Therapeutic conjugate

Abstract

We provide therapeutic conjugates that can covalently bind to biological targets. [Solution] A compound having the formula: (FCB)a-(L)b-(CLM)c or a pharmaceutically acceptable salt thereof, wherein CLM is, for example JPEG2026090283000084.jpg20170 And FCB, for example, JPEG2026090283000085.jpg48170 A compound in which L is, for example, an unsubstituted C5-C6 heterocycloalkyl group.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 902,554, titled THERAPEUTIC CONJUGATES, filed on 19 September 2019, and U.S. Provisional Patent Application No. 63 / 078,055, also titled THERAPEUTIC CONJUGATES, filed on 14 September 2020, both of which are incorporated herein by reference.

[0002] This disclosure generally relates to therapeutic conjugates that covalently bind to biological targets. [Background technology]

[0003] Covalent inhibitors bind to receptors in the same way as classical inhibitors, but instead of dissociating, they form a permanent covalent chemical bond to the receptor. Some examples of covalent inhibitors include penicillin, aspirin, clopidogrel, the EGFR kinase inhibitor afatinib used to treat lung cancer, and the Bruton's tyrosine kinase inhibitor ibrutinib used to treat B-cell malignancies. Furthermore, in the field of oncology, covalent inhibitors have been shown to be effective against drug-resistant tumors and generally exhibit a stronger ability to inhibit tumor growth.

[0004] In recent years, covalent inhibitors have attracted the attention of major pharmaceutical companies because they offer higher potential and longer duration of action compared to classical reversible inhibitors. The extended duration of action means that lower dosing frequency is required; that is, patients need to take fewer tablets less frequently.

[0005] There is a need to design therapeutic conjugates that can covalently bind to biological targets and to develop high-throughput screening methods for these therapeutic conjugates. [Overview of the project]

[0006] In some embodiments, the disclosure provides therapeutic conjugates that can form a covalent bond with a kinase or pseudokinase. The kinase may be PI3 kinase (PI3K). The therapeutic conjugate may have the structure (FCB)a-(L)b-(CLM)c, where a and c are independently integers from 1 to 5, and b is an integer from 0 to 5, and the FCB portion comprises a PI3K inhibitor or a fragment, analog, or derivative thereof. In some embodiments, the FCB is [ka] The therapeutic conjugate may contain a structure selected from the group consisting of compounds 1-101 to 1-172.

[0007] In some embodiments, the therapeutic conjugate is [ka] It may have the structure or a pharmaceutically acceptable salt thereof, and L is [ka] Selected from the group consisting of, either end can be connected to the CLM, and R1 is [ka] Selected from the group consisting of, CLM is [ka] The therapeutic conjugate is selected from the group consisting of compounds 1-101, 1-102, 1-103, 1-104, 1-105, 1-106, 1-107, 1-108, 1-109, 1-110, 1-111, 1-112, 1-113, 1-114, 1-115, 1-116, 1-117, 1-118, 1-119, 1-120, 1-121, 1-122, You may select from the group consisting of 1-123, 1-124, 1-125, 1-126, 1-127, 1-128, 1-129, 1-130, 1-137, 1-138, 1-145, 1-146, 1-153, 1-154, 1-161, 1-162, 1-169, 1-170, 1-171, and 1-172.

[0008] In some embodiments, the therapeutic conjugate is [ka] It may have the structure or a pharmaceutically acceptable salt thereof, and L is [ka] And R2 is [ka] Selected from the group consisting of, CLM is [ka] The therapeutic conjugate may be selected from the group consisting of compounds 1-131, 1-132, 1-133, 1-134, 1-135, 1-136, 1-139, 1-140, 1-141, 1-142, 1-143, 1-144, 1-147, 1-148, 1-149, 1-150, 1-151, 1-152, 1-155, 1-156, 1-157, 1-158, 1-159, 1-160, 1-163, 1-164, 1-165, 1-166, 1-167, and 1-168.

[0009] In some embodiments, the therapeutic conjugate may contain a structure selected from compounds 1-101 to 1-172 or a pharmaceutically acceptable salt thereof. In some embodiments, the therapeutic conjugate may contain a structure selected from compounds 1-1 to 1-11 or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the Disclosure provides a pharmaceutical composition which may comprise a therapeutic conjugate disclosed herein and at least one pharmaceutically acceptable excipient.

[0011] In some embodiments, the Disclosure provides methods for modulating the activity of a kinase or pseudokinase, comprising administering a therapeutic conjugate disclosed herein. In some embodiments, the activity of a kinase or pseudokinase can be inhibited. In some embodiments, the kinase may be PI3-K.

[0012] In some embodiments, the Disclosure provides a method for treating a subject in need of treatment, the method comprising administering a therapeutically effective amount of a pharmaceutical composition described herein. The subject may have a treatment condition selected from the group consisting of cancer, neurodegenerative diseases, autoimmune disorders, and aging. In some embodiments, the subject may have cancer. In some embodiments, the subject may have cancer with a mutation in the PIK3CA gene. [Modes for carrying out the invention]

[0013] I. Composition The inventors, in particular, discovered the Anchored Relational Covalent System, hereafter referred to as ARCS, which includes a Functionally Competent Binder, hereafter referred to as FCB, and a Covalent Linking Modality, hereafter referred to as CLM, where CLM is directly or indirectly coupled to a therapeutic modality, is optional, and is positioned between FCB and CLM. Includes a linker. In some embodiments, the CLM is directly and covalently bonded to the FCB by a linker. In some embodiments, the CLM is indirectly and covalently bonded to the FCB by a linker.

[0014] As used herein, the term "ARCS" refers to any therapeutic conjugate formed by linking FCB and CLM by a linker or by binding. In some embodiments, ARCS can form covalent bonds with one or more targets, such as nucleotides, oligonucleotides, peptides, or proteins. In some embodiments, ARCS can form covalent bonds with biological targets. Covalent bonds can be detected using any known method in the art. As an unrestricted example, the covalent bond of an azide small molecule to a protein can be detected by using click chemistry to bind a heavy PEG-containing alkyne to the small molecule. Covalently labeled proteins are PEG-labeled and have a higher molecular weight, and are therefore detected by gel shift (Biochemistry 2018, 57:5769-5774). In another unrestricted example, covalently labeled purified proteins can be detected using mass spectrometry (Nature Chemical Biology 2007, 3:229-238). In yet another non-limiting example, covalent bonding can be analyzed using proteomics methods based on quantitative mass spectrometry of cells (Cell Chemical Biology 2017, 24:1388-1400.e7). In yet another non-limiting example, covalent bond formation can be confirmed using X-ray crystallography (Nature Chemical Biology 2007:3:229-238; J.Med.Chem.2020, 63:52-65). In yet another non-limiting example, the site of covalent modification can be revealed using mass spectrometry of intracellular, covalently labeled, and affinity-rich samples (Nat.Chem.Biol.2016, 12:876-884).

[0015] In some embodiments, ARCS can form covalent bonds with biological targets in proportions ranging from approximately 5% to 100% of the biological target. In some embodiments, ARCS can form covalent bonds with biological targets in proportions ranging from approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the covalent bonds are formed in an aqueous solution at a therapeutic dose of 10 mM within 48 hours at a temperature of 0–50°C.

[0016] While we do not wish to be bound by any theory, ARCS can first form non-covalent bonds with biological targets (such as target proteins) via FCBs, and then covalent bonds with biological targets via CLMs. In some embodiments, the efficacy of ARCS is superior to that of FCBs alone. In some embodiments, CLMs do not substantially interfere with the efficacy of FCBs. In some embodiments, FCBs do not substantially interfere with the covalent bonding of CLMs. In some embodiments, the toxicity of ARCS is less than that of FCBs alone.

[0017] As used herein, the term “toxicity” refers to the ability of a substance or composition to be harmful or toxic to cells, living tissues, or the cellular environment. “Low toxicity” refers to the attenuated ability of a substance or composition to be harmful or toxic to cells, living tissues, or the cellular environment. Such attenuation or low toxicity may be relative to a standard measure, to treatment, or to the absence of treatment.

[0018] As used herein, the term "FCB" refers to a therapeutic modality which may be a known drug, diagnostic compound, drug candidate and functional fragment and / or any combination thereof. FCBs are free acid and free base forms, optical isomers and tautomers, and drugs The FCB comprises isotopes including radioisotopes and pharmaceutically acceptable salts, their prodrugs or fragments. The FCB may be a small molecule, protein, peptide, lipid, carbohydrate, sugar, nucleic acid, or a combination thereof. In some embodiments, the FCB is a nucleic acid including, but not limited to, DNA or RNA. The FCB may be a therapeutic agent, such as an anticancer agent, an antineurodegenerative agent, an autoimmune agent, or an anti-aging agent, but is not limited. The FCB may be bound to a biological target non-covalently. In some embodiments, the FCB may be a functional fragment of a drug. As used herein, the term “functional fragment” refers to a part of a drug or a derivative or analogue that can induce the desired effect of the drug. In some embodiments, the FCB may contain an alkyne functional group. In some embodiments, the FCB may not contain an alkyne functional group.

[0019] As used herein, “peptide,” “polypeptide,” and “protein” refer to polymers composed of amino acid monomers linked by amide bonds. The amino acids may be D or L optical isomers. Peptides may be formed by condensation or coupling reactions between the α-carbon carboxyl group of one amino acid and the amino group of another. Peptides may be nonlinear branched peptides or cyclic peptides. Furthermore, peptides may be optionally modified or protected with a wide range of functional or protecting groups, including amino and / or carboxyl termini.

[0020] The amino acid residues of peptides are abbreviated as follows: Phenylalanine is Phe or F, leucine is Leu or L, isoleucine is Ile or I, methionine is Met or M, valine is Val or V, serine is Ser or S, proline is Pro or P, threonine is Thr or T, alanine is Ala or a, tyrosine is Tyr or Y, histidine is His or H, glutamine is Gln or Q, asparagine is Asn or N, lysine is Lys or K, aspartic acid is Asp or D, glutamic acid is Glu or E, cysteine ​​is Cys or C, tryptophan is Trp or W, arginine is Arg or R, and glycine is Gly or G.

[0021] As used herein, the term "CLM" refers to any covalent modality capable of forming a covalent bond with a biological target. The CLM may be linked to the FCB by a bond or a linker. The CLM may contain one or more chemical moieties capable of forming a covalent bond with a biological target. The chemical moieties may be electrophilic or nucleophilic groups.

[0022] CLM may be small molecules having molecular weights of less than approximately 1,000 Da, less than approximately 900 Da, less than approximately 800 Da, less than approximately 700 Da, less than approximately 600 Da, or less than approximately 500 Da. In some cases, CLM may have molecular weights of approximately 5 Da to approximately 1,000 Da, approximately 10 Da to approximately 900 Da, approximately 20 Da to approximately 700 Da in some embodiments, approximately 20 Da to approximately 500 Da in some embodiments, approximately 50 Da to approximately 400 Da, approximately 100 Da to approximately 300 Da in some embodiments, and approximately 150 Da to approximately 300 Da in some embodiments. The molecular weight of CLM can be calculated by multiplying the sum of the atomic weights of each atom in the formula of CLM by the number of each atom. It can also be measured by mass spectrometry, NMR, chromatography, light scattering, viscosity, and / or any other method known in the art. The unit of molecular weight may be g / mol, dalton (Da), or atomic mass unit (amu), and it is known that 1 g / mol = 1 Da = 1 amu.

[0023] As used herein, the term “biological target” refers to any target to which FCBs bind non-covalently to produce a therapeutic effect. CLMs bind covalently to biological targets. It is bound to a target. In some embodiments, the biological target is a protein. Non-limiting examples of biological targets include kinases such as phosphoinositide 3-kinase (PI3K) and pseudokinase, but are not limited to the following.

[0024] In some embodiments, ARCS can form covalent bonds with PI3 kinase. In some embodiments, ARCS can form covalent bonds with PI3 kinase from about 5% to 100% of PI3 kinase. In some embodiments, ARCS can form covalent bonds with PI3 kinase from about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of PI3 kinase.

[0025] ARCS comprises at least one FCB optionally coupled to at least one CLM by a linker. In some embodiments, ARCS may be a therapeutic conjugate between a single FCB and a single CLM, for example having the structure XLY, where X is a CLM, L is an optional linker, and Y is an FCB. In some embodiments, ARCS may be a therapeutic conjugate between a single therapeutic modality and a single covalent modality. In some embodiments, X is a covalent modality, L is an optional linker, and Y is a therapeutic modality.

[0026] In some embodiments, the ARCS includes one or more FCBs, one or more linkers, one or more CLMs, or any combination thereof. The ARCS may have any number of FCBs, linkers, and CLMs. The ARCS may also include, but are not limited to, XLYLX, (XLY) n ,YLXLY,X-(LY) n (XL) n -Y, (X) n -LY or XL-(Y) n ARCS can have the following structure, where L is an optional linker, Y is an FCB, and n is an integer between 2 and 100, 2 and 50, 2 and 20, for example, between 2 and 5. Each occurrence of X, L, and Y may be the same or different, for example, ARCS may include one or more types of FCBs, one or more types of linkers, and / or one or more types of CLMs.

[0027] In some embodiments, the ARCS may contain one or more types of CLMs bonded to a single FCB. For example, the ARCS may contain a single FCB having multiple CLMs, each bonded via the same or different linkers. The ARCS may have the structure XLYLX, where each X is a CLM, which may be the same or different, each L is a linker, which may be the same or different, and Y is an FCB.

[0028] In some embodiments, the ARCS may contain one or more CLMs attached to a single FCB. For example, the ARCS may include one CLM having a plurality of FCBs attached thereto via the same or different linkers. The ARCS may have a structure Y-L-X-L-Y, where X is a CLM, each L is a linker that may be the same or different, and Y is an FCB that may be the same or different.

[0029] In some embodiments, the ARCS is a therapeutic conjugate, and the therapeutic conjugate a. includes a therapeutic modality, which is selected from the group consisting of one or more of known drugs, diagnostic compounds, drug candidates, and functional fragments and / or any combination of the foregoing, b. includes a covalent bonding modality, which includes one or more chemical moieties, one or more of which can form a covalent bond with a biological target, and this covalent bonding mo dality is directly or indirectly attached to this therapeutic modality, and c. optionally includes a linker positioned between the therapeutic modality and the covalent bonding modality.

[0030] In some embodiments, the therapeutic conjugate includes a formula selected from the group consisting of a) X-L-Y, b) X-L-Y-L-X, c) (X-L-Y) n , d) Y-L-X-L-Y, e) X-(L-Y) n , f) (X-L) n -Y, g) (X) n -L-Y and h) X-L-(Y) n ; X is a covalent bonding modality, L is an optional linker, Y is a therapeutic modality, and n is an integer between 2 and 100.

[0031] The purpose of this disclosure is to design methods for synthesizing ARCS and its compositions, as well as ARCS and libraries of ARCS.

[0032] Furthermore, an objective of this disclosure is to provide a method for screening the ARCS library to identify candidate covalent bindings to biological targets.

[0033] A further object of this disclosure is to provide a method for administering ARCS and its compositions to subjects requiring them.

[0034] A.FCB The ARCS of this disclosure contains at least one FCB. The ARCS of this disclosure may contain one or more FCBs, which may be the same or different. An FCB may be a therapeutic modality that affects any biological process and is used to prevent, diagnose, alleviate, treat or cure a disease state. An FCB may be a therapeutic agent, prophylactic agent, diagnostic agent or nutritional agent. The efficacy of an FCB or ARCS refers to the efficacy of the FCB or ARCS for its intended purpose, i.e., the ability of a given FCB or ARCS to produce a desired pharmacological effect. As used herein, the term “pharmacological activity” means activity that modulates or modulates a biological process to result in a phenotypic change, such as cell death or reduced cell proliferation.

[0035] In some embodiments, FCB is a PI3 kinase inhibitor. In some embodiments, FCB is a pyrrolo[2,1-F[1,2,4]triazine compound. In some embodiments, FCB is a PI3 kinase inhibitor having one of the formulas described in U.S. Patent No. 9,724,352B2, the entire contents of which are incorporated herein by reference. In some embodiments, FCB is one of the compounds shown in Table 1 of U.S. Patent No. 9,724,352B2. In some embodiments, FCB is [ka] It includes the structure.

[0036] In some embodiments, the FCB is structure [ka] It is a compound having, and R1 is [ka] It is selected from the group consisting of the following.

[0037] In some embodiments, the FCB is structure [ka] It is a compound having, and R2 is, [ka] It is selected from the group consisting of the following.

[0038] Generally, the efficacy of FCBs is achieved by non-covalent binding to biological targets. Non-covalent binding is achieved by a certain degree of specificity and / or affinity to the target. While both specificity and affinity are generally desirable, in certain cases higher specificity can compensate for lower affinity, and higher affinity for lower specificity. The requirements for affinity and specificity vary depending on various factors, but are not limited, including the absolute concentration of the target, the relative concentration of the target (e.g., in cancer vs. normal cells), potential and toxicity, the route of administration, and / or diffusion or transport to target cells. At the molecular or cellular level, the effects of FCBs (ARCS or alone) may include, but are not limited, enhancement or inhibition of target activity, labeling of the target, and / or alteration of target cells (e.g., cell death).

[0039] In some embodiments, FCB may be a small molecule, protein, peptide, lipid, carbohydrate, sugar, nucleic acid, or a combination thereof. In some embodiments, FCB may be a therapeutic agent, but is not limited to, an anticancer agent, an antineurodegenerative agent, an autoimmune agent, and an anti-aging agent. Various therapeutic agents are well known in the art and may be used in the compositions described herein.

[0040] In some embodiments, FCB is a small molecule. In some embodiments, FCB may be a protein, peptide, or nucleic acid. In some embodiments, FCB may be a lipid. In some embodiments, FCB may be a carbohydrate or sugar. In some embodiments, FCB has an alkyne group. In some embodiments, FCB may not have an alkyne functional group.

[0041] In some embodiments, the FCB may be a functional fragment of the drug. As used herein, the terms “functional fragment” or “drug core” refer to a portion of the drug or a derivative or analogue of the drug that can induce the desired effect of the drug.

[0042] In some embodiments, the FCB may be bound to a biological target non-covalently. In some embodiments, the FCB is IC with a thickness of <1000 μm, 900 μm, 800 μm, 700 μm, 600 μm, or 500 μm. 50 It may also be bound to a biological target.

[0043] In some embodiments, FCB is an anticancer agent. In some embodiments, FCB is an antineurodegenerative agent. In some embodiments, FCB is an autoimmune agent. In some embodiments, FCB is an anti-aging agent.

[0044] In certain embodiments, the FCB of ARCS is the molar weight portion of the components of ARCS. The ARCS contains a predetermined molar weight percentage of approximately 1% to 10%, or approximately 10% to 20%, or approximately 20% to 30%, or approximately 30% to 40%, or approximately 40% to 50%, or approximately 50% to 60%, or approximately 60% to 70%, or approximately 70% to 80%, or approximately 80% to 90%, or approximately 90% to 99%, such that the sum of the ions equals 100%. The amount of FCB in ARCS may be expressed as a percentage of CLM. For example, this teaching provides ratios of FCB to CLM of approximately 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

[0045] B.CLM The ARCS of this disclosure comprises one or more CLMs. A CLM can be any covalent modality capable of forming a covalent bond with a biological target. A CLM may comprise one or more chemical moieties, one or more chemical moieties capable of forming a covalent bond with a biological target. In certain embodiments, a CLM may comprise an internal linker or spacer. The internal linker or spacer may be a combination of two parts of the CLM, or can be bonded to the CLM.

[0046] In some embodiments, CLM is a small molecule. In some embodiments, CLM has a molecular weight of less than about 1000 daltons (e.g., less than about 900, 800, 750, 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, 100, etc.).

[0047] In certain embodiments, the CLM of ARCS contains a predetermined molar weight percentage of about 1% to about 10%, or about 10% to about 20%, or about 20% to about 30%, or about 30% to about 40%, or about 40% to about 50%, or about 50% to about 60%, or about 60% to about 70%, or about 70% to about 80%, or about 80% to about 90%, or about 90% to about 99%, such that the sum of the molar weight percentages of the components of ARCS equals 100%. The amount of CLM of ARCS may be expressed as a percentage of the FCB. For example, this instruction provides FCB ratios to CLM of approximately 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

[0048] In some embodiments, CLM comprises at least one substituted or unsubstituted alkyne. In some embodiments, CLM comprises at least one substituted or unsubstituted acrylamide. In some embodiments, CLM comprises at least one substituted or unsubstituted vinylsulfonamide. In some embodiments, CLM comprises at least one substituted or unsubstituted vinylsulfone. In some embodiments, CLM comprises at least one substituted or unsubstituted fumaamide. In some embodiments, CLM comprises at least one substituted or unsubstituted acrylic acid. In some embodiments, CLM comprises at least one substituted or unsubstituted isothiocyanate. In some embodiments, CLM comprises at least one substituted or unsubstituted sulfonyl fluoride. In some embodiments, CLM comprises at least one substituted or unsubstituted fluorosulfate. In some embodiments, CLM comprises at least one substituted or unsubstituted formylphenylboronic acid. In some embodiments, CLM comprises at least one substituted or unsubstituted boronic acid. In some embodiments, CLM comprises at least one activated ester. In some embodiments, CLM comprises at least one substituted or unsubstituted thioester. In some embodiments, CLM comprises at least one sulfonyl group. In some embodiments, CLM comprises at least one nitro group. In some embodiments, CLM comprises at least one substituted or unsubstituted epoxide. In some embodiments, CLM comprises at least one substituted or unsubstituted formylphenylboronic acid. In some embodiments, CLM comprises at least It contains one substituted or unsubstituted aryl halide. In some embodiments, the CLM contains at least one substituted or unsubstituted aldehyde. In some embodiments, the CLM contains at least one substituted or unsubstituted triazine. In some embodiments, the CLM contains at least one substituted or unsubstituted cyanoacrylamide. In some embodiments, the CLM contains at least one substituted or unsubstituted chloroacetamide.

[0049] Examples of CLMs, though not limited to these, include the following: [ka] [ka] [ka] [ka] [ka] A, B, C, and D appear independently in each instance as H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, any substituent for A, B, C, and D can be independently halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, or optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups.

[0050] A1, A2, A3, A4, A5, and A6 appear independently as H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, A 1、 A2, A 3、 A 4、 The optional substituents of A5 and A6 are, independently, halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on a cycloalkyl group are one to two substituents, independently of halogen, OH, NH2, or CH. 3、 The group is selected from CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3 or their fragments, derivatives, or analogs.

[0051] In some embodiments, CLM [ka] It is selected from the group consisting of the following.

[0052] C. Linker The ARCS of this disclosure includes one or more optional linkers connecting the FCB and CLM. The linker, L, can be coupled anywhere on the FCB and CLM, as long as the effectiveness of the FCB and the coupling of the CLM are not significantly affected. In some embodiments, the CLM includes an optional internal linker.

[0053] In some embodiments, the linker (including the internal linker of the CLM) is a small molecule. In some embodiments, the linker (including the internal linker of the CLM) is selected, but is not limited to, substituted and unsubstituted C1-C. 30 Alkyl, substituted, and unsubstituted C2-C 30 Alkenyl, substituted and unsubstituted C2-C 30 Alkynyl, substituted and unsubstituted C3-C 30 Cycloalkyl, substituted and unsubstituted C1-C 30 Heterocycloalkyl, substituted and unsubstituted C3-C 30 Cycloalkenyls, substituted and unsubstituted C1-C 30 This includes heterocycloalkenyls, substituted and unsubstituted aryls, and substituted and unsubstituted heteroaryls.

[0054] In some embodiments, the linker (including the internal linker of the CLM) is C1-C 10 Linear alkyl, C1-C 10 Linear O-alkyl, C1-C 10 Linear substituted alkyl, C1-C 10 Linear substituted O-alkyl, C4-C 13 Branched-chain alkyl, C 4- C 13 Branched chain O-alkyl, C2-C 12 Linear alkenyl, C2-C 12 Linear O-alkenyl, C 3- C 12 Linear substituted alkenyl, C3-C 12Linear substituted O-alkenyls, polyethylene glycol, polylactic acid, polyglycolic acid, poly(lactide-co-glycolide), polycarboractone, polycyanoacrylate, ketone, aryl, heterocyclic, succinate ester, amino acid, aromatic group, ether, crown ether, urea, thiourea, amide, purine, pyrimidine, bipyridine, indole derivatives acting as crosslinkers, chelating agents, aldehydes, ketones, bisamines, bisalcohols, heterocyclic structures, aziline, disulfide, thioether, hydrazone, and combinations thereof. For example, the linker may be a C3 linear alkyl or ketone. The alkyl chain of the linker may have one or more components. The linker may be substituted with a substitution group or heteroatom. In some embodiments, the linker contains one or more atoms or groups selected from -O-, -C(=O)-, -NR, -OC(=O)-NR-, -S-, and -SS-. The linker may be selected from dicarboxylate derivatives of succinic acid, glutamic acid, or diglycolic acid.

[0055] In some embodiments, the alkyl chain of the linker may be optionally interrupted by one or more atoms or groups selected from -O-, -C(=O)-, -NR, -OC(=O)-NR-, -S-, and -SS-. The linker may be selected from dicarboxylate derivatives of succinic acid, glutamic acid, or diglycolic acid.

[0056] In some embodiments, the linker may be non-cleavable. In some embodiments, the linker may be cleavable. In some embodiments, the linker may be cleaved by an enzyme.

[0057] As an unrestricted example of a linker, [ka] These include, [ka] D1, D2, D3, D4, D5, and D6 are independently selected in each occurrence from the group consisting of N, C, O, or S, and D 1-6 If it is N, the corresponding position is trivalent, D 1-6 If it is O or S, the corresponding position is divalent, B1, B2, B3, B 4、 B5 and B6 are either absent in each occurrence or independently H, halogen, CF3, -OH, -CH3, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1~3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH 3) OCO(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 The optional substituents for B1, B2, B3, and B4 are selected from cycloalkyl groups, optionally substituted 5-10 membered heterocycles, optionally substituted aryl groups, and optionally substituted 5-10 membered heteroaryl groups, and the optional substituents for B1, B2, B3, and B4 are independently halogens, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1~3 Alkyl and optionally substituted C 3-6 One to three substituents selected from the group consisting of cycloalkyl groups, C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogens. -N, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH and -S-CH 3、or one or two substituents selected from the group consisting of any fragment or analogue thereof.

[0058] In some embodiments, the linker is [ka] A selection is made from the group consisting of, and either end can be connected to the CLM. In some embodiments, [ka] The linker selected from the group consisting of the following is: [ka] It is connected to a CLM selected from the group consisting of the following.

[0059] D.ARCS The ARCS of this disclosure represent a class of drugs that offer many advantages compared to reversible inhibitors, including increased potential and extended duration of action. This disclosure provides therapeutic conjugates that form a covalent bond with a kinase or pseudokinase. In some embodiments, the kinase is PI3 kinase (PI3K). The therapeutic conjugate is It may also have the structure (FCB)a-(L)b-(CLM)c. a and c are independent integers between 1 and 5. b is an integer between 0 and 5. The FCB portion contains a PI3K inhibitor or a fragment, analog, or derivative thereof.

[0060] The FCB, L (linker), and CLM parts are explained in the section above. In one non-exclusive example, the FCB is... [ka] Includes.

[0061] In some embodiments, the FCB is structure [ka] It is a compound having, and R1 is [ka] It is selected from the group consisting of the following.

[0062] In some embodiments, the FCB is structure [ka] It is a compound having, and R2 is, [ka] Selected from the group consisting of: [ka] A selection from the group consisting of, and either end can be connected to the CLM. In some embodiments, the CLM is [ka] It is selected from the group consisting of the following.

[0063] In some embodiments, ARCS is a broad general structural compound 1-1 to compound 1-5. [ka] or selected from the group consisting of pharmaceutically acceptable salts thereof, where R1 is independently in each occurrence. [ka] Selected from the group consisting of, R1 can bind to a functional fragment of the drug at X, L, or either of the two ends. For example, [ka] In this case, R1 is Re and R g from either end adjacent to L, R f and R h can bind to a functional fragment of the agent from the end adjacent to L, R f and R h from either end adjacent to L, R e and R g can bind to a functional fragment of the agent from the end adjacent to L, R

[0064] In some embodiments, R1 is, independently at each occurrence, unsubstituted or substituted -(alk) a -S-(alk) b -, -(alk) a -O-(alk) b -, -(alk) 655a -NR A -(alk) b -, -(alk) a -C(O)-(alk) b -, -(alk) a -C(S)-(alk) b -, -(alk) a -S(O)-(alk) b -, -(alk) a -S(O)2-(alk) b -, -(alk) a -OC(O)-(alk) b -, -(alk) a -C(O)O-(alk) b -, -(alk) a -OC(S)-(alk) b -, -(alk) a -C(S)O-(alk) b -, -(alk) a -C(O)NR A -(alk) b -, -(alk) a -C(S)NR A -(alk) b -, -(alk) a -S(O)2NR A -(alk) b -, -(alk)a -NR A C(O)-(alk) b -,-(alk) a -NR A C(S)-(alk) b -,-(alk) a -NR A S(O)2-(alk) b -,-(alk) a -NR A C(O)O-(alk) b -,-(alk) a -NR A C(S)O-(alk) b -,-(alk) a -OC(O)NR A -(alk) b -,-(alk) a -OC(S)NR A -(alk) b -,-(alk) a -NR A C(O)NR B -(alk) b -,-(alk) a -NR A C(S)NR B -(alk) b -and-(alk) a -NR A S(O)2NR B -(alk) b -mosquito Selected from the following groups, a and b are independently selected from the bases 0, 1, 2, 3, and 4. alk is independently C 1-5 Alkilen, C 1-5 Alkenylene and C 1-5 A selection is made from the group consisting of alkynylenes, each of which is H, halogen, -OH, NH2, CF3, C 1-5 Alkyl, -CH2(NH2), -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, -SH, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, -OC 1-5 Alkyl, -SC 1-5 Alkyl, -NH-C1-5 Alkyl and -N(C) 1~5 From the group consisting of alkyl)2, one to three substituents can be independently selected and optionally replaced with C 1~5 Alkyl groups include halogens, -OH, and -NH. 2、 C 1-4 One to three substituents selected from the group consisting of alkyl, CF3, -CH2(NH2), -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, -SH, -SCH3, imidazolyl, pyrazolyl, methylimidazolyl, and methylpyrazolyl may be independently and arbitrarily substituted; R A and R B In each appearance, hydrogen and C appear independently. 1-3 Alkyl, C 3-6 Selected from the group consisting of cycloalkyls, 5-10 membered heterocycles, aryls, and 5-10 membered heteroaryls, alkyls, cycloalkyls, heterocycles, aryls, and heteroaryls are halogens, C 1-3 Alkyl, OH, NH2, NH-C 1-3 Alkyl, N(C 1-3 Alkyl)2, CF3, C 1-6 One to three substituents selected from the group consisting of alkyl, -CH2(NH2), -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, -SH, -SCH3, imidazolyl, pyrazolyl, methylimidazolyl, and methylpyrazolyl, each of which can be independently and arbitrarily substituted. In each occurrence, R3 is independently selected from the following group: [ka] ; R in each appearance a , R b , R c , R d , R e , R f , R g and R hThese are independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 member heteroaryls, optionally substituted aryls, and optionally substituted 5-10 member heteroaryls, R a , R b , R c , R d , R e , R f , R g and R h The optional substituents are, independently, halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、 These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0065] In some embodiments, in each occurrence, R3 is independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, The optional substituents for R3 are halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents independently selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、 These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0066] In each occurrence, R4 is independently selected from the following group: [ka]

[0067] In some embodiments, in each occurrence, R4 is independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, The optional substituents of R4 are, independently, halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、 These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0068] In each occurrence, R5 is independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, The optional substituents of R5 are, independently, halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、 These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0069] In each occurrence, R6 is independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls. Selected from the following groups, The optional substituents of R6 are, independently, halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、 These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0070] In each occurrence, L is independently selected from the following group: [ka] , D1, D2, D3, D4, D5, and D6 are independently selected in each occurrence from the group consisting of N, C, O, or S, except D 1-6 If it is N, the corresponding position is trivalent, D 1-6If is O or S, the corresponding position is divalent. The linker can bind to either the CLM or the functional fragment of the drug at either of its two ends. For example, [ka] In this configuration, L can be bonded to CLM either from the end adjacent to nitrogen or from the other end.

[0071] In each occurrence, B1, B2, B3, and B4 are independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, The optional substituents of B1, B2, B3 and B4 are independently halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0072] In some embodiments, L is independently unsubstituted or substituted in each occurrence: -(alk)aS-(alk)b-, -(alk)aO-(alk)b-, -(alk)a-NRA-(alk)b-, -(alk)aC(O)-(alk)b-, -(alk)aC(S)-(alk)b-, -(alk)aS(O)-(alk)b-, -(alk)aS(O)2-(alk)b-, -(alk)a-OC(O)-(alk)b-, -(alk)aC(O)O-(alk)b-, -(alk)a-OC(S)-(alk)b-, -(alk)aC(S)O-(alk)b-, -(alk)aC(O)NR Selected from the group consisting of A-(alk)b-, -(alk)aC(S)NRA-(alk)b-, -(alk)aS(O)2NRA-(alk)b-, -(alk)a-NRAC(O)-(alk)b-, -(alk)a-NRAC(S)-(alk)b-, -(alk)a-NRAS(O)2-(alk)b-, -(alk)a-NRAC(O)O-(alk)b-, -(alk)a-NRAC(S)O-(alk)b-, -(alk)a-OC(O)NRA-(alk)b-, -(alk)a-OC(S)NRA-(alk)b-, -(alk)a-NRAC(O)NRB-(alk)b-, -(alk)a-NRAC(S)NRB-(alk)b-, and -(alk)a-NRAS(O)2NRB-(alk)b-, a and b are independently selected from the bases 0, 1, 2, 3, and 4. alk is independent of C 1-5 Alkilen, C 1-5 Alkenylene and C 1-5 Selected from the group consisting of alkynylenes, each independently comprises H, halogen, -OH, NH2, CF3, and C. 1-5Alkyl, -CH2(NH2), -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, -SH, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, -OC 1-5 Alkyl, -SC 1-5 Alkyl, -NH-C 1-5 Alkyl and -N(C 1~5 It is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl)2, C 1~5 Alkyl groups can independently be halogens, -OH, and -NH. 2、 C 1-4 The molecule is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, CF3, -CH2(NH2), -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, -SH, -SCH3, imidazolyl, pyrazolyl, methylimidazolyl, and methylpyrazolyl.

[0073] R C and R D In each appearance, hydrogen and C appear independently. 1-3 Alkyl, C 3-6 Selected from the group consisting of cycloalkyls, 5-10 membered heterocycles, aryls, and 5-10 membered heteroaryls, each alkyl, cycloalkyl, heterocycle, aryl, and heteroaryl can be independently of halogens, C 1-3 Alkyl, OH, NH2, NH-C 1-3 Alkyl, N(C 1-3 Alkyl)2, CF3, C 1-6 The molecule is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, -CH2(NH2), -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, -SH, -SCH3, imidazolyl, pyrazolyl, methylimidazolyl, and methylpyrazolyl.

[0074] In each occurrence, X is independently selected from the following group: [ka] [ka] [ka] [ka] [ka]

[0075] A, B, C, and D are, in each occurrence, independently H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinil, C is optionally substituted. 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, Any optional substituent for A, B, C, and D can be a halogen, OH, or NH, independently. 2、 CH 3、 CF 3、 -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH 3、 SH, -S-CH3, and C optionally substituted 1-3 Alkyl, optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、 These are one or two substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3.

[0076] A1, A2, A3, A4, A5, and A6 appear independently in each instance as H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, The optional substituents of A1, A2, A3, A4, A5, and A6 are independently halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, and optionally substituted C 1-3 Alkyl and optionally substituted C 3-6 These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. C 1-3 Alkyl and C 3-6 The optional substituents on the cycloalkyl group are halogen, OH, NH2, and CH. 3、1 to 2 substituents independently selected from the group consisting of CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH and -S-CH3.

[0077] This disclosure contemplates using all combinations of various substituents. Thus, any combination of the above-mentioned substituents included in the structural formulas of Compounds 1-1 to 1-5 can be used.

[0078] In some embodiments, ARCS is from Compound 1-6 to Compound 1-11 of the narrow general structure [Chemical formula] or selected from the group consisting of its pharmaceutically acceptable salts.

[0079] In some embodiments, ARCS [Chemical formula] may have the structure of or its pharmaceutically acceptable salt, L is a linker selected from the group consisting of [Chemical formula] and either end can be connected to CLM, R1 is selected from the group consisting of [Chemical formula] and CLM is selected from the group consisting of [Chemical formula] The compounds included in Formula 1-50 include, but are not limited to, Compounds 1-101, 1-102, 1-103, 1-104, 1-105, 1-106, 1-107, 1-108, 1-109, 1-110, 1-111, 1-112 in Table 1 , including 1-113, 1-114, 1-115, 1-116, 1-117, 1-118, 1-119, 1-120, 1-121, 1-122, 1-123, 1-124, 1-125, 1-126, 1-127, 1-128, 1-129, 1-130, 1-137, 1-138, 1-145, 1-146, 1-153, 1-154, 1-161, 1-162, 1-169, 1-170, 1-171 and 1-172.

[0080] In some embodiments, ARCS is [ka] It may have the structure or a pharmaceutically acceptable salt thereof, and the linker of L is [ka] And R2 is, [ka] Selected from the group consisting of, CLM is [ka] A compound is selected from the group consisting of the following. Compounds included by formula 1-51 are not limited to, but include, compounds 1-131, 1-132, 1-133, 1-134, 1-135, 1-136, 1-139, 1-140, 1-141, 1-142, 1-143, 1-144, 1-147, 1-148, 1-149, 1-150, 1-151, 1-152, 1-155, 1-156, 1-157, 1-158, 1-159, 1-160, 1-163, 1-164, 1-165, 1-166, 1-167, and 1-168 in Table 1.

[0081] An exemplary ARCS includes any compound selected from the group consisting of compounds 1-101 to 1-172. Table 1 Non-limiting examples of ARCS compounds [ka] [Chemical] [Chemical] [Chemical] [Chemical] [Chemical] [Chemical] [Chemical] [Chemical] [Chemical] [Chemical] or a pharmaceutically acceptable salt thereof.

[0082] E. Pharmaceutical Compositions The ARCS of the present disclosure may be administered to a subject using any convenient means that can produce the desired result. Thus, the ARCS of the present disclosure can be incorporated into various formulations for therapeutic administration. More specifically, the ARCS of the present disclosure may be formulated into a pharmaceutical composition by combination with a suitable pharmaceutically acceptable carrier or diluent, and may be formulated in solid, semi-solid, liquid, or gaseous forms such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, and aerosols.

[0083] As used herein, the term “pharmaceutical composition” means a composition comprising the ARCS described herein and at least one pharmaceutically acceptable carrier, e.g., any carrier commonly used in the pharmaceutical industry. The term “pharmaceutically acceptable” is used herein to mean a compound, material, composition and / or dosage form that is within the bounds of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0084] The administration of pharmaceutical compositions can be achieved by various methods, including oral, rectal, parenteral, intraperitoneal, intradermal, transdermal, and intratracheal administration. In pharmaceutical dosage forms, pharmaceutical compositions may be administered alone or in combination with other pharmaceutically active compounds.

[0085] The amount of ARCS in the pharmaceutical composition may be based on weight, moles, or volume. In some embodiments, the pharmaceutical composition contains at least 0.0001% ARCS. In some embodiments, the pharmaceutical composition contains at least 0.1% ARCS. In some embodiments, the pharmaceutical composition contains at least 0.5% ARCS. In some embodiments, the pharmaceutical composition contains at least 1% of the ARCS compound. In some embodiments, the pharmaceutical composition contains at least 2% ARCS. In some embodiments, the pharmaceutical composition contains at least 3% ARCS. In some embodiments, the pharmaceutical composition contains at least 4% ARCS. In some embodiments, the pharmaceutical composition contains at least 5% ARCS. In some embodiments, the pharmaceutical composition contains at least 10% ARCS. In some embodiments, the pharmaceutical composition contains 0.05% to 90% ARCS. In some embodiments, the pharmaceutical composition contains 0.1% to 85% ARCS. In some embodiments, the pharmaceutical composition contains 0.5% to 80% ARCS. In some embodiments, the pharmaceutical composition contains 1% to 75% ARCS. In some embodiments, the pharmaceutical composition contains 2% to 70% ARCS. In some embodiments, the pharmaceutical composition contains 3% to 65% ARCS. In some embodiments, the pharmaceutical composition contains 4% to 60% ARCS. In some embodiments, the pharmaceutical composition contains 5% to 50% ARCS.

[0086] It will also be understood that certain ARCSs may exist in free form for therapeutic purposes or, where appropriate, as pharmaceutically acceptable derivatives thereof. According to this disclosure, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or prodrugs or other adducts or derivatives of ARCSs, and ARCSs may be included in compositions that, at the time of administration to a patient in need, directly or indirectly provide the compound or its metabolites or residues as described herein.

[0087] As described above, the pharmaceutical compositions of this disclosure may contain pharmaceutically acceptable excipients, which, when used herein, include any and all solvents, diluents, or other liquid vehicles, dispersants or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, antioxidants, solid binders, lubricants, etc., suitable for the specific dosage form of the choice.

[0088] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, magnesium talc, calcium or zinc stearate or stearic acid) or solvent encapsulating material involved in transporting or carrying the compound of interest from one organ or part of the body to another. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and is not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, methylcellulose, ethylcellulose, microcrystalline cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricants such as magnesium stearate, sodium lauryl sulfate, and talc; (8) excipients such as cocoa butter and suppository wax; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; and (10) glycols such as propylene glycol. (11) Polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) Esters such as ethyl oleate and ethyl laurate; (13) Agar; (14) Buffers such as magnesium hydroxide and aluminum hydroxide; (15) Arginic acid; (16) Pyrogen-free water; (17) Isotonic saline; (18) Ringer's solution; (19) Ethyl alcohol; (20) pH buffer solution; (21) Polyesters, polycarbonates, and / or polyanhydrides; (22) Bulking agents such as polypeptides and amino acids; (23) Serum components such as serum albumin, HDL, and LDL; (24) C2-C12 alcohols such as ethanol; and (25) Other non-toxic compatible substances used in pharmaceutical formulations. Wetting agents, colorants, release agents, coating agents, sweeteners, flavoring agents, fragrances, preservatives, and antioxidants may also be present in the formulation.In this specification, terms such as “excipient,” “carrier,” and “pharmaceutically acceptable carrier” are used interchangeably.

[0089] Useful pharmaceutical carriers for the preparation of the compositions described herein may be solid, liquid, or gas. Suitable pharmaceutical carriers and their formulations are described in Remington's Pharmaceutical Sciences by E.W. Martin. Such compositions, in any case, contain an effective amount of ARCS together with a suitable carrier to prepare a suitable dosage form for appropriate administration to the recipient.

[0090] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to ARCS, liquid dosage forms include, for example, water or other solvents, solubilizers, and emulsifiers. For example, the oral composition may include inactivating diluents commonly used in the art, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan and mixtures thereof. In addition to inactivating diluents, the oral composition may also include adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0091] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, ARCS comprises at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, such as carboxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) hydrating agents such as glycerol; d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarders such as paraffin; f) absorption enhancers such as quaternary ammonium compounds; g) wetting agents, such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite clay; and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, and sodium lauryl sulfate, as well as mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include a buffer.

[0092] Similar solid compositions can also be used as fillers in soft and rigid gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol. Solid dosage forms of tablets, sugars, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other coatings well known in pharmaceutical formulation technology. These solid dosage forms may optionally contain opacifiers and may be compositions that release only the active ingredient, or optionally release it preferentially, in a delayed manner, in a specific part of the intestinal tract. Examples of embedding compositions that can be used include polymers and waxes. Similar solid compositions can also be used as fillers in soft and rigid gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0093] ARCS may also be in the form of microcapsules having one or more excipients, as described above. Solid dosage forms of tablets, sugars, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and controlled-release coatings, as well as other coatings known in pharmaceutical formulation technology. In these solid dosage forms, ARCS may be mixed with at least one inert diluent, such as sucrose, lactose, and starch. These dosage forms may also contain additional substances other than the inert diluent, as is common practice, such as tableting lubricants and magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also contain buffers. These solid dosage forms may optionally contain opacifiers and may be compositions that release only the active ingredient, or preferentially, selectively, and delayedly, in a specific portion of the intestinal tract. Examples of embedding compositions that may be used include polymers and waxes.

[0094] Formulations suitable for parenteral administration conveniently, and preferably, contain a sterile aqueous preparation of the drug that is isotonic with the recipient's blood. Suitable excipient solutions include phosphate-buffered saline, saline, water, Ringer's lactate solution, or dextrose (5% in water). Such formulations can be conveniently prepared by mixing the drug with water to produce a solution or suspension. The formulation is filled into sterile containers and sealed to prevent bacterial contamination. The sterile material is preferably used under sterile manufacturing conditions to avoid the need for final sterilization. Such formulations may optionally contain one or more additional components, including preservatives such as methyl hydroxybenzoate, chlorocresol, metaczol, phenol, and benzalkonium chloride. Such substances are of particular value when the formulation is presented in multi-dose containers.

[0095] A buffer solution is also included to provide the appropriate pH value for the formulation. Suitable buffer materials include sodium phosphate and sodium acetate. Sodium chloride or glycerin can be used to make the formulation isotonic using blood.

[0096] If desired, the formulation can be filled into a container under an inert atmosphere such as nitrogen, and can be conveniently presented in single-dose or multi-dose forms, for example, in sealed ampoules.

[0097] Those skilled in the art will recognize that the amounts of various components of the composition of the Disclosure administered to a subject according to the method of the Disclosure depend on the factors described above.

[0098] Preparations for injection can be formulated according to known techniques, for example, by using appropriate dispersants or wetting agents and suspending agents to sterile aqueous or oily suspensions for injection. Sterile preparations for injection may also be sterile solutions, suspensions, or emulsions for injection in non-toxic, parenterally acceptable diluents or solvents, such as a solution in 1,3-butanediol. Acceptable vehicles and solvents that may be used include water, Ringer's solution, USP, and isotonic sodium chloride solution. Furthermore, sterile fixatives are conventionally used as solvents or suspension media. For this purpose, any non-irritating fixative, including synthetic monoglycerides or diglycerides, can be used. In addition, fatty acids such as oleic acid are used in the preparation of injections.

[0099] A non-limiting example of a tablet may contain an amount of active ingredient ranging from 10 mg to 100 mg, 70 mg to 95 mg of powdered lactose, 10 mg to 35 g of white corn starch, 1 mg to 8 mg of polyvinylpyrrolidone, 1 mg to 10 mg of sodium (Na) carboxymethyl starch (CMS), and 1 mg to 5 mg of magnesium stearate, with a tablet weight ranging from 200 mg to 3000 mg.

[0100] Examples of tablets in this disclosure are as follows: Ingredients: mg / tablet Active ingredient 100 Lactose powder 95 White corn starch 35 Polyvinylpyrrolidone 8 Sodium carboxymethyl starch 10 Magnesium stearate 2 Tablet weight 250

[0101] A non-limiting example of a capsule may contain an amount of active ingredient ranging from 10 mg to 100 mg, 50 mg to 75 mg of crystalline lactose, 10 mg to 35 mg of microcrystalline cellulose, 1 mg to 8 mg of talc, and 1 mg to 5 mg of magnesium stearate, with a capsule filling weight ranging from 100 mg to 3000 mg.

[0102] Examples of capsules in this disclosure are as follows: Ingredients mg / capsule Active ingredient 50 Crystalline lactose 60 Microcrystalline cellulose 34 Talc 5 Magnesium stearate 1 Capsule filling weight: 150

[0103] In the capsules shown above, the active ingredients have an appropriate particle size. Crystalline lactose and microcrystalline cellulose are homogeneously mixed and sieved, and then talc and magnesium stearate are mixed in. The final mixture is filled into appropriately sized hard gelatin capsules.

[0104] A non-limiting example of injection is an amount of active ingredient ranging from 0.05 mg to 5 mg, 10.0 μL to 20.0 μL of 1N HCl, 0.1 mg to 1 mg of acetic acid, 1 mg to 10 mg of sodium chloride, 1 mg to 10 mg of phenol, and enough 1N HCl to adjust the pH to 4 to 5. Contains NaOH and a sufficient amount of water.

[0105] Examples of the injectable solutions described herein are as follows: Component mg / solution Active substance 1.0mg 1 N HCl 20.0 μl Acetic acid 0.5 mg NaCl 8.0mg Phenol 10.0 mg 1N NaOH qsad pH5 H2O qsad 1mL

[0106] Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0107] To extend the effects of ARCS, it is often desirable to slow down the absorption of ARCS from subcutaneous or intramuscular injection. This can be achieved by using a low-water-soluble liquid suspension or amorphous material. The absorption rate of ARCS then depends on its dissolution rate, and then on its crystal size and crystalline form. Alternatively, delayed absorption of parenterally administered ARCS forms is achieved by dissolving or suspending ARCS in an oil vehicle. Depot formulations for injection are prepared by forming a microcapsule matrix of ARCS in a biodegradable polymer such as polylactide-polyglycolide. The rate of ARCS release can be controlled depending on the ratio of ARCS to polymer and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydrous). Depot injection formulations are also prepared by encapsulating ARCS in liposomes or microemulsions compatible with body tissues.

[0108] The composition for rectal or vaginal administration is preferably a suppository that can be prepared by mixing ARCS with a suitable non-irritating excipient or carrier such as cocoa butter or polyethylene glycol, or with a suppository wax that is solid at ambient temperature but liquid at body temperature and therefore melts in the rectum or vaginal cavity to release the active compound.

[0109] Typical suppository formulations contain ARCS or a pharmaceutically acceptable salt thereof, and when administered in this manner, for example, polymer glycol, gelatin, cocoa butter or other low-melting substances. It is active with binders and / or lubricants such as plant waxes or fats. Typical transdermal formulations include conventional aqueous or non-aqueous vehicles, such as creams, ointments, lotions or pastes, or in the form of medicinal plastics, patches or films.

[0110] Typical compositions for inhalation are in the form of solutions, suspensions, or emulsions, which can be administered in aerosol form using conventional sprays such as dichlorodifluoromethane or trichlorofluoromethane.

[0111] Depending on the route of administration, those skilled in the art can determine and adjust the effective dose of the small molecules disclosed herein for subjects such as human subjects accordingly.

[0112] Toxicity and therapeutic efficacy are, for example, LD 50 (50% lethal dose in the population) and ED 50 The effective dose for 50% of the population can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The dose-to-toxicity ratio is the therapeutic index, or LD50. 50 / ED 50 It can be expressed as a ratio. Compositions exhibiting a large therapeutic index are preferred.

[0113] The descriptions of pharmaceutical compositions provided herein primarily concern pharmaceutical compositions suitable for administration to humans, but those skilled in the art will understand that such compositions are generally suitable for administration to any other animals, such as non-human animals, such as non-human mammals. Targets to which the pharmaceutical compositions are intended include, but are not limited to, agricultural animals such as cattle, horses, chickens, and pigs; domestic animals such as cats and dogs; or research animals such as mice, rats, rabbits, dogs, and non-human primates, as well as non-human mammals.

[0114] II. How to use ARCS The ARCS described herein or compositions containing the ARCS described herein may be administered to treat any therapeutic disease that can be treated with its FCB or any therapeutic disease related to the biological targets of the ARCS, including but not limited to cancer, neurodegenerative diseases, autoimmune diseases, or aging, as appropriate. The formulations can be delivered to various body parts, such as the brain and central nervous system, eyes, ears, lungs, bones, heart, kidneys, liver, spleen, breasts, ovaries, colon, pancreas, muscles, gastrointestinal tract, mouth, and skin, to treat diseases associated with these body parts. The formulations may be administered by injection, orally, or topically, typically to mucosal surfaces (lungs, nasal cavity, oral cavity, buccal, sublingual, vagina, rectum) or eyes (intraocular or transocular).

[0115] In aspects of this disclosure, ARCS binds to a biological target. In some embodiments, the biological target includes, but is not limited to, kinases such as phosphoinositide 3-kinase (PI3K) and pseudokinase.

[0116] In some embodiments, ARCS can form covalent bonds with biological targets. In some embodiments, ARCS can form covalent bonds with biological targets from about 5% to 100% of the biological target. In some embodiments, ARCS can form covalent bonds with biological targets from about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the biological target.

[0117] In some embodiments, ARCS can form covalent bonds with PI3 kinase. In some embodiments, ARCS can form covalent bonds with PI3 kinase from about 5% to 100% of PI3 kinase. In some embodiments, ARCS can form covalent bonds with PI3 kinase from about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, and 5% of PI3 kinase. It can form a covalent bond with PI3 kinase at concentrations of 0%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

[0118] Protein kinases and pseudokinases Protein kinases and pseudokinases regulate signaling pathways for a wide range of biological processes in both normal and diseased states (Brognard, J. et al., Curr. Opin. Genet. Dev. 2011, 21, pp4-11; Cohen, P. et al., Nat. Cell Biol. 2002, 4, E127-130). In fact, most eukaryotic processes are controlled by kinases and / or pseudokinases. The term "pseudokinase" refers to proteins that have a kinase domain lacking catalytically relevant residues, including lysine in the VAIK motif or aspartic acid in the DFG motif. Examples of pseudokinases include HER3, STRAD, ILK, KSR1, and KSR2. Some pseudokinases are incapable of phosphoryl group transduction, while others possess the ability to transduce phosphoryl groups despite lacking catalytically relevant residues.

[0119] Kinases and pseudokinases can function by inhibiting and / or activating their protein partners through processes such as binding, covalent modification including phosphorylation, conformational regulation, cellular localization, and / or other processes. Protein kinases and pseudokinases can promote and / or antagonize a wide range of diseases, including but not limited to cancer, Alzheimer's disease, aging, diabetes, cardiovascular disease, CNS-related disorders, immunological disorders, allergies, hypertension, and Parkinson's disease. Protein kinases and pseudokinases are also commonly mutated in several diseases, including but not limited to cancer and Parkinson's disease.

[0120] Molecules targeting kinases and pseudokinases can act as anticancer agents. Unfortunately, common inhibitors often exhibit limited efficacy due to their short resonance times at the target. Therefore, there is a need to discover kinase inhibitors that, compared to previous kinase inhibitors, have improved resonance times at the target and simultaneously possess improved potential and efficacy.

[0121] In some embodiments, the ARCS of this disclosure may target kinases and pseudokinases, thereby inhibiting them. The FCB of the ARCS may be an inhibitor of the kinase and pseudokinase. The CLM of the ARCS may be covalently bound to the kinase and pseudokinase.

[0122] In some embodiments, the Disclosure provides a method for treating a disease comprising administering a therapeutically effective amount of the ARCS or ARCS-containing composition or a pharmaceutically acceptable salt thereof to a patient in need thereof, the patient having a disease partially or entirely caused by alterations in the modulation of a target kinase or pseudokinase, such as cancer, Alzheimer's disease, aging, diabetes, cardiovascular disease, CNS-related disease, immunological disease, allergy, hypertension, or Parkinson's disease. In some embodiments, the disease is related to PI3K or involves a mutation in PI3K. In some embodiments, the subject has cancer having a mutation in the PIK3CA gene.

[0123] Administration This disclosure provides a method comprising administering a composition comprising the ARCS described herein to a subject in need thereof. The ARCS composition comprising herein may be administered to a subject using any amount and any route of administration that is effective in preventing, treating or imaging a disease, disorder and / or condition. The exact amount required is determined by the subject. The appropriate treatment varies depending on the target individual, including the species, age and overall health condition, the severity of the disease, the specific composition, the mode of administration, and the mode of activity.

[0124] The compositions according to this disclosure are typically formulated in unit dosage forms for ease of administration and dose uniformity. However, it is understood that the total daily dose of the compositions according to this disclosure may be determined by the attending physician within the bounds of sound medical judgment. For any particular patient, a specific therapeutic effective dose, prophylactic effective dose, or appropriate imaging dose level depends on a variety of factors, including the disorder being treated and its severity, the activity of the specific compound used, the specific composition used, the patient's age, weight, overall health, sex, and diet, the timing of administration, route of administration, and elimination rate of the specific compound used, the duration of treatment, any drugs used in combination with or incidentally with the specific compound used, and factors well known in the medical field.

[0125] In some embodiments, the compositions according to the present disclosure may be administered once or more times daily at a dose level sufficient to deliver approximately 0.0001 mg / kg to approximately 100 mg / kg to approximately 0.001 mg / kg to approximately 0.05 mg / kg to approximately 0.005 mg / kg to approximately 0.05 mg / kg to approximately 0.001 mg / kg to approximately 0.005 mg / kg to approximately 0.05 mg / kg to approximately 0.5 mg / kg to approximately 0.01 mg / kg to approximately 50 mg / kg to approximately 0.1 mg / kg to approximately 40 mg / kg to approximately 0.5 mg / kg to approximately 30 mg / kg to approximately 0.01 mg / kg to approximately 10 mg / kg to approximately 0.1 mg / kg to approximately 10 mg / kg or approximately 1 mg / kg to approximately 25 mg / kg of the subject's body weight per day to obtain the desired therapeutic, diagnostic, prophylactic, or imaging effect. The desired dose can be delivered three times a day, twice a day, once a day, every other day, every three days, weekly, every two weeks, every three weeks, or every four weeks. In some embodiments, the desired dose may be delivered using multiple doses (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more doses). When multiple doses are employed, a divided dosing regimen, such as those described herein, may be used.

[0126] As used herein, “divided dose” means dividing a single unit dose or total daily dose into two or more doses, such as two or more single unit doses. As used herein, “single unit dose” is the dose of any therapeutic agent administered in a single dose event, i.e., a single administration event. As used herein, “total daily dose” is the amount administered or prescribed over a 24-hour period. The total daily dose may be administered as a single unit dose.

[0127] III. Kits and Equipment This disclosure provides various kits for conveniently and / or effectively carrying out the methods of this disclosure. Typically, the kits contain sufficient quantities and / or a number of components, allowing the user to perform multiple treatments and / or experiments on a target.

[0128] In one embodiment, the Disclosure provides a kit for inhibiting tumor cell growth in vitro or in vivo, the kit comprising the ARCS of the Disclosure or a combination of the ARCS of the Disclosure in any other active agent in optional combination.

[0129] The kit may further include packaging and / or instructions for forming the formulation composition, and / or a delivery agent. The delivery agent may include saline, a buffer solution, or any delivery agent disclosed herein. The amounts of each component may be varied to allow for consistent, reproducible higher concentrations of saline or simple buffer formulations. The components may also be varied to increase the stability of ARCS in the buffer over a period of time and / or under various conditions.

[0130] This disclosure provides devices that can incorporate the ARCS of this disclosure. These devices It is contained in a stable formulation that can be immediately delivered to a target that needs it, such as a human patient. In some embodiments, the target has cancer.

[0131] Non-limiting examples of the apparatus include pumps, catheters, needles, transdermal patches, pressurized olfactory delivery devices, ionophoresis devices, and multilayer microfluidic devices. The apparatus may be used to deliver the conjugates and / or particles of the Disclosure in single, multiple, or divided dosing regimens. The apparatus may be used to deliver the conjugates and / or particles of the Disclosure into biological tissue, intradermally, subcutaneously, or intramuscularly.

[0132] A. assay Covalent binding of ARCS to biological targets includes enzyme-linked immunosorbent assay (ELISA), gel assay, antibody array, Western blot, affinity ELISA, ELISPOT, immunochemistry (e.g., IHC), in situ hybridization (ISH), flow cytometry, immunocytology, surface plasmon resonance analysis, kinetic exclusion assay, liquid chromatography-mass spectrometry (LCMS), tandem mass spectrometry (MS / MS), high-performance liquid chromatography (HPLC), BCA assay, immunoelectrophoresis, SDS-PAGE, protein immunoprecipitation, and / or PCR.

[0133] As used herein, the term “assay” refers to a set of activities related to the reported results, which may include, but are not limited to, cell seeding, preparation of test material, infection, lysis, analysis, and calculation of results.

[0134] In some embodiments, the assay surface on the substrate is sterile and suitable for culturing cells under conditions representing culture conditions during the large-scale (e.g., industrial-scale) production of biological products. In some embodiments, the exterior of the substrate includes wells, depressions, borders, etc., at locations corresponding to the assay surface. In some embodiments, the wells, depressions, borders, or similar structures hold a fluid, such as cell culture medium, on the assay surface.

[0135] In some embodiments, the substrate includes a microarray plate, a biochip, etc., enabling high-throughput, automated testing of a series of test agents, conditions, and / or combinations thereof related to the production of biological products by cultured cells. For example, the substrate may include a two-dimensional microarray plate or biochip having m-columns and n-row assay surfaces (e.g., present in wells), enabling testing of m x n combinations of test agents and / or conditions (e.g., on a 24-well, 96-well, or 384-well microarray plate). The microarray substrate is preferably designed so that all necessary positive and negative controls can be performed in parallel with the testing of agents and / or conditions.

[0136] B. Screening Methods Generally, the synthesis of therapeutic conjugates that form covalent bonds with biological targets involves multiple synthesis and purification steps. When using these synthesis and purification steps, generating libraries of therapeutic conjugates or developing structure-activity relationships (SARs) for screening purposes can be challenging. There remains a need for methods and systems for automatically generating libraries of therapeutic conjugates using organic synthesis methods. To discover therapeutic conjugate drug leads, libraries of therapeutic conjugates against protein receptors must be screened to identify molecules that specifically bind to receptors in the cellular environment, and this binding is covalent and irreversible. Identifying whether a therapeutic conjugate covalently binds to a specific target within a cell is difficult to prove.

[0137] Current methods in the art for screening therapeutic conjugates for their covalent binding to biological targets include tandem mass spectrometry approaches. Tandem MS, or MS / MS, is a method for decomposing selected ions into fragment ions. Once a sample is ionized to produce a mixture of ions, a precursor ion with a specific mass-to-charge ratio (m / z) is selected (MS1) and then fragmented (MS2) to produce a product ion for detection. Information about the chemical structure of the selected ion can be determined from the fragments.

[0138] However, there are challenges associated with mass spectrometry. Mass spectrometry is primarily limited to fragment screening rather than drug-like molecule screening. Mass spectrometry analysis of therapeutic conjugates often results in unresolved analyses because it is difficult to identify some of the fragments. This approach involves a multi-step process with long processing times and manual analysis. MS / MS is difficult to interpret because it requires defragmenting the complex, larger molecules, and manually reassembling them by each peptide. Another drawback of using mass spectrometry is that detection is proportional to ionization rather than abundance, making the technique somewhat quantitative.

[0139] Therefore, there are challenges associated with the synthesis and screening of therapeutic conjugates. Firstly, it is difficult to generate targeted covalent inhibitors that can access both cysteine ​​and non-cysteine ​​amino acids. Secondly, it is difficult to distinguish false-positive covalent inhibitors from actual positives.

[0140] To address the aforementioned problems, the inventors combined a combinatorial synthesis approach with a reliable screening approach to identify potential therapeutic conjugates. This disclosure provides a high-throughput complex approach for synthesizing therapeutic conjugates, rapidly tracking covalent binding, analyzing large libraries for duration of action, and directly quantifying intracellular covalent target binding.

[0141] By using the MS / MS method, thousands of molecules can be screened per day with picomolar sensitivity, compared to tens of molecules with nanomolar sensitivity. This disclosure is applicable to any drug molecule, is not limited to fragments, and provides quantitative results with 95–99% reproducibility.

[0142] In some embodiments, the method for screening the ARCS library is as follows: To generate an ARCS library in the composition, The process involves bringing the library into contact with target cells, Lysizing target cells to produce solubilizes, Labeling the solubilized substance, and This includes detecting the covalent binding of ARCS to biological targets on target cells.

[0143] Examples of human cell lines useful as target cells in the methods provided herein include, but are not limited to, 293T (embryonic kidney), 786-0 (renal), A498 (kidney), A549 (alveolar basal epithelium), ACHN (renal), BT-549 (breast), BxPC-3 (pancreas), CAKI-1 (renal), Capan-1 (pancreas), CCRF-CEM (leukemia), COLO205 (colon), DLD-1 (colon). intestine), DMS114 (small cell lung), DU145 (prostate), EKVX (non-small cell lung), HCC-2998 (colon), HCT-15 (colon), HCT-116 (colon), HT29 (colon), HT-10 80 (fibrosarcoma), HEK293 (embryonic kidney), HeLa (cervical cancer), HepG2 (hepatocellular carcinoma), HL-60 (TB) (leukemia), HOP-62 (non-small cell lung), HOP-92 (non-small cell lung), HS 578T (breast), HT-29 (colon adenocarcinoma), IGR-OV1 (ovary), IMR32 (neuroblastoma), Jurkat (T lymphocyte), K-562 (leukemia), KM12 (colon), KM20L2 (colon), LAN5 (neuroblastoma), LNCap.FGC (white prostate adenocarcinoma), LOX IMVI (melanoma), LXFL529 (non-small cell lung), M14 (melanoma), M19-MEL (melanoma), MALME-3M (melanoma), MCFlOA (mammary epithelium), MCF7 (mammary gland), MDA-MB-453 (mammary epithelium), MDA-MB-468 (breast), MDA-MB-231 (breast), MDA-N (breast), MOLT-4 (leukemia), NCI / ADR-RES (ovary), NCI-H226 (non-small cell lung), NCI-H23 (non- Small cell lung, NCI-H322M (non-small cell lung), NCI-H460 (non-small cell lung), NCI-H522 (non-small cell lung), OVCAR-3 (ovary), OVCAR-4 (ovary), OVCAR-5 (ovary), OVCAR-8 (ovary), P388 (leukemia), P388 / ADR (leukemia), PC-3 (prostate), PERC6 (registered trademark) (E1 transformed embryonic retina), RPMI-7951 (melanoma), RPMI-8226 (leukemia), RXF 393 (renal), RXF-631 (renal), Saos-2 (bone), SF-268 (CNS), SF-295 (CNS), SF-539 (CNS), SHP-77 (small cell lung), SH-SY5Y (neuroblast) cystoma), SK-BR3 (breast), SK-MEL-2 (melanoma), SK-MEL-5 (melanoma), SK-MEL-28 (melanoma), SK-OV-3 (ovary), SN12K1 (renal), SN12C (renal), S Examples include NB-19 (CNS), SNB-75 (CNS), SNB-78 (CNS), SR (leukemia), SW-620 (colon), T-47D (breast), THP-1 (monocyte-derived macrophage), TK-10 (renal), U87 (glioblastoma), U293 (kidney), U251 (CNS), UACC-257 (melanoma), UACC-62 (melanoma), UO-31 (kidney), W138 (lung), and XF498 (CNS).

[0144] Examples of rodent cell lines useful for the methods provided herein include, but are not limited to, baby hamster kidney (BHK) cells (e.g., BHK21 cells, BHK TK- cells), mouse Sertoli (TM4) cells, buffalo rat liver (BRL 3A) cells, mouse mammary tumor (MMT) cells, rat hepatoma (HTC) cells, mouse myeloma (NS0) cells, mouse hybridoma (Sp2 / 0) cells, mouse thymoma (EL4) cells, Chinese hamster ovary (CHO) cells and CHO cell derivatives, mouse embryonic (NIH / 3T3, 3T3Ll) cells, rat cardiomyocyte (H9c2) cells, mouse myoblast (C2C12) cells, and mouse kidney (miMCD-3) cells.

[0145] Examples of non-human primate cell lines useful for the methods provided herein include, but are not limited to, monkey kidney (CVI-76) cells, African green monkey kidney (VERO-76) cells, green monkey fibroblast (Cos-1) cells, and monkey kidney (CVI) cells transformed with SV40 (Cos-7). Additional mammalian cell lines are known to those skilled in the art and are cataloged in the American Type Culture Collection catalog (ATCC®, Mamasus, Virginia).

[0146] In some embodiments, cells are lysed using chemical and / or mechanical lysis. In some embodiments, chemical lysis involves a lysis buffer containing a protease inhibitor, phosphate-buffered saline, and Triton X100. In some embodiments, cells may be frozen after being added to the lysis buffer at -80°C for about 30 minutes to about 72 hours. Alternatively, the cell lysates may be stored at a temperature range of 2–8°C or at room temperature. In some embodiments, cells are centrifuged and the cell lysates are collected. In some embodiments, this is carried out by rotating the cells in a centrifuge at 3,750 RPM at room temperature for 10 minutes.

[0147] The methods described herein are not limited to, but may include, cell plates such as 24-well plates, 48-well plates, 96-well plates, or 384-well plates, individual cell culture plates, or flasks such as T-flasks or shaking flasks. This can be carried out by utilizing any of the wide range of cell assay formats, including [specific format / method].

[0148] The covalent bonds of ARCS can be detected by assays such as gel assays, NanoBRET assays, Western blots, ELISAs, or microarrays, but are not limited to these. For example, gel assays, including microfluidic or capillary techniques, can separate proteins by size.

[0149] In some embodiments, covalent bonding is detected by gel assay. “Gel assay” is defined as an assay in which cells or cell solubilates are first treated with ARCS at doses of 1 picomole–1 mmol for a period of 2 minutes to 120 hours using techniques known to those skilled in the art, including but not limited to common cell culture techniques. The cells are then lysed using techniques known to those skilled in the art, including but not limited to sonication or buffer lysis. The resulting solubilizes, also described as unpurified cell solubilates, can be further prepared and used using techniques known to those skilled in the art, including but not limited to centrifugation, to produce purified solubilizes. Purified or unpurified solubilizes are likely to contain proteins covalently bonded to the molecule of interest. “Coupling reagents” and labeling molecules are added to the purified or unpurified solubilizes, and the ARCS in the reaction mixture is covalently labeled with the labeling molecules via a copper-free or copper-driven click reaction. The compounds bonded to the labeling molecules are then added, enabling reliable stoichiometric separation and reliable covalent drug tracking. Next, the sample is subjected to a Western blotting method well known to those skilled in the art. The amount of covalent bonding can then be tracked based on the shift of the drug-treated band compared to the untreated band. The band can be quantified using densitometry, and the quantitative amount of covalent label can be determined using the relative abundance of the band.

[0150] Generally, covalent binding of high molecular weight proteins / mass to ARCS leads to a shift in the target-ARCS-high molecular weight protein-mass complex in the gel. If azide-bonded molecules, which are bound to high molecular weight proteins or any type of molecule, are directly linked to alkynes on the ARCS, the shift still occurs, making it possible to detect the covalent binding of ARCS to targets.

[0151] In some embodiments, covalent bonding is detected by a gel-only shift assay. A "gel-only shift assay" is defined as an assay in which a protein is expressed in any cell type (via transfection or infection) and linked to a tagging domain. This tagging domain includes a fluorescent protein or a linker protein. Fluorescent proteins include GFP, RFP, etc. Linker proteins include HALO, SNAP-, CLIP-, ACP-, and MCP- tags. After transfection or infection, cells are then treated with a therapeutic conjugate of the Disclosure, which contains an alkyne that can potentially covalently bond. The cells are then lysed. In the case of protein expression with a linker protein, a "coupling reagent" is then added to covalently bond a fluorescent dye to the target protein. The solubilizer is then run on a gel, and the target protein can be visualized via in-gel fluorescence without the need for Western blot transfer. The amount of covalent bonding can be tracked based on the shift of the drug-treated band compared to the untreated band. The band can be quantified using densitometry, and the relative abundance of the band can be used to determine the quantitative amount of the covalent label.

[0152] The tagging domain can be any domain that enables the labeling of the target. In some embodiments, the tagging domain includes a label. This label is applied by the antibody. The domain itself may contain a recognized epitope or a detectable light or radioactive label. In some embodiments, the label is a fluorescent marker, e.g., FITC, phycobiliprotein, e.g., R- or B-phycoerythrin, allophycocyanin, AlexaFluor dye, Cy3, Cy5, Cy7, luminescent marker, 125 I or 32 The group consists of a radioactive label such as P, an enzyme such as horseradish peroxidase or alkaline phosphatase, or an alkaline phosphatase, such as alkaline shrimp phosphatase.

[0153] In some embodiments, as used herein, “fluorescent protein” includes, but is not limited to, Aequorea victoria green fluorescent protein (GFP), red fluorescent protein (RFP), structural variants of GFP (i.e., cyclic displacement, monomeric form), folding variants of GFP (i.e., more soluble form, superfolder form), spectral variants of GFP (i.e., YFP, CFP), and GFP-like fluorescent proteins (i.e., Dsked). The term “GFP-like fluorescent protein” is used to refer to members of Anthozoa fluorescent proteins that share the 11-beta-chain barrel structure of GFP as well as its structure, folding, and spectral variants. “GFP-like non-fluorescent protein” and “GFP-like chromogenic protein” (or simply “chromogenic protein” or “pigment protein”) are used to refer to anthozoan and hydrozoan chromogenic proteins that share the 11-beta-chain barrel structure of GFP as well as its structure, folding, and spectral variants.

[0154] In some embodiments, covalent binding is detected by a Western blot-based shift assay. A "Western blot-based shift assay" is defined as an assay in which a sample is performed via a Western blotting method well known to those skilled in the art. Covalent proteins then shift compared to non-covalent bands, allowing the amount of covalent binding to be tracked based on the target shift. The bands can be quantified using densitometry, and the relative abundance of the bands can be used to determine the quantitative amount of covalent labeling.

[0155] In some embodiments, covalent binding is detected by an ELISA assay. In some embodiments, covalent binding is detected by ELISA assay 1. "ELISA assay 1" is defined as an assay in which a solubilization solution containing a biotin-labeled drug is immobilized on a solid support via hybridization with monomeric or tetrameric streptavidin or streptavidin variants or molecules that bind to biotin. After immobilization of the drug, a detection antibody is added to detect the drug target of interest. The detection antibody may be covalently bound to an enzyme, or it may be detected itself by a secondary antibody linked to an enzyme or fluorescent label, through a bioconjugated reaction. Between each step, the plate is washed with solution to remove any nonspecifically bound proteins or antibodies. After the final washing step, the plate is developed by adding an enzyme substrate to generate a visible signal indicating the amount of covalent drug binding to the target of interest. The amount of covalent binding can be tracked based on the amount of signal.

[0156] In some embodiments, covalent binding is detected by ELISA assay 2. "ELISA assay 2" is defined as an assay immobilized on a solid support via hybridization with an antibody that binds to the target of interest. After immobilizing the target of interest, a detection antibody, monomeric / tetrameric streptavidin, or streptavidin variant is added to detect the drug bound to the target of interest. The detection antibody, monomeric / tetrameric streptavidin, or streptavidin variant may be covalently bound to an enzyme or fluorescent label, or may be detected by a secondary antibody linked to the enzyme or fluorescent label by a bioconjugated reaction. Between each step, the plate is washed with solution to remove any residues. Removes any specifically bound proteins or antibodies. After the final washing step, the plate is developed by adding an enzyme substrate to generate a visible signal, or by directly measuring the fluorescence signal to indicate the amount of covalent drug binding to the target of interest. The amount of covalent binding can be tracked based on the amount of signal.

[0157] In some embodiments, covalent binding is detected by an antibody array. An “antibody array” is defined as a system in which individual or multiple antibodies are bound to a solid support, enabling the detection of target proteins bound to those antibodies and molecules bound to those target proteins. An antibody microarray consists of a series of individual dots or wells in which a specific antibody is hybridized to each dot or well (as described in U.S. Patent No. 20120231963A1, the contents of which are incorporated herein by reference in their entirety). A bound purified or unpurified solubilized solution is added to the “antibody microarray” to enable the separation of different proteins, the localization of specific proteins to antibody-binding partners, or the washing of additional proteins. Labeled target molecules are detected in each microarray dot or well to determine the amount of covalent binding of the target molecule to a specific protein or multiple proteins by detecting the presence of biotin-labeled molecules and revealing the labeling level. The labeling level at each dot indicates the amount of covalent labeling of the specific protein hybridized to the specific antibody. Biotin labeling on molecules can be detected by adding a fluorescent molecule or luminescent enzyme that binds to the label, including but not limited to techniques known to those skilled in the art, such as fluorescence, luminescence, FRET, or BRET assays. This readout can be detected using approaches known to those skilled in the art, including but not limited to fluorescence or luminescence detection schemes.

[0158] In some embodiments, the solubilization solution is labeled with biotin to produce a biotinylated compound. In some embodiments, streptavidin is added to bind to the biotinylated compound. In some embodiments, streptavidin is a monomer. In some embodiments, the biotinylated compound is produced by click chemistry. In some embodiments, the compound is labeled by click chemistry after treatment with ARCS, cell isolation, and cell lysis. In some embodiments, the click chemistry reagent contains picolyl azide.

[0159] As used herein, the term “click chemistry” refers to the Huisgen cycloaddition reaction or the 2,3-dipolar cycloaddition reaction between an azide and a terminal alkyne, forming a 1,2,4-triazole. As used herein, the term “cycloaddition reaction” refers to a chemical reaction in which two or more π-electron systems (e.g., unsaturated molecules or unsaturated moieties of the same molecule) are joined to form a cyclic product with a net decrease in bond multiplicity. In a cycloaddition reaction, a new sigma bond is formed using π electrons. The product of the π electrons is called an “adduct” or “cycloadduct.” Different types of cycloaddition reactions are known in the art, including but not limited to [3+2] cycloaddition reactions and Diels-Alder reactions. [3+2] cycloaddition reactions, also called 2,3-dipolar cycloaddition reactions, occur between a 1,3-dipolar and a parent dipolar and are commonly used to construct five-membered heterocycles. The term "[3+2] cycloaddition reaction" also includes "copper-free" [3+2] cycloaddition reactions between azides, cyclooctin derivatives and difluorocyclooctin, as described by Bertozzi et al., J.Am.Chem.Soc., 2004, 126:15046-15047. Any reagent that can be used to facilitate the hysgen cycloaddition reaction can be used as the click chemistry reagent. In some embodiments, the click chemistry reagent comprises pyridyl azide. In some embodiments, the click chemistry reagent comprises pyridyl azide. Any isomer of pyridyl azide may be used, but is not limited to these.

[0160] In some embodiments, ARCS may associate with or bind to one or more radioactive materials or detectable agents. These agents may include a variety of small organic molecules, inorganic compounds, nanoparticles, enzymes or enzyme substrates, fluorescent materials, luminescent materials (e.g., luminol), bioluminescent materials (e.g., luciferase, luciferin, and aequorin), chemiluminescent materials, radioactive materials (e.g., 18 F, 67 Ga, 81m Kr, 82 Rb, 111 In, 123 I, 133 Xe,201 Tl, 125 I, 35 S, 14 C, 3 H or 99m Tc(e.g., pertechnetic acid (technetic acid (VII), TcO4) - Examples of optically detectable labels include 4-acetamido-4'-isothiocyanatostilbene-2,2'disulfonic acid, acridine and its derivatives (e.g., acridine and acridine isothiocyanate), 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS), 4-amino-N-[3-vinylsulfonyl)phenyl]naphthalimide-3,5 Disulfonates, N-(4-anilino-L-naphthyl)maleimide, anthranilamides, BODIPY, Brilliant Yellow, coumarins and derivatives (e.g., coumarin, 7-amino-4-methylcoumarin (AMC, coumarin 120), and 7-amino-4-trifluoromethylcoumarin (coumarin 151)), cyanine dyes, cyanosine, 4',6-diaminidino-2-phenylindole (DAPI), 5'5'-dibromopyrogallol-sulfonaphthalein (bromopyrogallol red) 7-Diethylamino-3-(4'-Isothiocyanatophenyl)-4-methylcoumarin, diethylenetriaminepentaacetate, 4,4'-Diisothiocyanatodihydrostilbene-2,2'-disulfonic acid, 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acid, 5-[dimethylamino]naphthalene-1-sulfonyl chloride (DNS, danyl chloride), 4-dimethylaminophenylazophenyl-4'-isothiocyanate (DABITC), eosin and derivatives (e.g., eosin and eosin isothiocyanate), erythrosine and derivatives (e.g., erythrosine B and erythrosine isothiocyanate), ethidium, fluorescein and derivatives (e.g., 5-carboxyfluorescein Lecein (FAM), 5-(4,6-dichlorotriazine-2-yl)aminofluorescein (DTAF), 2',7'-dimethoxy-4'5'-dichloro-6-carboxyfluorescein, fluorescein isothiocyanate, fluorescein-5-(and-6)-isothiocyanate (QFITC ​​or XRITC) and fluoreskamin), 2-[2-[3-[[1,3-dihydro-1,1-dimethyl-3-(3-sulfopropyl)-2-benzo[e]indole-2-ylidene]ethylidene]-2-[4-(ethoxycarbonyl)-1-piperazinyl]-1-cyclopenten-1-yl]ethenyl]-1,1-dimethyl-3-(3-sulfolpropyl)-1-benzo[e]indrolium hydroxide, intramolecular salt, N,Compounds containing N-diethylethaneamine (1:1) (IR144), 5-chloro-2-[2-[3-[(5-chloro-3-ethyl-2(3H)-benzothiazole-ylidene)ethylidene]-2-(diphenylamino)-1-cyclopenten-1-yl]ethenyl]-3-ethylbenzothiazolium perchlorate (IR140), malachite green isothiocyanate, 4-methylumbelliferone orthocresolphthalein, nitrotyrosine, pararosanilin, phenol red, β-phycoerythrin, oftadialdehyde, pyrene and derivatives (e.g., pyrene, pyrene butyrate and succinimidyl 1-pyrene), butyrate quantum dots, Reactive Red 4 (CIBACRON™ Brilliant Red 3B-A), rhodamine and derivatives (e.g., 6-carboxy-X-rhodamine (ROX), 6-carboxyrhodamine (R6G), lysamine rhodamine B sulfonyl chloride rhodamine (Rhod), rhodamine B, rhodamine 123, rhodamine X isothiocyanate, sulforhodamine B, sulforhodamine 101, sulforhodamine chloride sulfonyl derivative 101 (Texas Red, Examples include N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA), tetramethylrhodamine and tetramethylrhodamine isocyanate (TRITC), riboflavin, rosolic acid, terbium chelate derivatives, cyanine-3 (Cy3), cyanine-5 (Cy5), cyanine-5.5 (Cy5.5), cyanine-7 (Cy7), IRD 700, IRD 800, Alexa 647, La Jolta Blue, phthalocyanine, and naphthalol cyanine.

[0161] In some embodiments, the detectable agent may be an undetectable precursor that becomes detectable upon activation (e.g., a fluorescent tetrazine-fluorophore construct (e.g., tetrazine-BODIPY FL, tetrazine-Oregon Green 488, or tetrazine-BODIPY TMR-X)) or an enzyme-activating fluorescence generator (e.g., PROSENSE® (Vis Medical))).

[0162] IV. Definition As used herein, the term "ARCS" refers to any therapeutic conjugate formed by linking FCB and CLM by a linker. In some embodiments, ARCS can form covalent bonds with one or more targets, such as nucleotides, oligonucleotides, peptides, or proteins. In some embodiments, the covalent bonds are formed in aqueous solution at a therapeutic dose of 10 mM within 48 hours at a temperature of 0–50°C.

[0163] As used herein, the term “FCB” refers to a therapeutic modality which may be a known drug, diagnostic compound, drug candidate, functional fragment, and / or any combination thereof. An FCB includes isotopes comprising free acid and free base forms, optical isomers and tautomers, radioisotopes and pharmaceutically acceptable salts of a drug, its prodrug or fragment. An FCB may be a small molecule, protein, peptide, lipid, carbohydrate, sugar, nucleic acid, or a combination thereof. In some embodiments, an FCB is a nucleic acid, including but not limited to DNA or RNA. An FCB may be a therapeutic agent, such as an anticancer agent, an antineurodegenerative agent, an autoimmune agent, and an anti-aging agent, but is not limited to these. An FCB may be non-covalently bound to a biological target. In some embodiments, an FCB may be a functional fragment of a drug. As used herein, the term “functional fragment” refers to a portion of a drug or its derivatives or analogs that can induce the desired effect of the drug. In some embodiments, an FCB may contain an alkyne functional group. In some embodiments, an FCB may not contain an alkyne functional group.

[0164] As used herein, the term "CLM" refers to any covalent modality capable of forming a covalent bond with a biological target. The CLM may be linked to the FCB by a bond or a linker. The CLM may contain one or more chemical moieties capable of forming a covalent bond with a biological target. The chemical moieties may be electrophilic or nucleophilic groups.

[0165] As used herein, the term "linker" refers to an organic portion that connects two parts of a compound. The linker may be an external linker or an internal linker. An external linker can connect the FCB portion and the CLM portion. An internal linker can be used to bond the CLM portion. In certain embodiments, the CLM may include an internal linker or a spacer. The internal linker or spacer may combine or bond the two parts of the CLM. External or internal linkers may be bonded, substituted, and unsubstituted C1-C 30 Alkyl, substituted, and unsubstituted C2-C 30 Alkenyl, substituted and unsubstituted C2-C 30 Alkynyl, substituted and unsubstituted C3-C 30 Cycloalkyl, substituted, and unsubstituted C 1~ C 30 Heterocycloalkyl, substituted and unsubstituted C3-C 30 Cycloalkenyls, substituted and unsubstituted C1-C 30 The linker can be selected from the group consisting of heterocycloalkenyls, substituted and unsubstituted aryls, and substituted and unsubstituted heteroaryls. The linker may be cleavable or uncleavable.

[0166] As used herein, the term “biological target” refers to any target to which FCBs bind non-covalently to produce a therapeutic effect. CLMs bind covalently to biological targets. In some embodiments, the biological target is a protein.

[0167] As used herein, the term “toxicity” refers to the ability of a substance or composition to be harmful or toxic to cells, living tissues, or the cellular environment. “Low toxicity” refers to the attenuated ability of a substance or composition to be harmful or toxic to cells, living tissues, or the cellular environment. Such attenuation or low toxicity may be relative to a standard measure, to treatment, or to the absence of treatment.

[0168] As used herein, the term “compound” means including all stereoisomers, geometric isomers, tautomers, and isotopes of the illustrated structure. In some embodiments, the compound is used interchangeably with ARCS. Therefore, as used herein, ARCS also means including all stereoisomers, geometric isomers, tautomers, and isotopes of the illustrated structure. As used herein, FCB and CLM also mean including all stereoisomers, geometric isomers, tautomers, and isotopes of the illustrated structure.

[0169] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of this disclosure containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically active starting materials, such as resolution of racemic mixtures or stereoselective synthesis, are known in the art. Many geometric isomers, such as olefins and C=N double bonds, can also be present in the compounds described herein, and all such stable isomers are intended in this disclosure. Cis and trans geometric isomers of the compounds of this disclosure are described and may be isolated as mixtures of isomers or as separated isomers.

[0170] The compounds of this disclosure also include tautomers. Tautomers arise from the swap of single bonds with adjacent double bonds and the simultaneous transfer of protons. Tautomers include prototropic tautomers, which are isomeric protonation states having the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and protons such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole 1H, and 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomers may be in equilibrium or sterically locked into one form by appropriate substitution.

[0171] The compounds of this disclosure also include all isotopes of the atoms that occur in the intermediate or final compounds. “Isotopes” refer to atoms that have the same atomic number but different mass numbers resulting from different numbers of neutrons in their nuclei. For example, isotopes of hydrogen include tritium and deuterium.

[0172] The compounds and salts of this disclosure can be prepared in combination with a solvent or water molecules to form solvates and hydrates by routine methods.

[0173] As used herein, the terms “subject” or “patient” refer to any organism to which particles may be administered, for example, for experimental, therapeutic, diagnostic, and / or prophylactic purposes. Typical subjects include animals (mammals such as mice, rats, rabbits, guinea pigs, cattle, pigs, sheep, horses, dogs, cats, hamsters, llamas, non-human primates, and humans).

[0174] As used herein, the terms “treat” or “prevent” can include preventing the onset of a disease, disorder, or condition in an animal that is suspected of being susceptible to a disease, disorder, or condition but has not been diagnosed with the disease, disorder, or condition; inhibiting a disease, disorder, or condition, for example, inhibiting its progression; or mitigating a disease, disorder, or condition, for example, causing regression of the disease, disorder, or condition. Treating a disease, disorder, or condition can include improving at least one symptom of a particular disease, disorder, or condition, even if the underlying pathophysiology is not affected, for example, treating pain in a subject by administering an analgesic, even if the analgesic does not treat the cause of the pain.

[0175] As used herein, “target” means the site to which ARCS, FCB and / or CLM bind. The target may be either in vivo or in vitro. In certain embodiments, the target may be cancer cells found in leukemia or tumors (e.g., tumors of the brain, lungs (small cell and non-small cell), ovaries, prostate, breast and colon, as well as other cancers and sarcomas). The target may be a type of tissue, such as nerve tissue, intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovaries, lungs, bone marrow, or breast tissue.

[0176] "Target cells" that can function as targets for therapeutic conjugates are generally animal cells, such as mammalian cells. The methods of the present invention can be used to modify the cellular function of living cells that form part of animal tissue or are otherwise present in animal tissue in vitro, i.e., in cell culture or in vivo. Accordingly, target cells can include, for example, blood, lymphoid tissue, cells lining the digestive tract such as oral and pharyngeal mucosa, cells that form the ciliary body of the small intestine, cells lining the large intestine, cells lining the animal respiratory system (nasal cavity / lungs) (which may be in contact by inhalation of the target), skin / epidermal cells, vaginal and rectal cells, placental cells, and visceral cells including the so-called blood-brain barrier.

[0177] The term "therapeutic effect" is as recognized in the art and refers to a topical or systemic effect caused by a pharmacologically active substance in animals, particularly mammals, and more specifically, humans. Therefore, the term means any substance intended for use in the diagnosis, cure, alleviation, treatment or prevention of disease, or in enhancing desirable physical or mental development and condition in animals or humans.

[0178] The term "modulation" is recognized in the art and refers to upmodulation (i.e., activation or stimulation), downmodulation (i.e., inhibition or suppression) of a response, or a combination of the two or separately.

[0179] As used herein, “enteral administration” means administration by any method other than the gastrointestinal tract (enterum) or a non-invasive local route. For example, parenteral administration may include administration to a patient by intravenous, intradermal, intraperitoneal, intrapleural, intratracheal, intraosseous, intracerebral, intrathecal, intramuscular, subcutaneous, subconjunctival, injection, and infusion.

[0180] As used herein, “topical administration” means non-invasive administration to the skin, orifice or mucous membrane. Topical administration can be administered locally, i.e., without systemic exposure. It can provide a local effect to the area of ​​application. Topical formulations can provide systemic effects through adsorption to the bloodstream of the individual. Topical administration methods include, but are not limited to, skin and transdermal administration, buccal administration, intranasal administration, vaginal administration, intravesical administration, ocular administration, and rectal administration.

[0181] As used herein, "enteral administration" means administration via absorption through the gastrointestinal tract. Enteral administration may include oral and sublingual administration, gastric administration, or rectal administration.

[0182] As used herein, “pulmonary administration” means administration to the lungs by inhalation or intratracheal administration. As used herein, the term “inhalation” means the inhalation of air into the alveoli. Air intake may occur through the mouth or nose.

[0183] Where used interchangeably herein, the terms “sufficient” and “effective” refer to the amount (e.g., mass, volume, dose, concentration, and / or duration) required to achieve one or more desired results. “Therapeutic effective dose” is the minimum concentration required to produce a measurable improvement or prevention of at least one symptom-specific condition or disorder, a measurable increase in life expectancy, or generally an improvement in the patient’s quality of life. Thus, the therapeutic effective dose depends on the specific bioactive molecule and the specific condition or disorder being treated. The therapeutic effective doses of many activators, such as antibodies, are known in the art. For example, to treat a particular disorder, the therapeutic effective dose of the compounds and compositions described herein can be determined by art that is within the scope of the art of those skilled in the art, such as physicians.

[0184] The term "prodrug" refers to a drug containing nucleic acids or proteins that are converted into a biologically active form in vitro and / or in vivo. In some situations, prodrugs may be useful because they are easier to administer than the parent compound. For example, a prodrug may be biologically available by oral administration, while the parent compound is not. Prodrugs may also have improved solubility in pharmaceutical compositions compared to the parent drug. Prodrugs may be converted to the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. (Harper, NJ (1962) Drug) Latentiation in Jucker、d.Progress in Drug Research、4:221-294;Molozowichら(1977)Application of Physical Organic Principles to Prodrug Design in E.B.Roche ed.Design of Biopharmaceutical Properties through Prodrugs and Analogs, APhA;Acad.Pharm.Sci.;E.B.Roche、ed.(1977)Bioreversible Carriers in Drug in Drug Design、Theory and Application、APhA;H.Bundgaard、ed.(1985)Design of Prodrugs、Elsevier、Wangら(1999)Prodrug approachs to the professional of peptide drug、Curr.Pharm.Design.5(4):265-287;Paulettiら(1997)Improvement in peptide bioavailability: Peptidomimetics and Prodrug Strategies、Adv.Drug.Delivery Rev.27:235-256;Mizenら(1998)。The Use of Esters as Prodrugs for Oral Delivery of β-Lactam antibiotics、Pharm.Biotech.11:345-365;Gaignaultら(1996)Designing Prodrugs and Bioprecursors I.Carrier Prodrugs、Pract.M ed.Chem.671-696;M.Asgharnejad(2000)。Improving Oral Drug Transport Via Prodrugs、in G.L.Amidon,P.I.Lee and E.M.Topp、Eds.、Transport Processes in Pharmaceutical Systems、Marcell Dekker、p.185-218;Balantら(1990)Prodrugs for the improvement of drug absorption via different routes of administration、Eur.J.Drug Metab.Pharmacokinet.、15(2):143-53;Balimane and Sinko(1999)。Involvement of multiple transporters in the oral absorption of nucleoside analogs、Adv.Drug Delivery Rev.、39(1-3):183-209;Browne(1997)。Fosphenytoin(Cerebyx)、Clin.Neuropharmacol.20(1):1-12;Bundgaard(1979)。Bioreversible derivatization of drugs--principle and applicability to improve the therapeutic effects of drugs、Arch.Pharm.Chemi.86(1):1-39;H.Bundgaard、ed.(1985)Design of Prodrugs、Elsevier、Fleisherら(1996)Improved oral drug delivery:solubility limitations overcome by the use of prodrugs、Adv.Drug Delivery Rev.19(2):115-130;Fleisherら(1985)Design of prodrugs for imprograms for imprograms gastrounic absorption by intestinal enzyme targeting、Methods Enzymol.112:360-81;Farquhar Dら(1983)Biologically Reversible Phosphate-Protective Groups、J.Pharm.Sci.,72(3):324-325;Han,H.K.ら(2000)Targeted prodrug design to optimize drug delivery、AAPS PharmSci.,2(1):E6;Sadzuka Y.(2000)Effective prodrug liposome and conversion to active metabolite、Curr.Drug Metab.、1(1):31-48;.M.Lambert(2000)Rationale and applications of lipids as prodrug carriers、Eur.J.Pharm.Sci.、11 Suppl.2:S15-27;Wangら(1999)Prodrug approaches to the improved delivery of peptide drugs.Curr.Pharm.Des.,5(4):265-87。

[0185] The term "pharmaceutically acceptable," as used herein, refers to compounds, materials, compositions, and / or dosage forms that are within the bounds of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in accordance with the guidelines of agencies such as the U.S. Food and Drug Administration, and that are commensurate with a reasonable benefit / risk ratio. As used herein, "pharmaceutically acceptable carrier" refers to all components of a pharmaceutical formulation that facilitate the delivery of a composition in vivo. Pharmaceutically acceptable carriers include, but are not limited to, diluents, preservatives, binders, lubricants, disintegrants, swelling agents, fillers, stabilizers, and combinations thereof.

[0186] As used herein, the term “molecular weight” generally refers to the mass or average of a material. This refers to mass. In the case of polymers or oligomers, molecular weight can refer to the relative average chain length or relative chain mass of the bulk polymer. In practice, the molecular weight of polymers and oligomers can be estimated or characterized by a variety of methods, including gel permeation chromatography (GPC) or capillary viscosity. GPC molecular weight is reported as weight-average molecular weight (Mw), as opposed to number-average molecular weight (Mn). Capillary viscosity provides an estimate of molecular weight as intrinsic viscosity determined from a diluted polymer solution using a specific set of concentration, temperature, and solvent conditions.

[0187] As used herein, the term “small molecule” generally refers to an organic molecule with a molecular weight of less than 2000 g / mol, less than 1500 g / mol, less than 1000 g / mol, less than 800 g / mol, or less than 500 g / mol. Small molecules are nonpolymers and / or non-oligomers.

[0188] The term "alkyl" refers to a saturated aliphatic radical, including linear alkyl groups, branched alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups.

[0189] In some embodiments, linear or branched alkyl groups have 30 or fewer carbon atoms in their skeleton (for example, a linear group may have C 1- C 30 The branching chain is C 3- C 30 ), having 20 or fewer carbon atoms, 12 or fewer carbon atoms, or 7 or fewer carbon atoms. Similarly, in some embodiments, cycloalkyls have 3 to 10 carbon atoms in the ring structure, for example, having 5, 6, or 7 carbon atoms in the ring structure. As used herein, in the examples and throughout the claims, the term “alkyl” (or “lower alkyl”) is intended to include both “unsubstituted alkyl” and “substituted alkyl,” the latter referring to an alkyl moiety having one or more substituents that substitute hydrogen at one or more carbon atoms in the hydrocarbon skeleton. Such substituents include, but are not limited to, halogens, hydroxyls, carbonyls (such as carboxyls, alkoxycarbonyls, formyl or acyls), thiocarbonyls (such as thioesters, thioacetates or thioformates), alkoxyls, phosphoryls, phosphoric acids, phosphonic acids, phosphinic acids, aminos, amides, amidines, imines, cyanos, nitros, azides, sulfhydryls, alkylthios, sulfuric acids, sulfonic acids, sulfamoyls, sulfonamidesulfonyls, heterocyclyls, aralkyls, or aromatic or heteroaromatic moieties.

[0190] Unless otherwise specified, as used herein, “lower alkyl” means an alkyl group as defined above, but having 1 to 10 carbon atoms or 1 to 6 carbon atoms in its skeletal structure. Similarly, “lower alkenyl” and “lower alkynyl” have similar chain lengths. Throughout this application, preferred alkyl groups are lower alkyl groups. In some embodiments, substituents designated as alkyl in this specification are lower alkyl groups.

[0191] Those skilled in the art will understand that a substituted portion on a hydrocarbon chain may, where appropriate, be substituted itself. For example, substituents on substituted alkyls may include halogens, hydroxyl, nitro, thiol, amino, azide, imino, amide, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl and sulfonates), and silyl groups, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, etc. Cycloalkyls may be substituted in a similar manner.

[0192] As used herein, the term “heteroalkyl” refers to a linear, branched, or cyclic carbon-containing radical or combination thereof containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, and S. Examples include e, B, and S, where the phosphorus and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. Heteroalkyls can be substituted for alkyl groups as defined above.

[0193] The term "alkylthio" refers to an alkyl group having a sulfur radical bonded to it, as defined above. In some embodiments, the "alkylthio" portion is represented by one of -S-alkyl, -S-alkenyl, and -S-alkynyl groups. Representative alkylthio groups include methylthio and ethylthio. The term "alkylthio" also encompasses cycloalkyl groups, alkene groups, cycloalkene groups, and alkyne groups. "Arylthio" refers to an aryl group or heteroaryl group. Alkylthio groups can be substituted for alkyl groups as defined above.

[0194] The terms "alkenyl" and "alkynyl" refer to unsaturated aliphatic groups that are similar to the alkyl groups mentioned above in terms of length and possible substitutions, but each contains at least one double or triple bond.

[0195] As used herein, the terms “alkoxyl” or “alkoxy” refer to an alkyl group as defined above, having an oxygen radical bonded to it. Typical alkoxyl groups include methoxy, ethoxy, propyloxy, and tert-butoxy. “Ether” is a hydrocarbon covalently bonded by oxygen. Therefore, substituents on an alkyl group that make an alkyl group an ether are alkoxyl or similar to alkoxyl, and can be represented, for example, by one of -O-alkyl, -O-alkenyl, and -O-alkynyl. Aroxy may also be represented by -O-aryl or O-heteroaryl, as defined below. Alkoxy and aroxy groups can be substituted for alkyl groups as described above.

[0196] The terms "amine" and "amino" are recognized in the art and refer to both unsubstituted and substituted amines, for example, in the general formula: [ka] This is the part that can be represented by R9, R 10 , and R' 10 These are, independently, hydrogen, alkyl, alkenyl, and -(CH2) m -R8 or R9 and R 10 These, together with the N atoms to which they are bonded, complete a heterocycle having 4 to 8 atoms in the ring structure. R8 represents an aryl, cycloalkyl, cycloalkenyl, heterocycle, or polycycle, and m is zero or an integer in the range of 1 to 8. In some embodiments, R9 or R 10 Only one of them can be a carbonyl group, for example, R9, R 10 Oh Nitrogen does not combine with the amine to form an imide. In yet another embodiment, the term "amine" does not include amides, for example, R9 and R 10One of them represents a carbonyl group. In additional embodiments, R9 and R 10 (and optionally R' 10 ) each independently represents hydrogen, alkyl or cycloalkyl, alkenyl or cycloalkenyl or alkynyl. Therefore, as used herein, the term "alkylamine" is an amine group having a substituted (as described above for alkyl) or unsubstituted alkyl bonded to it, i.e., R9 and R 10 This means that at least one of them is an alkyl group.

[0197] The term "amide" is recognized in the art as an amino-substituted carbonyl, and its general formula is: [ka] It includes parts that can be represented by R9 and R 10 It is defined as described above.

[0198] As used herein, "aryl" means C5-C 10 This refers to member-based aromatic, heterocyclic, fusion aromatic, fusion heterocyclic, diaromatic, or diheterocyclic systems. In a broader sense, as used herein, "aryl" includes 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, and 10-membered monocyclic aromatic groups that may contain 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Aryl groups having heteroatoms in their ring structure are also referred to as "aryl heterocyclic" or "heteroaromatic." Aromatic rings may also include, but are not limited to, halogens, azides, alkyls, aralkyls, alkenyls, alkynyls, cycloalkyls, hydroxyls, alkoxyls, aminos (or quaternized aminos), nitros, sulfhydryls, iminos, amides, phosphonic acids, phosphinates, carbonyls, carboxyls, silyls, ethers, alkylthios, sulfonyls, sulfonamides, ketones, aldehydes, esters, heterocycles, aromatic rings or heteroaromatic moieties, -CF3, -CN; and combinations thereof.

[0199] The term "aryl" also refers to a macrocyclic system having two or more cyclic rings, where two or more carbons are common to two adjacent rings (i.e., fusion rings), and at least one of the rings is aromatic, for example, the other cyclic ring or ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclic.Examples of heterocyclic compounds, though not limited to them, include: benzimidazolyl, benzofuranil, benzothiofuranil, benzothiophenyl, benzoxazolyl, benzoxazolinil, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinil, carbazolyl, 4aH-carbazolyl, carborinil, chromanil, chromenil, sinnolinil, decahydroquinolinil, 2H,6H-1,5,2-dithiadinyl, dihydroflo[2,3b]tetrahydrofuranil, furanil, furazanyl Imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolidinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranil, isochromanil, isoindazolyl, isoindonyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthilidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1 ,3,4-Oxadiazolyl, Oxazolidinil, Oxazolyl, Oxadyl, Pyrimidinil, Phenanthronil, Phenadhiazinil, Phenoxatinil, Phenoxadinil, Phthalazinil, Piperadinil, Piperidinil, Piperidonil, 4-Piperidonil, Piperonil, Pteridinil, Prinnil, Pyranil, Pyrazinil, Pyrazolidinil, Pyrazolyl, Pyridadinil, Pyridoxazole, Pyridoimidazole, Pyridhiazole, Pyridinil, Pyridyl, Pyrimidinil, Pyrrolidinil Examples include pyrrolinil, 2H-pyrrolyl, pyrrolyl, quinazolinil, quinolinil, 4H-quinolidinil, quinoxalinil, quinusidinil, tetrahydrofuranil, tetrahydroisoquinolinil, tetrahydroquinolinil, tetrazolyl, 6H-1,2,5-thiadiadinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.One or more rings can be replaced with "aryls" as defined above.

[0200] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group (e.g., an aromatic group or a heteroaromatic group).

[0201] As used herein, the term “carbocyclic” refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

[0202] "Heterocyclic" or "heterocyclic formula," as used herein, refers to a cyclic radical bonded via ring carbon or nitrogen in a monocyclic or bicyclic ring containing 3 to 10 ring atoms, preferably 5 to 6 ring atoms, and composed of 1 to 4 heteroatoms selected from the group consisting of non-peroxide oxygen, sulfur, and N(Y), with Y absent, or H, O, (C1-C 10 ) Alkyl, phenyl, or benzyl, optionally containing 1 to 3 double bonds, and optionally substituted with one or more substituents. Examples of heterocyclic rings While not limited to these, the following are examples of benzimidazolyl, benzofuranil, benzothiofuranil, benzothiophenyl, benzoxazolyl, benzoxazolinil, benzthiazolyl, benztriazolyl, benztetrazolyl, benzsoxazolyl, benzsothiazolyl, benzimidazolinil, carbazolyl, 4aH-carbazolyl, carborinil, chromanil, clomenil, sinnolinil, decahydroquinolinil, 2H,6H-1,5,2-dithiadinyl, dihydroflo[2,3b]tetrahydrofuranil, furanil, furazanil, imi Dazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolidinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranil, isochromanil, isoindazolyl, isoindonyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthilidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3, 4-Oxadiazolyl, oxazolidinil, oxazolyl, oxidyl, pyrimidinil, phenanthronil, phenadhiazinil, phenoxathinil, phenoxadinil, phthalazinil, piperazinil, piperidinil, piperidonil, 4-piperidonil, piperonil, pteridinil, prinil, pyranil, pyrazinil, pyrazolidinil, pyrazolyl, pyrazolyl, pyridazinil, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinil, pyridyl, pyrimidinil, pyrrolidinil, pi Examples include lorinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolidinyl, quinoxalinyl, quinusidinyl, tetrahydrofuranil, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiadinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.The aromatic ring group, as defined above, may be optionally substituted with one or more substituents at one or more positions in, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonic acid, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocycle, aromatic ring or heteroaromatic moiety, -CF3 and -CN.

[0203] The term "carbonyl" is recognized in the art and has the general formula: [ka] It includes a part that can be represented by, where X is a bond, or represents oxygen or sulfur, and R 11 R' represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or alkynyl. 11 X represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or anyle. X is oxygen, and R 11 or R' 11 If X is not hydrogen, the formula represents an "ester". If X is oxygen, R 11 If as defined above, the part is referred to herein as a carboxyl group, and in particular R 11 When X is hydrogen, the formula represents a "carboxylic acid". 11 When X is hydrogen, the formula represents "formic acid". Generally, when the oxygen atom in the above formula is substituted with sulfur, the formula represents a "thiocarbonyl" group. When X is sulfur, R 11 or R' 11 If X is not hydrogen, the formula represents a "thioester". If X is sulfur, R 11 When hydrogen is present, the formula represents "thiocarboxylic acid". X is sulfur, and R' 11 If X is hydrogen, the formula represents "thioformic acid". On the other hand, if X is a bond, R 11 If X is not hydrogen, the above formula represents a "ketone" group. X is a bond, and R 11When the group is hydrogen, the above formula represents an "aldehyde" group.

[0204] As used herein, the term “monoester” refers to an analogue of a dicarboxylic acid, in which one carboxylic acid is functionalized as an ester and the other carboxylic acid is a free carboxylic acid or a salt of a carboxylic acid. Examples of monoesters include, but are not limited to, monoesters of succinic acid, glutamic acid, adipic acid, sulfite, sebacic acid, azeleic acid, oxalic acid, and maleic acid.

[0205] As used herein, the term “heteroatom” means an atom of any element other than carbon or hydrogen. Examples of heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur, and selenium. Other heteroatoms include silicon and arsenic.

[0206] As used herein, the term "nitro" means -NO2; the term "halogen" means -F, -Cl, -Br, or -I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyl" means -SO2-.

[0207] As used herein, the term “substituted” refers to all permissible substituents of the compounds described herein. In its broadest sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents, but not limited to, include halogens, hydroxyl groups, or any other organic group containing any number of carbon atoms, preferably 1 to 14 carbon atoms, and optionally, one or more heteroatoms such as oxygen, sulfur, or nitrogen in linear, branched, or cyclic structural formats. Typical substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, alloxy, substituted alloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amide, substituted amide, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C 3- C 20 Circular, substitution C 3- C 20 This includes cyclic, heterocyclic, substituted heterocyclic, amino acid, peptide, and polypeptide groups.

[0208] Heteroatoms such as nitrogen may have hydrogen substituents and / or any acceptable substituents of the organic compounds described herein that satisfy the valence of the heteroatom. Naturally, “substituted” or “substituted” implicitly includes the condition that such substitutions comply with the acceptable valences of the substituted atom and substituent, and that the substitutions result in stable compounds, i.e., compounds that do not undergo spontaneous transformation by reconstitution, cyclization, or removal, etc.

[0209] In broad embodiments, acceptable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, for example, those described herein. Acceptable substituents may be one or more identical or different substituents for a given organic compound. Heteroatoms such as nitrogen may have hydrogen substituents and / or any acceptable substituents of the organic compound described herein that satisfy the valence of the heteroatom.

[0210] In various embodiments, the substituents are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamic acid, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, each optionally substituted with one or more suitable substituents. In some embodiments, substitutions are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamic acid, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphoric acid, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, and alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamic acid, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphoric acid, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone may be further substituted with one or more suitable substituents.

[0211] Examples of substituents, though not limited to them, include halogens, azides, alkyls, aralkyls, alkenyls, alkynyls, cycloalkyls, hydroxyls, alkoxyls, aminos, nitrosulfhydryls, iminos, amides, phosphonic acids, phosphinates, carbonyls, carboxyls, silyls, ethers, alkylthios, sulfonyls, sulfonamides, ketones, aldehydes, thioketones, esters, heterocyclyls, -CNs, aryls, aryloxys, perhalalkoxys, aralkoxys, heteroaryls, heteroaryloxys, heteroarylalkyls, and hetero Examples include aralkoxy, azide, alkylthio, oxo, acylalkyl, carboxyester, carboxamide, acyloxy, aminoalkyl, alkylaminoaryl, alkylaryl, alkylaminoalkyl, alkoxyaryl, arylamino, aralkylamino, alkylsulfonyl, carboxamidealkylaryl, carboxamidearyl, hydroxyalkyl, haloalkyl, alkylaminoalkylcarboxy, aminocarboxamidealkyl, cyano, alkoxyalkyl, perhaloalkyl, and arylalkyloxyalkyl. In some embodiments, the substituent is selected from cyano, halogen, hydroxyl, and nitro.

[0212] The terms “polypeptide,” “peptide,” and “protein” generally refer to polymers of amino acid residues. As used herein, these terms also apply to amino acid polymers in which one or more amino acids are chemical analogs or modified derivatives of corresponding naturally occurring amino acids. The term “protein,” as commonly used herein, refers to a polymer of amino acids linked together by peptide bonds, forming polypeptides whose chain length is sufficient to create tertiary and / or quaternary structures. The term “protein” excludes small peptides by definition, as small peptides lack the higher-order structures necessary to be considered proteins.

[0213] A “functional fragment” of a protein, polypeptide, or nucleic acid is a protein, polypeptide, or nucleic acid whose sequence is not identical to that of the full-length protein, polypeptide, or nucleic acid, but which retains at least one function of the full-length protein, polypeptide, or nucleic acid. A functional fragment may have more, fewer, or the same number of residues as the corresponding native molecule, and / or may contain one or more amino acid or nucleotide substitutions. Methods for determining the function of a nucleic acid (e.g., coding function, ability to hybridize to another nucleic acid) are well known in the art. Similarly, methods for determining the function of a protein are well known. For example, the DNA-binding function of a polypeptide can be determined, for example, by filter binding, electrophoretic mobility shift, etc. This can be determined by immunoprecipitation assays. DNA cleavage can be assayed by gel electrophoresis. The ability of a protein to interact with another protein can be determined, for example, by co-immunoprecipitation, bihybrid assays, or complementarity, for example, genetically or biochemically. See, for example, Fields et al. (1989) Nature 340:245-246, U.S. Patent No. 5,585,245 and PCT Patent No. 98 / 44350.

[0214] The term "pharmaceutically acceptable counterion" refers to a pharmaceutically acceptable anion or cation. In various embodiments, a pharmaceutically acceptable counterion is a pharmaceutically acceptable ion. For example, pharmaceutically acceptable counterions include citrate, malate, acetate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphoric acid, phosphate, isonicotinic acid, acetate, lactate, salicylate, tartrate, oleate, tannate, pantothenate, bicarbonate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, and benzenesulfonate. The counterions are selected from folates, p-toluenesulfonates, and pamoates (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). In some embodiments, pharmaceutically acceptable counterions are selected from chlorides, bromides, iodides, nitrates, sulfates, disulfates, phosphates, acid phosphates, citrates, malates, acetates, oxalates, acetates, and lactates. In certain embodiments, pharmaceutically acceptable counterions are selected from chlorides, bromides, iodides, nitrates, sulfates, disulfates, and phosphates.

[0215] The term "pharmaceutically acceptable salt" refers to a salt of an acidic or basic group that may be present in the compounds used in the compositions of the present invention. Compounds included in the compositions of the present invention that are basic in nature can form a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are acids that form non-toxic acid addition salts, i.e., salts containing a pharmaceutically acceptable anion, and are not limited to, sulfates, citrates, malates, acetates, oxalates, chlorides, bromides, iodides, nitrates, sulfates, disulfates, phosphates, acid phosphates, isonicotinates, acetates, lactates, salicylates, citrates, tartrates, oleates, tannates, pantothenates, hydrogen tartrates, ascorbic acid, succinates, maleates, gentisates, fumarates, glucons, glucarones, saccharates, formates, benzoates, and glucons. The compounds in the compositions of the present invention, which include tamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate) salts), can form pharmaceutically acceptable salts having various amino acids in addition to the acids described above. Compounds in the compositions of the present invention, which are acidic in nature, can form basic salts with various pharmaceutically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, particularly calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.

[0216] When the compounds described herein are obtained as acid addition salts, the free base can be obtained by basicizing a solution of the acid acid. Conversely, when the product is a free base, the addition salt, in particular a pharmaceutically acceptable addition salt, can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid, following conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art are aware of the various synthetic methodologies that can be used to prepare non-toxic, pharmaceutically acceptable addition salts.

[0217] The pharmaceutically acceptable salts are 1-hydroxy-2-naphthic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutamic acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, bromide It can be produced in chloroacetic acid, hydrochloric acid, isethionic acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucus, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phosphoric acid, proprionic acid, pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid, thiocyanic acid, toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid.

[0218] As used herein, the term “assay” refers to a set of activities related to the reported results, which may include, but are not limited to, cell seeding, preparation of test material, infection, lysis, analysis, and calculation of results.

[0219] As used herein, the term “detectable response” refers to the occurrence or change of a signal that can be detected directly or indirectly by either observation or measurement means. Typically, a detectable response is the occurrence of a signal, where a fluorophore is inherently fluorescent and does not produce a change in signal when bound to a metal ion or biological compound. Alternatively, a detectable response is an optical response resulting in a change in wavelength distribution pattern or absorbance or fluorescence intensity, or a change in light scattering, fluorescence lifetime, fluorescence polarization, or a combination of the above parameters. Other detectable responses include, for example, chemiluminescence, phosphorescence, radiation from radioisotopes, magnetic attraction, and electron density.

[0220] Naturally, the following embodiments are illustrative but not limiting to the present disclosure. Various other embodiments and modifications to the foregoing description and examples will be apparent to those skilled in the art after reading the present disclosure without departing from the spirit and scope of the present disclosure, and all such examples or modifications are intended to be included in the appended claims. All publications and patents referenced herein are incorporated herein in their entirety by reference. [Examples]

[0221] Example 1: General synthesis of ARCS The ARCSs of this disclosure can be synthesized by those skilled in the art using common chemical synthesis principles and techniques. In a reasonable approach, ARCSs are constructed from their individual components, namely therapeutic modalities, optional linkers, and covalent modalities. The components may be covalently bonded to one another via functional groups, as known in the art, and such functional groups may be present on the components or introduced onto them using one or more steps. Functional groups that can be used to covalently bond components together to produce ARCSs include, but are not limited to, hydroxyl, sulfhydryl, or amino groups. Certain parts of different components that are modified to provide covalent bonding are selected, for example, for a covalent modality, so as not to substantially adversely affect the desired bonding activity of that component, and the areas that do not affect covalent activity are modified so as to retain a sufficient amount of the desired activity. Where necessary and / or desired, certain parts on a component may be protected using blocking groups known in the art; see, for example, Green & Wuts, Protective Groups in Organic Synthesis (John Wiley & Sons) (1991).

[0222] Alternatively, ARCS may be prepared using well-known combinatorial methods to create a large library of ARCS, which may then be screened to identify molecules that form covalent bonds with targets having a desired pharmacokinetic profile.

[0223] Example 2: General synthesis of compounds 1-1 to 1-172 Compounds 1-1 to 1-172 of this disclosure can be synthesized by those skilled in the art using common chemical synthesis principles and techniques. The chemistry is as described in Example 1 of U.S. Patent No. 9,724,352B2, which is incorporated herein by reference in its entirety.

[0224] Compound 1-IV, a precursor to many compounds 1-1 to 1-172, can be prepared as shown in Scheme 1. As described in U.S. Patent No. 9,724,352, the starting pyrrole (1-I) can be reacted with the aldehyde (1-II) to form the intermediate (1-III). Morphorino compound 1-IV is formed by reducing compounds (1-III) with phosphorus oxychloride to provide a Cl leaving group, which can then be substituted by the addition of morpholine. If R6' in 1-IV is a nitro group, it can be reduced to the corresponding NH2 group by reacting with C / Pd under an H2 atmosphere. The amine can then be further reacted with a suitable ester (e.g., dimethyl carbonate) or activated ester (e.g., methyl chloroformate) to form the methyl carbamate of the compound, or with an isocyanate (e.g., methyl isocyanate) to obtain urea compound 1-V. Scheme 1 [ka]

[0225] To obtain piperazinyl compounds (e.g., compounds 1-11 and 1-12), it may be advantageous to modify the methyl ester moiety of compound 1-IV and then convert R6' to the final R6 group. As mentioned above, if R6' of 1-IV is a nitro group, it can be reduced to the corresponding NH2 group by Pd / C under an H2 atmosphere. As shown in Scheme 2, the methyl ester of 1-IV is hydrolyzed (e.g., 2M aqueous NaOH in EtOH), and the resulting acid is t in the presence of a base (e.g., K2CO3) By reacting with ert-butylpiperazine-1-carboxylic acid (1-BOC-piperazine), a BOC-piperazinyl compound can be obtained. Then, the carbonyl moiety remaining after the addition of piperazinyl (e.g., borane-dimethyl sulfide) is reduced to obtain a protected piperazinyl compound 1-VI. Next, the amine group of compound 1-VI is further reacted with a suitable ester (e.g., dimethyl carbonate) or an activated ester (e.g., methyl chloroformate) to form the methyl carbamate of compound 1-V, or reacted with an isocyanate (e.g., methyl isocyanate) to obtain urea compound 1-VII. A (See, for example, the groups of compounds 1-105 and 1-106.) The amine group of compound 1-VI can also be protected if it is the final desired group (not shown) (see compound 1-121). After the desired X group is added, the amino protecting group can be removed to obtain the final product. For example, by using N-acetyl-piperazine instead of N-BOC-piperazine, an N-acetyl protecting group can be obtained in compound 1-VI. The amino group of compound 1-VI can then be protected with an N-BOC group, which is more stable than N-acetyl. Compound 1-VI can then be modified with the X group and the N-BOC can be removed to leave the final amino group. Scheme 2 [ka]

[0226] Additional starting compounds 1-III A-C (See compounds 1-101 to 1-172) can be prepared as shown in Scheme 3. These starting compounds can then be modified as described herein. Scheme 3 [ka]

[0227] The X group of this disclosure (see, for example, compounds 1-101 to 1-172) can be bonded as shown in Scheme 4. The BOC protecting group of compound 1-VII (or the acetyl described above) can be removed via acid hydrolysis (e.g., HCl in methanol) to obtain compound 1-VIII. The piperazinyl group of compound 1-VIII can then be bonded appropriately in the presence of a base (e.g., K2CO3 or dimethylaminopyridine). Esters (e.g., 1-IX and 1-IX) A For this, CH2=CHC(O)(CH2)2C(O)OCH3 or its acid or activating acid can be used) to react with the final compound 1-IX (compound 1-101) or compound 1-IX A Compound 1-105 can be obtained. If the X group contains a moiety that can react under amide-forming conditions, a protected X' group (e.g., a protected terminal amine) may be used, which can then be deprotected and modified to reach the desired final compound (e.g., the terminal amine can be deprotected and then react with a suitable ester or acid to form the final amide with respect to X). Scheme 4 [ka]

[0228] Example 3: Synthesis of Compound 1-102 Compounds 1-XXII and 1-XXV, which are intermediates for many of the compounds 1-101 to 1-172, can be prepared as shown in Scheme 5. Precursor compounds 1-XXXII and 1-102 can be synthesized as shown in Scheme 6. The remaining compounds can be synthesized in a similar manner. Scheme 5 [ka] Scheme 6 [ka]

[0229] Example 4: General method for screening ARCS The ARCSs of this disclosure can be synthesized by those skilled in the art using common chemical synthesis principles and techniques. Alternatively, ARCSs can be prepared using known combinatorial methods to create large libraries of ARCSs. The ARCSs of this disclosure can also be synthesized as shown in Examples 1-3. ARCSs that covalently bind to biological targets on target cells can then be screened by gel assay, Western blot, ELISA, antibody array, or NanoBRET assay.

[0230] Example 5: Transfection protocol and reading for NanoBRET screening of ARCS Human embryonic kidney 293-H (HEK 293, Gibco 293-H, #11631017) cell lines are maintained in a water-saturated incubator at 37°C and 5% CO2 in Dulbecco's modified Eagle medium, high glucose, pyruvate (DMEM, Gibco, #11995065) supplemented with 10% fetal bovine serum (FBS, Gibco, #10082147) and 1× penicillin-streptomycin (100× solution, Gibco, #15140148). Cells are trypsinized using 0.05% or 0.25% trypsin-EDTA solution (trypsin-EDTA, phenol red, Gibco, #25200056 (0.25%) or #25300054). Opti-MEM medium supplemented with 10% fetal bovine serum (Opti-MEM I reduced serum medium, without phenol red, Gibco, #11058021) is used to culture cells overnight for NanoBRET reading experiments.

[0231] HEK293 cells are properly cultured before assay. The medium is removed from the cell flask by aspiration, washed once with PBS, then aspiration, trypsinized, and the cells are dissociated from the flask. The trypsin is neutralized using growth medium, and the cells are pelleted by centrifugation at 200 × g for 5 minutes. The medium is aspirated, and the cells are resuspended in a single cell suspension using Opti-MEM I supplemented with 10% FBS. The cell density is 2 × 10⁶ in Opti-MEM I supplemented with 10% FBS in a sterile conical tube. 5 The solution is adjusted to / mL. The cells are transfected and, the following day, directly aliquoted into 96-well plates for the NanoBRET assay, thus allowing the cells to be cultured overnight in Opti-MEM. The cells are also transfected in large quantities and dispensed into 96-well plates, allowing the cells to adhere to the plates overnight, thereby enabling the wash test.

[0232] Lipid:DNA complexes are prepared as follows:

[0233] A 10 μg / mL solution of DNA is prepared in serum-free Opti-MEM. This solution contains the carrier DNA encoding NanoLuc fused to the following biological targets in the following ratios. A serial dilution step may be ensured to accurately dilute the NanoLuc fusion DNA. In order, the following reagents were added to a sterile polystyrene test tube: 1 mL of Opti-MEM without phenol red; 9.0 μg / mL of carrier DNA; 1.0 μg / mL of NanoLuc fusion DNA (a smaller amount for some targets). The reagents were thoroughly mixed.

[0234] Add 30 μL of FuGENE HD to each mL of DNA mixture to form a lipid:DNA complex. Carefully pipette FuGENE HD directly into the liquid in the tube, ensuring that it does not touch the plastic side of the tube. Mix 5-10 times by pipetting up and down, and incubate at room temperature for 20 minutes to form the complex. Take a portion of the lipid:DNA complex (e.g., 1 mL) and divide it into 2 × 10⁻¹⁶ units. 5Mix 20 parts (e.g., 20 mL) of HEK293 cells in suspension with the ratio of 1 / mL and gently mix by pipetting up and down 5 times in a sterile conical tube. Larger or smaller bulk transfections are scaled accordingly using this ratio. Dispense 100 μL of cell + lipid:DNA complex into a sterile, tissue-culture-treated 96-well plate (20,000 cells / well) and incubate for at least 16 hours to express. Incubate cells in a 37°C + 5% CO2 incubator for more than 16 hours. Prepare serially diluted inhibitor or test compound at 100 × final concentration in 100% DMSO. Serially diluted inhibitor stocks are prepared on PCR plates. 100 serially diluted inhibitor / test compound Add 1 μL of the test compound per well to cells in a 96-well plate transfected transiently overnight, and mix by gently tapping the plate by hand. Incubate the plate overnight in a 37°C + 5% CO2 incubator. Prepare the appropriate concentrations of the substrate mixture (500X stock) and tracer in Opti-Mem. Wash the cells by setting a plate washer in a 96-well plate 5X in PBS at pH 7.4, adding 200 μL of PBS each time. Incubate the cells at 37°C for 2 hours. Add 100 μL of 1X Substrate-Tracer solution and mix by gently tapping the 96-well plate. Read the plate hourly on a plate reader for the next 6 hours. Several ARCS binding assays are shown below. Compound 1-XXXII formed covalent bonds with approximately 5%–20% of PI3 kinase. ARCS compounds selected from the group consisting of compounds 1-101, 1-102, 1-113, 1-114, 1-119, 1-120, 1-125, and 1-126 formed covalent bonds with approximately 80% to 100% of PI3 kinases. Compounds 1-171 and 1-172 formed covalent bonds with approximately 50% to 80% of PI3 kinases. PI3 kinase activity was inhibited by compound 1-XXXII by approximately 5% to 20%. PI3 kinase activity was inhibited by ARCS compounds selected from the group consisting of compounds 1-101, 1-102, 1-113, 1-114, 1-119, 1-120, 1-125, and 1-126 by approximately 80% to 100%. PI3 kinase activity was inhibited by compounds 1-171 and 1-172 by approximately 50% to 80%. [Table 1]

[0235] Equivalents and range Those skilled in the art can identify or confirm many equivalents to specific embodiments using only conventional experiments in accordance with the disclosures described herein. The scope of this disclosure is not intended to be limited to the above description, but is set forth in the appended claims.

[0236] In the claims, unless otherwise indicated or evident from the context, articles such as “a,” “an,” and “the” may mean one or more. A claim or description containing “or” among one or more members of a group is deemed satisfied if one, one or more, or all members of the group are present in, used in, or otherwise related to a given product or method, unless otherwise evident from the context. This disclosure includes embodiments in which exactly one member of the group is present in, used in, or otherwise related to a given product or method. This disclosure includes embodiments in which one or more, or all, members of the group are present in, used in, or otherwise related to a given product or method.

[0237] Furthermore, note that the term “contains” is open and permissive, but does not require the inclusion of any additional elements or steps. Wherever the term “contains” is used herein, the term “consisting of” is also encompassed and disclosed.

[0238] Where a range is given, it includes the endpoint. Furthermore, unless otherwise suggested or otherwise evident from the context and understanding of those skilled in the art, values ​​expressed as a range may be assumed to be any specific value or range within the ranges described in the different embodiments of this disclosure, up to one-tenth of the lower limit of the range, unless the context clearly indicates otherwise.

[0239] Furthermore, naturally, any particular embodiment of the Disclosure that belongs to the prior art may be expressly excluded from any one or more of the claims. Such embodiments may be excluded even if the exclusion is not expressly stated herein, as they are considered to be known to those skilled in the art. Any particular embodiment of the compositions of the Disclosure (e.g., any antibiotic, therapeutic agent or active ingredient, any manufacturing method, any method of use, etc.) may be excluded from any one or more of the claims for any reason, whether or not it relates to the existence of the prior art.

[0240] The terms used are descriptive rather than restrictive, and it should be understood that they may be modified within the scope of the appended claims without departing from the true scope and spirit of this disclosure in its broader form.

[0241] While this disclosure has been described in some length and with some specificity with respect to some of the described embodiments, it is not intended to be limited to such details or embodiments or any particular embodiment, but should be interpreted by reference to the appended claims in order to provide the broadest interpretation of such claims from the perspective of the prior art, and thus to effectively encompass the intended scope of this disclosure.

Claims

1. A therapeutic conjugate that forms a covalent bond with a kinase or pseudokinase.

2. The therapeutic conjugate according to claim 1, wherein the kinase is PI3 kinase (PI3K).

3. The aforementioned therapeutic conjugate, It has the structure (FCB)a-(L)b-(CLM)c, a and c are independent integers between 1 and 5. b is an integer between 0 and 5, The therapeutic conjugate according to claim 1 or claim 2, wherein the FCB portion comprises a PI3K inhibitor or a fragment, analog, or derivative thereof.

4. The aforementioned FCB, 【Chemistry 1】 A therapeutic conjugate according to claim 3, including the above.

5. The therapeutic conjugate according to claim 4, wherein the therapeutic conjugate comprises a structure selected from the group consisting of compounds 1-101 to 1-172.

6. structure 【Chemistry 2】 A therapeutic conjugate according to claim 1 or claim 2, having either or a pharmaceutically acceptable salt thereof, wherein L is 【Transformation 3】 Selected from the group consisting of, either end can be connected to the CLM, and R1 is, 【Chemistry 4】 Selected from the group consisting of, CLM is, 【Transformation 5】 A therapeutic conjugate selected from the group consisting of the following.

7. Compounds 1-101, 1-102, 1-103, 1-104, 1-105, 1-106, A therapeutic conjugate according to claim 6, selected from the group consisting of 1-107, 1-108, 1-109, 1-110, 1-111, 1-112, 1-113, 1-114, 1-115, 1-116, 1-117, 1-118, 1-119, 1-120, 1-121, 1-122, 1-123, 1-124, 1-125, 1-126, 1-127, 1-128, 1-129, 1-130, 1-137, 1-138, 1-145, 1-146, 1-153, 1-154, 1-161, 1-162, 1-169, 1-170, 1-171, and 1-172.

8. structure 【Transformation 6】 A therapeutic conjugate according to claim 1, having either or a pharmaceutically acceptable salt thereof, wherein L is 【Transformation 7】 And R2 is, 【Transformation 8】 Selected from the group consisting of, CLM is, 【Chemistry 9】 A therapeutic conjugate selected from the group consisting of the following.

9. A therapeutic conjugate according to claim 8, selected from the group consisting of compounds 1-131, 1-132, 1-133, 1-134, 1-135, 1-136, 1-139, 1-140, 1-141, 1-142, 1-143, 1-144, 1-147, 1-148, 1-149, 1-150, 1-151, 1-152, 1-155, 1-156, 1-157, 1-158, 1-159, 1-160, 1-163, 1-164, 1-165, 1-166, 1-167, and 1-168.

10. A therapeutic conjugate comprising a structure selected from compounds 1-101 to 1-172 or a pharmaceutically acceptable salt thereof.

11. A therapeutic conjugate comprising a structure selected from compounds 1-1 to 1-11 or a pharmaceutically acceptable salt thereof.

12. A pharmaceutical composition comprising a therapeutic conjugate according to any one of claims 1 to 11 and at least one pharmaceutically acceptable excipient.

13. A method for regulating the activity of a kinase or pseudokinase, comprising administering a therapeutic conjugate according to any one of claims 1 to 11.

14. The method according to claim 13, wherein the activity of the kinase or pseudokinase is inhibited.

15. The method according to claim 13, wherein the kinase is PI3K.

16. A method for treating a subject in need of treatment, comprising administering a therapeutically effective amount of the pharmaceutical composition described in claim 12.

17. The method according to claim 16, wherein the subject has a treatment condition selected from the group consisting of cancer, neurodegenerative disease, autoimmune disorder, and aging.

18. The method according to claim 17, wherein the subject has cancer.

19. The method according to claim 18, wherein the subject has cancer having a mutation in the PIK3CA gene.

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