PSMA-binding ligand-linker complex and method of use thereof

Abstract

This invention relates to small molecules having high affinity and specificity for prostate-specific membrane antigen (PSMA), and to methods for using them for therapeutic and diagnostic purposes.

Description

[Background technology]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Application No. 63 / 519,469, filed on 14 August 2023, and U.S. Provisional Application No. 63 / 520,054, filed on 16 August 2023, the disclosures of which are incorporated herein by reference in their entirety. [Technical Field]

[0002] This invention relates to small molecules having high affinity and specificity for prostate-specific membrane antigen (PSMA), and to methods for using them for therapeutic and diagnostic purposes. Related Technology Description

[0003] Prostate cancer (PCa) is the most commonly diagnosed cancer in American men and is one of the leading causes of cancer-related death, second only to lung cancer. Generally, patients with localized disease can be treated with radical prostatectomy and / or radiotherapy. Patients with metastatic forms of PCa can be temporarily treated with androgen deprivation strategies, but most patients with metastatic disease eventually experience disease progression and may progress to metastatic castration-resistant prostate cancer (mCRPC), which has a low survival rate. Chemotherapy (e.g., docetaxel, cabazitaxel) is often used after failure of anti-androgen therapy, but the response is often limited and is accompanied by associated on-target and off-target toxicities, particularly pronounced in older men. There is a clear need for better treatment approaches for patients with advanced PCa.

[0004] Prostate-specific membrane antigen (PSMA) is a type II transmembrane protein that is highly overexpressed in most prostate cancers. PSMA expression is further upregulated in poorly differentiated cancers, metastatic cancers, hormone-resistant cancers, and cancer cells in mCRPC patients. Furthermore, PSMA exhibits robust internal migration from the cell surface, making it an ideal target for imaging and therapy. In fact, radiopharmaceutical therapies targeting PSMA have shown promise in clinical trials, and several radioligand therapies and antibody-drug conjugates (ADCs) have demonstrated efficacy in preclinical trials. Relapse, off-target toxicity, issues related to immunogenicity of antibody-based therapies, and stringent requirements imposed on facilities manufacturing and managing radiopharmaceuticals pose significant challenges to the development, FDA approval, and use of drugs for treating mCRPC in the oncology community.

[0005] Therefore, there is still a need for an effective treatment that precisely localizes to prostate cancer lesions without significant off-target toxicity, is low in manufacturing costs, and avoids the regulatory challenges associated with the manufacture and handling of radiopharmaceuticals. [Overview of the project]

[0006] Small molecule-drug conjugates (SMDCs) offer an attractive alternative to the more conventional ADC approach for the delivery of PSMA-targeted chemotherapy agents. Both technologies typically contain a PSMA-targeting motif (antibody versus enzyme inhibitor), a spacer, a cleavageable linker, and a potent cytotoxic payload. These drugs are expected to bind to PSMA on the cell surface, then internalize and accumulate in endosomes and lysosomes, allowing for efficient release of the cytotoxic payload to target cells, usually by enzymatic degradation. However, compared to anti-PSMA antibodies, small molecule PSMA inhibitors exhibit similar precise localization to prostate cancer lesions, but their significantly lower molecular weight and simpler molecular properties result in lower manufacturing costs, greater flexibility in determining optimal dosing regimens, and higher tolerable doses. The toxicity profile of PSMA-SMDCs is expected to be significantly lower than that of ADCs. This is because they have shorter residence times and more rapid and uniform diffusion into tumor masses compared to normal organs such as the kidneys, lacrimal glands, and salivary glands.

[0007] Therefore, this specification provides therapeutic and diagnostic agents for prostate cancer utilizing the efficacy and specific affinity of small molecule inhibitors for PSMA. Accordingly, in one embodiment, this disclosure relates to a compound of structural formula (I). [ka] or a pharmaceutically acceptable salt thereof, in the formula, D, -NR 2 A therapeutic or diagnostic agent bound to L via a -, -S-, or -O- moiety, L is an acid-cleavable linker, which combines to provide a carbamate moiety in formula (I). Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R 2 It forms an amide with nitrogen having the following properties: m is an integer between 1 and 5. n is an integer from 1 to 4, and R 1 , R 2 , R 3 , and R4 The present invention provides compound (I) or a pharmaceutically acceptable salt thereof, each independently being H, a C1-C6 alkyl group, or a protecting group.

[0008] This disclosure also provides pharmaceutical compositions comprising the compounds disclosed herein and pharmaceutically acceptable excipients, carriers, adjuvants, stabilizers, and / or diluents.

[0009] One aspect of this disclosure provides a method for delivering a therapeutic or diagnostic agent to a subject. Such a method involves administering an effective therapeutic amount of a compound or pharmaceutical composition disclosed herein to a subject in need of such agent.

[0010] One aspect of this disclosure provides a method for treating a patient having cancer (for example, prostate cancer). Such a method involves administering to the patient an effective amount of the compounds or pharmaceutical compositions disclosed herein.

[0011] Another aspect of the present disclosure provides a method for imaging one or more cancer cells (such as prostate cancer cells) in a patient. Such a method involves administering an effective amount of the compounds or pharmaceutical compositions disclosed herein to the patient.

[0012] Another aspect of this disclosure relates to a compound of formula (II). [ka] or a pharmaceutically acceptable salt thereof, in the formula, Each Y is independently a natural or non-natural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety. m is an integer between 1 and 5, and The present invention provides a compound or a pharmaceutically acceptable salt thereof, where n is an integer between 1 and 4.

[0013] Another aspect of the present disclosure provides a compound of formula (III) Chemical Formula or a pharmaceutically acceptable salt thereof, wherein in the formula:[EOS] each X is independently a natural or non-natural n α-amino acid, wherein the C-terminus of X forms an amide with the nitrogen bearing R 2 , m is an integer from 1 to 5, n is an integer from 1 to 4, and R 1 , R 2 , R 3 , and R 4 are each independently H, C1-C6 alkyl, or a protecting group, provides the compound (I) or a pharmaceutically acceptable salt thereof.

[0014] Another aspect of the present disclosure provides a compound of formula (IV) Chemical Formula or a pharmaceutically acceptable salt thereof, wherein in the formula:[EOS] D is a therapeutic or diagnostic agent bound to L via a -NR 2 -, -S-, or -O- moiety, L is an acid-cleavable linker, which is bound to form a carbamate moiety in formula (IV), each Y is independently a natural or non-natural n α-amino acid, wherein the N-terminus of Y is bound to an adjacent carbonyl to provide an amide moiety, m is an integer from 1 to 5, and n is an integer from 1 to 4, provides the compound or a pharmaceutically acceptable salt thereof.

[0015] In another aspect, the present disclosure provides a method of synthesizing a compound of formula (I), the method comprising a compound of formula (V) Chemical Formula or a pharmaceutically acceptable salt thereof Compounds of formula (III) as described herein [ka] or a pharmaceutically acceptable salt thereof, in the formula, Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R 2 It forms an amide with nitrogen having the following properties: m is an integer between 1 and 5. n is an integer from 1 to 4, and R 1 , R 2 , R 3 , and R 4 However, each independently provides compound (I) or a pharmaceutically acceptable salt thereof, wherein each is H, a C1-C6 alkyl group, or a protecting group.

[0016] In another embodiment, the present disclosure relates to a method for synthesizing a compound of formula (IV), wherein the compound of formula (V) [ka] or a pharmaceutically acceptable salt thereof Compounds of formula (II) as described herein [ka] or a method comprising contacting with a pharmaceutically acceptable salt thereof, wherein the formula is: Each Y is independently a natural or unnatural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety. m is an integer between 1 and 5, and This provides a method where n is an integer between 1 and 4. [Brief explanation of the drawing]

[0017] The accompanying drawings are included to provide a further understanding of the compositions and methods of this disclosure, are incorporated herein, and constitute part of this specification. These drawings illustrate one or more embodiments of this disclosure and, together with the description, help to illustrate the principles and operation of this disclosure. [Figure 1A] This is the 1H NMR spectrum of compound 1 (CTT2274). [Figure 1B] This is the 31P NMR spectrum of compound 1 (CTT2274). [Figure 2A] This is a graph of the IC50 curves for compound 1 (CTT2274) and human PSMA. [Figure 2B] This is a graph of the IC50 curves for compound 1 (CTT2274) and mouse PSMA. [Figure 2C] This is a graph of the IC50 curves for compound 8 (CTT2101) and human PSMA. [Figure 3] Western blots of PSMA expression in PC3, PC3-PIP, and C4-2B cell lines, as well as in PDX tumors. [Figure 4A] This graph shows the in vitro efficacy of compound 8 (CTT2101), compound 1 (CTT2274), compound 2 (CTT227X), and free MMAE in PC3 cells. [Figure 4B] This graph shows the in vitro efficacy of compound 8 (CTT2101), compound 1 (CTT2274), compound 2 (CTT227X), and free MMAE in PC3-PIP cells. [Figure 4C] This graph shows the in vitro efficacy of compound 8 (CTT2101), compound 1 (CTT2274), compound 2 (CTT227X), and free MMAE in C4-2B cells. [Figure 5A] This graph shows the time-dependent changes in mouse body weight when comparing compound 8 (CTT2101) with MMAE. [Figure 5B] This graph shows the temporal multiplicative change in tumor volume when comparing compound 8 (CTT2101) with MMAE. [Figure 5C]This is a spaghetti plot showing the time course of mouse tumor volume for compound 8 (CTT2101). Each line represents one mouse. [Figure 5D] This is a spaghetti plot showing the time course of mouse tumor volume for MMAE. Each line represents one mouse. [Figure 5E] This is a spaghetti plot showing the time course of mouse tumor volume for PBS. Each line represents one mouse. [Figure 5F] These are the Kaplan-Meier survival curves for compound 8 (CTT2101), MMAE, and PBS. [Figure 5G] This is a Kaplan-Meier survival sub-analysis for compound 8 (CTT2101) based on initial tumor volume. [Figure 6A] This graph shows the relationship between tumor volume and the number of days of treatment in mice treated with compound 8 (CTT2101). [Figure 6B] This graph shows the relationship between tumor volume and the number of days of treatment in mice treated with compound 8 (CTT2101). [Figure 6C] This graph shows the relationship between tumor volume and the number of days of treatment in mice treated with MMAE. [Figure 6D] This graph shows the relationship between tumor volume and the number of days of treatment in mice treated with MMAE. [Figure 7A] This graph shows the time course of mouse body weight for compound 8 (CTT2101), compound 1 (CTT2274), MMAE, and PBS. [Figure 7B] This graph shows the temporal multiplicative changes in tumor volume for compound 8 (CTT2101), compound 1 (CTT2274), MMAE, and PBS. [Figure 7C] This is a spaghetti plot showing the time course of mouse tumor volume for compound 8 (CTT2101). [Figure 7D] This is a spaghetti plot showing the time course of mouse tumor volume for compound 1 (CTT2274). [Figure 7E]This is a spaghetti plot showing the temporal changes in mouse tumor volume for MMAE. [Figure 7F] This is a spaghetti plot showing the temporal changes in mouse tumor volume for PBS. [Figure 7G] These are Kaplan-Meier survival curves for compound 8 (CTT2101), compound 1 (CTT2274), MMAE, and PBS. [Figure 8] This is a plot of body weight and blood concentration of the analyte in mice treated with compound 1 (CTT2274) and MMAE. [Figure 9A] This is a spaghetti plot showing the time course of mouse body weight for compound 1 (CTT2274). [Figure 9B] This is a spaghetti plot showing the time course of tumor volume for compound 1 (CTT2274). [Figure 9C] This is a spaghetti plot showing the temporal multiplicative change in tumor volume for compound 1 (CTT2274). [Figure 9D] This is the Kaplan-Meier survival curve for compound 1 (CTT2274). [Figure 10] This is a spaghetti plot showing the time course of tumor volume in individual mice for compound 1 (CTT2274). [Modes for carrying out the invention]

[0018] Before the disclosed processes and materials are described, it should be understood that the embodiments described herein are not limited to any particular embodiment and are therefore naturally diverse. It should also be understood that the terms used herein are intended solely to describe a particular embodiment and are not intended to be limiting unless specifically defined herein.

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art in which the present invention pertains. All patents, patent applications, and publications referenced herein are incorporated by reference to the extent that they correspond to this disclosure. Unless otherwise expressly defined, terms and scopes have their generally defined meanings.

[0020] For brevity, the chemical moieties are defined and referred to throughout as primarily monovalent chemical moieties (e.g., alkyl, aryl, etc.). Nevertheless, such terminology may also be used to convey the corresponding multivalent moieties under appropriate structural circumstances that are obvious to those skilled in the art. For example, while the “alkyl” moiety generally refers to a monovalent radical (e.g., CH3-CH2-), in certain circumstances a divalent linkage moiety can be “alkyl,” in which case those skilled in the art will understand alkyl to be a divalent radical (e.g., -CH2-CH2-) equivalent to the term “alkylene.” Similarly, in situations where a divalent moiety is required and it is stated that it is “aryl,” those skilled in the art will understand that the term “aryl” refers to the corresponding divalent moiety, arylene. It is understood that all atoms have their usual valence numbers for bond formation (i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S depending on the oxidation state of S).

[0021] The term "amino" refers to -NH2.

[0022] The term "acetyl" refers to -C(O)CH3.

[0023] As used herein, the term "acetyl" refers to an alkylcarbonyl or arylcarbonyl substituent, where the alkyl and aryl portions are as defined herein.

[0024] As used herein, the term "alkyl" refers to saturated linear and branched aliphatic groups having 1 to 12 carbon atoms. Therefore, "alkyl" refers to C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 It contains a group. The alkyl group may be branched or unbranched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0025] As used herein, the term “alkenyl” means an unsaturated linear or branched aliphatic group having one or more carbon-carbon double bonds with 2 to 12 carbon atoms. Thus, “alkenyl” refers to C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 It includes alkenyl groups. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0026] As used herein, the term "alkynyl" means an unsaturated linear or branched aliphatic group containing one or more carbon-carbon triple bonds having 2 to 12 carbon atoms. Thus, "alkynyl" refers to C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 It contains the group. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0027] An "alkylene," "alkenylene," or "alkynylene" group is an alkyl, alkenyl, or alkynyl group that is located between two other chemical groups and functions to link them, as defined herein. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, and butylene. Examples of alkenylene groups include, but are not limited to, ethenylene, propenylene, and butenylene. Examples of alkynylene groups include, but are not limited to, etynylene, propynylene, and butynylene.

[0028] The term "alkoxy" refers to -O (C1-C6 alkyl).

[0029] As used herein, the term "cycloalkyl" refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbon atoms. Therefore, "cycloalkyl" includes C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 Examples include cyclic hydrocarbon groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. The "cycloalkyl" group also includes condensed polycyclic (e.g., bicyclic) ring systems such as decahydronaphthyl and octahydro-1H-indenyl. One of the condensed rings may be non-aromatic, provided that at least one of the rings is aromatic, for example, indenyl.

[0030] The term "C3-C6 cycloalkyloxy" refers to a group with the formula -O(C3-C6 cycloalkyl).

[0031] The "aryl" group is a C6-C group containing 1 to 3 aromatic rings. 14 This is the aromatic part. Therefore, as an "aryl" group, it is C6, C 10 , C 13 , and C14 Examples include cyclic hydrocarbon groups. Typical aryl groups include C6-C 10 The group is an aryl group. Specific aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and fluorenyl. The "aryl" group also includes fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings are non-aromatic, provided that at least one ring is aromatic, such as an indenyl ring.

[0032] An "aralkyl" or "arylalkyl" group contains an aryl group covalently bonded to an alkyl group, with this portion linked to another group via the alkyl moiety. Typical aralkyl groups include, but are not limited to, benzyl, phenethyl, and naphthylmethyl -(C1-C6)alkyl(C6-C 10 ) is an aryl group. For example, arC1-C3 alkyl is an aryl group covalently bonded to a C1-C3 alkyl group.

[0033] As used herein, the term “condensation” refers to a ring system (including bridged ring systems) that shares two or more atoms, such as a bicyclic or tricyclic ring system, for example, when used to define a bicyclic condensed ring system. Examples of such condensed ring systems include (1S,4R)-2-azabicyclo[2.2.1]heptane, 2-azabicyclo[2.2.2]octane, 2,5-diazabicyclo[2.2.2]octane, 2-oxa-5-azabicyclo[2.2.2]octane, isoindoline, 1,2,3,4-tetrahydro-2,6-naphthyridine, 1,2,3,4-tetrahydroisoquinoline, 1,2,3,4-tetrahydro-1,4-(epiminomethano)naphthalene, and 3-azabicyclo[3.1.0]hexane.

[0034] A "heterocyclyl," "heterocyclic," or "heterocycloalkyl" group is a monocyclic or bicyclic (e.g., fused) ring structure having 3 to 12 atoms (3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 atoms), for example, 4 to 8 atoms, where one or more ring atoms are independently N, O, or S, and the remaining ring atoms are quaternary or carbonyl carbons. Examples of heterocyclic groups include, but are not limited to, epoxy, oxylanil, oxetanil, azetidinil, azilidinil, tetrahydrofuranil, tetrahydropyranil, tetrahydrothiophenyl, pyrrolidinil, piperidinil, piperazinil, imidazolidinil, thiazolidinil, thiatanil, dithianil, trithianil, azathianil, oxathianil, dioxolanil, oxazolidinil, oxazolidinil, decahydroquinolinil, piperidonil, 4-piperidonil, thiomorpholinil, dimethylmorpholinil, and morpholinil. Compounds having adjacent ring O and / or S atoms are particularly excluded from the scope of this term. Heterocyclic groups can be bonded to a parent group (i.e., a bonding site) via any ring atom containing one heteroatom or one carbon atom in the heterocyclic group. If chemically necessary, the heterocycle may be bonded to one or more other groups, for example, when acting as a bridging group. The term “heterocyclyl” also includes fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings are aromatic or non-aromatic, provided that at least one ring is non-aromatic and contains an N, O, or S ring atom. Examples of such fused polycyclic ring systems include indolinyl, indolin-2-yl, 2,3-dihydrobenzofuran-2-yl, and 2,3,4,5-tetrahydrobenzo[d]oxazole-2-yl. Each of these examples is a nine-membered heterocyclyl.

[0035] As used herein, the term “heteroaryl” refers to a group having 5 to 14 ring atoms, preferably 5, 6, 10, 13, or 14 ring atoms, and having 6, 10, or 14 π electrons shared in the cyclic array, and having 1 to 3 heteroatoms, each independently being N, O, or S, in addition to the carbon atoms. “Heteroaryl” also includes fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings are non-aromatic, provided that at least one ring is aromatic and at least one ring contains an N, O, or S ring atom. A heteroaryl group can be bonded to a parent group (i.e., a bonding site) via any ring atom containing one of the heteroatoms or one of the carbon atoms in the heteroaryl group. If chemically required, a heteroaryl may be bonded to one or more other groups, for example, when acting as a bridging group.

[0036] Examples of heteroaryl groups include acridinyl, azosinyl, benzimidazolyl, benzofuranil, benzo[d]oxazole-2(3H)-one, 2H-benzo[b][1,4]oxazine-3(4H)-one, benzothiofuranil, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanil, chromenyl, sinnolinyl, furanil, and furanil. Lazanil, imidazolinil, imidazolyl, 1H-indazolyl, indolenil, indolinyl, indolidinyl, indolyl, 3H-indolyl, isobenzofuranil, isochromanil, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinil, isothiazolyl, isoxazolyl, naphthilidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazo Ryl, oxazolidinil, pyrimidinil, phenanthrolinil, phenanthrolinil, phenazinil, phenothiazinil, phenoxathiinil, phenoxazinil, phthalazinil, piperonil, pteridinil, purinil, pyranil, pyrazinil, pyrazolidinil, pyrazolil, pyrazolyl, pyridazinil, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinil, pyridyl, pyrimidinil, pyrrolinil, 2H-pyrrolyl, pyrrrolyl, quinazolinil, quinolinil, 4H-quinolidinil, quinoxalinil, quinucrine Examples include dinyl, 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, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl.

[0037] An "arylene," "heteroarylene," or "heterocyclylene" group is a divalent aryl, heteroaryl, or heterocyclyl group, as defined herein, that is located between two other chemical groups and serves to connect them.

[0038] As used herein, when a moiety (e.g., cycloalkyl, aryl, heteroaryl, heterocyclyl, urea, etc.) is described as "optionally substituted" without explicitly specifying the substituents, it means that the group optionally has multiple nonhydrogen substituents, for example, 1 to 5, 1 to 4, 1 to 3, 1, or 2 nonhydrogen substituents.

[0039] As used herein, the terms "halogen" or "halo" refer to chlorine, bromine, fluorine, or iodine.

[0040] The term "haloalkyl" refers to an alkyl chain in which one or more hydrogen atoms are replaced by halogens. Typical haloalkyls include trifluoromethyl, difluoromethyl, fluorochloromethyl, chloromethyl, and fluoromethyl.

[0041] The term "hydroxyalkyl" refers to -alkylene-OH.

[0042] As used herein, the term “substituted” means that a hydrogen radical in a specified part is replaced by a radical of a particular substituent, provided that such substitution results in a stable or chemically feasible compound. The term “substitutable,” when used in relation to a specified atom, means that a hydrogen radical is bonded to that atom and that the hydrogen radical is substituted by a radical of a preferred substituent.

[0043] As used herein, the term “one or more” substituents means one or more substituents up to the maximum number of substituents possible based on the number of available bonding sites, provided that the stability and chemical feasibility conditions are met. Unless otherwise specified, optionally substituted groups may have substituents at each of their substitutable positions, and the substituents may be identical or different. As used herein, the term “independently selected” means that for multiple instances of a given variable in a compound, the same or different values ​​may be selected.

[0044] As used herein, “protecting group” includes, but is not limited to, optionally substituted benzyl, t-butyl esters, allyl esters, alkyl esters (e.g., methyl, ethyl), fluorenyl methoxycarbonyl groups (Fmoc), and amino, carboxylic acid, and phosphate protecting groups as described in Greene's Protective Groups in Organic Synthesis, 4th Edition (which is incorporated by reference). In some embodiments, R 1 is a carboxylic acid protecting group (e.g., methyl or t-butyl ester). In some embodiments, R 2 The group is a nitrogen protecting group (e.g., Boc, or benzyl). Optionally, examples of benzyl groups include, but are not limited to, unsubstituted benzyl, triphenylmethyl (trityl), diphenylmethyl, o-nitrobenzyl, 2,4,6-trimethylbenzyl, p-bromobenzyl, p-nitrobenzyl, p-methoxybenzyl (PMB), 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl, 4-sulfobenzyl, 4-azidomethoxybenzyl, and piperonyl, as well as the benzyl protecting groups of carboxylic acids and phosphoric acids disclosed in Greene's Protective Groups in Organic Synthesis (the relevant portion of which is incorporated herein by reference).

[0045] The individual atoms in formula (I) and the compounds within formula (I) may exist in any of their naturally occurring isotopes, but it is understood that the most abundant isotope is preferred. Therefore, as an example, the individual hydrogen atoms in formula (I) or in the formulas shown below, 1 H, 2 H (deuterium; D), or 3 It may exist as a H (tritium; T) atom, preferably 1 It may exist as H. Similarly, as an example, each carbon atom present in formula (I), or the formulas shown below, 12 C, 13 C, or, 14 It may exist as a C atom, preferably 12 It may exist as C.

[0046] As used herein, an "effective amount" of the compound is sufficient to negatively modulate or inhibit the activity of PSMA.

[0047] As used herein, a “therapeutic dose” of a compound is an amount sufficient to alleviate or reduce symptoms in any way, to halt or halt the progression of a condition, or to negatively modulate or inhibit PSMA activity. Such a dose may be administered as a single dose or according to a regimen, thereby being effective.

[0048] As used herein, “treatment” means any form of treatment that alleviates or beneficially alters the symptoms or condition of a patient’s condition, disorder, or disease.

[0049] In consideration of this disclosure, the methods and compositions described herein can be configured by those skilled in the art to satisfy desired needs. Generally, the disclosed compositions and methods utilize the potency and specific affinity of PSMA small molecule inhibitors to bring about improvements in the treatment of cancer, particularly prostate cancer.

[0050] compound One aspect of this disclosure relates to a compound having structural formula (I). [ka] or a pharmaceutically acceptable salt thereof, in the formula, D, -NR 2 A therapeutic or diagnostic agent bound to L via a -, -S-, or -O- moiety, L is an acid-cleavable linker that combines to provide the amide moiety in formula (I), Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R 2 It forms an amide with nitrogen having the following properties: m is an integer between 1 and 5. n is an integer from 1 to 4, and R 1 , R 2 , R 3 , and R 4 However, each independently provides compound (I) or a pharmaceutically acceptable salt thereof, wherein each is H, a C1-C6 alkyl group, or a protecting group.

[0051] The compound of structural formula (I) may have a specific stereochemistry. For example, in a particular embodiment, the present disclosure provides a compound having the following formula: [ka] In the formula, D, L, X, m, n, R 1 , R 2 , R 3 , and R 4 The present specification provides a compound having the formula as described herein.

[0052] The compounds of the present disclosure described herein each comprise at least one X, where each X is independently a natural or non-natural n α-amino acid. As shown in formula (I), the C-terminus of X is R 2 It forms an amide with nitrogen having [a specific characteristic]. Furthermore, the N-terminus of X is bonded to an adjacent carbonyl to provide the amide moiety.

[0053] In certain embodiments, each X is independently selected from naturally occurring amino acids (e.g., L-amino acids) or unnaturally occurring amino acids (e.g., D-amino acids). In certain embodiments described herein, each X is independently selected from naturally occurring amino acids (e.g., L-amino acids). For example, in certain embodiments, at least one X is independently selected from alanine, glycine, isoleucine, leucine, proline, valine, phenylalanine, tryptophan, tyrosine, aspartic acid, glutamic acid, arginine, histidine, lysine, serine, threonine, cysteine, methionine, asparagine, or glutamine. In certain embodiments, at least one X is selected from phenylalanine, tryptophan, tyrosine, or histidine. In certain embodiments, at least one X is phenylalanine. In certain embodiments, at least one X is tryptophan. In certain embodiments, at least one X is 4-biphenylalanine. If X is a naturally occurring amino acid, the side chain of such an amino acid may be further substituted. For example, the amino acid side chain may be H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6alkyl), halogen, C3-C 10 Cycloalkyl, C3-C8 heterocyclyl, aryl, heteroaryl, C3-C 10 It may be substituted with a cycloalkyl (C1-C6 alkyl), a C3-C8 heterocyclyl (C1-C6 alkyl), an aryl (C1-C6 alkyl), or a heteroaryl (C1-C6 alkyl), where R 10 and R 11 However, each is independently selected from hydrogen and C1-C6 alkyl groups.

[0054] In certain embodiments described herein, at least one X has the following structure: [ka]

[0055] In a particular embodiment, each R 6 Each of these is independently (i) H, (ii) C1-C6 alkyl, (iii) hydroxy(C1-C6 alkyl), (iv) C1-C6 alkoxy(C1-C6 alkyl), (v) C2-C6 alkenyl, (vi) R 8 R 9 N(C1-C6 alkyl), (vii) halogen, (viii) halo(C1-C6 alkyl), (ix) carboxy(C1-C6 alkyl), (x) thio(C1-C6 alkyl), (xi) C1-C6 alkylthio(C1-C6 alkyl), (xii) R 8 R 9 NC(O)(C1-C6 alkyl), (xiii)C3-C 10 Cycloalkyl (C1-C6 alkyl), (xiv)C3-C8 heterocyclyl (C1-C6) alkyl, (xv) aryl (C1-C6 alkyl), (xvi) heteroaryl (C1-C6 alkyl), (xvii) R 17 C(O)NR 8 (C1-C6 alkyl), (xviii)C3-C 10 Selected from cycloalkyl, (xix)C3-C8 heterocyclyl, (xx)aryl, and (xvi) heteroaryl, where R 8 and R 9 R is independently selected from hydrogen and C1-C6 alkyl (e.g., C1-C4 alkyl or C1-C3 alkyl), and here R 17 However, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl), C3-C 10cycloalkyl(C1-C6 alkyl), C3-C8 heterocyclyl(C1-C6 alkyl), aryl(C1-C6 alkyl), heteroaryl(C1-C6 alkyl), C3-C 10 cycloalkyl, C3-C8 heterocyclyl, aryl, or heteroaryl. In certain specific embodiments, each cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl), halogen, C3-C 10 cycloalkyl(C1-C6 alkyl), C3-C8 heterocyclyl(C1-C6 alkyl), aryl(C1-C6 alkyl), or heteroaryl(C1-C6 alkyl), wherein R 10 and R 11 are each independently selected from hydrogen and C1-C6 alkyl (for example C1-C4 alkyl, or C1-C3 alkyl).

[0056] In certain specific embodiments described herein, at least one X has the structure shown below.

Chemical Formula

[0057] In certain specific embodiments, p is an integer of 1 to 5 (for example, 1 to 4, or 1 to 3). In certain specific embodiments, q is an integer of 1 to 5 (for example, 1 to 4, or 1 to 3). In certain specific embodiments, each R 7 is each independently H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11N(C₁-C₆ alkyl), halogen, C₃-C 10 cycloalkyl, C₃-C₈ heterocyclyl, aryl, heteroaryl, C₃-C 10 cycloalkyl(C₁-C₆ alkyl), C₃-C₈ heterocyclyl(C₁-C₆ alkyl), aryl(C₁-C₆ alkyl), and heteroaryl(C₁-C₆ alkyl), wherein each cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with -OH, -SH, -NO₂, -COOH, C₁-C₆ alkyl, hydroxy(C₁-C₆ alkyl), C₁-C₆ alkoxy(C₁-C₆ alkyl), C₂-C₆ alkenyl, -NR 10 R 11 , R 10 R 11 N(C₁-C₆ alkyl), halogen, C₃-C 10 cycloalkyl(C₁-C₆ alkyl), C₃-C₈ heterocyclyl(C₁-C₆ alkyl), aryl(C₁-C₆ alkyl), or heteroaryl(C₁-C₆ alkyl), and R 10 and R 11 are each independently selected from hydrogen and C₁-C₆ alkyl (e.g., C₁-C₄ alkyl, or C₁-C₃ alkyl). For example, in certain embodiments, p is 1. In some embodiments described herein, q is 1. In certain embodiments, p is 1, q is 1, and R 7 is aryl (e.g., phenyl).

[0058] Each R 17 is each independently selected from C₁-C₆ alkyl, hydroxy(C₁-C₆ alkyl), C₁-C₆ alkoxy(C₁-C₆ alkyl), C₂-C₆ alkenyl, -NR 10 R 11 , R 10 R 11 N(C₁-C₆ alkyl), C₃-C 10 cycloalkyl, C₃-C₈ heterocyclyl, aryl, heteroaryl, C₃-C 10Selected from cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), and heteroaryl (C1-C6 alkyl), where R 10 and R 11 These are independently selected from hydrogen and C1-C6 alkyl (e.g., C1-C4 alkyl, or C1-C3 alkyl).

[0059] In certain embodiments described herein, at least one X has the following structure: [ka]

[0060] In a particular embodiment, r is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, s is an integer between 1 and 7 (for example, 1 to 5, or 1 to 4, or 1 to 3). In a particular embodiment, each R 12 These are, independently, H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, and -NR. 10 R 11 , R 10 R 11 N(C1-C6 alkyl), halogen, C3-C 10 Cycloalkyl, C3-C8 heterocyclyl, aryl, heteroaryl, C3-C 10 The compounds are selected from cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), and heteroaryl (C1-C6 alkyl), where each cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, and -NR. 10 R 11 , R 10 R 11N(C1-C6 alkyl), halogen, C3-C 10 Substituted with cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), or heteroaryl (C1-C6 alkyl), R 10 and R 11 R is independently selected from hydrogen and C1-C6 alkyl (e.g., C1-C4 alkyl, or C1-C3 alkyl). For example, in a particular embodiment, r is 1. In some embodiments described herein, s is 1. In a particular embodiment, r is 1 and s is 1. In a particular embodiment, R 12 H is H.

[0061] In certain embodiments described herein, at least one X has the following structure: [ka]

[0062] In a particular embodiment, a is an integer between 1 and 4 (for example, 1 to 3). In a particular embodiment, b is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, R 13 and R 14 Each of these is independently H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl) and halogens are selected, where R 10 and R 11 However, each is independently selected from hydrogen and C1-C6 alkyl (for example, C1-C4 alkyl, or C1-C3 alkyl). In a particular embodiment, R 13 and R 14 H is H.

[0063] In certain embodiments described herein, at least one X has the following structure: [ka]

[0064] In a particular embodiment, c is an integer between 1 and 3 (for example, 1 or 2). In a particular embodiment, d is an integer between 1 and 4 (for example, an integer between 1 and 3). In a particular embodiment, R 15 and R 16 Each of these is independently H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl) and halogens are selected, where R 10 and R 11 However, each is independently selected from hydrogen and C1-C6 alkyl (for example, C1-C4 alkyl, or C1-C3 alkyl). In a particular embodiment, R 15 and R 16 H is H.

[0065] In certain embodiments described herein, at least one X has the following structure: [ka]

[0066] In a particular embodiment, t is an integer between 1 and 6 (for example, 1 to 5, or 1 to 4). In a particular embodiment, u is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, v is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, each R 18These are, independently, H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, and -NR. 10 R 11 , R 10 R 11 N(C1-C6 alkyl) and halogens are selected, where R 10 and R 11 However, each is independently selected from hydrogen and C1-C6 alkyl (for example, C1-C4 alkyl or C1-C3 alkyl). In a particular embodiment, each R 18 Each of these is independently selected from C1-C6 alkyl groups.

[0067] In certain embodiments described herein, at least one X has the following structure: [ka]

[0068] As described above, the compound of structural formula (I) is one of the above embodiments in which n is an integer from 1 to 4 or from 1 to 3. In a particular embodiment, n is 1 or 2. In other embodiments, n is 1. For example, in some embodiments, n is 1 and X is phenylalanine. In other embodiments, n is 1 and X is as follows: [ka] In other embodiments described herein, n is 1 and X is as follows: [ka]

[0069] In certain embodiments, the compound of formula (I) is one of the previously described embodiments where m is an integer from 1 to 5. For example, in certain embodiments described herein, m is an integer from 1 to 4, or from 1 to 3. In certain embodiments, m is 1 or 2. In other embodiments of the present disclosure described herein, m is 1.

[0070] In a particular embodiment, the compound is R 1 , R 2 , and R 3 However, independently, it is one of the preceding embodiments selected from one of bases (a) to (o). (a) Hydrogen, C1-C6 alkyl, or protecting group. (b) Hydrogen or C1-C6 alkyl. (c) C1-C6 alkyl or protecting group. (d) C1-C6 alkyl. (e) Hydrogen or protecting group. (f) Hydrogen. (g) Protecting groups. (h) Any of the groups (a) to (d) wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl. (i) Any of the groups (a) to (d) wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, or tert-butyl. (j) Any of the groups (a) to (d) in which the C1-C6 alkyl group is methyl, ethyl, n-propyl, or tert-butyl. (k) Any of the groups (a) to (d) in which the C1-C6 alkyl group is methyl, ethyl, or tert-butyl. (l) Any of the groups (a) to (d) in which the C1-C6 alkyl group is methyl or ethyl. (m) Any of the groups (a) to (d) in which the C1-C6 alkyl group is methyl. (n) Any of the groups (a) to (d) in which the C1-C6 alkyl group is ethyl. (o) Any of the groups (a) to (d) in which the C1-C6 alkyl group is tert-butyl.

[0071] In certain embodiments of the compounds of this disclosure described herein, R 1 , R 2 , and R 3 H is H.

[0072] In a particular embodiment, R 1 , R 2 , and R 3 At least one of them is not H, but R 1 , R 2 , and R 3 The remaining value is H.

[0073] In a particular embodiment, the compound is R 4 In one particular embodiment, R is either H or methyl. 4 H is H.

[0074] The compounds of this disclosure described herein include an acid-cleaving linker L (for example, a linker that degrades in an acidic environment or under other desired conditions to release a therapeutic or diagnostic agent). By binding such a linker to a drug, controlled release of the drug can be induced by taking advantage of the fact that the intracellular pH (e.g., pH 5.5) or extracellular pH (e.g., pH 6.5) of the target cell type is lower than that of blood or normal tissue.

[0075] As described above, L bondes to provide a carbamate moiety in formula (I). In certain embodiments of this disclosure, the linker L comprises a phosphoramidate group. For example, in certain embodiments, L comprises a phosphoramidate of the following formula: [ka] R 1 and R 3 This is as defined above with respect to formula (I). In a particular embodiment, R1 is hydrogen, and / or R 3 It is hydrogen.

[0076] In certain embodiments of this disclosure, L is selected from the following: [ka] Here, R 5 is H, -OH, or C1-C6 alkoxyl, and R 1 , R 2 , and R 3 This is as defined above with respect to equation (I).

[0077] In certain embodiments of this disclosure, L is selected from the following: [ka] Here, R 5 is H, -OH, or C1-C6 alkoxyl, and R 1 , R 2 , and R 3 This is as defined above with respect to equation (I).

[0078] In certain embodiments of this disclosure, R 5 is H, -OH, or C1-C4 alkoxyl. In some embodiments, R 5 is H, -OH, or C1-C2 alkoxyl. In some embodiments, R 5 is H, -OH, or -OCH3. In some embodiments, R 5 is H, or -OCH3. In some embodiments, R 5 is H. In some embodiments, R 5 It is -OCH3.

[0079] In certain embodiments of this disclosure, L is as follows: [ka]

[0080] As used herein, the term "PhosAm-2" is used to refer to a compound of formula (I) having such a linker L. "PhosAm-2"-containing compounds can be derived from "PhosAm-A"-containing azides of formula (V), as described below.

[0081] In certain embodiments of this disclosure, L is selected from the following: [ka] Here, R 1 , R 2 , and R 3 This is as defined above with respect to equation (I).

[0082] In certain embodiments of this disclosure, L is as follows: [ka]

[0083] As used herein, the term "PhosAm-1" is used to refer to a compound of formula (I) having such a linker L. "PhosAm-1"-containing compounds can be derived from "PhosAm-B"-containing azides of formula (V), as described below.

[0084] As specified above, the compounds of formula (I) described herein contain D, which is a therapeutic or diagnostic part. Generally, -NR 2 It is bound to linker L via -, -S-, or -O-. It should be understood that the drug can actually be a derivative modified at the linking site. For example, D is bound to L via -NR 2 It can be modified to consist of -, -S-, or -O-.

[0085] As described above, in some embodiments, D is a therapeutic agent. A therapeutic agent is a molecule useful for treating a disease. Examples of therapeutic agents include chemotherapeutic agents, antibodies, antibody fragments, toxins, enzymes, nucleases such as ribonucleases (RNases) and DNase I, hormones, cytokines, chemokines, angiogenesis inhibitors, antisense oligonucleotides, small interfering RNAs (siRNAs), chelating agents, boron compounds, photoactivators, small molecules, antibiotics, and radioisotopes. For example, in some embodiments described herein, D is an anti-angiogenic agent, a cytotoxic agent, a cytokine, a chemokine, an apoptotic agent, a prodrug, a toxin, an enzyme, a radioisotope, an immunomodulator, an antibiotic, a CNS-active agent, or a hormone.

[0086] Chemotherapy agents include, for example, anticancer agents, anti-cancer agents, and cytotoxic agents. Examples of anticancer chemotherapeutic agents include, but are not limited to, 5-fluorouracil, bleomycin, busulfan, camptothecin, carboplatin, chlorambucil, cisplatin (CDDP), cyclophosphamide, dactinomycin, daunorubicin, doxorubicin, estrogen receptor conjugates, etoposide (VP16), farnesyl-protein transferase inhibitors, gemcitabine, ifosfamide, mechloretamine, melphalan, methotrexate, mitomycin, navelbine, nitrosourea, plicamycin, procarbazine, raloxifene, tamoxifen, taxol, temazolomide (aqueous form of DTIC), transplatinum, vinblastine and methotrexate, vincristine, or variants of any of the above analogues or derivatives.Chemotherapy agents used against infectious organisms include acyclovir, albendazole, amantadine, amikacin, amoxicillin, amphotericin B, ampicillin, aztreonam, azithromycin, bacitracin, bactrim, batrafen, bifonazole, carbenicillin, caspofungin, cefaclor, cefazolin, cephalosporin, cefepime, ceftriaxone, cefotaxime, chloramphenicol, cidofovir, cypro(registered trademark), clarithromycin, clavulanic acid, clotrimazole, cloxacillin, doxycycline, econazole, erythrocycline, erythromycin, Flagyl(registered trademark), fluconazole, flucytosine, foscarnet(registered trademark), furazolidone, ganciclovir, and ge Examples include, but are not limited to, antamicin, imipenem, isoniazid, itraconazole, kanamycin, ketoconazole, lincomycin, linezolid, meropenem, miconazole, minocycline, naphthifine, nalidixic acid, neomycin, netylmycin, nitrofurantoin, nystatin, oseltamivir, oxacillin, paromomycin, penicillin, pentamidine, piperacillin-tazobactam, rifabutin, rifampin, rimantadine, streptomycin, sulfamethoxazole, sulfasalazine, tetracycline, thioconazole, tobramycin, tolcyclate, tolnaftate, trimethoprim-sulfamethoxazole, valacyclovir, vancomycin, zanamil, and zithromycin.

[0087] Hormones can be used as therapeutic agents, either on their own or in combination with other chemotherapy agents. Progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate have been used for endometrial and breast cancers. Estrogens such as diethylstilbestrol and ethinylestradiol have been used for cancers such as prostate cancer. Anti-estrogens such as tamoxifen have been used for cancers such as breast cancer. Androgens such as testosterone propionate and fluoxymesterone have also been used to treat breast cancer. Corticosteroid hormones such as prednisone and dexamethasone can enhance the effects of other chemotherapy agents. Cytokines used as therapeutic agents include, but are not limited to, lymphokines, monokines, growth factors, and polypeptide hormones. Examples of cytokines include human growth hormone, N-methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), luteinizing hormone (LH), hepatic growth factor, prostaglandins, fibroblast growth factor, prolactin, placental lactogen, OB protein, tumor necrosis factor-α, tumor necrosis factor-β, Müllerian duct inhibitors, mouse gonadotropin-related peptides, inhibin, activin, vascular endothelial growth factor, integrin, thrombopoietin (TPO), NGF-β, platelet growth factor, TGF-α, TGF-β, and insulin-like compounds. Examples include growth factor I, insulin-like growth factor II, erythropoietin (EPO), bone induction factor, interferon-α, interferon-β, interferon-γ, macrophage-CSF (M-CSF), granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (G-CSF), IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, LIF, kit-ligand, FLT-3, angiostatin, thrombospondin, endostatin, and lymphotoxin.Examples of angiogenesis inhibitors used as therapeutic agents include angiostatin, baculostatin, canstatin, maspin, anti-VEGF antibodies, anti-PIGF peptides and antibodies, anti-angiogenic growth factor antibodies, anti-Flk-1 antibodies, anti-Flt-1 antibodies and peptides, laminin peptides, fibronectin peptides, plasminogen activator inhibitors, tissue metalloproteinase inhibitors, interferon, interleukin-12, IP-10, Gro-β, thrombospondin, 2-methoxyestradiol, and p Examples include, but are not limited to, loriferin-related proteins, carboxamide triazole, CM101, marimast, pentosan polysulfate, angiopoietin-2, interferon-α, harbimycin A, PNU145156E, 16K prolactin fragment, linamide, thalidomide, pentoxifylline, genistein, TNP-470, endostatin, paclitaxel, acutine, angiostatin, cidofovir, vincristine, bleomycin, AGM-1470, platelet factor 4, and minocycline. Examples of small molecules for use as therapeutic agents include abrin, amantadine, amoxicillin, amphotericin B, ampicillin, apridin, azalibine, anastrozole, azacitidine, aztreonam, azithromycin, bacitracin, trimethoprim / sulfamethoxazole, batrafen, bifonazole, bleomycin, bortezomib, bryostatin-1, busulfan, calitiamycin, camptothecin, 10-hydroxycamptothecin, carbenicillin, caspofungin, Carmustine, cefaclor, cefazolin, cephalosporins, cefepime, ceftriaxone, cefotaxime, celecoxib, chlorambucil, chloramphenicol, ciprofloxacin, cisplatin, irinotecan (CPT-11), SN-38, carboplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dactinomycin, daunomyosingl clonide, daunorubicin, dexamethasone, diethylstilbestrol, diphtheria toxin, DNase I, doxorubicin, 2-pyrrolidoxorubicin (2P-DOX), doxycycline, cyanomorpholinodoxorubicin,Doxorubi single clonide, duocalmycin (DUBA), epirubic single clonide, ethinylestradiol, 7-ethyl-10-hydroxycamptothecin (SN-38), estramustine, estrogen receptor conjugate, etoposide, etoposide glucuronide, phosphate etoposide, erythrocycline, erythromycin, Flagyl, farnesyl-protein transferase inhibitor, floxuridine (FUdR), 3′,5′-O-dioleoyl-FudR (FUdR-dO), fludarabi Flutamide, Fluorouracil, Fluoxymesterone, Ganciclovir, Gentamicin, Geronin, Gemcitabine, Hydroxyprogesterone caproate, Hydroxyurea, Idarubicin, Ifosfamide, Imiquimod, Isoniazid, Itraconazole, Kanamycin, Ketoconazole, L-asparaginase, Leucovorin, Lomustine, Mechloretamine, Medroprogesterone acetate, Megestrol acetate, Melphalan, Mercaptopurine, 6-Mercaptopurine, Methotrexate, Mitox Santrone, Mithramycin, Mitomycin, Mitotane, Minocycline, Naftifine, Nalidixic acid, Neomycin, Navelbine, Nitrosourea, Nistatin, Lampirase, Oxacillin, Paromomycin, Penicillin, Pentamidine, Piperacillin / Tazobactam, Phenylbutyrate, Prednisone, Procarbazine, Paclitaxel, Pentostatin, Pokeweed antiviral protein, PSI-341, Seco-duocalmycin (SecoDUBA), Semustine, Rifabutin, Rifampin, Rimantadine, streptomycin, sulfamethoxazole, sulfasalazine, streptozocin, tamoxifen, taxanes, taxol, testosterone propionate, tetracycline, thalidomide, thioguanine, thiotepa, teniposide, topotecan, transplatinum, trimethoprim-sulfamethoxazole, uracil mustard, valacyclovir, vancomycin, vinblastine, vinorelbine, vincristine, zanamil, zithromycin, and (R)-5-chloro-N, 2 -[4-(4-methylpiperazin-1-yl)phenyl]-N 4Examples include, but are not limited to, -[(tetrahydrofuran-2-yl)methyl]pyrimidine-2,4-diamine((R)-9b).

[0088] In embodiments, D disclosed herein includes therapeutic agents for the treatment of cancer and non-cancer therapeutic agents. These therapeutic agents include small organic molecules. All hydroxyl group-containing therapeutic agents and amine-containing therapeutic agents for cancer treatment are included, for example, molecules that inhibit DNA replication (e.g., doxorubicin, epirubicin, calicheamicin, camptothecin), molecules that stabilize or disrupt microtubules (e.g., paclitaxel, docetaxel, epotilon), Na + / K + Examples of these therapies include molecules that affect the pump (e.g., strophantidine) and molecules that affect the function of the Golgi apparatus (e.g., nolisolide and its active derivatives). These therapeutic agents also include small inorganic molecules such as hydroxyl group-containing and amine-containing therapeutic agents used for cancer treatment, such as cisplatin and oxoplatin. Examples of linked antitumor agents include CO-doxorubicin and CO-strophantidine.

[0089] In other embodiments, D disclosed herein includes, but is not limited to, proteins. Examples include proteins of human and non-human origin, such as antibodies (e.g., trastuzumab), hormones (e.g., luteinizing hormone, follicle-stimulating hormone), cytokines (e.g., IL-6), growth factors (e.g., G-CSF), bacterial or plant toxins (e.g., Pseudomonas toxin, gelonin, ricin, abrin), and any type of tumor-targeting soluble protein; peptides, including artificial and natural peptides (e.g., ricin, TAT-related proteins that enhance cell penetration), which are toxic to tumor cells, alter the structure or function of tumor cells, or target other molecules to tumor cells or cells within a tumor that play a role in supporting tumor cells; nucleic acids such as RNA (e.g., antisense RNA, silencing RNA, toxin aptamers), DNA (e.g., naturally occurring or synthetic oligonucleotides), and high molecular weight structures (e.g., plasmids and viral vectors that express RNA or proteins toxic to tumor cells); particles of any size such as polymer-derived particles, protein-derived particles, metal-derived particles and inorganic particles (e.g., nanoparticles carrying a detectable label or imaging agent such as a fluorescent dye or radionuclide, or a therapeutic agent); and small molecules, including small inorganic and organic molecules that target cell surface receptors or otherwise bind to the surface of tumor cells or other accessible intracellular or extracellular components.

[0090] Therapeutic agents also include, for example, L-dopa, Ritalin, Cymbalta, Namenda and Gleevec. Agents that act on the central nervous system, such as L-dopa, Ritalin, Cymbalta, Namenda and Gleevec, are also included.

[0091] In other embodiments, D is an anticancer agent, an antineoplastic agent, or a cytotoxic molecule. In one embodiment, D is selected from the group consisting of amine group-containing antineoplastic agents such as monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and doxorubicin.

[0092] In one embodiment, D is selected from MMAE, MMAF, doxorubicin, cabazitaxel, docetaxel, paclitaxel, gemcitabine, imiquimod, SN-38, DUBA, seco-DUBA, (R)-9b, and gemcitabine.

[0093] In one embodiment, D is monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), exatecan, N-Me-L-Ala-maytansinol, gemcitabine, seco-duocarmycin (seco-DUBA), gemcitabine monophosphate, doxorubicin, cabazitaxel, docetaxel, paclitaxel, imiquimod, 7-ethyl-10-hydroxycamptothecin (SN-38), duocarmycin (DUBA), or (R)-5-chloro-N 2 -[4-(4-methylpiperazin-1-yl)phenyl]-N 4 -[(tetrahydrofuran-2-yl)methyl]pyrimidine-2,4-diamine ((R)-9b).

[0094] As mentioned above, in some embodiments, D is a diagnostic agent. A diagnostic agent is a molecule used in imaging tests such as magnetic resonance imaging (MRI), magnetic resonance tomography (MRT), positron emission tomography (PET), computed tomography (CT), single photon emission computed tomography (SPECT), optical imaging such as X-ray, and the like. A diagnostic agent is a detectable or traceable label. Examples of diagnostic agents for use in these tests include, but are not limited to, radioisotopes, dyes (including those using biotin-streptavidin complexes), enzymes, contrast agents, molecules such as fluorescent compounds or fluorochromes, paramagnetic ions (for MRI), and small molecules including, but not limited to, inorganic small molecules and organic small molecules that target cell surface receptors or otherwise bind to the surface of tumor cells or other accessible intracellular or extracellular components.

[0095] In certain embodiments, D is a radioisotope, imaging agent, fluorescent dye, near-infrared dye, enzyme, chemiluminescent agent, bioluminescent agent, paramagnetic ion, ultrasonic label, or radioacoustic label.

[0096] As mentioned above, D is -NR 2 It is bonded to L via a -, -S-, or -O- portion. For example, in some embodiments described herein, D is -NR 2 In another embodiment, D is bonded to L via -O-.

[0097] In a particular embodiment, the present disclosure provides an exemplary compound represented by the following formula. [ka] In a particular embodiment, D is MMAE.

[0098] In certain embodiments, the present disclosure provides an exemplary compound represented by the following formula: [ka]

[0099] In one embodiment, the present disclosure provides a pharmaceutical composition comprising the compounds of the present disclosure as described herein and pharmaceutically acceptable excipients, carriers, adjuvants, stabilizers, and / or diluents. The pharmaceutically acceptable excipients, carriers, adjuvants, stabilizers, diluents, etc., contained herein are determined by the composition to be administered and the method of administration of that composition. A wide variety of suitable formulations exist for pharmaceutical compositions comprising any pharmaceutically acceptable carriers, excipients, stabilizers, etc. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical field and are described, for example, in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Co., Easton, PA (1990).

[0100] For example, suitable pharmaceutical compositions for parenteral administration via intra-articular, intravenous, intramuscular, intradermal, intraperitoneal, and subcutaneous routes include aqueous and non-aqueous isotonic sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, and solutes to make the formulation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives. The compositions may be administered, for example, by intravenous infusion, orally, topically, intraperitoneally, intravesically, or subarachnoidally.

[0101] As used herein, the term “pharmaceutically acceptable salt” refers to a salt of a compound identified above that retains the desired biological activity and exhibits minimal or no unwanted toxicity. Examples of such salts include, but are not limited to, acid addition salts formed by inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.), as well as salts formed by organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid. Non-toxic pharmaceutically acceptable base addition salts include salts of bases such as sodium, potassium, calcium, and ammonium. In certain embodiments, the pharmaceutically acceptable salt is a sodium salt. In certain embodiments, the pharmaceutically acceptable salt is a potassium salt. Those skilled in the art will recognize a wide variety of non-toxic pharmaceutically acceptable addition salts.

[0102] The active compound (e.g., the compound of formula (I)) is contained in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective dose to the patient without causing serious toxicity to the patient being treated. For all of the above conditions, the dose of the active compound is in the range of about 0.01 to 300 mg / kg per day, preferably 0.1 to 100 mg / kg per day, and more generally in the range of 0.5 to about 25 mg per kilogram of body weight of the recipient per day. A typical topical dose is in the range of 0.01 to 3 wt / wt% in a suitable carrier. The effective dose range of a pharmaceutically acceptable derivative can be calculated based on the weight of the parent compound being delivered. If the derivative is active on its own, the effective dose can be estimated as described above using the weight of the derivative, or by other means known to those skilled in the art.

[0103] In certain embodiments, the dose of the active compound described herein is administered based on mg per kilogram of body weight of the recipient. In certain embodiments, the dose of the active compound described herein is administered once every 7 days (for example, once a week).

[0104] In certain embodiments, the dose of the active compound for all of the above-mentioned conditions may be in the range of about 0.1 to 20 mg / kg, or about 0.1 to 15 mg / kg, or about 0.1 to 10 mg / kg, or about 0.1 to 5 mg / kg, or about 0.5 to 20 mg / kg, or about 0.5 to 15 mg / kg, or about 0.5 to 10 mg / kg, or about 0.5 to 5 mg / kg, or about 1 to 20 mg / kg, or about 1 to 15 mg / kg, or about 1 to 10 mg / kg, or about 1 to 5 mg / kg, or about 2 to 20 mg / kg, or about 2 to 15 mg / kg, or about 2 to 10 mg / kg, or about 2 to 5 mg / kg. In certain embodiments described herein, the dose of the active compound is about 0.254 to about 0.45 mg / kg, about 0.6 to about 0.75 mg / kg, about 1.3 to about 1.6 mg / kg, about 3.4 to about 3.8 mg / kg, about 8 to about 10 mg / kg, or about 15 to about 20 mg / kg. In certain embodiments, the dose of the active compound of formula (I) is about 0.36 mg / kg, about 0.72 mg / kg, about 1.44 mg / kg, about about 9 mg / kg, or about 18 mg / kg. In certain embodiments described herein, the dose of the active compound is at least 0.36 mg / kg every 7 days, or at least 0.72 mg / kg every 7 days, or at least 1.44 mg / kg every 7 days, or at least 3.6 mg / kg every 7 days, or at least 9 mg / kg every 7 days, or at least 18 mg / kg every 7 days.

[0105] The pharmaceutical compositions disclosed herein may be manufactured by conventional mixing, dissolution, granulation, sugar coating, wet grinding, emulsification, encapsulation, encapsulation, or tableting processes. The pharmaceutical compositions may be prepared as liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquid before injection, or emulsions.

[0106] In certain specific aspects, the present disclosure also provides methods of using the disclosed compounds for therapeutic and diagnostic purposes. For example, the present disclosure provides a method for treating or alleviating a disease or condition, which can comprise administering an effective amount of one or more of the compounds described herein or one or more of the pharmaceutical compositions described herein to a subject in need thereof.

[0107] In an embodiment, the compound of the present disclosure delivers an anticancer agent to a selected tissue. The cancer can be lung cancer, breast cancer, colon cancer, ovarian cancer, prostate cancer, or melanoma.

[0108] Accordingly, in certain specific embodiments, the present disclosure provides a method of treating a patient with prostate cancer by administering to the patient an effective amount of a compound described herein or a pharmaceutical composition described herein. The amount of the compound and the regimen can be routinely determined using techniques accepted in the art.

[0109] In certain specific embodiments, the present disclosure provides a method for imaging one or more cancer cells (such as prostate cancer cells) in a patient by administering to the patient an effective amount of a compound described herein or a pharmaceutical composition described herein. The method may further comprise imaging the compound in vivo. The imaging can be performed using any imaging technique known in the art.

[0110] Another aspect of the present disclosure provides a compound having the structural formula (II)

Chemical Structure

[0111] In certain embodiments, each Y is independently selected from naturally occurring amino acids (e.g., L-amino acids) or unnaturally occurring amino acids (e.g., D-amino acids). In certain embodiments described herein, each Y is independently selected from naturally occurring amino acids (e.g., L-amino acids). For example, in certain embodiments, at least one Y is independently selected from alanine, glycine, isoleucine, leucine, proline, valine, phenylalanine, tryptophan, tyrosine, aspartic acid, glutamic acid, arginine, histidine, lysine, serine, threonine, cysteine, methionine, asparagine, or glutamine. In certain embodiments, at least one Y is selected from phenylalanine, tryptophan, tyrosine, or histidine. In certain embodiments, at least one Y is phenylalanine. In certain embodiments, at least one Y is tryptophan. In certain embodiments, at least one Y is 4-biphenylalanine. If Y is a naturally occurring amino acid, the side chain of such an amino acid may be further substituted. For example, the amino acid side chain may be -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl), halogen, C3-C 10 Cycloalkyl, C3-C8 heterocyclyl, aryl, heteroaryl, C3-C 10 It may be substituted with a cycloalkyl (C1-C6 alkyl), a C3-C8 heterocyclyl (C1-C6 alkyl), an aryl (C1-C6 alkyl), or a heteroaryl (C1-C6 alkyl), where R 10 and R 11 However, these are independently selected from hydrogen and C1-C6 alkyl groups.

[0112] In certain embodiments described herein, at least one Y has the following structure: [ka]

[0113] In a particular embodiment, each R 6 Each of these is independently (i) H, (ii) C1-C6 alkyl, (iii) hydroxy(C1-C6 alkyl), (iv) C1-C6 alkoxy(C1-C6 alkyl), (v) C2-C6 alkenyl, (vi) R 8 R 9 N(C1-C6 alkyl), (vii) halogen, (viii) halo(C1-C6 alkyl), (ix) carboxy(C1-C6 alkyl), (x) thio(C1-C6 alkyl), (xi) C1-C6 alkylthio(C1-C6 alkyl), (xii) R 8 R 9 NC(O)(C1-C6 alkyl), (xiii)C3-C 10 Cycloalkyl (C1-C6 alkyl), (xiv)C3-C8 heterocyclyl (C1-C6) alkyl, (xv) aryl (C1-C6 alkyl), (xvi) heteroaryl (C1-C6 alkyl), (xvii) R 17 C(O)NR 8 (C1-C6 alkyl), (xviii)C3-C 10 Selected from cycloalkyl, (xix)C3-C8 heterocyclyl, (xx)aryl, and (xvi) heteroaryl, where R 8 and R 9 R is independently selected from hydrogen and C1-C6 alkyl (e.g., C1-C4 alkyl or C1-C3 alkyl), and here R 17 However, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl), C3-C 10Cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), heteroaryl (C1-C6 alkyl), C3-C 10 The compounds are cycloalkyl, C3-C8 heterocyclyl, aryl, or heteroaryl. In certain embodiments, each cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, or -NR. 10 R 11 , R 10 R 11 N(C1-C6 alkyl), halogen, C3-C 10 Substituted with cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), or heteroaryl (C1-C6 alkyl), where R 10 and R 11 However, each is independently selected from hydrogen and C1-C6 alkyl (for example, C1-C4 alkyl or C1-C3 alkyl).

[0114] In certain embodiments described herein, at least one Y has the following structure: [ka]

[0115] In a particular embodiment, p is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, q is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, each R 7 These are, independently, H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, and -NR. 10 R 11 , R 10 R 11N(C1-C6alkyl), halogen, C3-C 10 Cycloalkyl, C3-C8 heterocyclyl, aryl, heteroaryl, C3-C 10 The compounds are selected from cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), or heteroaryl (C1-C6 alkyl), where each cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, or -NR. 10 R 11 , R 10 R 11 N(C1-C6alkyl), halogen, C3-C 10 Substituted with cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), or heteroaryl (C1-C6 alkyl), R 10 and R 11 Each of these is independently selected from hydrogen and a C1-C6 alkyl (e.g., a C1-C4 alkyl, or a C1-C3 alkyl). For example, in a particular embodiment, p is 1. In some embodiments described herein, q is 1. In a particular embodiment, p is 1, q is 1, and R 7 It is an aryl (for example, phenyl).

[0116] Each R 17 These are, independently, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, and -NR. 10 R 11 , R 10 R 11 N(C1-C6 alkyl), C3-C 10 Cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), and heteroaryl (C1-C6 alkyl), C3-C 10Selected from cycloalkyl, C3-C8 heterocyclyl, aryl, and heteroaryl, where R 10 and R 11 These are independently selected from hydrogen and C1-C6 alkyl (e.g., C1-C4 alkyl, or C1-C3 alkyl).

[0117] In certain embodiments described herein, at least one Y has the following structure: [ka]

[0118] In a particular embodiment, r is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, s is an integer between 1 and 7 (for example, 1 to 5, or 1 to 4, or 1 to 3). In a particular embodiment, each R 12 These are, independently, H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, and -NR. 10 R 11 , R 10 R 11 N(C1-C6alkyl), halogen, C3-C 10 Cycloalkyl, C3-C8 heterocyclyl, aryl, heteroaryl, C3-C 10 The compounds are selected from cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), or heteroaryl (C1-C6 alkyl), where each cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, or -NR. 10 R 11 , R 10 R 11 N(C1-C6alkyl), halogen, C3-C 10Substituted with cycloalkyl (C1-C6 alkyl), C3-C8 heterocyclyl (C1-C6 alkyl), aryl (C1-C6 alkyl), or heteroaryl (C1-C6 alkyl), R 10 and R 11 Each of these is independently selected from hydrogen and a C1-C6 alkyl (e.g., a C1-C4 alkyl, or a C1-C3 alkyl). For example, in a particular embodiment, r is 1. In some embodiments described herein, s is 1. In a particular embodiment, r is 1 and s is 1. In a particular embodiment, R 12 H is H.

[0119] In certain embodiments described herein, at least one X has the following structure: [ka]

[0120] In a particular embodiment, a is an integer between 1 and 4 (for example, 1 to 3). In a particular embodiment, b is an integer between 1 and 5 (for example, 1 to 4, or 1 to 3). In a particular embodiment, R 13 and R 14 Each of these is independently H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl) and halogens are selected, where R 10 and R 11 However, each is independently selected from hydrogen and C1-C6 alkyl (for example, C1-C4 alkyl, or C1-C3 alkyl). In a particular embodiment, R 13 and R 14 H is H.

[0121] In certain embodiments described herein, at least one Y has the following structure: [ka]

[0122] In a particular embodiment, c is an integer between 1 and 3 (for example, 1 or 2). In a particular embodiment, d is an integer between 1 and 4 (for example, an integer between 1 and 3). In a particular embodiment, R 15 and R 16 Each of these is independently H, -OH, -SH, -NO2, -COOH, C1-C6 alkyl, hydroxy(C1-C6 alkyl), C1-C6 alkoxy(C1-C6 alkyl), C2-C6 alkenyl, -NR 10 R 11 , R 10 R 11 N(C1-C6 alkyl) and halogens are selected, where R 10 and R 11 R is independently selected from hydrogen and C1-C6 alkyl (e.g., C1-C4 alkyl, or C1-C3 alkyl). In a particular embodiment, R 15 and R 16 H is H.

[0123] In certain embodiments described herein, at least one X has the following structure: [ka]

[0124] In certain embodiments, the compound of structural formula (II) is one of the earlier embodiments where n is an integer from 1 to 4 or from 1 to 3. In certain embodiments, n is 1 or 2. In other embodiments, n is 1. For example, in some embodiments, n is 1 and Y is phenylalanine. In other embodiments, n is 1 and Y has the structure shown below. [ka] In other embodiments described herein, n is 1 and Y has the following structure. [ka]

[0125] In certain embodiments, the compound of formula (II) is one of the previously described embodiments where m is an integer from 1 to 5. For example, in certain embodiments described herein, m is an integer from 1 to 4 or from 1 to 3. In certain embodiments, m is 1 or 2. In other embodiments of the present disclosure described herein, m is 1.

[0126] Another aspect of this disclosure relates to a compound having structural formula (III). [ka] or a pharmaceutically acceptable salt thereof, in the formula, Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R 2 It forms an amide with nitrogen having the following properties: m is an integer between 1 and 5. n is an integer from 1 to 4, and R 1 , R 2 , R 3 , and R 4 The present invention provides compound (I) or a pharmaceutically acceptable salt thereof, each independently being H, a C1-C6 alkyl group, or a protecting group.

[0127] The compound of structural formula (III) may have a specific stereochemistry. For example, in a particular embodiment, the present disclosure provides a compound having the following formula: [ka] Here, X, m, n, R1 , R 2 , R 3 , and R 4 However, this is as described with respect to equation (III).

[0128] In certain embodiments described herein, the compound of formula (III) includes X as defined in any of the earlier embodiments with respect to formula (I). In certain embodiments described herein, the compound of formula (III) includes n as defined in any of the earlier embodiments with respect to formula (I). In certain embodiments described herein, the compound of formula (III) includes m as defined in any of the earlier embodiments with respect to formula (I). In certain embodiments described herein, the compound of formula (III) includes R as defined in any of the earlier embodiments with respect to formula (I). 1 , R 2 , R 3 , and R 4 Includes.

[0129] Another aspect of this disclosure relates to a compound having structural formula (IV). [ka] or a pharmaceutically acceptable salt thereof, in the formula, D, -NR 2 A therapeutic or diagnostic agent bound to L via a -, -S-, or -O- moiety, L is an acid-cleavable linker, which combines to provide a carbamate moiety in formula (IV). Each Y is independently a natural or non-natural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety. m is an integer between 1 and 5, and We provide compounds where n is an integer from 1 to 4.

[0130] In certain embodiments described herein, the compound of formula (IV) includes D as defined in any of the earlier embodiments with respect to formula (I). In certain embodiments described herein, the compound of formula (IV) includes L as defined in any of the earlier embodiments with respect to formula (I). In certain embodiments described herein, the compound of formula (IV) includes Y as defined in any of the earlier embodiments with respect to formula (II). In certain embodiments described herein, the compound of formula (IV) includes m as defined in any of the earlier embodiments with respect to formula (II). In certain embodiments described herein, the compound of formula (IV) includes n as defined in any of the earlier embodiments with respect to formula (II). In certain embodiments described herein, the compound of formula (IV) includes R as defined in any of the earlier embodiments with respect to formula (I). 1 , R 2 , R 3 , and R 4 Includes.

[0131] Another aspect of this disclosure provides a method for synthesizing the compound of formula (I), as described above. This method synthesizes the compound of formula (V), as described herein. [ka] Alternatively, a pharmaceutically acceptable salt thereof may be used as the compound of formula (III). [ka] or a pharmaceutically acceptable salt thereof, wherein each X is independently a natural or non-natural n α-amino acid, and the C-terminus of X is R 2 It forms an amide with nitrogen, where m is an integer from 1 to 5, n is an integer from 1 to 4, and R 1 , R 2 , R 3 , and R 4 However, each of these is independently H, C1-C6 alkyl, or a protecting group.

[0132] Another aspect of this disclosure provides a method for synthesizing the compound of formula (IV) as described above. This method synthesizes the compound of formula (V) as described herein. [ka] Alternatively, a pharmaceutically acceptable salt thereof may be a compound of formula (II). [ka] or a pharmaceutically acceptable salt thereof, wherein each Y is independently a natural or non-natural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety, m is an integer from 1 to 5, and n is an integer from 1 to 4.

[0133] definition As used herein, the term “cell” means in vitro, ex vivo, or in vivo cells. In some embodiments, ex vivo cells may be a portion of a tissue sample excised from an organism such as a mammal. In some embodiments, in vitro cells may be cells in a cell culture. In some embodiments, in vivo cells are cells residing in an organism such as a mammal.

[0134] As used herein, the term “contact” refers to bringing together the indicated parts in an in vitro or in vivo system. For example, “contact” PSMA with a compound includes administering the compound described herein to a subject or patient, such as a human, and introducing the compound into a sample, for example, a cell preparation or purified preparation containing PSMA.

[0135] As used herein, the terms “subject” or “patient” are interchangeable and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, most preferably humans.

[0136] As used herein, the term “pharmaceutically acceptable salt” refers to both addition salts and solvates of pharmaceutically acceptable acids and bases. Such pharmaceutically acceptable salts include hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfinic acid, formic acid, toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, citric acid, tartaric acid, maleic acid, hydroiodic acid, alkanic acids such as acetic acid, and HOOC-(CH2) with n 0-4. n These include salts such as -COOH. Non-toxic pharmaceutically acceptable base addition salts include salts of bases such as sodium, potassium, calcium, and ammonium. In certain embodiments, the pharmaceutically acceptable salt is a sodium salt. Those skilled in the art will recognize a wide variety of non-toxic, pharmaceutically acceptable addition salts. [Examples]

[0137] The compounds described herein can be prepared using procedures well known to those skilled in the art and the procedures described herein. For example, the compound of structural formula (I) can be prepared using general procedures (below) and / or similar synthetic procedures. Those skilled in the art can adapt the reaction sequences and general procedures of Examples 1 and 2 to a desired target molecule. Of course, in certain circumstances, those skilled in the art will use different reagents to affect one or more of the individual steps or to use a particular protected version of one of the substituents. Furthermore, those skilled in the art will recognize that the compounds of this disclosure can be synthesized using entirely different routes.

[0138] General Information Regarding the compounds described in the following examples, 1 H and 311P NMR spectra were recorded using a Brucker Avance Neo 500 MHz spectrometer. High-resolution mass spectrometry (HRMS) spectra were obtained using an Applied Biosystems 4800 MALDI-TOF / TOF mass spectrometer. Scavenging resin, azido agarose, and DBCO agarose were purchased from Click Chemistry Tools. DBCO-NHS was purchased from AmBeed. PSMA-617 (bipivotide tetraxetan) was purchased from TargetMol (#T12573). All other chemical reagents used for synthesis and purification were purchased from AmBeed, Fisher, or Sigma Aldrich (unless otherwise noted) and used as supplied. Monomethyl auristatin E (MMAE, KP-20340) was purchased from eNovation Chemicals. Penicillin / streptomycin (P / S) was purchased from Corning. Fetal bovine serum albumin (FBS), HyClone® RPMI 1640 containing L-glutamine, and calcium and magnesium-free DPBS were purchased from Cytiva. Trypan blue was purchased from VWR. CELLSTAR® μCLEAR tissue culture treated, clear bottom, 96-well plates (#655098) were purchased from Greiner Bio-One. Trypsin 0.25%-EDTA 1mm (1X) and recombinant mouse PSMA / FOLH1 protein (#4946-Zn) were purchased from R&D Systems. Collagenase type I (0.25%) was purchased from STEMCELL Technologies. CellTiter-Glo® Luminescent Cell Viability Assay (G7572) reagent was purchased from Promega.

[0139] Example 1. Preparation and characterization of compound 1 (CTT2274) [ka]

[0140] Compound 3 (DBCO-Bip): Hp-phenyl-L-phenylalanine (272.92 mg, 1.13 mmol) was dissolved in 8 mL of anhydrous DMF in a 10 mL glass conical vial. DIPEA (656 μL, 486.6 µl) was added dropwise, followed by DBCO-NHS (303.25 mg, 753.6 µl) powder. The reaction mixture was stirred at 1100 Mot / min for 27 hours at room temperature. The reaction mixture was diluted with DCM (4 mL), transferred to a 50 mL recovery flask, and the solvent was removed by rotary evaporator at 60 °C. The crude substance was dissolved in Depositphotos (20 mL) and washed sequentially with 1N HCl (10 mL, 3 times), ddH₂O (10 mL, 2 times), and saline solution (20 mL, 2 times). The organic layer was dried over Na₂SO₄, volatile substances were removed under reduced pressure, and the resulting residue was dried under high vacuum. The crude substance was dissolved in DCM and purified by silica (20g) column chromatography (95:5 DCM:MeOH, containing 1% acetic acid) to obtain a solid in 87% yield.

[0141] Compound 6 (CTT1298): CTT1298 was synthesized as described in Ganguly, T. et al. A high-affinity[18F]-labeled phosphoramidate peptidomimetic PSMA-targeted inhibitor for PET imaging of prostate cancer. Nucl Med Biol 42, 780-787 (2015) and international patent application WO2014143736A1, each reference incorporated by reference.

[0142] Compound 4 (CTT2270): 1 equivalent of DBCO-Bip (78.78 mg, 149.04 μmol) was dissolved in anhydrous DMF (600 μL) and transferred to a 2 mL glass conical vial containing 5 molecular sieves. 1.5 equivalents of dried DIPEA (39 μL, 223.6 μmol) were added to the vial, followed by 1.3 equivalents of TSTU (58.3 mg, 193.8 μmol). The reaction mixture was stirred at room temperature at 1100 Mot / min for 12 minutes and monitored with silica TLC (90:10 EtoAc:ACN) to identify DBCO-BIP-NHS. 2 equivalents of CTT1298 (230.86 mg, 297.9 μmol) were dissolved in ddH2O (1.1 mL) and 2 equivalents of NaHCO3 (25 mg, 297.6 μmol) were added. The reaction mixture was frozen and freeze-dried overnight. The crude product was purified using Waters Sep-Pak with C18 (1 g, 6 cc, WAT036905) and C8 (1 g, 6 cc, WAT054570), and the products were eluted with 40% ACN / ddH2O and 30% ACN / ddH2O, respectively. The isolation yield was 18%.

[0143] Compound 5 (MMAE-PhosAm-azide) was synthesized as described in "Olatunji, FP et al. Modular Smart Molecules for PSMA-Targeted Chemotherapy". The following references are incorporated herein by reference: Mol Cancer Ther 21, 1701 (2022), Olatunji, FP, Herman, JW, Kesic, BN, Olabode, D. & Berkman, CEA click-ready pH-triggered phosphoramidate-based linker for controlled release of monomethyl auristatin E. Tetrahedron Lett 61, (2020), and Olatunji, FP, Herman, JW, Kesic, BN, Olabode, D. & Berkman, CECorrigendum to “A click-ready pH-triggered phosphoramidate-based linker for controlled release of monomethyl auristatin E” [Tetrahedron Lett. Volume 61, Issue 41, 8 October 2020, 152398]. Tetrahedron Lett 71, (2021). MMAE was purchased from eNovation Chemicals (Cat #KP-20340).

[0144] Compound 1 (CTT2274): In a 1.5 mL polypropylene microcentrifuge tube, 1.1 equivalents of MMAE-PhosAm-azide (28.83 mg, 20.6 μmol) were dissolved in ddH2O (60 μL) containing 1.5 equivalents of NaHCO3 (27.4 μmol), and the mixture was centrifuged with pulses at 14.5 rcf for 10 seconds. In another 1.5 mL microcentrifuge tube, 1 equivalent of CTT2270 (22.11 mg, 18.3 μmol) was dissolved in ddH2O (160 μL) containing 1.5 equivalents of NaHCO3 (27.4 μmol). CTT2270 was added dropwise to the MMAE-PhosAm-azide mixture, and the mixture was stirred at room temperature at 500 Mot / min for 1 hour. Unreacted DBCO-Bip and MMAE-PhosAm-azide were removed using azido agarose and DBCO agarose (Click Chemistry Tools), respectively, as previously described for CTT227X. The final removed material was desalted with C8 Waters Sep-Pak (1 g, 6 cc, WAT054570) prepared with ACN, equilibrated with ddH2O, and eluted with 35% ACN. After evaporation of ACN, the pooled fraction was frozen, and the product was freeze-dried overnight. The isolation yield was 71%. 1 HNMR and 31 The PNMR results are shown in Figures 1A and 1B.

[0145] Example 2. Preparation and characterization of compound 2 (CTT227X) [ka]

[0146] Compound 2 (CTT227X): 1.1 equivalents of DBCO-Bip (16.86 mg, 12 umol) were dissolved in MeOH (120 μL) in a 1.5 mL polypropylene microcentrifuge tube. In another 1.5 mL microcentrifuge tube, 1 equivalent of MMAE-PhosAm-azide (15.11 mg, 10.8 μmol) was dissolved in ddH2O (120 μL) containing 3.5 equivalents of NaHCO3 (38.3 μmol), and the mixture was centrifugated with pulses at 14.5 rcf for 10 seconds. MMAE-PhosAm-azide was added dropwise to DBCO-Bip and stirred at room temperature at 400 Mot / min for 4 hours and 5 minutes. A cloudy precipitate began to form immediately after the addition of MMAE-PhosAm-azide to DBCO-Bip, so an additional 300 μL of MeOH was added to the reaction to aid in the dissolution of the material. The final MeOH content in the reaction product was 78%. Unreacted DBCO-Bip and MMAE-PhosAm-azide were removed using azido agarose and DBCO agarose (Click Chemistry Tools), respectively. Briefly, 1 mL of the removal resin (i.e., 2 mL of 50% agarose slurry) was washed twice with 1 mL of a solution of 10% DMSO, 30% ethanol, and 100 mM NaHCO3. The entire reaction mixture was added to the rinsed resin and rotated on a rotisserie at room temperature for 35 minutes. The resin filtrate was collected, and the resin was washed with ddH2O (1 mL, 3 times), with each washing solution collected along with the original filtrate. The samples were frozen and freeze-dried overnight. The final swept resin, conditioned with ACN, was desalted with C18 Waters Sep-Pak (1g, 6cc, WAT036905), equilibrated with ddH2O, and eluted with 40% ACN. After evaporating off the ACN, the pooled fraction was frozen, and the product was freeze-dried overnight. The isolation yield was 58%.

[0147] Example 3. Preparation and characterization of compound 8 (CTT2101) [ka]

[0148] Compound 7 (CTT1400): CTT1400 was synthesized as described in "Olatunji, FP et al. Modular Smart Molecules for PSMA-Targeted Chemotherapy". Mol Cancer Ther 21,1701(2022),Olatunji,FP,Herman,JW,Kesic,BN,Olabode,D.&Berkman,CEA click-ready pH-triggered phosphoramidate-based linker for controlled release of monomethyl auristatin E.Tetrahedron Lett 61, (2020.), and Olatunji,FP,Herman,JW,Kesic,BN,Olabode,D.&Berkman,CECorrigendum to “A click-ready pH-triggered phosphoramidate-based linker for controlled release of monomethyl auristatin E”[Tetrahedron Lett.Volume 61,Issue 41,8 October 2020,152398].Tetrahedron Lett 71,(2021). MMAE was purchased from eNovation Chemicals (Cat #KP-20340). References are incorporated herein by reference. Therefore, CTT1298 was dissolved in ddH2O to prepare a 0.43 M solution. 125 μL of this solution was added to a 1 mL conical vial. 1 M TEA-Bicarb buffer was added to the 1 mL conical vial containing the CTT1298 solution. 1.8 equivalents of DBCO-PEG4-NHS were dissolved in DMSO (to prepare a 0.26 M solution) and added dropwise to the vial. The reaction mixture was vigorously stirred overnight at 4°C. The reaction mixture was then purified by preparative HPLC and dried by lyophilization. Before lyophilization, 1.2 equivalents of NaHCO3 were added to neutralize the pH.

[0149] Compound 8 (CTT2101): In a 1.5 mL polypropylene microcentrifuge tube, 1.1 equivalents of MMAE-PhosAm-azide (21.51 mg, 15.4 umol) were dissolved in ddH2O (60 μL) containing 1.5 equivalents of KHCO3 (20.85 umol), and the mixture was centrifugated with pulses at 14.5 rcf for 10 seconds. In another 1.5 mL microcentrifuge tube, 1 equivalent of CTT1400 (18.21 mg, 13.9 umol) was dissolved in ddH2O (110 μL) containing 1.5 equivalents of KHCO3 (20.85 umol). CTT1400 was added dropwise to the MMAE-PhosAm-azide mixture, and the mixture was stirred at room temperature at 500 Mot / min for 1 hour. Unreacted DBCO-Bip and MMAE-PhosAm-azide were removed using azido agarose and DBCO agarose (Click Chemistry Tools), respectively, as described previously for CTT227X, except that 100 mM KHCO3 was used instead of 100 mM NaHCO3 for washing the swept resin. The mixture was conditioned with ACN, equilibrated with ddH2O, and desalted with C8 Waters Sep-Pak (1 g, 6 cc, WAT054570) eluted with 20% ACN. After evaporating off the ACN, the pooled fraction was frozen, and the product was freeze-dried overnight. The isolation yield was 71%. This reaction is shown in Scheme 5.

[0150] Example 4. IC 50 Determination and analysis of inhibition mechanisms IC using human PSMA 50 The determination of the inhibitory factors and the analysis of the inhibition modes are routinely performed as described above [4-6]. In the case of recombinant mouse PSMA, the final reaction conditions were based on monitoring product formation over time at 37°C by HPLC analysis, creating a time curve, and identifying the linear range. 50In the study, 0.5 μL of recombinant mouse PSMA (0.22 μg) was diluted 1:1999 with 50 mM Tris pH 7.4 containing 1% Triton X-100. The reaction was prepared with 175 μL of 50 mM Tris pH 7.4, 25 μL of 10 μM substrate (N-[4-(phenylazo)-benzoyl]-glutamyl-g-glutamic acid, PABGgG), and 25 μL of inhibitor or Tris buffer. The reaction was initiated with 25 μL of recombinant mouse PSMA diluted 1:1999 (total reaction volume 250 μL) and incubated at 37°C for 15 minutes. The final inhibitor concentration range was 0.1 nM to 10 μM. In the analysis of the inhibition mode, the mouse PSMA concentration was 100 times higher than in the IC50 test, and the determined IC50 was higher. 50 The inhibitor was pre-incubated at 37°C for 10 minutes with an inhibitor concentration 10 times higher than that or with 50 mM Tris buffer pH 7.4 (total sample volume 40 μL). After pre-incubation, the enzyme inhibitor / buffer sample was rapidly diluted with 1 mM substrate in 50 mM Tris pH 7.4 containing 1% Triton buffer (3960 μL). 200 μL of the sample was taken every 5 minutes for 1 hour, and product formation was monitored. All reactions were quenched with 25 μL of 2.5% trifluoroacetic acid in methanol, and samples for HPLC analysis were prepared as described previously [6] [6]. IC of the compound 50は We compared both human PSMA (hPSMA) and mouse PSMA (mPSMA). For hPSMA, we used the IC of CTT2101. 50 The value is 3.97±0.126nM, CTT2274 IC 50 The value was 3.97 ± 0.134 nM. For mPSMA, the IC of CTT2274 was used. 50 The value was 105 ± 4.79 nM. MMAE is less than 10 μM compared to hPSMA. 50 The values ​​were shown. CTT227X and MMAE did not show significant inhibition against mPSMA at 10 μM. The coupling of CTT2101 and CTT2274 was determined to be irreversible. Figures 2A and 2B show the IC of CTT2274 against hPSMA and mPSMA, respectively. 50 The curve is shown, and Figure 2C shows the IC of CTT2101 relative to hPSMA. 50 It shows a curve.

[0151] Example 5. Western blot analysis PSMA protein expression levels in the JAX Laboratories PDX prostate adenocarcinoma model (TM00298) and in the PC3, C4-2B, and PC3-PIP prostate cancer cell lines were examined by Western blot analysis. C4-2B cells were kindly provided by Eva Corey of the University of Washington. PC3, C4-2B, and PC3-PIP cells were cultured at 37°C under 5.0% CO2 conditions until 70-80% confluence was reached, in medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (P / S) in RPMI1640 containing L-glutamine and DPBS (HyClone®) without calcium and magnesium.

[0152] Frozen tumor samples of approximately 6 mm square, collected from a PDX prostate adenocarcinoma model, were thawed in 1 mL of RPMI-1640 medium (1% P / S, 2% FBS) at 37°C for 5 minutes. The tissue in the medium was centrifuged at 65 rcf for 3 minutes, and the medium was replaced with 2 mL of collagenase type 1 (0.25%, STEMCELL Technologies), and incubated at 37°C for 1 hour. Cell disruption was performed by passing the cells three times through a 20-gauge, 1-inch needle attached to a 3 cc syringe. Collagenase type 1 was neutralized with 1 mL of 5% FBS RPMI-1640 (1% P / S). The digested cell samples were centrifuged at 65 rcf for 5 minutes, and the supernatant was discarded. The digested cells were resuspended in PBS using a pipette, centrifuged at 65 rcf for 3 minutes, and the supernatant was discarded. Cells were lysed in 1 mL of ice-cold lysis buffer (150 mM NaCl, 1% NP-40, 50 mM Tris-HCl, pH 7.4) supplemented with a 1× APExBIO protease inhibitor cocktail (#K1019) in DMSO containing EDTA, and incubated at 4°C for 20 minutes. The lysed cells were centrifuged at 4°C and 14500 rcf for 10 minutes, and the supernatant was collected for protein quantification.

[0153] PC3, C4-2B, and PC3-PIP cells were cultured in complete medium (RPMI-1640, 10% FBS, 1% penicillin / streptomycin) in T-75 flasks at 37°C and 5.0% CO2 until 70–80% confluence was reached. Cells were washed with cold (4°C) PBS and incubated in 0.5 mL of ice-cold lysis buffer (150 mM NaCl, 1% NP-40, 50 mM tris-HCl, pH 7.4) supplemented with a 1X APExBIO protease inhibitor cocktail in DMSO containing EDTA. Cells were gently scraped from the flask and transferred to a microcentrifuge tube and incubated at 4°C for 20 minutes. Cells were centrifuged at 4°C and 14,500 rcf for 10 minutes. The supernatant was collected and proteins were quantified by Bio-Rad protein assay. Protein standard curves were prepared using bovine serum albumin according to the manufacturer's standard procedure for microtiter plates. Absorbance at 595 nm was measured using a Fluostar Omega microplate reader with Omega software version 1.02 and Mars Data Analysis Software Program version 1.10 (BMG Labtech).

[0154] Cell lysates were prepared in sample buffer (50 mM Tris-HCl pH 6.5, 2% SDS, 8% glycerol, 0.1% bromophenol blue, 5.3% β-mercaptoethanol) and boiled at 100°C for 3 minutes. Proteins were separated on a 10% polyacrylamide gel in Tris-glycine SDS buffer (25 mM Tris pH 8.3, 192 mM glycine, 0.1% SDS) by SDS-PAGE at 200 V for 35 minutes. Proteins were transferred to a 0.2 μm nitrocellulose membrane in Towbin buffer (25 mM Tris, 192 mM glycine, 20% methanol pH 8.3) at 100 V for 1 hour. The membranes were washed with Tris-buffered saline (TBS) (20 mM Tris pH 7.6, 150 mM NaCl) and blocked at room temperature for 30 minutes with 3% gelatin in TBS. The membranes were washed with Tris-buffered saline-Tween 20 (TBS-T) (20 mM Tris-HCl pH 7.5, 500 mM NaCl, 0.05% Tween 20). The membranes were probed using 1:1000 dilutions of PSMA(D4S1F) rabbit mAb (#12702, Cell Signaling Technology) and 1:1000 dilutions of GAPDH(D16H11)XP rabbit mAb (#5174, Cell Signaling Technology), respectively, diluted in TBS-T containing 1% gelatin. Proteins were detected colorimetrically using the BioRad Immun-Blot Goat Anti-Rabbit IgG(H+L)-AP Assay Kit (#1706460) according to the manufacturer's instructions.

[0155] PSMA expression in PC3, PC3-PIP, C4-2B cell lines, and PDX tumors was measured by Western blotting. The Western blots are shown in Figure 3. The PC3 cell line did not express PSMA, but the PC3-PIP, C4-2B cell lines, and PDX tumor samples did express PSMA.

[0156] Example 6. In vitro cytotoxicity The cell viability-reducing effect of each CTT compound was measured in prostate cancer cell lines with varying PSMA expression levels using the Cell-Titer-Glo Luminescent Cell Viability Assay (Promega). PC3 was PSMA-negative, C4-2B was PSMA-positive, and PC3-PIP was PSMA-positive, as confirmed by Western blot analysis. All cell lines were cultured in complete medium (RPMI-1640, 10% FBS, 1% penicillin / streptomycin). Cells were detached with trypsin / EDTA, counted with trypan blue, and serially diluted with complete medium to prepare working concentrations between 100,000 cells / mL and 1,560 cells / mL. 100 μL of cells were seeded in triple layers in each drug or control well on a clear, flat-bottomed 96-well tissue culture plate (Grenier Bio-One #). One 96-well plate was used per cell line. Each cell viability assay was repeated three times (n=3) using different cell passages. The final cell count in each experimental well was 2500 cells / well. Cell standard curves were created for each cell line in the range of 156–10000 cells / well to verify the linear relationship of chemiluminescence signals with increasing cell count. Cells were bonded overnight at 37°C and 5.0% CO2 before drug treatment. MMAE was dissolved in DMSO, and all CTT compounds were dissolved in 100 mM HEPES pH 7.4. 100 μL of either complete medium containing 0.2 nM–20 nM of drug, vehicle (0.2% DMSO in medium or 2% 100 mM HEPES pH 7.4 in medium), or complete medium was added to each well to a total volume of 200 μL. Empty wells were filled with 200 μL of phosphate-buffered saline (PBS). The final drug concentrations were 0.1 nM, 1 nM, and 10 nM for MMAE, CTT2101, CTT2274, and CTT227X, respectively. The final proportions of DMSO and 100 mM HEPES pH 7.4 buffer in the culture medium were 0.1% and 2%, respectively, in the complete medium, and these were maintained throughout the assay. Only seeded cells were treated in complete medium alone to generate a cell standard curve. During the 72-hour drug treatment period, 150 μL was removed from each well every 24 hours and replaced with fresh drug, vehicle, or complete medium.Given that the pH of the culture medium may begin to decrease as cells grow, changing the medium every 24 hours was expected to reduce the possibility of premature drug release during the assay. After a 72-hour drug treatment period, 100 μL was removed from all wells, and the 96 plate was equilibrated at room temperature (RT) for 30 minutes. 100 μL of CellTiter-Glo reagent was added to each well, and chemiluminescence was measured using a Fluostar Omega microplate reader with Omega software version 1.02 and Mars Data Analysis Software Program version 1.10 (BMG Labtech) according to the manufacturer's instructions. Viability (%) relative to control was calculated as follows:

number

[0157] AVG LUM is the calculated average value of the measured luminescence.

[0158] The in vitro cytotoxicity of CTT2101, CTT2274, CTT227X, and MMAE was tested against three prostate cancer cell lines: PC3, PC3-PIP, and C4-2B. In the PC3 cell line, MMAE was the only compound to show a dose-responsive effect; CTT2101, CTT2274, and CTT227X showed nonspecific killing (Figure 4A). In the PC3-PIP cell line, CTT2101, CTT2274, and MMAE showed dose-responsive effects. At a drug concentration of 1 nM, MMAE was significantly more lethal than any of the other drugs, resulting in a cell viability of 30.33%. At a drug concentration of 10 nM, MMAE showed the lowest cell viability (25.33%), while CTT227X showed no measurable effect, resulting in a cell viability of 89.67%. CTT2274 showed the second lowest cell viability (47.33%), while treatment with CTT2101 resulted in a cell viability of 63.67% (p=0.03) (Figure 4B). Finally, in the C4-2B cell line, CTT2101 and CTT2274 did not show any evaluable effect until the drug concentration reached 10 nM, whereas MMAE nearly completely killed cells at drug concentrations of 1 nM and 10 nM (Figure 4C). At 10 nM, CTT2101, CTT2274, and CTT227X showed cell viability of 78%, 38.67%, and 91.33%, respectively. CTT2274 caused significantly more cell death than CTT2101 at this concentration (p=0.003).

[0159] Example 7. Comparative evaluation of the effectiveness of CTT2101 with MMAE. This study was conducted at The Jackson Laboratory in Sacramento, California. Male NSG mice (stock #005557) aged 6-8 weeks were subcutaneously inoculated into the right flank with 40 μL of xenografted prostate cancer tumor from patient TM00298. 29Based on tumor volume, mice were randomly assigned to one of three treatment groups: CTT2101, MMAE, or PBS. A t-test was performed to confirm that there were no statistically significant differences between the groups. Thirty animals (n=10 / group) were administered either 0.8 mg / kg of CTT2101 in 100 μL of 1×PBS (equivalent to 0.2 mg / kg of MMAE), 0.2 mg / kg of MMAE in 100 μL of 1×PBS, or 100 μL of 1×PBS via tail vein injection. The 0.2 mg / kg dose of MMAE was determined based on previously published studies and the known LD50 of MMAE when administered intravenously to mice. 50 The dosage was selected based on 1-2 mg / kg. Animals received a total of 6 doses, once a week starting from day 0. From day 39, animals received a total of 6 doses every 3 days. During the study period, animals were observed daily for signs of illness and death. Body weight was measured twice a week. Tumor volume (width squared × 0.5 × length) was measured twice a week using calipers. Animals were given saline and Nutra Gel as supportive therapy as needed, according to the instructions of the veterinary staff. Tumor volume was 2,000 mm 3 Animals were euthanized if the body condition score exceeded a certain threshold, if the body condition score was 2 or lower, or if ulceration or necrosis of the tumor was confirmed. This study was conducted blindly.

[0160] The efficacy of CTT2101 was evaluated compared to MMAE using a mouse prostate cancer PDX model. All tumor volumes at baseline ranged from 49.87 to 308.42 mm². 3 The mean tumor volume before the first dose was 175.11 ± 69.24 mm² in the CTT2101 group, MMAE group, and PBS group, respectively. 3 , 175.94±77.01mm 3 , 175.94±73.78mm 3The animals maintained a stable body weight throughout the study period (Figure 5A). Looking at the changes in tumor growth over time, it is clear that MMAE provides more significant and sustained tumor suppression over time compared to CTT2101, even when the administration frequency is increased to once every three days (Figure 5B). Figures 5C-5E show spaghetti plots of tumor growth for CTT2101, MMAE, and PBS, respectively. Based on this data, it is possible to determine the time it takes for the tumor load at the start of treatment to reach the exponential growth phase (Figure 5C).

[0161] The median survival time was 56 days, 70 days, and 48 days for the CTT2101 group, MMAE group, and PBS group, respectively. There was no statistically significant difference in survival time between CTT2101 and PBS. Statistically significant differences in survival time were observed when comparing the CTT2101 group with the MMAE group (p=0.0017) and the MMAE group with the PBS group (p<0.0001) (Figure 5F). However, when the CTT2101 group mice were analyzed based on tumor volume at the start of treatment, the tumor was 200 mm 3 The median survival time for mice with a tumor size of less than 200 mm was 63 days, while for mice with a tumor size of 200 mm... 3 The median survival time for mice exceeding this size was only 48 days, and this difference is statistically significant (p=0.0052) (Figure 5G). In the MMAE group, there was no statistically significant difference in survival time based on initial tumor volume (data not shown). Furthermore, the tumor size at the start of administration was 200 mm 3 Mice in the CTT2101 group, which had tumors smaller than 50%, showed delayed tumor regrowth after treatment. In mice with small initial tumors, no significant increase in tumor volume was observed until around day 35, whereas in animals with larger tumors at the start of administration, an increase in tumor volume was observed around day 21. The same trend was observed in the MMAE group, with animals with small tumors showing tumor regrowth from around day 42, and animals with large tumors showing tumor regrowth from around day 28 (Figures 6A-6D).

[0162] Example 8. Comparative evaluation of the effectiveness of CT2274 with CTT2101 and MMAE. This study was conducted at The Jackson Laboratory in Sacramento, California. Male NSG mice (stock #005557) aged 6-8 weeks were subcutaneously inoculated into the right flank with 40 μL of xenografted prostate cancer tumor from patient TM00298. Based on tumor volume, mice were randomly assigned to one of the treatment groups: CTT2101, CTT2274, MMAE, or PBS. A t-test was performed to confirm that there were no statistically significant differences between the groups. Forty animals (n=10 / group) were administered via tail vein injection: 3.8 mg / kg of CTT2101 (equivalent to 1 mg / kg of MMAE) in 100 μL of 1X PBS, 3.6 mg / kg of CTT2274 (equivalent to 1 mg / kg of MMAE) in 100 μL of 1X PBS, 1 mg / kg of MMAE in 100 μL of 1X PBS, or 100 μL of 1X PBS. The animals received a total of six doses, once a week, starting from day 0. During the study period, the animals were observed daily for signs of illness and death. Body weight was measured twice a week. Tumor volume (width squared × 0.5 × length) was measured twice a week using calipers. From day 86, body weight and tumor volume measurements were switched to once a week. The animals received saline and Nutra Gel as supportive therapy as needed, following the instructions of the veterinary staff. Tumor volume reached 2,000 mm². 3 Animals were euthanized if the body condition score exceeded a certain threshold, if the body condition score was 2 or lower, or if ulceration or necrosis of the tumor was confirmed. This study was conducted blindly.

[0163] The efficacy of CTT2274 over CTT2101 and MMAE was evaluated using the same patient-derived xenograft prostate cancer model used in the initial efficacy trial. In this trial, the dose of MMAE was increased fivefold to 1 mg / kg, and CTT2101 and CTT2274 were also increased accordingly. Throughout the trial, mice administered with CTT2101, CTT2274, and PBS showed no change in body weight, while mice administered with MMAE showed a sawtooth pattern in body weight. A transient decrease in body weight was consistently observed on day 2 after administration. After discontinuation of treatment on day 35, the body weight of mice in the MMAE group steadily decreased and then increased (Figure 7A).

[0164] All tumor volumes at the start of the study ranged from 62 to 172 mm². 3 The mean tumor volume before the first dose was 98.90 ± 33.41 mm² in the CTT2101, CTT2274, MMAE, and PBS groups, respectively. 3 98.95±29.25mm 3 98.60±32.05mm 3 98.83±32.99mm 3 As observed in the initial trial, CTT2101 showed mild tumor suppression compared to PBS, but there was no statistically significant difference between the two groups. In contrast, CTT2274 and MMAE showed statistically significant tumor suppression compared to PBS, and importantly, there was no statistically significant difference between CTT2274 and MMAE throughout the trial (Figure 7B). Figures 7C–7F show tumor growth spaghetti plots for the CTT2101, CTT2274, MMAE, and PBS groups, respectively.

[0165] The mean survival time was 53 days in the CTT2101 group and 48 days in the PBS group. In the CTT2274 group, there were no mice that met the euthanasia criteria or died due to tumor burden. In the MMAE group, two mice died (due to drug toxicity) and one died due to tumor burden. There was no statistically significant difference in survival time between the CTT2101 group and the PBS group (p=0.09). However, mice treated with CTT2274 survived significantly longer than those treated with CTT2101 (p<0.0001) and PBS (p<0.0001) (Figure 7G). From this study, the following conclusions were drawn. 1) CTT2274 3.6 mg / kg showed a long-term tumor suppression effect comparable to MMAE even after discontinuation of treatment. 2) No obvious signs of toxicity (changes in body weight or other physical symptoms) appeared with CTT2274 3.6 mg / kg. 3) CTT2101 was less effective than CTT2274 in both tumor suppression and overall survival.

[0166] Example 9. Safety / toxicity of CTT2274 compared to MMAE This study was conducted at The Jackson Laboratory in Sacramento, California. Male C56BL / 6J mice (stock number 000664) aged 6–8 weeks were used (n=3 / group / time point). Two weeks prior to administration, 200 μL of blood was collected from the retroorbital venous plexus of the mice. 150 μL of blood was treated with serum, and 50 μL of whole blood was collected in a tube containing K2EDTA. Two days prior to administration, the mice were divided into groups according to body weight. On day 0, mice were administered a single dose via tail vein injection of 100 μL of 1X PBS containing either 3.6 mg / kg of CTT2274 (equivalent to 1 mg / kg of MMAE), 36 mg / kg of CTT2274 (equivalent to 10 mg / kg of MMAE), 1 mg / kg of MMAE, or 10 mg / kg of MMAE. Blood samples were collected posteriorly orbitally at 8, 24, and 168 hours after administration and processed as described above. Terminal blood sampling was performed by cardiac puncture at 336 hours after administration. All samples were sent to the Comparative Pathology Laboratory at the University of California, Davis for analysis. Complete blood counts and comprehensive chemistry tests were performed on all samples. Body weight and clinical observations were recorded daily. This study was conducted blindly. Figure 8 shows the major blood chemistry and hematological findings, respectively. The black dotted lines indicate the reference range with a 95% confidence interval. It is well understood that reference ranges vary depending on sex, strain, age, and blood collection route. Therefore, this data and reference range should be used as a guide for future study designs. Both MMAE and CTT2274 were found to be toxic and lethal at high doses. Mice in the 3.6 mg / kg CTT2274 group maintained a stable body weight throughout the study period, while all other groups showed weight loss. The 1 mg / kg MMAE group showed signs of recovery by day 14 of the study. In either the 1 mg / kg MMAE or 3.6 mg / kg CTT2274 groups, no mice were confirmed dead or required euthanasia during the study.

[0167] Several significant changes were observed in CBC and blood chemistry parameters. Hyperglycemia was observed in both the 1 mg / kg MMAE and 3.6 mg / kg CTT2274 groups. Although the exact mechanism is unknown, hyperglycemia has also been observed in patients treated with Padcev® and Adcetris®, suggesting that glucose should be carefully monitored in future trials. Potassium and phosphorus levels increased by day 2 in both the 1 mg / kg MMAE and 3.6 mg / kg CTT2274 groups, but interestingly, these values ​​did not return to baseline by the end of the trial. While elevated potassium and phosphorus levels may be due to acute kidney injury, serum creatinine and BUN levels were normal, indicating a low probability of AKI. Insulin resistance and hemolysis can also cause elevated potassium and phosphorus levels. Importantly, the 1 mg / kg MMAE group showed variability, while the 3.6 mg / kg CTT2274 group showed only minor variability. Padcev®, Adcetris®, and Polivy® all list thrombocytopenia as a potential side effect. Moderate lymphopenia was observed in both the 1 mg / kg MMAE group and the 3.6 mg / kg CTT2274 group, but it was more pronounced in animals treated with MMAE. Lymphopenia was also observed in patients treated with Padcev®, Tivdak®, and Polivy®, further indicating myelosuppression. In addition, a decrease in red blood cell count was observed on day 7 in the 1 mg / kg MMAE group, suggesting possible myelosuppression, while the values ​​in the 3.6 mg / kg CTT2274 group did not change (the changes in hemoglobin and hematocrit were similar to the changes in red blood cells). Alkaline phosphatase (ALP) levels were significantly elevated 8 hours after administration of 1 mg / kg MMAE in mice, but returned to a similar level to the 3.6 mg / kg CTT2274 group after 24 hours. Finally, while the absolute number of neutrophils changed dramatically in the 1 mg / kg MMAE group, no clinically significant changes were observed throughout the study in the 3.6 mg / kg CTT2274 group. No significant changes were observed in any other CBC or chemical parameters.

[0168] While this safety / toxicity study has limitations, several important pieces of information were obtained: 1) 36 mg / kg CTT2274 and 10 mg / kg MMAE were acutely toxic and caused death in animals. 2) 3.6 mg / kg CTT2274 showed minimal safety issues compared to the equivalent dose of 1 mg / kg MMAE. 3) The hematological and clinical chemistry variability observed at the 3.6 mg / kg dose of CTT2274 is consistent with previously published data comparing it to MMAE ADCs.

[0169] In summary, CTT2274 3.6 mg / kg was shown to be effective and possess a favorable safety profile for continued drug development. Table 1 outlines how CTT2274 compares to free MMAEs, competing ADCs, and similar MMAE SMDCs based on key drug commercialization aspects and desirability in future compound development. [Table 1]

[0170] Consideration MMAEs are a promising treatment option for PCa patients, but safely and effectively delivering them remains a challenge. ADCs utilizing MMAEs rely on unstable linkers, cause significant adverse events, and have high manufacturing costs. Here, the efficacy and safety of CTT2274 are demonstrated.

[0171] In in vitro cytotoxicity studies, as expected, only MMAE induced cell death in the PSMA-PC3 cell line. CTT2101 and CTT2274 were effective in both PSMA+ cell lines at a concentration of 10 nM, but CTT2274 resulted in lower cell viability. Importantly, this indicates that selective uptake is observed, as seen in CTT227X, even after removal of the PSMA binding moiety, no cell death occurs.

[0172] In the initial efficacy trial, CTT2101 was evaluated against MMAE using a PDX model of prostate cancer. CTT2101 showed moderate tumor suppression at a dose of 0.8 mg / kg (equivalent to 0.2 mg / kg of MMAE). Animals were initially administered once a week for 6 weeks. At the end of that cycle, the administration frequency was increased to once every 3 days for a total of 6 times, and it was evaluated whether further suppression of tumor growth was achieved by the administration. At this point, the mean tumor volume was 943.74 ± 462.06 mm in the CTT2101 group, MMAE group, and PBS group, respectively. 3 , 416.92±260.60mm 3 , 1175.68±570.90mm 3 It was found that increasing the administration frequency to once every three days was ineffective for such large tumors. It became clear that tumor volume at the start of treatment significantly impacts treatment efficacy and overall survival. With clinical applicability in mind, CTT2101 was modified to include a biphenyl motif, and it was expected that this would extend the circulating time to maintain a once-weekly administration regimen.

[0173] In the second efficacy trial, CTT2101, CTT2274, and MMAE were compared in the same PDX model of prostate cancer. Based on the moderate therapeutic effect of CTT2101 observed in the first efficacy trial, the dose was increased to the equivalent of 1 mg / kg MMAE. Furthermore, according to information obtained from the first trial, the tumor volume at the start of treatment was 200 mm². 3 It did not exceed [value missing]. This combination of modifications (biphenyl motif, increased dose, control of tumor volume at the start of treatment) proved to be very effective. MMAE caused a sawtooth-like change in body weight, which decreased consistently after 2 days of administration and eventually steadily. On the other hand, mice treated with CTT2274 did not show any significant change in body weight throughout the study period. Furthermore, the first mouse to die in the MMAE group had a tumor of 218.02 mm. 3Since there was no other explanation, it was considered highly likely that the cause was MMAE toxicity. Furthermore, it should be noted that there was variability in the growth rate of PDX tumors, with some tumors growing much more slowly than others. However, even when all animals were included in the analysis, a statistically significant difference was observed.

[0174] The safety of CTT2274 was compared to that of MMAE at the same dose in mice. As previously mentioned, MMAE ADCs have many known associated toxicities. Furthermore, given the mechanism of action of MMAEs, organs and tissues with high cell turnover rates (e.g., bone marrow, gastrointestinal tract, spleen, thymus) are likely to be adversely affected by MMAE-based treatment. In addition, PSMA is expressed in healthy proximal tubular epithelial cells of the kidney, albeit at lower levels than in tumor cells. Although PSMA is expressed in proximal tubular cells, these cells also express MDR1, known as an MMAE efflux transporter. Therefore, CTT2274 may cause kidney damage.

[0175] While CTT2274 showed promising efficacy, survival, and safety results, this trial is not without its limitations. As with all animal models of cancer, the subcutaneous PDX PCa model used does not fully reflect the human condition. Humans diagnosed with PCa have a localized disease and often metastasize to bones, lungs, liver, and other tissues, a phenomenon that this model cannot replicate. Future trials aim to clarify the pharmacokinetics and complete safety / toxicity profile of CTT2274.

[0176] This study demonstrated that CTT2274 is a safe and effective treatment for PCa. Adding a biphenyl motif to CTT2274 significantly improved treatment duration and overall survival compared to CTT2101 without the biphenyl motif. Furthermore, CTT2274 was shown to be at least as effective as, and in some cases more effective than, MMAE. Additionally, treatment with CTT2274 resulted in at least equivalent or improved overall survival compared to MMAE. Finally, CTT2274 demonstrated a superior safety / toxicity profile compared to MMAE. These results support the need for additional in vivo studies to evaluate the clinical applicability of CTT2274.

[0177] Example 10. Dose-range efficacy test This study was conducted at The Jackson Laboratory in Sacramento, California, using a TM00298 patient-derived xenograft (PDX) prostate cancer tumor model. Male NSG mice (stock #005557) aged 6-8 weeks were subcutaneously inoculated with 40 μL of tumor tissue into the right flank. Based on tumor volume, mice were randomly assigned to either the CTT2274 or PBS treatment group, and t-tests were performed to confirm no statistically significant difference between the groups. 56 mice (n=8 / group) were administered either 100 μL of 1X PBS or 100 μL of 1X PBS containing 0.36 mg / kg, 0.72 mg / kg, 1.44 mg / kg, 3.6 mg / kg, 9 mg / kg, or 18 mg / kg of CTT2274 via tail vein injection. Mice received the treatment once a week for 6 weeks, starting from day 0. During the study period, animals were observed daily for signs of illness and death. Weight was measured twice a week. Tumor volume was measured twice a week using calipers (width squared × 0.5 × length). The tumor volume was 2,000 mm³. 3 Animals were euthanized if the following conditions were exceeded, if the body condition score (BCS) was 2 or less, or if the tumor was found to be ulcerated or necrotic. This study was conducted blindly.

[0178] result At the start of the study, the body weights of mice in the 0.36 mg / kg, 0.72 mg / kg, 1.44 mg / kg, 3.6 mg / kg, 9 mg / kg, 18 mg / kg, and PBS groups were 31.26 ± 1.92 g, 30.94 ± 2.55 g, 31.19 ± 1.74 g, 29.98 ± 2.27 g, 31.16 ± 1.80 g, 31.49 ± 2.03 g, and 30.91 ± 1.22 g groups, respectively (Figure 9A). In the 18 mg / kg group, the first weight loss was observed on day 17 of the study, and further weight loss was observed on day 21. In the 9 mg / kg group, weight loss was observed from day 52 of the study, and the animals' body weight fluctuated throughout the remainder of the study. No such decreases or fluctuations were observed in the 0.36 mg / kg, 0.72 mg / kg, 1.44 mg / kg, and 3.6 mg / kg groups.

[0179] The tumor volume at the start of the study was 54.47–234.54 mm² for all subjects. 3 The mean tumor volume before the first dose was 111.94 ± 42.45 mm for the 0.36 mg / kg group, 0.72 mg / kg group, 1.44 mg / kg group, 3.6 mg / kg group, 9 mg / kg group, 18 mg / kg group, and PBS group, respectively. 3 , 112.14±46.54mm 3 , 111.76±44.36mm 3 , 111.53±50.88mm 3 , 112.05±51.58mm 3 , 112.24±61.48mm 3 , 111.88±55.24mm 3(Figures 9B and 9C). A dose-response trend was observed in tumor suppression. Figure 10 shows the tumor growth trends between the mouse groups. This data shows that when doses of 0.36 mg / kg and 0.72 mg / kg were administered once a week for 6 weeks, there was little effect on suppressing tumor growth. At a dose of 1.44 mg / kg, tumor suppression improved with the same regimen. Furthermore, doses of 3.6 mg / kg and 9 mg / kg in the same regimen were very effective in suppressing tumor growth, and the tumors did not appear to reach the exponential growth phase until day 84 of the study, indicating that tumor suppression persisted for 7 weeks after the end of administration. Interestingly, mice #07 and #22 in the 9 mg / kg group showed no measurable tumors from day 28 and day 80 of the study, respectively. Mouse #79 in the 9 mg / kg group showed no measurable tumors from days 45 to 84 of the study, but a tumor became measurable on day 87.

[0180] The median survival time was 38 days, 47 days, 70 days, 21 days, and 40 days for the 0.36 mg / kg, 0.72 mg / kg, 1.44 mg / kg, 18 mg / kg, and PBS groups, respectively (Figure 9D). All mice in the 18 mg / kg group were euthanized on day 21 of the study at the recommendation of the veterinarian due to gait abnormalities. All mice in the 0.36 mg / kg, 0.72 mg / kg, and PBS groups were euthanized because they met the euthanasia criteria for tumor volume. Mice in the 1.44 mg / kg group that did not complete the study were euthanized because they met the euthanasia criteria for tumor volume. Mice in the 9 mg / kg group that did not complete the study were euthanized because their BCS was 2. In addition, one mouse exhibited respiratory / wheezing abnormalities due to an unexplained tumor above the heart compressing the trachea. At the end of the experiment (day 90), all mice in the 3.6 mg / kg group, 6 mice in the 9 mg / kg group, and 2 mice in the 1.44 mg / kg group were still alive. Table 2 shows a comparison of survival statistics between the groups. [Table 2]

[0181] Since gait abnormalities were observed in the 18 mg / kg group, two mice were randomly selected, euthanized, fixed in formalin, and subjected to macroscopic necropsy and histopathological examination. In both mice, the clinical signs and gait abnormalities were attributed to axonal degeneration and neuronal degeneration affecting the dorsal root ganglia and ventral white matter of the spinal cord.

[0182] Based on the above-described administration studies in mice, it is expected that safe and effective dose ranges for compounds pursuant to this disclosure, including, for example, CTT2274, can be achieved within a physiologically safe, effective, and / or tolerable range (as suggested, for example, by the efficacy in mice receiving doses in the range of 1.44–9 mg / kg).

[0183] Examples 11-34. Additional SMDCs according to the present disclosure. Additional exemplary SMDCs are prepared substantially according to the procedure described above and are shown in Table 3. In particular, the azide reacts with the target molecule to produce the corresponding reaction product, namely the triazole-containing SMDC. Thus, the exemplary compounds in Table 3 are reaction products of azide and alkyne. For each example in Table 3, the target molecule (i.e., alkyne) is CTT2270, CTT23002, CTT23012, CTT23022, CTT23032, CTT23042, or CTT23052, and their structures are as follows: [ka] [ka]

[0184] Further examples of SMDC precursors are prepared substantially according to the procedure described above and are also shown in Table 3. In particular, reacting an azide with an alkyne containing a carboxylic acid yields the corresponding reaction product, namely a triazole-containing SMDC precursor. Each example of the triazole-containing SMDC precursor in Table 3 can be further derivatized at the target moiety using the carboxylic acid moiety (e.g., by various amide bond formation reactions). In each example in Table 3, the target molecule (i.e., alkyne) is DBCO-Bip, whose structure is as follows: [ka] [Table 3-1] [Table 3-2] [Table 3-3]

[0185] The examples and embodiments described herein are for illustrative purposes only, and it is understood that various modifications or changes are proposed in light of them and incorporated into the spirit and scope of this application and the appended claims. All publications, patents, and patent applications referenced herein are incorporated herein by reference for all purposes.

Claims

1. Compound of structural formula (I) 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, in the formula, D is -NR ２ A therapeutic or diagnostic agent bound to L via a -, -S-, or -O- moiety, L is an acid-cleavable linker, which combines to provide a carbamate moiety in formula (I). Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R ２ It forms an amide with nitrogen having the following properties: m is an integer from 1 to 5, n is an integer from 1 to 4, and R １ , R ２ , R ３ , and R ４ However, H and C are independent of each other. １ -C ６ The present invention provides compound (I), which is an alkyl group or a protecting group, or a pharmaceutically acceptable salt thereof.

2. formula 【Chemistry 2】 A compound according to claim 1, wherein the formula is D is -NR ２ -, a therapeutic agent or diagnostic agent bound to L via a -S- or -O- moiety, L is an acid-cleavable linker, which combines to provide a carbamate moiety in formula (I). Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R ２ It forms an amide with nitrogen having the following properties: m is an integer from 1 to 5, n is an integer from 1 to 4, and R １ , R ２ , R ３ , and R ４ However, H and C are independent of each other. １ -C ６ The present invention provides compound (I), which is an alkyl group or a protecting group, or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1 or claim 2, wherein D is a therapeutic agent.

4. A compound according to any one of claims 1 to 3, wherein D is an anti-angiogenic agent, a cytotoxic agent, a cytokine, a chemokine, an apoptotic agent, a prodrug, a toxin, an enzyme, a radioisotope, an immunomodulator, an antibiotic, a drug active in the CNS, or a hormone.

5. D is monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), exatecan, N-Me-L-Ala-mytansinol, gemcitabine, seco-duocarmycin (seco-DUBA), gemcitabine monophosphate, doxorubicin, cabazitaxel, docetaxel, paclitaxel, imiquimod, 7-ethyl-10-hydroxycamptothecin (SN-38), duocarmycin (DUBA), or (R)-5-chloro-N ２ -[4-(4-methylpiperazine-1-yl)phenyl]-N ４ A compound according to any one of claims 1 to 3, selected from -[(tetrahydrofuran-2-yl)methyl]pyrimidine-2,4-diamine((R)-9b).

6. The compound according to claim 1 or claim 2, wherein D is a diagnostic agent.

7. A compound according to claim 1 or claim 2, wherein D is a radioisotope, an imaging agent, a fluorescent dye, a near-infrared dye, an enzyme, a chemiluminescent agent, a bioluminescent agent, a paramagnetic ion, an ultrasonic label, or a radioacoustic label.

8. D is -NR ２ The compound according to any one of claims 1 to 7, which is bonded to L via a -.

9. The compound according to any one of claims 1 to 7, wherein D is bonded to L via -O-.

10. A compound according to any one of claims 1 to 7, wherein L is 【Transformation 3】 Selected from, In the formula, R ５ However, H, -OH, or C １ -C ６ A compound that is an alkoxyl.

11. A compound according to any one of claims 1 to 7, wherein L is 【Chemistry 4】 Selected from, in the formula, R ５ However, H, -OH, or C １ -C ６ A compound that is an alkoxyl.

12. R ５ The compound according to claim 10 or 11, wherein is H.

13. R ５ ga-OCH ３ The compound according to claim 10 or 11.

14. A compound according to any one of claims 1 to 7, wherein L is 【Transformation 5】 It is a compound.

15. A compound according to any one of claims 1 to 14, wherein the N-terminus of X is bonded to an adjacent carbonyl to provide an amide moiety.

16. The compound according to any one of claims 1 to 15, wherein each X is independently selected from naturally occurring amino acids (e.g., L-amino acids) or unnaturally occurring amino acids (e.g., D-amino acids).

17. A compound according to any one of claims 1 to 16, wherein at least one X is structural 【Transformation 6】 It has, in the formula, each R ６ These are independent of each other: (i) H, (ii) C １ -C ６ Alkyl, (iii) hydroxy(C) １ -C ６ (Alkyl), (iv) C １ -C ６ Alkoxy (C １ -C ６ Alkyl), (v)C ２ -C ６ Alkenil, (vi)R ８ R ９ N(C) １ -C ６ Alkyl), (vii) Halogen, (viiii) Halo (C) １ -C ６ Alkyl), (ix)carboxy(C １ -C ６ Alkyl), (x) thio (C １ -C ６ Alkyl), (xi)C １ -C ６ Alkylthio (C １ -C ６ Alkyl), (xi)R ８ R ９ NC(O)(C １ -C ６ Alkyl), (xiiii)C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), (xiv)C ３ -C ８ Heterocyclyl (C １ -C ６ ) alkyl, (xv)aryl (C １ -C ６ Alkyl), (xvi) heteroaryl C １ -C ６ (alkyl), and (xvii)R １７ C(O)NR ８ (C １ -C ６ Selected from alkyl, R ８ and R ９ are each independently selected from hydrogen and C １ -C ６ alkyl (for example, C １ -C ４ alkyl, or C １ -C ３ alkyl), R １７ is C １ -C ６ alkyl, hydroxy(C １ -C ６ alkyl), C １ -C ６ alkoxy(C １ -C ６ alkyl), C ２ -C ６ alkenyl, -NR １０ R １１ , R １０ R １１ N(C １ -C ６ alkyl), C ３ -C １０ cycloalkyl(C １ -C ６ alkyl), C ３ -C ８ heterocyclyl(C １ -C ６ alkyl), aryl(C １ -C ６ alkyl), heteroaryl(C １ -C ６ alkyl), C ３ -C １０ cycloalkyl, C ３ -C ８ heterocyclyl, aryl, or heteroaryl, Each cycloalkyl, heterocyclyl, aryl, and heteroaryl can optionally be -OH, -SH, or -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Substituted with alkyl, R １０ and R １１ However, independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

18. A compound according to any one of claims 1 to 16, wherein at least one X is structural 【Transformation 7】 It has, A compound according to claim 1, wherein the formula is p is an integer from 1 to 5, q is an integer from 1 to 5, Each R ７ However, each is independent of H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl, C ３ -C ８ Heterocyclyl, aryl, heteroaryl, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Selected from alkyl, Each R １７ However, each is independent of C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), heteroaryl (C １ -C ６ Alkyl), C ３ -C １０ Cycloalkyl, C ３ -C ８ Selected from heterocyclyl, aryl, and heteroaryl compounds, Each cycloalkyl, heterocyclyl, aryl, and heteroaryl can optionally be -OH, -SH, or -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Substituted with alkyl, each R １０ and R １１ However, independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

19. The compound according to claim 18, wherein p is 1.

20. The compound according to claim 18 or claim 19, wherein q is 1.

21. p is 1, q is 1, R ７ The compound according to claim 18, wherein is an aryl (for example, phenyl).

22. A compound according to any one of claims 1 to 16, wherein at least one X is structural 【Transformation 8】 A compound according to claim 1, wherein the formula is r is an integer from 1 to 5, s is an integer between 1 and 7, and Each R １２ However, each is independent of H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl, C ３ -C ８ Heterocyclyl, aryl, heteroaryl, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Selected from alkyl groups, Each cycloalkyl, heterocyclyl, aryl, and heteroaryl can optionally be -OH, -SH, or -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Substituted with alkyl, each R １０ and R １１ However, independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

23. The compound according to claim 22, wherein r is 1.

24. The compound according to claim 22 or claim 23, wherein s is 1.

25. The compound according to claim 18, wherein at least one X is structural 【Chemistry 9】 A compound according to claim 1, wherein the formula is a is an integer from 1 to 4, b is an integer between 1 and 5, and R １３ and R １４ Each of these is independently H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Selected from alkyl, and halogen, R １０ and R １１ However, each independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

26. The compound according to claim 22, wherein at least one X is structural 【Chemistry 10】 A compound according to claim 1, wherein the formula is c is an integer between 1 and 3. d is an integer from 1 to 4, and, R １５ and R １６ Each of these is independently H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Selected from alkyl, and halogen, R １０ and R １１ However, each independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

27. The compound according to any one of claims 1 to 26, wherein n is an integer from 1 to 3.

28. The compound according to any one of claims 1 to 26, wherein n is an integer of 1 or 2.

29. The compound according to any one of claims 1 to 26, wherein n is 1.

30. The compound according to any one of claims 1 to 26, wherein n is 1 and X is phenylalanine.

31. A compound according to any one of claims 1 to 26, wherein n is 1 and X is 【Chemistry 11】 It is a compound.

32. A compound according to any one of claims 1 to 26, wherein n is 1 and X is 【Chemistry 12】 It is a compound.

33. A compound according to any one of claims 1 to 32, wherein m is an integer from 1 to 4 (for example, an integer from 1 to 3).

34. The compound according to any one of claims 1 to 32, wherein m is 1 or 2.

35. A compound according to any one of claims 1 to 32, wherein m is 1.

36. A compound according to any one of claims 1 to 35, R １ , R ２ , and R ３ However, independently, H or C １ -C ４ A compound that is alkyl.

37. A compound according to any one of claims 1 to 35, R １ , R ２ , and R ３ A compound in which each element is independently either H or methyl.

38. A compound according to any one of claims 1 to 35, R １ , R ２ , and R ３ However, each of these compounds is independently a hydrogen atom.

39. R ４ The compound according to any one of claims 1 to 35, wherein is H or methyl.

40. R ４ The compound according to any one of claims 1 to 35, wherein is H.

41. A compound according to any one of claims 1 to 40, of the following formula. 【Chemistry 13】

42. The compound according to claim 41, wherein D is MMAE.

43. A compound according to any one of claims 1 to 40, the formula 【Chemistry 14】 A compound of the same name.

44. A pharmaceutical composition comprising a compound according to any one of claims 1 to 43, and a pharmaceutically acceptable excipient, carrier, adjuvant, stabilizer, and / or diluent.

45. A method for delivering a therapeutic agent or diagnostic agent to a target, wherein the method comprises administering a therapeutically effective amount of a compound according to any one of claims 1 to 43 or a pharmaceutical composition according to claim 44 to a target requiring such agent, wherein D is a therapeutic agent or diagnostic agent.

46. A method for treating a patient with prostate cancer, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 43 or a pharmaceutical composition according to claim 44.

47. A method for imaging one or more prostate cancer cells in a patient, comprising administering to the patient a compound according to any one of claims 1 to 43 or a pharmaceutical composition according to claim 44.

48. Compound of structural formula (II) 【Chemistry 15】 or a pharmaceutically acceptable salt thereof, in the formula, Each Y is independently a natural or unnatural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety. m is an integer from 1 to 5, and A compound or a pharmaceutically acceptable salt thereof, where n is an integer between 1 and 4.

49. A compound according to claim 48, wherein each Y is independently selected from naturally occurring amino acids (e.g., L-amino acids) or unnaturally occurring amino acids (e.g., D-amino acids).

50. A compound according to claim 48 or claim 49, wherein at least one Y is structural 【Chemistry 16】 It has, in the formula, each R ６ These are (i) H and (iii) C, respectively, independently. １ -C ６ Alkyl, (iii)hydroxy(C) １ -C ６ (Alkyl), (iv) C １ -C ６ Alkoxy (C １ -C ６ Alkyl), (v)C ２ -C ６ Alkenil, (vi)R ８ R ９ N(C) １ -C ６ Alkyl), (vii) Halogen, (viiii) Halo (C) １ -C ６ Alkyl), (ix)carboxy(C １ -C ６ Alkyl), (x) thio (C １ -C ６ Alkyl), (xi)C １ -C ６ Alkylthio(C １ -C ６ Alkyl), (xi)R ８ R ９ NC(O)(C １ -C ６ Alkyl), (xiiii)C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), (xiv)C ３ -C ８ Heterocyclyl (C １ -C ６ ) alkyl, (xv)aryl (C １ -C ６ Alkyl), (xvi) heteroaryl (C １ -C ６ Alkyl), (xvii)R １７ C(O)NR ８ (C １ -C ６ Alkyl), (xviiii)C ３ -C １０ Cycloalkyl, (xix)C ３ -C ８ Selected from heterocyclyl, (xx)aryl, and (xvi) heteroaryl, R ８ and R ９ However, each is independent of hydrogen and C １ -C ６ Alkyl (for example, C １ -C ４ Alkyl, or C １ -C ３ Selected from alkyl, R １７ However, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), heteroaryl (C １ -C ６ Alkyl), C ３ -C １０ Cycloalkyl, C ３ -C ８ It is a heterocyclyl, aryl, or heteroaryl compound. Each cycloalkyl, heterocyclyl, aryl, and heteroaryl can optionally be -OH, -SH, or -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ It is substituted with alkyl, where R １０ and R １１ However, independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

51. A compound according to claim 48 or claim 49, wherein at least one Y is structural 【Chemistry 17】 It has, A compound according to claim 1, wherein the formula is p is an integer from 1 to 5, q is an integer from 1 to 5, and Each R ７ However, each is independent of H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl, C ３ -C ８ Heterocyclyl, aryl, heteroaryl, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Selected from alkyl groups, Each R １７ However, each is independent of C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), heteroaryl (C １ -C ６ Alkyl), C ３ -C １０ Cycloalkyl, C ３ -C ８ Selected from heterocyclyl, aryl, and heteroaryl compounds, Each cycloalkyl, heterocyclyl, aryl, and heteroaryl can optionally be -OH, -SH, or -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Substituted with alkyl, each R １０ and R １１ However, independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

52. The compound according to claim 51, wherein p is 1.

53. The compound according to claim 51 or claim 52, wherein q is 1.

54. The compound according to claim 51, wherein p is 1, q is 1, and R ７ A compound in which the parent element is aryl (for example, phenyl).

55. A compound according to claim 48 or claim 49, wherein at least one Y is [Chemistry 18] A compound according to claim 1, wherein the formula is r is an integer from 1 to 5, s is an integer between 1 and 7, and Each R １２ However, each is independent of H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl, C ３ -C ８ Heterocyclyl, aryl, heteroaryl, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Selected from alkyl groups, Each cycloalkyl, heterocyclyl, aryl, and heteroaryl can optionally be -OH, -SH, or -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Alkyl), halogen, C ３ -C １０ Cycloalkyl (C １ -C ６ Alkyl), C ３ -C ８ Heterocyclyl (C １ -C ６ Alkyl), aryl (C １ -C ６ Alkyl), or heteroaryl (C １ -C ６ Substituted with alkyl, R １０ and R １１ However, independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

56. The compound according to claim 55, wherein r is 1.

57. The compound according to claim 55 or claim 56, wherein s is 1.

58. The compound according to claim 51, wherein at least one Y is structural 【Chemistry 19】 A compound according to claim 1, wherein the formula is a is an integer from 1 to 4, b is an integer between 1 and 5, and R １３ and R １４ Each of these is independently H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Selected from alkyl, and halogen, R １０ and R １１ However, each independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

59. The compound according to claim 55, wherein at least one Y is 【Chemistry 20】 A compound according to claim 1, wherein the formula is c is an integer between 1 and 3. d is an integer from 1 to 4, and, R １５ and R １６ Each of these is independently H, -OH, -SH, and -NO. ２ , -COOH, C １ -C ６ Alkyl, hydroxy (C １ -C ６ Alkyl), C １ -C ６ Alkoxy (C １ -C ６ Alkyl), C ２ -C ６ Alkenyl, -NR １０ R １１ , R １０ R １１ N(C) １ -C ６ Selected from alkyl, and halogen, R １０ and R １１ However, each independently, hydrogen and C １ -C ６ A compound selected from alkyl groups.

60. A compound according to any one of claims 48 to 59, wherein n is in the range of 1 to 3.

61. A compound according to any one of claims 48 to 59, wherein is 1 or 2.

62. The compound according to any one of claims 48 to 59, wherein n is 1.

63. The compound according to any one of claims 48 to 59, wherein n is 1 and Y is phenylalanine.

64. A compound according to any one of claims 48 to 59, wherein n is 1 and Y is 【Chemistry 21】 It is a compound.

65. A compound according to any one of claims 48 to 59, wherein n is 1 and Y is 【Chemistry 22】 It is a compound.

66. The compound according to any one of claims 48 to 65, wherein m is in the range of 1 to 4 (for example, in the range of 1 to 3).

67. The compound according to any one of claims 48 to 65, wherein m is 1 or 2.

68. The compound according to any one of claims 48 to 65, wherein m is 1.

69. Compound of formula (III) 【Chemistry 23】 or a pharmaceutically acceptable salt thereof, in the formula, Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R ２ It forms an amide with nitrogen having the following properties: m is an integer from 1 to 5, n is an integer from 1 to 4, and R １ , R ２ , R ３ , and R ４ However, H and C are independent of each other. １ -C ６ The present invention provides compound (I), which is an alkyl group or a protecting group, or a pharmaceutically acceptable salt thereof.

70. A compound according to claim 68, the formula 【Chemistry 24】 A compound according to claim 1, wherein the formula is Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R ２ It forms an amide with nitrogen having the following properties: m is an integer from 1 to 5, n is an integer from 1 to 4, and R １ , R ２ , R ３ , and R ４ However, H and C are independent of each other. １ -C ６ The present invention provides compound (I), which is an alkyl group or a protecting group, or a pharmaceutically acceptable salt thereof.

71. A compound according to claim 69 or claim 70, wherein X is as defined in any one of claims 15 to 26.

72. A compound according to any one of claims 69 to 71, wherein n is defined in the same manner as in any one of claims 27 to 32.

73. A compound according to any one of claims 69 to 72, wherein m is defined in the same manner as in any one of claims 33 to 35.

74. A compound according to any one of claims 69 to 73, R １ , R ２ , R ３ , and R ４ A compound, defined in the same manner as any one of claims 36 to 40.

75. Compound of formula (IV) 【Chemistry 25】 or a pharmaceutically acceptable salt thereof, in the formula, D is -NR ２ A therapeutic or diagnostic agent bound to L via a -, -S-, or -O- moiety, L is an acid-cleavable linker, which combines to provide a carbamate moiety in formula (IV). Each Y is independently a natural or unnatural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety. m is an integer from 1 to 5, and n is an integer between 1 and 4, which is a compound or a pharmaceutically acceptable salt thereof.

76. A compound according to claim 75, wherein D is as defined in any one of claims 3 to 9.

77. A compound according to claim 75 or claim 76, wherein L is as defined in any one of claims 10 to 14.

78. A compound according to any one of claims 75 to 77, wherein Y is defined in any one of claims 48 to 59.

79. A compound according to any one of claims 75 to 78, wherein n is defined in any one of claims 60 to 65.

80. A compound according to any one of claims 75 to 79, wherein m is defined in any one of claims 66 to 68.

81. A compound according to any one of claims 75 to 80, R １ , R ２ , R ３ , and R ４ A compound, defined in the same manner as any one of claims 36 to 40.

82. A method for synthesizing a compound of formula (I) according to any one of claims 1 to 43, wherein the compound of formula (V) 【Chemistry 26】 or a pharmaceutically acceptable salt thereof Compounds of formula (III) (for example, those described in any one of claims 69 to 74) 【Chemistry 27】 or a pharmaceutically acceptable salt thereof, comprising contacting with a pharmaceutically acceptable salt thereof in the formula Each X is independently a natural or unnatural n α-amino acid, where the C-terminus of X is R ２ It forms an amide with nitrogen having the following properties: m is an integer from 1 to 5, n is an integer from 1 to 4, and R １ , R ２ , R ３ , and R ４ However, H and C are independent of each other. １ -C ６ The present invention provides compound (I), which is an alkyl group or a protecting group, or a pharmaceutically acceptable salt thereof.

83. A method for synthesizing a compound of formula (IV) according to any one of claims 75 to 81, wherein the compound of formula (V) 【Chemistry 28】 or a pharmaceutically acceptable salt thereof Compounds of formula (II) (for example, those described in any one of claims 48 to 68) 【Chemistry 29】 or a pharmaceutically acceptable salt thereof, comprising contacting with a pharmaceutically acceptable salt thereof in the formula Each Y is independently a natural or unnatural n α-amino acid, where the N-terminus of Y is bonded to an adjacent carbonyl to provide an amide moiety. m is an integer from 1 to 5, and n is an integer between 1 and 4.

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