JP2017193587A

Method for preparing compositions for modulating kinase cascade, and method of using the same

Abstract

Problem to be solved.To provide a method for preparing a composition for regulating a kinase cascade and a method for using the same. The present invention relates to a composition comprising 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) --V-benzylacetamide and its mesylate and dihydrochloride. .. More specifically, the present invention presents the efficiency for the synthesis of 2- (5- <4- (2-morpholinoethoxy) phenyl) pyridin-2-yl)-^^ benzylacetamide and its mesylates and dihydrochlorides. And methods of using the compositions of the invention to regulate one or more components of the kinase cascade. [Selection diagram] None

JP2017193587A, drawing sheet 1 — Sheet 1 of 32

Term

Projected expiry 4 August 2037.

1 claim: 1 independent, 0 dependent

1
本願図面に記載された発明。

267 paragraphs, as filed

Related application This application claims priority to US Provisional Patent Application No. 60 / 930,758 filed on May 17, 2007, and is one of US Patent Application Nos. 12 / 005,792 filed on December 28, 2007. This is a continuation application. The entire contents of each patent application are incorporated herein by reference.

Field of invention The present invention relates to substantially pure 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide (KX2-391), as well as its mesylates and bis-. Concers with compositions and methods for the synthesis of hydrochlorides. The present invention also relates to a method of using such a composition.

Background of the invention Signal transduction is any process by which a cell converts one type of signal or stimulus into another. A process called signal transduction often involves a series of biochemical reactions inside the cell, which are carried out by enzymes and are related via a second messenger. In many transmission processes, increasing numbers of enzymes and other molecules become involved in events that progress from initial stimulation. In such cases, the chain of processes is referred to as the "signal transduction cascade" or "second messenger pathway" and often results in small stimuli that elicit a large response. A type of molecule involved in signal transduction is the kinase enzyme family. The largest group of kinases is protein kinases, which act on and alter the activity of specific proteins. It is widely used to transmit signals and to control complex intracellular processes.

Protein kinases are one of the larger enzyme types that catalyze the transfer of γ-phosphate of ATP to hydroxyl groups on the side chains of Ser / Thr or Tyr in proteins and peptides, and various important cells. Function, perhaps most notably: is closely involved in the regulation of signal transduction, differentiation, and proliferation. It is estimated that there are about 2,000 different protein kinases in the human body, each of which phosphorylates a particular protein / peptide substrate, but all are highly conserved of the same second substrate, ATP. Combine in the pocket. Protein phosphatase catalyzes the transfer of phosphate in the opposite direction.

Tyrosine kinases are enzymes that can convert the phosphate group of ATP to tyrosine residues in proteins. Protein phosphorylation by kinases is an important mechanism in signal transduction to regulate enzyme activity. Tyrosine kinases are divided into two groups; cytoplasmic proteins and transmembrane receptor-bound kinases. In humans, there are 32 types of cytoplasmic protein tyrosine kinases and 58 types of receptor-bound protein tyrosine kinases. Hormones and growth factors that act on cell surface tyrosine kinase binding receptors are generally growth-promoting and serve to stimulate cell division (eg, insulin, insulin-like growth factor 1, epidermal growth factor).

Various known protein kinase or protein phosphatase inhibitors have a variety of therapeutic applications. An example of a possible promising therapeutic use of a protein kinase or protein phosphatase inhibitor is its use as an anti-cancer agent. Approximately 50% of the known proto-oncogene products are protein tyrosine kinases (PTKs), the kinase activity of which has been shown to result in cellular transformation.

PTKs can be divided into two categories: membrane-receptor-type PTKs (eg, growth factor-receptor-type PTKs) and non-receptor-type PTKs (eg, the Src family of proto-oncogene products). There are at least 9 members in the Src family of non-receptor Tyrosine, pp60c-src(Hereinafter, simply referred to as "Src" in the present specification) is a basic form of PTK of the family, which is highly conserved with about 300 catalytic domains of amino acids. Overactivation of Src has been reported in several human cancers, such as colon, breast, lung, bladder, and skin cancers, as well as gastric cancer, hairy cell leukemia, and neuroblastoma. Also, hyperstimulated cell proliferation signals from transmembrane receptors (eg, EGFR and pl85HER2 / Neu) into the cell appear to pass through Src. As a result, Src has recently been proposed to be a universal target for cancer treatment because overactivation (no mutation) is involved in tumor origin, progression and metastasis in many important human tumor types. ing.

Kinases play a role in a variety of diseases and disorders because they are involved in the regulation of a wide variety of normal intracellular signaling pathways (eg, cell growth, differentiation, survival, adhesion, migration, etc.). It is believed that there is. Therefore, modulation of the kinase signaling cascade can be an important method for treating or preventing such diseases and disorders.

Recently, a small-scale synthesis of KX2-391 has been published (Patent Document 1). This synthesis is impractical for mass production of the compound and has the disadvantage that the resulting product is contaminated with ethyl chloride, which is known to be a weak alkylating agent. Therefore, the presence of sufficiently high levels of ethyl chloride limits the pharmaceutical efficacy of the KX2-391 composition.

Therefore, there is a need for an improved synthetic pathway for KX2-391 that is easy to commercialize, safe and easy, and that gives KX2-391 and its salts in large scale, high yield and substantially pure.

<patcit num="1"><text>U.S. Patent Application Publication No. 2006/01 60800</text></patcit>

Outline of the invention The compounds of the present invention are useful for modulating the components of the kinase signaling cascade. Some compounds may be useful in modulating more than one type of kinase signaling cascade. The compound of the present invention is useful as a medicinal agent. The compounds of the invention are kinases that may be involved in normal intracellular signaling pathways (eg, cell growth, differentiation, survival, adhesion, migration, etc.), or kinases that are involved in disease or disorder. Can be useful for modulation to regulate. In one aspect, the invention (1) Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to obtain 4- (2- (4-bromophenoxy) ethyl) morpholine; (2) Coupling 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-Il) Phenoxy) Ethyl) Morpholine Obtaining Step; (3) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate The process of converting to methyl; and (5) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) by reacting methyl acetate with benzylamine Step to obtain pyridine-2-yl) -N-benzylacetamide The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. In another aspect, the invention (1) Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to obtain 4- (2- (4-bromophenoxy) ethyl) morpholine; (2) Coupling 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-Il) Phenoxy) Ethyl) Morpholine Obtaining Step; (3) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate Step to convert to methyl; (5) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) by reacting methyl acetate with benzylamine Step to obtain pyridine-2-yl) -N-benzylacetamide; and (6) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide is contacted with methanesulfonic acid to make 2- (5- (4- (2-morpholino) phenyl) Step to obtain ethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate, which comprises. In another embodiment, the present invention involves contacting 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide with methanesulfonic acid to 2-(5-(5-(5-( 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate comprising the step of obtaining 4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl). )-Regarding the method for preparing N-benzylacetamide mesylate. In another embodiment, the present invention reacts methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine to react 2- (5- (4- (4- (2)) -Molholinoethoxy) Phenyl) Pyridine-2-yl) -N-Benzylacetamide, which comprises the step of obtaining 2-(5-(4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide. Regarding the preparation method of. In another embodiment, the invention presents 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine). Step to convert -2-yl) methyl acetate; and react methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine to 2- (5-(5-(5-( 4- (2-Molholinoethoxy) phenyl) Pyridine-2-yl)-Including the step of obtaining N-benzylacetamide, 2-(5-(4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl)- The present invention relates to a method for preparing N-benzylacetamide. In another embodiment, the present invention reacts 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine with acetonitrile to give 2- (5- (4- (2-morpholino) morpholino). Step to obtain ethoxy) phenyl) pyridin-2-yl) acetonitrile; 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile from 2- (5- (4- (2-) 2-yl) acetonitrile Step to convert morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate; and react methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) -N-benzylacetamide, including the step of obtaining 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) pyridine) -2-yl) -Relevant to the method for preparing N-benzylacetamide. In another embodiment, the invention couples 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (4- (4-)4-boronic acid). Step to obtain (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine; 2-(2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine Step to obtain (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile; 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile 2 -(5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) conversion to methyl acetate; and 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) ) Reacting methyl acetate with benzylamine to give 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, comprising 2- (5- (4) -(2-Molholinoethoxy) Phenyl) Pyridine-2-yl)-Relevant to the method for preparing N-benzylacetamide. In another embodiment, the invention comprises reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to give 4- (2- (4-bromophenoxy) ethyl) morpholine, 2- (5). -(4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-The method for preparing benzylacetamide. In another embodiment, the invention couples 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (4- (4-)4-) Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide, comprising the step of obtaining (6-fluoropyridine-3-yl) phenoxy) ethyl) morpholine. Regarding the method. In another embodiment, the present invention reacts 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine with acetonitrile to give 2- (5- (4- (2-morpholino) morpholino). The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises a step of obtaining ethoxy) phenyl) pyridin-2-yl) acetonitrile. In another embodiment, the present invention comprises 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) acetonitrile as 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine). -2-yl) The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises a step of converting to methyl acetate. In another embodiment, the invention uses any of the above methods for preparing KX2-391, 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl)-. Containing the step of contacting N-benzylacetamide with methanesulfonic acid to obtain 2- (5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate, 2-( The present invention relates to a method for preparing 5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate. In another aspect, the invention relates to a composition comprising KX2-391 mesylate. In another aspect, the invention relates to a composition in which KX2-391 mesylate has a purity greater than 98.0% as measured by HPLC. In another aspect, in the composition, KX2-391 mesylate has a purity of 99.0%. In another aspect, in the composition, KX2-391 mesylate has a purity of 99.5%. In another aspect, in the composition, KX2-391 mesylate has a purity of 99.6%. In another aspect, in the composition, KX2-391 mesylate has a purity of 99.7%. In another embodiment, the composition contains less than 2% impurities selected from ethyl chloride, ethanol, ethyl acetate, heptane, anisole, palladium, and combinations thereof. In another embodiment, the composition further comprises a pharmaceutically acceptable carrier or excipient. In another aspect, the invention relates to the use of a composition in the manufacture of a medicament for regulating one or more components of a protein kinase signaling cascade. In another aspect, the invention relates to the use of the composition in the manufacture of a medicament that inhibits a kinase selected from Src family protein kinases, focal adhesion kinases, and tyrosine kinases. In another embodiment, the tyrosine kinase is an Src family protein kinase. In another aspect of the invention, the medicament is orally administered. In another embodiment, the drug is administered topically. In another aspect, the invention is the use of a composition in the manufacture of a medicament for regulating one or more components of a protein kinase signaling cascade, wherein the components of the kinase cascade are hyperproliferative disorders, cancer. , Precancer, osteoporosis, cardiovascular disorders, immune system dysfunction, type II diabetes, obesity, hearing loss, and the manifestation of symptoms of the disease or disorder selected from transplant rejection. The above description fairly broadly illustrates the more important features of the invention so that the following detailed description of the invention can be understood and the contribution of the invention to the art can be better recognized. .. Other objects and features of the present invention will be self-evident from the following detailed description considered in conjunction with the examples.

<figref num="1">FIG. 1 is a graph showing the DSC of KX2-391 / 2HCl lot 02BP111F.</figref><figref num="2">FIG. 2 is a graph showing the DSC of KX2-391 / 2HCl lot 02BP111E.</figref><figref num="3">FIG. 3 is a graph showing the XRPD of KX2-391 / 2HCl lot 02BP111E.</figref><figref num="4">FIG. 4 is a graph showing the XRPD of KX2-391 / 2HCl lot 02BP111F.</figref><figref num="5">FIG. 5 is a 1H NMR spectrum of KX2-391 (lot 02BP096K).</figref><figref num="6">FIG. 6 is a 1H NMR spectrum of KX2-391 / MSA.</figref>

Detailed description of the invention Details of one or more embodiments of the present invention are given in the accompanying description below. Any method and material similar to or equivalent to that described herein may be used in the practice or testing of the present invention, but preferred methods and materials are described herein. Other features, objectives and advantages of the present invention will become apparent from the description. As used herein, the singular form also includes plurals, unless the context clearly indicates otherwise. Unless otherwise stated, all scientific and technological terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. In the case of inconsistency, it is governed by the specification of the present invention. All publications, patent applications, patents, and other references listed herein are incorporated herein by reference in their entirety.

Preparation of KX2-391 and its salts The synthesis of 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine is shown in the following scheme:

<chemistry num="1"><img id="000002" he="73" wi="109" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> 4- (2- (4- (6-Fluoripyridine-3-yl) phenoxy) ethyl) morpholine (5) was synthesized in 3 steps. Intermediate 2 was synthesized using an ether coupling reaction, for example using Williamson ether synthesis. Ether formation between 4- (2-chloroethyl) morpholine (1) and 4-bromophenol was performed in the presence of potassium carbonate and DMF to give 4- (2- (4-bromophenoxy) ethyl) morpholine (2). Obtained. Severe drying conditions were not essential for this reaction and basic washing with sodium hydroxide was used to remove residual 4-bromophenol (if any). In another aspect of the invention, Intermediate 2 is synthesized using any ether forming reaction. Intermediate 2 is synthesized starting from compound 1 containing any leaving group. For example, if you are a chemist in the art, the general formula:

<chemistry num="2"><img id="000003" he="13" wi="23" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>(In the formula, the leaving group "LG" may include, but is not limited to, a halogen (shown in compound 1), a tosyl group, a mesyl group, a triflate, etc.) as a starting material.

Compound 5 was formed using the Suzuki reaction. The formation of 6-fluoropyridin-3-yl-3-boronic acid (4), which is aryl borate, is carried out by forming an aryl anion with n-BuLi and then quenching with triisopropyl borate in place. Performed (Li et al., J. Org. Chem. 2002, 67, 5394-5397). The obtained 6-fluoropyridin-3-yl-3-boronic acid (4) was added to 4- (2- (4-bromophenoxy) ethyl) morpholine (2) in a solution of DME and aqueous sodium carbonate, and tetrakis was added. Coupling with (triphenylphosphine) palladium gives 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine (5), which is used on silica gel chromatography. Purified. A person skilled in the art will find that compound 5 is prepared using other transition metal coupling reactions.

The synthesis of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride is shown below:

<chemistry num="3"><img id="000004" he="56" wi="135" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> 2-(5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride (KX2-391HC1) was synthesized in four linear steps. The fluoride of 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine (5) was replaced with an anion of acetonitrile formed using commercially available ΝaHMDS. Slow addition of acetonitrile to a cold mixture of compound 5 and base formed 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile (6). In another aspect of the invention, Intermediate 5 may have a leaving group other than fluorine. Therefore, the compound of the general formula:

<chemistry num="4"><img id="000005" he="23" wi="52" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>(In the formula, LG includes other leaving groups known to those skilled in the art) Can be what you are trying to get.

Acid-catalyzed methanol decomposition of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile (6) was performed using a mixture of concentrated sulfuric acid and fuming sulfuric acid. The use of fuming sulfuric acid removed residual water from the reaction mixture and reduced the amount of carboxylic acid by-products formed. The reaction mixture was quenched by adding the reaction mixture to a solution of saturated sodium bicarbonate and dichloromethane, keeping the temperature below 20 ° C. Carboxylic acid contaminants (if any) were easily removed with an aqueous preparation. In another aspect of the invention, other acid-catalyzed conditions are used by those skilled in the art for the alcoholic decomposition of the nitrile of compound 6 to obtain compound 7.

The obtained 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate (7) and benzylamine were coupled in anisole at a high temperature to 2-(5-(5-( 4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide (KX2-391) was obtained. The HCl solution formed by adding acetyl chloride to absolute ethanol was added to KX2-391, bis-HCl salt, 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl). -N-Benzylacetamide dihydrochloride, (KX2-di-HCl) was formed.

The synthesis of mesylate of KX2-391 (KX2-391-MSA) is shown in the following scheme:

<chemistry num="5"><img id="000006" he="65" wi="156" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> 2-(5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate (KX2-391 / MSA) was synthesized from compound 5 for the first time in four linear steps. .. The first three steps are performed in the same manner as above for KX2-391 / 2HCl, 2-(5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate (KX2-). 391) was obtained. Treatment with methanesulfonic acid (MSA) at 50 ° C in acetone converts KX2-391 to methanesulphonate, 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl. )-N-Benzylacetamide mesylate (KX2-391, MSA) was obtained.

In another aspect of the invention, intermediate 7 having a group other than -C (O) OMe can be synthesized. If you are a chemist in the art, the general formula:

<chemistry num="6"><img id="000007" he="21" wi="59" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>(In the formula, the group "R" includes, but is not limited to, hydrogen and alkyl.) Will try to obtain intermediate compounds of.

In one aspect, the invention Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to give 4- (2- (4-bromophenoxy) ethyl) morpholine; (2) 4- (2- (4-bromophenoxy) ethyl ) Step of coupling morpholine with 6-fluoropyridin-3-yl-3-boronic acid to give 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine; 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to form 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl ) Steps to obtain acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyrididine-2-yl) acetonitrile 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine Step to convert -2-yl) methyl acetate; and (5) react methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine to 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) Step to obtain 5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises.

In another embodiment, the present invention presents (1) a step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to give 4- (2- (4-bromophenoxy) ethyl) morpholine; (2). 4- (2- (4-Bromophenoxy) ethyl) morpholine is coupled with 6-fluoropyridine-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine-3-yl) ) Step to obtain phenoxy) ethyl) morpholine; (3) 4- (2- (4- (6-fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to form 2- (5- (4- (4-) 4- (4-) Step to obtain (2-morpholinoethoxy) phenyl) pyridine-2-yl) acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) acetonitrile 2- (5) -(4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) conversion to methyl acetate; (5) 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) Step of reacting methyl acetate with benzylamine to give 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) -N-benzylacetamide; and (6) 2- (5- ( 4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) -N-benzylacetamide is contacted with methanesulfonic acid to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) )-Step to obtain N-benzylacetamide mesylate The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate, which comprises.

In another embodiment, the present invention involves contacting 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide with methanesulfonic acid to 2-(5-(5-(5-( 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate comprising the step of obtaining 4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl). )-Regarding the method for preparing N-benzylacetamide mesylate.

In another aspect, the invention (1) Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to obtain 4- (2- (4-bromophenoxy) ethyl) morpholine; (2) Coupling 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-Il) Phenoxy) Ethyl) Morpholine Obtaining Step; (3) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate Step to convert to methyl; (5) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) by reacting methyl acetate with benzylamine Step to obtain pyridine-2-yl) -N-benzylacetamide; and (6) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide is brought into contact with hydrochloric acid to make 2- (5- (4- (2-morpholinoethoxy)) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide di, including the step of obtaining phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride The present invention relates to a method for preparing a hydrochloride.

In another embodiment, the present invention involves contacting 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide with hydrochloric acid to contact 2- (5- (4- (4- (4-)4-) 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl), which comprises the step of obtaining (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride. -Regarding a method for preparing N-benzylacetamide dihydrochloride.

In another embodiment, the present invention reacts methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine to react 2- (5- (4- (4- (2)) -Molholinoethoxy) Phenyl) Pyridine-2-yl) -N-Benzylacetamide, which comprises the step of obtaining 2-(5-(4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide. Regarding the preparation method of.

In another embodiment, the invention presents 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine). Step to convert -2-yl) methyl acetate; and react methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine to 2- (5-(5-(5-( 4- (2-Molholinoethoxy) phenyl) Pyridine-2-yl)-Including the step of obtaining N-benzylacetamide, 2-(5-(4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl)- The present invention relates to a method for producing N-benzylacetamide.

In another embodiment, the present invention reacts 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine with acetonitrile to give 2- (5- (4- (2-morpholino) morpholino). Step to obtain ethoxy) phenyl) pyridin-2-yl) acetonitrile; 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile from 2- (5- (4- (2-) 2-yl) acetonitrile Step to convert morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate; and react methyl 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate with benzylamine 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) -N-benzylacetamide, including the step of obtaining 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) pyridine) -2-yl) -Relevant to the method for producing N-benzylacetamide.

In another embodiment, the present invention couples 4- (2- (4-bromophenoxy) ethyl) morpholin with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (4- (4-)4-) Step to obtain (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine; 2- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine Step to obtain (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile; 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile 2 -(5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) conversion to methyl acetate; and 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) ) Reacting methyl acetate with benzylamine to give 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, comprising 2- (5- (4) -(2-Molholinoethoxy) Phenyl) Pyridine-2-yl) -N-A method for producing benzylacetamide.

In another embodiment, the invention comprises reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to give 4- (2- (4-bromophenoxy) ethyl) morpholine, 2- (5). -(4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-The method for preparing benzylacetamide.

In another embodiment, the invention couples 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (4- (4-)4-) Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide, comprising the step of obtaining (6-fluoropyridine-3-yl) phenoxy) ethyl) morpholine. Regarding the method.

In another embodiment, the present invention reacts 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine with acetonitrile to give 2- (5- (4- (2-morpholino) morpholino). The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises a step of obtaining ethoxy) phenyl) pyridin-2-yl) acetonitrile.

In another embodiment, the present invention comprises 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) acetonitrile as 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine). -2-yl) The present invention relates to a method for preparing 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises a step of converting to methyl acetate.

In another embodiment, the present invention involves contacting 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide with methanesulfonic acid to 2- (5- (5- (5-( 2-(5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate comprising the step of obtaining 4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) )-Regarding the method for KX2-391 above for preparing N-benzylacetamide mesylate.

In another embodiment, the present invention involves contacting 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide with hydrochloric acid to contact 2- (5- (4- (4- (4-)4-) 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl), which comprises the step of obtaining (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride. -About the method for KX2-391 above for preparing N-benzylacetamide dihydrochloride.

Composition The present invention relates to substantially pure 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide (KX2-391) and its salts, solvates, Hydrate, or prodrug:

<chemistry num="7"><img id="000008" he="28" wi="95" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>Regarding. Other names for compound KX2-391 include 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide and KX2-391 free base.

The present invention relates to compositions and methods for the synthesis of high-purity KX2-391 (> 98.0% as measured by HPLC), which is safe and simple and yields KX2-391 on a large scale (> 100 g). .. Preferably, this synthesis gives the compound f in high yield (> 80%) with limited impurities.

In a preferred embodiment, KX2-391 in the compositions of the invention has a purity greater than 98%. For example, the purity of KX2-391 in the compositions of the present invention is 98.5%, 99.0%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9%.

In a preferred embodiment, the compositions and formulations of the present invention contain less than 2% impurities. For example, the compositions and formulations of the present invention contain less than 2% of any one or a combination of the following impurities: ethyl chloride, ethanol, ethyl acetate, heptane, anisole, and palladium:

Some impurities are measured in ppm, which is a relative weight measurement of solute weight / solution weight x 1,000,000, eg, ethyl chloride weight / KX2-391 diHCl sample weight x 1,000,000. For example, the weight of ethyl chloride / the weight of the KX2-391 mesylate sample x 1,000,000.

In another preferred embodiment, the composition contains 250 ppm ethyl chloride as measured by headspace gas chromatography residual solvent analysis. In one embodiment, the compounds and formulations of the present invention contain ethyl chloride in the range of about 0 ppm to about 250 ppm (or any value within the above range). For example, the composition contains less than 200 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, or less than 50 ppm of ethyl chloride.

The compounds and formulations of the present invention contain less than about 100 ppm of palladium. In one embodiment, the compounds and formulations of the present invention contain palladium in the range of about 0 ppm to about 100 ppm (or any value within the above range). For example, the composition contains less than 75 ppm, less than 50 ppm, less than 30 ppm, less than 20 ppm, less than 10 ppm, or less than 5 ppm of palladium.

In one embodiment, the compounds and formulations of the present invention contain ethanol in the range of about 0 ppm to about 5000 ppm (or any value within the above range). For example, the composition contains less than 4500 ppm, less than 4000 ppm, less than 3500 ppm, less than 3000 ppm, less than 2500 ppm, or less than 2000 ppm of ethanol.

In one embodiment, the compounds and formulations of the present invention contain ethyl acetate in the range of about 0 ppm to about 50,000 ppm (or any value within the above range). For example, the composition contains less than 48,000 ppm, less than 45,000 ppm, less than 40,000 ppm, less than 35,000 ppm, less than 30,000 ppm, or less than 25,000 ppm of ethyl acetate.

In one embodiment, the compounds and formulations of the present invention contain heptane in the range of about 0 ppm to about 7,500 ppm (or any value within the above range). For example, the composition contains less than 7,000 ppm, less than 6,500 ppm, less than 6,000 ppm, less than 5,000 ppm, less than 3,000 ppm, or less than 1,000 ppm of heptane.

In one embodiment, the compounds and formulations of the present invention contain anisole in the range of about 0 ppm to about 100 ppm (or any value within the above range). For example, the composition contains less than 80 ppm, less than 75 ppm, less than 50 ppm, less than 25 ppm, less than 10 ppm, or less than 5 ppm of anisole.

The present invention relates to compositions comprising a substantially pure solvate of KX2-391.

The present invention also relates to a composition comprising a substantially pure hydrate of KX2-391.

The present invention also comprises a substantially pure acid addition salt (eg, hydrochloride) of KX2-391. The acid addition salt can be, for example, a dihydrochloride salt. For example, the acid addition salt can be a mesylate.

The present invention relates to compositions comprising a substantially pure acid addition salt of KX2-391.

The present invention relates to a composition comprising a substantially pure hydrochloride salt of KX2-391. The present invention relates to compositions comprising substantially pure dihydrochloride of KX2-391.

The present invention relates to compositions comprising substantially pure mesylate of KX2-391.

The present invention also includes a prodrug of KX2-391.

The present invention also comprises a substantially pure pharmaceutically acceptable salt of KX2-391.

The present invention also relates to a composition comprising substantially pure KX2-391 or a solvate, hydrate or salt thereof and at least one pharmaceutically acceptable excipient.

The present invention is substantially pure 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride:

<chemistry num="8"><img id="000009" he="36" wi="84" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>Regarding.

The present invention is a composition and method for the synthesis of high-purity KX2-391 / 2HCl or KX2-391 / MSA (measured by HPLC,> 98.0%), which is safe and simple, respectively. 391.2HCl or KX2-391-MSA are obtained on a large scale (> 100 g) in high yield (> 80%) and ethyl chloride is limited (as measured by headspace gas chromatography residual solvent analysis). Regarding the method in which ethyl chloride is <250 ppm).

In a preferred embodiment, KX2-391 / 2HCl in the compositions of the invention has a purity greater than 98%. For example, the purity of KX2-391 / 2HCl in the compositions of the present invention is 98.5%, 99.0%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9%.

The present invention also relates to compositions comprising substantially pure KX2-391 / 2HCl and at least one pharmaceutically acceptable excipient.

The present invention is substantially pure 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate (KX2-391 · MSA):

<chemistry num="9"><img id="000010" he="30" wi="78" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>Regarding.

The present invention is safe and easy, KX2-391-MSA is obtained on a large scale (> 100 g) in high yield (> 80%), and ethyl chloride is limited (headspace gas chromatography residue). Concerning compositions and methods for the synthesis of high-purity KX2-391-MSA (> 98.0% as measured by HPLC) with <250 ppm ethyl chloride as measured by solvent analysis.

In a preferred embodiment, KX2-391-MSA in the compositions of the invention has a purity greater than 98%. For example, the purity of KX2-391-MSA in the compositions of the present invention is 98.5%, 99.0%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9%.

In another preferred embodiment, the composition contains less than 250 ppm ethyl chloride as measured by headspace gas chromatography residual solvent analysis. In one embodiment, the compounds and formulations of the present invention contain ethyl chloride in the range of about 0 ppm to about 250 ppm (or any value within the above range). For example, the composition contains less than 200 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, or less than 50 ppm of ethyl chloride.

The present invention also relates to a composition comprising substantially pure KX2-391-MSA and at least one pharmaceutically acceptable excipient.

Certain compounds of the invention are non-ATP competitive kinase inhibitors.

For example, the compounds of the present invention are useful for treating or preventing microbial infections, such as bacterial, fungal, parasite or viral infections.

Certain pharmaceutical compositions of the present invention contain substantially pure KX2-391 / 2HCl.

The compounds of the present invention are used as pharmaceutical agents. For example, the compounds of the present invention are used as antiproliferative agents for treating humans and / or animals, eg, for treating humans and / or other mammals. The compound can be used, for example, as an anti-cancer agent, an anti-neovascularizing agent, an anti-microbial agent, an antibacterial agent, an antifungal agent, an antiparasitic agent and / or an antiviral agent. In addition, the compounds can be used for other cell proliferation-related disorders such as diabetic retinopathy, macular degeneration and psoriasis. Examples of the anticancer agent include an antimetastasis agent.

The compounds of the invention used as pharmaceutical agents can be, for example, substantially pure KX2-391, KX2-391 / 2HCl, or KX2-391 / MSA.

The present invention provides compositions and formulations that contain a limited amount of impurities. The compounds and formulations of the present invention have a purity greater than about 98.0% when measured by methods known in the art (eg, HPLC). In one embodiment, the compounds and formulations of the present invention have a purity in the range of about 99.0% to about 100% (or any value within the above range). For example, such compounds, compositions or formulations are 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3, It can have a purity of 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%.

In order to induce the maximum pharmacodynamic and therapeutic effects of the compositions and formulations of the present invention, it is beneficial to suppress the levels of impurities such as ethyl chloride and palladium. Such impurities can result in undesired toxicity.

The compounds and formulations of the present invention contain approximately 100 ppm of palladium. In one embodiment, the compounds and formulations of the present invention contain palladium in the range of about 0 ppm to about 100 ppm (or any value within the above range). For example, the composition contains less than 75 ppm, less than 50 ppm, less than 30 ppm, less than 20 ppm, less than 10 ppm, or less than 5 ppm of palladium.

how to use Kinases play a role in a variety of diseases and disorders because they are involved in the regulation of a wide variety of normal intracellular signaling pathways (eg, cell growth, differentiation, survival, adhesion, migration, etc.). It is believed that there is. Therefore, modulation of the kinase signaling cascade can be an important method for treating or preventing such diseases and disorders. Such diseases and disorders include, for example, cancer, osteoporosis, cardiovascular disorders, immune system dysfunction, type II diabetes, obesity, and graft rejection.

The compounds of the present invention are useful for modulating the components of the kinase signaling cascade. Some compounds may be useful in modulating more than one component of the kinase signaling cascade. The phrase "regulates one or more components of a protein kinase signaling cascade" means that one or more components of a kinase signaling cascade are affected to alter cell functioning. Components of the protein kinase signaling cascade include any protein directly or indirectly involved in the kinase signaling pathway, such as a second messenger and upstream and downstream targets.

Several protein kinases and phosphatases are known and are targets for the development of therapeutic agents. For example, Hidaka and Kobayashi, Annu.Rev.Pharmacol.Toxicol, 1992, 32: 377-397; Davies et al., Biochem.J., 2000, 351: 95-105, respectively, cited herein by reference. See (embedded).

One of the kinase families, protein tyrosine kinases are two major families: receptor tyrosine kinases, or RTKs (eg, insulin receptor kinase (ERK), epidermal growth factor receptor (EGFR), basic fibroblast growth). Factor receptor (FGFR), platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR-2 or Flk1 / KDR), and nerve growth factor receptor (NGFR)) and non-receptor tyrosine kinase , Or NRTK (eg, Src family (eg Src, Fyn, Yes, Blk, Yrk, Fgr, Hck, Lck, and Lyn), Fak, Jak, Abl and Zap70). For example, Parang and Sun, Expert See Opin.Ther.Patents, 2005, 15: 1183-1207 (incorporated herein by reference).

Due to the role of Src kinases in various cancers, these kinases are the subject of several studies on the development of Src inhibitors as cancer therapeutics, such as the growth of highly metastatic cancer cells. Src inhibitors are being sought as therapeutic agents for various cancers such as colon cancer, precancerous colon lesions, ovarian cancer, breast cancer, epithelial cancer, esophageal cancer, non-small cell lung cancer, pancreatic cancer and the like. See, for example, Frame, Biochim.Biophys.Acta, 2002, 1602: 114-130 and Parang and Sun, Expert Opin.Ther.Patents, 2005, 15: 1183-1207.

Inhibition of other kinases may be useful in the treatment and modulation of other types of diseases and disorders. For example, various eye diseases can be suppressed or prevented by administration of VEGF receptor tyrosine kinase inhibitors. Inhibitors of the tyrosine phosphatase PTP-1B and / or glycogen phosphorylase may result in treatment for type II diabetes or obesity. Inhibitors of p561ck may be useful in the treatment of immune system disorders. Other targets include HIV reverse transcriptase, thromboxane synthase, EGFRTK, p55 fyn and the like.

The compounds of the invention can be Src signaling inhibitors that bind within the Src peptide substrate site. The activity of the various compounds of the invention was tested in c-Src (527F, constitutively active and transformable) transformed NIH3T3 cells and human colon cancer cells (HT29). For example, in these cell lines, KX2-391 has been shown to reduce phosphorylation levels of known Src protein substrates in a volume-dependent manner, in a volume-dependent manner, in a good correlation with growth inhibitory effects. Thus, in some embodiments, the compounds of the invention can directly inhibit Src, such as by binding to a peptide binding site (as opposed to binding at an allosteric site). possible.

Molecular model design experiments were performed to show that the compounds of the invention fit within the model Src substrate site (see, eg, US Pat. Nos. 7,005,445 and 7,070,936). Also, to target other kinases, the Src kinase inhibitor backbone using model design by simply using different sets of side chains present on the molecule and / or modifying the backbone itself. Has been renewed.

Although we do not want to be bound by theory, in cells many kinases are embedded in a multiprotein signaling complex, so that certain kinases (eg, in intracellular conformation) are compared. , Src) extracellular conformation is considered to be significantly different. Therefore, the peptide substrate binding site is not well formed in the isolated kinase (as shown by the X-ray structure of Src), so that the activity of the peptide substrate binding inhibitor on the isolated kinase is weak. Conceivable. Binding to this site in the assay of isolated kinases involves a very low proportion of total protein in the assay of the isolated enzyme in which the inhibitor is in the same conformation as the conformation present inside the cell. It is necessary to capture. Therefore, it is necessary to have a large excess of inhibitor excrete a large amount of enzyme from the catalytic cycle in the assay to make it detectable.

However, for cell line assays, a large excess of inhibitor is not needed as peptide binding sites are expected to form. In the cell-based Src assay, SH2 and SH3 domain-binding proteins have already transitioned to the Src conformation so that the peptide substrate binding site is fully formed. Therefore, since all of the enzymes are in a tightly bound conformation, even low concentrations of inhibitors can remove the enzymes from the catalytic cycle.

The overwhelming majority of known kinase inhibitors are ATP-competitive and show low selectivity in the assay panel of isolated kinases. However, many of the compounds of the present invention are considered to be peptide substrate binding inhibitors. Therefore, conventional high-performance screening of compounds against isolated enzymes such as Src cannot find the compounds of the invention.

The compound of the present invention may be a kinase inhibitor. The compounds of the present invention may be non-ATP competitive kinase inhibitors. The compounds of the present invention can directly inhibit kinases or affect kinase pathways. In one embodiment, the compound inhibits one or more components of a protein kinase signaling cascade. In another embodiment, the compound is an allosteric inhibitor. In another embodiment, the compound is a peptide substrate inhibitor. In another embodiment, the compound does not inhibit the binding of ATP to protein kinases. In one embodiment, the compound inhibits the Src family protein kinase. In another embodiment, the Src family protein kinase is pp60.c-srcIt is a tyrosine kinase.

The compounds of the present invention are useful as pharmaceutical agents, for example, as therapeutic agents for treating humans and animals. The compound can be used, for example, as an anti-cancer, anti-neoplastic, anti-metastatic, anti-microbial, anti-bacterial, anti-fungal, anti-parasitic and / or anti-viral agent.

In one embodiment, administration of the compound is topical, orally, parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, by intranasal injection, by intracavitary or intravesical injection. For example, by administering a droplet to the ear, intraarterial, intralesional, by pumping, or by applying to the mucous membrane. In another embodiment, the compound is administered with a pharmaceutically acceptable carrier.

cancer Recent literature to consider as a widely useful approach to the treatment of cancer without causing significant toxicity supports the targeting of pp60c-src (Src). For example, tumors that exhibit enhanced EGF receptor-type PTK signaling or overexpress associated Her-2 / neu receptors constitutively activate Src and enhance tumor invasiveness. Inhibition of Src in these cells causes growth arrest, induces apoptosis, and reverses the transformed phenotype (Karni et al. (1999) Oncogene). 18 (33): 4654-4662). It is known that transformed cells grow in a scaffold-independent form due to abnormally high Src activity. This was apparently that extracellular matrix signaling increased Src activity in the FAK / Src pathway, which is a coordinated form with cell division signaling, thereby activating it normally. It is caused by the fact that it blocks the fistula apoptosis mechanism. As a result, FAK / Src inhibition in tumor cells can trigger apoptosis, as it would trigger an apoptotic mechanism that would normally have begun to activate upon disruption in the absence of extracellular matrix (Hisano et al., et al. Proc.Annu.Meet.Am.Assoc.Cancer Res.38: A1925 (1997)). In addition, decreased VEGF mRNA expression was observed during Src inhibition, and tumors derived from these Src-inhibiting cell lines showed decreased angioplasty expression. (Ellis et al., Journal of Biological Chemistry 273 (2): 1052-1057 (1998)).

Src has been proposed to be a "universal" target for cancer treatment as it has been found to be overactivated in growing human tumors (Levitzki, Current Opinion in Cell Biology,). 8, 239-244 (1996); Levitzki, Anti-Cancer Drug Design, 11, 175-182 (1996)). The potential benefits of Src inhibition for cancer treatment are four-fold inhibition of uncontrolled cell proliferation caused by autocrine growth factor loop effects, inhibition of apoptosis-induced metastasis during extracellular matrix-absent disruption, VEGF. Inhibition of tumor neovascularization due to reduced levels and seems to be hypotoxic.

Prostate cancer cells have been reported to overexpress both paxillin and pl30cas and are overphosphate esterified (Tremblay et al., Int.J.Cancer, 68, 164-171, 1996). , Can be the primary target for Src inhibitors.

The present invention comprises a pharmaceutical composition comprising substantially pure KX2-391 or a salt, solvate, hydrate or prodrug thereof and at least one pharmaceutically acceptable excipient. Includes methods of preventing or treating cell proliferation disorders by administering to a subject in need of it. The present invention comprises substantially pure KX2-391 bis-HCl. The present invention comprises substantially pure KX2-391 mesylate.

For example, cell proliferation disorders are precancerous or cancerous. The cell proliferation disorders treated or prevented by the compounds of the present invention may be cancers such as colon cancer or lung cancer. The cell proliferation disorder treated or prevented by the compounds of the present invention may be an hyperproliferative disorder. The cell proliferation disorder treated or prevented by the compounds of the invention may be psoriasis.

Treatment or prevention of proliferative disorders may be by inhibition of tyrosine kinases. For example, the tyrosine kinase may be Src kinase or focal adhesion kinase (FAK).

The present invention is also a method of treating or preventing cancer or growth disorders in a subject, wherein an effective amount of substantially pure KX2-391, or a salt, solvate, hydrate or prodrug thereof. , For example, relating to a method comprising administering a composition comprising substantially pure KX2-391, KX2-391 · 2HCl or KX2-391 · MSA.

Hearing impairment As described herein, the compounds of the invention may be used to protect a subject from deafness or to prevent hearing loss in a subject. The compound may be administered to protect against deafness, before exposure to noise, or before administration of a drug that induces deafness to prevent or reduce the level of deafness. .. Drugs that induce deafness include chemotherapeutic agents (eg, platinum-based agents that target hair cells) and aminoglycoside-based antibacterial agents. The compounds of the present invention may exert a synergistic effect with a specific anticancer agent. For example, the expected inhibitors can be screened in a primary human tumor tissue assay, in particular, to find synergistic effects with other known anti-cancer agents. In addition, protein kinase inhibitors can reduce the toxicity of certain anticancer agents (eg, platinum-based agents that are toxic to the cochlea and kidneys), thereby increasing the dose.

Alternatively, the compounds of the present invention may be used to treat deafness in a subject. In this embodiment, the compound is administered to a subject after the onset of deafness to reduce the level of deafness. The compounds of the invention are involved in the regulation of the kinase cascade and are, for example, kinase inhibitors, non-ATP competitive inhibitors, tyrosine kinase inhibitors, Src inhibitors or focal adhesion kinase (FAK) modulators. Without wishing to be bound by theory, it is believed that administration of kinase inhibitors prevents apoptosis of cochlear hair cells, which prevents deafness. In one embodiment, the compounds of the invention are administered to a subject suffering from deafness to prevent further deafness. In another embodiment, the compounds of the invention are administered to a subject suffering from hearing loss in order to restore lost hearing. In particular, after exposure to noise, the close cell junctions between cochlear hair cells and the interaction of cells with the extracellular matrix are dividing and under pressure. By applying pressure to these close cell junctions, tyrosine kinases act as molecular switches, interact with focal adhesion kinases, and undergo intracellular apoptosis by a complex signaling pathway that transmits the signal of cell-matrix division to the nucleus. Start. Administration of a kinase inhibitor is thought to prevent the initiation of apoptosis in this cascade.

Confirmation of apoptosis in noise-exposed cochlear ducts has created several new possibilities for preventing noise-induced hearing loss (NIHL) (Hu et al .; 2000, Acta. Otolaryngol., 120, 19). -twenty four). For example, by administering antioxidants to the round window of the ear, the ear can be protected from NIHL (Hight et al.; 2003, Hear.Res., 179, 21-32; Hu et al.; Hear.Res.113, 198-206). Specifically, in chinchillas, NIHL was alleviated by administration of FDA-approved antioxidant compounds (NL-acetylcysteine (L-NAC) and salicylate) (Kopke et al .; 2000, Hear). .Res., 149, 138-146). In addition, Harris et al. Recently described the prevention of NIHL with Src-PTK inhibitors (Harris et al.; 2005, Hear.Res., 208, 14-25). Therefore, administration of the compounds of the invention that regulate the activity of kinases is hypothesized to be useful in treating deafness.

Changes in cell adhesion or cell stress can activate a variety of signals by integrin activation and phosphorylation of PTKs, including the Src family of tyrosine kinases. Src interactions have been linked to signaling pathways that modify the cytoskeleton and activate various protein kinase cascades that regulate cell survival and gene transcription (Giancotti and Ruoslahti; 1999, Science, 285, 1028-1032). Considered in). In fact, recent results indicate that cell-based detached outer hair cells (OHCs) underwent apoptotic cell death after exposure to intense noise. Specifically, Src The PTK signaling cascade may be involved in both metabolic and mechanically induced inhibition of apoptosis in the sensory cells of the cochlear duct. In a recent study, the Src inhibitor provided protection from noise in the 4 kHz octave band for 4 hours at 106 dB, which activates Src-PTK in outer hair cells after exposure to noise. It shows that there is a possibility (Harris et al., 2005, Hear.Res., 208, 14-25). Therefore, the compounds of the present invention that regulate the activity of Src are useful in the treatment of deafness.

Another aspect of the invention is a method of protecting a subject from deafness or treating the subject's deafness in an effective amount of substantially pure KX2-391, or a salt, solvate, water thereof. Includes methods comprising administering a solvate or prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA.

In one embodiment, the compound is administered before the onset of deafness. In another embodiment, the compound is administered after the onset of deafness.

In one embodiment, the compound is administered in combination with a deafness-causing agent, such as a cis platinum or aminoglycoside antibacterial agent. In another embodiment, the compound is administered in combination with a drug that targets hair cells.

osteoporosis The present invention relates to a method of protecting a subject from osteoporosis or treating a subject's osteoporosis. The method comprises administering to a subject an effective amount of a compound of the invention to protect or treat osteoporosis. To protect against osteoporosis, the compound may be administered before the onset of osteoporosis. Alternatively, compounds may be used to treat osteoporosis in the subject. In one embodiment, the compound is administered to the subject after the occurrence of osteoporosis in order to reduce the level of osteoporosis.

The compounds of the invention can be, for example, non-ATP competitive inhibitors. The compounds of the present invention can regulate the kinase signaling cascade depending on the particular side chain and backbone modification selected. The compounds of the present invention can be kinase inhibitors. For example, the compound can be a protein tyrosine kinase (PTK) inhibitor. High proline tyrosine kinases (PYK2; also known as cell adhesion kinase β, related adhesion-focused tyrosine kinases, or calcium-dependent tyrosine kinases), and adhesion spot kinases (FAKs) are non-receptive regulated by a variety of extracellular stimuli. Members of a different family of body protein tyrosine kinases (Avraham et al., 2000, Cell Signal, 12, 123-133; Schlaepfer et al., 1999, Prog.Biophys.Mol.Biol., 71, 435-478). The compounds of the invention can be Src inhibitors. Src deficiency has been shown to be associated with osteoporosis in mice due to loss of osteoclast function (Soriano et al., 1991, Cell, 64, 693-702). Alternatively, the compounds of the invention can regulate the expression of interleukin-1 receptor-related kinase M (IRAK-M). Mice without IRAK-M develop severe osteoporosis, which is associated with accelerated osteoclast differentiation, increased osteoclast half-life, and their activation (Hongmei et al.; 2005, J.Exp.Med., 201, 1169-1177).

Multinucleated osteoclasts arise from the fusion of mononuclear phagocytes and play a major role in bone growth and remodeling by bone reabsorption. Osteoclasts are multinucleated, terminally differentiated cells that degrade the mineralization matrix. In normal bone tissue, there is a balance between osteoblast-induced bone formation and osteoclast-induced bone reabsorption. When this dynamic and well-controlled process is out of balance, bone reabsorption can outweigh bone formation, resulting in bone loss. Osteoclasts are essential for bone development and remodeling, so increased numbers and / or activity of them can lead to diseases associated with systemic bone loss (eg, osteoporosis) and local bone loss. Other related diseases (eg, rheumatoid arthritis, periodontal disease) occur.

Both osteoclasts and osteoblasts control a number of cellular signaling pathways involved in protein kinases. Activation of osteoclasts is initiated by adhesion to bone, cytoskeletal rearrangement, formation of sealed zones, and formation of polarized wavy membranes. Protein tyrosine kinase 2 (PYK2) is thought to be involved in signal transduction from the cell surface to the cytoskeleton because it is tyrosine phosphorylated and activated by adhesion initiation signaling in osteoclasts (Duong et al., 1998, J.Clin.Invest., 102, 881-892). Recent evidence has pointed out that reduced PYK2 protein levels result in inhibition of osteoclast formation and bone reabsorption in vitro (Duong et al., 2001, J.Bio.Chem., 276, 7484-7492). Therefore, inhibition of PYK2 or other protein tyrosine kinases can reduce the level of osteoporosis by reducing osteoclast formation and bone reabsorption. Therefore, without being bound by theory, administration of the compounds of the invention regulates kinase (eg, PTK) activity and thus inhibits osteoclast formation and / or bone reabsorption, which leads to osteoporosis. Is assumed to be treated.

As confirmed in studies of Src-deficient mice and in vitro cell experiments, Src tyrosine kinases stand out as promising therapeutic targets for bone disease, with osteoclasts (positive) and osteoblasts (negative). It suggests a regulatory role for Src in both. In osteoclasts, Src plays an important role in motility, polarity, survival, activation (wavy edge formation) and adhesion, especially by mediating various signaling pathways in cytokines and integrin signaling. (Parang and Sun; 2005, Expert Opin.Ther.Patents, 15, 1183-1207). In addition, targeted disruption of the src gene in mice induces ossification, a disorder characterized by reduced bone reabsorption, without showing any overt morphological or functional abnormalities in other tissues or cells. (Soriano et al.; 1991, Cell, 64, 693-702). src-/-The osteopetrosis phenotype of mice is cell self-sustaining and is usually due to a deficiency of mature osteoclasts expressing high levels of Src protein. (Horne et al.; 1991, Cell, 119, 1003-1013). Src inhibitors are thought to reduce fractures and promote bone formation by limiting the effectiveness of Src tyrosine kinases, which induce osteoclast activity and inhibit osteoblasts. Inhibition of Src kinase activity may be useful in the treatment of osteoporosis, as osteoclasts normally express high levels of Src (Missbach et al.; 1999, Bone, 24, 437-449). Therefore, the PTK inhibitors of the present invention that regulate the activity of Src are useful in the treatment of osteoporosis.

For example, a defect in the Src gene in mice results in only one drawback: the inability to form wavy edges, resulting in osteoclasts that do not reabsorb bone. However, the bone reabsorption function of osteoclasts was rescued in these mice by inserting the kinase-deficient Src gene (Schwartzberg et al. (1997) Genes & Development 11: 2835-2844). This is clearly sufficient for the presence of the Src protein to supplement and activate other PTKs (essential for maintaining osteoclast function) in osteoclasts essential for signaling complexes. This suggests that Src kinase activity can be inhibited in vivo without inducing only known toxicity.

Another aspect of the invention is a method of protecting a subject from osteoporosis or treating a subject's osteoporosis in an effective amount of substantially pure KX2-391, or a salt, solvate, water thereof. Includes methods comprising administering a solvate or prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA.

In one embodiment, the compound is administered prior to the onset of osteoporosis. In another embodiment, the compound is administered after the onset of osteoporosis.

obesity As described herein, the compounds of the invention may be used to protect a subject from obesity or to prevent subject obesity. To protect against obesity, the compound may be administered prior to the onset of obesity in the subject. For example, the compound may be administered to prevent or reduce weight gain. Alternatively, the compound may be used to treat obesity in a subject. The compounds of the invention are involved in the modulation of the kinase signaling cascade and are, for example, kinase inhibitors, non-ATP competitive inhibitors, tyrosine kinase inhibitors, protein tyrosine phosphatase inhibitors, or protein-tyrosine phosphatase 1B inhibitors.

Obesity is often associated with diabetes and increased insulin resistance in insulin-responsive tissues such as skeletal muscle, liver and white adipose tissue (Klaman et al .; 2000, Mol.Cell.Biol, 20, 5479-5489). Insulin plays an important role in the regulation of glucose homeostasis, lipid metabolism and energy balance. Insulin signaling is initiated by the binding of insulin to the insulin receptor (IR), the receptor tyrosine kinase. Insulin binding begins with autophosphorylation of IR on multiple tyrosine residues and induces a cascade of phosphorylation events. Autophosphorylation enhances IR kinase activity and induces downstream signaling events. Most of the action of insulin is defined by the promoting action of protein tyrosine kinase and the inhibitory action of protein tyrosine phosphatase. Proper insulin signaling minimizes large fluctuations in blood glucose levels and ensures adequate delivery of glucose to cells. Since insulin stimulation causes multiple tyrosyl phosphorylation events, enhanced activity of one or more protein-tyrosine phosphatases (PTPs) can result in insulin resistance that can lead to obesity. In fact, increased PTP activity has been reported in several insulin-resistant states, including obesity (Ahmad et al .; 1997, Metabolism, 46, 1140-1145). Therefore, without wishing to be bound by theory, administration of the compounds of the invention regulates kinase (eg, PTP) activity, thereby treating obesity in a subject.

Insulin signaling begins with the activation of IR by tyrosine phosphorylation and ends with the uptake of glucose into cells by the glucose transporter GLUT4 (Saltiel and Kahn; 2001, Nature, 414, 799-806). Activated IR should then be deactivated and return to its basal state (a process in which protein-tyrosine phosphatase-1B (PTP-1B) is thought to be involved) (Ahmad et al.; 1997, J. Biol. Chem). ., 270, 20503-20508). Disruption of the gene encoding PTP-1B in mice makes them sensitive to insulin and increases resistance to dietary obesity (Elchebly et al.; 1999, Science, 283, 1544-1548; Klaman et al.; 2000, Mol. Cell.Biol, 20, 5479-5489). The decrease in fat accumulation in PTP-1B-deficient mice was due to a significant decrease in adipocyte mass without reducing adipocyte count (Klaman et al., 2000, Mol.Cell.Biol, 20, 5479-5489). ). In addition, leanness in PTP-1B-deficient mice was accompanied by an increase in basal metabolic rate and total energy expenditure without significant changes in mRNA expression of uncoupled proteins. Disruption of the PTP-1B gene has been shown to be able to regulate insulin signaling and dietary obesity in vivo by altering the activity of PTP-1B. Therefore, although not bound by theory, administration of compounds of the invention that regulate insulin signaling (eg, PTP-1B activity) is useful in the treatment of subject obesity.

Another aspect of the invention is a method of protecting a subject from obesity or treating obesity, wherein an effective amount of substantially pure KX2-391, or a salt, solvate, hydrate or hydrate thereof. Includes methods comprising administering a prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA.

In one embodiment, the compound is administered before the subject becomes obese. In another embodiment, the compound is administered after the subject becomes obese.

Diabetes As described herein, the compounds of the invention may be used to protect a subject from diabetes or to prevent diabetes. To protect against diabetes, the compound may be administered before the onset of diabetes in the subject. Alternatively, the compound may be used to treat diabetes in a subject. The compounds of the invention may be involved in the modulation of the kinase signaling cascade, eg, kinase inhibitors, non-ATP competitive inhibitors, tyrosine kinase inhibitors, phosphatase tensin homolog (PTEN) inhibitors on chromosome 10, or sequence homology. 2-Containing inositol 5'-phosphatase 2 (SHIP2) inhibitor.

Type II diabetes (T2DM) is a disorder of dysregulated energy metabolism. Energy metabolism is primarily regulated by the hormone insulin, a potent anabolic drug that promotes the synthesis and storage of proteins, carbohydrates and lipids, inhibits their breakdown and restores the circulatory system. The action of insulin is initiated by its binding to the tyrosine kinase receptor, causing autophosphorylation and increased catalytic activity of the kinase (Patti et al., 1998, J. Basic Clin. Physiol. Pharmacol. 9, 89-109). .. By tyrosine phosphorylation, the insulin receptor substrate (IRS) protein interacts with the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI3K) to activate the enzyme and move to a specific intracellular position depending on the cell type. Will result in labeling. The enzyme is a lipid product, phosphatidylinositol-3,4,5-triphosphate (PtdIns (3,4,5) P.3), Which regulates the localization and activity of numerous proteins (Kido et al.; 2001, J. Clin. Endocrinol. Metab., 86, 972-979). PI3K has important roles in insulin-stimulated glucose uptake and storage, inhibition of lipolysis, and regulation of liver gene expression (Saltiel et al.; 2001, Nature, 414, 799-806). Overexpression of the dominant interfering form of PI3K may block glucose uptake and translocation of glutamate transporter 4, GLUT4 to the cell membrane (Quon et al.; 1995, Mol.Cell.Biol, 15, 5403-5411). Therefore, administration of compounds of the invention that regulate kinase (eg, PI3K) activity and thus increase glucose uptake is useful in the treatment of diabetes.

PTEN is a major regulator of PI3K signaling in many cell types and functions as a tumor suppressor by antagonizing the anti-apoptotic, proliferative and hypertrophic activities of the PI3K pathway (Goberdhan et al.; 2003, Hum. Mol. Genet., 12, R239-R248; Leslie et al .; 2004, J. Biochem., 382, 1-11). I don't want to be bound by theory, but PTEN is PtdIns (3,4,5) P.3Dephosphorylation of the molecule attenuates the PI3K pathway, making this important lipid second messenger PtdIns (4,5) P.2It is thought that it will deteriorate. Recent studies have shown that PtdIns (3,4,5) P is due to a 50% reduction in endogenous PTEN protein using short interfering RNA (siRNA).3Increased insulin dependence and glucose uptake at levels were enhanced (Tang et al.; 2005, J. Biol. Chem., 280, 22523-22529). Therefore, although not bound by theory, administration of the compounds of the invention that regulate PTEN activity and thus result in increased glucose uptake is hypothesized to be useful in the treatment of diabetes.

Also, PtdIns (3,4,5) P3Levels are also regulated by a family of SRC homology 2 (SH2) -containing inositol 5'-phosphatase (SHIP) proteins, namely SHIP1 and SHIP2 (Lazar and Saltiel; 2006, Nature Reviews, 5, 333-342). Among other insulin-sensitive tissues, SHIP2 expressed in skeletal muscle is PtdIns (3,4,5) P.3PtdIns (3,4) P2Catalyzing the conversion to (Pesesse et al.; 1997; Biochem Biophys.Res.Commun., 239, 697-700; Backers et al.; 2003, Adv. Enzyme Regul., 43, 15-28; Chi et al.; 2004, J. et al. Biol Chem., 279, 44987-44995; Sleeman et al.; 2005, Nature Med., 11, 199-205). Insulin-stimulated PtdIns (3,4,5) P due to overexpression of SHIP23Levels are significantly reduced in harmony with the proposed volume of SHIP2, and activation of PI3K downstream effectors is diminished (Ishihara et al.; 1999, Biochem.Biophys.Res.Commun., 260, 265-272). .. Therefore, although not bound by theory, administration of the compounds of the invention that regulate SHIP2 activity and thus result in increased glucose uptake is hypothesized to be useful in the treatment of diabetes.

Another aspect of the invention is a method of protecting a subject from diabetes or treating diabetes in an effective amount of substantially pure KX2-391, or a salt, solvate, hydrate or hydrate thereof. Includes methods comprising administering a prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA.

In one embodiment, the compound is administered before the onset of diabetes. In another embodiment, the compound is administered after the disease has developed.

Eye disease As described herein, the compounds of the invention may be used to protect a subject from eye (eye) disease or to prevent eye disease. To protect against eye disease, the compound may be administered prior to the onset of eye disease in the subject. Alternatively, the compounds may be used to treat subject eye disorders such as macular degeneration, retinopathy and macular edema. The compounds of the invention are involved in the regulation of the kinase cascade, such as kinase inhibitors, non-ATP competitive inhibitors, tyrosine kinase inhibitors, such as vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitors.

New angioplasty of the physiologically avascular cornea can occur that threatens vision. Proliferative retinopathy, predominantly diabetic retinopathy and age-related macular degeneration, is characterized by increased vascular permeability, resulting in retinal edema and subretinal fluid accumulation, as well as the proliferation of new blood vessels that are prone to bleeding. Angioplasty, the formation of new blood vessels from existing capillaries, is an integral part of normal development and numerous pathological processes. VEGF, a central mediator of the complex cascade of neovascularization and an important permeability factor, is an attractive target for new therapeutic agents. VEG is a ligand for two membrane-bound tyrosine kinase receptors, VEGFR-1 and VEGFR-2. Ligand binding causes VEGFR dimerization and transphosphorylation, followed by activation of the intercellular tyrosine kinase domain. Subsequent intracellular signaling axis) causes cell proliferation, migration and survival of the vascular endothelium. Therefore, without wishing to be bound by theory, administration of compounds of the invention that regulate kinase activity, such as tyrosine kinase activity, resulting in inhibition of neovascularization and / or neovascularization. It is assumed to be useful in the treatment of eye diseases such as macular degeneration, retinopathy and / or macular edema.

Macular degeneration is characterized by VEGF-mediated retinal leakage (increased vascular permeability) and abnormal growth of small blood vessels behind the eye (new angiogenesis). VEGF has been identified in the neovascular membrane of both diabetic retinopathy and age-related macular degeneration, and the intraocular level of this factor correlates with the severity of neovascularization in diabetic retinopathy (Kvanta et al; 1996). , Invest.Ophthal.Vis.Sci, 37, 1929-1934 .; Aiello et al., 1994, N.Engl.J.Med., 331, 1480-1487). In these models, the therapeutic antagonism of VEGF results in a significant inhibition of new angioplasty in both the retina and choroid, as well as reduced vascular permeability (Aiello et al.; 1995, Proc. Natl. Acad. Sci.USA., 92, 10457-10461; Krzystolik et al.; 2002, Arch.Ophthal., 120, 338-346; Qaum et al.; 2001, Invest.Ophthal.Vis.Sci., 42, 2408-2413). Therefore, although not bound by theory, administration of the compounds of the invention that regulate VEGF activity and result in inhibition of neovascularization and / or neovascularization is associated with eye diseases such as macular edema. It is assumed to be useful in the treatment of degeneration, retinopathy and / or macular edema.

Another aspect of the invention is a method of protecting or treating a subject from eye diseases such as macular degeneration, retinopathy, macular edema, etc., in an effective amount of substantially pure KX2-391. , Or a composition comprising a salt, solvate, hydrate or prodrug thereof, eg, substantially pure KX2-391, KX2-391 / 2HC1 or KX2-391 / MSA. including.

In one embodiment, the compound is administered prior to the onset of eye disease. In another embodiment, the compound is administered after the onset of eye disease.

stroke The compounds of the present invention are used in methods of treating, preventing, or alleviating a stroke in a subject at risk of suffering a stroke, suffering a stroke, or having suffered a stroke. The compounds of the present invention are useful in methods of treating patients undergoing post-stroke rehabilitation.

Stroke, also known as a cerebrovascular attack (CVA), is an acute neurological injury that impedes the blood supply to parts of the brain by blocking arteries or rupturing blood vessels. The part of the brain where the blood supply is disrupted can no longer receive the oxygen and / or nutrients carried by the blood. Brain cells are impaired or necrotic, which impairs the function of that part of the brain or from that part. Oxygen deficiency for more than 60-90 seconds causes the brain tissue to cease functioning and, after a few minutes, suffer irreparable damage that can cause death, or infarction, of the tissue.

Stroke is classified into two main types: ischemic stroke, that is, blockade of blood vessels that supply the brain, and hemorrhagic stroke, that is, hemorrhage in or around the brain. The majority of all strokes are ischemic strokes. Ischemic stroke is generally divided into thrombotic stroke, embolic stroke, systemic hypoperfusion (diversion stroke), or venous thrombosis. In thrombotic stroke, the thrombus-forming process occurs in the affected artery, and a thrombus, or blood clot, gradually narrows the lumen of the artery, thereby blocking blood flow to the distal tissue. Such blood clots usually form around arteriosclerotic plaques. There are two types of thrombotic stroke, which are classified based on the type of blood vessel in which the thrombus is formed. Macrovascular thrombotic stroke involves the common and internal carotid arteries, the spine, and the Circle of Willis. Small vascular thrombotic stroke involves the intracerebral arteries, the branches of the Willis artery ring, the base of the middle cerebral artery, and the arteries starting from the distal vertebral and basilar arteries.

Thrombi can cause embolic stroke, which, if not occluded, becomes a thrombus at that point if the thrombus ruptures. A thrombus is a particle or fragment that travels elsewhere in the arterial bloodstream. Embolic stroke refers to the blockade of an arterial access to a part of the brain by a thrombus. Thrombi are often blood clots, but can also be plaques away from atherosclerotic blood vessels, or many other substances that can be fat, air, or cancerous cells. Since thrombosis occurs elsewhere, topical treatment only temporarily resolves the problem. Therefore, the cause of thrombus must be confirmed. There are four types of embolic stroke: known cardiac causes, potential cardiac or aortic causes (from transthoracic or transesophageal echo); arterial causes. Things; and some of unknown causes.

Systemic hypoperfusion is a decrease in blood flow to all parts of the body. This is most commonly caused by cardiac arrest or cardiac arrhythmia, or impaired cardiac output due to decreased blood excretion as a result of myocardial infarction, pulmonary embolism, pericardial effusion, or bleeding. Hypoxemia (ie, low blood oxygen content) can lead to rapid hypoperfusion. Since the decrease in blood flow is general, all parts of the brain, especially the "watershed" range, which is the border area supplied by the main cerebral arteries, are affected. Blood flow to these areas does not necessarily stop, and blood flow to the point of brain injury can be reduced.

Veins in the brain function to flush blood back to the body. When a vein is occluded due to thrombosis, blood flow is blocked, blood flows back, and cerebral edema occurs. This cerebral edema can cause ischemic and hemorrhagic strokes. This generally occurs in the bizarre disease venous sinus thrombosis.

Stroke is one or more of the various techniques known in the art, such as neurological tests, blood tests, CT scans (without contrast enhancement), MRI scans, Doppler ultrasonography and arterography (ie, no radiation). Diagnosis is made in a subject or patient using an arterial radiography) after injecting a permeable substance into the bloodstream. If the stroke is confirmed by contrast, various other tests are done to determine if there is a cause around the obstruction. Such tests include, for example, carotid artery ultrasound / Doppler tests (to detect carotid artery stenosis); electrocardiogram (ECG) and echocardiography (an arrhythmia in the heart that can spread to the cerebrovascular by blood flow and as a result). To confirm blood clots); Holter monitor examinations to confirm intermittent arrhythmia and cerebrovascular angiography (if bleeding is thought to be due to an aneurysm or arteriovenous malformation).

Compounds useful in these methods of treating, preventing or alleviating stroke or the symptoms associated with stroke are compounds that regulate the progression of the kinase signaling cascade during or after stroke. In some embodiments, the compound is a kinase inhibitor. For example, the compound is a tyrosine kinase inhibitor. In one embodiment, the tyrosine kinase inhibitor is an Src inhibitor. For example, the compounds used in the methods described herein for treating, preventing or alleviating stroke or the symptoms associated with stroke are allosteric inhibitors of the progression of the kinase signaling cascade during or after stroke. The compounds used in the stroke or methods associated with stroke described herein to treat, prevent or alleviate are preferably non-ATP competitive inhibitors of the progression of the kinase signaling cascade during or after stroke. ..

Inhibition of Src activity has been shown to protect the brain during stroke (see Paul et al., Nature Medicine, vol. 7 (2): 222-227 (2001), which is cited by reference. The whole is incorporated herein). Vascular Endothelial Growth Factor (VEGF) produced in response to ischemic injury has been shown to promote vascular permeability. Src kinase regulates VEGF-mediated VP in the brain, then stroke, and administration of Src inhibitors before and after stroke reduces edema, improves cerebral perfusion, and volume of infarct after injury. Studies have shown that is reduced (Paul et al., 2001). Therefore, Src inhibition may be useful in the treatment, treatment and alleviation of secondary injury after stroke.

The compounds of the present invention prevent, treat and alleviate the symptoms associated with stroke. Symptoms of stroke include sudden numbness or weakness, especially on one side of the body; sudden confusion, difficulty speaking, difficulty understanding conversation; sudden visual difficulty in one or both eyes; sudden difficulty walking, dizziness, or Loss of balance or coordination; or sudden, severe headache of unknown cause.

In general, stroke has three stages of treatment: prevention, immediate stroke treatment, and post-stroke rehabilitation. Treatments to prevent initial or recurrent strokes are based on treatment of the risk factors underlying the stroke, such as hypertension, high cholesterol, atrial fibrillation and diabetes. In acute stroke treatment, an attempt is made to stop the stroke by quickly dissolving the clot that causes the ischemic stroke or by stopping the bleeding of the hemorrhagic stroke. Post-stroke rehabilitation helps individuals overcome the disability caused by stroke damage. Pharmacotherapy and medication are the most common treatments for stroke. The most common types of drugs used to prevent or treat stroke are antithrombotic agents (eg, antiplatelet agents and anticoagulants) and thrombolytic agents. The compound is administered to a subject at risk of suffering a stroke, suffering a stroke, or having suffered a stroke before, during, or after a stroke, or in any combination thereof. Will be done. The compounds of the present invention may be administered alone as pharmaceutical compositions or any of a variety of known treatments such as anti-platelet drug therapy (eg, aspirin, clopidogrel, dipyridamole), anti-blood coagulants (eg, walfalin), Alternatively, it is administered in combination with thrombolytic drug therapy (eg, tissue plasminogen activator (t-PA), reteplase, urokinase, streptokinase, tenecteprase, lanoteplase, or anistreplase.

Another aspect of the invention is a method of protecting a subject from stroke or treating a stroke, wherein an effective amount of substantially pure KX2-391, or a salt, solvate, hydrate or salt thereof, or Includes methods comprising administering a prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA.

In one embodiment, the compound is administered before a stroke occurs. In another embodiment, the compound is administered after a stroke has occurred.

Atherosclerosis The compounds of the present invention are used in methods of treating, preventing or alleviating atherosclerosis or its symptoms in subjects at risk of or suffering from atherosclerosis.

Atherosclerosis is a disease that affects the blood vessels of an artery and is commonly referred to as "hardening" of the artery. Atherosclerosis results from the formation of multiple plaques in the arteries. Atherosclerotic plaques are counteracted by arterial dilation, which ultimately causes plaque rupture or narrowing of the arteries (ie, narrowing), which in turn causes the blood supply to the organs to which the blood vessels provide blood. There will be a shortage. Alternatively, excessive arterial dilation processes that are counteracted eventually result in an aneurysm. These complications are chronic, slowly progressing, and cumulative. Most commonly, soft plaques suddenly rupture, forming blood clots (ie, blood clots) that rapidly slow or stop blood flow, thereby receiving blood from the arteries. To death. This catastrophic event is called an infarction. For example, coronary thrombosis of the coronary arteries causes a myocardial infarction commonly known as a heart attack. Myocardial infarction occurs when atherosclerotic plaques slowly build up inside the coronary arteries and then suddenly rupture, blocking the entire artery and blocking downstream blood flow.

Atherosclerosis and acute myocardial infarction are diagnosed in patients using a variety of clinical and / or laboratory tests, such as physical examination, radiation or ultrasonography, and blood analysis. For example, a doctor or clinician may hear a subject's arterial sound to detect an abnormal humming sound called a brui. Bruy can be heard by placing a stethoscope over the affected artery. Alternatively or additionally, the clinician or physician can examine the pulse, for example, in the legs or feet, to find abnormalities such as weakness or lack. A physician or clinician may perform a blood test to test cholesterol levels or to test heart enzyme levels such as creatine kinase, troponin and lactate dehydrogenase to detect abnormalities. For example, the troponin subunit I or T, which is very specific for the myocardium, rises before permanent damage occurs. Positive troponin in chest pain can accurately predict that myocardial infarction is likely to occur in the near future. Other tests for diagnosing atherosclerosis and / or myocardial infarction include, for example, EKG (ECG), which measures the rate and regularity of a subject's heartbeat; a test that compares blood pressure in the ankle to blood pressure in the upper arm. Chest X-ray to measure ankle / upper arm blood pressure index; Ultrasonic analysis of arteries; CT scan of the area of interest; Angiography; Exercise stress test, nuclear heart scanning; and nuclear magnetism of the heart Resonance imaging (MRI) and positron radial tomography (PET) scans are included.

Compounds useful in these methods of treating, preventing or alleviating atherosclerosis or its symptoms are compounds that regulate the kinase signaling cascade in patients at risk or suffering from atherosclerosis. is there. In some embodiments, the compound is a kinase inhibitor. For example, the compound is a tyrosine kinase inhibitor. In one embodiment, the tyrosine kinase inhibitor is an Src inhibitor. The compounds used in the methods described herein for treating, preventing or alleviating atherosclerosis and its symptoms are preferably allosteric inhibitors of the kinase signaling cascade involved in atherosclerosis. The compounds used in the methods of treating, preventing or alleviating atherosclerosis or associated symptoms described herein are non-ATP competitive inhibitors of the kinase signaling cascade involved in atherosclerosis. Is preferable.

Intracellular signaling by Src appears to play an important role in increasing vascular permeability, known as vascular permeability (VP). For example, vascular endothelial growth factor (VEGF) produced in response to ischemic injury, including myocardial infarction, has been shown to promote vascular permeability. Studies have shown that inhibition of Src kinase reduces VEGF-mediated VP (Parang and Sun, Expert) See Opin.Ther.Patents, vol.15 (9): 1183-1206 (2005), which is incorporated herein by reference in its entirety). It has been demonstrated that mice treated with Src inhibitors have reduced tissue damage associated with trauma or damage to blood vessels after myocardial infarction (eg, US Patent Publication No. Cheresh et al.). See 20040214836 and 200303130209, these publications are incorporated herein by reference in their entirety). Therefore, inhibition of Src may be useful in the prevention, treatment or alleviation of secondary injury after injury due to atherosclerosis, such as myocardial infarction.

Atherosclerosis is generally asymptomatic until the arteries are significantly narrowed to restrict blood flow or cause a sudden occlusion. Symptoms depend on the site of plaque or stenosis, such as the heart, brain, other biological organs and legs, or any part of the body. Early symptoms of atherosclerosis are when the body needs more oxygen, for example during exercise, when a person feels chest pain due to a lack of oxygen in the heart (angina), or to the legs. It can be pain or spasm when feeling leg cramps due to lack of oxygen. Narrowing of the arteries that supply blood to the brain can cause dizziness or transient ischemic attack (TIA), in which case the symptoms or signs of stroke last for less than 24 hours. Typically, these symptoms progress gradually.

Symptoms of myocardial infarction are characterized by varying degrees of chest pain, discomfort, sweating, weakness, chills, vomiting, and arrhythmias, and can also cause loss of consciousness. Chest pain is the most common symptom of acute myocardial infarction and is often described as tension, tightness, and squeezing. Pain may spread to the chin, neck, arms, back and epigastrium, most often to the left arm or neck. Chest pain is more likely to occur if myocardial infarction lasts longer than 30 minutes. Patients with myocardial infarction may exhibit shortness of breath (dyspnea), especially if the decrease in myocardial contractility due to the infarction is sufficient to cause left ventricular insufficiency with embryonic congestion or even embryonic edema.

The compounds of the invention are administered alone as pharmaceutical compositions or for the treatment of any variety of known atherosclerosis, such as cholesterol-lowering agents (eg, statins), antiplatelet drug therapy, or anticoagulants. Is administered in combination with.

Another aspect of the invention is a method of protecting a subject from atherosclerosis or treating atherosclerosis in an effective amount of substantially pure KX2-391, or a salt thereof. Includes methods comprising administering a solvate, hydrate or prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA. In one embodiment, the compound is administered before the onset of symptoms of atherosclerosis. In another embodiment, the compound is administered after the onset of symptoms of atherosclerosis.

Neuropathic pain The compounds of the present invention treat neuropathic pain, such as chronic neuropathic pain, or its symptoms in subjects at risk of, suffering from, or have suffering from neuropathic pain. Used in preventative or mitigating methods.

Neuropathic pain, also known as neuralgia, is qualitatively different from normal nociceptive pain. Neuropathic pain is usually present as a constant burning and / or "tingling" and / or "electric shock" -like sensation. The difference between nociceptive pain and neuropathic pain, for the "normal" nociceptive pain is to stimulate only the pain nerve, neuropathic pain, Shiba Shiba, pain sensations in the same range Due to the fact that it stimulates both nerves and non-painful sensory nerves (eg, nerves that respond to tactile, warm, and cold sensations), resulting in the generation of signals that the spinal cord and brain would not normally receive. ..

Neuropathic pain is usually a complex chronic pain condition with tissue damage. For neuropathic pain, the nerve fibers themselves can be damaged, dysfunctional, or injured. These damaged nerve fibers send false signals to other pain centers. The effects of nerve fiber injury include changes in nerve function both at the site of injury and in the area surrounding the injury.

Neuropathic pain is diagnosed in a subject or patient using one or more different laboratory and / or clinical techniques known in the art, such as a medical examination.

Compounds useful in treating, preventing or alleviating neuropathic pain, such as chronic neuropathic pain, or the symptoms associated with neuropathic pain, are compounds that regulate the kinase signaling cascade involved in neuropathic pain. Is.

c-Src has been shown to regulate the activity of the N-methyl-D-aspartate (NMDA) receptor (Yu et al., Proc.Natl.Acad.Sci.USA, vol.96: 7697- See 7704 (1999), which is incorporated herein by reference in its entirety). Studies have shown that PP2, a low molecular weight Src kinase inhibitor, reduces phosphorylation of the NMDA receptor NM2 subunit (Guo et al., J. Neuro., Vol. 22: 6208-6217 (2002). ) Reference, the document is incorporated herein by reference in its entirety). Therefore, Src inhibition, which eventually inhibits the activity of NMDA receptors, may be useful in the prevention, treatment or alleviation of neuropathic pain such as chronic neuropathic pain.

The compounds of the present invention prevent, treat or alleviate neuropathic pain, such as chronic neuropathic pain, or the symptoms associated with neuropathic pain. Symptoms of neuropathic pain include stinging pain, burning pain, tingling and numbness.

The compounds of the present invention may be administered alone as pharmaceutical compositions or may be any of a variety of known therapies such as analgesics, opioids, tricyclic antidepressants, anticonvulsants and serotonin norepinephrine reuptake inhibitors. It is administered in combination with antidepressant.

In one embodiment, the compound is administered prior to the onset of chronic neuropathic pain. In another embodiment, the compound is administered after the onset of chronic neuropathic pain.

Hepatitis B The compounds of the present invention are used in methods of treating, preventing or alleviating hepatitis B or its symptoms in subjects at risk or suffering from hepatitis B.

Hepatitis B virus, a member of the hepadnavirus family, has a protein core particle containing a viral genome in the form of double-stranded DNA with a single-stranded region and an outer lipid-based envelope with an embedded protein. It is composed of. Encapsular proteins are involved in viral binding and release into susceptible cells. The internal capsid rearranges the DNA genome into the nucleus of the cell, where the viral mRNA is transcribed. Along with the transcript encoding the X protein, three subgenome transcripts encoding the envelope protein are created. A fourth pregenomic RNA is transcribed, which is transported to the cytosol to translate viral polymerases and core proteins. Polymerase and pregenomic RNA are capsidated in the aggregation of core particles, where reverse transcription of the pregenomic RNA into genomic DNA occurs by the polymerase protein. Mature core particles then exit the cell through the normal secretory pathway, but acquire the envelope along the way.

Hepatitis B is one of several known nonretroviral viruses that employ reverse transcription as part of the replication process. Other viruses that use reverse transcription include, for example, HTLV or HIV.

During HBV infection, the host's immune response causes both hepatocyte damage and viral clearance. Congenital immune responses do not play an important role in these processes, but adaptive immune responses, especially virus-specific cytotoxic T lymphocytes (CTLs), are responsible for almost all causes of liver damage associated with HBV infection. Become. CTL also eliminates the virus by producing antiviral cytokines that can kill infected cells and expel HBV from living hepatocytes. Liver damage is initiated and mediated by CTL, but non-antigen-specific inflammatory cells can exacerbate CTL-induced immunopathology, and platelets can promote the accumulation of CTL in the liver.

Hepatitis B is diagnosed in patients using any variety of clinical and / or laboratory tests, such as health examinations, and blood or serum analysis. For example, blood or serum is tested for the presence of viral antigens and / or antibodies produced by the host. In general hepatitis B trials, detection of hepatitis B surface antigens (HBsAg) is used to screen for the presence of infection. This is the first detectable viral antigen that appears during this viral infection, but because this antigen is cleared by the host, it may not be present in the early stages of infection and is undetectable in the later stages of infection. It may be possible. During this "window" where the host remains infected but is successfully eliminating the virus, IgM antibodies (anti-HBc IGM) against hepatitis B core antigens may be the only serological evidence of the disease.

Immediately after the appearance of HBsAg, another antigen named hepatitis B e-antigen (HBeAg) appears. Traditionally, the presence of HBeAg in host serum has been implicated in a very high rate of virus replication, but some hepatitis B virus variants do not produce any "e" antigens. There are also things. During the natural course of infection, HBeAg is eliminated and antibodies against the "e" antigen (anti-HBe) are produced shortly thereafter. This conversion is usually associated with a dramatic reduction in virus replication. If the host can eliminate the infection, then HBsAg will eventually become undetectable, followed by antibodies to the hepatitis B surface antigen (anti-HB). Those who are negative for HBsAg but positive for anti-HB are those who have ruled out the infection or have been previously vaccinated. Many people who are positive for HBsAg have little viral growth and may therefore have little risk of long-term complications or infection of others.

Compounds useful in these methods of treating, preventing, or alleviating hepatitis B or its symptoms are compounds that regulate the kinase signaling cascade in patients at risk or suffering from hepatitis B. In some embodiments, the compound is a kinase inhibitor. For example, the compound is a tyrosine kinase inhibitor. In one embodiment, the tyrosine kinase inhibitor is an Src inhibitor. The compounds used in the methods described herein for treating, preventing or alleviating hepatitis B or its symptoms are preferably allosteric inhibitors of the kinase signaling cascade involved in hepatitis B. The compounds used in the methods described herein for treating, preventing, or alleviating hepatitis B or the symptoms associated with hepatitis B are non-ATP competitive inhibitors of the kinase signaling cascade involved in hepatitis B. It is preferable to have.

Src plays a role in the replication of hepatitis B virus. The virus-encoded transcription factor HBx activates Src in the steps required by the transmission of the HBV virus (eg, Klein et al., EMBO J., vol. 18: 5019-5027 (1999); Klein et al. , Mol.Cell.Biol., Vol.17: 6427-6436 (1997), each of which is incorporated herein by reference in its entirety). Therefore, Src inhibition, which in turn inhibits Src-mediated transmission of the HBV virus, may be useful in the treatment, treatment or alleviation of hepatitis B or its symptoms.

The compounds of the present invention prevent hepatitis B or symptoms associated with hepatitis B. Treat or alleviate. Symptoms of hepatitis B typically develop within 30-180 days after exposure to the virus. However, up to half of all people infected with the hepatitis B virus are asymptomatic. Symptoms of hepatitis B are often compared to influenza, such as loss of appetite; fatigue; nausea and vomiting, generalized itching; liver pain (eg, right side of the abdomen, under the lower thorax), jaundice, And changes in excretory function.

The compounds of the invention are administered alone as pharmaceutical compositions or in combination with any of a variety of known hepatitis B treatments such as interferon α, lamivudine (Epivir-HBV) and Baraclude (entecavir).

Another aspect of the invention is a method of protecting a subject from hepatitis B or treating hepatitis B in an effective amount of substantially pure KX2-391, or a salt, solvate thereof. Includes methods comprising administering a hydrate or prodrug, eg, a composition comprising substantially pure KX2-391, KX2-391 / 2HCl or KX2-391 / MSA.

In one embodiment, the compound is administered before the subject is infected with hepatitis B. In another embodiment, the compound is administered after the subject has been infected with hepatitis B.

Control of immune system activity As described herein, the compounds of the invention control the activity of the subject's immune system, thereby controlling autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, sepsis and lupus, and implants. It may be used to protect against rejection and allergic diseases, or to prevent them. Alternatively, the compound may be used to treat an autoimmune disease of the subject. For example, the compound results in a reduction in the severity of symptoms or an imminent cessation of progression of the subject's autoimmune disease. The compounds of the invention are involved in the modulation of the kinase signaling cascade and are, for example, kinase inhibitors, non-ATP competitive inhibitors, tyrosine kinase inhibitors, such as Src inhibitors, p59fyn (Fyn) inhibitors or p561ck (Lck) inhibitors.

Autoimmune diseases are diseases caused by disruption of self-tolerance, such as the adaptive immune system responding to self-antigens and mediating cell and tissue damage. Autoimmune diseases can be organ-specific (eg, thyroiditis or diabetes) or systemic (eg, systemic lupus erythematosus). T cells regulate the cell-mediated immune response in the adaptive immune system. Under normal conditions, T cells express antigen receptors (T cell receptors) that recognize peptide fragments of heterologous proteins bound to automajor histocompatibility complex molecules. The earliest perceived events after stimulation with the T cell receptor (TCR) include activation of Lck and Fyn, which are TCR residues on tyrosine residues within the immunoreceptor tyrosine system activation motif. (Zamoyska et al., 2003, Immunol. Rev., 191, 107-118). Tyrosine kinases such as Lck (a member of the Src family of protein tyrosine kinases) play an important role in cell signaling and modulation of cell proliferation by phosphorylating peptide and protein tyrosine residues (Levitzki; 2001). , Top.Curr.Chem., 211, 1-15; Longati et al .; 2001, Curr.Drug Targets, 2, 41-55; Qian and Weiss; 1997, Curr.Opin.Cell Biol, 9, 205-211). Therefore, although not bound by theory, administration of compounds of the invention that regulate tyrosine kinase (eg, Src) activity is hypothesized to be useful in the treatment of autoimmune diseases.

Both tyrosine kinases lck and fyn are activated in the TCR pathway, so inhibitors of lck and / or fyn have potential utility as autoimmune agents (Palacios and Weiss; 2004, Oncogene, 23, 7990). -8000). Lck and Fyn are predominantly expressed by T cells throughout their lifetime. The role of Lck and Fyn in T cell development, homeostasis and activation has been demonstrated by animal and cell line studies (Parang and Sun; 2005, Expert). Opin.The.Patents, 15, 1183-1207). Lck activation is involved in autoimmune disease and graft rejection (Kamens et al .; 2001, Curr. Opin. Investig. Drugs, 2, 1213-1219). The result is that the lck (-) jarcat cell line proliferates, produces cytokines, and increases intracellular calcium, inositol phosphate, and tyrosine phosphorylation in response to stimulation of T cell receptors. Indicates that it cannot be done (Straus and Weiss; 1992, Cell., 70, 585-593; Yamasaki et al.; 1996, Mol. Cell.Biol, 16, 7151-7160). Therefore, drugs that inhibit lck effectively block T cell function, act as immunosuppressants, autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and lupus, and graft rejection and allergies. It has potential utility in the area of sexual disorders (Hanke and Pollok; 1995, Inflammation Res., 44, 357-371). Therefore, without being bound by theory, administration of compounds of the invention that regulate one or more members of the Src family of protein tyrosine kinases (eg, lck and / or fyn) is a treatment for autoimmune diseases. It is assumed to be useful in.

Another aspect of the invention is a method of controlling the activity of the immune system of a subject, wherein an effective amount of substantially pure KX2-391, or a salt, solvate, hydrate or prodrug thereof. For example, a method comprising administering a composition comprising substantially pure KX2-391, KX2-391 · 2HCl or KX2-391 · MSA.

Definition For convenience, the specific terms used in this specification, examples and the accompanying claims are collected here.

Protein kinases catalyze the transfer of γ-phosphate from ATP to the hydroxyl groups on the side chains of Ser / Thr or Tyr in proteins and peptides, and various important cellular functions (probably the most notable signal). It is a large type of enzyme that is closely involved in the regulation of transmission, differentiation and proliferation. There are about 2,000 different protein kinases in the human body, each phosphorylating a particular protein / peptide substrate, all of which bind to the same secondary substrate ATP in a highly conserved pocket. It is estimated that. About 50% of the known proto-oncogene products are protein tyrosine kinases (PTKs), and their kinase activity has been shown to result in cellular transformation.

PTK can be divided into two categories: membrane receptor PTK (eg, growth factor receptor PTK) and non-receptor PTK (eg, Src family of proto-oncogene products and focal adhesion kinase (FAK)). Can be done. Overactivation of Src has been reported in a number of human cancers, including cancers of the colon, breast, lung, bladder and skin, as well as in gastric cancer, hairy cell leukemia and neuroblastoma.

"Inhibiting one or more components of a protein kinase signaling cascade" means implementing one or more components of a kinase signaling cascade to alter cell function. Components of the protein kinase signaling cascade include second messengers and any protein that is directly or indirectly involved in the kinase signaling pathway, including upstream and downstream targets.

"Treatment" includes any effect that results in improvement of the condition, disease, disorder, etc., such as relieving, reducing, adjusting, or eliminating. As a "treatment" of a disease state or a "treatment" of a disease state, inhibiting the disease state, that is, stopping the progression of the disease state or its clinical symptoms, or alleviating the disease state, that is, the disease state. Or it may cause a temporary or permanent regression of its clinical symptoms.

"Preventing" a disease state is a condition of the disease in a subject who has been or is prone to the disease state but has not yet experienced or does not have symptoms of the disease state. Includes preventing the onset of clinical symptoms.

"Disease state" means any disease, disorder, condition, symptom, or sign.

As used herein, the term "cell proliferation disorders" refers to conditions that can be cancerous or non-cancerous due to uncontrolled and / or abnormal cell proliferation, such as unwanted conditions or diseases such as psoriasis. The condition that can cause the occurrence of. As used herein, the term "psoriasis" or "psoriasis" refers to disorders including keratin cell hyperproliferation, inflammatory cell infiltration, and cytokine degeneration.

In one embodiment, the cell proliferation disorder is cancer. As used herein, the term "cancer" refers to solid tumors such as lung, breast, colon, ovary, brain, liver, pancreas, prostate cancer, malignant melanoma, non-melanoma skin cancer, and childhood leukemia and lymphoma. Hematological tumors and / or malignancies such as, multiple myeloma, Hodgkin's disease, lymphoma caused by lymphocytes and skin, acute and chronic leukemia such as acute lymphoblastic, acute myelogenous or chronic myelogenous leukemia, traits Included are cell tumors, lymphoid tumors and cancers associated with AIDS.

In addition to psoriasis, epidemic and dermoid cysts, lipomas, adenomas, capillaries and cutaneous hemangiomas, lymphangioma, as types of proliferative disorders that can be treated using the compositions of the invention , Nevus lesions, teratomas, renal tumors, myofibromatosis, osteogenic tumors, and other dysplastic masses. Proliferative disorders include dysplasia and disorders.

"Therapeutically effective amount" means an amount sufficient to carry out the treatment of the disease when the compound is administered to the mammal for the treatment of the disease. In one embodiment, a therapeutically effective amount is administered to the mammal to reduce the level of the disease, eg, the level of deafness. In one embodiment, a therapeutically effective amount of the compound is administered. In another embodiment, a therapeutically effective amount of the composition is administered. The "therapeutically effective amount" varies depending on the compound, the disease and its severity, the age of the mammal to be treated, the body weight, and the like.

A therapeutically effective amount of one or more compounds can be formulated with a pharmaceutically acceptable carrier for administration to humans or animals. Thus, a compound or formulation can be administered, for example, by oral, parenteral or topical route to provide a therapeutically effective amount of the compound. In an alternative embodiment, the compounds prepared according to the present invention can be used to coat or impregnate medical devices such as stents.

The term "preventive effective amount" means an effective amount of a compound (s) of the present invention that is administered to cause disease prevention. In one embodiment, a prophylactically effective amount of the compound is administered. In another embodiment, a prophylactically effective amount of the composition is administered.

As used herein, "pharmacological effect" includes an effect produced in a subject that achieves an intended therapeutic purpose. In one embodiment, the pharmacological effect means that the main symptom of the subject being treated is prevented, alleviated, or reduced. For example, the pharmacological effect is to provide prevention, alleviation, or reduction of the main sign in the subject to be treated. In another embodiment, the pharmacological effect means that the symptoms of the disorder or major sign of the subject being treated are prevented, alleviated or reduced. For example, the pharmacological effect is to result in prevention or reduction of the main symptom in the subject to be treated.

The compounds of the present invention containing nitrogen are converted to N-oxides by treatment with an oxidizing agent (eg, 3-chloroperoxybenzoic acid (m-CPBA) and / or hydrogen peroxide), and the other of the present invention. Compounds can be produced. Therefore, all nitrogen-containing compounds shown and claimed are, where valence and structure allow, the indicated compound and its N-oxide derivative (N O or N).+-O-Can be shown as). In addition, in other cases, the nitrogen of the compounds of the invention can also be converted to N-hydroxy or N-alkoxy compounds. For example, N-hydroxy compounds can be prepared by oxidizing the parent amine with an oxidizing agent such as m-CPBA. All nitrogen-containing compounds shown and claimed are the compounds shown and their N-hydroxy (eg N-OH) and N-alkoxy (eg N-OR (eg N-OR)) where the valence and structure allow. In the formula, R is substituted or unsubstituted C1-6Alkyl, C1-6Alkenyl, C1-6Alkyne, C3-14It is considered to include both carbon rings or 3- to 14-membered heterocycles)) derivatives.

"Counterions" are used to represent small, negatively charged species such as chlorides, bromides, hydroxides, acetates and sulfates.

As used herein, "anionic group" refers to a group that is negatively charged at physiological pH. Anion groups include carboxylates, sulfates, sulfonates, sulfinates, sulfamates, tetrazolyl, phosphates, phosphonates, phosphinates or thiophosphates, or their functional equivalents. Can be mentioned. A "functional equivalent" of an anionic group includes a bioisostere, eg, a bioisostere of a carboxylate group. Bioisosteres include classical biobiological equivalents and non-classical bioisosteres. Classical and non-classical bioisosteres are known in the art (eg, Silverman, RBThe Organic Chemistry of Drug Design and Drug Action, Academic Press, Inc .: San Diego, Calif., 1992, pp. See .19-23). In one embodiment, the anionic group is carboxylate.

The present invention includes all isotopes of atoms present in the compound. Isotopes contain atoms that have the same atomic number but different mass numbers. Examples of hydrogen isotopes include, but are not limited to, tritium and deuterium, and carbon isotopes include C-13 and C-14.

Some of the compounds described herein have an asymmetric center. Compounds of the invention containing asymmetrically substituted atoms can be isolated in optically active or racemic form. For example, methods of producing an optically active form by dividing the racemic form or synthesizing it from an optically active starting material are well known in the art. Many geometric isomers of olefins, such as C = N double bonds, may also be present in the compounds described herein, and all such stable isomers are intended herein. Is to be done. The cis and trans geometric isomers of the compounds of the invention can be described and isolated as a mixture of isomers or in the form of separated isomers. All chiral, diastereomeric, racemic and geometric isomer forms of the structure are intended unless a particular stereochemical or isomer form is specifically indicated. All tautomers of the compounds shown or described are also considered part of the present invention.

In the present specification, for convenience in some cases, the structural formulas of compounds represent specific isomers, but the present invention presents with structurally occurring isomers such as geometric isomers, asymmetric carbon-based optical isomers, sterics It includes all isomers such as isomers, isomers, etc., and isomer mixtures, and is not limited to the description of the formula for convenience, and may be any one or mixture of isomers. Therefore, the asymmetric carbon atom may be present in the molecule, and an optically active compound and a racemic compound may be present in the present compound, but the present invention is not limited thereto and is arbitrary. Including those. In addition, polymorphs of crystals can also exist, but are not limited, and any crystalline form can be alone, a mixture of crystalline forms, or anhydrous or hydrated. Moreover, so-called metabolites produced by in vivo degradation of the compound are also included within the scope of the invention.

"Heterosexual" means compounds that have the same molecular formula but differ in properties, or the order in which their atoms are bonded, or in the arrangement of their atoms in space. Isomers with different arrangements of their atoms in space are called "stereoisomers". Three-dimensional isomers that are not mirror images of each other are called "diastereoisomers", and three-dimensional isomers that are non-overlapping enantiomers are called "enantiomers" or are sometimes called optical isomers. The carbon atom attached to four non-identical substituents is called the "chiral center".

"Chiral isomer" means a compound having at least one chiral center. The chiral isomer has two enantiomeric forms of opposite chirality and can exist as individual enantiomers or as a mixture of enantiomers. A mixture containing the individual enantiomeric forms with the same amount of reverse chirality is referred to as a "racemic mixture". Compounds with multiple chiral centers are 2n-1It has an enantiomer pair, where n is the number of chiral centers. Compounds with multiple chiral centers can exist as individual diastereomers or as a mixture of diastereomers called a "diastereomeric mixture". If one chiral center is present, the stereoisomer may be characterized by an absolute configuration (R or S) of that chiral center. Absolute configuration refers to the spatial configuration of substituents attached to the chiral center. Substituents attached to the stereocenter under consideration are ranked according to the Cahn, Ingold and Prelog ranking rules (Cahn et al., Angew.Chem.Inter.Edit.1966, 5,385; errata511; Cahn et al., Angew .Chem.1966, 78, 413; Cahn and Ingold, J.Chem.Soc.1951 (London), 612; Cahn et al., Expertia 1956, 12, 81; Cahn, J., Chem.Educ.1964, 41, 116 ).

"Geometric isomer" means a diastereomer that is present by the binding rotation of a double bond. Such arrangements follow the Cahn-Ingold-Prelog rules and are named by the prefixes cis and trans, or Z and E, indicating that the groups are on the same or opposite sides of the double bond in the molecule. Distinguished by.

In addition, the structures and other compounds discussed herein include all of their atropisomers. An "atropisomer" is a type of steric isomer in which two isomer atoms are arranged spatially differently. Atropisomers are present by the restricted rotation caused by the binding rotation of large groups around the central bond. Such atropisomers usually exist as mixtures, but as a result of recent advances in chromatographic techniques, it is possible to separate mixtures of two atropisomers when selected.

The term "crystal polymorph" or "polymorph" or "crystal morphology" means a crystal structure in which a compound (or a salt or solvate thereof) can all crystallize in different crystal packed arrangements with the same elemental composition. .. Different crystal forms usually have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability and solubility. Depending on the recrystallization solvent, crystallization rate, storage temperature and other factors, one type of crystal morphology may predominate. Crystal polymorphs of compounds can be produced by crystallization under different conditions.

Furthermore, the compounds of the invention, such as salts of the compounds, can also exist in hydrated or non-hydrated (anhydride) forms or as solvates with other solvent molecules. Non-limiting examples of hydrates include monohydrates, dihydrates and the like. Non-limiting examples of solvates include ethanol solvates, acetone solvates and the like.

"Solvate" means a solvent addition form comprising a stoichiometric or non-stoichiometric amount of solvent. Some compounds have a tendency to confine solvent molecules of constant molar ratio in the crystalline solid state, thereby forming a solvate. If the solvent is water, the solvate formed is a hydrate, and if the solvent is an alcohol, the solvate formed is alcoholate. A hydrate is formed by combining one or more water molecules and one substance, and in the hydrate, water has a molecular state of H.2Keeping as O, such a combination can form one or more hydrates.

"Tautomer" refers to a compound that exists in a simple and quick equilibrium state, although its structure differs significantly in the arrangement of atoms. It should be understood that the compounds of the invention can be presented as different tautomers. It is also understood that when a compound has a tautomeric form, all tautomeric forms are included within the scope of the present invention and the naming of the compound does not exclude any tautomeric form. Should be done.

Some compounds of the invention may exist in tautomeric form. Tautomers are also intended to be included within the scope of the present invention.

The compounds, salts and prodrugs of the invention can be present in several tautomeric forms, including enol and imine forms, keto and enamine forms, and geometric isomers and mixtures thereof. All such tautomeric morphologies are within the scope of the present invention. The tautomer is present in solution as a mixture of a set of tautomers. In the solid form, one tautomer is usually predominant. The present invention includes all tautomers of the compound, even if only one tautomer is described.

A tautomer is one of two or more structural isomers that exist in equilibrium and are easily converted from one isomer form to another. This reaction causes the formal transfer of hydrogen atoms at the same time as the switching of adjacent conjugated double bonds. In a solution capable of tautomerization, a chemical equilibrium of the tautomer is achieved. The exact ratio of tautomers depends on several factors, including temperature, solvent and pH. The concept of tautomers that can be converted to each other by tautomerization is called tautomerism.

Of the various types of tautomerization possible, two are commonly observed. In keto-enol tangle, electrons and hydrogen atoms migrate at the same time. Ring-chain tautomerism is indicated by glucose. Ring-chain tautomerization occurs as a result of reacting an aldehyde group (-CHO) in a sugar chain molecule with a hydroxy group (-OH) in the same molecule, resulting in a ring (cyclic) form.

Tautomerization is base: 1. deprotonation; 2. formation of delocalized anions (eg, enolate); 3. protonation at different positions of anions; acid: 1. protonation; 2. non-protonation. Formation of localized cations; 3. Catalyzed by deprotonation at different positions adjacent to the cations.

Common tautomer pairs are ketone-enol, amide-nitrile, lactam-lactim, amide-imidic acid tautomers in heterocycles (eg, in the nucleic acid bases guanidine, timine and cytosine), amines. -Enamine and Enamine-Enamine.

Therefore, unless otherwise suggested, isomers resulting from asymmetric carbon atoms (eg, all enantiomers and diastereomers) should be understood to be within the scope of the present invention. Such isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis. In addition, the structures, other compounds and moieties discussed herein also include all of their tautomers. Alkenes may include either E or Z geometry, where appropriate. The compounds of the present invention may exist in the form of stereoisomers and can therefore be prepared as individual stereoisomers or mixtures.

A "pharmaceutical composition" is a preparation containing the disclosed compound in a form suitable for administration to a subject. In one embodiment, the pharmaceutical composition is in bulk or unit dosage form. Formulating the composition into a unit dosage form may be advantageous in terms of ease of administration and uniform dosage. As used herein, a unit dosage form refers to a physically individual unit suitable as a unit dose for a subject to be treated, each unit together with the required pharmaceutical carrier. Contains a predetermined amount of active reagent calculated to produce the desired therapeutic effect. Specifications for unit dosage forms of the invention are dictated by the unique characteristics of the active reagent and the particular therapeutic effect to be achieved, as well as the discipline-specific limitations of formulating such active agents for personal treatment. And depend directly on them.

The unit dosage form may be any of a variety of forms, including, for example, capsules, IV bags, tablets, single spray doses with a spray inhaler, or vials. The amount of the active ingredient (eg, the disclosed compound or a salt, hydrate, solvate or isomer formulation thereof) in a unit dose of the composition is an effective amount and varies depending on the specific treatment involved. .. Those skilled in the art will appreciate that depending on the age and condition of the patient, it may be necessary to constantly change the dose. The dose also depends on the route of administration. Various routes are intended, such as oral, lung, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal mucosa, sublingual, intrapleural, subspider, intranasal administration, etc. Can be mentioned. Dosage forms for topical or transdermal administration of the compounds of the invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. In one embodiment, the active compound is aseptically mixed with a pharmaceutically acceptable carrier and any preservative, buffer, or propellant required.

The term "flash dose" refers to a compound formulation that quickly disperses a dosage form.

The term "immediate release" is defined as the release of a compound from a dosage form in a relatively short time, generally up to about 60 minutes. The term "controlled release" is defined to include delayed release, sustained release and pulsed release. The term "pulse release" is defined as the successive release of a drug from a dosage form. The term "sustained release" or "sustained release" is defined as the long-term continuous release of a compound from a dosage form.

"Subjects" include mammals such as humans, pet animals (eg dogs, cats, birds, etc.), livestock (eg cows, sheep, pigs, horses, chickens, etc.) and laboratory animals (eg, rats, etc.). Mice, guinea pigs, birds, etc.). In one embodiment, the subject is human.

As used herein, the phrase "pharmaceutically acceptable" is valid within reasonable medical scope without causing excessive toxicity, irritation, allergic response or other problems or complications. A compound, substance, composition, carrier and / or dosage form suitable for use in contact with human and animal tissues in a profit / risk ratio.

"Pharmaceutically acceptable excipients" are generally safe, non-toxic, and useful excipients for the production of pharmaceutical compositions that are neither biologically nor otherwise desirable. Means, and examples thereof include excipients that are acceptable for veterinary and human pharmaceutical applications. As used herein and in the claims, "pharmaceutically acceptable excipients" include one or more of such excipients.

The compounds of the present invention can further form salts. Also, all of these forms are included within the scope of the claimed invention.

"Pharmaceutically acceptable salt" of a compound means a salt that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound.

As used herein, "pharmaceutically acceptable salt" is a derivative of the disclosed compound, wherein the parent compound is modified by producing an acid or base salt thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, minerals or organic acid salts of basic residues such as amines, alkalis or organic salts of acidic residues such as carboxylic acids. .. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts or quaternary ammonium salts of parent compounds formed from non-toxic inorganic or organic acids. For example, as such conventional non-toxic salts, 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonic acid, citric acid, edetic acid, ethanedisulfone. Acids, 1,2-ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, glycollyarsanilic acid, hexylresorcinic, hydrabamic acid, hydrobromic acid , Hydrochloride, hydroiodic acid, hydroxymaleic acid, hydroxynaphthoic acid, isetionic acid, lactic acid, lactobionic acid, laurylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, napcilic acid (napsylic), nitrate, shu Acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, stearic acid, basic acetic acid (subacetic), succinic acid, sulfamic acid, sulfanic acid, sulfuric acid, tannic acid, Examples include, but are not limited to, tartaric acid, toluenesulfonic acid, and salts derived from inorganic and organic acids selected from commonly obtained amine acids such as glycine, alanine, phenylalanine, arginine and the like.

Other examples include hexanoic acid, cyclopentanepropionic acid, pyruvate, malonic acid, 3- (4-hydroxybenzoyl) benzoic acid, silicic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfon. Acids, Drosophila sulfonic acid, 4-methylbicyclo- [2.2.2] -oct-2-en-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, muconic acid and the like can be mentioned. Further, in the present invention, when the acidic protons present in the parent compound are replaced with metal ions such as alkali metal ions, alkaline earth ions, or ammonium ions, or ethanolamine, diethanolamine, triethanolamine, tromethamine, It also includes salts that are formed when coordinated with organic bases such as N-methylglucamine.

It should be understood that all references to pharmaceutically acceptable salts also include the solvent-added form (solvate) or crystalline form (polymorph) of the salt as defined herein.

The pharmaceutically acceptable salt of the present invention can be synthesized from a parent compound containing a basic or acidic moiety by a conventional chemical method. In general, such salts are made by reacting the free acid or base form of these compounds with a chemical amount of the appropriate base or acid in water or an organic solvent or a mixture thereof. And non-aqueous solvents such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile can be used. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, Volume 18 (Mack Publishing Company, 1990). For example, examples of the salt include, but are not limited to, the aliphatic amine-containing compound of the present invention, the hydroxylamine-containing compound, and the hydrochloride and acetate of the imine-containing compound.

The compounds of the present invention can be prepared as prodrugs, eg, pharmaceutically acceptable prodrugs. The terms "pro-drug" and "prodrug" are used herein interchangeably to refer to any compound that releases an active parent drug in vivo. Prodrugs are known to enhance a number of desirable properties of pharmaceuticals (eg, solubility, bioavailability, manufacturability, etc.) so that the compounds of the invention can be delivered in prodrug form. Therefore, the present invention is intended to include a prodrug of the compound claimed in the present invention, a method for delivering the prodrug, and a composition containing the prodrug. The "prodrug" is intended to include any covalently bound carrier that releases the active parent drug of the invention in vivo when the prodrug is administered to a subject. The prodrug of the present invention is produced by modifying a functional group present in a compound by a normal operation or a method in which the modified product is cleaved into the parent compound in vivo. As a prodrug, a hydroxy, amino, sulfhydryl, carboxy or carbonyl group is attached to any group, which cleaves in vivo and is a free hydroxyl group, a free amino group, a free sulfhydryl group, a free carboxy group or a free carbonyl group, respectively. Examples of the compounds of the present invention are such that they form.

Examples of prodrugs are hydroxy functional group esters (eg, acetate, dialkylaminoacetate, formic acid ester, phosphate ester, sulfate ester, and benzoate ester derivatives) and carbamates (eg, N, N-dimethyl). Aminocarbonyl), ester groups of carboxyl functional groups (eg ethyl esters, morpholinoethanol esters), N-acyl derivatives (eg N-acetyl), N-Mannich bases, Schiff bases and enaminones of amino functional groups, Examples include, but are not limited to, oximes, acetals, ketals, and enol esters of ketone and aldehyde functional groups in the compounds. See Bundegaard, H. "Design of Prodrugs," p1-92, Elesevier, New York-Oxford (1985).

"Protecting group" refers to a group of atoms that mask, reduce or suppress reactivity when attached to a reactive group in a molecule. Examples of protecting groups are Green and Wuts, Protective Groups in Organic Chemistry, (Wiley, 2nd Edition, 1991); Harrison and Harrison et al., Compendium of Synthetic Organic Methods, Volumes 1-8 (John Wiley and Sons, 1971-). It can be found in 1996); and Kocienski, Protecting Groups (Verlag, 3rd edition, 2003).

"Stable compound" and "stable structure" are meant to indicate a compound that is robust enough to withstand the isolation of the reaction mixture to a useful degree of purity and the formulation into an effective therapeutic agent. ..

As used herein, the singular form also includes the plural form unless the context clearly dictates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the event of any inconsistency, this specification will prevail.

All percentages and percentages used herein are weight-based, unless otherwise indicated.

A "combination therapy" (or "co-treatment") is a particular treatment regimen in which a compound of the invention and at least a second agent are intended to have a beneficial effect from the joint action of these therapeutic agents. Including administration as part. Beneficial effects of the combination include, but are not limited to, the pharmacokinetic or pharmacodynamic interactions caused by the combination of therapeutic agents. Administration of these combinations of therapeutic agents is usually performed for a defined time (usually minutes, hours, days or weeks, depending on the combination selected). "Combination therapy" may also include administration of two or more of these therapeutic agents, although not commonly, as part of a separate monotherapy regimen that happens to occur accidentally and optionally in combination with the present invention. ..

"Combination therapy" is a continuous method of administering these therapeutic agents, that is, administering each therapeutic agent at a different time and substantially simultaneously administering these therapeutic agents or at least two therapeutic agents. It includes administration in. Substantial co-administration can be achieved, for example, by administering to the subject a single capsule containing each therapeutic agent in a fixed proportion, or a plurality of capsules for each therapeutic agent. Continuous or substantially simultaneous administration of each therapeutic agent is by any suitable route, such as, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal tissue. It can be carried out. Therapeutic agents can be administered by the same or different routes. For example, the first therapeutic agent of the selected combination may be administered by intravenous injection and the other therapeutic agent of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered by intravenous injection. The order in which the therapeutic agents are administered is not very strict.

"Combination therapy" also includes administering the therapeutic agents described above in combination with other biologically active ingredients and non-drug treatments (eg, surgery or radiation treatment). If the combination therapy further comprises a non-drug treatment, the non-drug treatment can be performed at any suitable time as long as a beneficial effect is achieved from the combined action of the therapeutic agent and the non-drug treatment. .. For example, when appropriate, even when non-drug treatment is temporarily, perhaps for days or even weeks, withdrawn from administration of the therapeutic agent, beneficial effects are still achieved.

Throughout the disclosure, where a composition is described as having, containing or containing a characteristic component, the composition is essentially composed of or consisting of the listed components. Is also intended. Similarly, if a method is described as having, containing or containing a particular treatment step, the method shall consist essentially of the listed treatment steps or consist of the listed treatment steps. Is also intended. Furthermore, it should be understood that the order of steps or the order in which certain actions are performed is not important if the invention is feasible. Furthermore, two or more steps or actions may be performed simultaneously.

The compound, or a pharmaceutically acceptable salt thereof, can be taken orally, nasally, transdermally, transpulmonary, inhaled, transbuccomucosally, sublingually, intraperitoneally. It is administered subcutaneously, intramuscularly, intravenously, intrarectally, intrapleurally, sublingually, and parenterally. In one embodiment, the compound is administered orally. Those skilled in the art will recognize the benefits of a particular route of administration.

Dosage regimens that utilize compounds include the type, species, age, weight, gender and medical condition of the patient; the severity of the condition to be treated; the route of administration; the patient's renal and liver function; and the specific compound used. Or it is selected according to various factors such as its salt. A physician or veterinarian with conventional skills can easily determine and prescribe the effective amount of drug required to stop the progression of the condition and to counterattack or stop it.

The disclosed techniques for the formulation and administration of the compounds of the present invention can be found in Remington: the Science and Practice of Pharmacy, 19th Edition, Mack Publishing Co., Easton, PA (1995). In one embodiment, the compounds described herein, and pharmaceutically acceptable salts thereof, are used in pharmaceutical preparations in combination with pharmaceutically acceptable carriers or diluents. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents, and sterile aqueous or organic solutions. The compound is present in the pharmaceutical composition in an amount sufficient to provide the desired dose within the range described herein.

In one embodiment, the compound is prepared for oral administration, the compound disclosed herein or a salt thereof is mixed with a suitable solid or liquid carrier or diluent and capsules, tablets, pills, powders, syrups, solutions or suspensions. Form a turbid liquid or the like.

Tablets, pills, capsules, etc. are about 1 to about 99% by weight of active ingredient and binders like tragant gum, acacia, corn starch or gelatin; excipients like dicalcium phosphate; corn starch, potato starch Or disintegrants such as alginic acid; including lubricants such as magnesium stearate and / or sweeteners such as saccharin, lactose, saccharin, xylitol and the like. When the unit dosage form is a capsule, it often contains a liquid carrier such as fatty oil in addition to the above-mentioned types of substances.

In some embodiments, various other substances are present as a coating or to denature the physical form of a dose unit. For example, in some embodiments, the tablets are coated with shellac, sugar, or both. In some embodiments, the syrup or elixir includes, in addition to the active ingredient, saccharose as a sweetener, methylparaben and propylparaben as preservatives, colorants, and flavoring agents such as cherry or orange flavors.

For some embodiments relating to parenteral administration, the disclosed compounds, or salts, solvates, tautomers or polymorphs thereof, combine with sterile aqueous or organic media to form injectable solutions or suspensions. can do. In one embodiment, the injectable composition is an aqueous isotonic solution or suspension. The composition may be sterilized and / or include an adjuvant such as a preservative, stabilizer, wetting or emulsifying agent, a solubilizing agent, a salt and / or a buffer that controls osmotic pressure. In addition, it may contain other therapeutically beneficial substances. Each composition is prepared according to conventional mixing, granulation or coating methods and contains about 0.1-75% active ingredient, and in another embodiment the composition contains about 1-50% active ingredient. Including.

For example, an injectable solution is prepared using a solvent such as sesame oil or peanut oil, an aqueous propylene glycol solution, and an aqueous solution of a water-soluble, pharmaceutically acceptable salt of the compound. In some embodiments, the suspension is made in glycerol in oil, liquid polyethylene glycol and mixtures thereof. Under normal storage and use conditions, these preparations contain preservatives to block the growth of microorganisms. As used herein, the terms "parenteral administration" and "parenteral administration" refer to routes other than enteral and topical administration, usually by injection, intravenously and muscularly. Intramuscular, intraarterial, subspider, intraarticular, intraorbital, intracardiac, intradermal, intraperitoneal, enteral, subcutaneous, subepithelial, intraarticular, subcapsular, subspider, intraspinal and intrathoracic injections and infusions However, it is not limited to these.

For intrarectal administration, suitable pharmaceutical compositions are, for example, topical preparations, suppositories or enemas. The suppository is advantageously prepared from a fat emulsion or suspension. The composition may contain sterilized and / or adjuvants such as preservatives, stabilizers, wetting agents or emulsifiers, solubilizers, salts and / or buffers that control osmotic pressure. In addition, it may contain other therapeutically beneficial substances. Each composition is prepared according to conventional mixing, granulation or coating methods and contains about 0.1-75% active ingredient, and in another embodiment the composition contains about 1-50% active ingredient. Including.

In some embodiments, the compound is such that the active agent is delivered by pulmonary administration, eg, by administration of an aerosol containing the active agent, selected from a manual pump atomizer, nebulizer or pressurized metered dose inhaler. It is formulated. In some embodiments, suitable formulations of this type also include other agents, such as antistatic agents, to keep the disclosed compounds as effective aerosols.

The drug delivery device for delivering the aerosol comprises a suitable aerosol container with a measurement valve, including the described pharmaceutical aerosol formulation, and an actuator housing provided to support the canister and enable drug delivery. Including. A canister in a drug delivery device has a headspace of more than about 15% of the total capacity of the canister. Polymers for pulmonary administration are often dissolved, suspended or emulsified in a mixture of solvents, surfactants and propellants. The mixture is maintained under pressure in a canister sealed with a measuring valve.

For nasal administration, either solid or liquid carriers can be used. The solid support comprises a coarse powder having a particle size in the range of, for example, about 20 to about 500 microns, and the formulation is administered by rapid aspiration through the nostrils. In some embodiments where a liquid carrier is used, the formulation is administered as a nasal spray or droplet and comprises an oil solution or aqueous solution of the active ingredient.

Active reagents can be prepared using carriers that protect against rapid disappearance from the body. For example, controlled release formulations can be used, including implants and microencapsulated delivery systems. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoester and polylactic acid can be used. How to prepare such a formulation will be apparent to those skilled in the art. The substance is also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells as well as monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. It can be prepared according to methods known to those of skill in the art, such as those described in US Pat. No. 4,522,811.

In addition, the compositions and formulations of the present invention may also contain one or more desiccants. Suitable desiccants that can be used in the present invention are pharmaceutically safe and include, for example, pharmaceutical grade silica gel, crystalline sodium, potassium or calcium aluminosilicate, colloidal silica, anhydrous calcium sulfate and the like. Be done. The desiccant may be present in an amount of about 1.0% to 20.0%, or about 2% to 15% w / w (or any value within the above range).

Also intended is a formulation that is a rapidly dispersing dosage form, also known as a "flash dose" form. In particular, some embodiments of the invention allow their active ingredients to be applied for a short period of time, eg, less than about 5 minutes, in another embodiment less than about 90 seconds, in another embodiment about 30 seconds. It is formulated as a composition that releases in less than a second, and in another embodiment, in less than about 10 or 15 seconds. Such formulations are suitable for administration to a subject by a variety of routes, eg, by inserting into a body cavity or by applying to a moist body surface or open wound.

Usually, a "flash dosage form" is a solid dosage form that is orally administered, which disperses quickly in the mouth and therefore does not require much effort to swallow and the compound is quickly ingested or absorbed through the oral mucosa. Make it possible. In some embodiments, suitable rapidly dispersing dosage forms include the treatment of wounds and other physical injuries and disease states for which the drug cannot be released by externally supplied water. It is also used for the purpose of.

"Flashdose" forms are known in the art and are, for example, effervescent dosage forms of US Pat. Nos. 5,578,322 and 5,607,697 and rapid release coatings of insoluble microparticles; US Pat. Nos. 4,642,903 and 5,631,023. Freeze-dried foams and liquids; melt spinning of US Pat. Nos. 4,855,326, 5,380,473 and 5,518,730 dosage forms; solid-free form production of US Pat. No. 6,471,992; US Pat. Nos. 5,587,172, 5,616,344, 6,277,406. And 5,622,719, Saccharide Carrier Matrix and Liquid Binding Agents; and other forms known in the art.

The compounds of the present invention are also formulated as "pulse-releasing" formulations in which the compounds are sequentially (i.e., pulsed) released from the pharmaceutical composition. The compound is also formulated as a "sustained release" formulation in which the compound is continuously released from the pharmaceutical composition over an extended period of time.

Also, cyclic or acyclic encapsulating agents, or solvators, such as cyclodextrins, polyethers or polysaccharides (eg, methylcellulose), or, in other embodiments, alkyl ether spacer groups or polysaccharides for lipophilic vacancies. Formulations such as liquid formulations containing a polyanionic β-cyclodextrin derivative with a sodium sulfonic acid base separated from the are also intended. In one embodiment, the encapsulating agent or solvating agent is methylcellulose. In another embodiment, the encapsulant or solvate is a polyanionic β-cyclodextrin derivative with sodium sulfonate separated from lipophilic vacancies by a butyl ether spacer group, eg, CAPTISOL® (CyDex). , Overland, KS). A person skilled in the art will prepare an aqueous solution of the agent, eg, a 40 wt% solution, prepare a serial dilution, eg, 20%, 10, 5% of the solution, for the proper ratio of the agent / disclosed compound formulation. Add excess of the disclosed compound (compared to the amount in which the drug can be solubilized), such as 2.5%, 0% (control); under appropriate conditions, eg, under heating, stirring, ultrasonic treatment. It can be determined by mixing with, etc., centrifuging or filtering the resulting mixture to obtain a clear solution, and analyzing the solution with respect to the concentration of the disclosed compound.

All publications and patent documents cited herein are incorporated herein by citation, as each of these publications or documents has been shown to be incorporated herein by citation specifically and individually. Be incorporated. Citations of publications and patent documents do not acknowledge to be relevant prior documents, nor do they give any permission in terms of content or date. With respect to the inventions described so far by the specification, those skilled in the art can implement the invention in various embodiments, the above description and the following examples are for purposes of explanation and are described below. You will understand that it is not intended to limit the claims that follow.

Example Example 1: Small-scale synthesis of KX2-391<chemistry num="10"><img id="000011" he="29" wi="82" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> The pre-synthesis described below is that shown in US20060160800A1. This procedure is useful for small-scale reactions, eg, reactions that yield up to 50 g of product. In the following synthesis, reagents and solvents were used as received from commercial sources, unless otherwise stated. Proton and carbon nuclear magnetic resonance spectra were acquired on a Bruker AC 300 or Bruker AV 300 spectrometer at 300 MHz for protons and 75 MHz for carbon. The spectrum is shown in units per ppm (δ) and the coupling constant J is reported in units of hertz. Tetramethylsilane was used as the internal standard for the proton spectrum and the solvent peak was used as the reference peak for the carbon spectrum. Mass spectra and LC-MS mass data were obtained on a Perkin Elmer Sciex 100 atmospheric ionization (APCI) mass spectrometer. LC-MS analysis value is Luna Obtained using a standard solvent gradient program (Method B) by UV detection at 254 nm using a C8 (2) Column (100 × 4.6 mm, Phenomenex). Thin layer chromatography (TLC) was performed using an Analtech silica gel plate and visualized with ultraviolet (UV) light, iodine, or 20 wt% phosphomolybolic acid-containing ethanol. HPLC analysis values were obtained using a Prevail C18 column (53 × 7 mm, Alltech) with UV detection at 254 nm using a standard solvent gradient program (Method A or B).<tables num="2"><img id="000012" he="76" wi="76" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></tables> Synthesis of N-benzyl-2- (5-bromopyridin-2-yl) acetamide:<chemistry num="11"><img id="000013" he="22" wi="50" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> In a flask, 5- (5-bromopyridin-2 (1H) -iriden) -2,2-dimethyl-1,3-dioxane-4,6-dione (1.039 g, 3.46 mmol), benzylamine (0.50 mL, 4.58 mmol) and toluene (20 mL) were charged. The reaction was refluxed under nitrogen for 18 hours, then cooled and placed in a freezer until cold. The product was collected by filtration and washed with hexane to give a clear-colored crystalline mass (1.018 g, 96%). Synthesis of 4- (2-(4- (4,4,5,5-tetramethyl [1,3,2] dioxaborolan-2-yl) -phenoxy) ethyl) morpholine:<chemistry num="12"><img id="000014" he="24" wi="54" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> 4- (4,4,5,5-tetramethyl [1,3,2] dioxaborolan-2-yl) -phenol (2.55 g, 11.58 mmol), 2-morpholin-4-ylethanol (1.60 mL, 1.73 g) , 13.2 mmol) and triphenylphosphine (3.64 g, 13.9 mmol) in a stirred solution of methylene chloride (60 mL) at 0 ° C. DIAD (2.82 g, 13.9 mmol) was added dropwise. The reaction mixture was heated to room temperature and stirred overnight. After 18 hours, some additional triphenylphosphine (1.51 g, 5.8 mmol), 2-morpholine-4-ylethanol (0.70 mL, 5.8 mmol), and DIAD (1.17 g, 5.8 mmol) were added. After stirring at room temperature for an additional 2 hours, the reaction is concentrated and the residue is flash chromatographed (CHCl).3Purification with medium 5% -25% EtOAc) gave the product as a white solid (2.855 g, 74%). 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide KX2-391 synthesis<chemistry num="13"><img id="000015" he="30" wi="81" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> N-benzyl-2- (5-bromopyridin-2-yl) acetamide (123 mg, 0.403 mmol), 4- (2- (4- (4- ( 4,4,5,5-tetramethyl [1,3,2] dioxaborolan-2-yl) -phenoxy) ethyl) morpholine (171 mg, 0.513 mmol), and FiberCat 10071(30 mg, 0.015 mmol) was charged. After adding ethanol (3 mL), an aqueous potassium carbonate solution (0.60 mL, 1.0 M, 0.60 mmol) was added. The tubes were sealed and heated at 150 ° C. for 10 minutes under microwave conditions. The reaction was cooled and concentrated to remove most of the ethanol, then dissolved in 10 mL of ethyl acetate and washed continuously with water and saturated sodium chloride solution. EDTA the organic layer4It was dried in, filtered and concentrated to give a white solid. This white solid was ground with ethyl ether to give KX2-391 as a white solid (137 mg, 79%): mp 135-137 ° C;11 H NMR (300MHz, CDCl3) δ 8.70 (d, 1H, J = 2.0Hz), 7.81 (dd, 1H, J = 2.4Hz, J = 8.0Hz), 7.65 (br s, 1H), 7.49 (d, 2H, J = 8.8Hz) , 7.37-7.20 (m, 6H), 7.01 (d, 2H, J = 8.8Hz), 4.49 (d, 2H, J = 5.8Hz), 4.16 (t, 2H, J = 5.7Hz, 3.82 (s, 2H) ), 3.78-3.72 (m, 4H), 2.84 (t, 2H, J = 5.7Hz), 2.62-2.58 (m, 4H); HPLC (Method B) 98.0% (AUC), tR= 1.834 minutes; APCI MS m / z 432 [M + H]+。1 Polymer-bonded di (acetate) dicyclohexylphenylphosphine palladium (II), available from Johnson Matthey, Aldrich (catalog number 590231). Example 2: Medium-scale synthesis of KX2-391 dihydrochloride The synthesis outlined in this example can be used in medium-scale reactions. The preparation of a batch of at least 50 g of KX2-391 dihydrochloride is shown in Scheme 1. This linear synthesis consisted of six steps, the seventh step was the preparation of one of the reagents, 6-fluoropyridin-3-ylboronic acid, which is also commercially available. The overall yield of this sequence was 35%, the average yield was 83%, and the lowest yield step was 68%. Of these seven steps, only one step required chromatography. The procedure described below was performed on a 70 g scale.<chemistry num="14"><img id="000016" he="124" wi="160" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> The first step is K2CO3Williamson ether synthesis between 4-bromophenol (131 g) using acetonitrile as a solvent and N-chloroethylmorpholine (1; 141 g as an HCl salt) using powder (3 to 3.5 equivalents) as a base. The ingredients were mixed and stirred under reflux overnight to a high degree of conversion (96.3-99.1%). After diluting with dichloromethane and heptane, the reaction mixture was filtered and evaporated to give the desired product 2 in essentially quantitative yield (216 g). Note that similar substrates (eg 4-bromo-3-fluorophenols), conversions (even with further heating), are not always high (eg 59.9-98.3%). Alkane chloride and K2CO3Both were preferably purchased from Aldrich. Continuous heating does not accelerate the reaction to terminate, and unreacted bromophenol can be easily removed by dissolving the crude reaction mixture in 4 parts of toluene and rinsing the phenol with 4 parts of 15% NaOH aqueous solution. .. One of the reagents required for the second step (Suzuki coupling) was 6-fluoropyridin-3-ylboronic acid (4). Although commercially available, this reagent is 5-bromo-2-fluoropyridine (3,102 g) in n-butyllithium (1.2 eq) at low temperature (<-60 ° C) in TBME. It was readily prepared by the addition of triisopropyl borate (1.65 eq) after lithium bromide replacement. Both stages of this reaction are short and the total reaction time (including addition time) is about 3 hours. Quenching is performed with 24% aqueous NaOH, which also extracts the product and leaves impurities in the organic layer. After removing the aqueous layer, it is then neutralized with HCl and extracted with EtOAc. After the organic layer has dried, it is diluted with a small amount of heptane and concentrated to result in precipitation / crystallization of the product. Filtration gave boronic acid 4 in relatively high purity (96.4% AUC) and good yield (69 g, 79-90%; see notes on yield estimation in the experimental section). It can be used without further purification. The second reaction step (Suzuki coupling) of this linear sequence is a reaction that is easy to set up; all reagents [2 (111 g), aqueous Na2CO3, DME, and Pd (PPh)3)4(0.04 eq)] was placed in a reaction flask and the mixture was heated under reflux; note that the reaction mixture was degassed to remove oxygen. When the reaction is complete (within 7 hours), the operation involves decanting (or sucking up) the reaction solution from the organic salts on the sides of the flask (no visible aqueous layer), rinsing and drying the flask. , The solvent was removed from the combined organic layer. Crystallization of crude 5 from isopropanol / heptane gave a material with improved purity compared to crude, but still required chromatography (silica gel ratio to crude was about 8.5: 1). The material was of sufficient purity (> 98%); the yield was 68% (79.5 g). The use of transparent 5 avoided the need for the next step, chromatography with acetonitrile substitution of fluorine atoms. Also, the exchange of fluoride with acetonitrile was a simple reaction, and clear crystallization of the crude product at room temperature gave transparent 6 in high yield and high purity. The reaction is first accompanied by the addition of 5 (79 g) of fluoride immediately after the formation of "enolate" from acetonitrile (6.5 eq) with hexamethyldisilane potassium KHMDS (8 eq) / THF at -10 ° C. It was a thing. The reaction was rapid and after 1 hour, quenching was performed with saturated brine. After drying the organic layer and evaporating the solvent, the resulting crude mixture consisted of only two components: the desired product and the apparently much less polar product due to self-condensation of acetonitrile. It was. The crude mixture was swirled in isopropanol / heptane and allowed to stand overnight, which resulted in complete crystallization of the product, which was filtered off and washed to a good high purity 6 (99.3% AUC). It was obtained in a high yield (64 g, 76%). Decompose 6 (64 g) of methanol with 40% H until the reaction is complete (25 hours).2SO4This was done by heating in (in MeOH). Then, the reaction solution is cooled and DDL is used.4Stir with and trace amounts of hydrolysis products (ArCH)2-CO2Me) converted back to product and then cooled aqueous K2CO3And at the same time extracted in dichloromethane. After drying and evaporation of most DCMs, the addition of 5% EtOAc (in heptane) and further concentration resulted in product crystallization. Filtration and washing of this solid gave a high purity (98.9% AUC) of 7 in good yield (82%) and a high purity product (4 g) from the mother liquor in a total yield of 61.7 g (87%). ). In addition, the amidation step involved charging various components (7 (61 g), benzylamine (3 equivalents), and high boiling anisole) into the reaction vessel, and then heating under reflux until the reaction was completed. .. Cooling of the reaction mixture resulted in complete crystallization of the target compound in high purity (98.9%) and good yield (81%). The final step was the formation of the dihydrochloride of the target compound. To ensure complete protonation at both basic sites, the reaction was carried out in anhydrous ethanol, which is well soluble in the dihydrochloride. After near evaporation to dryness, the reaction mixture was "followed" twice with ethanol to remove excess hydrogen chloride. The resulting viscous oil was dissolved in ethanol (2 parts) and then added to a large volume (20 parts) of EtOAc (ethyl acetate) with high speed stirring. Filtration, washing with ethyl acetate (without heptane) and vacuum drying gave KX2-391 dihydrochloride as a milky white powder. The final result was a total of 68 g (97% yield) of salt in high purity (99.6% AUC), which contained traces of EtOAc (4.8% w / w), EtOH (0.3% w / w), and heptane. (0.6% w / w; from final wash with heptane before vacuum drying) was included. The salt was also crystallized from high temperature EtOH / EtOAc (instead of the precipitation method described above) to give crystalline beads. It had much lower capture solvent levels (only 0.26% w / w EtOAc and 0.45% w / w EtOH) and was free-flowing.<chemistry num="15"><img id="000017" he="29" wi="107" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 4- (2- (4-Bromophenoxy) Ethyl) Morpholine (2): 1 (140.7g, 0.756mol), 4-bromophenol in a 5L 3-neck round-bottom flask equipped with an automatic stirrer, a thermometer with an adapter, a concentrator, and a nitrogen supply port (top of the concentrator) (130.6g, 0.755mol), anhydrous K2CO3Powder (367.6 g, 2.66 mol, 3.5 eq) and acetonitrile (1.3 L) were charged. The mixture was vigorously stirred at 80 ° C. (overnight) (blade contacting the bottom of the flask), then diluted with DCM (500 mL) and heptane (200 mL) and filtered through Celite. Evaporation to dryness (rotational concentration followed by high vacuum) gave 2 as a pale yellow oil (216.00 g, 100% yield, 96.3% AUC, 3.7% unreacted bromophenol content). This material was successfully used without further purification.11 H NMR (CDCl3) δ 2.57 (t, 4H), 2.79 (t, 2H), 3.73 (t, 4H), 4.08 (t, 2H), 6.78 (d, 2H), 7.37 (d, 2H). MS (according to LC / MS): m / z 287.1 [M + 1]. The ease with which bromophenol can be removed has been shown in a 2 g sample by first dissolving the sample in toluene (8 g) and washing with 8 g of 15% aqueous NaOH; the product recovered by liquid chromatography. No trace of unreacted bromophenol was shown in the sample (1.97 g; 98.5% recovery).<chemistry num="16"><img id="000018" he="24" wi="84" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 6-fluoropyridin-3-ylboronic acid (4): Stirring Cooling (Dry Ice-Acetone Bath) Anhydrous [TBME] (620mL; in a 3L 3-neck round bottom flask with automatic stirrer, temperature probe with adapter, and nitrogen supply port) (by syringe) 2 M of BuLi (352 mL, 0.704 mol, 1.2 eq) was added. To this fast stirring and cooling (<-75 ° C) mixture was added 3 (102.2 g, 0.581 mol) anhydrous TBME (100 mL) solution over 13 minutes, during which the internal temperature rose to -62 ° C. The reaction mixture was further stirred for 45 minutes (temperature maintained at -62 ° C to -80 ° C), and then triisopropyl borate (total 180 g, 0.957 mol, 1.65 eq) was added rapidly and continuously in 4 portions. .. At the end of the addition, the internal temperature had risen to -33 ° C. After stirring in a cold water bath for an additional 45 minutes (internal temperature dropped from -33 ° C to -65 ° C), excluding the cold water bath, the agitated mixture naturally warmed to -22 ° C over 50 minutes. .. After heating to 6 ° C. over 15 minutes (by water bath), the stirring reaction mixture was placed in an ice water bath and then quenched with a solution of NaOH (160 g) in cold water (500 mL) under nitrogen. When the addition was complete, the internal temperature was 20 ° C. The mixture was stirred at room temperature for 1.5 hours. The aqueous layer was removed, neutralized to pH 7 with about 350 mL of concentrated HCl and then extracted with EtOAc (3 x 1 L). At this point, the pH was 8-9, so the aqueous layer was adjusted to pH 7 with about 15 mL of concentrated HCl and further (2 x 1 L) extracted with ethyl acetate. The combined EtOAc extracts are dried (Na)2SO4), Filtered and concentrated to a volume of about 150 mL. Divided addition of heptane (total volume 300 mL) with swirling of this concentrate resulted in product precipitation / crystallization. Filtration, washing of solids with heptane (100 mL, 300 mL, then another 300 mL), and air drying yielded off-white solids (68.6 g, yield 79-90%).*L C purity 96.4%, NMR showed an estimated 5.5% w / w heptane)), which was successfully used without further purification. LC / MS showed that this was a mixture of the following two entities, with the strength of the higher molecular weight being predominantly (*Note: The yield of the reaction is 79% assuming that boronic acid is the only constituent, and 90% if cyclic boronate is assumed to be the only constituent) :.<chemistry num="17"><img id="000019" he="52" wi="99" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>11 H NMR (CDCl3) δ 7.14 (dd, 1H), 8.27 (ddd, 1H), 8.39 (br s, 2H, 2 OH), 8.54 (fine d, 1 Η). MS (by LC / MS): m / z 143.0 [M + 1; boronic acid] and 370.0 [M + 1; cyclic boronate above].<chemistry num="18"><img id="000020" he="36" wi="134" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine (5): 2 (110.7g, 0.387mol), 4 (71.05g,) in a 2L 3-neck round bottom flask equipped with an automatic stirrer, thermometer and adapter, concentrator, and nitrogen supply port (top of concentrator) 0.477 mol (1.23 eq) and DME (700 mL) were charged. The resulting stirred solution was passed through a fast nitrogen stream through the stirred solution for 5 minutes and then Na.2CO3(121.06g, 1.142mol, 3 equivalents) H2O (250 mL) degassing solution as well as solid Pd (PPh)3)4Degassed by adding (19.8 g, 0.044 eq). Immediately after the last addition, nitrogen was purged into the headspace above the reaction mixture, then the mixture was stirred at 80-85 ° C (internal temperature) for 7 hours and then cooled to room temperature. Since there is no aqueous layer, decantation of the supernatant left inorganic salts (with adsorbed water). The reaction flask containing the inorganic salt was washed with 50% dichloromethane / ethyl acetate (2 x 250 mL) and the wash solution was added to the decanted supernatant. Dry this combined organic layer (Na2SO4), Filtered and evaporated to dryness to dark brown oil (148 g). To this oil, 150 g of 50% heptane / isopropyl alcohol (IPA) was added, swirled and cooled (by ice water bath), after which crystallization began. Further heptane (50 g) was added, and the obtained solid was filtered, washed, and air-dried to obtain 48 g of a light brown solid. After the filtrate is evaporated to dryness, the resulting mixture is swirled in 100 mL of 50% heptane / IPA, then heptane (about 100 mL) is added, stoppered, placed in a freezer and crystallized. It was. The resulting solid was filtered, washed with heptane and air-dried to give 61 g of gum-like solid. An oil (34 g) was obtained by evaporation of the obtained filtrate. Polar impurities contained in this (Ph3(P = O, etc.) was fairly low, so it was partitioned between 2N HCl (240 mL) and EtOAc (220 mL). Remove the bottom aqueous layer, then stir with EtOAc and K2CO3Neutralized to pH 7-8. The EtOAc layer was dried, filtered and evaporated to dryness (22 g). Some 48g, 61g and 22g were chromatographed on silica gel (1.1Kg) packed in DCM. DCM (400 mL), 50% DCM / EtOAc (5 L), then increasing doses of MeOH / Et3N (1.5% MeOH / 1% Et3Starting with N 5% MeOH / 3% Et3Elution with 50% DCM / EtOAc (8L) containing (ending in N) gave 77.68g of viscous oil (98.0% purity), which crystallized immediately by swirling in heptane (300mL). Was done. Filtration, washing with heptane and air drying yielded 75.55 g (98.7% AUC) of solid 5. More pure 5 (total weight 3.9g, 98.6-99.3% AUC), Ph3It was obtained from the first chromatographic fraction containing P = O by evaporative crystallization after washing as was done with the 34 g sample above. The total yield of 5 was 79.5 g (68%).11 H NMR (CDCl3) δ 2.59 (t, 4H), 2.84 (t, 2H), 3.75 (t, 4H), 4.16 (t, 2H), 6.97 (dd, 1H), 7.01 (d, 2H), 7.46 (d, 2H) , 7.92 (dd d, 1H), 8.37 (fine d, 1H). MS (according to LC / MS): m / z 303.2 [M + 1].<chemistry num="19"><img id="000021" he="33" wi="155" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) acetonitrile (6): A 3 L 3-necked round-bottom flask was equipped with an automatic stirrer, a thermometer and adapter, a dropping funnel, and a nitrogen supply port (upper part of the dropping funnel, positive pressure from a bubbler). The stopper was removed while passing a high-speed nitrogen stream through the bubbler, and KHMDS (415.8 g, 2.08 mol) and then anhydrous THF (1 L) were placed in a flask. Stir cooling (ice / methanol bath, internal temperature of the solution was -8 ° C) Immediately after dropping a solution of MeCN (70 g) in THF (110 mL) over 22 minutes into a KHMDS / THF solution, relatively rapidly ( For 4 minutes) 5 (79.06 g, 0.262 mol) of THF (400 mL) solution was added and after that period the internal temperature of the reaction mixture reached 10 ° C. With continuous cooling (1 hour), the internal temperature was set to -6 ° C and the reaction appeared to be complete according to TLC. After an additional 30 minutes (internal temperature -3 ° C), the reaction mixture was quenched with saturated brine (1 L) and diluted with EtOAc (500 mL). After removing the aqueous layer, the organic solution is dried (Na2SO4), Filter, evaporate to dryness (up to oil), dissolve completely in IPA (150 mL), dilute with heptane (300 mL), seed crystal (about 100 mg crude oil) IPA (about 150 mg) (Prepared by diluting with heptane (about 2.5 mL)) was added and left overnight. After stirring to decompose the crystalline solids, the solids are filtered, washed with 250 mL 2: 1 heptane / IPA, then washed multiple times with heptane and air dried to give the title 64.38 g (76% yield). Product 6 was obtained as a crystalline brown solid (LC purity 99.3%). An additional 5.88 g of low purity material was obtained from the filtrate.11 H NMR (CDCl3) δ 2.59 (t, 4H), 2.84 (t, 2H), 3.74 (t, 4H), 3.97 (s, 2H), 4.17 (t, 2H), 7.02 (d, 2H), 7.46 (d, 1H) , 7.51 (d, 2H), 7.87 (dd, 1H), 8.77 (fine d, 1H). MS (according to LC / MS): m / z 324.4 [M + 1].<chemistry num="20"><img id="000022" he="34" wi="159" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate (7): Put 6 (64.00g, 0.198mol) and MeOH (360g) in a 2L one-mouth round bottom flask, and then slowly and carefully H.2SO4(240 g) was added dropwise and the resulting homogeneous solution was refluxed (115 ° C oil bath) until the reaction was complete (25 hours (with 0.8% unreacted starting material)) 3.5% ArCH.2CO2Stirred with H. After cooling for a short time, DDL4(75 g) was added, the mixture was swirled and left for an additional 45 minutes (where the composition was 96.3% product, 0.8% unreacted starting material, and 2.5% ArCH.2CO2H). Then, the reaction mixture was added to DCM (2L) and K.2CO3(450g) H2It was added slowly to a fast stirring cooling (ice water bath) mixture with O (600 mL) solution. The resulting emulsion was left overnight. The clear portion of the organic solution was sucked up and the rest was repeatedly treated with water and DCM to combine the clear organic portion with the original portion that was sucked up. Dry the combined organic parts (Na2SO4), Filtered and concentrated to a volume of about 1.2 L, then 300 mL of 5% EtOAc (in heptane) followed by heptane (300 mL) was added to concentrate the mixture (rotational concentration with heating) and remove the DCM again. .. At this point, 15 mL of EtOAc was added and the high temperature mixture was swirled until crystallization began, continued swirling until crystallization was nearly complete, then left to cool to room temperature for complete crystallization. The solid was then filtered and washed with 300 mL of 5% EtOAc (in heptane) and heptane (100 mL), then completely air dried to give 57.74 g (82% yield) of 7 as a pale yellow solid. (98.9% AUC). An additional 3.94 g of clear product (97.9% AUC) was obtained from the filtrate (total yield 87%).11 H NMR (CDCl3) δ 2.60 (t, 4H), 2.84 (t, 2H), 3.74 (overlapping t and s, 6H), 3.89 (s, 2H), 4.17 (t, 2H), 7.01 (d, 2H), 7.34 ( d, 1H), 7.49 (d, 2H), 7.80 (dd, 1H), 8.74 (fine d, 1H). MS (by LC / MS): m / z 357.4 [M + 1].<chemistry num="21"><img id="000023" he="39" wi="160" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide (KX2-391 free base) 7 (61.4 g, 0.172 mol), benzylamine (55.6 g, 0.519 mol, 3 equivalents), and anhydrous anisole (300 g) were placed in a 1 L volume one-mouth round bottom flask, and then the reaction was essential under reflux. The mixture was stirred until it was completely finished (23 hours, oil bath temperature of 165 ° C; internal temperature was 147 ° C), and then allowed to cool to almost room temperature. Dilution of a portion (1 mL) of the reaction mixture with toluene (1 mL) resulted in complete crystallization of that portion. The seed crystals were then added to the reaction mixture and left to stand until the entire reaction mixture was crystallized into a single mass. Toluene (150 mL) was added and the mixture was swirled to decompose the solids. Heptane / toluene (1: 1, 100 mL) was added to further decompose the solid mixture. Finally, heptane (50 mL, then 25 mL) was added to further decompose the mixture, allowing for an additional 30 minutes and then filtering the solids. Filtration of solids, washing with 2: 1 toluene / heptane (300 mL), 1: 2 toluene / heptane (300 mL), then heptane (2 x 300 mL), then drying (air drying, then high vacuum) to 60.16 g The title product (yield 81%) was obtained as a white solid (> 98.9% AUC). An additional 2.5 g of low purity (97.4%) material was obtained from the mother liquor.11 H NMR (CDCl3) δ 2.60 (t, 4H), 2.83 (t, 2H), 3.74 (t, 4H), 3.82 (s, 2H), 4.18 (t, 2H), 4.49 (d, 2H), 7.01 (d, 2H) , 7.2-7.35 (m, 6H), 7.49 (d, 2H), 7.64 (br t, 1H), 7.81 (dd, 1H), 8.69 (fine d, 1H). MS (according to LC / MS): m / z 432.5 [M + 1].<chemistry num="22"><img id="000024" he="34" wi="161" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Preparation of 4- (2- (4- (6- (2- (benzylamino) -2-oxoethyl) pyridinium-3-yl) phenoxy) ethyl) -morpholine-4-ium chloride (KX2-391, diHCl salt) 170 mL of 2.5 M HCl (in ethanol) was added to an anhydrous EtOH (600 mL) stirred suspension of KX2-391 (free base, 60.00 g) and 25 mL of EtOH was added to flush the sides of the flask. .. The resulting homogeneous solution was stirred at room temperature (20 minutes) and then almost evaporated to dryness (until foaming). After tracking with EtOH (2 x 150 mL), the residue was dissolved again in EtOH (150 mL), then heptane was added slowly until the mixture appeared saturated (it took 33 mL to become cloudy). did. After being left overnight, two layers were formed. After further addition of heptane (250 mL), crystallization was still not induced, so the reaction mixture was concentrated to a volume of about 200 mL, at which point the mixture was homogeneous. This high concentration uniform solution was added dropwise to (automatic) EtOAc (2 L) stirring at very high speed. After completion of the dropping, 25 mL of the original flask and 25 mL of the dropping funnel of EtOH rinse was added to the fast stirring mixture. High speed stirring was continued for an additional hour, then the mixture was filtered and the solid (partially gum) was washed with EtOAc (300 mL) and then heptane. As soon as heptane washing was started, the solids became more gumy. The glass Buchner funnel and its contents were covered (paper towel / rubber band) and immediately placed in a vacuum furnace. After vacuuming overnight at about 45 ° C, evacuate under nitrogen, immediately place the Büchner funnel containing the product (foam solid) in a ziplock bag, then bottle under nitrogen (glove bag). The foamy solid was decomposed into powder (spatula). On the evening of the second day under high vacuum (about 45 ° C), an additional weight loss of only 1.3 g was achieved. On the evening of the third day of high vacuum (about 45 ° C), an essentially constant weight was obtained, with a weight loss of only 0.2 g. The final weight of the material is 68.05 g (97% yield), 0.29 equivalent (4.8% w / w) of EtOAc, 0.035 equivalent (0.3% w / w) of EtOH, and 0.03 equivalent (0. It contained 6% w / w) heptane. The purity was 99.6%.1H NMR (DMSO-d6) δ 3.1-3.3 (m, 2H), 3.45-3.65 (m, 4H), 3.8-4.0 (m, 4H), 4.11 (s, 2H), 4.32 (d, 2H), 4.57 (t, 2H), 7.19 (d, 2H), 7.2-7.4 (m, 5H), 7.88 (d, 2H), 7.93 (d, 1H), 8.68 (dd, 1H), 8.99 (br t, 1H), 9.10 (fine d, fine d, 1H), 11.8 (br s, 1H). MS (by LC / MS): m / z 432.5 [free base M + 1]. Elemental analysis (C26H29N3O3. 2HCl . 0.035EtOH . 0.29 EtOAc . 0.03 heptane . 0.8H2O's): Calculated (%): C, 60.03; H, 6.54; N, 7.65; Cl, 12.91 b. Measured value (%): C, 59.85 / 59.97; H, 6.54 / 6.47; N, 7.67 / 7.67; Cl, 13.10 / 13.24 Calculated FW: 534.63 (probably 0.8H generated when handling this highly hygroscopic powder2O is11 NMR is H2Not considered because it showed no evidence of O). The ethyl chloride level in this material was measured and found to be 98 ppm. The sample was also analyzed and found to contain 5,800 ppm heptane. Analysis of another part of this sample gave the following results: 99.6% AUC, 1640 ppm ethanol, 41,480 ppm ethyl acetate, 5600 ppm heptane, no anisole detected, and 120 ppm ethyl chloride. In addition, the procedure for recrystallization of the salt was developed using the above-mentioned dried salt. This procedure will work quite well as it would for the high purity crude salt (containing residual EtOH) obtained by concentrating the HCl salt-forming reaction mixture. The salt (575 mg) was dissolved in anhydrous EtOH (1.157 g) in double mass and then heated under nitrogen. To this high temperature solution (stirring), 1.6 g of 25% EtOH (in EtOAc) was added, and then EtOAc (0.25 mL) was added, resulting in a turbid state. When this turbid high temperature solution was allowed to cool to room temperature, crystallization occurred during this period. After crystallization is complete (2 hours), the crystalline solid is filtered, washed with anhydrous EtOAc (approximately 40 mL) and dried in vacuo to give 424 mg of KX2-391 dihydrochloride as a free-flowing solid. (Fine beads, 99.8% AUC), which contained only 0.05 equivalents (0.45% w / w) of EtOH and 0.015 equivalents (0.26% w / w) of EtOAc. Somewhat better recovery (586 mg to 460 mg) was obtained with isopropanol / EtOAc, but solvent capture levels were high [0.085 eq (1.0% w / w) of isopropanol and 0.023 eq (0.4% w / w). ) EtOAc]. Example 3: Large-scale synthesis of KX2-391 di HCl Reagents and solvents were used as received from commercial sources. The progress of the reaction is HPLC, GC / MS, or1Monitored by 1 H NMR. Thin layer chromatography (TLC) was performed using an Analtech silica gel plate and visualized by UV light (254 nm). High performance liquid chromatography (HPLC) was performed on an Agilent 1100 series instrument. Proton and carbon nuclear magnetic resonance spectra were acquired using Bruker AV 300 at 300 MHz for protons and 75 MHz for carbon. Solvent peaks were used as reference peaks in the proton and carbon spectra. Preparation of 4- (2- (4-Bromophenoxy) Ethyl) Morpholine (2) 4- (3-Chloropropyl) morpholine (2.44 kg, 0.54 mol), 4-bromophenol (2.27 kg, 0.54 mol, 1.0 eq), in a 50 L jacketed reactor equipped with a reflux concentrator and a temperature probe. Powdered potassium carbonate (6.331 kg, 1.88 mol, 3.50 eq) and DMF (12.2 L) were charged and stirred. The reaction mixture was then heated to 60-65 ° C and stirred overnight. After 17.5 hours, the reaction mixture was cooled to 20-25 ° C. The reaction mixture was charged into different reactors equipped with a bottom valve for preparation. DI water (48.7L) was charged into this reactor while maintaining the temperature at 20-30 ° C. Phase separation has occurred. The aqueous layer was extracted with MTBE (3 × 24.4L). DI water (18.3 L) followed by 6M sodium hydroxide (18.2 L) was added to the combined organic layers. Stirring the mixture for 2-5 minutes resulted in phase separation. The organic phase was washed with water (24.4 L) and brine (24.4 L), dried over magnesium sulphate, filtered, concentrated and 3370 g of yellow oil (89% crude yield, 99.4% AUC by HPLC). Preparation of 6-fluoropyridin-3-ylboronic acid (4) A reflux concentrator and a temperature probe were attached to a 72 L reactor. The reactor was charged with 5-bromo-2-fluoropyridine (1.17 L, 0.568 mol), toluene (18.2 L), and triisopropyl borate (3.13 L, 0.68 mol, 1.2 eq) and stirred. Tetrahydrofuran (4.4 L) was added to the reactor and the reaction mixture was cooled to -35 to -50 ° C. N-Butyllithium (2.5 M hexane solution, 5.44 L, 0.68 mol, 1.2 eq) was carefully added to the reactor while maintaining the temperature at -35 to -45 ° C. After 5 hours, the reaction was considered complete and the reaction mixture was warmed to -15 to -20 ° C. In this reaction solution of the reactor, 2M HCl (11.80 L) was added and the temperature was maintained at -15 ° C to 0 ° C. Stirring of the reaction mixture at 18-23 ° C (16 hours) resulted in phase separation. The organic phase was then extracted with 6M sodium hydroxide (6.0L). The acidic non-basic aqueous phase was mixed in the reactor and 6M HCl (2.5L) was added until pH 7.5 was reached. Sodium chloride (6.0 kg) was then added to the aqueous phase. The aqueous phase was then extracted with THF (3 × 20 L). The combined organic moieties were dried over magnesium sulphate and concentrated to give 1300 g of brown solids (81% crude yield). Preparation of 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine (5) 6-Fluoropyridine-3-ylboric acid (2.84 kg, 1.24 eq), 4- (2- (4-bromophenoxy)) in a 72 L reactor equipped with a reflux concentrator, sparging tube, bubbler, and temperature probe. Ethyl) morpholine (4.27 kg, 1.0 eq) and DME (27 L) were charged. Stirring was started and then sodium carbonate (4.74 kg, 3.0 eq) was charged into the reaction mixture as a solution of DI water (17.1 L). Argon was foamed in the reaction mixture for 50 minutes. Tetrakis (triphenylphosphine) palladium (750 g, 0.04 eq) was added to the reaction mixture as a slurry in DME (1.0 L) under an argon atmosphere. The reaction mixture was heated to 75-85 ° C and stirred overnight (17 hours). The reaction mixture was cooled to 18-22 ° C. DI water (26.681 kg) and MTBE (26.681 L) were charged into the reactor and stirred for 5 minutes. Phase separation occurred and the aqueous phase was extracted with MTBE (2 × 26.7 L). 2M of combined organic parts Extracted with HCl (1 × 15.0L, 3 × 21.8L). Then, the aqueous phase was returned to the reactor and charged, and ethyl acetate was added (26.7 L). The pH was adjusted to 6.2 with 6M sodium hydroxide (26.7L) and the temperature was maintained at 15-25 ° C. Phase separation occurred and the aqueous phase was extracted with ethyl acetate (2 x 26.7 L). The combined organic moieties were dried over magnesium sulphate and concentrated to give 4555 g of residue (101% crude yield, 67.1% AUC by HPLC). Purification of 4- (2- (4- (6-fluoropyridin-3-yl) phenoxy) ethyl) morpholine (5) The crude product (575 g) was purified by silica gel chromatography (methanol / ethyl acetate / heptane (30% ethyl acetate / heptane, 50% ethyl acetate / heptane, 75% ethyl acetate / heptane, 100% ethyl acetate, and 5%). Eluted with methanol / ethyl acetate). TLC (10% methanol / dichloromethane, RfElution of the pure fraction by = 0.3) gave 420 g of light brown solid (73% recovery,> 99.9% AUC by HPLC). Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) acetonitrile (6) Preparation of 1M NaHMDS (2.0L, 5.0 eq) THF was placed in a 5L flask and cooled to -20 to -15 ° C. The flask was charged with fluoride (119.7 g, 1.0 eq) in THF (500 mL) over 20 minutes, keeping the temperature below -10 ° C. THF (170 mL) containing acetonitrile (82.5 mL, 4.0 eq) was added to the flask over 20 minutes and the temperature was maintained below -10 ° C. The reaction mixture was then stirred for 1 hour. Brine (1.5 L, 12.6 vol) was added to the reaction at a rate that kept the temperature below 10 ° C. The solution was then warmed to room temperature for layer separation. The mixture was filtered over Celite and washed with THF (1 x 200 mL, 1 x 100 mL). The aqueous phase was extracted with toluene (750 mL). The combined organic moieties were dried over magnesium sulfate, filtered, washed with toluene (2 x 25 OmL) and concentrated to dryness. Toluene (1 L) was added and the solution was concentrated to dryness again to give 169.8 g of oil. MTBE (1190 mL, 7 vol) was added to the oil at 50 ° C and stirred for 15 minutes. Heptane (850 mL, 5 volumes) was added at 50 ° C for 10 minutes. The mixture was then cooled to room temperature over 1.5 hours and stirred for 2 hours. The slurry was filtered, washed with 1: 4 MBTE / heptane (2 x 100 mL) and dried in the oven at 45 ° C overnight to give 102.3 g of off-white solid (80% yield, 80% yield, 98.8% AUC by HPLC). Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate (7) Nitrile 6 (101 g) and methanol (1.01 L, 10 volumes) were placed in a 3 L flask equipped with a stirring rod and a thermocouple. Concentrate H in this solution2SO4(175 mL, 10.0 eq) was added dropwise over 15 minutes and the temperature was maintained below 60 ° C. Subsequently, 30% fuming sulfuric acid (124 mL) was added dropwise to this solution, and the temperature was maintained below 60 ° C. The solution was then reflux heated in a heating mantle and stirred overnight. When the reaction was considered complete, it was cooled to 20 ° C. Saturated sodium bicarbonate (10.7 L) and dichloromethane (1.1 L) were placed in a second flask (22 L) and cooled to 15 ° C. The reaction mixture was added to the sodium bicarbonate / dichloromethane mixture while maintaining the temperature below 20 ° C. Phase separation occurred when the quench solution was stirred for 15 minutes. The aqueous phase was extracted with dichloromethane (1 x 550 mL, 1 x 300 mL). The combined organic moieties were dried over magnesium sulphate and concentrated to dryness to give 105 g of orange solid (94% crude yield, 97.7% AUC by HPLC). Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide (KX2-391) Ester 7 (103 g), anisole (513 mL, 5 volumes), and benzylamine (94 mL, 3.0 eq) were placed in a 3 L flask equipped with a thermocouple and an overhead stirrer. The reaction mixture was then heated to 142 ° C. and stirred for 2 days. The reaction mixture was cooled to 45-50 ° C and stirred for 2 hours. To this mixture, n-heptane (1.5 L) was added dropwise over 1 hour. The solution was cooled to room temperature over 3 hours and then stirred overnight. The resulting slurry was filtered and washed with 4: 1 anisole / n-heptane (200 mL) and n-heptane (3 x 100 mL). After drying overnight in the oven, the product obtained was 112.1 g brown solid (90% yield, 99.6% AUC by HPLC). The use of a single isomer of heptane was essential to adequately quantify the residual solvent. KX2-3911See Figure 5 for 1 H NMR. Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide dihydrochloride (KX2-391 2HC1) EtOH (1.0 L) was placed in a 2 L flask, and acetyl chloride (62.5 mL, 3.0 eq) was slowly added to the flask, and the mixture was stirred for 40 minutes. The resulting solution was added to KX2-391 (100 g) over 30 minutes and the temperature was maintained at 30 ° C. The solution was concentrated to a mass of 270 g. The concentrated solution was added to ethyl acetate (2 L) over 20 minutes with high speed stirring. The mixture was stirred overnight and then filtered under nitrogen to give two different solid products, a brown solid (73.5 g) and a darker solid (42.2 g). These solids were dry blended to give a total yield of 99%. HPLC analysis showed 99.0% purity (AUC). Analysis showed that the presence of ethanol was 2530 ppm, ethyl acetate was 48,110 ppm, ethyl chloride was 170 ppm, and heptane and anisole were not detected. The palladium content was assayed three times and measured to be less than 29 ppm, 2 ppm, and 1 ppm. Crystallization test of KX2-391 . 2HCl The experiments shown in Table 1 were performed to investigate various crystallization and precipitation conditions of KX2-391 . 2HCl.<tables num="1-1"><img id="000025" he="115" wi="167" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="1-2"><img id="000026" he="203" wi="167" file="JP2017193587A_D0001.tif" img-format="tif" img-content="drawing" /></tables> Precipitation was carried out by back-adding KX2-391 / 2HC1 to a large volume of fast stirring ethyl acetate in a concentrated ethanol solution. This precipitation procedure was performed in a demonstration batch that resulted in the formation of two different types of solids. The two different types of solids were physically separated and filtered separately. The light brown solid (lot 02BP111E, 74 g, 99.1% AUC by HPLC) was first filtered, and then the dark brown solid (lot 02BP111F, 43 g, 99.1% AUC by HPLC) was filtered. After drying in a vacuum furnace, each of the two solid samples was held for analysis before blending. The data of interest are differential scanning calorimetry (DSC, Figures 1 and 2) and powder X-ray diffraction (XRPD, Figures 3 and 4). The HPLC data for the two samples were similar, but the DSC and XRPD were different. Both HPLC preparations had a purity greater than 99.0% (by area%), with one endothermic event at approximately 198 ° C for the lot 02BP111E sample, compared to 117 ° C and 189 for the lot 02BP111F sample. Two endothermic events were shown at ° C. The XRPD data of the two samples were also different, with the lot 02BP111E sample appearing to be crystalline, while the lot 02BP111F sample appeared to be amorphous. HPLC data, XRPD data and DSC data confirm that the two samples are in different forms of the same material. These two lots of KX2-391 / 2HCl (lot 02BP111E and 02BP111F) were dry-blended to obtain a new lot of KX2-391 / 2HCl (lot 02BP111G). KX2-391 . 2HCl (lot 02BP111G) contained 170 ppm ethyl chloride. Example 4: Preparation of 2- (5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate (KX2-391 . MSA) Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) acetonitrile (6) Sodium bis (trimethyldisilyl) amide (1.0 M, 23.2 L in THF) was charged into the round bottom reactor 1 and the solution was cooled to -10 ° C over 52 minutes. Compound 5 (1400 g, 1 weight) and THF (7.0 L, anhydrous, 5 volumes)) were charged into a glass carboy under nitrogen. The batch was stirred under nitrogen with an air driven stirrer. This batch was not completely soluble and was a cloudy solution. A solution of compound 5 was added to reactor 1 from a 5 L dropping funnel over 41 minutes. A solution of acetonitrile (965 mL, anhydrous, 0.69 vol) in THF (2.0 L, anhydrous, 1.43 vol) was prepared and added to Reactor 1 from the same dropping funnel at -10 ° C over 48 minutes (trace amount of yellow). Solids were present on the wall of the reactor). After aging at -10 ° C for 45 minutes, batch sampling was performed for analysis and compound 5 was 0.03% by conversion (standard is 1.5% by conversion). After 1 hour and 24 minutes of sampling, brine (17.6 L, 12.6 vol) was added to Reactor 1 over 52 minutes to give an inadequately stirred batch (similar to an emulsion). A diatomaceous earth pad was prepared on a 24-inch polypropylene funnel (1026 g Celite 545 3.3 L water slurry, filtrate exhaust). The batch was filtered from the pad under suction, the reactor was rinsed with THF (1.75 L, 1.25 volume) and the rinse was transferred to the cake. The cake was rinsed with a second portion of THF (1.75 L, 1.25 volume) and the total filtration time was 1 hour 17 minutes. The filtrate was transferred to reactor 2, phase separated and retained overnight (batch was retained under nitrogen in the reactor). Discard the organic phase (approximately 34.5L) and replace the aqueous phase with toluene (8.1L, 5. 8 volumes), stirred for 16 minutes and left for 12 minutes. After separation, it is also possible to omit the toluene extraction and simply add toluene directly to the organic phase. The aqueous phase (approximately 19 L) was removed, the organic phases were combined and dried over 55 minutes in Magnesium Sulfate (1400 g, 1 weight, anhydrous) in Reactor 2. Batches were filtered through a 24-inch polypropylene funnel fitted with an in-line filter into a glass carboy. The batch was covered with argon and stored in a cold room (2-8 ° C) (concentration pending). The next day, the batch was concentrated to a residue, rinsed with toluene (11.8 L, 8.4 vol) and concentrated (water bath 50 ± 5 ° C). At the time of toluene addition, the batch was an orange slurry and remained intact after concentration. The total concentration time was 5 hours and 3 minutes. Reactor 3 was charged with MTBE (13.9 L, 9.9 volume, ACS) and then heated to 45 ± 5 ° C. MTBE was discarded and approximately 2 L of MTBE was used for slurry batch from valve into reactor 3. The remaining MTBE was added to reactor 3 and the batch was maintained at 45 ± 5 ° C, then the batch was aged in this temperature range for 33 minutes. N-heptane (10 L, 7.1 volume, 99%) was then added to reactor 3 over 39 minutes to maintain the batch at 45 ± 5 ° C. The heating was turned off and the batch was cooled to 25 ± 5 ° C for 4 hours and 5 minutes and aged in this temperature range for 27 hours and 4 minutes. The batch was then filtered under aspiration with a 24-inch polypropylene funnel (PTFE cloth), covered under dry nitrogen and aspirated. The total filtration time was 20 minutes. The orange batch (net wet weight 1322 g) was dried in a vacuum furnace set at 45 ± 5 ° C for 48 hours and 3 minutes until a constant weight was reached. The batch was transferred to two 80 oz amber glass bottles (sealed with Teflon®) and covered with argon (6 of 1217 g, 81% of theoretical value). Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) methyl acetate (7) Compound 6 (900 g, 2.78 mol) and methanol (9.0 L, 10 volumes, anhydrous) were charged into a 22 L reactor. Sulfuric acid (1115 mL, fuming) was added to this suspension over 2 hours 11 minutes to give a dark solution. The maximum temperature was 65.5 ° C (target 65 ° C). Sulfuric acid (1565 mL, 1.74 vol, concentrated) was added to the batch over 1 hour 49 minutes, then the batch was heated to visible reflux (74 ° C) for 18 minutes. The batch was maintained at that temperature for 16 hours 57 minutes. If this mild visible reflux was found to be absent, the batch was reflux-heated again at 79-80 ° C for 2 hours and 15 minutes. Keep the batch at that temperature (80 ± 5 ° C) for 10 hours 57 minutes, then turn off the heating and after 26 hours 4 minutes add an additional load of methanol (0.75 L, 0.8 volume, anhydrous) and lose. The volume of solvent was replenished. It was estimated that 2.5-3.3 L of solvent was lost by evaporation. HPLC analysis 42 hours and 31 minutes after reflux showed that the level of compound 6 was 0.6% by conversion (standard 1.0%). Methylene chloride (4.8 L, 5.3 volume) and sodium hydrogen carbonate solution (48 L, 53.3 volume, saturated) were charged into each of reactors 1 and 2. The sodium hydrogen carbonate solution was stored overnight at 2-8 ° C and removed the next morning. Half batches of 22 L reactors were divided and added to each reactor over 47 and 44 minutes, respectively (batch temperatures were 12-13 ° C and 14-15 ° C, respectively). Quenching was performed by the generation of carbon dioxide (vigorous agitation with vortex). The batch of each reactor was then transferred to a 200 L reaction vessel, the batch was stirred for 16 minutes and then left for 25 minutes to separate the organic phase. The aqueous phase was continuously extracted twice with methylene chloride (5 L, 5.6 volumes, and 2.7 L, 3 volumes); Each extraction was carried out over 15 minutes with stirring and left for 6 and 9 minutes, respectively. The combined organic phases were transferred to Reactor 3 and dried over magnesium sulfate (900 g, 1 weight, anhydrous) for 35 minutes. The batch was then filtered under suction with a 24-inch polypropylene funnel fitted with sharkskin cloth and an in-line filter (10 micron, Pall P / N 12077). The filtrate was concentrated on a rotary evaporator at 40 ± 5 ° C (water bath temperature) for a total of 2 hours and 18 minutes. After 54 minutes, the batch solidified and spheroids were formed. This was decomposed and concentration was continued. The batch (a mixture of fine solids and brittle masses) was then further ground and returned to the valve for continued concentration. The batch was transferred to an 80 ounce amber jar with a Teflon® covered lid and covered with argon to give compound 7 (871 g, 88% of theory). Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide (KX2-391) A 22 L reactor was charged with compound 7 (650 g, 1.82 mol), anisole (3.25 L, 5 volumes, anhydrous) and benzylamine (600 mL, 0.92 volumes, 3 eq). The batch (approximately 18 ° C) was heated to 142 ± 5 ° C for 1 hour and 44 minutes and dissolution occurred at 30 ° C. The batch was maintained at 142 ± 5 ° C for 69 hours and 30 minutes, at which point HPLC analysis showed that compound 7 was 0.9% by conversion (standard: 1.7% by conversion). The batch was cooled to 45-50 ° C for 5 hours and 12 minutes (increased nitrogen flow when the batch reached approximately 72 ° C to aid cooling). In that temperature range, the batch was poorly agitated and when mixed, the batch temperature rose to 52 ° C. This was> 50 ° C for 15 minutes. When first <50 ° C, the batch is aged for 2 hours 2 minutes, then n-heptane (9.75 L, 15 volumes, 99%) is added to the batch over 1 hour 56 minutes to adjust the batch temperature. Maintained at 45-50 ° C. The heating was then stopped and the batch was cooled to 25 ° C for 10 hours and 32 minutes and then to approximately 20 ° C for 20 minutes. The total time to maintain the batch at 25 ° C was 4 hours 50 minutes (2 hours 47 minutes at approximately 20 ° C). The batch was filtered under suction with a 24-inch polypropylene filter funnel (attached with PTFE cloth), the reactor was rinsed with anisole / n-heptane (1.3 L, 4: 1) and the rinse was transferred to the cake. The cake was then washed twice (1.3 L, 0.65 L) continuously with n-heptane. The total filtration time was 39 minutes. The batch (KX2, 391 with a net wet weight of 1004 g) was transferred to three glass trays, placed in a vacuum furnace set at 50 ° C. and dried for 96 hours and 26 minutes until a constant weight was reached. Preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) Pyridine-2-yl) -N-benzylacetamide mesylate (KX2-391 . MSA) KX2-391 (520 g, 1.21 mol) was transferred to Reactor 1 with acetone (41.6 vol, 80 vol, ACS) for ease of transfer. When the batch was heated to 50 ± 5 ° C for 33 minutes, dissolution occurred at 30 ° C. Batch inline filter (Pall P / N It was entirely transferred into the second reactor by a transport pump equipped with 12077, 10 microns) and reheated from 46 ° C to 50 ± 5 ° C. Methanesulfonic acid (121.4 g, 1.05 eq, 99% ultrapure) was added to the pale yellow batch over 12 minutes and then heating was interrupted. After 14 minutes, a white solid was observed, which increased in number, and after 59 minutes, a white suspension was obtained. The batch became 25 ± 5 ° C after 7 hours and 51 minutes, and was further aged for 19 hours and 21 minutes (10 hours and 30 minutes at 27 ° C). The batch was filtered under suction with a 24-inch polypropylene filter (PTFE cloth), the reactor was rinsed with acetone (2.0 L, clarification, ACS) and the rinse was transferred to the cake. The cake was covered with a stainless steel cover and sucked under a stream of dry nitrogen. The total filtration time was 21 minutes. The batch (net wet weight 764 g) was transferred to three glass drying trays and dried in a vacuum furnace at 25 ± 5 ° C for 21 hours 54 minutes (565 g, 89% of theory). The sample was removed for analysis and the batch was maintained in vacuo at 25 ± 5 ° C. The batch was then transferred to two 80 ounce amber glass bottles (polypropylene encapsulation coated with Teflon®), covered with argon and stored at -10 to -20 ° C. Other embodiments The present invention also provides: (1) (1) Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to obtain 4- (2- (4-bromophenoxy) ethyl) morpholine; (2) Coupling 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-Il) Phenoxy) Ethyl) Morpholine Obtaining Step; (3) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate The process of converting to methyl; and (5) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) by reacting methyl acetate with benzylamine Step to obtain pyridine-2-yl) -N-benzylacetamide A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (2) (1) Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to obtain 4- (2- (4-bromophenoxy) ethyl) morpholine; (2) Coupling 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-Il) Phenoxy) Ethyl) Morpholine Obtaining Step; (3) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile; (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate Step to convert to methyl; (5) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) by reacting methyl acetate with benzylamine Step to obtain pyridine-2-yl) -N-benzylacetamide; and (6) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide is contacted with methanesulfonic acid to make 2- (5- (4- (2-morpholino) phenyl) Step to obtain ethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate, which comprises. (3) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide is contacted with methanesulfonic acid to make 2- (5- (4- (2-morpholino) phenyl) Step to obtain ethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate, which comprises. (4) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine-2-yl) 2-(5- (4- (2-morpholinoethoxy) phenyl) by reacting methyl acetate with benzylamine Step to obtain pyridine-2-yl) -N-benzylacetamide A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (5) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate Step to convert to methyl 4. The method of item 4 for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (6) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile 5. The method of item 5 for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (7) Coupling 4- (2- (4-bromophenoxy) ethyl) morpholine with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-Il) Phenoxy) Ethyl) Morpholine 6. The method of item 6 for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (8) Step of reacting 4- (2-chloroethyl) morpholine with 4-bromophenol to obtain 4- (2- (4-bromophenoxy) ethyl) morpholine A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (9) 4- (2- (4- (4- (6-Fluoropyridine) ethyl) morpholine was coupled with 6-fluoropyridin-3-yl-3-boronic acid to 4- (2- (4- (6-fluoropyridine)- 3-yl) phenoxy) ethyl) morpholine to obtain the A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (10) 4- (2- (4- (6-Fluoropyridine-3-yl) phenoxy) ethyl) morpholine is reacted with acetonitrile to make 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridine- 2-Il) Step to obtain acetonitrile A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (11) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetonitrile with 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) acetate Step to convert to methyl A method for the preparation of 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, which comprises. (12) 2- (5- (4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide is contacted with methanesulfonic acid to make 2- (5- (4- (2-morpholino) phenyl) 2-(5-(4- (2-morpholinoethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide, including the step of obtaining ethoxy) phenyl) pyridin-2-yl) -N-benzylacetamide mesylate. The method according to any one of items 4 to 11 for the preparation of mesylate. (13) A composition containing KX2-391 mesylate. (14) The composition according to item 13, wherein the KX2-391 mesylate has a purity higher than 98.0% when measured by HPLC. (15) The composition according to item 13, wherein the KX2-391 mesylate has a purity of 99.0%. (16) The composition according to item 13, wherein the KX2-391 mesylate has a purity of 99.5%. (17) The composition according to item 13, wherein the KX2-391 mesylate has a purity of 99.6%. (18) The composition according to item 13, wherein the KX2-391 mesylate has a purity of 99.7%. (19) The composition according to item 13, wherein the composition comprises less than 2% impurities selected from ethyl chloride, ethanol, ethyl acetate, heptane, anisole, palladium, and combinations thereof. (20) The composition according to any one of items 13 to 19, further comprising a pharmaceutically acceptable carrier or excipient. (21) Use of the composition according to any one of items 13 to 20 in the manufacture of a pharmaceutical for regulating one or more components of a protein kinase signaling cascade. (22) The use according to item 21, wherein the medicament inhibits a kinase selected from Src family protein kinases, focal adhesion kinases, and tyrosine kinases. (23) The use according to item 22, wherein the tyrosine kinase is a Src family protein kinase. (24) The use according to any one of items 21 to 23, wherein the drug is orally administered. (25) The use according to any one of items 21 to 23, wherein the drug is locally administered. (26) Diseases or disorders in which the components of the kinase cascade are selected from hyperproliferative disorders, cancer, precancer, osteoporosis, cardiovascular disorders, immune system dysfunction, type II diabetes, obesity, deafness, and graft rejection. The use according to item 21, which is the cause of the onset of symptoms. Although the present invention has been described with its detailed description, the above description is intended to be exemplary and does not limit the scope of the invention as defined in the appended claims. Other aspects, advantages and variations are included in the appended claims. Those skilled in the art will appreciate that various modifications can be made herein in form and in detail without departing from the scope of the invention within the scope of the appended claims.

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Document	Office	Kind	Date
60930758	United States of America	–	2007-05-17
93075807	United States of America	P	2007-05-17
93075807	United States of America	P	2007-05-17
12005792	United States of America	–	2007-12-28
579207	United States of America	A	2007-12-28
579207	United States of America	A	2007-12-28
12005792	–	–	–
60930758	–	–	–
US20070005792	–	–	–
US20070930758P	–	–	–

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US1462426A	United States of America	A	1923-07-17
US4539629A	United States of America	A	1985-09-03
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CA2673974A1	Canada	A1	2008-07-10
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CN101616915A	China	A	2009-12-30
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CN101687798A	China	A	2010-03-31
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AU2014202221B2	Australia	B2	2016-06-23
MY158648A	Malaysia	A	2016-10-31
JP6022657B2	Japan	B2	2016-11-09
JP2016222733A	Japan	A	2016-12-28
CA2673974C	Canada	C	2017-01-17
US9556120B2	United States of America	B2	2017-01-31
US2017101378A1	United States of America	A1	2017-04-13
JP2017193587AThis record	Japan	A	2017-10-26
EP2155681B1	European Patent Office (EPO)	B1	2018-05-02
JP6329871B2	Japan	B2	2018-05-23
DK2155681T3	Denmark	T3	2018-08-06
ES2682108T3	Spain	T3	2018-09-18
US10323001B2	United States of America	B2	2019-06-18
US2019256468A1	United States of America	A1	2019-08-22
US2021198202A1	United States of America	A1	2021-07-01
US2024343691A1	United States of America	A1	2024-10-17

3 legal events, as the office reported them to INPADOC

Over the term

Point at a mark for the event

Events

Legal events, newest first
	Event	Code
2018-11-29	Decision of refusalJAPANESE INTERMEDIATE CODE: A02A02	A02
2018-05-08	Notification of reasons for refusalJAPANESE INTERMEDIATE CODE: A131A131	A131
2017-08-04	Written request for application examinationJAPANESE INTERMEDIATE CODE: A621A621	A621