Hydroxylamine-based epidermal growth factor receptor inhibitors for treatment of brain metastatic cancer

Abstract

According to the purposes of the present disclosure, as embodied and broadly described herein, the disclosure relates, in one aspect, to scaffold molecules that inhibit epidermal growth factor receptor (EGFR), methods of making the same, pharmaceutical compositions comprising the same, and methods of treating cancers involving aberrant EGFR activity.

Description

[0001] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with government support under Grant No. R21 GM144753 awarded by the National Institutes of Health. The government has certain rights in this invention. (37 CFR 401.14f(4)).

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims the benefit of and priority to co-pending U.S. Provisional Patent Application Serial No. 63 / 375,622 filed on September 14, 2022 and U.S. Provisional Patent Application Serial No. 63 / 507,876 filed on June 13, 2023, the entire contents of which are incorporated herein by reference. Background Art

[0005] Lung cancer remains the leading cause of cancer-related deaths worldwide, with an estimated 2.2 million new cases and 1.79 million deaths each year. 1 Epidermal growth factor receptor (EGFR, erbB1, HER-1) is an attractive molecular target for lung cancer drug development. Activating mutations, such as the point mutation L858R on exon 21 and the in-frame exon 19 deletion delE746_A750, are detected in 10% to 30% of patients with non-small cell lung cancer (NSCLC), rendering them sensitive to reversible first-generation EGFR-targeted tyrosine kinase inhibitors (TKIs) such as gefitinib (1). Figure 1-2 ) 2-5 Unfortunately, in NSCLC, up to 40% of patients develop brain metastases (BM), a number that is likely to increase as treatment options continue to improve life expectancy for patients with advanced disease. 6 Multiple studies have shown that most marketed EGFR TKIs have low CNS penetration due to active efflux by transporters such as P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), which are highly enriched at the blood-brain barrier (BBB) ​​and bind to tight junctions, ultimately excluding up to 98% of the drug from the CNS ( Figure 1 ) 7-10 Therefore, future treatment options for NSCLC will benefit from improved CNS management, enabling treatment of both localized disease and subsequent brain metastases. Summary of the invention

[0006] In accordance with the purposes of the present disclosure, as embodied and broadly described herein, the present disclosure relates in one aspect to scaffold molecules that inhibit epidermal growth factor receptor (EGFR), methods of making the same, pharmaceutical compositions comprising the same, and methods of treating cancers involving aberrant EGFR activity.

[0007] Other systems, methods, features and advantages of the present disclosure will be or become apparent to those skilled in the art by inspection of the following drawings and detailed description. All of these additional systems, methods, features and advantages are included in this specification, are within the scope of the present disclosure, and are protected by the appended claims. In addition, all optional and preferred features and modifications of the described embodiments can be used in all aspects of the disclosure taught herein. In addition, the various features of the dependent claims, as well as all optional and preferred features and modifications of the described embodiments, are combinable and interchangeable. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Many aspects of the present disclosure may be better understood with reference to the following drawings. The components in the drawings are not necessarily drawn to scale, with emphasis placed on clearly illustrating the principles of the present disclosure. In addition, in the drawings, the same reference numerals represent corresponding parts in multiple views.

[0009] Figure 1 Shown are the chemical structures of selected approved EGFR tyrosine kinase inhibitors (TKIs) and their efflux states against efflux transporters.

[0010] Figure 2 New bioisostere modifications are shown.

[0011] Figure 3 An alternative synthetic route to the hydroxylamine precursor (17) is shown.

[0012] Figure 4 The synthesis of hydroxylamine-based EGFR inhibitors is shown (6).

[0013] Figure 5 and Figure 6 Exemplary procedures for preparing compounds described herein are shown.

[0014] Figure 7 Shown are the hydroxylamine-based EGFR inhibitors described herein.

[0015] Figure 8 The synthesis of hydroxylamine-based EGFR inhibitors is shown (15)

[0016] Table 1 shows the in vitro ADMET properties of gefitinib and hydroxylamine-based EGFR inhibitors. Unless otherwise stated, values ​​represent the average of n=2 independent replicates. Ames results were negative at concentrations from 15 to 50 μM, after which bacterial cytotoxicity was observed. Aq.Sol. indicates water solubility; LMCl int Indicates the intrinsic clearance in liver microsomes; HEPCl int represents the intrinsic clearance rate of hepatocytes; t 1 / 2P represents half-life; app =MDCK stands for apparent permeability; MDCK stands for Madine-Darby-canine-kidney; MDR1 stands for multidrug resistance 1 (or P-glycoprotein). Abbreviations: H stands for human; R stands for rat; nd stands for not determined.

[0017] Fig. 9 The in vitro antiproliferative activity of gefitinib and hydroxylamine-based EGFR inhibitors is shown. Hydroxylamine-based EGFR inhibitors (15) showed potent antiproliferative activity in patient-derived non-small cell lung cancer cell lines HCC827 (EGFR mutant = exon 19del), NCI-H3255 (EGFR mutant = L858R), and osimertinib-resistant engineered cell lines (Ba / F3-L858R / C797S; Ba / F3-del E746_A750 / C97S), but showed no activity in NCI-H1975 (EGFR mutant = L85858R / T790M) and in A431 cell line (overexpressing EGFR wt ) shows the minimum activity (72 h administration period). For all antiproliferative assays, points represent mean values ​​and error bars represent standard deviations; n=3 independent replicates; IC 50 Values ​​(nM) were calculated from dose-response curves and reported as mean ± standard error (SEM).

[0018] Table 2 shows the in vitro ADMET properties of compound 15. The values ​​represent the average of n≥2 independent replicates. The in vitro micronucleus negative test results were effective at 31 μM with rat liver S9 and 8 μM without rat liver S9, and then the cytotoxicity was observed. Aq.Sol. indicates water solubility; f u,plasma % indicates the percentage of unbound in plasma; f u,brain % indicates the percentage of unbound in the brain; HEPCl int Indicates the intrinsic clearance rate of hepatocytes. Abbreviations: H for human; R for rat; C for cynomolgus monkey; D for dog.

[0019] Fig.10 The results of the non-mutant kinase or lipid kinase screening of compound 15 by KINOMEScan are shown at a screening concentration of 1 M. On the kinase phylogenetic tree drawn using DiscoverX TREEspot, the size of the circle is proportional to the strength of the binding affinity.

[0020] Fig.11 The subsequent binding (Kd) and biochemical inhibition (IC) of compound 15 to the kinase are shown. 50), these kinases showed <10% activity in KINOMEScan. Results are the average of n=2 independent replicates.

[0021] Fig.12 The in vitro subsequent antiproliferative activity of compound 15 in breast cancer cell lines with different HER2 status is shown. Compound 15 showed moderate antiproliferative activity in patient-derived HER2-positive breast cancer cell lines AU565, SK-BR-3, BT474 and ZR-75-30. For all antiproliferative assays, points represent mean values ​​and error bars represent standard deviations; n=3 independent replicates; IC 50 Values ​​(nM) were calculated from dose-response curves and reported as mean ± standard error (SEM).

[0022] Fig.13 Shown are total plasma versus time profiles (0 to 24 hours) following single dose administration of Compound 15 to SD rats at 2 mg / kg IV and 20 mg / kg PO and to BALB / c nude mice at 3 mg / kg IV and 30 mg / kg PO. AUC 0-inf (nM·h) represents the area under the concentration-time curve from 0 to ∞; t 1 / 2 (h) represents the mean elimination half-life obtained by intravenous infusion (IV) or oral gavage (PO); F (%) represents bioavailability (%); T max (h) represents the time to reach peak plasma concentration; C max (nM) represents the peak plasma concentration; CL (mL·min -1 kg), clearance obtained by intravenous infusion. For pharmacokinetic curves, points represent mean values ​​and error bars represent standard deviations; n=3 animals per route (n=6 in total). Values ​​are expressed as mean ± standard error.

[0023] Fig.14 The central nervous system pharmacokinetic profile of hydroxylamine-based EGFR inhibitors is shown (15). Total plasma and brain concentration versus time curves (0 to 24 hours) 15 hours after a single dose of 20 mg / kg in SD rats and 40 mg / kg in CD1 mice by oral gavage (PO) showed excellent brain penetration. Kp brain (AUC inf ) refers to the partition coefficient of brain to plasma. Kp,uu brain (AUC inf ) refers to the partition coefficient of unbound brain to unbound plasma. For pharmacokinetic curves, points represent mean values ​​and error bars represent standard deviations; n=3 animals per time point (n=21 total).

[0024] Fig.15Bioluminescent images of the vehicle control group and the compound 15 (10 mg / kg, orally twice daily) administration group are shown, indicating the changes in tumor volume within 21 days of treatment.

[0025] Fig.16 The changes in intracranial bioluminescence of mice over the course of the study as a function of the time of administration of compound 15 and the changes in the average body weight of mice are shown. For the bioluminescence graph, the points represent the mean and the error bars represent the SEM; P=0.0059; n=10 mice per group. PDX, patient-derived xenograft. The p value was obtained by an unpaired two-tailed t-test comparing the mean values ​​of the solvent control group and the compound 15 (10 mg / kg, oral twice daily) study group after 21 days of treatment. *P<0.05, **P<0.01. For the average body weight graph, the points represent the mean and the error bars represent the SEM; P=0.2275; n=10 mice per group. The p value was obtained by an unpaired two-tailed t-test comparing the mean values ​​of the solvent control group and the compound 15 (10 mg / kg, oral twice daily) study group after 21 days of treatment. Ns indicates not significant.

[0026] Figure 17-18 An exemplary synthetic route to hydroxylamine-based nitrogen heterocycles is shown.

[0027] Other advantages of the present invention will be described in part in the following description, and in part will be apparent from the description, or can be learned through the practice of the present invention. The advantages of the present invention will be realized and obtained through the elements and combinations specifically pointed out in the appended claims. It should be understood that the above general description and the following specific embodiments are only exemplary and explanatory and do not limit the invention claimed. DETAILED DESCRIPTION

[0028] For those skilled in the art to which the disclosed compositions and methods belong, numerous improvements and other embodiments disclosed herein can be imagined with the benefit of the teachings presented in the foregoing description and the associated drawings. Therefore, it should be understood that the disclosed content is not limited to the specific embodiments disclosed, and modifications and other embodiments are intended to be included within the scope of the appended claims. Those skilled in the art will recognize many variations and adaptations of the aspects described herein. These variations and adaptations are intended to be included in the teachings of the present disclosure and are covered by the claims herein.

[0029] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0030] Those skilled in the art will appreciate after reading this disclosure that each individual embodiment described and illustrated herein has discrete components and features that can be readily separated or combined with the features of any one of the other several embodiments without departing from the scope or spirit of the present disclosure.

[0031] Any recited method may be performed in the order of events recited or in any other order that is logically possible. That is, unless expressly stated otherwise, any method or aspect described herein is in no way intended to require that its steps be performed in a particular order. Therefore, if a method claim does not specifically state in the claim or specification that the steps are to be limited to a particular order, no order should be inferred in any respect. This applies to any possible non-express basis for interpretation, including logical issues related to the arrangement of steps or operational flows, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0032] All publications mentioned herein are incorporated herein by reference to disclose and describe methods and / or materials citing these publications. The publications discussed herein are only used to disclose before the date of filing of the present application. Anything herein shall not be construed as admitting that the present invention is not entitled to be earlier than the relevant publications by virtue of prior invention. In addition, the publication date provided herein may be different from the actual publication date, which may require independent confirmation.

[0033] Although aspects of the present disclosure may be described and claimed in specific legal categories, such as the systems legal category, this is for convenience only and those skilled in the art will understand that each aspect of the present disclosure may be described and claimed in any legal category.

[0034] It should also be understood that the terms used herein are only used to describe specific aspects and are not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the disclosed compositions and methods belong. It should be further understood that terms (such as those defined in commonly used dictionaries) should be interpreted as having the same meaning as their meanings in the specification and the context of the relevant technology, and unless clearly defined herein, should not be interpreted in an idealized or overly formal sense.

[0035] Before describing the various aspects of the present disclosure, the following definitions are provided, which should be used unless otherwise indicated. Additional terms may be defined elsewhere in this disclosure.

[0036] definition

[0037] As used herein, "comprising" should be interpreted as specifying the presence of the features, integers, steps or components mentioned, but does not exclude the presence or addition of one or more features, integers or components or groups thereof. In addition, each of the terms "by", "comprising", "comprises", "consisting of", "including", "includes", "included", "involving", "involves", "involved", and "for example" are used for open, non-restrictive purposes and can be used interchangeably. In addition, the term "comprising" is intended to include examples and aspects covered by the terms "consisting essentially of" and "consisting of". Similarly, the term "consisting essentially of" is intended to include instances of "consisting of".

[0038] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "an excipient" includes but is not limited to a mixture or combination of two or more such excipients, and the like.

[0039] It should be noted that ratios, concentrations, amounts, and other numerical data may be expressed in a range format herein. It should be further understood that the endpoints of each range, whether associated or independent relative to another endpoint, are of substantial significance. It should also be understood that many values ​​are disclosed herein, and in addition to the value itself, each value is also disclosed herein as "approximately" for the particular value. For example, if the value "10" is disclosed, "approximately 10" is also disclosed. The range may be expressed here as from "approximately" a particular value and / or to "approximately" another particular value. Similarly, when a value is expressed as an approximation, by using the antecedent "approximately", it is understood that the particular value forms another aspect. For example, if the value "approximately 10" is disclosed, "10" is also disclosed.

[0040] When expressing a range, another aspect includes from a specific value and / or to another specific value. For example, if the range includes one or two limitations, the disclosure also includes a range that does not include one or both of these limitations, for example, the phrase "x to y" includes a range from "x" to "y" and greater than "x" and less than "y". The range can also be expressed as an upper limit, such as "about x, y, z or less", which should be interpreted as including a specific range of "about x", "about y" and "about z", as well as a range of "less than x", "less than y" and "less than z". Similarly, the phrase "about x, y, z or greater" should be interpreted as including a specific range of "about x", "about y" and "about z", as well as a range of "greater than x", "greater than y" and "greater than z". In addition, the phrase "about 'x' to 'y'", where 'x' and 'y' are numerical values, includes "about 'x' to about 'y'".

[0041] It should be understood that this range format is used for convenience and brevity and, therefore, should be interpreted in a flexible manner to include not only the values ​​explicitly listed as range limits, but also all individual values ​​or sub-ranges contained within the range, as if each value and sub-range were explicitly listed. For illustration, a numerical range of "about 0.1% to 5%" should be interpreted to include not only the explicitly cited values ​​of about 0.1% to about 5%, but also individual values ​​(e.g., about 1%, about 2%, about 3%, and about 4%) and sub-ranges (e.g., about 0.5% to about 1.1%; about 5% to about 2.4%; about 0.5% to about 3.2% and about 0.5% to about 4.4%, as well as other possible sub-ranges) within the indicated range. Thus, for example, if the amount of a component is about 1%, 2%, 3%, 4%, or 5%, any value of which can be the lower and upper endpoints of the range, any range between 1% and 5% (e.g., 1% to 3%, 2% to 4%, etc.) can be envisioned.

[0042] As used herein, the terms "approximately," "equal to or about," and "substantially" mean that the amount or value in question may be an exact value or a value that provides an equivalent result or effect as described in the claims or taught herein. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and features are not and need not be exact, but may be approximate and / or larger or smaller as needed, reflecting tolerances, conversion factors, rounding, measurement errors, etc., as well as other factors known to those skilled in the art, so as to obtain an equivalent result or effect. In some cases, it is not possible to reasonably determine a value that provides an equivalent result or effect. In such cases, as used herein, it is generally understood that, unless otherwise stated or inferred, "about" and "at or about" refer to a variation of ±10% of the nominal value shown. In general, whether or not explicitly stated, an amount, size, formulation, parameter, or other quantity or feature is considered to be "about," "approximate," or "at or about." It is understood that where "about," "approximate," or "at or about" is used before a quantitative value, the parameter also includes the specific quantitative value itself, unless specifically stated otherwise.

[0043] As used herein, “IC 50 ” refers to the concentration of a substance (such as a compound or drug) required to inhibit a biological process or component of a process by 50%. For example, IC 50 It refers to the half-maximal (50%) inhibitory concentration (IC) of a substance as determined in an appropriate assay.

[0044] A chemical substance residue as used in the specification and the summarized claims refers to the product portion of a chemical substance in a particular reaction scheme or subsequent preparation or chemical product, whether or not that portion is actually obtained from the chemical substance. Thus, an ethylene glycol residue in a polyester refers to one or more -OCH2CH2O- units in the polyester, whether or not ethylene glycol was used to prepare the polyester. Similarly, a sebacic acid residue in a polyester refers to one or more -CO(CH2)8CO- moieties in the polyester, whether or not that residue was obtained by reacting sebacic acid or an ester thereof to obtain the polyester.

[0045] As used herein, the term "substituted" is intended to include all allowed substituents of organic compounds. In a broad sense, allowed substituents include acyclic rings and cyclic, branched and unbranched, carbocyclic and heterocyclic and aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, for example, those described below. Allowed substituents can be one or more, and can be the same or different for appropriate organic compounds. For the purposes of this disclosure, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any allowed substituents of organic compounds that meet heteroatom valences as described herein. This disclosure is not intended to be limited in any way by the allowed substituents of organic compounds. In addition, the term "substituted" or "substituted" includes implicit proviso that such substitution complies with the allowed valences of the substituted atoms and substituents, and such substitution produces stable compounds, such as compounds that do not spontaneously undergo rearrangement, cyclization, elimination, etc. It is also expected that, in some aspects, unless explicitly stated to the contrary, a single substituent can be further optionally substituted (i.e., further substituted or unsubstituted).

[0046] The position of a substituent may be defined relative to the position of other substituents in the aromatic ring. For example, as shown below, with respect to an "R" group, a second substituent may be "ortho," "para," or "meta" to the R group, meaning that the second substituent is bonded to the ortho, para, or meta position of the carbon marker, as shown below. Combinations of ortho, para, and meta substituents for a given group or substituent are also contemplated and should be considered disclosed.

[0047]

[0048] In defining various terms, “A 1 ”, “A 2 ”, “A 3 ” and “A 4 " are used as general symbols herein to indicate various specific substituents. These symbols may be any substituents, not limited to those disclosed herein, and when they are defined as certain substituents in one instance, they may be defined as some other substituents in another instance.

[0049] As used herein, the term "aliphatic" or "aliphatic group" refers to a hydrocarbon moiety that may be straight chain (i.e., unbranched), branched, or cyclic (including fused, bridged, and spiro-fused polycyclics), and may be fully saturated, or may contain one or more unsaturated units, but is not aromatic. Unless otherwise indicated, an aliphatic group contains 1-20 carbon atoms. Aliphatic groups include, but are not limited to, straight or branched alkyl, alkenyl, and alkynyl, and their hybrids, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0050] The term "alkyl" as used herein is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, etc. The alkyl group can be cyclic or non-cyclic. The alkyl group can be branched or unbranched. The alkyl group can also be substituted or unsubstituted. For example, the alkyl group can be substituted by one or more groups, including but not limited to alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxyl, nitro, silyl, sulfoxy or mercaptan, as described herein. "Low alkyl" refers to an alkyl group containing 1 to 6 (e.g., 1 to 4) carbon atoms. The term alkyl may also be C1 alkyl, C1-C2 alkyl, C1-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C7 alkyl, C1-C8 alkyl, C1-C9 alkyl, C1-C10 alkyl, etc., up to and including C1-C24 alkyl.

[0051] Throughout the specification, "alkyl" is generally used to refer to unsubstituted alkyl and substituted alkyl; however, by identifying specific substituents on the alkyl, substituted alkyl is also specifically mentioned herein. For example, the term "halogenated alkyl" or "haloalkyl" specifically refers to an alkyl substituted by one or more halides, such as fluorine, chlorine, bromine or iodine. Alternatively, the term "monohaloalkyl" specifically refers to an alkyl substituted by a single halide, such as fluorine, chlorine, bromine or iodine. The term "polyhaloalkyl" specifically refers to an alkyl independently substituted by two or more halides, i.e., each halide substituent does not need to be the same halide as another halide substituent, nor does it require multiple instances of a halo substituent to be on the same carbon. The term "alkoxyalkyl" specifically refers to an alkyl substituted by one or more alkoxy groups, as described below. The term "aminoalkyl" specifically refers to an alkyl substituted by one or more amino groups. The term "hydroxyalkyl" specifically refers to an alkyl substituted by one or more hydroxyl groups. When "alkyl" is used in one case and a specific term such as "hydroxyalkyl" is used in another case, it does not mean that the term "alkyl" does not also refer to a specific term such as "hydroxyalkyl".

[0052] This approach also applies to other groups described herein. That is, while the term "cycloalkyl" refers to unsubstituted and substituted cycloalkyl moieties, substituted moieties may also be specifically identified herein; for example, a specific substituted cycloalkyl may be referred to as, for example, "alkylcycloalkyl." Similarly, a substituted alkoxy may be specifically referred to as, for example, "haloalkoxy," and a specific substituted alkenyl may be, for example, "enol," etc. Likewise, the use of a general term such as "cycloalkyl" and a specific term such as "alkylcycloalkyl" does not mean that the general term does not include the specific term.

[0053] The term "cycloalkyl" as used herein is a non-aromatic carbonyl ring consisting of at least three carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, etc. The term "heterocycloalkyl" is a cycloalkyl as defined above, and is included in the meaning of the term "cycloalkyl", wherein at least one carbon atom of the ring is replaced by a heteroatom, such as but not limited to nitrogen, oxygen, sulfur or phosphorus. Cycloalkyl and heterocycloalkyl can be substituted or unsubstituted. Cycloalkyl and heterocycloalkyl can be substituted by one or more groups, including but not limited to alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxyl, nitro, silyl, sulfoxy or thiol as described herein.

[0054] The term "alkanediyl" as used herein refers to a divalent saturated aliphatic group having one or two saturated carbon atoms as points of attachment, having a straight or branched chain, cyclo, cyclic or acyclic structure, no carbon-carbon double or triple bonds, no atoms other than carbon and hydrogen. The groups -CH2- (methylene), -CH2CH2-, -CH2C(CH3)2CH2- and -CH2CH2CH2 are non-limiting examples of alkanediyl groups.

[0055] As used herein, the terms "alkoxy" and "alkoxyl" may refer to an alkyl or cycloalkyl group attached through an ether bond; that is, "alkoxy" may be defined as -OA 1 , where A 1 is an alkyl or cycloalkyl group as described above. "Alkoxy" also includes the alkoxy polymers just described; that is, the alkoxy group can be a polyether, such as -OA 1 -OA 2 or—OA 1 —(OA 2 ) a —OA 3 , where "a" is an integer from 1 to 200, A 1 , A 2 and A 3 is an alkyl group and / or a cycloalkyl group.

[0056] As used herein, the term "alkenyl" is a hydrocarbon group of 2 to 24 carbon atoms having a structural formula with at least one carbon-carbon double bond. 1 A 2 )C=C(A 3 A 4) is intended to include both E and Z isomers. This may be inferred in the present structural formulas where an asymmetric olefin is present, or may be explicitly indicated by the bond symbol C=C. The alkenyl group may be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfoxy, or thiol, as described herein.

[0057] The term "cycloalkenyl" as used herein is a non-aromatic carbon-based ring consisting of at least three carbon atoms and containing at least one carbon-carbon double bond, i.e., C=C. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, norbornenyl, etc. The term "heterocycloalkenyl" is a cycloalkenyl group as defined above and is included within the meaning of the term "cycloalkenyl" in which at least one carbon atom of the ring is replaced by a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. Cycloalkenyl and heterocycloalkenyl groups may be substituted or unsubstituted. Cycloalkenyl and heterocycloalkenyl groups may be substituted by one or more groups, including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfoxy, or thiol as described herein.

[0058] As used herein, the term "alkynyl" is a hydrocarbon group of 2 to 24 carbon atoms having a structural formula containing at least one carbon-carbon triple bond. Alkynyl groups can be unsubstituted or substituted with one or more groups, including but not limited to alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfoxy or thiol, as described herein.

[0059] The term "cycloalkynyl" as used herein is a non-aromatic carbonyl ring consisting of at least seven carbon atoms and containing at least one carbon-carbon triple bond. Examples of cycloalkynyl include, but are not limited to, cyclooctynyl, cyclononynyl, etc. The term "heterocycloalkynyl" is a cycloalkenyl as defined above, included in the meaning of the term "cycloalkynyl", wherein at least one carbon atom of the ring is replaced by a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus. Cycloalkynyl and heterocycloalkynyl may be substituted or unsubstituted. Cycloalkynyl and heterocycloalkynyl may be substituted by one or more groups, including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfoxy or thiol as described herein.

[0060] The term "aromatic group" as used herein refers to a ring structure having a delocalized π-electron annular cloud above and below the plane of the molecule, wherein the π cloud contains (4n+2) π electrons. Aromaticity is further discussed in Morrison and Boyd (Organic Chemistry, (5th Ed., 1987), Chapter 13, entitled "Aromaticity," pages 477-497), which is incorporated herein by reference. The term "aromatic group" includes aryl and heteroaryl groups.

[0061] The term "aryl" as used herein is a group containing any carbon-based aromatic group, including but not limited to benzene, naphthalene, phenyl, biphenyl, anthracene, etc. The aryl group can be substituted or unsubstituted. The aryl group can be substituted by one or more groups, including but not limited to alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, ─NH2, carboxylic acid, ester, ether, halide, hydroxyl, ketone, azide, nitro, silyl, sulfoxy or thiol as described herein. The term "biaryl" is a specific type of aryl group, which is included in the definition of "aryl". In addition, the aryl group can be a single ring structure or can include multiple ring structures, which are fused ring structures or connected by one or more bridging groups (such as carbon-carbon bonds). For example, a biaryl group refers to two aryl groups, which are bound together by a fused ring structure, such as naphthalene, or connected by one or more carbon-carbon bonds, such as biphenyl. Fused aryl groups are also considered, including but not limited to indene and naphthyl.

[0062] The term "aldehyde" used herein is represented by the formula -C(O)H. In the present specification, "C(O)" is an abbreviation of a carbonyl group, that is, C=O.

[0063] As used herein, the term "amine" or "amino group" is represented by the formula: NA 1 A 2 Indicates that A 1 and A 2 It can be independently hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein. A specific example of an amino group is -NH2.

[0064] The term "alkylamino" as used herein is represented by the formula -NH(-alkyl) and -N(-alkyl)2, wherein alkyl is as described herein. Representative examples include, but are not limited to, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, (sec-butyl)amino, (tert-butyl)amino, pentylamino, isopentylamino, (tert-pentyl)amino, hexylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di(sec-butyl), di(tert-butyl)amino, dipentylamino, diisobutylamino, di(tert-pentyl)amino, dihexylamino, dimethylamino, dihexylamino, N-ethyl-N-methylamino, N-methyl-N-propylamino, N-ethyl-N-propylamino, and the like.

[0065] As used herein, the term "carboxylic acid" is represented by the formula— C(O)OH.

[0066] The term "ester" as used herein is represented by the formula -OC(O)A 1 or—C(O)OA 1 Indicates that A 1 It may be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein.

[0067] As used herein, the term "ether" is a 1 OA 2 Indicates that A 1 and A 2 It may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein.

[0068] As used herein, the terms "halo," "halogen," or "halide" are used interchangeably to refer to F, Cl, Br, or I.

[0069] As used herein, the terms "halide-like," "pseudohalogen," or "pseudohalo" are used interchangeably to refer to functional groups that behave substantially similarly to halides. For example, such functional groups include cyano, thiocyanate, azido, trifluoromethyl, trifluoromethoxy, perfluoroalkyl, and perfluoroalkoxy.

[0070] The term "heteroalkyl" as used herein refers to an alkyl group containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, and S, wherein nitrogen, phosphorus, and sulfur atoms are optionally oxidized, and nitrogen heteroatoms are optionally quaternized. The heteroalkyl group may be substituted by an alkyl group as defined above.

[0071] The term "heteroaryl" as used herein refers to an aromatic group incorporating at least one heteroatom within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus, wherein N-oxides, sulfur oxides, and dioxides are permitted heteroatom substitutions. Heteroaryl groups may be substituted or unsubstituted. Heteroaryl groups may be substituted by one or more groups, including but not limited to alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxyl, nitro, silyl, sulfoxy, or thiol as described herein. Heteroaryl groups may be monocyclic or fused ring systems. Heteroaryl includes, but is not limited to, furanyl, imidazolyl, pyrimidinyl, tetrazolyl, thienyl, pyridinyl, pyrrolyl, N-methylpyrrolyl, quinolyl, isoquinolyl, pyrazolyl, triazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridazinyl, pyrazinyl, benzofuranyl, benzodioxazole, benzothienyl, indolyl, indazolyl, benzimidazolyl, imidazopyridinyl, pyrazolopyridinyl, and pyrazolopyrimidinyl. Other non-limiting examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, pyrazolyl, imidazolyl, benzo[d]oxazolyl, benzo[d]thiazolyl, quinolinyl, quinazolinyl, indazolyl, imidazo[1,2-b]pyridazinyl, imidazo[1,2-a]pyrazinyl, benzo[c][1,2,5]thiadiazolyl, benzo[c][1,2,5]oxadiazolyl, and pyrido[2,3-b]pyrazinyl.

[0072] As used herein, the terms "heterocycle" or "heterocyclyl" are used interchangeably and refer to monocyclic and polycyclic aromatic or non-aromatic ring systems in which at least one ring member is not carbon. Thus, the term includes, but is not limited to, "heterocycloalkyl", "heteroaryl", "bicyclic heterocycle", and "polycyclic heterocycle". Heterocycles include pyridine, pyrimidine, furan, thiophene, pyrrole, isoxazole, isothiazole, pyrazole, oxazole, thiazole, imidazole, oxazole including 1,2,3-oxadiazole, 1,2,5-oxadiazole and 1,3,4-oxadiazole, thiadiazole (including 1,2,3-thiadiazole, 1,2,5-thiadiazole and 1,3,4-thiadiazole), triazole (including 1,2,4-triazole and 1,3,5-triazole), tetrazole (including 1,2,3,4-tetrazole and 1,2,4,5-tetrazole), pyridazine, pyrazine, triazine (including 1,2,4-triazine and 1,3,5-triazine), tetrazine (including 1,2,4,5-tetrazine), pyrrolidine, piperidine, piperazine, morpholine, azetidine, tetrahydropyran, tetrahydrofuran, dioxane, and the like. The term heterocyclic radical can also be C2 heterocyclic radical, C2-C3 heterocyclic radical, C2-C4 heterocyclic radical, C2-C5 heterocyclic radical, C2-C6 heterocyclic radical, C2-C7 heterocyclic radical, C2-C8 heterocyclic radical, C2-C9 heterocyclic radical, C2-C10 heterocyclic radical, C2-C11 heterocyclic radical, etc., up to and including C2-C18 heterocyclic radical. For example, C2 heterocyclic radical includes a group with two carbon atoms and at least one heteroatom, including but not limited to aziridine radical, diazetidinyl, dihydrodiazetyl, oxirane, thiirane, etc. Or, for example, C5 heterocyclic radical includes a group with five carbon atoms and at least one heteroatom, including but not limited to piperidinyl, tetrahydropyranyl, tetrahydrothiopyran, diazepanyl, pyridinyl, etc. It is understood that the heterocyclyl group may be bonded via a heteroatom in the ring (where chemically possible) or via a carbon comprising the heterocyclyl ring.

[0073] As used herein, the term "bicyclic heterocycle" or "bicyclic heterocyclyl" refers to a ring system in which at least one ring member is not carbon. Bicyclic heterocyclyls include ring systems in which an aromatic ring is fused to another aromatic ring, or in which an aromatic ring is fused to a non-aromatic ring. Bicyclic heterocyclyls include ring systems in which a benzene ring is fused to a 5- or 6-membered ring containing 1, 2, or 3 ring heteroatoms or ring systems in which a pyridine ring is fused to a 5- or 6-membered ring containing 1, 2, or 3 ring heteroatoms. Bicyclic heterocyclic groups include, but are not limited to, indolyl, indazolyl, pyrazolo[1,5-a]pyridinyl, benzofuranyl, quinolinyl, quinoxalinyl, 1,3-benzodioxinyl, 2,3-dihydro-1,4-benzodioxinyl, 3,4-dihydro-2H-chromenyl, 1H-pyrazolo[4,3-c]pyridin-3-yl; 1H-pyrrolo[3,2-b]pyridin-3-yl; and 1H-pyrazolo[3,2-b]pyridin-3-yl.

[0074] The term "heterocycloalkyl" as used herein refers to aliphatic, partially unsaturated or fully saturated 3 to 14 ring systems, including monocycles and bicyclic and tricyclic systems of 3 to 8 atoms. The heterocycloalkyl ring system includes one to four heteroatoms independently selected from oxygen, nitrogen and sulfur, wherein nitrogen and sulfur heteroatoms are optionally oxidized, and nitrogen heteroatoms are optionally substituted. Representative heterocycloalkyl includes but is not limited to pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl and tetrahydrofuranyl.

[0075] As used herein, the term "hydroxyl" or "hydroxy" is represented by the formula -OH.

[0076] The term "ketone" as used herein is represented by the formula A 1 C(O)A 2 Indicates that A 1 and A 2 It may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein.

[0077] As used herein, the term "azide" or "azido" is represented by the formula -N3.

[0078] The term "nitro" as used herein is represented by the formula -NO2.

[0079] As used herein, the term "nitrile" or "cyano" is represented by the formula -CN.

[0080] The term "silyl" as used herein is represented by the formula: SiA 1 A 2 A 3 Indicates that A 1 , A2 and A 3 It may independently be hydrogen or an alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein.

[0081] The term "sulfoxy" as used herein is represented by the formula: S(O)A 1 ,—S(O)2A 1 、—OS(O)2A 1 or—OS(O)2OA 1 Indicates that A 1 It can be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein. In this specification, "S(O)" is an abbreviation for S=O. The term "sulfoxy" is used herein to refer to a sulfoxyl group of the formula: S(O)2A 1 A represents a sulfoxyl group, wherein A 1 It can be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein. The term "sulfone" as used herein is represented by the formula A 1 S(O)2A 2 Indicates that A 1 and A 2 can be independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl as described herein. The term "sulfoxide" as used herein is a sulfoxide of formula A 1 S(O)A 2 Indicates that A 1 and A 2 It may independently be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein.

[0082] As used herein, the term "thiol" is represented by the formula -SH.

[0083] “R 1 ”, “R 2 ”, “R 3 ”……“R n ", wherein n is an integer, as used herein, may independently have one or more of the groups listed above. For example, if R 1 If the first group is a straight chain alkyl group, one of the hydrogen atoms of the alkyl group may be optionally substituted by a hydroxyl group, an alkoxy group, an alkyl group, a halide, etc. Depending on the selected group, the first group may be incorporated into the second group, or the first group may be attached (i.e., connected) to the second group. For example, using the phrase "an alkyl group containing an amino group", the amino group may be incorporated into the backbone of the alkyl group. Alternatively, the amino group may be connected to the backbone of the alkyl group. The nature of the selected group will determine whether the first group is embedded in or connected to the second group.

[0084] As described herein, the compounds of the present invention may contain "optionally substituted" moieties. In general, the term "substituted", whether or not preceded by the term "optionally", means that one or more hydrogens of the designated moiety are replaced by suitable substituents. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when multiple positions in any given structure may be substituted by multiple substituents selected from a particular group, the substituents may be the same or different at each position. The combinations of substituents contemplated by the present invention are preferably those that result in the formation of stable or chemically feasible compounds. It is also contemplated that, in certain aspects, unless expressly to the contrary, a single substituent may be further optionally substituted (i.e., further substituted or unsubstituted).

[0085] As used herein, the term "stable" refers to a compound that is not substantially altered when placed under conditions that allow for its production, detection, and, in some aspects, its recovery, purification, and use for one or more of the purposes disclosed herein.

[0086] Suitable monovalent substituents on the substitutable carbon atoms of the "optionally substituted" group are independently halogen; -(CH2) 0- 4R o ; -(CH2) 0-4 OR o ; -O(CH2) 0-4 R o ; -O-(CH2) 0-4 C(O)OR o ; -(CH2) 0-4 CH(OR o )2; -(CH2) 0-4 SR o ; -(CH2) 0-4 Ph, which can be o Substitution; -(CH2) 0-4 O(CH2) 0-1 Ph, which can be o Substitution; -CH=CHPh, which can be replaced by R o Substitution; -(CH2) 0-4 O(CH2) 0-1 -pyridine, which can be o Substitution; -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R o )2; -(CH2) 0-4 N(R o )C(O)R o ;-N(R o )C(S)R o ; -(CH2) 0-4 N(Ro )C(O)NR o 2;-N(R o )C(S)NR o 2;-(CH2) 0-4 N(R o )C(O)OR o ;-N(R o )N(R o )C(O)R o ;-N(R o )N(R o )C(O)NR o 2;-N(R o )N(R o )C(O)OR o ;-(CH2) 0-4 C(O)R o ;-C(S)R o ;-(CH2) 0-4 C(O)OR o ;-(CH2) 0-4 C(O)SR o ;-(CH2) 0-4 C(O)OSiR o 3;-(CH2) 0-4 OC(O)R o ;-OC(O)(CH2) 0-4 SR-;SC(S)SR o ;-(CH2) 0-4 SC(O)R o ;-(CH2) 0-4 C(O)NR o 2;-C(S)NR o 2;-C(S)SR o ;-(CH2) 0- 4OC(O)NR o 2;-C(O)N(OR o )R o ;-C(O)C(O)R o ;-C(O)CH2C(O)R o ;-C(NOR o )R o ;-(CH2) 0-4 SSR o ;-(CH2) 0-4 S(O)2R o ;-(CH2) 0-4 S(O)2OR o ;-(CH2) 0-4 OS(O)2R o; -S(O)2NR o 2; -(CH2) 0-4 S(O)R o ;-N(R o )S(O)2NR o 2;-N(R o )S(O)2R o ;-N(OR o )R o ; -C(NH)NR o 2;-P(O)2R o ;-P(O)R o 2;-OP(O)R o 2;-OP(O)(OR o )2;SiR o 3;-(C 1-4 Straight or branched alkylene)ON(R o )2 or -(C 1-4 Straight or branched alkylene)C(O)ON(R o )2, where each R o may be substituted as defined below and are independently hydrogen, C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, -CH2-(5-6 membered heteroaromatic ring) or a 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definitions, two independent occurrences of R o Together with their central atoms they form a 3-12 membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0087] R o A suitable monovalent substituent on (or two independent occurrences of R o The ring formed by the middle atom thereof) is independently halogen, -(CH2) 0-2 R · 、-(haloR · )、-(CH2) 0-2 OH, -(CH2) 0-2 OR · 、-(CH2) 0-2 CH(OR · )2. -O(haloR · )、-CN、-N3、-(CH2) 0-2 C(O)R · 、-(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR· 、-(CH2) 0-2 SR · 、-(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR · 、-(CH2) 0-2 NR · 2. -NO2, -SiR · 3. -OSiR · 3. -C(O)SR · 、-(C 1-4 Straight or branched alkylene)C(O)OR · or -SSR · , wherein each R is unsubstituted or preceded by "halogen" at a position substituted with only one or more halogens and is independently selected from C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. o Suitable divalent substituents on a saturated carbon atom of include =0 and =S.

[0088] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0, =S, =NNR * 2. =NNHC(O)R * 、=NNHC(O)OR * 、=NNHS(O)2R * , =NR * , =NOR * 、-O(C(R * 2)) 2-3 O-or-S(C(R * 2)) 2-3 S-, wherein R* is independently selected from hydrogen, C 1-6 aliphatic, or an unsubstituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents bonded to adjacent substitutable carbons of the "optionally substituted" group include: -O(CR * 2) 2-3 O-, where R * Each independent occurrence of is selected from hydrogen, C substituted by 1-6 Aliphatic, or unsubstituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0089] R* Suitable substituents on the aliphatic group include halogen, -R · 、-(haloR · ),-OH, -OR · 、-O(haloR · )、-CN、-C(O)OH、-C(O)OR · 、-NH2、-NHR · 、-NR · 2 or -NO2, wherein each R is unsubstituted or substituted only with one or more halogens at the position preceded by "halogen" and is independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0090] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include or Each of these is independently hydrogen, C which may be substituted as defined below 1-6 aliphatic, unsubstituted -OPh, or unsubstituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definitions, two independent occurrences of Together with their central atoms, they form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0091] Suitable substituents on the aliphatic group are independently halogen, -R · 、-(haloR · ), -OH, -OR · 、-O(haloR · )、-CN、-C(O)OH、-C(O)OR · 、-NH2、-NHR · 、-NR · 2 or -NO2, where each R · is unsubstituted or substituted only by one or more halogens at the positions preceding it and is independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0092] The term "leaving group" refers to an atom (or group of atoms) with electron-withdrawing ability that can be substituted as a stable species and takes away bonding electrons. Examples of suitable leaving groups include halides and sulfonates, including but not limited to trifluoromethanesulfonate, mesylate, p-toluenesulfonate and p-bromobenzenesulfonate.

[0093] The compounds described herein may contain one or more double bonds and may therefore give rise to cis / trans (E / Z) isomers and other conformational isomers. Unless otherwise specified, the present invention includes all such possible isomers, as well as mixtures of such isomers.

[0094] Unless otherwise indicated, chemical formulas that are represented by solid lines only and not by wedges or dashed lines are understood to encompass every possible isomer, such as each enantiomer and diastereomer, as well as mixtures of isomers, such as racemic mixtures or meso mixtures. The compounds described herein may contain one or more asymmetric centers, and thus may produce diastereomers and optical isomers. Unless otherwise indicated, the present invention includes all possible diastereomers and racemic mixtures thereof, their substantially pure resolved enantiomers, all possible geometric isomers and pharmaceutically acceptable salts thereof. Also included are mixtures of stereoisomers and isolated specific stereoisomers. In the synthetic process for preparing such compounds, or in the use of racemization or epimerization procedures known to those skilled in the art, the product of such procedures may be a mixture of stereoisomers.

[0095] Many organic compounds exist in optically active forms, which have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule around its chiral center. The prefixes d and l or (+) and (-) are used to indicate the sign of rotation of plane polarized light of the compound, (-) or l indicates that the compound is left-handed. Compounds prefixed with (+) or d are right-handed. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are non-superimposable mirror images of each other. Specific stereoisomers may also be called enantiomers, and mixtures of such isomers are often called mixtures of isomers. A 50:50 mixture of enantiomers is called a racemic mixture. Many of the compounds described herein may have one or more chiral centers and may therefore exist in different enantiomeric forms. If desired, a chiral carbon may be represented by an asterisk (*). When the bonds to a chiral carbon are depicted as straight lines in a disclosed formula, it is understood that both (R) and (S) configurations of the chiral carbon are included, and thus enantiomers and mixtures thereof are included in the formula. As used in the art, when it is necessary to specify the absolute configuration of a chiral carbon, one bond to the chiral carbon may be represented as a wedge (bonded to an atom above the plane) and the other may be represented as a set of short parallel lines or a wedge of short parallel lines (bonded to an atom below the plane). The Cahn-Ingold-Prelog system may be used to assign the (R) or (S) configuration to a chiral carbon.

[0096] The compounds described herein include atoms of natural isotopic abundance and non-natural abundance. The disclosed compounds may be isotopically labeled or isotopically substituted compounds identical to the compounds described, but one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or atomic mass number usually found in nature. Examples of isotopes that may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, and chlorine, such as 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 35 S. 18 F and 36 Cl. Compounds also include prodrugs thereof and pharmaceutically acceptable salts of the compounds or prodrugs, which contain the above-mentioned isotopes and / or other isotopes of other atoms, all of which are within the scope of the present invention. Certain isotope-labeled compounds of the present invention, for example, those in which radioactive isotopes such as 3 H and 14 C compounds can be used for drug and / or substrate tissue distribution assays. Tritium isotopes, i.e. 3 H, and carbon-14 isotopes, i.e.14 C is particularly preferred because it is easy to prepare and detect. In addition, the use of heavier isotopes such as deuterium (i.e. 2 H) substitution may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in certain circumstances. Isotopically labeled compounds of the invention and their prodrugs can generally be prepared by the following procedure, i.e., substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.

[0097] The compounds described in the present invention can exist in the form of solvates. In some cases, the solvent used to prepare the solvate is an aqueous solution, and the solvate is generally referred to as a hydrate. These compounds can exist in the form of a hydrate, for example, can be obtained by crystallization from a solvent or an aqueous solution. In this regard, one, two, three or any number of solvents or water molecules can be combined with the compounds of the present invention to form solvates and hydrates. Unless otherwise indicated, the present invention includes all possible solvates.

[0098] It should also be appreciated that certain compounds described herein may exist as an equilibrium of tautomers. For example, a ketone with an α-hydrogen may exist in an equilibrium of a keto form and an enol form.

[0099]

[0100] Likewise, amides having an N-hydrogen may exist in equilibrium between the amide form and the imidic acid form. Unless otherwise specified, the present invention includes all possible tautomers.

[0101] It is well known that chemical substances form solids that exist in different ordered states, called polymorphs or modifications. Different modifications of polymorphic substances may have very different physical properties. The compounds of the present invention may exist in different polymorphs, wherein a specific modification may be metastable. Unless otherwise stated, the present invention includes all possible polymorphs.

[0102] In some aspects, the structure of the compound can be represented by the following formula:

[0103]

[0104] It is understood to be equivalent to the following formula:

[0105]

[0106] where n is usually an integer. That is, R n is understood to mean five independent substituents R n(a) , R n(b) , R n(c) , Rn(d) and R n(e) "Independent substituents" means that each R substituent can be defined independently. For example, if in one example R n(a) is a halogen, then R n(b) Not necessarily a halogen in this example.

[0107] As used herein, "administering" may refer to oral, topical, intravenous, subcutaneous, transdermal, transdermal, intramuscular, intraarticular, parenteral, intraarteriole, intradermal, intraventricular, intraosseous, intraocular, intracranial, intraperitoneal, intralesional, intranasal, intracardiac, intraarticular, intracavernous, intrathecal, intravenous, intracerebral and intraventricular, intratympanic, intracochlear, rectal, vaginal, active or passive (e.g., by diffusion) administration by inhalation, catheter, stent, or by implanted reservoir or other device, introducing the composition into the perivascular space and adventitia. For example, a medical device such as a stent may contain a composition or formulation placed on its surface, which may then dissolve or otherwise distribute into surrounding tissues and cells. The term "parenteral" may include subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Administration may be continuous or intermittent. In various aspects, the formulation may be administered therapeutically; i.e., for the treatment of an existing disease or condition. In further various aspects, the formulations can be administered prophylactically; that is, to prevent a disease or condition.

[0108] As used interchangeably herein, "subject", "individual" or "patient" may refer to a vertebrate, such as a mammal (e.g., a human). "Subject" may also refer to a cell, a cell population, a tissue, an organ or an organism, preferably a human and its parts.

[0109] As used herein, the terms "treating" and "treatment" may generally refer to obtaining a desired pharmacological and / or physiological effect. The effect may be, but need not necessarily be, preventive in terms of preventing or partially preventing a disease, symptom or condition (e.g., hematological malignancies, breast cancer, and / or other solid malignancies). The effect may be therapeutic in terms of partial or complete cure of a disease, condition, symptom, or adverse reaction attributed to the disease, disorder, or condition. The term "treatment" as used herein may include any treatment of a subject, particularly a hematological malignancy, cancer breast cancer, and / or other solid tumors in a human, and may include any one or more of the following: (a) preventing the disease from occurring in a subject who may be susceptible to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, i.e., preventing its development; and (c) alleviating the disease, i.e., alleviating or improving the disease and / or its symptoms or conditions. The term "treatment" as used herein may refer to a single treatment, a single preventive treatment, or a therapeutic and preventive treatment. A person in need of treatment (a subject in need of treatment) may include those who already have the disease and / or need to prevent the disease. As used herein, the term "treating" can include inhibiting a disease, disorder, or condition, e.g., hindering its progression; as well as alleviating a disease, disorder, or condition, e.g., causing regression of the disease, disorder, and / or condition. Treating a disease, disorder, or condition can include ameliorating at least one symptom of a particular disease, disorder, or condition, even if the underlying pathophysiology is not affected, e.g., treating pain in a subject by administering an analgesic, even if such drug does not treat the cause of the pain.

[0110] As used herein, "therapeutic" may refer to treating, curing, and / or ameliorating a disease, disorder, condition, or side effect, or reducing the rate of progression of a disease, disorder, or side effect.

[0111] As used herein, an "effective amount" may refer to an amount of a disclosed compound or pharmaceutical composition provided herein that is sufficient to affect a beneficial or desired biological, affective, medical, or clinical response in a cell, tissue, system, animal, or human. An effective amount may be administered in one or more administrations, applications, or dosages. The term may also include within its scope an amount that is effective to enhance or restore substantially normal physiological function.

[0112] For example, those skilled in the art are fully capable of starting to administer the compound at a level lower than that required to achieve the desired therapeutic effect, and gradually increasing the dosage until the desired effect is achieved. If necessary, for the purpose of administration, the effective daily dose can be divided into multiple doses. Therefore, a single-dose composition can contain such an amount or its sub-multiples to constitute a daily dose. If there are any contraindications, the dosage can be adjusted by an individual doctor. It is generally preferred to use the maximum dose of the pharmacological agent of the present invention (alone or in combination with other therapeutic agents), i.e., the highest safe dose according to reasonable medical judgment. However, it will be understood by those of ordinary skill in the art that patients may adhere to a lower dose or a tolerable dose for medical reasons, psychological reasons, or almost any other reason.

[0113] For example, the response to a therapeutically effective dose of the disclosed compounds and / or pharmaceutical compositions can be measured by determining the physiological effects of the treatment or drug, such as the reduction or disappearance of disease symptoms after administration of the therapeutic or pharmacological agent. Other assays will be known to those of ordinary skill in the art and can be used to measure the level of response. The amount of treatment can be varied, for example, by increasing or decreasing the amount of the disclosed compounds and / or pharmaceutical compositions, by changing the disclosed compounds and / or pharmaceutical compositions administered, by changing the route of administration, by changing the dosage time, etc. The dosage can be varied and can be administered once or more times per day for one day or several days. Appropriate dosage guidelines for a given class of drugs can be found in the literature.

[0114] As used herein, the term "prophylactically effective amount" refers to an amount effective to prevent the onset or initiation of a disease or disorder.

[0115] As used herein, the term "prevent" or "preventing" means to exclude, avoid, eliminate, prevent, stop or hinder the occurrence of something, especially by preemptive action. It should be understood that unless otherwise specifically stated, when reducing, inhibiting or preventing is used in this article, the use of the other two words is also explicitly disclosed.

[0116] The term "pharmaceutically acceptable" describes a material that is biologically or otherwise undesirable, ie, does not cause unacceptable adverse biological effects or interact in a deleterious manner.

[0117] The term "pharmaceutically acceptable salt" as used herein refers to a salt of an active agent prepared with an acid or base that is tolerated by a biological system or a subject or that is tolerated by a biological system and that is tolerated by a subject when administered in an effective therapeutic amount. When the compounds of the present disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, whether pure alkali or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, ammonium, organic amino, magnesium salts, lithium salts, strontium salts or similar salts. When the compounds of the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, which can be pure or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include, but are not limited to, salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and salts derived from relatively nontoxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are amino acid salts such as arginine salts, and organic acid salts such as glucuronic acid or galacturonic acid, etc.

[0118] The term "pharmaceutically acceptable prodrug" or "prodrug" means a prodrug of the disclosed compounds that is suitable for use in contact with the tissues of humans and lower animals, without excessive toxicity, irritation, allergic reaction, etc., within the scope of reasonable medical judgment, has a reasonable benefit / risk ratio, and is effective for its intended use. The prodrugs disclosed herein can be rapidly converted in vivo to the parent compound having the structure of the disclosed compound, for example, by hydrolysis in the blood. T. Higuchi and V. Stella (Pro-drugs as Novel Delivery Systems, V. 14 of the ACSSymposium Series) and Edward B. Roche (ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987)) are discussed in depth.

[0119] As used herein, "dose", "unit dose" or "dosage" may refer to physically discrete units suitable for use in subjects, each unit containing a predetermined quantity of a disclosed compound and / or pharmaceutical composition thereof calculated to produce a desired response or the desired response associated with its administration.

[0120] Certain materials, compounds, compositions and components disclosed herein can be obtained commercially or readily synthesized by techniques generally known to those skilled in the art. For example, the starting materials and reagents used to prepare the disclosed compounds and compositions can be obtained from commercial suppliers such as Aldrich Chemical Co. (Milwaukee, Wisconsin), Acros Organics (Morris Plains, New Jersey), Fisher Scientific (Pittsburgh, Pennsylvania), or Sigma (St. Louis, Missouri), or prepared by methods known to those skilled in the art according to procedures set forth in references (e.g., Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition) and Larock's Comprehensive Organic Transformations (VCH Publishers, 1991). Inc.,1989)).

[0121] Unless explicitly stated otherwise, any method described herein should not be construed as requiring that its steps be performed in a specific order. Therefore, if a method claim does not actually list the order in which its steps should be followed, or if the claim or specification does not specifically state that the steps should be limited to a specific order, then no order is meant to be inferred in any way. This applies to any possible non-express basis for interpretation, including: logical issues related to the arrangement of steps or operational flow; plain meaning derived from grammatical organization or punctuation; and the number or type of embodiments described in the specification.

[0122] Components for preparing the compositions of the present invention and the compositions themselves used in the methods disclosed herein are disclosed. These and other materials are disclosed herein, and it should be understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed, although specific references to various individual and collective combinations and permutations of these compounds cannot be explicitly disclosed, each combination and permutation is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed, and multiple modifications that can be made to multiple molecules including the compound are discussed, each combination and permutation of the compound and possible modifications are specifically contemplated, unless otherwise specifically stated. Thus, if a class of molecules A, B, and C and a class of molecules D, E, and F and an example of a combined molecule are disclosed, AD is disclosed, then even if each is not listed individually, each can be considered individually and collectively, which means that the combinations AE, AF, BD, BE, BF, CD, CE, and CF are also considered to be disclosed. Similarly, any subset or combination of these is also disclosed. Thus, for example, a subgroup of AE, BF, and CE will be considered to be disclosed. This concept applies to all aspects of the present application, including but not limited to steps in the method of preparing and using the compositions of the present invention. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the methods of the invention.

[0123] It is understood that the compositions disclosed herein have certain functions. Certain structural requirements for performing the disclosed functions are disclosed herein, and it is understood that there are multiple structures that can perform the same functions associated with the disclosed structures, and these structures will generally achieve the same results.

[0124] As used herein, the terms "optional" or "optionally" means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0125] Unless otherwise specified, temperatures referred to herein are based on atmospheric pressure (ie, one atmosphere).

[0126] Compounds and methods of making and using the same

[0127] In one aspect, disclosed herein is a compound having a structure of Structure I or a pharmaceutically acceptable salt thereof

[0128]

[0129] in

[0130] R 1is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0131] n is an integer from 1 to 5, wherein each R 2 are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, halide or alkoxy;

[0132] m is an integer from 1 to 3, wherein each R 3 are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, halide or alkoxy;

[0133] o is an integer from 1 to 10;

[0134] X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0135] Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0136] R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0137] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0138] Each Z is independently hydrogen or deuterium.

[0139] In one aspect, X in Structure I is O. In another aspect, Y in Structure I is O. In another aspect, Y in Structure I is NR 5 , where R 5is a C1 to C5 alkyl group. In another aspect, Y in structure I is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group. In another aspect, R in structure I 1 is hydrogen. On the other hand, R in structure I 3 On the other hand, R in structure I 3 is an alkoxy group, and m in Structure I is 1. In another aspect, o in Structure I is an integer from 1 to 5. In another aspect, R in Structure I 2 is halogen, and n is 2. In another aspect, the ortho-positioned R 2 is a fluoride. On the other hand, R in structure I 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h On the other hand, R in structure I 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h They are deuterium.

[0140] In another aspect, the compound has structure II or a pharmaceutically acceptable salt thereof

[0141]

[0142] in

[0143] R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0144] R 2a and R 2b It is a halide;

[0145] R 3 is an alkoxy group;

[0146] o is an integer from 1 to 5;

[0147] X is O or NR 4 , where R 4is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0148] Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0149] R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0150] R 7a 、R7 b , R 7c , R 7d 、R7 e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0151] Each Z is independently hydrogen or deuterium.

[0152] In one aspect, X in Structure II is O. In another aspect, Y in Structure II is O. In another aspect, Y in Structure II is NR 5 , where R 5 is C1 to C5 alkyl. In another aspect, Y in structure II is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group. In another aspect, R in Structure II 1 In another aspect, R 3 is a C1 to C10 substituted or unsubstituted straight or branched chain alkoxy. In another aspect, R in structure II 3 In another aspect, o in Structure II is an integer from 1 to 5. In another aspect, R in Structure II 2 is a halide. In another aspect, R in Structure II 2a is chloride, R 2b is a fluoride. In another aspect, R in Structure II 2a is fluoride, R 2b is chloride. In another aspect, R in Structure II7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h In another aspect, R in Structure II 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h They are deuterium.

[0153] In another aspect, the compound has structure III or a pharmaceutically acceptable salt thereof

[0154]

[0155] in

[0156] R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0157] R 2a and R 2b It is a halide;

[0158] R 3 is an alkoxy group;

[0159] o is an integer from 1 to 5;

[0160] X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0161] Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0162] R 6a and R 6b are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0163] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0164] Each Z is independently hydrogen or deuterium.

[0165] In one aspect, X in Structure III is O. In another aspect, Y in Structure III is O. In another aspect, Y in Structure III is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group. In another aspect, R in structure III 1 is hydrogen. In another aspect, R in Structure III 3 is a C1 to C10 substituted or unsubstituted straight or branched chain alkoxy. In another aspect, R in structure III 3 In another aspect, o in Structure III is an integer from 1 to 5. In another aspect, R in Structure III 2 is a halide. In another aspect, R in Structure III 2a is chloride, R 2b is a fluoride. In another aspect, R in Structure III 2a is fluoride, R 2b is chloride. In another aspect, R in Structure II 2a is fluoride, R 2b is chloride. In another aspect, R in Structure III 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h In another aspect, R in Structure III 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h They are deuterium.

[0166] In another aspect, the compound has the structure

[0167]

[0168] General Synthesis Methods

[0169] In one aspect, the compounds described herein can be prepared by reacting a compound having structure IV with a compound having structure V in the presence of a base.

[0170]

[0171] wherein the variables in structures IV and V are as described above and LG is a leaving group.

[0172] The reaction between compounds having structures IV and V is typically carried out in an organic solvent, wherein an appropriate amount of a base is provided to deprotonate the XH proton in structure IV. In one aspect, the base comprises a hydride, an alkoxide, a Grignard reagent, or an alkyl lithium compound. In one aspect, the leaving group LG in structure V is a chloride or sulfonic acid group. In one aspect, the compounds described herein can be used Figure 4-6 and Figure 8 The procedures provided in are produced. Compounds having structures IV and V can be synthesized using organic techniques or purchased. In one aspect, the compound having structure IV is CAS No. 184475-71-6.

[0173] Also described herein are synthetic methods for large-scale, high-yield production of piperazinyl and morpholinohydroxylamine. The synthetic methods require minimal steps and can produce piperazinyl and morpholinohydroxylamine on a multi-gram scale. Piperazinyl and morpholinohydroxylamine are useful intermediates for producing the compounds described herein.

[0174] Figure 3 and Figure 17-18 Exemplary general synthetic methods for preparing piperazinyl and morpholinohydroxylamines are provided.

[0175] Reference Fig.17 , using four steps AD to prepare piperazinylhydroxylamine with structure X

[0176]

[0177] in

[0178] R 5 is a substituted or unsubstituted straight chain or branched chain alkyl group, and

[0179] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0180] each Z is independently hydrogen or deuterium,

[0181] In one aspect, a method for preparing a piperazinylhydroxylamine having structure X comprises

[0182] (A) reacting a compound having structure XI with a compound having structure XII in the presence of a base to produce a compound having structure XIII

[0183]

[0184] Where R 10 is a substituted or unsubstituted straight chain or branched chain alkyl group, or a substituted or unsubstituted straight chain or branched chain alkoxy group, and

[0185] LG is the leaving group;

[0186] (B) reacting a compound having structure XIII with a first oxidant under heating to produce a compound having structure XIV

[0187]

[0188] (C) reacting a compound having structure XIV with (i) a second oxidizing agent and then with (ii) a first reducing agent to produce a first intermediate; and

[0189] (D) reacting the first intermediate with a second reducing agent to produce a compound having structure X.

[0190] Step A involves reacting a compound having structure XI with a compound having structure XII in the presence of a base to prepare a compound having structure XIII

[0191]

[0192] in

[0193] R 5 is a substituted or unsubstituted straight chain or branched chain alkyl group, and

[0194] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl;

[0195] Each Z is independently hydrogen or deuterium;

[0196] R 10 is a substituted or unsubstituted straight chain or branched chain alkyl group, or a substituted or unsubstituted straight chain or branched chain alkoxy group, and

[0197] LG is the leaving group;

[0198] In one aspect, R in structures XI and XIII 10 is a C1-C5 straight or branched alkyl group. In another aspect, R in structures XI and XIII 10 is a C1-C5 straight or branched chain alkoxy. In another aspect, R in structures XI and XIII 10 is a C1-C5 straight or branched alkoxy group substituted with an aryl group. In another aspect, R in structures XI and XIII 10 It is benzyloxy.

[0199] Step A relates to the allylation of a compound with structure XI and an allyl compound with structure XII in the presence of a base. In one aspect, the leaving group LG of the allyl compound XII is a halide, a sulfonate, a carbonate or a phosphate. In another aspect, the leaving group is a bromide. In one aspect, the allyl compound XII can be partially or completely deuterated. In another aspect, each Z in the allyl compound XII is hydrogen. The molar ratio of the compound with structure XI to the allyl compound with structure XII can be 0.5:1 to 1:3 or 0.5:1, 1:1, 1:1.5, 1:2, 1:2.3 or 1:3, wherein any value can be the lower limit or upper limit of the range (e.g., 1:1.5 to 1:2).

[0200] Step A is carried out in an organic solvent. In one aspect, the organic solvent is an aprotic organic solvent, such as tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-methylTHF), diethyl ether, methyl tert-butyl ether (MTBE), 1,4-dioxane, 1,2-dimethoxyethane, pentane, hexane, heptane, cyclohexane, N, N'-dimethylpropylene urea (DMPU) or a combination thereof.

[0201] Base is a compound that can deprotonate the amino proton on the piperazine ring of compound XI. In one aspect, it includes carbonates, hydroxides, phosphates, hydrides, dialkylamides or hexamethyldisilazane. The molar ratio of the compound having structure XI to the base can be 0.5:1 to 1:3, or 0.5:1, 1:1, 1:1.5, 1:2, 1:2.3 or 1:3, wherein any value can be the lower or upper limit of the range (e.g., 1:1.5 to 1:2).

[0202] In one aspect, step A is carried out at an elevated temperature. In one aspect, the reaction in step A is carried out at a temperature of about 25°C to about 100°C or 25°C, 35°C, 45°C, 55°C, 65°C, 75°C, 85°C, 95°C or 100°C, any of which may be the lower or upper limit of the range (e.g., 55°C to 75°C). After step A, a compound of structure XIV is prepared. The compound of structure XIII may then be purified using techniques known in the art.

[0203] Step B involves oxidation of a compound having structure XIII followed by a Meisenheimer rearrangement to produce a compound having structure XIV.

[0204] In one aspect, step B comprises the following steps:

[0205] (i) reacting a compound having structure XIII with a first oxidizing agent in a first organic solvent to prepare a first composition;

[0206] (ii) adding an aqueous base to the first composition to prepare a second composition comprising an organic layer and an aqueous layer;

[0207] (iii) separating the organic layer from the aqueous layer;

[0208] (iv) removing the first organic solvent from the organic layer to produce a residue;

[0209] (v) dissolving the residue in a second organic solvent to prepare a second composition; and

[0210] (vi) heating the second composition from about 50°C to about 100°C to produce a compound having structure XIV.

[0211] In one aspect, the first oxidant in step B comprises a peroxyacid, ozone (oxone) or hydrogen peroxide / acetic acid. In another aspect, the first oxidant in step B comprises meta-chloroperbenzoic acid. In another aspect, the molar ratio of the first oxidant to the compound of structure XIII is 0.95:1 to 1:1.05.

[0212] Step B is carried out in an organic solvent, wherein the solvent does not react with the first oxidant. In one aspect, the first organic solvent is dichloromethane. The first solvent can then be removed using techniques known in the art, and the remaining residue is then dissolved in a second organic solvent to prepare a second composition. In one aspect, the second organic solvent is a high boiling point solvent, such as toluene. In one aspect, the second composition in step B is reacted at a temperature of about 50°C to about 100°C or 50°C, 60°C, 70°C, 80°C, 90°C or 100°C, wherein any value can be the lower or upper limit of the range (e.g., 70°C to 90°C). In the heating step, a Meisenheimer rearrangement occurs to produce a compound with structure XIV. The compound with structure XIIV can then be purified using techniques known in the art.

[0213] Step C involves oxidation of the olefinic group in the compound having structure XIIV followed by reduction to produce the first intermediate.

[0214] In one aspect, step C comprises the following steps:

[0215] (i) reacting a compound having structure XIV with a second oxidizing agent in a third organic solvent to produce a third composition; and

[0216] (ii) mixing a first reducing agent with a third composition to prepare a first intermediate.

[0217] In one aspect, the second oxidant used in step C comprises ozone or osmium tetroxide and sodium periodate. Step C is carried out in an organic solvent. In one aspect, the third organic solvent comprises an alcohol (e.g., methanol, ethanol, butanol, or any combination thereof) and an aprotic solvent (e.g., tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-methylTHF), ether, methyl tert-butyl ether (MTBE), 1,4-dioxane, 1,2-dimethoxyethane, pentane, hexane, heptane, cyclohexane, N,N′-dimethylpropylene urea (DMPU), or any combination thereof).

[0218] In one aspect, the compound having structure XIV is reacted with a second oxidant at a temperature of about -50°C to about -100°C, or -50°C, -60°C, -70°C, -80°C, 90°C, or -100°C, any of which may be the lower or upper limit of the range (e.g., -70°C to -90°C).

[0219] After oxidation of the olefinic group in the compound having structure XIIV, a first reducing agent is added to the reaction to produce a first intermediate which is a terminal alcohol provided as follows

[0220]

[0221] In one aspect, the first reducing agent comprises a hydride, which is any compound capable of delivering a hydride ion. In one aspect, the first reducing agent comprises a borohydride. In one aspect, the hydride is Sodium borohydride , Lithium aluminum hydride or Hydrogenated diisocyanate Butyl aluminum The molar ratio of the first reducing agent to the compound having structure XIV is 1.5:1 to 2.5:1, or 1.5:1, 1.75:1, 2:1, 2.25:1, or 2.5:1, wherein any value can be the lower or upper limit of the range (e.g., 1.75:1 to 2:1). The first intermediate produced in step C can then be purified prior to step D using techniques known in the art.

[0222] Step D involves reducing the carbonyl group in the first intermediate to prepare the piperazinylhydroxylamine compound X.

[0223] In one aspect, step D comprises the following steps:

[0224] (i) dissolving the first intermediate in an aprotic solvent to prepare a fourth composition;

[0225] (ii) mixing a second reducing agent with the fourth composition to produce a compound having structure X; and

[0226] (iii) isolating and purifying the compound having structure X.

[0227] The first intermediate is dissolved in an aprotic solvent (e.g., tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-methylTHF), diethyl ether, methyl tert-butyl ether (MTBE), 1,4-dioxane, 1,2-dimethoxyethane, pentane, hexane, heptane, cyclohexane, N,N'-dimethylpropylene urea (DMPU), or any combination thereof), and then a second reducing agent is added. In one aspect, the hydride is Sodium borohydride , Lithium aluminum hydride or Diisobutylaluminum hydride The molar ratio of the first reducing agent to the compound having structure XIV is 2:1 to 4:1, or 2:1, 2.5:1, 3:1, 3.5:1, or 4:1, any of which may be the lower or upper limit of the range (e.g., 2.5:1 to 3.5:1).

[0228] In one aspect, the first intermediate is reacted with the second reducing agent at a temperature of about 10°C to about -50°C, or 10°C, 0°C, -10°C, -20°C, -30°C, -40°C, or -50°C, wherein any value can be the lower or upper limit of a range (e.g., -10°C to -30°C). The piperazinylhydroxylamine compound X produced in step D can then be purified prior to step D using techniques known in the art.

[0229] Reference Fig.18 , using three steps AC to prepare morpholinohydroxylamine with structure XX

[0230]

[0231] in

[0232] R 7a , R 7b 、R7 c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0233] each Z is independently hydrogen or deuterium,

[0234] In one aspect, a method for preparing a morpholinohydroxylamine having structure XX comprises

[0235] (A) reacting a compound having structure XXI with a compound having structure XXII in the presence of a base to prepare a compound having structure XXIII

[0236]

[0237] Where LG is the leaving group;

[0238] (B) reacting a compound having structure XXIII with a first oxidant to produce a compound having structure XXIV as well as

[0239] (C) reacting a compound having structure XXIV with (i) a second oxidizing agent and then with (ii) a first reducing agent to produce a compound having structure XX.

[0240] The same reaction conditions and reagents used to produce piperazinylhydroxylamine in Steps AC can be used to produce morpholinohydroxylamine having structure XX, wherein Step D is not required to produce the morpholinohydroxylamine.

[0241] Exemplary methods for preparing the compounds described herein, as well as characterization information, are provided in the Examples. Solvents, temperature, the presence or absence of protecting groups, and other reaction conditions may vary depending on the specific substituents in the compound being synthesized.

[0242] Exemplary methods for preparing the compounds described herein, as well as characterization information, are provided in the Examples. Solvents, temperature, the presence or absence of protecting groups, and other reaction conditions may vary depending on the specific substituents in the compound being synthesized.

[0243] Pharmaceutical composition

[0244] In various aspects, the present disclosure relates to pharmaceutical compositions comprising an effective therapeutic amount of at least one disclosed compound, at least one product of a disclosed method, or a pharmaceutically acceptable salt thereof. As used herein, "pharmaceutically acceptable carrier" refers to one or more pharmaceutically acceptable diluents, preservatives, antioxidants, solubilizers, emulsifiers, colorants, release agents, coating agents, sweeteners, flavoring agents and spices, and adjuvants. The disclosed pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any method well known in the art of pharmacy and pharmaceutical science.

[0245] In another aspect, the disclosed pharmaceutical composition comprises an effective therapeutic amount of at least one disclosed compound, at least one product of the disclosed method, or a pharmaceutically acceptable salt thereof as an active ingredient, a pharmaceutically acceptable carrier, optionally one or more other therapeutic agents, and optionally one or more adjuvants. The disclosed pharmaceutical composition includes pharmaceutical compositions suitable for oral, rectal, topical, pulmonary, nasal, and parenteral administration, although the most suitable approach in any given case will depend on the nature and severity of the disease to which the active ingredient is administered. In another aspect, the disclosed pharmaceutical composition can be formulated to allow oral, nasal, inhaled, parenteral, paracancer, transmucosal, transdermal, intramuscular, intravenous, intradermal, subcutaneous, intraperitoneal, intraventricular, intracranial, and intratumoral administration.

[0246] As used herein, "parenteral administration" includes administration by bolus injection or infusion, as well as administration by intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular subarachnoid, intraspinal, epidural, and intrasternal injection and infusion.

[0247] In various aspects, the present invention also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent, and an effective therapeutic amount of the disclosed compound, the product of the disclosed preparation method, a pharmaceutically acceptable salt, a hydrate, a solvate, a polymorph or a stereochemical isomeric form thereof as an active ingredient. In another aspect, the disclosed compound, the product of the disclosed preparation method, a pharmaceutically acceptable salt, a hydrate, a solvate, a polymorph or a stereochemical isomeric form thereof, or any subgroup or combination thereof, can be formulated into various pharmaceutical forms for administration purposes.

[0248] In practice, according to conventional pharmaceutical formulation techniques, the compounds of the present invention or their pharmaceutically acceptable salts can be closely mixed with pharmaceutical carriers as active ingredients. The carrier can take a variety of forms, depending on the desired form of administration, such as oral or parenteral administration (including intravenous injection). Therefore, the pharmaceutical composition of the present invention can be presented in the form of discrete units suitable for oral administration, such as capsules, cachets or tablets, each containing a predetermined amount of active ingredients. In addition, the composition can exist in the form of powders, granules, solutions, aqueous liquid suspensions, non-aqueous liquids, oil-in-water emulsions or water-in-oil emulsions. In addition to the above-mentioned common dosage forms, the compounds of the present invention and / or their pharmaceutically acceptable salts can also be administered by controlled release devices and / or delivery devices. The composition can be prepared by any pharmaceutical method. In general, this method includes the step of combining the active ingredient with a carrier constituting one or more essential ingredients. Typically, the composition is prepared by uniformly and closely mixing the active ingredient with a liquid carrier or a finely divided solid carrier or both. Then, the product can be conveniently formed into the desired appearance.

[0249] For ease of administration and dosage uniformity, it is particularly advantageous to formulate the above-mentioned pharmaceutical composition in unit dosage form. The term "unit dosage form" as used herein refers to a physically discrete unit suitable as a unit dose, each unit containing a predetermined amount of active ingredient, which is calculated to produce the desired therapeutic effect in combination with the required pharmaceutical carrier. In other words, "unit dosage form" refers to a single dose in which all active and inactive ingredients are combined in a suitable system so that the patient or the person administering the drug to the patient can open a container or package containing the entire dose without mixing any of the ingredients in two or more containers or packages together. Typical examples of unit dosage forms are tablets (including scored or coated tablets), capsules or oral pills, single-dose vials for injectable solutions or suspensions, rectal suppositories, powder packs, film tablets, and separate multiples thereof. This list of unit dosage forms is not intended to be limiting in any way, but only represents typical examples of unit dosage forms.

[0250] Pharmaceutical compositions disclosed herein include compounds of the present disclosure (or their pharmaceutically acceptable salts) as active ingredients, pharmaceutically acceptable carriers and optionally one or more other therapeutic agents. In various aspects, the disclosed pharmaceutical compositions may include pharmaceutically acceptable carriers and disclosed compounds or their pharmaceutically acceptable salts. In another aspect, the disclosed compounds or their pharmaceutically acceptable salts may also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds. Example compositions include compositions suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most appropriate approach in any particular case will depend on the nature and severity of the disease for which the active ingredient is administered. Pharmaceutical compositions may be conveniently presented in unit dosage form and prepared by any method known in the pharmaceutical field.

[0251] For example, techniques and compositions for preparing dosage forms useful for the materials and methods described herein are described in the following references: Modern Pharmaceutics, Chapters 9 and 10 (Banker and Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Eds., 1996); McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; JG Hardy, SSDavis, CliveG.Wilson, Eds.); Modern Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol40 (Gilbert S.Banker, Christopher T. Rhodes, Eds.). .

[0252] Compounds as described herein are usually administered in combination with suitable pharmaceutical diluents, excipients, fillers or carriers (referred to herein as pharmaceutically acceptable carriers or carriers), which are appropriately selected according to the expected form of administration and in accordance with conventional pharmaceutical practice. Deliverable compounds will be in a form suitable for oral, rectal, local, intravenous or parenteral administration. The carrier includes solids or liquids, and the type of carrier is selected according to the type of administration used. The compound can be administered in a dosage with a known amount of the compound.

[0253] Oral administration may be the preferred dosage form due to ease of administration, with tablets and capsules being the most advantageous oral dosage unit forms, in which case solid pharmaceutical carriers are obviously used. However, other dosage forms may be suitable depending on the clinical population (e.g., age and severity of the clinical condition), the solubility of the specific disclosed compound used, etc. Therefore, the disclosed compounds can be used in oral dosage forms such as pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. When preparing a composition of an oral dosage form, any convenient pharmaceutical medium can be used. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, colorants, etc. can be used to form oral liquid preparations such as suspensions, elixirs and solutions; and carriers such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc. can be used to form oral solid preparations such as powders, capsules and tablets. Due to their ease of administration, tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers. Optionally, tablets can be coated by standard aqueous or non-aqueous techniques.

[0254] The disclosed pharmaceutical composition of oral dosage form may contain one or more pharmaceutical excipients and / or additives. Non-limiting examples of suitable excipients and additives include gelatin, natural sugars such as raw sugar or lactose, lecithin, pectin, starch (such as corn starch or amylose), dextran, polyvinyl pyrrolidone, polyvinyl acetate, gum arabic, alginic acid, tylose, talc, lycopodium, silica gel (such as colloid), cellulose, cellulose derivatives (such as cellulose ethers partially etherified with low-saturated fatty alcohols and / or low-saturated fatty oxygen alcohols, such as methyloxypropylcellulose, methylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethyl phthalate), fatty acids and magnesium, calcium or aluminum salts of fatty acids with 12 to 22 carbon atoms, especially saturated (such as stearates), emulsifiers, oils and fats, especially from vegetables (such as peanut oil, castor oil, olive oil, sesame oil, cottonseed oil, corn oil, wheat germ oil, sunflower seed oil, cod liver oil, in each case also optionally hydrated); saturated fatty acids C 12 H 24O2 to C 18 H 36 O2 glycerol esters and polyglycerol esters and mixtures thereof, the glycerol hydroxyl groups can be completely or only partially esterified (for example monoglyceride, diglyceride and triglyceride); pharmaceutically acceptable monovalent alcohols or polyvalent alcohols and polyethylene glycols, such as polyethylene glycol and its derivatives, aliphatic saturated or unsaturated fatty acids (2 to 22 carbon atoms, especially 10-18 carbon atoms) with monovalent fatty alcohols (1 to 20 carbon atoms) or polyvalent alcohols such as ethylene glycol, glycerol, diethylene glycol, pentaerythritol, sorbitol, mannitol, etc., which can also be optionally etherified, esters of citric acid with primary alcohols, acetic acid, urea, benzyl benzoate, dioxolane, glycerol formaldehyde, tetrahydrofurfuryl alcohol, polyethylene glycol ethers with C1-C12 alcohols, dimethylacetamide, lactams, lactates, ethyl carbonate, silicones (especially medium viscosity polydimethylsiloxane), calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate, magnesium carbonate, etc.

[0255] Other auxiliary substances which can be used for the preparation of oral dosage forms are those which cause disintegration (so-called disintegrants), such as cross-linked polyvinyl pyrrolidone, sodium carboxymethyl starch, sodium carboxymethyl cellulose or microcrystalline cellulose. Conventional coating substances can also be used for the production of oral dosage forms. For example, polymers and copolymers of acrylic acid and / or methacrylic acid and / or their esters come into consideration; copolymers of acrylic acid and methacrylic acid esters with a low ammonium content (e.g. Eudragit RRS), copolymers of acrylic acid and methacrylic acid esters with trimethylammonium methacrylate (e.g. Eudragit RRS), RL); polyvinyl acetate; fats, oils, waxes, fatty alcohols; hydroxypropyl methylcellulose phthalate or acetate succinate; cellulose acetate phthalate, starch acetate phthalate and polyvinyl acetate phthalate, carboxymethyl cellulose; methylcellulose phthalate, methylcellulose succinate, phthalate succinate and methylcellulose phthalate half ester; zein; ethyl cellulose and ethylcellulose succinate; shellac, gluten; ethyl carboxyethyl cellulose; ethyl acrylate-maleic anhydride copolymer; maleic anhydride vinyl methyl ether copolymer; styrene maleic acid copolymer; 2-ethylhexyl acrylate maleic anhydride; crotonic acid-vinyl acetate copolymer; glutamic acid / glutamate copolymer; carboxymethyl ethyl cellulose glycerol monocaprylate; cellulose acetate succinate; polyarginine.

[0256] Plasticizers that can be considered as coating materials in the disclosed oral dosage forms are: citrate and tartrate esters (acetyl triethyl citrate, acetyl tributyl citrate, tributyl citrate, tributyl citrate); glycerol and glycerides (glycerol diacetate, glycerol triacetate, acetylated monoglycerides, castor oil); phthalates (dibutyl phthalate, dipentyl phthalate, diethyl phthalate, dimethyl phthalate, dipropyl phthalate), di-(2-methoxy- or 2-ethylhexyl) phthalate (di-(2-methoxy- or 2-ethylhexyl) phthalate); 2-ethoxyethyl)-adipate), ethyl glycolate, butyl phthaloyl glycolate and butyl glycolate; alcohols (propylene glycol, polyethylene glycols of various chain lengths), adipates (diethyl adipate, di(2-methoxy- or 2-ethoxyethyl) adipate); benzophenone; diethyl and dibutyl sebacic acid esters, dibutyl succinate, dibutyl tartrate; diethylene glycol dipropionate; ethylene glycol diacetate, ethylene glycol dibutyrate, ethylene glycol dipropionate; tributyl phosphate; tributyrin; polyethylene glycol sorbitan monooleate (polysorbate, such as polysorbate 50); sorbitan monooleate.

[0257] In addition, suitable binders, lubricants, disintegrants, colorants, flavoring agents, glidants and melting agents can be used as carriers. The pharmaceutical carrier adopted can be, for example, solid, liquid or gas. The example of solid carrier includes but is not limited to lactose, carclazyte, sucrose, glucose, methylcellulose, dicalcium phosphate, calcium sulfate, mannitol, sorbitol, talc, starch, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. The example of liquid carrier is syrup, peanut oil, olive oil and water. The example of gaseous carrier includes carbon dioxide and nitrogen.

[0258] In various aspects, binders can include, for example, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, etc. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, etc. In another aspect, disintegrants can include, for example, starch, methylcellulose, agar, bentonite, xanthan gum, etc.

[0259] In various aspects, oral dosage forms, such as solid dosage forms, can include a disclosed compound attached to a polymer as a targetable drug carrier or prodrug. Suitable biodegradable polymers that can be used to achieve controlled release of the drug include, for example, polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and hydrogels, preferably covalently crosslinked hydrogels.

[0260] Tablets may contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients suitable for tablet manufacture. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrants such as corn starch or alginic acid; binders such as starch, gelatin or acacia, and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period of time.

[0261] Tablets containing the disclosed compounds can be prepared by compression or molding, optionally with one or more auxiliary ingredients or adjuvants. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form (such as a powder or granules) in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, surfactant or dispersant. Molded tablets can be made by molding in a suitable machine, wetting a mixture of powdered compounds with an inert liquid diluent.

[0262] In various aspects, solid oral dosage forms, such as tablets, can be coated with enteric coatings to prevent rapid decomposition in the stomach. In various aspects, enteric coating agents include, but are not limited to, hydroxypropylmethylcellulose phthalate, methacrylic acid-methacrylate copolymers, polyvinyl acetate phthalate and cellulose acetate phthalate. Akihiko Hasegawa "Application of solid dispersions of Nifedipine with enteric coating agent to prepare a sustained-release dosage form" Chem. Pharm. Bull. 33: 1615-1619 (1985). Various enteric coating materials can be selected according to the test to achieve an enteric coating dosage form designed from scratch, so that it has a preferred combination of dissolution time, coating thickness and radial compressive strength (for example, see SCPorter et al. "The Properties of Enteric Tablet Coatings Made From Polyvinyl Acetate-phthalate and Celluloseacetate Phthalate", J. Pharm. Pharmacol. 22: 42p (1970)). In another aspect, the enteric coating agent may include hydroxypropylmethylcellulose phthalate, methacrylic acid-methacrylate copolymer, polyvinyl acetate phthalate, and cellulose acetate phthalate.

[0263] In various aspects, oral dosage forms can be solid dispersions with water-soluble or water-insoluble carriers. Examples of water-soluble or water-insoluble carriers include, but are not limited to, polyethylene glycol, polyvinyl pyrrolidone, hydroxypropyl methylcellulose, phosphatidylcholine, polyoxyethylene hydrogenated castor oil, hydroxypropyl methylcellulose phthalate, carboxymethyl ethylcellulose or hydroxypropyl methylcellulose, ethyl cellulose or stearic acid.

[0264] In various aspects, oral dosage forms can be liquid dosage forms, including those that are ingested, or dosage forms administered as mouthwashes or gargles. For example, liquid dosage forms can include aqueous suspensions containing active materials mixed with excipients suitable for making aqueous suspensions. In addition, oily suspensions can be prepared by suspending the active ingredient in vegetable oils, such as peanut oil, olive oil, sesame oil or coconut oil, or suspended in mineral oils such as liquid paraffin. Oily suspensions may also contain various excipients. Pharmaceutical compositions of the present disclosure may also be in the form of oil-in-water emulsions, which may also contain excipients, such as sweeteners and flavorings.

[0265] For the preparation of solutions or suspensions, for example, water, in particular sterile water, or physiologically acceptable organic solvents such as alcohols (ethanol, propanol, isopropanol, 1,2-propylene glycol, polyethylene glycol and its derivatives, fatty alcohols, partial glycerides), oils (e.g. peanut oil, olive oil, sesame oil, almond oil, sunflower oil, soybean oil, castor oil, neat's foot oil), paraffin, dimethyl sulfoxide, triglycerides, etc. can be used.

[0266] In the case of liquid dosage forms such as drinkable solutions, the following substances can be used as stabilizers or solubilizers: lower aliphatic monovalent alcohols and polyvalent alcohols having 2-4 carbon atoms, such as ethanol, n-propanol, glycerol, polyethylene glycols with a molecular weight between 200-600 (e.g. 1 to 40% aqueous solution), diethylene glycol monoethyl ether, 1,2-propylene glycol, organic amides (e.g. amides formed by aliphatic C1-C6 carboxylic acids with ammonia or primary, secondary or tertiary C1-C4 amines or C1-C4 hydroxylamines, such as urea, carbamates, acetamide, n-methylacetamide, N,N-diethylacetamide, N,N-dimethylacetamide), lower aliphatic amines and diamines having 2-6 carbon atoms (such as ethylenediamine, hydroxyethyltheophylline, tromethamine (e.g. 0.1 to 20% aqueous solution)), aliphatic amino acids.

[0267] In preparing the disclosed liquid dosage forms, solubilizers and emulsifiers may be included, for example, the following non-limiting examples may be used: polyvinyl pyrrolidone, sorbitan fatty acid esters (such as sorbitan trioleate), phospholipids (such as lecithin), gum arabic, gum tragacanth, polyoxyethylated sorbitan monooleate and other ethoxylated fatty acid esters of sorbitan, polyoxyethylated fats, polyoxyethylated oleotriglycerides, linolizated oleotriglycerides, polyethylene oxide condensation products of fatty alcohols, alkylphenols or fatty acids, or 1-methyl-3-(2-hydroxyethyl)imidazolone-(2). In this case, polyoxyethylation means that the substance in question contains polyoxyethylene chains, the degree of polymerization of which is generally between 2 and 40, especially between 10 and 20. Such polyoxyethylated substances can be obtained, for example, by reacting hydroxyl-containing compounds (e.g. mono- or diglycerides or unsaturated compounds, such as compounds containing oleic acid groups) with ethylene oxide (e.g. 40 mol of ethylene oxide per 1 mol of glyceride). Examples of oil triglycerides are olive oil, peanut oil, castor oil, sesame oil, cottonseed oil, corn oil. See also Dr. HP Fiedler ("Lexikon der Hillsstoffe für Pharmazie, Kostnetik und angrenzende Gebiete" 1971, pages 191-195).

[0268] In various aspects, the liquid dosage form may also contain preservatives, stabilizers, buffers, flavoring agents, sweeteners, colorants, antioxidants, and complex formers, etc. For example, complex formers that may be considered are: chelate formers, such as ethylene diamine tetraacetic acid, nitrilotriacetic acid, diethylenetriamine pentaacetic acid and salts thereof.

[0269] Liquid dosage forms may need to be stabilized, optionally with a physiologically acceptable base or buffer, within a pH range of about 6 to 9. A pH value that is as neutral or slightly alkaline as possible (up to pH 8) may be preferred.

[0270] In order to improve the solubility and / or stability of the disclosed compounds in the disclosed liquid dosage forms, parenteral injection forms or intravenous injection forms, it may be advantageous to use α-, β- or γ-cyclodextrin or derivatives thereof, particularly hydroxyalkyl-substituted cyclodextrins, such as 2-hydroxypropyl-β-cyclodextrin or sulfobutyl-β-cyclodextrin. In addition, cosolvents such as alcohols can improve the solubility or stability of the compounds of the invention in pharmaceutical compositions.

[0271] In various aspects, the disclosed liquid dosage forms, parenteral injection dosage forms or intravenous injection dosage forms can also include liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholine.

[0272] Pharmaceutical compositions of the present disclosure are suitable for injection, such as parenteral administration, such as intravenous, intramuscular or subcutaneous administration. Pharmaceutical compositions for injection can be prepared as solutions or suspensions of the active compound in water. Suitable surfactants, such as hydroxypropylcellulose, can be included. Dispersions can also be prepared in glycerol, liquid polyethylene glycol and oils containing mixtures thereof. In addition, preservatives can be added to prevent the harmful growth of microorganisms.

[0273] The pharmaceutical composition of the present disclosure suitable for parenteral administration may include a sterile aqueous solution or an oily solution, a suspension or a dispersion. In addition, the composition may be in the form of a sterile powder for the temporary preparation of such sterile injection solutions or dispersions. In some aspects, the final injection form is sterile and must be an effective liquid for use in a syringe. The pharmaceutical composition should be stable under manufacturing and storage conditions; therefore, the contamination of microorganisms such as bacteria and fungi should preferably be prevented. The carrier may be a solvent or a dispersion medium comprising, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), a vegetable oil, and a suitable mixture thereof.

[0274] For example, injectable solutions can be prepared, wherein the carrier comprises a saline solution, a glucose solution, or a mixture of a saline and glucose solution. Injectable suspensions can also be prepared, in which case appropriate liquid carriers, suspending agents, etc. can be used. In some aspects, the disclosed parenteral formulations can include about 0.01-0.1M, such as about 0.05M phosphate buffer. On the other hand, the disclosed parenteral formulations can include about 0.9% saline.

[0275] In various aspects, the disclosed parenteral pharmaceutical composition may include a pharmaceutically acceptable carrier, such as an aqueous or non-aqueous solution, a suspension, and an emulsion. Examples of non-aqueous solvents are vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include, but are not limited to, water, alcohol / aqueous solutions, emulsions, or suspensions, including saline and buffered media. Parenteral solvents may include mannitol, normal serum albumin, sodium chloride solution, Ringer's dextrose, glucose and sodium chloride, lactated Ringer's solution, and fixed oils. Intravenous injection solvents include liquid and nutritional supplements, electrolyte supplements, such as supplements based on Ringer's dextrose, etc. Preservatives and other additives may also be present, such as antibacterial agents, antioxidants, finishing agents, inert gases, etc. On the other hand, the disclosed parenteral pharmaceutical composition may include a small amount of additives, such as substances that enhance isotonicity and chemical stability, such as buffers and preservatives. Injectable pharmaceutical compositions also contemplate preparations in solid form, which are intended to be converted into preparations in liquid form shortly before use. Furthermore, other adjuvants may be added to render the formulation isotonic with the blood of the subject or patient.

[0276] In addition to the above-mentioned pharmaceutical compositions, the disclosed compounds can also be formulated into long-acting preparations. Such long-acting preparations can be administered by implantation (e.g., subcutaneous or intramuscular injection) or intramuscular injection. Therefore, for example, these compounds can be formulated with suitable polymers or hydrophobic materials (e.g., as emulsions in acceptable oils) or ion exchange resins, or as slightly soluble derivatives, for example, as slightly soluble salts.

[0277] Pharmaceutical compositions of the present disclosure can be in a form suitable for topical administration. As used herein, the phrase "topical application" refers to application to a biological surface, wherein the biological surface includes, for example, skin areas (e.g., hands, forearms, elbows, legs, faces, nails, anus and genital areas) or mucous membranes. By selecting a suitable carrier and optionally other ingredients that can be included in the composition, as described below, the composition of the present invention can be formulated into any form commonly used for topical application. Topical pharmaceutical compositions can be in the form of creams, ointments, pastes, gels, lotions, milk, suspensions, aerosols, sprays, foams, powders, pads and patches. In addition, the composition can be in a form suitable for transdermal devices. These preparations can be prepared using compounds of the present disclosure or pharmaceutically acceptable salts thereof, by conventional processing methods. For example, creams or ointments are prepared by mixing hydrophilic materials and water with compounds of about 5wt% to about 10wt%, thereby producing creams or ointments with desired consistency.

[0278] In compositions suitable for transdermal administration, the carrier optionally comprises a penetration enhancer and / or a suitable wetting agent, optionally in combination with suitable additives of any nature in small proportions which do not produce significant deleterious effects on the skin. The additives may facilitate transdermal administration and / or assist in the preparation of the desired composition. These compositions may be administered in a variety of ways, for example as a transdermal patch, as a spray drop, as an ointment.

[0279] Ointment is a semisolid preparation, usually based on vaseline or petroleum derivatives. The specific ointment base to be used is the base that provides the best delivery for the active agent selected for a given preparation, and preferably, other required characteristics (for example, emollient) are also provided. Like other carriers or solvents, ointment base should be inert, stable, non-irritating and non-sensitizing. As explained by Remington (The Science and Practice of Pharmacy, 19th Ed., Easton, Pa.: Mack Publishing Co. (1995), pp.1399-1404), ointment base can be divided into four categories: greasy base, emulsifiable base, emulsion base and water-soluble base. Greasy ointment base includes, for example, vegetable oil, animal fat and the semisolid hydrocarbon obtained from petroleum. Emulsifiable ointment base, also referred to as absorbent ointment base, has little or no water content, for example includes hydroxy stearyl sulfate, anhydrous lanolin and hydrophilic vaseline. Emulsion ointment bases are water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, including, for example, cetyl alcohol, glyceryl monostearate, lanolin and stearic acid. Preferred water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weights.

[0280] Lotions are preparations that are applied to the surface of the skin without friction. Lotions are generally liquid or semi-liquid preparations in which solid particles, including active agents, are present in water or an alcohol base. Lotions are generally preferred for treating larger body parts because more fluid compositions are easy to apply. Lotions are generally suspensions of solids, generally including liquid oil emulsions of the oil-in-water type. It is generally necessary to subdivide the insoluble materials in the lotion. Lotions generally contain suspending agents to produce better dispersions, as well as compounds that can be used to locate and keep the active agent in contact with the skin, such as methylcellulose, sodium carboxymethylcellulose, etc.

[0281] Creams are viscous liquids or semisolid emulsions that can be in oil-in-water or water-in-oil form. Cream bases are usually water-washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, also called the "inner" phase, is usually composed of petrolatum and / or a fatty alcohol such as cetyl or stearyl alcohol. The volume of the aqueous phase usually (but not necessarily) exceeds that of the oil phase and usually contains a humectant. The emulsifier in cream formulations is usually a nonionic, anionic, cationic, or amphoteric surfactant. For more information, please refer to Remington (The Science and Practice of Pharmacy, supra).

[0282] Pastes are semisolid dosage forms in which the bioactive agent is suspended in a suitable matrix. Depending on the nature of the matrix, pastes are classified as fat pastes or pastes made from a single-phase hydrogel. The matrix in a fat paste is usually vaseline, hydrophilic vaseline, etc. Pastes made from a single-phase hydrogel usually contain carboxymethylcellulose, etc. as a matrix. For more information, refer to Remington (The Science and Practice of Pharmacy, as above).

[0283] Gel formulations are semisolid suspension-type systems. Single-phase gels contain organic macromolecules substantially uniformly distributed throughout a carrier liquid, which is usually aqueous but preferably contains an alcohol and optionally an oil. Preferred organic macromolecules, i.e., gelling agents, are cross-linked acrylic acid polymers, such as the carbomer polymer family, such as those available under the trademark Carbopol. TM Other types of preferred polymers in this context are hydrophilic polymers such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers and polyvinyl alcohol; modified celluloses such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate and methylcellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. To prepare a uniform gel, a dispersant such as an alcohol or glycerol may be added, or the gelling agent may be dispersed by grinding, mechanical mixing or stirring, or a combination thereof.

[0284] Sprays typically provide the active agent in the form of an aqueous and / or alcoholic solution that can be atomized onto the skin for delivery. Such sprays include those formulated to provide a concentrated active agent solution at the site of administration after delivery, for example, the spray solution may consist primarily of alcohol or other similar volatile liquids in which the active agent may be dissolved. After delivery to the skin, the carrier evaporates, leaving the concentrated active agent at the site of administration.

[0285] Foam compositions are typically formulated as a single or multiphase liquid and packaged in a suitable container, optionally together with a propellant which aids in the expulsion of the composition from the container, thereby converting it to foam upon application. Other foam forming techniques include, for example, "bag-in-can" formulation techniques. Compositions so formulated typically contain a low boiling hydrocarbon, such as isopropane. Application and agitation of such compositions at body temperature causes the isopropanol to evaporate and produce foam in a manner similar to pressurized aerosol foaming systems. Foams can be water-based or aqueous alkanol, but are typically formulated with a high alcohol content which, when applied to the user's skin, evaporates rapidly, facilitating the active ingredient to pass through the upper layers of the skin to the treatment site.

[0286] Skin patches typically include a backing to which a reservoir containing an active agent is attached. The reservoir can be, for example, a pad that is dispersed or soaked with an active agent or composition, or a liquid reservoir. The patch also typically includes a front water-permeable adhesive that adheres and secures the device to the treatment area. Silicone rubber with self-adhesive properties can also be used. In both cases, a protective permeable layer can be used to protect the adhesive side of the patch before the patch is used. The skin patch can also include a removable cover to protect it during storage.

[0287] Examples of patch configurations that can be used for the present invention include single or multi-layer drugs in adhesive systems, characterized in that the drug is directly contained in the skin contact adhesive. In this transdermal patch design, the adhesive is not only used to fix the patch on the skin, but also used as a formulation basis, containing the drug and all excipients under a single backing film. In a multi-layer drug adhesive patch, the film is arranged between two different drug adhesive layers, or multiple drug adhesive layers are integrated under a single backing film.

[0288] Examples of pharmaceutically acceptable carriers suitable for topical pharmaceutical compositions include carrier materials well known in the cosmetic and medical fields as bases for, for example, emulsions, creams, aqueous solutions, oils, ointments, pastes, gels, lotions, milks, foams, suspensions, aerosols, etc., depending on the final form of the composition. Thus, representative examples of suitable carriers according to the present invention include, but are not limited to, water, liquid alcohols, liquid glycols, liquid polyalkylene glycols, liquid esters, liquid amides, liquid protein hydrolysates, liquid alkylated protein hydrolysates, liquid lanolin and lanolin derivatives, and similar materials commonly used in cosmetic and pharmaceutical compositions. Other suitable carriers according to the present invention include, but are not limited to, alcohols, such as monohydric and polyhydric alcohols, such as ethanol, isopropanol, glycerol, sorbitol, 2-methoxyethanol, diethylene glycol, ethylene glycol, hexylene glycol, mannitol and propylene glycol; ethers, such as diethyl ether or dipropyl ether; polyethylene glycol and methoxypolyoxyethylene (carbon wax with a molecular weight of 200 to 20,000); polyoxyethylene glycerol, polyoxyethylene sorbitol, stearyl diacetin, etc.

[0289] If desired, the topical compositions of the present disclosure may be contained in a package or dispenser device, such as a kit approved by the FDA, which may contain one or more unit dosage forms containing active ingredients. The dispenser device may, for example, include a tube. The package or dispenser device may be accompanied by instructions for administration. The package or dispenser device may also be accompanied by a notice specified by a government agency that regulates the manufacture, use or sale of drugs, which reflects the agency's approval of the form of the composition for human or veterinary administration. For example, such notices may include labels for prescription drugs approved by the U.S. Food and Drug Administration or approved product instructions. Compositions comprising the topical compositions of the present invention formulated in a pharmaceutically acceptable carrier may also be prepared, placed in an appropriate container, and labeled to treat a specified condition.

[0290] Another kind of patch system configuration that the present invention can use is a reservoir transdermal system design, it is characterized in that comprising a liquid compartment, wherein containing drug solution or suspension, separated from release liner by semipermeable membrane and adhesive.The adhesive component of this patch system can be added as the continuous layer between film and the peeling liner, and also can add with the concentric configuration around film.Another kind of patch system configuration that the present invention can use is a matrix system design, it is characterized in that comprising the semisolid matrix containing drug solution or suspension that directly contacts with release liner.The composition responsible for skin adhesion is integrated in the covering layer, and forms concentric configuration around the semisolid matrix.

[0291] Pharmaceutical compositions of the present disclosure can be in a form suitable for rectal administration, wherein the carrier is a solid. Preferably, the mixture forms a unit dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be conveniently formed by first mixing the composition with a softened or molten carrier, then cooling and molding in a mold.

[0292] Pharmaceutical compositions containing a compound of the present disclosure and / or a pharmaceutically acceptable salt thereof may also be prepared in the form of a powder or liquid concentrate.

[0293] Pharmaceutical compositions (or formulations) can be packaged in a variety of ways. Typically, articles for distribution include containers containing the pharmaceutical composition in an appropriate form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), pouches, foil blister packs, etc. The container may also include a tamper-proof component to prevent inadvertent access to the contents of the package. In addition, the container is typically labeled to describe the contents of the container and any appropriate warnings or instructions.

[0294] If desired, the disclosed pharmaceutical compositions may be contained in a package or dispenser device, which may contain one or more unit dosage forms containing the active ingredient. The package may, for example, include metal or plastic foil, such as a blister pack. The package or dispenser device may be accompanied by instructions for administration. The package or dispenser may also be accompanied by a notice associated with the container, the form of which is prescribed by a governmental agency that regulates the manufacture, use or sale of drugs, which reflects the agency's approval of the form of the drug for human or veterinary use. For example, such a notice may be a prescription drug label approved by the U.S. Food and Drug Administration, or it may be an approved product insert. Pharmaceutical compositions comprising a disclosed compound formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled to treat a specified condition.

[0295] The exact dosage and frequency of administration depends on the specific compound disclosed, the product of the disclosed preparation method, its pharmaceutically acceptable salt, solvate or polymorph, its hydrate, its solvate, its polymorph or its stereochemical isomeric form; the specific condition to be treated and the severity of the condition to be treated; various factors specific to the medical history of the subject to be administered, such as the age, weight, sex, degree of disease and general physical condition of the specific subject, and other drugs that the individual may be taking; this is well known to those skilled in the art. In addition, it is obvious that the effective daily dose can be lowered or increased according to the response of the subject to be treated and / or the evaluation of the physician prescribing the compound according to the present disclosure.

[0296] Depending on the mode of administration, the pharmaceutical composition will contain 0.05 to 99 wt%, preferably 0.1 to 70 wt%, more preferably 0.1 to 50 wt% of the active ingredient, and 1 to 99.95 wt%, preferably 30 to 99.9 wt%, more preferably 50 to 99.9 wt% of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.

[0297] In one aspect, suitable dosage levels are generally about 0.01 to 1000 mg of the compound described herein per kg of patient body weight per day, which can be administered in a single or multiple doses. In various aspects, the dosage level will be about 0.1 to about 500 mg / kg per day, about 0.1 to 250 mg / kg per day, or about 0.5 to 100 mg / kg per day. Suitable dosage levels can be about 0.01 to 1000 mg / kg per day, about 0.01 to 500 mg / kg per day, about 0.01-250 mg / kg per day, about 0.05-100 mg / kg per day, or about 0.1-50 mg / kg per day. Within this range, the dosage can be 0.05 to 0.5, 0.5 to 5.0, or 5.0 to 50 mg / kg per day. For oral administration, the composition is preferably provided in the form of tablets containing 1.0 to 1000 mg of active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000 mg of active ingredient, for adjusting the dosage of the patient to be treated according to the symptoms. The compound can be administered 1 to 4 times a day, preferably once or twice a day. This dosage regimen can be adjusted to provide the best therapeutic response.

[0298] The unit dose as described above and below can be administered more than once a day, for example 2, 3, 4, 5 or 6 times a day. In various aspects, such unit doses can be administered 1 or 2 times a day, so that for a 70 kg adult, the total dose per administration is in the range of 0.001 to about 15 mg / kg of subject body weight. On the other hand, the dose per administration is 0.01 to about 1.5 mg per kg of subject body weight, and such treatment can continue for weeks or months, and in some cases even for years. However, it is understood that the specific dose level for any particular patient will depend on various factors, including the activity of the specific compound used; the age, weight, general health, sex and diet of the individual being treated; the time and route of administration; the rate of excretion; other drugs previously administered; and the severity of the specific disease being treated, which is well known to those skilled in the art.

[0299] Typical dosages may be a single daily administration of a 1 mg to about 100 mg tablet or a 1 mg to about 300 mg tablet, or multiple daily administrations, or a single release capsule or tablet containing a higher proportion of the active ingredient per day. The time release effect may be obtained by capsule materials that dissolve at different pH values, capsules that release slowly by osmotic pressure, or by any other known controlled release means.

[0300] In some cases, it may be necessary to use dosages outside of these ranges, as will be apparent to one skilled in the art. In addition, it is noted that the clinician or treating physician will know how and when to initiate, interrupt, adjust or terminate treatment in conjunction with the response of an individual patient.

[0301] The disclosed pharmaceutical compositions may also contain other therapeutically active compounds, which are usually used to treat the above-mentioned pathological or clinical conditions.

[0302] It is to be understood that the disclosed compositions can be prepared from the disclosed compounds. It is also to be understood that the disclosed compositions can be used in the disclosed methods of use.

[0303] As mentioned above, the present invention relates to a pharmaceutical composition comprising an effective therapeutic amount of the disclosed compound, the product of the disclosed preparation method, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof and a pharmaceutically acceptable carrier. In addition, the present invention relates to a method for preparing such a pharmaceutical composition, characterized in that a pharmaceutically acceptable carrier is fully mixed with an effective therapeutic amount of the compound of the present invention.

[0304] The subject's cancer treatment method

[0305] In one aspect, disclosed herein is a method for treating a subject's cancer, the method comprising administering to the subject an effective therapeutic amount of at least one disclosed compound or a pharmaceutically acceptable salt thereof or a disclosed pharmaceutical composition. In some aspects, the subject is a human. In another aspect, the subject is diagnosed as needing to treat cancer before the administration step. In some aspects, the method further comprises the step of identifying a subject needing to treat cancer. In one aspect, the cancer is selected from non-small cell lung cancer (NSCLC), neuroblastoma, glioblastoma multiforme, metastatic brain cancer, brain cancer, prostate cancer or breast cancer.

[0306] In one aspect, the compounds described herein can treat brain metastases (BM). Brain metastases remain a significant problem in lung cancer. 6 The therapeutic efficacy of most small molecule targeted therapies is severely limited by blood-brain barrier (BBB) ​​efflux transporters. 6-10 Unfortunately, in NSCLC, up to 40% of patients develop brain metastases (BM), a number that is likely to increase as treatment options continue to improve life expectancy for patients with advanced disease. 6 Therefore, BM is a significant risk and contributes to poor prognosis in NSCLC patients treated with tyrosine kinase inhibitors (TKIs) with poor central nervous system (CNS) penetration. 7-10 As described herein, the compounds described herein exhibit high brain penetration and potent activity in osimertinib-resistant cell lines.

[0307] In another aspect, disclosed herein is a method of inhibiting epidermal growth factor receptor (EGFR) in a subject, comprising administering to the subject an effective therapeutic amount of at least one disclosed compound or a pharmaceutically acceptable salt thereof or a disclosed pharmaceutical composition. In one aspect, the subject is a human.

[0308] In one aspect, the compound is orally administered to a subject. In another aspect, the compound is administered at a dosage of about 50 mg / day to about 1000 mg / day, or about 50 mg / day, 50 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day and 750 mg / day, 800 mg / day, 850 mg / day, 900 mg / day, 950 mg / day or 1000 mg / day, wherein any value can be the lower and upper endpoints of a range (e.g., 100 mg / day to 300 mg / day).

[0309] As shown in the examples, the compounds described herein lack mutagenicity and show metabolic stability comparable to gefitinib. In addition, as shown in cell assays, these compounds have improved permeability and reduced discharge. The compounds described herein can be used alone or in combination with other chemotherapeutic agents and / or radiotherapy.

[0310] aspect

[0311] Aspect 1. A compound having structure I or a pharmaceutically acceptable salt thereof

[0312]

[0313] in

[0314] R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0315] n is an integer from 1 to 5, wherein each R 2 are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, halide or alkoxy;

[0316] m is an integer from 1 to 3, wherein each R 3 are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, halide or alkoxy;

[0317] o is an integer from 1 to 10;

[0318] X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0319] Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0320] R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0321] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f 、R7 g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0322] Each Z is independently hydrogen or deuterium.

[0323] Aspect 2. The compound according to aspect 1, wherein X is O.

[0324] Aspect 3. The compound according to aspect 1 or 2, wherein Y is O.

[0325] Aspect 4. The compound according to aspect 1 or 2, wherein Y is NR 5 , where R 5 It is a C1 to C5 alkyl group.

[0326] Aspect 5. The compound according to aspect 1 or 2, wherein Y is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group.

[0327] Aspect 6. A compound according to any one of aspects 1 to 5, wherein R 1 It's hydrogen.

[0328] Aspect 7. A compound according to any one of aspects 1 to 6, wherein R 3 It is an alkoxy group.

[0329] Aspect 8. A compound according to any one of aspects 1 to 6, wherein R 3 is an alkoxy group, and m is 1.

[0330] Aspect 9. The compound according to any one of aspects 1-8, wherein o is an integer from 1 to 5.

[0331] Aspect 10. A compound according to any one of aspects 1-9, wherein R 2 It is a halide, and n is 2.

[0332] Aspect 11. A compound according to any one of aspects 1 to 10, wherein R 2 It is an ortho fluoride.

[0333] Aspect 12. The compound according to Aspect 1, wherein the compound has structure II or a pharmaceutically acceptable salt thereof

[0334]

[0335] in

[0336] R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0337] R 2a and R 2b It is a halide;

[0338] R 3 is an alkoxy group;

[0339] o is an integer from 1 to 5;

[0340] X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0341] Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0342] R 6a and R 6bare independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0343] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0344] Each Z is independently hydrogen or deuterium.

[0345] Aspect 13. The compound according to aspect 12, wherein X is O.

[0346] Aspect 14. The compound according to aspect 12 or 13, wherein Y is O.

[0347] Aspect 15. The compound according to aspect 12 or 13, wherein Y is NR 5 , where R 5 It is a C1 to C5 alkyl group.

[0348] Aspect 16. The compound according to aspect 12 or 13, wherein Y is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group.

[0349] Aspect 17. A compound according to any one of aspects 12 to 16, wherein R 1 It's hydrogen.

[0350] Aspect 18. A compound according to any one of aspects 12 to 17, wherein R 3 is a C1 to C10 substituted or unsubstituted straight or branched chain alkoxy group.

[0351] Aspect 19. A compound according to any one of aspects 12 to 17, wherein R 3 It's a methoxy group.

[0352] Aspect 20. The compound according to any one of aspects 12 to 19, wherein o is an integer from 1 to 5.

[0353] Aspect 21. A compound according to any one of aspects 12 to 20, wherein R 2 It is a halide.

[0354] Aspect 22. A compound according to any one of aspects 12 to 21, wherein R 2a is chloride, R 2b It's fluoride.

[0355] Aspect 23. A compound according to any one of aspects 12 to 21, wherein R 2a is fluoride, R 2b It's chloride.

[0356] Aspect 24. The compound according to Aspect 1, wherein the compound has structure III or a pharmaceutically acceptable salt thereof

[0357]

[0358] in

[0359] R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0360] R 2a and R 2b It is a halide;

[0361] R 3 is an alkoxy group;

[0362] o is an integer from 1 to 5;

[0363] X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0364] Y is O, NR 5 or CR 6a R6 b , where R 5 is hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0365] R 6a and R 6b are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0366] R 7a , R 7b , R 7c , R 7d , R 7e , R7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0367] Each Z is independently hydrogen or deuterium.

[0368] Aspect 25. The compound according to aspect 24, wherein X is O.

[0369] Aspect 26. The compound according to aspect 24 or 25, wherein Y is O.

[0370] Aspect 27. The compound according to aspect 24 or 25, wherein Y is NR 5 , where R 5 It is a C1 to C5 alkyl group.

[0371] Aspect 28. The compound according to aspect 24 or 25, wherein Y is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group.

[0372] Aspect 29. A compound according to any one of aspects 24-18, wherein R 1 It's hydrogen.

[0373] Aspect 30. A compound according to any one of aspects 24 to 29, wherein R 3 is a C1 to C10 substituted or unsubstituted straight or branched chain alkoxy group.

[0374] Aspect 31. A compound according to any one of aspects 24 to 29, wherein R 3 It's a methoxy group.

[0375] Aspect 32. The compound according to any one of aspects 24-31, wherein o is an integer from 1 to 5.

[0376] Aspect 33. A compound according to any one of aspects 24-32, wherein R 2 It is a halide.

[0377] Aspect 34. A compound according to any one of aspects 24 to 32, wherein R 2a is chloride, R 2b It's fluoride.

[0378] Aspect 35. A compound according to any one of aspects 24 to 32, wherein R 2a is fluoride, R 2b It's chloride.

[0379] Aspect 36. The compound according to aspect 1, wherein the compound is

[0380]

[0381] Aspect 37. A pharmaceutical composition comprising a compound according to any one of claims 1 to 36 and a pharmaceutically acceptable carrier.

[0382] Aspect 38. A method for treating a subject having non-small cell lung cancer, neuroblastoma, glioblastoma multiforme, metastatic brain cancer, brain cancer, breast cancer or prostate cancer, the method comprising administering to the subject an effective amount of a compound according to any one of Aspects 1 to 36.

[0383] Aspect 39. A method of inhibiting epidermal growth factor receptor (EGFR) in a subject, the method comprising administering to the subject an effective amount of a compound according to any one of aspects 1 to 36.

[0384] Aspect 40. A method of treating a subject having osimertinib-resistant cancer, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 36.

[0385] Aspect 41. A method of treating a subject having brain metastatic cancer, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 36.

[0386] Aspect 42. The method according to any one of aspects 38 to 41, wherein the compound is administered orally to the subject.

[0387] Aspect 43. The method according to any one of aspects 38 to 42, wherein the compound is administered at a dose of about 50 mg per day to about 1000 mg per day.

[0388] Aspect 44. A method for preparing the compound described in any one of aspects 1 to 16, the method comprising reacting a compound having structure IV with a compound having structure V in the presence of a base.

[0389]

[0390] in

[0391] R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0392] R 2a and R 2b It is a halide;

[0393] R 3 is an alkoxy group;

[0394] o is an integer from 1 to 5;

[0395] X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl;

[0396] Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and

[0397] R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino;

[0398] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl;

[0399] each Z is independently hydrogen or deuterium; and

[0400] LG is a leaving group.

[0401] Aspect 45. The method according to aspect 44, wherein LG is a halogen or a sulfonic acid group.

[0402] Aspect 46. The method according to aspect 44 or 45, wherein the base comprises a hydride, an alkoxide, a Grignard reagent, or an alkyl lithium compound.

[0403] Aspect 47. A method for preparing a compound having structure X

[0404]

[0405] in

[0406] R 5 is a substituted or unsubstituted straight chain or branched chain alkyl group, and

[0407] R7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0408] each Z is independently hydrogen or deuterium,

[0409] The method comprises

[0410] (a) reacting a compound having structure XI with a compound having structure XII in the presence of a base to produce a compound having structure XIII

[0411]

[0412] Where R 10 is a substituted or unsubstituted straight chain or branched chain alkyl group, or a substituted or unsubstituted straight chain or branched chain alkoxy group, and

[0413] LG is a leaving group;

[0414] (b) reacting a compound having structure XIII with a first oxidant under heating to produce a compound having structure XIV

[0415]

[0416] (c) reacting a compound having structure XIV with (i) a second oxidizing agent and then with (ii) a first reducing agent to produce a first intermediate; and

[0417] (d) reacting the first intermediate with a second reducing agent to produce a compound having structure x.

[0418] Aspect 48. The method according to aspect 47, wherein R 10 It is a C1-C5 straight chain or branched chain alkyl group.

[0419] Aspect 49. The method according to aspect 47, wherein R 10 It is a C1-C5 straight chain or branched chain alkoxy group.

[0420] Aspect 50. The method according to aspect 47, wherein R 10 It is a C1-C5 straight or branched chain alkoxy group substituted by an aryl group.

[0421] Aspect 51. The method according to aspect 47, wherein R 10 It is benzyloxy.

[0422] Aspect 52. The method according to any one of aspects 46 to 51, wherein LG is a halide, a sulfonate, a carbonate or a phosphate.

[0423] Aspect 53. The method according to any one of aspects 47-52, wherein the base in step (a) comprises a carbonate, a hydroxide, a phosphate, a hydride, a dialkylamide or hexamethyldisilazane.

[0424] Aspect 54. The method according to any one of aspects 47 to 53, wherein step (a) is performed in an aprotic organic solvent.

[0425] Aspect 55. The method according to any one of aspects 47-54, wherein step (a) is performed at a temperature of about 25°C to about 100°C.

[0426] Aspect 56. The method according to any one of aspects 47-55, wherein step (b) comprises the following steps:

[0427] (i) reacting a compound having structure XIII with a first oxidizing agent in a first organic solvent to prepare a first composition;

[0428] (ii) adding an aqueous base to the first composition to prepare a second composition comprising an organic layer and an aqueous layer;

[0429] (iii) separating the organic layer from the aqueous layer;

[0430] (iv) removing the first organic solvent from the organic layer to produce a residue;

[0431] (v) dissolving the residue in a second organic solvent to prepare a second composition; and

[0432] (vi) heating the second composition from about 50°C to about 100°C to produce a compound having structure XIV.

[0433] Aspect 57. The method according to aspect 56, wherein the first organic solvent is dichloromethane and the second organic solvent is toluene.

[0434] Aspect 58. The method according to any one of aspects 47 to 57, wherein the first oxidizing agent in step (b) comprises a peroxyacid, ozone, or hydrogen peroxide / acetic acid.

[0435] Aspect 59. The method according to any one of aspects 47 to 57, wherein the first oxidizing agent in step (b) comprises meta-chloroperbenzoic acid.

[0436] Aspect 60. The method according to any one of aspects 47-57, wherein the molar ratio of the first oxidizing agent to the compound having structure XIII is from 0.95:1 to 1:1.05.

[0437] Aspect 61. The method according to any one of aspects 47-60, wherein step (c) comprises the following steps:

[0438] (i) reacting a compound having structure XIV with a second oxidizing agent in a third organic solvent to produce a third composition; and

[0439] (ii) mixing the first reducing agent with the third composition to prepare a first intermediate.

[0440] Aspect 62. The method according to aspect 61, wherein the second oxidant comprises ozone or osmium tetroxide and sodium periodate.

[0441] Aspect 63. The method according to aspect 61 or 62, wherein the third organic solvent comprises an alcohol and an aprotic solvent.

[0442] Aspect 64. The method according to any one of aspects 61-63, wherein the compound having structure XIV is reacted with the second oxidant at a temperature of about -50°C to about -100°C.

[0443] Aspect 65. The method according to any one of aspects 47-64, wherein the first reducing agent comprises a hydride.

[0444] Aspect 66. The method according to any one of aspects 47-64, wherein the first reducing agent comprises a borohydride.

[0445] Aspect 67. The method according to any one of aspects 47-66, wherein the molar ratio of the first reducing agent to the compound having structure XIV is from 1.5:1 to 2.5:1.

[0446] Aspect 68. The method according to any one of aspects 47 to 67, wherein the first intermediate is isolated before step (d).

[0447] Aspect 69. The method according to any one of aspects 47-68, wherein step (d) comprises the following steps:

[0448] (i) dissolving the first intermediate in an aprotic solvent to prepare a fourth composition;

[0449] (ii) mixing a second reducing agent with the fourth composition to produce a compound having structure X; and

[0450] (iii) isolating and purifying the compound having structure X.

[0451] Aspect 70. The method of aspect 69, wherein the second reducing agent is mixed with the fourth composition at a temperature of about 10°C to about -50°C.

[0452] Aspect 71. The method according to any one of aspects 47-70, wherein the second reducing agent comprises a hydride.

[0453] Aspect 72. The method according to any one of aspects 47-70, wherein the second reducing agent comprises aluminum hydride.

[0454] Aspect 73. The method according to any one of aspects 47-70, wherein the molar ratio of the second reducing agent to the first intermediate is 4:1 to 2:1.

[0455] Aspect 74. The method according to any one of aspects 47-73, wherein R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are hydrogen respectively.

[0456] Aspect 75. The method of any one of Aspects 47-73, wherein R 5 is methyl, R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h They are hydrogen.

[0457] Aspect 76. A method for preparing a compound having structure XX

[0458]

[0459] in

[0460] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0461] each Z is independently hydrogen or deuterium,

[0462] The method comprises

[0463] (a) reacting a compound having structure XXI with a compound having structure XXII in the presence of a base to prepare a compound having structure XXIII

[0464]

[0465] Where LG is the leaving group;

[0466] (b) reacting a compound having structure XXIII with a first oxidant to produce a compound having structure XXIV

[0467] and

[0468] (c) reacting a compound having structure XXIV with (i) a second oxidizing agent and then with (ii) a first reducing agent to produce a compound having structure XX.

[0469] Aspect 77. The method according to aspect 76, wherein LG is a halide, a sulfonate, a carbonate or a phosphate.

[0470] Aspect 78. The method according to aspect 76 or 77, wherein the base in step (a) comprises a carbonate, a hydroxide, a phosphate, a hydride, a dialkylamide or hexamethyldisilazane.

[0471] Aspect 79. The method according to any one of aspects 76 to 78, wherein step (a) is performed in an aprotic organic solvent.

[0472] Aspect 80. The method according to any one of aspects 76-79, wherein step (a) is performed at a temperature of about 25°C to about 100°C.

[0473] Aspect 81. The method according to any one of aspects 76 to 80, wherein step (b) comprises the following steps:

[0474] (i) reacting a compound having structure XXIII with a first oxidizing agent in a first organic solvent to prepare a first composition;

[0475] (ii) adding an aqueous base to the first composition to prepare a second composition comprising an organic layer and an aqueous layer;

[0476] (iii) separating the organic layer from the aqueous layer;

[0477] (iv) removing the first organic solvent from the organic layer to produce a residue;

[0478] (v) dissolving the residue in a second organic solvent to prepare a second composition; and

[0479] (vi) heating the second composition from about 50°C to about 100°C to produce a compound having structure XXIV.

[0480] Aspect 82. The method according to aspect 81, wherein the first organic solvent is dichloromethane and the second organic solvent is toluene.

[0481] Aspect 83. The method according to any one of aspects 76 to 82, wherein the first oxidizing agent in step (b) comprises a peroxyacid, ozone, or hydrogen peroxide / acetic acid.

[0482] Aspect 84. The method according to any one of aspects 76 to 83, wherein the first oxidizing agent in step (b) comprises meta-chloroperbenzoic acid.

[0483] Aspect 85. The method according to any one of aspects 76-83, wherein the molar ratio of the first oxidizing agent to the compound having structure XIII is from 0.95:1 to 1:1.05.

[0484] Aspect 86. The method according to any one of aspects 76-84, wherein step (c) comprises the following steps:

[0485] (i) reacting a compound having structure XXIV with a second oxidizing agent in a third organic solvent to produce a third composition; and

[0486] (ii) mixing the first reducing agent with the third composition to produce a compound having structure XX; and

[0487] (iii) isolating and purifying the compound having structure XX.

[0488] Aspect 87. The method according to aspect 86, wherein the second oxidant comprises ozone or osmium tetroxide and sodium periodate.

[0489] Aspect 88. The method according to aspect 86 or 87, wherein the third organic solvent comprises an alcohol and an aprotic solvent.

[0490] Aspect 89. The method according to any one of aspects 86-88, wherein the compound having structure XXIV is reacted with the second oxidant at a temperature of about -50°C to about -100°C.

[0491] Aspect 90. The method according to any one of aspects 76-89, wherein the first reducing agent comprises a hydride.

[0492] Aspect 91. The method according to any one of aspects 76-89, wherein the first reducing agent comprises a borohydride.

[0493] Aspect 92. The method according to any one of aspects 76-91, wherein the molar ratio of the first reducing agent to the compound having structure XXIV is from 1.5:1 to 2.5:1.

[0494] Aspect 93. The method according to any one of aspects 76-92, wherein R 7a , R 7b , R 7c , R 7d , R 7e , R 7f 、R7 g and R 7h are hydrogen respectively.

[0495] Aspect 94. The method according to any one of aspects 47-73, wherein R 5 is methyl, R 7a , R 7b , R 7c , R 7d , R 7e , R 7f 、R7 g and R 7h are hydrogen respectively.

[0496] Aspect 95. Compounds having structure XXX

[0497]

[0498] in

[0499] Y is O or NR 5 ,

[0500] R 5 is a substituted or unsubstituted straight chain or branched chain alkyl group, and

[0501] R 7a , R 7b , R 7c , R 7d , R 7e , R 7f 、R7 g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and

[0502] Each Z is independently hydrogen or deuterium.

[0503] Having now described many aspects of the present disclosure, in general, the following examples describe some additional aspects of the present invention. Although aspects of the present disclosure are described in conjunction with the following examples and corresponding text and drawings, it is not intended that the various aspects of the present disclosure be limited to this description. On the contrary, this application is intended to cover all alternatives, modifications and equivalents included within the spirit and scope of the present disclosure.

[0504] Example

[0505] The following examples are presented to provide a person of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are prepared and evaluated, and are intended to be purely exemplary of the present disclosure and are not intended to limit the scope of the disclosure that the inventors consider to be disclosed. Efforts have been made to ensure the accuracy of numbers (such as quantities, temperatures, etc.), but some errors and deviations should be considered. Unless otherwise stated, parts are by weight, temperatures are in degrees Celsius or at ambient temperature and pressure.

[0506] Example 1

[0507] General Experimental Unless otherwise stated, all reactions were performed in oven-dried glassware covered with rubber septa under argon atmosphere. All organic solutions were concentrated under reduced pressure on a rotary evaporator and water bath. Flash column chromatography was performed using silica gel (Fisher silica gel adsorbent (230-400 mesh, 60 grade)). Thin layer chromatography (TLC) was performed using 250 μM glass-backed silicon (XHL) plates with a fluorescent indicator (254 nm). TLC plates were visualized by exposure to ultraviolet light (UV) and / or immersion in cerium ammonium molybdate (CAM) in ethanol followed by heating on a hot plate (120° C., 10-15 s).

[0508] The starting material 2-(tert-butyldimethylsilyloxy)-ethan-1-ol was purchased from AK Scientific. 11-132-Hydroperoxy-2-methyltetrahydro-2H-pyran (MTHPOOH) was prepared and the spectral data were consistent with those previously reported in the literature. Morpholine and N-methylpiperazine were purchased from Sigma-Aldrich. Isopropylmagnesium chloride-lithium chloride complex solution (1.3 M in THF) was purchased from Sigma-Aldrich. Ethylmagnesium bromide (3 M in THF) was purchased from Sigma-Aldrich. Tetrabutylammonium fluoride (1 M in THF) was purchased from TCI. Thionyl chloride was purchased from AlfaAesar. Sodium hydride (60% dispersion in mineral oil) was purchased from Sigma-Aldrich. 4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ol was purchased from AK Scientific. 4-(3-Chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-ol was purchased from AmBeed. Gefitinib (1) (≥98% (HPLC), SML1657-50MG) was purchased from Sigma-Aldrich. The materials used for in vitro analysis are given in the "In vitro Analytical Methods" section of the Examples section. The purity of the tested compounds (6, 13-15) was >95% as determined by UHPLC and NMR.

[0509] Instrument Nuclear magnetic resonance (NMR) spectra of all compounds were obtained at 25°C in CDCl3 (δ 7.26 and 77.16 ppm, respectively) or DMSO-D6 (δ 2.50 and 39.52 ppm, respectively) using a 500 MHz, EZC500 JEOL instrument. Chemical shifts (δ) are calculated relative to the residual solvent peak and are expressed in ppm. Multiplicity abbreviations are as follows: s (singlet), m (multiplet), b (broad), d (doublet), t (triplet), q (quartet), hept (septet). High-resolution mass spectra were obtained on a ThermoFisher Orbitrap Q-Exactive using electrospray ionization (ESI). Melting points were determined on a Barstead Electrothermal 9100. ThermoFisher Vanquish UHPLC with a PDA detector and Acclaim 120 18 UHPLC traces of 8, 13-15 were obtained using a C 4.6 x 50 mm column and the percent purity was determined using the Avalon peak area algorithm. The instrumentation used in the in vitro analysis is given in the In Vitro Analytical Methods section of the Examples section.

[0510] Through literature method 14 Synthesis of tert-butyldimethyl(2-(2-methyltetrahydro-2H-pyran-2-yl)peroxy)ethoxy)silane (8)

[0511]

[0512] At 0 ℃, pyridine (9.66mL, 113.57mmol, 1.5 equivalents) was added to the stirred solution of anhydrous DCM (250mL) containing 7 (13.33g, 75.71mmol, 1.0 equivalents), then Tf was added dropwise O (15.26mL, 90.85mmol, 1.2 equivalents) and the solution was stirred at 0 ℃ for 45 minutes. After this time, the mixture was diluted with DCM (100mL), and washed with 1NHCl (1x, 150mL) aqueous solution successively. NaHCO (1x, 150mL) and saline solution (1x, 150mL). The organic layer combined was dried with Na SO, filtered and concentrated in vacuo. The crude triflate was directly used in the next step without further purification.

[0513] At 0°C, add MTHPOOH 11-13 To a stirred solution of KO (12 g, 90.85 mmol, 1.2 eq.) in anhydrous THF (260 mL) was added t Bu (10.19 g, 90.85 mmol, 1.2 eq.), the solution was stirred for 30 minutes, and then the crude triflate obtained in the previous step was added dropwise to anhydrous THF (40 mL). The solution was heated to room temperature and stirred for 90 minutes. After this, the mixture was diluted with EtOAc (100 mL) and quenched by adding NaHCO3 (1x, 200 mL). The layers were separated and the aqueous layer was extracted with EtOAc (2x, 100 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The obtained residue was purified by flash column chromatography on silica gel (eluent: EtOAc: hexane at 10:90) to give the title compound 8 (11.19 g, 38.56 mmol, 51%) as a yellow oil. The spectral data were consistent with those previously reported in the literature. 14 .

[0514] R f =0.50 (20:80 ethyl acetate:hexane; CAM).

[0515] 1 H NMR (500MHz, CDCl3): δ4.13-4.07(m,2H),3.92(td,J=11.4,2.8Hz,1H),3.85(t,J=5.4 Hz,2H),3.71-3.68(m,1H),1.79-1.50(m,6H),1.43(s,3H),0.90(s,9H),0.08(s,6H).

[0516] 13C NMR (126MHz, CDCl3): δ102.5,76.7,61.8,60.8,33.3,26.0,24.9,24.6,19.2,18.5,-5.2.

[0517] Synthesis of 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)morpholine (10)

[0518]

[0519] To a stirred solution of morpholine (7.64 mL, 87.33 mmol, 3.0 equiv) in anhydrous THF (73 mL) was added i PrMgCl LiCl (1.3M in THF) (56mL, 72.78mmol, 2.5 equivalents), the solution was allowed to reach room temperature and stirred for 45 minutes. After this time, 8 (8.45g, 29.11mmol, 1.0 equivalents) was added dropwise to anhydrous THF (73mL) and the solution was stirred for 3 hours. After this time, the solution was quenched with NaHCO (100mL). The layers were separated and the aqueous layer was extracted with EtOAc (3x, 75mL). The combined organic layers were dried with NaSO, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel (eluent: EtOAc: hexane at 15:85) to give the title compound 10 (4.95g, 18.95mmol, 65%) as a pale yellow oil.

[0520] R f =0.20 (10:90 EtOAc:hexanes; CAM).

[0521] 1 H NMR (500MHz, CDCl3): δ3.88 (d, J = 11.8Hz, 2H), 3.79-3.72 (m, 4H), 3.62-3.54 (m, 2H) ), 3.15 (d, J = 10.6Hz, 2H), 2.66 (td, J = 10.9, 3.3Hz, 2H), 0.89 (s, 9H), 0.06 (s, 6H).

[0522] 13 C NMR (126MHz, CDCl3): δ73.1, 66.4, 61.6, 56.4, 26.0, 18.5, -5.1.

[0523] HRMS-ESI(m / z):[C 12 H 28 O3N 28 Si] + [M+H] +Calculated value: 262.1833, measured value: 262.1835.

[0524] 1-(2-((tert-Butyldimethylsilyl)oxy)ethoxy)-4-methylpiperazine (16)

[0525]

[0526] To a stirred solution of N-methylpiperazine (12.62 mL, 113.73 mmol, 3.0 equiv) in anhydrous THF (95 mL) was added i PrMgCl·LiCl (1.3M THF solution) (72.9mL, 94.78mmol, 2.5 equivalents), the solution was adjusted to room temperature and stirred for 45 minutes. After this time, 8 (11g, 37.91mmol, 1.0 equivalents) was added dropwise to anhydrous THF (95mL) and the solution was stirred for 3 hours. After this time, the solution was quenched with NaHCO3 (75mL). The layers were separated and the aqueous layer was extracted with EtOAc (3x, 50mL). The combined organic layers were dried with Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified by silica gel flash column chromatography (eluent: 40:55:5 hexane: EtOAc: Et3N) to give the title compound 16 (6.99g, 25.5mmol, 67%) as a yellow oil.

[0527] R f =0.30 (40:55:5 hexane:EtOAc:Et3N; CAM).

[0528] 1 H NMR(500MHz,C6D6)δ3.83(t,J=5.2Hz,2H),3.75(t,J=5.5Hz,2H),3.15-3.13(m,2H),2.8 4(brs,2H),2.49-2.46(m,2H),2.10-2.06(m,2H),2.02(s,3H),0.99(s,9H),0.09(s,6H).

[0529] 13 C NMR (126MHz, C6D6) δ73.2, 62.0, 55.9, 54.4, 45.6, 26.2, 18.6, -5.1.

[0530] HRMS-ESI(m / z):[C 13 H 31 O2N2Si] + [M+H] + Calculated value: 275.2149, measured value: 275.2146.

[0531] Synthesis of 2-(Morpholinooxy)ethan-1-ol (11)

[0532]

[0533] To a stirred solution of anhydrous THF (150 mL) containing 10 (4.90 g, 18.80 mmol, 1.0 eq) was added TBAF (1 M in THF) (37.60 mL, 37.60 mmol, 2.0 eq) dropwise at 0° C. The solution was brought to room temperature and stirred for 1 hour. After this time, the mixture was concentrated in vacuo and the resulting residue was purified by flash column chromatography (eluent: 95:5 EtOAc:Et3N) to give the title compound 11 (2.17 g, 14.75 mmol, 79%) as a yellow oil.

[0534] R f =0.50(95:5EtOAc:Et3N;CAM),

[0535] 1 H NMR (500MHz, CDCl3): δ3.91 (d, J=12.3Hz, 2H), 3.86-3.79 (m, 4H), 3.56 (t, J=12.6Hz, 2H), 3.34 (br s, 1H), 3.21 (d, J = 11.9Hz, 2H), 2.67 (td, J = 10.9, 3.3Hz, 2H).

[0536] 13 C NMR (126MHz, CDCl3): δ71.6, 66.3, 63.6, 56.2.

[0537] HRMS-ESI(m / z):[C6H 14 O3N] + [M+H] + Calculated value: 148.0968, measured value: 148.0964.

[0538] 2-((4-Methylpiperazin-1-yl)oxy)ethan-1-ol (17)

[0539]

[0540] To a stirred solution of anhydrous THF (300 mL) containing 16 (6.35 g, 23.16 mmol, 1.0 eq) was added TBAF (1 M in THF) (46.32 mL, 46.32 mmol, 2.0 eq) dropwise at 0° C. The solution was brought to room temperature and stirred for 90 min. After this time, the mixture was concentrated in vacuo and the resulting residue was purified by flash column chromatography (eluent: 90:10 EtOAc:Et3N) to give the title compound 17 (2.18 g, 13.61 mmol, 71%) as a yellow oil.

[0541] R f =0.10(90:10EtOAc:Et3N;CAM).

[0542] 1 H NMR (500MHz, toluene-D8) δ3.69-3.66(m,2H),3.65-3.60(m,2H),3.01(br d,J=10.3Hz,2H),2.73-2.59(m,2H),2.41(br d,J=11.3Hz,2H),1.96(s,3H),1.94-1.90(m,2H).

[0543] 13 C NMR (126 MHz, toluene-D8) δ 72.2, 63.2, 55.4, 54.2, 45.3.

[0544] HRMS-ESI(m / z):[C7H 17 O2N2] + [M+H] + Calculated value: 161.1285, measured value: 161.1282.

[0545] Synthesis of N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(2-(morpholinooxy)ethoxy)quinazolin-4-amine (6)

[0546]

[0547] To a stirred solution of 11 (1 g, 6.80 mmol, 1.0 eq) in anhydrous toluene (20 mL) was added dropwise SOCl (1.23 mL, 17.00 mmol, 2.5 eq) at 0° C., stirred until exothermic, then warmed to 60° C. and stirred for 3 hours. After this time, the mixture was concentrated in vacuo and the residue was used directly in the next step without further purification.

[0548] To a stirred solution of 4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ol (3.26 g, 10.20 mmol, 1.5 eq) in anhydrous DMF (35 mL) at 0°C, NaH (60% dispersion in mineral oil) (406 mg, 10.20 mmol / L, 1.5 eq) was slowly added and the solution was warmed to room temperature and stirred for 45 minutes. Afterwards, the crude chloride obtained in the previous step was added dropwise to DMF (5 mL), the solution was raised to 80°C and stirred for 16 hours. After this, the mixture was concentrated in vacuo and co-concentrated with toluene (3x, 50 mL). The resulting residue was then redissolved in EtOAc (200 mL), washed with 1 M NaOH (2x, 100 mL), then brine (2x, 100 ml), and the combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The obtained residue was purified by flash column chromatography (eluent: 10:90 Et3N:EtOAc) to give the title compound 6 (1.08 g, 2.41 mmol, 35% after two steps) as a light yellow powder.

[0549] R f =0.50(10:90Et3N:EtOAc; UV,CAM).

[0550] 1 H NMR (500MHz, DMSO-D6): δ9.51(s,1H),8.50(s,1H),8.12(dd,J=6.8,2.7Hz,1H),7.83-7.78(m,2H),7.44(t,J=9.1Hz,1H),7.20(s,1H),4.29(t, J=4.5Hz,2H),4.08(t,J=4.5Hz,2H),3.94(s,3H),3.81(d,J=11.6Hz,2H),3.44(t,J=11.4Hz,2H)3.18(d,J=10.4Hz,2H),2.54(d,J=11.8Hz,2H).

[0551] 13 C NMR (126MHz, DMSO-D6): δ156.0,154.5,153.1(d, 1 J C-F =243.2Hz),152.7,148.2,147.0,136.8(d, 3 J C-F =3.8Hz),123.4,122.2(d, 3 J C-F =6.3Hz),118.8(d, 2 J C-F=18.9fHz),116.5(d, 2 J C-F =21.4Hz),108.7,107.4,102.7,69.0,67.4,65.5,56.2,55.8.

[0552] 13 C NMR 19 F} (126MHz, DMSO-D6): δ156.0,154.5,153.1,152.7,148.2,147.0,136.8,12 3.4,122.2,118.8,116.5,108.7,107.4,102.7,69.0,67.4,65.5,56.2,55.8.

[0553] 19 F{ 1 H}(470MHz,DMSO-D6):-123.2.

[0554] HRMS-ESI(m / z):[C 21 H 23 O4N4 35 ClF] + [M+H] + Calculated value: 449.1386, measured value: 449.1378.

[0555] Melting point: 186.4-187.5°C (n=average value of 3 measurements)

[0556] Synthesis of N-(3-chloro-2-fluorophenyl)-7-methoxy-6-(2-(morpholinooxy)ethoxy)quinazolin-4-amine (13)

[0557]

[0558] To a stirred solution of 11 (1.90 g, 12.9 mmol, 1.0 eq) in anhydrous toluene (40 mL) was added dropwise SOCl (3.84 mL, 32.25 mmol, 2.5 eq) at 0° C., stirred until exothermic, then warmed to 60° C. and stirred for 3 hours. After this time, the mixture was concentrated in vacuo and the residue was used directly in the next step without further purification.

[0559] To a stirred solution of anhydrous DMF (70 mL) containing 4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-ol (6.19 g, 19.35 mmol, 1.5 eq) was slowly added NaH (60% dispersion in mineral oil) (770 mg, 19.35 mmol / L, 1.5 eq) at 0°C, the solution was heated to room temperature and stirred for 45 minutes. Afterwards, the crude chloride obtained in the previous step was added dropwise to DMF (10 mL), the solution was raised to 80°C and stirred for 16 hours. After this, the mixture was concentrated in vacuo and co-concentrated with toluene (3x, 75 mL). The resulting residue was then redissolved in EtOAc (150 mL), washed with 1 M NaOH (2x, 75 mL), brine (2x, 75 mL), and the combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The obtained residue was purified by flash column chromatography (eluent: 10:90 Et3N:EtOAc) to give the title compound 13 as an off-white powder (1.67 g, 3.73 mmol, 29% after two steps).

[0560] R f =0.50(10:90Et3N:EtOAc; UV,CAM).

[0561] 1 H NMR (500MHz, DMSO-D6): δ9.61(s,1H),8.38(s,1H),7.82(s,1H),7.54-7.52(m,1H),7.49-7.46(m,1H),7.28(t,J=8.3Hz,1H),7.21(s,1H),4.28 (t,J=4.6Hz,2H),4.07(t,J=4.6Hz,2H),3.94(s,3H),3.81(d,J=12.0Hz,2H),3.44(t,J=11.3Hz,2H),3.17(d,J=10.4Hz,2H),2.55-2.52(m,2H).

[0562] 13 C NMR (126MHz, DMSO-D6): δ156.9,154.6,153.0,152.4(d, 1 J C-F =249.5Hz),148.2,147.0,128.4(d, 2 J C-F =12.6Hz),127.1,126.9,124.9(d, 3 J C-F =5.0Hz),120.2(d, 2 JC-F =16.4Hz),108.6,107.2,102.8,69.0,67.3,65.5,56.2,55.9.

[0563] 13 C NMR 19 F}(126MHz,DMSO-D6): δ156.9,154.6,153.0,152.4,148.2,147.0,128.4,12 7.1,126.9,124.9,120.2,108.6,107.2,102.8,69.0,67.3,65.5,56.2,55.9.

[0564] 19 F{ 1 H}(470MHz,DMSO-D6):--120.4.

[0565] HRMS-ESI(m / z):[C 21 H 23 O4N4 35 ClF] + [M+H] + Calculated value: 449.1386, measured value: 449.1376.

[0566] Synthesis of N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(2-(4-methylpiperazin-1-yl)oxy)ethoxy)quinazolin-4-amine (14)

[0567]

[0568] At 0 ° C, SOCl2 (340 L, 4.68 mmol, 2.5 eq.) was added dropwise to a stirred solution of anhydrous toluene (7 mL) containing 17 (300 mg, 1.87 mmol, 1.0 eq.), stirred until no longer exothermic, then raised to 60 ° C and stirred for 3 hours. After this time, the mixture was concentrated in vacuo, and the resulting residue was dissolved in EtOAc (20 mL) and washed with K2CO3 aqueous solution (2x, 15 mL). The combined organic layer was then dried over Na2SO4, filtered and concentrated. The resulting residue was used without further purification.

[0569] To a stirred solution of anhydrous DMF (10 mL) containing 4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ol (898 mg, 2.81 mmol, 1.5 eq) was slowly added NaH (60% dispersion in mineral oil) (112 mg, 2.81 mmol, 1.5 eq) at 0°C, the solution was heated to room temperature and stirred for 45 minutes. Afterwards, the crude chloride obtained in the previous step was added dropwise to DMF (5 mL), the solution was raised to 80°C and stirred for 16 hours. After this, the mixture was concentrated in vacuo and co-concentrated with toluene (3x, 20 mL). The resulting residue was then redissolved in EtOAc (30 mL), washed with 1 M NaOH (2x, 20 mL), brine (2x, 20 mL), and the combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography (eluent: 85:10:5 EtOAc:MeOH:Et3N) to afford the title compound 14 as a light orange powder (286 mg, 0.687 mmol, 33% after two steps).

[0570] R f =0.40 (85:10:5EtOAc:MeOH:Et3N; UV, CAM).

[0571] 1 H NMR (500MHz, DMSO-D6): δ9.53(s,1H),8.50(s,1H),8.12(dd,J=6.8,2.6Hz,1H),7.84-7.78(m,2H),7.44(t,J=9.1Hz,1H ),7.21(s,1H),4.28(t,J=5.0Hz,2H),4.04(t,J=5.0Hz,2H),3.94(s,3H),3.15-3.13(m,2H),2.70-2.67(m,2H),2.57(br s,2H),2.13-2.09(m,5H).

[0572] 13 C NMR (126MHz, DMSO-D6): δ156.0,154.5,153.1(d, 1 J C-F =243.2Hz),152.7,148.2,147.0,136.8,126,123.4,122.2(d, 3 J C-F =6.3Hz),118.8(d, 2 J C-F =18.9Hz),116.5(d, 2 J C-F=21.4Hz),108.7,107.4,102.7,69.1,67.4,55.9,55.0,53.5,45.0.

[0573] 13 C NMR 19 F} (126MHz, DMSO-D6): δ156.0,154.5,153.1,152.7,148.2,147.0,136.8,123.4 ,122.2,118.8,116.5,108.7,107.4,102.7,69.0,67.4,55.9,55.0,53.5,45.0.

[0574] 19 F{ 1 H}(470MHz,DMSO-D6):-123.2.

[0575] HRMS-ESI(m / z):[C 22 H 26 O3N5 35 ClF] + [M+H] + Calculated value: 462.1703, measured value: 462.1691.

[0576] Melting point: 64.0-66.5°C (n=average value of 3 measurements)

[0577] Synthesis of N-(3-chloro-2-fluorophenyl)-7-methoxy-6-(2-(4-methylpiperazin-1-yl)oxy)ethoxy)quinazolin-4-amine (15)

[0578]

[0579] At 0 ° C, SOCl2 (2.38, mL, 32.80 mmol, 2.5 eq.) was added dropwise to a stirred solution of anhydrous toluene (50 mL) containing 17 (2.10 g, 13.12 mmol, 1.0 eq.), stirred until no longer exothermic, then raised to 60 ° C and stirred for 3 hours. After this time, the mixture was concentrated in vacuo, the resulting residue was dissolved in EtOAc (100 mL) and washed with K2CO3 aqueous solution (2x, 75 mL). The combined organic layer was then dried with Na2SO4, filtered and concentrated. The resulting residue was used without further purification.

[0580] To a stirred solution of anhydrous DMF (90 mL) containing 4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-ol (6.29 g, 19.68 mmol, 1.5 eq) was slowly added NaH (60% dispersion in mineral oil) (783 mg, 19.68 mmol / L, 1.5 eq) at 0°C, the solution was warmed to room temperature and stirred for 45 minutes. Afterwards, the crude chloride obtained in the previous step was added dropwise to DMF (10 mL), the solution was warmed to 80°C and stirred for 16 hours. After this, the mixture was concentrated in vacuo and co-concentrated with toluene (3x, 25 mL). The resulting residue was then redissolved in EtOAc (150 mL) and washed with 1M NaOH (2x, 100 mL), brine solution (2x, 100 mL), and the combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The obtained residue was purified by flash column chromatography (eluent: 85:10:5 EtOAc:Et3N:MeOH) to give the title compound 15 (1.19 g, 2.58 mmol, 20% after two steps) as a light yellow powder.

[0581] R f =0.30 (85:10:5EtOAc:MeOH:Et3N; UV, CAM).

[0582] 1 H NMR (500MHz, DMSO-D6): δ9.61(s,1H),8.39(s,1H),7.83(s,1H),7.57-7.51(m,1H),7.48-7.46(m,1H),7.29-7.26(m,1H ),7.21(s,1H),4.27(t,J=4.6Hz,2H),4.03(t,J=4.6Hz,2H),3.94(s,3H),3.15-3.12(m,2H),2.69-2.67(m,2H),2.57(br s,2H),2.13-2.06(m,5H).

[0583] 13 C NMR (126MHz, DMSO-D6): δ156.9,154.6,153.0,152.4(d, 1 J C-F =249.5Hz),148.2,147.0,128.4(d, 2 J C-F =11.3Hz),127.1,126.8,124.9(d, 3 J C-F =5.0Hz),120.2(d, 2 JC-F =16.4Hz),108.6,107.2,102.8,69.0,67.2,55.9,55.0,53.6,45.0.

[0584] 13 C NMR 19 F}(126MHz,DMSO-D6): δ156.9,154.6,153.0,152.4,148.2,147.0,128.4,127.1 ,126.8,124.9,120.1,108.6,107.2,102.8,69.0,67.2,55.9,55.0,53.6,45.0.

[0585] 19 F{ 1 H}(470MHz,DMSO-D6):-120.4.

[0586] HRMS-ESI(m / z):[C 22 H 26 O3N5 35 ClF] + [M+H] + Calculated value: 462.1703, measured value: 462.1695.

[0587] Melting point: 143.5-144.9°C (n=average value of 3 measurements)

[0588] Alternative Synthesis of Piperazinyl Hydroxylamine

[0589] Figure 3 An exemplary synthetic scheme for preparing the piperazinyl analogs described herein is provided in. [2,3]-Meisenheimer rearrangement for installation of the hydroxylamine unit 15-17 Starting from commercially available 18, allylation with allyl bromide under basic conditions gave the N-allyl derivative (19) in 72% yield. N-oxidation of 19 with m-chloroperbenzoic acid (MCPBA) followed by [2,3]-Meisenheimer rearrangement afforded 20 in 47% yield after 2 steps. The olefin present in 20 was then ozonolyzed and reduced with sodium borohydride (NaBH4) followed by lithium aluminum hydride. 18 Further reduction of the carboxybenzyl group by (LiAlH4) afforded the desired hydroxylamine precursor 17 in 50% yield after 2 steps.

[0590] experiment

[0591] Synthesis of 4-allylpiperazine-1-carboxylic acid benzyl ester (19)

[0592]

[0593] To a stirred solution of anhydrous THF (250 mL) containing piperazine-1-carboxylic acid benzyl ester (15 g, 68.14 mmol, 1.0 eq) was added K2CO3 (18.8 g, 136.28 mmol, 2.0 eq) at room temperature, followed by the slow addition of allyl bromide (11.8 mL, 136.28 mmol, 2.0 eq). The solution was raised to 65 ° C and stirred for 16 hours, after which the mixture was cooled to room temperature and quenched by the addition of 1M NaOH (200 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3x, 100 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel (eluent: 50:45:5 hexane: EtOAc: Et3N) to give the title compound 19 (12.81 g, 49.2 mmol, 72%) as a yellow oil. TLC R f =0.60 (50:45:5 hexane:EtOAc:Et3N; CAM, UV). 1 H NMR (500MHz, CDCl3) δ7.37-7.29(m,5H),5.89-5.78(m,1H),5.23-5.14(m,2H),5.13(br s,2H),3.56(m,4H),3.00(d,J=6.6Hz,2H),2.40(br s,4H). 13 C NMR (126MHz, CDCl3) δ155.4,136.9,134.7,128.6,128.1,128.0,118.5,67.2,61.8,52.8,43.9.HRMS-ESI(m / z): [C 15 H 21 N2O2] + [M+H] + Calculated value: 261.1598, measured value: 261.1594.

[0594] Synthesis of Benzyl 4-(Allyloxy)piperazine-1-carboxylate (20)

[0595]

[0596] At -30 ℃, add MCPBA (mixture of 3-chlorobenzoic acid and water containing 70% MCPBA) (11.56g, 46.9mmol, 1.0 equivalent) in the stirring solution of anhydrous DCM containing 19 (12.2g, 1.0 equivalent) in batches, continue 5 minutes.After adding completely, the reaction mixture was stirred for 1 hour at -30 ° ℃, then quenched with NaHCO (150mL) aqueous solution, separate layers. Wash the organic layer with NaHCO (3x, 150mL), then use Na SO Dry the organic layer, filter and vacuum concentrate. The residue obtained is dissolved in the toluene heated to 80 ℃ and stirred for 14 hours. After this time, the mixture is cooled to room temperature and vacuum concentrated. The resulting residue was purified by flash column chromatography on silica gel (eluent: 80:15:5 hexanes:EtOAc:Et3N) to afford the title compound 20 (6.13 g, 22.2 mmol, 47%) as a clear oil. TLC R f =0.50 (30:70 EtOAc:hexane; CAM, UV). 1 H NMR(500MHz, CDCl3)δ7.39-7.30(m,5H)5.99-5.88(m,1H),5.27(d,J=17.3Hz,1H),5.18(d,J= 10.3Hz,1H),5.13(s,2H),4.21(d,J=6.1Hz,2H),4.03(s,2H),3.19-3.11(m,4H),2.57(s,3H). 13 C NMR (126MHz, CDCl3): δ155.3,136.8,134.6,128.7,128.2,128.1,118.0,73.2,67.4,55.3,42.7.HRMS-ESI(m / z):[C 15 H 21 N2O3] + [M+H] + Calculated value: 277.1547, measured value: 277.1540.

[0597] Synthesis of 2-((4-methylpiperazin-1-yl)oxy)ethan-1-ol (17)

[0598]

[0599] Compound 20 (5.6 g, 20.27 mmol, 1.0 equiv) was dissolved in a mixture of DCM (200 mL) and methanol (50 mL) and cooled to -78 ° C. Ozone was bubbled in the reaction mixture for 30 minutes until the starting material was completely consumed. After completion, as shown in ESIMS, argon was bubbled in the reaction mixture to disperse the residual ozone. NaBH4 (1.53 g, 40.54 mmol, 2.0 equiv) was then added in batches at -78 ° C, and the solution was gradually heated to room temperature and stirred for 1 hour. After this time, the mixture was quenched with NaHCO3 (200 mL) aqueous solution and the layers were separated. The aqueous layer was extracted with DCM (2x, 150 mL), and the combined organic layers were washed with brine (2x, 150 mL) aqueous solution, dried with Na2SO4, filtered and concentrated in vacuo to directly obtain a clear oily alcohol (5.26 g, 18.8 mmol, 93% crude yield). TLC R f =0.10 (70:30 EtOAc:hexane; CAM, UV). 1 H NMR (500MHz, CDCl3) δ7.38-7.30(m,5H),5.13(s,2H),4.07(s,2H),3.84-3.81(m,4H),3.24-3.10(m,5H),2.60-2.56(m,2H). 13 C NMR (126MHz, CDCl3) δ155.2,136.6,128.7,128.3,128.1,71.9,67.5,65.3,55.0,42.7.HRMS-ESI(m / z): [C 14 H 21 O4N2] + [M+H] + Calculated value: 281.1496, measured value: 281.1489.

[0600] The crude alcohol (5.26 g, 18.8 mmol, 1.0 equiv) obtained in the previous step was dissolved in anhydrous THF (68 mL) and cooled to 0 ° C. LiAlH4 (2.31 g, 60.81 mmol, 3.2 equiv) was then added in batches over 5 minutes. After complete addition, the solution was heated to room temperature and stirred for 2 hours. After this time, the reaction mixture was cooled to 0 ° C and quenched by slowly adding H2O. After effervescence stopped, the solution was acidified to pH ≈ 2 with concentrated HCl. The aqueous layer was then extracted with EtOAc (4x, 150 mL), after which the aqueous layer was cooled to 0 ° C and alkalized to pH ≈ 10 by adding solid NaOH in batches. The basified aqueous layer was extracted with EtOAc (4x, 100 mL), and the resulting organic layer was dried over Na2SO4, filtered and concentrated in vacuo to afford the title compound 17 (1.63 g, 10.2 mmol, 50% yield in two steps starting from compound 20) as a yellow oil, whose spectral data were identical to those of the authentic sample.

[0601] In vitro analytical methods

[0602] Kinase activity

[0603] Through Eurofins DiscoverX (K d Assay) and Eurofins Cerep (Biochemical IC 50 In vitro kinase activity (inhibitor binding constant (K)) was assessed by d ) and biochemical inhibition (IC 50 )). For K d Compounds were assayed in duplicate (n=2) using Eurofin's KINOMEScan KdELECT assay using an 11-point, 3-fold dilution series at the highest compound tested concentration of 10 μM. 50 Assays, compounds were run in duplicate (n=2) and tested in an enzyme radiometric assay using a 9-point, half-log dilution series with the highest compound tested at 10 μM and an ATP concentration of 10 μM using Eurofin's KinaseProfiler technology.

[0604] LogD 7.4 (Shake flask method) (Pharmaron Co., Ltd., Ningbo, China)

[0605] Compound lipophilicity was determined by Pharmaron Inc. using a shake flask method with 1-octanol and PBS (pH 7.4). Each compound was evaluated in duplicate (n=2) at 1 μM in 96-well plates with shaking at 2000 rpm for 2 hours at 25°C. Samples were analyzed by LC-MS / MS.

[0606] Solubility determination in PBS at pH 7.4 (Pharmaron Co., Ltd., Ningbo, China)

[0607] Solubility was determined by Pharmaron Inc. Each compound was evaluated in duplicate (n=2) by adding 15 μL of compound stock solution (10 mM in DMSO) to a 96-well plate with 485 μL of buffer and shaking at 25°C, 1100 rpm for 2 hours. The wells were filtered, samples (5 μL) were taken, and then diluted with an equal volume of DMSO (5 μL) and 490 μL of aqueous solution. Samples were analyzed by LC-MS / MS.

[0608] Metabolic stability analysis (Pharmaron Co., Ltd., Ningbo, China)

[0609] Pharmaron Co., Ltd. used a compound depletion method to evaluate the metabolic stability of compounds in incubations containing human and rat liver microsomes and hepatocytes. Compound 15 was additionally evaluated for stability in monkey and dog hepatocytes. Microsomal stability was evaluated twice (n=2) using liver microsomes (0.5 mg / mL) from humans (BD Gentest) and SD rats (BD Gentst) in the presence and absence of NADPH. The compound was incubated in liver microsomes at 1 μM and samples (30 μL) were taken at time points of 0.5, 15, 30, 45 and 60 minutes and analyzed by LC-MS / MS. At 0.5×10 6 Hepatocyte stability was assessed in duplicate (n=2) using hepatocytes from humans (BiolVT), SD rats (BiolVT), cynomolgus monkeys (RILD), or beagle dogs (BiolVT) at a working cell density of 10 cells / mL. Compounds were incubated in hepatocytes at 1 μM and samples (25 μL) were taken at 0.5, 15, 30, 60, 90, and 120 minutes and analyzed by LC-MS / MS. In vitro half-life and intrinsic clearance were determined as previously described 19 .

[0610] In vitro half-life (in vitro t 1 / 2 ) is determined by the slope value: in vitro t 1 / 2 =-(0.693 / k).

[0611] where k is determined by linear regression of the natural logarithm of the percentage of parent drug remaining versus the incubation time curve.

[0612] Plasma protein binding determined by equilibrium dialysis (Pharmaron Co., Ltd., Ningbo, China)

[0613] Plasma protein binding and brain tissue binding were determined by Pharmaron Co., Ltd. using equilibrium dialysis. Each compound was evaluated twice at 5 μM (n=2), and the final volume percentage of organic solvent was 0.5%. The compound was incubated at 37°C, 5% CO2 and 100 rpm for 6 hours, and samples (50 μL) were taken at the beginning and end of the incubation. After incubation, the samples were analyzed by LC-MS / MS, and the concentration of the compound was determined in the buffer and plasma solution chambers.

[0614] Plasma stability (Pharmaron Co., Ltd., Ningbo, China)

[0615] The compounds were evaluated by Pharmaron Co., Ltd. using a compound depletion method to determine the stability of the compounds in incubations containing human (Pharmaron) and SD rat (IPHASE) plasma. The compounds were evaluated twice (n=2) and incubated in 5 μM plasma, sampled (50 μL) at 0, 15, 30, 60 and 120 minutes, and analyzed by LC-MS / MS. The peak area ratio was determined from the extracted ion chromatogram, and the percentage of the compound remaining at each time point was calculated by the following equation.

[0616] Remaining Percent t min (%) = Peak area ratio t min / Peak Area Ratio 0min ×100

[0617] Among them, the peak area ratio t min is the peak area ratio of the control and the test compound at t min, and peak area ratio 0min is the peak area ratio of the control and the test compound at time point zero.

[0618] Permeability studies (Pharmaron Co., Ltd., Ningbo, China)

[0619] The ability of compounds to penetrate Caco-2 cells and MDCKII-MDR1 cells was evaluated at Pharmaron Inc. 96-well HTS Transwell plates (Corning) were used for seeding Caco-2 and MDCKII-MDR1 cells. Caco-2 cells were plated at 6.86 × 10 5 Cells were seeded at a density of 1.56×10 cells / mL and cultured for 14-18 days before testing. 6MDCKII-MDR1 cells were inoculated at a density of 10 cells / mL and cultured for 4-8 days before detection. In order to determine the transport rate of the drug in the absorption (apical to basolateral (AB)) and secretion (basolateral to apical (BA)) directions, the compound (DMSO containing 5 μM Caco-2 cells, DMSO containing 1 μM MDCKII-MDR1 cells) was added to the donor well. The plate was incubated at 37°C for 2 hours, and samples (50 μL) were taken in the donor and receptor wells at the beginning and end of the incubation, and two measurements (n=2) were performed. The samples were analyzed by LC-MS / MS. The apparent permeability coefficient (P) was calculated using the following equation: app ), unit is cm / s×10 -6 .

[0620] P app =V A ×[Drug] 受体 / area × time × [drug] 初始、供体

[0621] Among them, V A = represents the volume in the receptor well (mL). Area is the surface of the membrane (0.143 cm for a Transwell 96-well plate). 2 ), time is the total transit time (seconds).

[0622] The efflux ratio was determined using the following equation: efflux ratio = P app (BA) / P app (AB)

[0623] Among them, P app (BA) represents the apparent permeability from basolateral to apical direction, while P app (AB) represents the apparent permeability from apical to basolateral direction.

[0624] hERG channel inhibition assay

[0625] The inhibitory potential of the compounds on the hERG channel was evaluated by Pharmaron GmbH using a manual patch clamp system as described previously. 20 HEK293 cell line stably transfected with hERG gene (Invitrogen) was used. Compounds were tested at 5 concentrations (0.37, 1.11, 3.33, 10 and 30 μM) and repeated three times (n=3). IC 50 IC values ​​were determined by plotting compound concentration versus inhibition rate (%) using GraphPad Prism software and fitting a nonlinear regression equation to the sigmoidal dose-response curve. 50 Values ​​are expressed as mean ± SEM.

[0626] Determination of human CYP450 inhibition using compound 15

[0627] Pharmaron GmbH evaluated the potential of compound 15 to inhibit cytochrome P450 (CYP) isoforms using human liver microsomes. The activities tested were CYP1A2-mediated O-demethylation of phenacetin, CYP2C19-mediated 4'-hydroxylation of (S)-mephenytoin, CYP2C9-mediated 4'-hydroxylation of diclofenac, CYP2D6-mediated O-demethylation of dextromethorphan, and CYP3A4-mediated 1'-hydroxylation of midazolam. Substrate concentrations were phenacetin (40 μM), mephenytoin (50 μM), diclofenac (6 μM) and dextromethorphan (2 μM), and midazolam (1 μM). The probe substrates phenacetin, mephenytoin, and dextromethorphan were incubated at 37°C for 20 minutes. The probe substrates diclofenac and midazolam were incubated at 37°C for 5 minutes. Compound (15) was tested in an eight-point, three-fold dilution series (0.01 μM to 30 μM) in DMSO and incubated with pooled human liver microsomes (0.5 mg / mL) (BD Gentest) and a selective CYP isomer probe substrate mix. After a 5-minute preincubation, the reaction was initiated by the addition of NADPH (final concentration of 1 mM) and assayed in duplicate (n=2). Samples were analyzed by UPLC-MS / MS. Inhibition of each P450 enzyme was measured as the percent reduction in labeled metabolite formation activity compared to an uninhibited control. IC was determined on GraphPad Prism. 50 The values, logarithms of residual activity (%) and inhibitor concentration were fitted using a nonlinear fit [inhibitor], using a normalized response model with a variable slope.

[0628] Compound 15 to determine the time-dependent inhibition of CYP2D6

[0629] Compound 15 was evaluated using human liver microsomes (BD Gentest) and primary human hepatocytes (BiolVT) to determine potential time-dependent inhibition of CYP2D6. Bufurol (2 μM) was used as a substrate for liver microsomes and dextromethorphan (40 μM) for hepatocytes. The activities tested were CYP2D6-mediated 1'-hydroxylation of bufurol and CYP2D6-mediated O-demethylation of dextromethorphan. Compound 15 was assayed twice (n=2) and tested in a six-point, three-fold dilution series (0.03 μM to 10 μM) in DMSO and mixed with either human liver microsomes (0.5 mg / mL) or human hepatocytes (0.3×10 6Cells / mL) were incubated together with pooled human liver microsomes. Pooled human liver microsomes were incubated with NADPH (final concentration 1 mM) at 37°C, 5% CO2 with or without a 30 min preincubation. Incubations in human hepatocytes were performed without preincubation or at 37°C, 5% CO2 for 30 min. The experiments were performed at 37°C, 5% CO2 for 5 min. Inhibition of CYP2D6 was measured as the percent reduction in activity of labeled metabolite formation compared to an uninhibited control. IC was determined on GraphPadPrism 50 The values, logarithms of residual activity (%) and inhibitor concentration were fitted using a nonlinear fit [inhibitor], using a normalized response model with a variable slope.

[0630] AMES Volatility Test (Eurofins Panlabs, St. Charles, Missouri, USA)

[0631] The mutagenicity of the compounds was assessed at Eurofins Panlabs. Four Salmonella strains were used in 384-well plates as described previously: TA98, for probing frameshift mutations (with quercetin as control), TA100 and TA1535, for probing base pair insertions / deletions (with streptozotocin as control), and TA1537, for probing frameshift mutations (with aminoacridine as control). 21 Each compound was incubated at 37°C for 96 h at four different concentrations (5, 10, 50 and 100 μM), with 48 replicates per concentration tested with or without rat liver S9 metabolic activation. Bacterial cytotoxicity assays were performed simultaneously at 0.6, 1.2, 2.5, 5, 10, 25, 50 and 100 μM to exclude false negatives. Bacterial cytotoxicity was expressed as a percentage of control growth (OD 650 Compounds showing growth less than 60% of the control were marked and considered toxic to bacterial cells. Wells showing bacterial growth due to reversal of the histidine mutation (based on OD 430 / OD 570 The ratio of 1 to 2 was greater than 1.0 (judgment) and recorded as positive counts. The significance of positive counts between the treatment group and the control group was determined using a one-tailed Fisher's exact test.

[0632] In vitro micronucleus assay

[0633] Eurofins Panlabs evaluated the genotoxicity of compound 15. The in vitro micronucleus assay using Chinese hamster ovary (CHO-K1) cells was performed as described previously. 22Compound 15 was incubated at different concentrations (8, 16, 31, 62, 125, 250, 500 and 1000 μM) in 96-well plates at 37°C for 4 h (rat liver S9 for metabolic activation) and at 37°C for 24 h (rat liver S9 without metabolic activation). High content analysis and fluorescence imaging were used to detect micronuclei and compared with positive controls (cyclophosphamide (performed at 7.2 μM) and mitomycin C (performed at 0.3 μM)). The significance of positive counts between treatment and control groups was determined by a two-sample one-tailed t-test with equal variance.

[0634] Kinase selectivity assay compound 15

[0635] A 468-member human kinase panel was analyzed using the KINOMEScan platform at Eurofins DiscoverX. Compound 15 was tested against the 468 kinases at a single concentration of 1 μM. TM Percent control was mapped onto the kinome tree. S scores were calculated as previously described. 23 , the number of kinases that bind to 15 out of the total number of wild-type kinases.

[0636] In vitro CTG assay (Crown Bioscience)

[0637] Cell lines

[0638] A431, HCC827, SK-BR-3, ZR-75-30, AU565 and Caco-2 cell lines were purchased from ATCC. NCI-H1975 cell line was purchased from SIBS. NCI-H3255 cell line was purchased from CoBioer. Engineered Ba / F3 cell line was purchased from Crown Bioscience. MCKII-MDR1 cells were obtained from the Netherlands Cancer Institute. HEK293 cells were purchased from Invitrogen. A431 cells were cultured in DMEM (Life Technologies) containing 10% FBS. HCC827, NCI-H1975 and AU565 cells were cultured in RPMI1640 (Invitrogen) containing 10% FBS. NCI-H3255 cells were cultured in BEGM (Lonza) containing 10% FBS. Ba / F3 EGFR-del E746_A750 / C797S and Ba / F3 EGFR-L858R / C797S cells were cultured in RPMI (Invitrogen) containing 10% FBS. SK-BR-3 cells were cultured in McCoy's5a (Invitrogen) containing 10% FBS. ZR-75-30 cells were cultured in RPMI1640 (Invitrogen) containing 20% ​​FBS. BT474 cells were cultured in DMEM (Gibco) containing 10% FBS and 10 μg / mL insulin. The culture information of Caco-2 and MCKII-MDR1 can be found in the permeability study section. HEK293 cells were cultured in DMEM (Gibco) containing 10% FBS, 0.1 mM NEAA, 25 mM HEPES, 100 U / mL penicillin-streptomycin, μg / mL blasticidin, and 400 μg / mL geneticin. All cells were cultured in a humidified incubator at 37°C and 5% CO2.

[0639] Viability assays were performed at Crown BioScience using A431, HCC827, NCI-H1975, NCI-H3255, Ba / F3-EGFR delE746_A750 / C797S, Ba / F3 EGFR-L858R / C797S, AU565, SK-BR-3, and ZR-75-30 cells. Cells were seeded into 96-well plates at 1500-7000 cells per well and dosed with DMSO containing compounds (0.15 nM to 10 μM) in three doses (n=3) in a nine-point, four-fold dilution series and incubated for 72 hours. After 72 hours, cell viability was determined by the CellTiter-Glo luminescent activity assay (Promega). Dose response curves were generated and used to calculate IC 50 Value, IC50 Values ​​were calculated on GraphPad Prism from a nonlinear regression equation fitted with a sigmoidal dose response and are expressed as mean ± SEM.

[0640] Animal studies

[0641] Animal experiments were performed at Pharmaron Co., Ltd., and the Institutional Animal Care and Use Committee (IACUC) of Pharmaron approved the use of animals according to the guidance of AAALAC (Pharmaron IACUC, protocols #PK-M-07182022, #PK-R-06012022, and #ON-CEL-XEN-06012022). In pharmacokinetic studies, we used 6-8 week-old male SD rats (about 200-300 g) from Si Bei Fu Laboratory Animal Technology Co., Ltd., 6-8 week-old male CD1 mice (about 20-30 g) from Si Bei Fu Laboratory Animal Technology Co., Ltd., and 6-8 week-old female BALB / c nude mice (about 20-3 g) from Si Bei Fu Laboratory Animal Technology Co., Ltd. In the intracranial PDX study, 6-8 week-old female BALB / c nude mice (about 18-22 g) obtained from Beijing Anikeeper Biotechnology Co., Ltd. were used. The animals were housed at a temperature of 20-25°C and relative humidity between 40-70%, exposed to a 12-h light and dark cycle, and provided with food and water ad libitum.

[0642] In vivo studies

[0643] Pharmacokinetic study of compound 15

[0644] The standard pharmacokinetic evaluation of compound 15 was performed by IV (intravenous injection) tail vein injection (formulation: DMSO: saline solution containing 10% sulfobutyl-β-cyclodextrin = 1:99) and oral PO (oral) gavage (formulation: 1% methylcellulose), followed by blood sampling at 8 time points for IV injection (0.0833, 0.25, 0.5, 1, 2, 4, 7, 24 hours after administration) and 7 time points for oral administration (0.25, 0.5, 1, 2, 4, 8, 24 hours after administration), n = 3 animals for each route of administration (n = 6 in total). At each time point, approximately 0.03 mL of blood was collected from the dorsal plantar vein (for CD1 mice), 0.03 mL of blood was collected from the orbital vein (for BALB / c nude mice), or 0.2 mL of blood was collected from the jugular vein (for SD rats). Blood from each sampling point was transferred to a plastic microcentrifuge tube containing K2-EDTA. The collection tube containing the blood sample and anticoagulant was inverted several times to properly mix the contents of the tube and placed on ice before centrifuging the plasma. The blood samples were centrifuged at 4000g for 5 minutes at 4°C to obtain plasma. The samples were stored in a refrigerator at -75°C before analysis. The concentration of the test article in the plasma samples was determined by LC-MS / MS and analyzed using WinNonlin 8.3 (Phoenix TM ) were used for pharmacokinetic calculations. The values ​​obtained were plotted on GraphPad Prism software and expressed as mean ± SD.

[0645] The oral central nervous system pharmacokinetics of compound 15 were evaluated by oral gavage (formulation: 1% methylcellulose), followed by blood and brain sampling at 7 time points (0.25, 0.5, 1, 2, 4, 8, 24 hours after administration), with n = 3 animals at each time point (n = 21 in total). For plasma collection, approximately 0.03 mL of blood was collected from the plantar dorsal vein (for CD1 mice) or 0.2 mL of blood was collected from the jugular vein (for SD rats) at each time point. The blood at each sampling point was transferred to a plastic microcentrifuge tube containing K2-EDTA, and the collection tube containing the blood sample and anticoagulant was inverted several times to properly mix the contents of the tube and placed on ice before centrifuging the plasma. The blood samples were then centrifuged at 4°C, 4000g for 5 minutes to obtain plasma, and the samples were stored in a refrigerator at -75°C before analysis. For brain sample collection, the rodents were fully bled before collection, the chest cavity was opened, and the heart cannulated from the left ventricle was exposed. A small incision was then made in the right atrial appendage, and saline was gently injected via syringe. Brain samples were collected at each time point and quickly frozen in an ice box. Samples were stored at -75°C before analysis. All brain samples were prepared with water before analysis to achieve a ratio of brain weight (g) to water volume (mL) of 1:4. The concentration of 15 in plasma and brain samples was determined using LC-MS / MS and analyzed using WinNonlin 8.3 (Phoenix TM ) were used for pharmacokinetic calculations. The values ​​obtained were plotted on GraphPad Prism software and expressed as mean ± SD.

[0646] Intracranial PDX model of compound 15

[0647] HCC827 cells stably expressing luciferase (HCC827-luc) were injected intracranially. Briefly, 3×10 5HCC827-luc tumor cells were injected into the right forebrain of anesthetized mice (anesthetic: intramuscular injection of ZolettTM 50 (Virbac SA)). Mice were imaged once every two weeks using IVIS Lumina III (Perkin Elmer). Images were collected 10 minutes after intraperitoneal injection of 15 mg / mL (5 μL / g body weight) of D-luciferin in anesthetized mice (anesthetic: inhaled 1-2% isoflurane). On day 20 after cell inoculation, mice were randomly divided into 2 treatment groups (n=10 mice per group): 1) 1% methylcellulose as vehicle control, 2) 15 (10 mg / kg) (formulation: 1% methylcellulose), oral, twice daily (bid) administration (12 hours apart), 7 days, for 21 days. Mice will be euthanized in the following cases: 1) if a single animal shows obvious signs of severe distress and / or pain, 2) if body weight (BW) loss exceeds 20%, or 3) if the animal cannot obtain adequate food or water. The body weights of all mice were measured every two weeks throughout the study, and the body weight change (BW change) (expressed in %) was calculated using the following formula.

[0648] Body weight change (%) = (BW 第x天 / BW 第0天 )100

[0649] Among them, BW 第x天 is the body weight on a given date, BW 第0天 is the body weight on day 0 (start of treatment).

[0650] Statistical analysis

[0651] Statistical analysis was performed using GraphPadPrism 9.0. Data are presented as mean ± SD or SEM when n = ≥ 3, or as mean when n = 2. For in vitro ADME and kinase activity studies, data are presented as mean of n = 2 independent replicates. For in vivo PK experiments, data are presented as mean ± SD, with n ≥ 3 animals per study group. For in vitro short-term growth delay experiments, IC 50 Values ​​were determined by nonlinear regression equations fitted with sigmoidal dose-response curves and are expressed as mean ± SEM, n = 3 independent replicates. Two-tailed unpaired t-test was used to compare intracranial PDX study data (mean values ​​of bioluminescence and body weight at day 21 after treatment initiation, n = 10 animals per study group). p values ​​< 0.05 were considered statistically significant.

[0652] Results and discussion

[0653] like Figure 4 As shown, hydrooxazole (6) was prepared using our direct NO bond formation reaction.11 To this end, 2-methyltetrahydropyranyl (MTHP) monoperoxyacetal, derived in 51% yield from commercially available 2-((tert-butyldimethylsilyl)oxy)ethanol (7), was used as the catalyst for the reaction. 11-13 (8) was exposed to a morpholine-derivatized magnesium amide to afford the hydroxylamine (10) in 65% yield on an 8 g scale. 24 Developed turbo-Grignard ( i The use of a morpholine-derived magnesium amide (PrMgCl·LiCl) proved to be the best choice for large-scale formation of the N-O bond. While the generation of the morpholine-derived magnesium amide from EtMgBr provided the product in reproducible yields on a small scale, on a larger scale (>5 g), this magnesium amide showed solubility issues, thus prompting us to investigate alternatives (see the Examples section for more details). However, desilylation of 10 afforded the alcohol (11) in 79% yield, which was converted to the chloride by reaction with thionyl chloride and subsequently displaced with commercially available 4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ol under basic conditions to afford the expected hydroxalog (6) in 35% yield after 2 steps. Overall, the hydroxalog (6) was prepared from readily available starting materials in only the 4 longest linear steps (LLS), highlighted by the direct formation of the N-O bond, with all synthetic transformations being performed on a gram scale.

[0654] With 6 in hand, we proceeded to evaluate inhibitor binding constants and biochemical inhibition of the relevant EGFR kinase forms of 1 and 6, a direct hydroxylamine analog; both 1 and 6 exhibited single-digit nanomolar activity for activation of mutants bearing EGFR, indicating that the N-(normethoxy)morpholine unit is an effective bioisostere of the N-alkylmorpholine unit. We then evaluated in vitro ADME properties, starting with a permeability assay in colon cancer (Caco-2) cells, which express both P-gp and BCRP; 6 showed a 14-fold increase in permeability and a greatly reduced efflux compared to 1 (Table 1). In a permeability assay in MDCK II-MDR1 cells, 6 showed an approximately 4-fold increase in permeability and a reduced efflux compared to 1, an effect often used to mimic the blood-brain barrier by overexpressing the active efflux transporters P-gp or MDR1. These results suggest that the substitution of N-alkylmorpholine units with N-(normethoxy)morpholine units is a promising approach. 25 This may overcome the known poor BBB permeability and active efflux observed in 1. Of note, currently osimertinib (5), a third-generation EGFR TKI, is the only EGFR TKI approved for the treatment of EGFR+ NSCLC BM, despite being a substrate for both P-gp and BCRP. 8We then performed AMES fluctuation assays in four Salmonella strains (TA98, TA100, TA1537, and TA1535) with and without rat liver S9 metabolic activation (+ / - S9) and found that neither 1 or 6 were mutagenic. 26 These findings, combined with similar stability profiles of multiple species in liver microsomes and plasma, dispelled any early concerns regarding the potential mutagenicity and instability of the trisubstituted hydroxylamine unit in 6 with respect to 1.

[0655] We sought to further minimize efflux and improve drug-like properties, particularly the aqueous solubility of 6, by synthesizing additional trisubstituted hydroxylamine-containing inhibitors (see below). Figure 7-8 Given the importance of hydrogen bond donors (HBDs) for substrate recognition by efflux transporters and noting the improved CNS penetration of the EGFR inhibitors AZD3759 and JCN037, compound 13 was designed with a para- to ortho-fluorine switch to minimize the HBD capacity of the adjacent aniline ( Figure 7 ) 27-30 In addition, 14 and 15 were prepared by exchanging the morpholine unit with an N-methylpiperazine group while maintaining the key trisubstituted hydroxylamine moiety, with the goal of improving aqueous solubility and potential in vivo exposure. All of the newly prepared analogs maintained single-digit to subnanomolar binding and biochemical inhibition of activation of mutants bearing EGFR. In the Caco-2 cell assay, 13 and 15 bearing ortho-fluorine exhibited significantly enhanced permeability and reduced efflux ratios compared to 1, and even improved over 6 (Table 1). Notably, 14, which contains a more basic nitrogen heterocycle than 1, 6, and 13, with a para-fluorine substitution pattern on the aniline ring, showed high efflux and low permeability, highlighting the synergistic improvement of substrate recognition by efflux transporters with improved HBD and reduced pKa. In the MDCKII-MDR1 cell assay, 13 and 15 showed excellent permeability and low efflux compared to 1, 14, and even 6. Importantly, 15 also showed good stability in human and rat microsomes and hepatocytes, while its water solubility was significantly improved compared with 1 and 6.

[0656] We analyzed the anticancer activity of inhibitors with hydroxylamine against four patient-derived cell lines with different EGFR status, always with cisplatin as a positive control ( Fig. 9 ). It is noteworthy that 15 pairs of patients carrying common EGFR L858R The mutant NCI-H3255 NSCLC cell line showed excellent activity, IC 50 The value is 7.2nM, which is about 12 times higher than 6. delE746_A750 The potent activity of 15 was also observed in HCC827 NSCLC cells, with an IC50 is 3.1nM. L858R / C797S and EGFR delE746_A750 / C797S In the osimertinib-resistant Ba / F3 cell line engineered with C797S point mutation to block covalent inhibitor binding, 15 also showed strong activity, with IC 50 The values ​​were 4.6 and 2.5 nM respectively 31 Overall, 15 was a potent inhibitor in osimertinib-resistant engineered cell lines and patient-derived NSCLC cell lines NCI-H3255 and HCC827, which carry EGFR mutations and account for approximately 85% of all newly diagnosed mutant EGFR+ NSCLC cases.

[0657] Continuing with 15, we determined the unbound fraction in plasma and brain tissue (Table 2). 10 Both 15 and 1 showed similar plasma protein binding, with 1 (f u,脑 %=0.6), 15 showed excellent unbound rat brain tissue fraction (f u , 脑 Regarding potential toxicity, only moderate inhibition of the human ether-a-go-go related gene (hERG) potassium channel was observed with 15, IC 50 The maximum inhibition rate was 6.48 μM, and the maximum inhibition rate was about 60% at 10 μM (Table 2). We also tested the CYP inhibition of all major isomers (3A4, 1A2, 2C9, 2D6, 2C19), except for the moderate inhibition of CYP2D6 (IC 50 =1.1μM), no inhibitory effect was found (IC 5≥ 30M) (Table 2). Subsequent CYP2D6 time-dependent inhibition (TDI) IC in human liver microsomes and primary human hepatocytes 50 In excursion experiments, the activity of the latter was shown to be not time-dependent, eliminating concerns about potential drug-drug interactions (DDIs). 32 In the AMES fluctuation test with four Salmonella strains (TA98, TA100, TA1537, and TA1535) and in the in vitro micronucleus assay in Chinese hamster ovary (CHO-K1) cells with and without metabolic activation by rat liver S9 (+ / - S9), 15 was neither mutagenic nor genotoxic, contrary to the general belief that hydroxylamine itself is mutagenic and genotoxic (Table 2). 33-34，26，35 Together, these experiments demonstrate that 15 is a potent inhibitor of activation of cells carrying EGFR mutants and lacks the efflux propensity that characterizes currently approved targeted EGFR therapies.

[0658] Using KINOMEScan technology, we determined the selectivity of 15 against >400 human kinases at 1 μM concentration; an exquisite kinase selectivity of S(10) score of 0.015 was found (6 / 403 non-mutant kinases showed 10% activity at 1 μM ( Fig.10 ) 23 In addition to EGFR wt (0.30%), 15 showed high affinity for ERBB2 (HER2) (0.45%). Moderate affinity was also observed for ABL1 (7.7%), DRAK1 (8.3%), LYN (7.4%), and PIKFYVE (1.6%), but it was confirmed in subsequent binding experiments that considering the effective binding to EGFR (K d ≤0.2nM), which is of little significance ( Fig.11 As a result of the moderate affinity for HER2, this was observed in subsequent binding experiments (K d =21 nM), and considering the lack of effective treatment for metastatic HER2+ breast cancer 36 We further analyzed the effects of 15 on HER2 using additional biochemical and cellular assays. Fig.12 We found that 15 exhibited lower antiproliferative activity in HER2+ breast cancer cell lines compared with cells harboring EGFR+ mutations, with IC 50 Between 0.99 and 1.6 μM.

[0659] To identify 15 as a candidate for further investigation, we evaluated its pharmacokinetic properties in Sprague Dawley (SD) rats after administration ( Fig.13 ). Contrary to the common belief that hydroxylamine is inherently toxic, no adverse effects or signs of toxicity were observed for this compound at the doses tested. 33-34 In addition, SD rats showed high oral bioavailability (F = 64%), exposure (AUC inf =3907h·ng / mL) and an acceptable half-life (t 1 / 2 =2.37h), large distribution volume (V ss =6.67L / kg)( Fig.13 ). After a single oral dose of 20 mg / kg in SD rats, 15 showed a 0.33 Kp,uu (Partition coefficient of unbound brain to unbound plasma) (AUC inf ) and K of 0.95 p (brain-plasma partition coefficient) (AUC inf ), indicating excellent brain penetration ( Fig.14 ) 10，37 To confirm this effect, CD1 mice were given a single oral dose of 40 mg / kg and a K of 0.45 was obtained. p,uu (AUC inf ) and K of 0.77 p (AUC inf )( Fig.14 ).

[0660] Finally, to evaluate the feasibility of 15 as a potential treatment for BM in EGFR+ NSCLC, we used an intracranial patient-derived xenograft (PDX) model in which luciferase-labeled HCC827 cells were implanted into the brains of BALB / c nude mice ( Fig.15 At a dose of 10 mg / kg orally twice daily (bid), profound tumor regression was observed relative to the solvent control (1% methylcellulose) (P = 0.0059), confirming that 15 has intracranial antitumor activity ( Fig.16 ). In addition, within the 21-day treatment window, the average weight loss never exceeded 10%, and no adverse clinical reactions were observed at the study dose, thus demonstrating the potential of 15 as a drug candidate for CNS metastatic EGFR+ NNSCLC ( Fig.16 ).

[0661] in conclusion

[0662] We describe a highly selective, orally bioavailable, brain-penetrant EGFR inhibitor, 15, which contains a novel amine bioisostere, namely trisubstituted hydroxylamine. Contrary to common expectations for hydroxylamines in medicinal chemistry, 15 lacks mutagenicity or genotoxicity and exhibits good stability in vitro and in vivo. Intracranial PDX mouse models showed profound tumor regression after oral administration of 15, indicating that this novel compound is a potential lead for the treatment of local and CNS metastatic NSCLC driven by activating mutants of EGFR, as well as osimertinib-resistant EGFR+ NSCLC. In summary, our results suggest that trisubstituted hydroxylamine moieties are “structural alarms” in medicinal chemistry that can be incorporated into drug scaffolds to improve drug performance while maintaining potent bioactivity and avoiding molecular weight increase. These findings support the broader use of trisubstituted hydroxylamines as bioisosteres in drug development programs to optimize lead and patent lifecycle management.

[0663] It should be emphasized that the above-mentioned embodiments of the present disclosure are merely possible examples of embodiments proposed for a clear understanding of the principles of the present disclosure. Many changes and modifications may be made to the above-mentioned embodiments without substantially departing from the spirit and principles of the present disclosure. All such modifications and changes are intended to be included within the scope of the present disclosure and are protected by the following claims.

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Claims

1. A compound having structure I or a pharmaceutically acceptable salt thereof in R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; n is an integer from 1 to 5, wherein each R 2 are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, halide or alkoxy; m is an integer from 1 to 3, wherein each R 3 are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, halide or alkoxy; o is an integer from 1 to 10; X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino; R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and Each Z is independently hydrogen or deuterium.

2. The compound according to claim 1, wherein X is O.

3. The compound according to claim 1, wherein Y is O.

4. The compound according to claim 1, wherein Y is NR 5 , where R 5 It is a C1 to C5 alkyl group.

5. The compound according to claim 1, wherein Y is CR 6a R 6b , where R 6a is hydrogen, R 6b is a substituted or unsubstituted amino group.

6. The compound according to claim 1, wherein R 1 It's hydrogen.

7. The compound according to claim 1, wherein R 3 It is an alkoxy group.

8. The compound according to claim 1, wherein R 3 is an alkoxy group, and m is 1.

9. The compound according to claim 1, wherein o is an integer from 1 to 5.

10. The compound according to claim 1, wherein R 2 It is a halide, and n is 2.

11. The compound according to claim 1, wherein R 2 It is an ortho fluoride.

12. The compound of claim 1, wherein the compound has structure II or a pharmaceutically acceptable salt thereof in R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; R 2a and R 2b It is a halide; R 3 is an alkoxy group; o is an integer from 1 to 5; X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino; R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and Each Z is independently hydrogen or deuterium.

13. The compound according to claim 12, wherein X is O. The compound according to claim 12 , wherein Y is O.

15. The compound according to claim 12, wherein Y is NR 5 , where R 5 It is a C1 to C5 alkyl group.

16. The compound according to claim 12, wherein Y is CR 6a R 6b , where R 6a is hydrogen, and R 6b is a substituted or unsubstituted amino group.

17. The compound according to claim 12, wherein R 1 It's hydrogen.

18. The compound according to claim 12, wherein R 3 is a C1 to C10 substituted or unsubstituted straight or branched chain alkoxy group.

19. The compound according to claim 12, wherein R 3 It's a methoxy group.

20. The compound according to claim 12, wherein o is an integer from 1 to 5.

21. The compound according to claim 12, wherein R 2 It is a halide.

22. The compound according to claim 12, wherein R 2a is chloride, R 2b It's fluoride.

23. The compound according to claim 12, wherein R 2a is fluoride, R 2b It's chloride.

24. The compound according to claim 1, wherein the compound has structure III or a pharmaceutically acceptable salt thereof in R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; R 2a and R 2b It is a halide; R 3 is an alkoxy group; o is an integer from 1 to 5; X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and R 6a and R 6b are independently hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino; R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and Each Z is independently hydrogen or deuterium.

25. The compound according to claim 24, wherein X is O.

26. The compound of claim 24, wherein Y is O.

27. The compound according to claim 24, wherein Y is NR 5 , where R 5 It is a C1 to C5 alkyl group.

28. The compound according to claim 24, wherein Y is CR 6a R 6b , where R 6a is hydrogen, and R 6b is a substituted or unsubstituted amino group.

29. The compound according to claim 24, wherein R 1 It's hydrogen.

30. A compound according to claim 24, wherein R 3 is a C1 to C10 substituted or unsubstituted straight or branched chain alkoxy group.

31. The compound according to claim 24, wherein R 3 It's a methoxy group.

32. The compound of claim 24, wherein o is an integer from 1 to 5.

33. The compound according to claim 24, wherein R 2 It is a halide.

34. The compound according to claim 24, wherein R 2a is chloride, R 2b It's fluoride.

35. The compound according to claim 24, wherein R 2a is fluoride, R 2b It's chloride.

36. A compound according to claim 1, wherein the compound is 37. A pharmaceutical composition comprising a compound according to any one of claims 1 to 36 and a pharmaceutically acceptable carrier.

38. A method for treating a subject having non-small cell lung cancer, neuroblastoma, glioblastoma multiforme, metastatic brain cancer, brain cancer, prostate cancer or breast cancer, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 36.

39. A method of treating a subject having osimertinib-resistant cancer, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 36.

40. A method of treating a subject having brain metastatic cancer, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 36.

41. A method of inhibiting epidermal growth factor receptor (EGFR) in a subject, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 36.

42. The method of claim 38, wherein the compound is administered orally to the subject.

43. The method of claim 39, wherein the compound is administered orally to the subject.

44. The method of claim 40, wherein the compound is administered orally to the subject.

45. The method of claim 38, wherein the compound is administered at a dosage of about 50 mg per day to about 1000 mg per day.

46. ​​The method of claim 39, wherein the compound is administered at a dosage of about 50 mg per day to about 1000 mg per day.

47. The method of claim 40, wherein the compound is administered at a dosage of about 50 mg per day to about 1000 mg per day.

48. A method for preparing a compound as claimed in any one of claims 1 to 16, the method comprising reacting a compound having structure IV with a compound having structure V in the presence of a base in R 1 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; R 2a and R 2b It is a halide; R 3 is an alkoxy group; o is an integer from 1 to 5; X is O or NR 4 , where R 4 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl; Y is O, NR 5 or CR 6a R 6b , where R 5 is hydrogen, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, or substituted or unsubstituted aryl, and R 6a and R 6b are independently hydrogen, deuterium, substituted or unsubstituted straight or branched chain alkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino; R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; each Z is independently hydrogen or deuterium; and LG is a leaving group.

49. The method of claim 48, wherein LG is a halogen or a sulfonic acid group.

50. The method of claim 48, wherein the base comprises a hydride, an alkoxide, a Grignard reagent, or an alkyl lithium compound.

51. A method for preparing a compound having structure X in R 5 is a substituted or unsubstituted straight chain or branched chain alkyl group, and R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and each Z is independently hydrogen or deuterium, The method comprises (a) reacting a compound having structure XI with a compound having structure XII in the presence of a base to produce a compound having structure XIII Where R 10 is a substituted or unsubstituted straight chain or branched chain alkyl group, or a substituted or unsubstituted straight chain or branched chain alkoxy group, and LG is the leaving group; (b) reacting a compound having structure XIII with a first oxidant under heating to produce a compound having structure XIV (c) reacting a compound having structure XIV with (i) a second oxidizing agent and then with (ii) a first reducing agent to produce a first intermediate; and (d) reacting the first intermediate with a second reducing agent to produce a compound having structure X.

52. The method of claim 51, wherein R 10 It is a C1 to C5 straight or branched chain alkyl group.

53. The method of claim 51, wherein R 10 It is a C1 to C5 straight chain or branched chain alkoxy group.

54. The method of claim 51, wherein R 10 It is a C1 to C5 straight or branched chain alkoxy group substituted with an aryl group.

55. The method of claim 51, wherein R 10 It is benzyloxy.

56. The method of claim 51, wherein LG is a halide, sulfonate, carbonate or phosphate.

57. The process of claim 51, wherein the base in step (a) comprises a carbonate, a hydroxide, a phosphate, a hydride, a dialkylamide or hexamethyldisilazane.

58. The method of claim 51, wherein step (a) is performed in an aprotic organic solvent.

59. The method of claim 51, wherein step (a) is performed at a temperature of about 25°C to about 100°C.

60. The method of claim 51, wherein step (b) comprises the steps of: (i) reacting a compound having structure XIII with a first oxidizing agent in a first organic solvent to prepare a first composition; (ii) adding an aqueous base to the first composition to prepare a second composition comprising an organic layer and an aqueous layer; (iii) separating the organic layer from the aqueous layer; (iv) removing the first organic solvent from the organic layer to produce a residue; (v) dissolving the residue in a second organic solvent to prepare a second composition; as well as (vi) heating the second composition from about 50°C to about 100°C to produce a compound having structure XIV.

61. The method of claim 60, wherein the first organic solvent is dichloromethane and the second organic solvent is toluene.

62. The method of claim 51, wherein the first oxidizing agent in step (b) comprises a peroxyacid, ozone, or hydrogen peroxide / acetic acid.

63. The method of claim 51, wherein the first oxidizing agent in step (b) comprises meta-chloroperbenzoic acid.

64. The method of claim 51, wherein the molar ratio of the first oxidizing agent to the compound of structure XIII is from 0.95:1 to 1:1.

05.

65. The method of claim 51, wherein step (c) comprises the steps of: (i) reacting a compound having structure XIV with a second oxidizing agent in a third organic solvent to produce a third composition; and (ii) mixing a first reducing agent with a third composition to prepare a first intermediate.

66. The method of claim 65, wherein the second oxidant comprises ozone or osmium tetroxide and sodium periodate.

67. The method of claim 65, wherein the third organic solvent comprises an alcohol and an aprotic solvent.

68. The method of claim 65, wherein the compound of structure XIV is reacted with the second oxidant at a temperature of about -50°C to about -100°C.

69. The method of claim 51, wherein the first reducing agent comprises a hydride.

70. The method of claim 51, wherein the first reducing agent comprises a borohydride.

71. The method of claim 51, wherein the molar ratio of the first reducing agent to the compound of structure XIV is from 1.5:1 to 2.5:

1.

72. The method of claim 51, wherein the first intermediate is isolated prior to step (d).

73. The method of claim 51, wherein step (d) comprises the steps of: (i) dissolving the first intermediate in an aprotic solvent to prepare a fourth composition; (ii) mixing the second reducing agent with the fourth composition to prepare a compound having structure X; and (iii) isolating and purifying the compound having structure X.

74. The method of claim 73, wherein the second reducing agent is mixed with the fourth composition at a temperature of about 10°C to about -50°C.

75. The method of claim 51, wherein the second reducing agent comprises a hydride.

76. The method of claim 51, wherein the second reducing agent comprises aluminum hydride.

77. The method of claim 51, wherein the molar ratio of the second reducing agent to the first intermediate is 4:1 to 2:

1.

78. The method of claim 51, wherein R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are hydrogen respectively.

79. The method of claim 51, wherein R 5 is methyl, R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are hydrogen respectively.

80. A method for preparing a compound having structure XX in R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and each Z is independently hydrogen or deuterium, The method comprises (d) reacting a compound having structure XXI with a compound having structure XXII in the presence of a base to prepare a compound having structure XXIII Where LG is the leaving group; (a) reacting a compound having structure XXIII with a first oxidant to produce a compound having structure XXIV as well as (b) reacting a compound having structure XXIV with (i) a second oxidizing agent and then with (ii) a first reducing agent to produce a compound having structure XX.

81. The method of claim 80, wherein LG is a halide, sulfonate, carbonate or phosphate.

82. The process of claim 80, wherein the base in step (a) comprises a carbonate, a hydroxide, a phosphate, a hydride, a dialkylamide, or hexamethyldisilazane.

83. The method of claim 80, wherein step (a) is performed in an aprotic organic solvent.

84. The method of claim 80, wherein step (a) is performed at a temperature of about 25°C to about 100°C.

85. The method of claim 80, wherein step (b) comprises the steps of: (i) reacting a compound having structure XXIII with a first oxidizing agent in a first organic solvent to prepare a first composition; (ii) adding an aqueous base to the first composition to prepare a second composition comprising an organic layer and an aqueous layer; (iii) separating the organic layer from the aqueous layer; (iv) removing the first organic solvent from the organic layer to produce a residue; (v) dissolving the residue in a second organic solvent to prepare a second composition; and (vi) heating the second composition from about 50°C to about 100°C to produce a compound having structure XXIV.

86. The method of claim 85, wherein the first organic solvent is dichloromethane and the second organic solvent is toluene.

87. The method of claim 80, wherein the first oxidizing agent in step (b) comprises a peroxyacid, ozone, or hydrogen peroxide / acetic acid.

88. The method of claim 80, wherein the first oxidizing agent in step (b) comprises meta-chloroperbenzoic acid.

89. The method of claim 80, wherein the molar ratio of the first oxidizing agent to the compound of structure XIII is from 0.95:1 to 1:1.

05.

90. The method of claim 80, wherein step (c) comprises the steps of: (i) reacting a compound having structure XXIV with a second oxidizing agent in a third organic solvent to produce a third composition; and (ii) mixing a first reducing agent with the third composition to prepare a compound having structure XX; and (iii) isolating and purifying the compound having structure XX.

91. The method of claim 90, wherein the second oxidant comprises ozone or osmium tetroxide and sodium periodate.

92. The method of claim 90, wherein the third organic solvent comprises an alcohol and an aprotic solvent.

93. The method of claim 90, wherein the compound of structure XXIV is reacted with the second oxidant at a temperature of about -50°C to about -100°C.

94. The method of claim 80, wherein the first reducing agent comprises a hydride.

95. The method of claim 80, wherein the first reducing agent comprises a borohydride.

96. The method of claim 80, wherein the molar ratio of the first reducing agent to the compound of structure XXIV is from 1.5:1 to 2.5:

1.

97. The method of claim 80, wherein R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are hydrogen respectively.

98. The method of claim 51, wherein R 5 is methyl, R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are hydrogen respectively.

99. The method of claim 80, wherein R 5 is methyl, R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are hydrogen respectively.

100. A compound having structure XXX in Y is O or NR 5 , R 5 is a substituted or unsubstituted straight chain or branched chain alkyl group, and R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently hydrogen, deuterium, or substituted or unsubstituted alkyl; and Each Z is independently hydrogen or deuterium.

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