyde (40 mg, 0.203 mmol) and 4-methylbenzenesulfonhydrazide (41.7 mg, 0.224 mmol) in acetonitrile ( 3 mL) was stirred at room temperature for 3 h and then acetonitrile (2 mL) and sodium hydroxide (8.95 mg, 0.224 mmol) were added, and the mixture was heated at reflux for 16 h. The product was used for further reaction without purification. 70. PREPARATION OF (3- (5-CHLOR-2-HYDROXYPHENYL) -1H-PIRAZOL-1-IL) (3- (MORPHOLINOSULPHONYL) PHENYL) METHONE.
3- (morpholinosulfonyl) benzoic acid (50mg, 0.184 mmol), β-benzo [d] [1,2,3] triazol-1-ol (37.4 mg, 0.276 mmol), EDC (53.0 mg, 0.276 mmol) and sodium bicarbonate (17.03 mg, 0.203 mmol) were dissolved in THF (10 mL) and then 4-chloro-2- (1H-pyrazol-3-yl) phenol (35, 9 mg, 0.184 mmol) was added at room temperature, and the reaction was stirred overnight at room temperature. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (3- (5-chloro-2-hydroxyphenyl) -1H-pyrazol-1-yl) (3- (morpholinosulfonyl) phenyl) methanone (43 mg, 0.094 mmol, 51.0% yield) as a solid. 3 H NMR (400 MHz, CDCl 3): δ 8.68 (s, 1H), 8.39 (d, 1H, J = 7.6 Hz), 8.01 (d, 1H, J = 7.6 Hz) , 7.80 (d, 1H, J = 2.4 Hz), 7.70 (t, 1H, J = 7.6 Hz), 7.53 (d, 1H, J = 2.4 Hz), 7 , 38 (dd, 1H, J = 2.4 & 8.4 Hz), 7.27 (d, 1H, J = 8.4 Hz), 6.52 (s, 1H), 3.75 (m, 4H), 3.05 (m, 4H). ESI-MS: 448.0 [M + H] + 71. PREPARATION OF 5 - ((4-Methylpiperazin-1-yl) sulfonyl) NICOTINOIDRAZIDE.
Hydrazine (11.78 mg, 0.367 mmol) was added to methyl 5 - ((4-methylpiperazin-1-yl) sulfonyl) nicotinate (55 mg, 0.184 mmol) in methanol (10 mL) and was subjected to reflux overnight. The reaction was monitored by TLC, after completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (3% methanol / DCM) yielding the title compound 5 - ((4 -methylpiperazin-1-yl) sulfonyl) nicotinohydrazide (45 mg, 0.147 mmol, 80% yield) as a solid. 1 H NMR (400 MHz, CDCl 3): δ 9.07 (s, 1H), 9.02 (s, 1H), 8.33 (s, 1H), 7.46 (bs, 1H), 4.09 ( bs, 2H), 3.05 (m, 4H), 2.43 (m, 4H), 2.21 (s, 3H). ESI-MS: 300.1 [M + H] + 72. PREPARATION OF (E) —N '- (1- (5 — CHLOR-2 — HYDROXYPHENYL) ETHYLIDENE) -5 - ((4-METHYPIPERAZIN-1-IL ) SULFONY) NICOTINOHYDRAZED
1- (5-chloro-2-hydroxyphenyl) ethanone (25.07 mg, 0.147 mmol) and 5 - ((4-methylpiperazin-1-yl) sulfonyl) nicotinehydrazide (40 mg, 0.134 mmol) were dissolved in methanol (10 mL) in the presence of acetic acid as catalyst and then the reaction mixture was refluxed for 12 hours at 70 ° C. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -N '- (1 - (5-chloro-2-hydroxyphenyl) ethylidene) -5 - ((4-methylpiperazin-1-yl) sulfonyl) nicotinehydrazide as a solid. 1 H NMR (400 MHz, CDCl 3): δ 9.21 (s, 1H), 9.00 (s, 1H), 8.48 (s, 1H), 7.41 (d, 1H, J = 2.0 Hz), 7.20 (d, 1H, J = 8.4 Hz), 6.90 (d, 1H, J = 8.4 Hz), 3.08 (m, 4H), 2.48 (m, 4H), 2.40 (s, 3H), 2.25 (s, 3H). ESI-MS: 452.0 [M + H] + 73. PREPARATION OF 5- (MORPHOLINOSULPHONYL) NICOTINATE
METALA
Methyl 5- (chlorosulfonyl) nicotinate (35 mg, 0.149 mmol) was added to morpholine (25.9 mg, 0.297 mmol) in the presence of potassium carbonate (41.1 mg, 0.297 mmol) in THF (8 mL) at room temperature and the reaction mixture was stirred for 12 hours at room temperature. The reaction was monitored by TLC, after completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (2% methanol / DCM) yielding 5- (morpholinosulfonyl) nicotinate product. of methyl (26 mg, 0.090 mmol, 60.5% yield) as a solid. 1H-NMR (400 MHz, CDCl3): δ 9.41 (s, 1H), 9.12 (s, 1H), 8.60 (d, 1H, J = 2.0 Hz), 4.01 (s, 3H), 3.76 (m, 4H), 3.07 (m, 4H). 74. PREPARATION OF 5- (MORPHOLINOSULPHONYL) NICOTINOHYDRAZED
Hydrazine A (5.60 mg, 0.175 mmol) was added to methyl 5- (morpholinosulfonyl) nicotinate (25 mg, 0.087 mmol) in methanol (10 mL) and refluxed overnight. The reaction was monitored by TLC, after completion of the reaction, the solvent was removed under vacuum and then the compound was purified by column chromatography (3% methanol / DCM) yielding the title compound 5- (morpholinosulfonyl). nicotinohydrazide as a solid. 1H-NMR (400 MHz, CDCl3): δ 9.15 (s, 1H), 8.98 (s, δ1), 8.41 (s, 1H), 3.71 (m, 4H) 3.02 ( m, 4H) 75. PREPARATION OF (E) —N '- (1- (5-CHLOR-2-HYDROXYPHENYL) ETHYLIDENE) -5- (MORPHOLINOSULPHONYL) NICOTINOHYDRAZIDE.
1- (5-chloro-2-hydroxyphenyl) ethanone (6.55 mg, 0.038 mmol) and 5- (morpholinosulfonyl) nicotine hydrazide (10 mg, 0.035 mmol) were dissolved in methanol (3 mL), in presence of acetic acid as catalyst and then the reaction mixture was refluxed for 12 hours at 70 ° C. The reaction was monitored by TLC. After completion of the reaction and cooling, the solvent was removed under vacuum, and the resulting crude material was purified by flash column chromatography (2% CH 3 OH / CH 2 Cl 2), yielding the title compound (E) -N '- ( 1- (5-chloro-2-hydroxyphenyl) ethylidene) -5- (morpholinosulfonyl) nicotine hydrazide (10 mg, 0.023 mmol, 65.2% yield) as a solid. 1 H NMR (400 MHz, DMSO-d 6): δ 11.82 (bs, 1H), 9.35 (s, 1H), 9.08 (s, 1H), 8.52 (s, 1H), 7, 66 (s, 1H), 7.34 (d, 1H, J = 8.4 Hz), 6.94 (d, 1H, J = 8.8 Hz), 3.63 (m, 4H), 2, 99 (m, 4H), 2.50 (s, 3H). ESI-MS: 439.1 [M + H] +.
76. GENERAL BIOCHEMISTRY AND CELLULAR METHODS AND MATERIALS
LSD1 activity was determined using a scan assay LSD1 inhibitor kit (Cayman Chemical Item No. 700120) purchased from Cayman Chemical Company (Ann Arbor, Michigan). Recombinant monoamine oxidase A and monoamine oxidase B (expressed in baculovirus infected BTI insect cells) (Catalog No. M7316 and M7441, respectively) were purchased from Sigma-Aldrich Co. LLC. (St. Louis, Missouri). The MAO-Glo ™ Assay Kit was purchased from Promega Corporation (Madison, Wisconsin). An ATPlite ™ luminescence assay system (e.g., Catalog No. V1401) was purchased from PerkinElmer Inc. (Waltham, Massachusetts).
77. CELL CULTURE
[0317] Cancer cell lines were obtained from the ATCC. Cells were cultured according to the procedures provided. Cell lines included those shown in Table 4 below. In addition to the supplements listed in Table 4, the media were also supplemented with 1% penicillin / streptomycin (100 IU / mL penicillin and 100 gg / mL streptomycin). Cells were cultured at 37 ° C and 5% CO 2. ATCC is the American Cell Culture Collection (Manassas, Virginia). * All organ / tissue sources were of human origin. ** FCS is fetal calf serum *** Derived from the MCF7 cell line characterized by non-nuclear estrogen receptor and high HER2 levels (Massarweh S, et al. (2008) Cancer Research 68: 826-33). 78. LSD1 HISTONE DEMETILASE ASSAY [0318] The primary assay for inhibitory activity of the compound was the LSD1 inhibitor scan assay kit (Cayman Chemical Company, Ann Arbor, Michigan; Cayman Chemical Item No. 700120). In sum, the test compounds were diluted to 20x the desired test concentration in 100% DMSO and 2.5 µl of drug drug sample were added to a 384-well black lacquer. The LSD1 enzyme stock was diluted 17-fold with assay buffer and 40 μΜ of diluted LSD1 enzyme was added to the appropriate wells. The reaction mixture comprised horseradish peroxidase, dimethyl peptide K4 (corresponding to the first 21 amino acids of the N-terminal histone H3 tail) and 10-acetyl-3,7-dihydroxyphenoxazine was then added to the wells. Resorufin generation (generated by reaction with H2O2 produced in the reaction) was analyzed on an Envision microplate reader with 530 nm excitation wavelength and 595 nm emission wavelength. 79. MONOAMINE OXIDASE ("MAO") TEST [0319] Inhibition of monoamine oxidase activity was performed using the MAO-Glo ™ assay kit according to the manufacturer's suggested protocol. In short, 6.25 µl of test compound was added to each well of a 384-well plate. Enzyme (MAO A or B) was added (12.5 pL in 2x buffer containing 1 pg protein) and allowed to incubate for 5 minutes. Finally, 6.25 pL of 4x MAO substrate was added to each well. After a one hour incubation, 25 µl luciferin detection reagent was added to each well and incubated for 20 minutes. Luminescence was then measured on an Envision microplate reader. Representative data used to determine the IC 50 for inhibition of each MAO isoform is provided in Figure 4, and representative data for various compounds are summarized in Table 8 below.
80. CELL VIABILITY TEST
Cell viability was determined using the ATPlite ™ luminescence assay system (PerkinElmer Inc., Waltham, Massachusetts) using the various cell lines described above and table 4. In short, cells were seeded in 96-well plates and then treated with different inhibitor concentrations (0.1% final DMSO concentration). After 96 hours of incubation, ATPlite detection reagent was added directly to the cell culture. Luminescence was read 5 minutes later on an Envision microplate reader. Representative IC50 data for inhibition of cell growth with various cell lines are provided below in Tables 6, 7 and 9.
81. Real-Time PCR
In short, T-47D cells were seeded in 96-well plates and treated with inhibitor concentrations as indicated. Single-color cell lysates, reverse transcription and real-time PCR were performed using the Cells-to-Ct kit (Life Technologies). Heme oxygenase (HMOX) transcript levels were normalized to hypoxanthine phosphoribosyltransferase (HPRT) and β-actin. Primers used in real time PCR are shown below in Table 5, and representative data for the effect of the disclosed compounds on HMOX expression are provided in Tables 6 and 7. TABLE 5.
Designation Amplification Target Initiator Sequence HMOX_F Heme Oxygenase AACTTTCAGAAGGGCCAGGT
HMOX_R Heme oxygenase GTAGACAGGGGCGAAGACTG
HPRT_F Hypoxanthine TGCTGAGGATTTGGAAAGGGTG phosphoribosyltransferase Designation Amplification Target Primer Sequence HPRT_R Hypoxanthine C C T T GAGCACACAGAGGGGC TAC phosphoribosyltransferase B-Actin_F β-actin CTGAGGACGACT
B-Actin_R β-actin AAGGGACTTCCTGTAACAACGCA 82. IC50 calculation [0322] IC50 values are determined using GraphPad Prism 5 software. Data were entered as an X-Y graph in the software as the percent inhibition for each drug concentration. Drug concentration values were transformed to the logarithmic scale and nonlinear regression was performed using the "sigmoidal dose response (variable slope)" option in GraphPad software to model the data and calculate IC50 values.
Reported IC50 values are the drug concentration at which 50% inhibition was achieved.
83. COMPOUND ACTIVITY
The ability of representative disclosed compounds to modulate various biochemical and cellular activities was determined using the assays described above. Results are shown in the tables below. The IC50 (μΜ) for inhibition of LSD1 activity or cell growth using T-47D cells is shown in Tables 6 and 7. In addition, the effect of representative compounds on heme oxygenase (HMOX) expression is also shown in Tables 6 and 7. The IC50 for inhibition of monoamine oxidases A ("MAO A") and B ("MAO B") by representative compounds compared to a compound control, tranylcypromin, is shown in Table 8. The effect of compound No 12. (referring to the compound number used in Table 7, or (E) -N '- (1- (5-chloro-2-hydroxyphenyl) ethylidene) -3- (morpholinosulfonyl) benzohydrazide) in cell growth for various strains If an IC50 or other assay result is indicated as "nd", it was not determined in the indicated assay.
Compound 12 was used to assess sensitivity in a panel of cancer cell lines (Table 9). Cell line sensitivity to compound 12 in this viability assay ranged by a logarithm with IC50 values around 300 nM to just under 3 μΜ. For comparison between representative compounds, IC 50 values were determined in T-47D cells (see tables 6 and 7). With few exceptions, T-47D cells were found to be sensitive to test compounds that were active in the LSD1 biochemical assay, and were less sensitive to compounds that demonstrated less activity in the LSD1 assay.
To add an extra level of analysis of LSD1 inhibition in cell culture by these compounds, expression matrix experiments were performed to evaluate the transcriptional changes induced by compound 12 (data not shown). These data indicated that heme oxygenase 1 (HMOX1) was one of the most consistently overregulated genes among multiple cell lines following treatment with this compound. As HMOX1 is known to be regulated by H3 methylation in the promoter (Krieg, AJ, et al. Mol Cell Biol 2010, 30 (1), 344-53), the effect of test compounds on HMOX1 expression in T cells -47D has been determined (see tables 6 and 7). The data show that representative compounds that are associated with overregulation of HMOX1 expression are also associated with inhibitory activity in the LSD1 assay and the cell viability assay.
LSD1 has a high structural homology to the monoamine oxidase family of enzymes (17.6% for both monoamine oxidase A and B; MAO-A and B, respectively; for example, see Gooden, DM, et al. Bioorg Med Chem Lett 2008, 18 (10), 3047-51). Selective activity of representative compounds for LSD1 compared to MAO A or MAO B is a desirable property for therapeutic compounds targeting LSD1. The specificities of compound 1 and compound 12 were tested in the MAO biochemical assays described herein (see Figure 3 for representative results which are summarized in table 8). In this assay, the known MAO inhibitor tranilcipromin showed activity against MAO A and B. In contrast, compound 1 exhibited activity comparable to tranilcipromin in relation to MAO B but showed no activity against MAO A. However , compound 12 showed no activity against any MAO enzyme (> 300μΜ). Compounds 18 and 24 were also tested and showed no activity against MAO A or B, and the results are given in Table 8. These results demonstrate that representative compounds have specificity for LSD1 with significantly reduced effect on MAO enzymes. It should be noted that both MAO A and MAO B differ from LSD1 in that FAD is covalently linked to the enzyme via a thioether bond with Cys406 and Cys397, respectively (Kearney, EB, et al. European Journal of Biochemistry 1971, (24), 321-327, and Bach, AW, et al Proc Natl Acad Sci USA 1988, (85), 4934-4938).
Table 6
No. Structure Activity Growth LSD1 expression, cellular, HMOX (times induction IC50 (pM) IC50 (pM)) - δ 0.218 2/7
Lineage Cellular Cell Growth, IC50 (μΜ) MDA-MB-235 0, 728 MDA-MB-435 1,440 MDA-MB-468 2,730 MIA PaCa-2 0,468 PANC-1 1,104 PC-3 2,160 SK-N-MC 0,329 T- 47D 0,649 U87 1,160 84. IN VIVO TUMOR EFFECTS FORECAST: MODEL
XENOXERT OF CELL LINEAGE
The following examples of the in vivo effect of the disclosed compounds are predictions. Generally, agents that modulate chromatin regulation, including histone demethylase inhibitors, exhibit efficacy in preclinical cancer models. The in vivo effects of the compounds described in the previous examples are expected to be demonstrated in various animal cancer models known to the person skilled in the art, such as tumor xenograft models. These models are typically performed on rodents, most often in mice, but can be performed on other animal species, as is convenient for the purposes of the study. The compounds, products and compositions disclosed herein are expected to demonstrate in vivo effects on various animal cancer models known to the person skilled in the art, such as mouse tumor xenograft models.
The in vivo effects of the compounds can be evaluated with a mouse tumor xenograft study, and a possible study protocol described therein. In short, cells (2 to 5 χ 106 in 100 mL of culture medium) were implanted subcutaneously, for example by subcutaneous injection, into the right rear flank of athymic nude / nude mice (5 to 6 weeks of 18 to 22 g). For the test compounds of the present invention, a typical cell line used for tumor xenograft study would be AN3 CA or BT-20. Other suitable cell lines for these studies are BT-549, HCT 116, HER218, MCF7, MDA-MB-231, MDA-MB-235, MDA-MB-435S, MDA-MB-468, PANC-1, PC cells. -3, SK-N-MC, T-47D and U-87 MG.
Cells are cultured prior to collection for this protocol as described in the present invention.
Following implantation, tumors are allowed to grow to about 100 mm3, usually about 6 to 18 days after implantation, before animals are randomly assigned to treatment groups (eg vehicle, positive control and multiple doses). of the test compound); the number of animals per group is typically 8 to 12. Day 1 of the study is the day animals receive their first dose. The effectiveness of a test compound can be determined in studies of various durations, according to the study objectives. Typical study periods are 14, 21 and 28 days. Dosing frequency (for example, whether animals are dosed with the test compound daily, every two days, every three days or at other frequencies) is determined for each study, depending on the toxicity and potency of the test compound. A typical study design would involve daily dosing (M-F) with the test compound with weekend recovery. Throughout the study, tumor volumes and body weights are measured twice a week. At the end of the study, the animals are euthanized and the tumors collected and frozen for further analysis. Alternatively, tumors may be processed immediately for analysis, for example, buffered formalin fixed, paraffin embedded and sectioned for hematoxylin / eosin staining, and further subjected to immunohistochemical analysis for the desired oncology markers.
For example, the compounds of the invention, or a pharmaceutically acceptable salt, solvate, polymorph, hydrate and stereochemically isomeric form thereof, are expected to demonstrate such effects in vivo.
85. IN VIVO ANTI-TUMOR EFFECTS FORECAST: MODEL
TUMOR
Alternatively, it may be desirable to evaluate the in vivo efficacy of the disclosed compounds in a tumor explant or in tumor graft animal models (for example, see Rubio-Viqueira B., et al. Clin Cancer Res. (2006) 12). : 4652-4661; Fiebig, HH, Maier, A. and Burger, AM EUR J. J. Canc. (2004) 40: 802-820; and DeRose, YS, et al. "Patient-derived tumor grafts authentically reflect tumor pathology , growth, metastasis and disease outcomes. "(2011) Nat. Med., printed). These models may provide higher quality information on the in vivo effects of therapeutic compounds. Tumor graft models are believed to be the most authentic in vivo models of many types of cancer, for example human breast cancer, with which to examine tumor biology and how they metastasize. Grafting effective patient tumor tissues in immunodeficient mice (called "tumor grafts") provides improvements over cell line implantation in terms of human tumor phenocopy and prediction of drug responses in patients (Clarke, R. Breast Cancer Res (2009) 11 Suppl 3, S22; Press, JZ, et al., Gynecol Oncol (2008) 110: 56-264; Kim, MP, et al., Nat. Protoc (2009) 4: 670-1680; Daniel, VC, et al., Cancer Res (2009) 69: 3364-3373; and Ding, L., et al., Nature (2010) 464: 999-1005).
[0332] In short, tissue samples will be collected from informed patients who have provided consent at Huntsman Cancer Hospital / University of Utah under an approved IRB protocol. Samples will be collected and identified by the Huntsman Cancer Institute's Tissue Resource and Application Core unit before they are obtained for deployment. All primary tumors are expected to come from individuals who did not receive chemotherapy prior to tissue collection, and all metastatic effusions will come from individuals who had been treated with chemotherapy, hormone therapy and / or radiotherapy. The University of Utah Institutional Animal Care and Use Committee will review and approve all mouse experiments. It is anticipated that a minimum of three mice per experimental group will be used, and only female mice will be used for studies involving breast cancer tumors. A single fresh or frozen tumor fragment (~ 8 mm3), or about 106 cells in matrigel, is implanted in the inguinal mammary fat of 3-4 week old female NOD / SCID mice. At the same time, interscapular estrogen pellets are implanted subcutaneously in mice with ER + tumors. Tumor growth is measured weekly using calipers. When tumors reach about 150 to 2,000 mm3, mice are sacrificed, and tissue fragments are retransplanted into another group of mice, frozen for later use and / or analyzed for histological, gene expression and DNA copy number. . Tumor volumes are calculated using the formula 0.5 χ length χ (width) 2. For the experiments to determine estrogen dependence, ER + tumors are implanted in mice, as described above, in the presence or absence of intraescapular estrogen pellets and with or without a simultaneous surgical procedure to remove the ovaries, which is performed according to the methods. standard.
[0333] Newly collected tumor tissues from patients or mice are cut into ~ 8 mm3 pieces and stored in liquid nitrogen in a 95% FBS and 5% DMSO solution for subsequent implantation. Alternatively, tissue is digested with collagenase solution (1 mg / mL collagenase [type IV, Sigma] in RPMI 1640 supplemented with 2.5% FBS, 10 mM HEPES, 10 μg / mL penicillin-streptomycin) at 37 ° C. ° C for 40 to 60 min while stirring at 250 rpm. Digested tissue is filtered to remove debris and washed in human breast epithelial cell (HBEC) medium (DMEM F / 12 supplemented with 10 mM HEPES, 5% FBS, 1 mg / mL BSA, 0.5 pg / mL hydrocortisone , 50 pg / ml gentamycin and 1 pg / ml ITS-X100) three times. The pellet is resuspended in freezing medium (5% FBS and 10% DMSO in HBEC medium) and stored in liquid nitrogen.
To assess the effect of a disclosed compound, tumors in mice are allowed to grow to about 100 mm3, usually about 6 to 18 days after implantation, before animals are randomly assigned to treatment groups (e.g., vehicle, positive control and various doses of test compound); the number of animals per group is typically 8 to 12. Day 1 of the study is the day animals receive their first dose. The effectiveness of a test compound can be determined in studies of various durations, according to the study objectives. Typical study periods are 14, 21 and 28 days. Dosing frequency (for example, whether animals are dosed with the test compound daily, every two days, every three days or at other frequencies) is determined for each study, depending on the toxicity and potency of the test compound. A typical study design would involve daily dosing (M-F) with the test compound with weekend recovery. Throughout the study, tumor volumes and body weights are measured twice a week. At the end of the study, the animals are euthanized and the tumors collected and frozen for further analysis. Alternatively, tumors may be processed immediately for analysis, for example, buffered formalin fixed, paraffin embedded and sectioned for hematoxylin / eosin staining, and further subjected to immunohistochemical analysis for the desired oncology markers.
For example, the compounds of the invention, or a pharmaceutically acceptable salt, solvate, polymorph, hydrate and stereochemically isomeric form thereof, are expected to demonstrate such effects in vivo.
86. EXAMPLES OF PHARMACEUTICAL COMPOSITIONS PROVIDED
"Active ingredient" as used in all these examples refers to one or more of the compounds of the invention, or a pharmaceutically acceptable salt, solvate, polymorph, hydrate, and stereochemically isomeric form thereof. The following examples of formulation of the compounds of the present invention in tablets, suspension, injectables and ointments are predictions.
Typical examples of recipes for the formulation of the invention are provided below. Various other dosage forms may be applied in the present invention, such as a filled gelatin capsule, liquid emulsion or suspension, ointments, suppositories or a chewable tablet form, employing the disclosed compounds in desired dosage amounts according to the present invention. Various conventional techniques for preparing suitable dosage forms may be used to prepare the intended pharmaceutical compositions, such as those disclosed herein and in standard reference texts, for example, in the British and US Pharmacopoeias, in Remington's Pharmaceutical Sciences (Mack Publishing Co.) and Martindale: The Extra Pharmacopoeia (London The Pharmaceutical Press).
The disclosure of this reference is hereby incorporated into the present invention by reference. The. Pharmaceutical composition for oral administration A tablet may be prepared as follows: Component Quantity Active ingredient 10 to 500 mg Lactose 100 mg Microcrystalline cellulose 60 mg Magnesium stearate 5 Starch (eg, Amount required potato starch) to produce the total weight given below Total (per capsule) 1000 mg [0340] Alternatively, about 100 mg of a disclosed compound, 50 mg lactose (monohydrate), 50 mg cornstarch (native), 10 mg polyvinylpyrrolidone ( PVP 25) (eg from BASF, Ludwigshafen, Germany) and 2 mg of magnesium stearate are used per tablet. The active ingredient, lactose and starch mixture is granulated with a 5% w / w solution of PVP in water. After drying, the beans are mixed with magnesium stearate for 5 min. This mixture is molded using a common tablet press (eg tablet shape: 8 mm diameter, 12 mm bend radius). The applied molding force is typically about 15 kN.
Alternatively, a disclosed compound may be administered in a suspension formulated for oral use. For example, about 100 to 5000 mg of the desired disclosed compound, 1000 mg of ethanol (96%), 400 mg of xanthan gum and 99 g of water are combined with stirring. A single dose of about 10 to 500 mg of the desired disclosed compound may be provided per 10 mL of oral suspension.
In these examples, the active ingredient may be substituted with the same amount of any of the compounds according to the present invention, in particular with the same amount of any of the exemplified compounds. In some circumstances, it may be desirable to use a capsule, for example a filled gelatin capsule, rather than a tablet form. The choice of tablet or capsule will depend, in part, on the physicochemical characteristics of the particular disclosed compound used.
Examples of alternative carriers useful for making oral preparations are lactose, sucrose, starch, talc, magnesium stearate, microcrystalline cellulose, methyl cellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, carboxymethylcellulose, glycerin, sodium alginate, gum arabic, etc. Such alternative vehicles may be substituted for those indicated above as necessary for desired dissolution, absorption and manufacturing characteristics.
The amount of a compound disclosed per tablet for use in a pharmaceutical composition for human use is determined from toxicological and pharmacokinetic data obtained from appropriate animal models, for example mice, and at least one non-gnawing species, and adjusted. based on data from human clinical trials. For example, it may be appropriate for a disclosed compound to be present at a level of about 10 to 1000 mg per tablet dosage unit. B. Pharmaceutical composition for injection [0345] A parenteral composition may be prepared as follows: Component Quantity Active ingredient 10 to 500 mg Sodium carbonate 560 mg * Sodium hydroxide 80 mg * Distilled water, Amount sufficient for sterile to prepare total indicated volume below, down, beneath, underneath, downwards, downhill.
Total (per capsule) 10 mL per ampoule * Amount adjusted as needed to maintain physiological pH in the context of active ingredient amount and active ingredient form, eg a particular salt form of the active ingredient.
Alternatively, a pharmaceutical composition for intravenous injection may be used, with the composition comprising about 100 to 5000 mg of a disclosed compound, 15 g of polyethylene glycol 400 and 250 g of water in saline with optionally up to about 15%. of Cremophor EL and optionally up to 15% ethyl alcohol and optionally up to 2 equivalents of a pharmaceutically suitable acid such as citric acid or hydrochloric acid are used. The preparation of such an injectable composition may be carried out as follows: the disclosed compound and polyethylene glycol 400 are dissolved in water with stirring. The solution is subjected to sterile filtration (0.22 µm pore size) and filled into thermosilized infusion vials under aseptic conditions. Infusion vials are sealed with rubber seals.
In yet another example, a pharmaceutical composition for intravenous injection may be used, with the composition comprising about 10 to 500 mg of a disclosed compound, standard saline, optionally up to 15% by weight of Cremofor EL and optionally up to 15% by weight of ethyl alcohol and optionally up to 2 equivalents of a pharmaceutically appropriate acid such as citric acid or hydrochloric acid. The preparation may be carried out as follows: A desired disclosed compound is dissolved in saline with stirring. Optionally Cremofor EL, acid or ethyl alcohol are added. The solution is subjected to sterile filtration (0.22 µm pore size) and filled into thermosilized infusion vials under aseptic conditions. Infusion vials are sealed with rubber seals.
In this example, the active ingredient may be substituted with the same amount of any of the compounds according to the present invention, in particular with the same amount of any of the exemplified compounds.
The amount of a compound disclosed per ampoule for use in a pharmaceutical composition for human use is determined from toxicological and pharmacokinetic data obtained from appropriate animal models, for example mice, and at least one non-gnawing species, and adjusted. based on data from human clinical trials. For example, it may be appropriate for a disclosed compound to be present at a level of about 10 to 1000 mg per tablet dosage unit.
Suitable vehicles for parenteral preparations are, for example, water, saline, etc. which may be used with tris (hydroxymethyl) aminomethane, sodium carbonate, sodium hydroxide or the like, serving as a solubilizer or pH adjusting agent. Parenteral preparations preferably contain from 50 to 1000 mg of a compound disclosed per dosage unit.
It will be apparent to those skilled in the art that various modifications and variations may be made in the present disclosure without departing from the spirit and scope of the invention. Other embodiments of the invention will be apparent to those skilled in the art from the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.

权利要求:
Claims (4)
[1]
1. A compound characterized in that it has the structure represented by formula (II) wherein: m is 0 or 1; or a pharmaceutically acceptable salt thereof.
[2]
Pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in claim 1 and a pharmaceutically acceptable carrier.
[3]
Use of a compound as defined in claim 1 for preparing a pharmaceutical composition for the treatment of an uncontrolled cell proliferation disorder in a mammal.
[4]
Use of a compound as defined in claim 1 for preparing a pharmaceutical composition for reducing histone demethylase activity in a mammal.

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EP3010915A4|2016-12-28|

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KR102288648B1|2021-08-12|

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EP3010915A1|2016-04-27|

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CA2915817A1|2014-12-24|

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EP3010915B1|2019-05-08|

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JP2016523256A|2016-08-08|

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WO2014205213A1|2014-12-24|

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KR20160024929A|2016-03-07|

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法律状态:
2017-03-28| B65X| Notification of requirement for priority examination of patent application|

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2017-04-11| B65Y| Grant of priority examination of the patent application (request complies with dec. 132/06 of 20061117)|

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2017-05-16| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|

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2018-04-17| B07E| Notice of approval relating to section 229 industrial property law [chapter 7.5 patent gazette]|

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2018-10-30| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2019-01-15| B09X| Decision of grant: republication|

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2019-01-29| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 19/06/2014, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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申请号 | 申请日 | 专利标题

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