WO2024199473A1

WO2024199473A1 - Methods and compositions for targeted protein degradation

Info

Publication number: WO2024199473A1
Application number: PCT/CN2024/084926
Authority: WO
Inventors: Wei Yin; Weiwen Ying; Kevin Paul Foley; Yan Dai; Xiangcai YANG; Guoqiang Wang; Thomas PRINCE
Original assignee: Ranok Therapeutics Hangzhou Co Ltd
Current assignee: Ranok Therapeutics Hangzhou Co Ltd
Priority date: 2023-03-31
Filing date: 2024-03-29
Publication date: 2024-10-03
Anticipated expiration: 2025-09-30

Abstract

Provided are chaperone retention effect drug conjugates (CREDCs) comprised of an exatecan-like portion, a linker, and a chemical moiety that binds HSP90 protein. Pharmaceutical compositions comprising the disclosed CREDCs and their use in treating cancer and related conditions are also provided.

Description

METHODS AND COMPOSITIONS FOR TARGETED PROTEIN DEGRADATION

RELATED APPLICATIONS

This application claims the benefit of priority to international application No. PCT/CN2023/085328, filed March 31, 2023, the entire contents of which are incorporated herein by reference.

BACKGROUND

Despite advances, treatment of cancer with chemotherapeutic agents remains disappointing. Cancer patients treated with chemotherapeutic agents endure debilitating regimens and often a poor prognosis. This is because the effectiveness and/or application of chemotherapeutic agents containing toxic moieties is reduced by unwanted side effects due to off target or systemic toxicity. The ability to concentrate chemotherapeutics agents in tumor tissue while reducing normal tissue exposure may greatly reduce off target toxicity and improve overall clinical outcomes.

Cancer is characterized by aberrant protein expression and activity. HSP90 (heat shock protein 90 kDa) is a molecular chaperone that facilitates the 3-dimensional folding and maintenance of a significant portion of the proteins that signal for cellular growth including many oncogenes (Trepel J. et al, Nat Rev Cancer. 2010, 10: 537-49; Birbo B. et al, Int J Mol Sci., 2021, 22: 10317) . Cancer cells exist in tumor microenvironments characterized by reduced blood flow, high interstitial pressure, hypoxia and acidosis. To mitigate these conditions that negatively affect overall protein stability, cancer cells overexpress HSP90 and even secrete it extracellularly (Lang BJ et al, Int J Mol Sci. 2019, 20: 4507; Taha EA et al, Int J Mol Sci. 2019, 20: 4588) . Moreover, HSP90 chaperone activity is greatly enhanced in cancer cells due to the abundance of mutated or misfolded proteins (Lacey T, Lacey H, Cancer Treat Res Commun. 2021, 28: 100400) . Cancer specific post-translational modification and protein-protein interaction states also affect HSP90 chaperone activity (Backe SJ et al, J Biol Chem. 2020, 295: 11099-11117) . In cancer cells this elevated expression and enhanced chaperone activity results in increased binding of HSP90 to small molecule drugs that target its N-terminal ATP-binding. This is in contrast to normal, non-proliferating cells that do not possess elevated levels of HSP90 expression or activity (Kamal A et al., Nature. 2003 425: 407-10; Moulick K et al, Nat Chem Biol. 2011, 7: 818-26) . Due to these differences, HSP90 drugs exhibit a chaperone retention effect by concentrating in tumor tissue, while cleared from normal tissue.

Camptothecin analogs such as irinotecan, exatecan and SN-38 are antineoplastic agents that inhibit DNA topoisomerase I (Topo I) activity and prevent the relaxation of supercoiled DNA during DNA replication and RNA transcription (Pommier Y, Nat Rev Cancer. 2006, 6: 789–802) . These agents are commonly used in chemotherapeutic regimen to treat colorectal, pancreatic, small lung and gastric cancers along with pediatric sarcomas (Fujita K et al, World J Gastroenterol. 2015, 21: 12234–12248; Rubinson DA et al, Hematol Oncol Clin North Am. 2015, 29: 761–776; Levy B et al, JNCCN. 2013, 11: 780–787; Mahipal A et al, JNCCN. 2015, 13: 1281–1291; Yohe ME et al, Cancer Treat Res Commun. 2021, 28: 100400) . However, these agents present problems with bioavailability and off target toxicity (Venditto VJ, Simanek ES, Mol Pharm. 2010, 7: 307-49) . Accordingly, the need exists for chaperone retention effect drug conjugates (CREDCs) that effectively target and concentrate antineoplastic agents such as camptothecin analogs into tumor tissue.

SUMMARY

Provided herein are CREDCs comprising an exatecan-like portion, a linker, and a chemical moiety that binds HSP90 protein. Such compounds include those having the Formula I:

and pharmaceutically acceptable salts thereof, wherein R^Q, R^T, X, L, and A are as defined herein. Compositions comprising the disclosed compounds of Formula I as well as methods for their manufacture and their biochemical activity are also provided.

DETAILED DESCRIPTION

1. General Description of Compounds

Provided herein are CREDC compounds having the Formula I:

or a pharmaceutically acceptable salt thereof, wherein

R^Q and R^T are each independently selected from H, optionally substituted (C₁-C₄) alkyl, optionally substituted aryl, and optionally substituted heteroaryl, or R^Q and R^T are taken together with the carbon they are attached to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl;

X is absent or -NR^W, wherein R^W is H or (C₁-C₄) alkyl;

L is a linker; and

A is a chemical moiety that binds HSP90 protein

2. Definitions

As used herein, the articles “a” and “an” refer to one or more than one, e.g., to at least one, of the grammatical object of the article. The use of the words "a" or "an" when used in conjunction with the term "comprising" herein may mean "one, " but it is also consistent with the meaning of "one or more, " "at least one, " and "one or more than one. "

As used herein, “about” and “approximately” generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%) , typically, within 10%, and more typically, within 5%of a given range of values. The term “substantially” means more than 50%, preferably more than 80%, and most preferably more than 90%or 95%.

As used herein the term "comprising" or "comprises" are used in reference to compositions, methods, and respective component (s) thereof, that are present in a given embodiment, yet open to the inclusion of unspecified elements.

As used herein, the term "alkyl" means a saturated straight chain or branched non-cyclic hydrocarbon having, unless specified otherwise, from 1 to 10 carbon atom e.g., (C₁-C₆) alkyl or (C₁-C₄) alkyl. Representative straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl and n-decyl; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylbutyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2, 2-dimethylpentyl, 2, 2-dimethylhexyl, 3, 3-dimethylpentyl, 3, 3-dimethylhexyl, 4, 4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2, 2-diethylpentyl, 3, 3-diethylhexyl, 2, 2-diethylhexyl, 3, 3-diethylhexyl and the like.

As used herein, the term “cycloalkyl" means a saturated, monocyclic alkyl radical having from e.g., 3 to 10 carbon atoms (e.g., from 4 to 6 carbon atoms) . Representative cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecanyl.

The term “oxo” refers to the group =O.

As used herein, the term "haloalkyl" means and alkyl group in which one or more (including all) the hydrogen radicals are replaced by a halo group, wherein each halo group is independently selected from -F, -Cl, -Br, and -I. Representative haloalkyl groups include trifluoromethyl, bromomethyl, 1, 2-dichloroethyl, 4-iodobutyl, 2-fluoropentyl, and the like.

“Alkoxy” means an alkyl radical attached through an oxygen linking atom, represented by –O-alkyl. For example, “ (C₁-C₄) alkoxy” includes methoxy, ethoxy, proproxy, and butoxy.

“Haloalkoxy” is a haloalkyl group which is attached to another moiety via an oxygen atom such as, e.g., –OCHF₂ or –OCF₃.

As used herein, the term "heterocyclyl" means a 4-to 12-membered monocyclic or polycyclic saturated or partially unsaturated heterocyclic ring (e.g., bridged bicyclic) containing 1 to 4 heteroatoms independently selected from N, O, and S. The heterocycle may be attached via any heteroatom or carbon atom, as valency permits. Representative heterocycles include morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, oxiranyl, dioxanyl, oxetanyl, dihydrofuranyl, dihydropyranyl, isoindolinyl, dihydropyridinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl, tetrahydropyrimidinyl, diazabicyclooctanyl, hexahydropyrrolizinyl, 2-azaspiro [3.3] heptanyl, 2, 7-diazaspiro [3.5] nonanyl, 2-azaspiro [3.5] nonanyl, 3-azabicyclo [3.1.0] hexanyl, 8-azabicyclo [3.2.1] octanyl, 3, 8-diazabicyclo [3.2.1] octanyl, 3, 6-diazabicyclo [3.1.1] heptanyl, octahydro-1H-pyrrolo [2, 3-c] pyridinyl, and the like. Optional substituents on a heterocyclyl group may be present on any substitutable position and, include, e.g., the position at which the heterocyclyl is attached, valence permitting.

As used herein, the term "heteroaryl" means a 5-to 12-membered aromatic radical containing 1-4 heteroatoms selected from N, O, and S. A heteroaryl group may be mono-or bicyclic. The heteroaryl may be attached via any heteroatom or carbon atom, as valency permits. Representative heteroaryl groups include pyridyl, furanyl, thienyl, pyrrolyl, oxazolyl, imidazolyl, thiazolyl, isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, triazolyl, thiadiazolyl, isoquinolinyl, indazolyl, benzoxazolyl, benzofuryl, indolizinyl, imidazopyridyl, tetrazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl, azaindolyl, imidazopyridyl, quinazolinyl, purinyl, benzothienyl, and the like. Optional substituents on a heteroaryl group may be present on any substitutable position and, include, e.g., the position at which the heteroaryl is attached, valence permitting.

As used herein, the term "halogen" or "halo" means F, Cl, Br or I.

The term "linker" or "tether, " used interchangeably, refers to a chemical moiety that joins two other moieties (e.g., a therapeutic moiety and a binding moiety) . A linker can covalently join a therapeutic moiety (e.g., exatecan) and a binding moiety (e.g., chemical moiety which binds HSP90 protein. In one aspect, the linker is uncleavable in vivo. In one aspect, the linker comprises one or more cyclic ring systems. In another aspect, the linker comprises an alkyl chain optionally substituted by and/or interrupted with one or more chemical groups. In one aspect, the linker comprises optimal spatial and chemical properties to effectuate optimal therapeutic activity.

The term “HSP90” refers collectively, individually or in various combinations to the protein products of members of the heat shock protein 90 (90 kDa) gene family, including: HSP90AA1 (HSP90-alpha or HSP90α) , HSP90AB1 (HSP90-beta or HSP90β) , HSP90B1 (GRP94) and TRAP1.

When used in connection to describe a chemical group that may have multiple points of attachment, a hyphen (-) designates the point of attachment of that group to the variable to which it is defined. For example, -NH (C₁-C₄) alkyl _\means that the point of attachment for this group occurs on the nitrogen atom.

A hash bond as inrepresents the point at which the depicted group is attached to the defined variable.

The compounds described herein may have chiral centers and/or geometric centers (E-and Z-isomers) . It will be understood that the present disclosure encompasses all stereoisomers and geometric isomers. Tautomeric forms of the compounds described herein are also part of the present disclosure.

When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%or 99.9%by weight pure relative to all of the other stereoisomers. Percent by weight pure relative to all of the other stereoisomers is the ratio of the weight of one stereoisomer over the weight of the depicted stereoisomer plus the weight of the other stereoisomers.

For use in medicines, the pharmaceutically acceptable salts of the disclosed compounds refer to non-toxic “pharmaceutically acceptable salts. ” Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic/anionic or basic/cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include e.g., salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, nitric, and sulfuric acids) and of organic acids (such as, acetic acid, benzenesulfonic, benzoic, methanesulfonic, and p-toluenesulfonic acids) . Compounds of the present teachings with acidic groups such as carboxylic acids can form pharmaceutically acceptable salts with pharmaceutically acceptable base (s) . Suitable pharmaceutically acceptable basic salts include e.g., ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts) . Compounds with a quaternary ammonium group also contain a counteranion such as chloride, bromide, iodide, acetate, perchlorate and the like. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates and salts with amino acids such as glutamic acid.

The term “pharmaceutically acceptable carrier” refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.

As used herein, the term "subject" refers to human and non-human animals, including veterinary subjects. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, mice, rabbits, sheep, dog, cat, horse, cow, chickens, amphibians, and reptiles. In a preferred embodiment, the subject is a human and may be referred to as a patient.

As used herein, the terms "treat, " "treating" or "treatment" refer, preferably, to an action to obtain a beneficial or desired clinical result including, but not limited to, alleviation or amelioration of one or more signs or symptoms of a disease or condition, diminishing the extent of disease, stability (i.e., not worsening) of the state of disease, amelioration or palliation of the disease state, diminishing rate of or time to progression, and remission (whether partial or total) . "Treatment" can also mean prolonging survival as compared to expected survival in the absence of treatment. Treatment does not need to be curative.

A "therapeutically effective amount" is that amount sufficient to treat a disease in a subject. A therapeutically effective amount can be administered in one or more administrations. In one aspect, a therapeutically effective amount refers to a dosage of from about 0.01 to about 100 mg/kg body weight/day.

The terms "administer, " "administering" or "administration" include any method of delivery of a pharmaceutical composition or agent into a subject's system or to a particular region in or on a subject. In certain embodiments of the invention, an agent is administered intravenously, intramuscularly, subcutaneously, intradermally, intranasally, orally, transcutaneously, or mucosally. In a preferred embodiment, an agent is administered intravenously. In another preferred embodiment, an agent is administered orally. Administering an agent can be performed by a number of people working in concert. Administering an agent includes, for example, prescribing an agent to be administered to a subject and/or providing instructions, directly or through another, to take a specific agent, either by self-delivery, e.g., as by oral delivery, subcutaneous delivery, intravenous delivery through a central line, etc.; or for delivery by a trained professional, e.g., intravenous delivery, intramuscular delivery, intratumoral delivery, etc.

3. Compounds

In a first embodiment, the compound of Formula I is of the Formula II or III:

or a pharmaceutically acceptable salt thereof, wherein R^Q, R^T, X, L and A are as described above for Formula I.

In a second embodiment, the compound of Formula I is of the Formula III:

In a third embodiment, X in the compound of Formula I, II, or III is absent or -NR^W, wherein R^W is H or CH₃, wherein the remaining variables are as described above for Formula I. Alternatively, as part of a third embodiment, X in the compound of Formula I, II, or III is absent or -NH, wherein the remaining variables are as described above for Formula I.

In a fourth embodiment, R^Q and R^T in the compound of Formula I, II, or III are each independently selected from H and (C₁-C₄) alkyl, wherein the remaining variables are as described above for Formula I or the third embodiment. Alternatively, as part of a fourth embodiment, R^Q and R^T in the compound of Formula I, II, or III are each independently selected from H, CH (CH₃) ₂, and CH₃, wherein the remaining variables are as described above for Formula I or the third embodiment. In another alternative, as part of a fourth embodiment, R^Q in the compound of Formula I, II, or III is H and R^T is selected from H, CH (CH₃) ₂, and CH₃, wherein the remaining variables are as described above for Formula I or the third embodiment. In another alternative, as part of a fourth embodiment, R^Q and R^T are each H in the compound of Formula I, II, or III, wherein the remaining variables are as described above for Formula I or the third embodiment.

In a fifth embodiment, A in the compound of Formula I, II, or III is selected from

Z and Z’ are each independently selected from N or CH;

R¹, R^1A, and R^1AA are each independently selected from (C₁-C₄) alkyl, halo (C₁-C₄) alkyl, halo, (C₁-C₄) alkoxy, and halo (C₁-C₄) alkoxy;

R³, R^3A, and R^3AA are each independently selected from hydrogen, halo, (C₁-C₄) alkyl, halo (C₁-C₄) alkyl, (C₁-C₄) alkoxy, halo (C₁-C₄) alkoxy, - (C₀-C₄) alkylheterocyclyl, and - (C₀-C₄) heteroaryl, wherein said heterocyclyl and heteroaryl are each optionally substituted with 1 to 3 groups selected from R^Z;

R^Z is is selected from (C₁-C₄) alkyl, halo (C₁-C₄) alkyl, halo, (C₁-C₄) alkoxy, and halo (C₁-C₄) alkoxy;

R^a and R^b are each independently selected from hydrogen, (C₁-C₄) alkyl, and halo (C₁-C₄) alkyl;

R⁴ is (C₁-C₄) alkyl;

R⁵ and R^5A are each independently (C₁-C₄) alkyl;

R⁶ and R^6A are each independently hydrogen or (C₁-C₄) alkyl;

R⁷ is halo; and

the wavy line indicates the point of attachment to linker L, wherein the remaining variables are as described above for Formula I or any one of the third or fourth embodiment. Alternatively, as part of a fifth embodiment, A in the compound of Formula I, II, or III is selected from wherein the remaining variables are as described above for Formula I or any one of the third or fourth embodiment.

In a sixth embodiment, Z and Z’ are each CH, wherein the remaining variables are as described above for Formula I or any one of the third to fifth embodiments.

In a seventh embodiment, R¹, R^1A, and R^1AA are each (C₁-C₄) alkyl, wherein the remaining variables are as described above for Formula I or any one of the third to sixth embodiments. Alternatively, as part of a seventh embodiment, R¹, R^1A, and R^1AA are each isopropyl, wherein the remaining variables are as described above for Formula I or any one of the third to sixth embodiments.

In an eighth embodiment, R³, R^3A, and R^3AA are each independently selected from hydrogen, - (C₁-C₄) alkylheterocyclyl, and heteroaryl, wherein said heterocyclyl and heteroaryl are each optionally substituted with 1 to 3 groups selected from R^Z, wherein the remaining variables are as described above for Formula I or any one of the third to seventh embodiments. Alternatively, as part of an eighth embodiment, R^3A is hydrogen, wherein the remaining variables are as described above for Formula I or any one of the third to seventh embodiments. In another alternative, as part of an eighth embodiment, R^3AA is hydrogen, wherein the remaining variables are as described above for Formula I or any one of the third to seventh embodiments. In another alternative, as part of an eighth embodiment, R³ is selected from - (C₁-C₄) alkyl (5-to 6-membered) heterocyclyl and fused 8-10 membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R^Z, wherein the remaining variables are as described above for Formula I or any one of the third to seventh embodiments. In another alternative, as part of an eighth embodiment, R³ is selected from - (C₁-C₄) morpholinyl, indolyl, piperazinyl, wherein said morpholinyl, indolyl, and piperazinyl are each optionally substituted with 1 to 3 groups selected from R^Z, wherein the remaining variables are as described above for Formula I or any one of the third to seventh embodiments.

In a ninth embodiment, R^Z is (C₁-C₄) alkyl, wherein the remaining variables are as described above for Formula I or any one of the third to eighth embodiments.

In a tenth embodiment, R^a is hydrogen; and R^b is halo (C₁-C₄) alkyl, wherein the remaining variables are as described above for Formula I or any one of the third to ninth embodiments. Alternatively, as part of a tenth embodiment, R^a is hydrogen; and R^b is CH₂CF₃, wherein the remaining variables are as described above for Formula I or any one of the third to ninth embodiments.

In an eleventh embodiment, R⁴ is CH₃, wherein the remaining variables are as described above for Formula I or any one of the third to tenth embodiments.

In a twelfth embodiment, R⁵ and R^5A are each CH₃, wherein the remaining variables are as described above for Formula I or any one of the third to eleventh embodiments.

In a thirteenth embodiment, R⁶ and R^6A are each hydrogen, wherein the remaining variables are as described above for Formula I or any one of the third to twelfth embodiments.

In a fourteenth embodiment, R⁷ is chloro, wherein the remaining variables are as described above for Formula I or any one of the third to thirteenth embodiments.

In a fifteenth embodiment, L in the compound of Formula I, II, or III is selected from - (CH₂) _r-HET¹-X¹-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹- (CH₂) _q-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET²- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-HET²- (CH₂) _s-C (O) - (CH₂) _m-X²-, - (CH₂) _yOC (O) -, - (CH₂) _r-HET¹- (CH₂) _w-S-S-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET²- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET³-OC (O) -, - (CH₂) _r-HET²-X¹-HET³-OC (O) -, - (CH₂) _r-HET³-X¹- (CH₂) _s-OC (O) -, - (CH₂) _r-HET³-X¹-HET²-OC (O) -, -X¹-HET³-OC (O) -, -X¹-HET³-X²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹- (CH₂) _q-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹- (CH₂) _w-S-S-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-HET²- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _q-HET¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-HET¹- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²- (CH₂) _w-S-S-HET¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-C (O) - (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _q-HET¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-X¹- (CH₂) _q-HET³- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET³- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-HET³- (CH₂) _s-C (O) - (CH₂) _m-X²-, - (CH₂) _r-HET¹- (CH₂) _w-S-S-HET³- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET³- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET³-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET³- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET³-X¹-HET¹- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET³- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, -C (O) NH-CRR’ -C (O) NH-CRR’ -C (O) NHCRR’ C (O) NH (CH₂) _m-X²-, -C (O) NH-CRR’ -C (O) NH-CRR’ -C (O) NH-HET²- (CH₂) _s-OC (O) -, C (O) , -C (O) -X¹-HET¹-X²-, -X¹-HET¹-X², -C (O) -HET¹-X¹-HET²-X², and C (O) -X¹-O-;

m, q, r, s, v, w, and y are each independently 0 to 4;

R^f is selected from hydrogen and (C₁-C₄) alkyl;

X¹ and X² are each independently selected from CO and –CH₂-;

HET¹, HET², and HET³ are each independently selected from monocyclic heterocyclyl, bicyclic heterocyclyl, monocyclic cycloalkyl, and phenyl; and

R’ and R” are each independently selected from hydrogen and (C₁-C₄) alkyl, wherein said (C₁-C₄) alkyl is optionally substituted with phenyl or NHC (O) NH₂, wherein the remaining variables are as described above for Formula I or any one of the third to fourteenth embodiments. Alternatively, as part of a fifteenth embodiment, L is selected from C (O) , -C (O) -X¹-HET¹-X²-, -X¹-HET¹-X², -C (O) -HET¹-X¹-HET²-X², and C (O) -X¹-O-, wherein the remaining variables are as described above for Formula I or any one of the third to fourteenth embodiments.

In a sixteenth embodiment, X¹ and X² are each –CH₂-, wherein the remaining variables are as described above for Formula I or any one of the third to fifteenth embodiments.

In a seventeenth embodiment, HET¹, HET², and HET³ are each independently selected from monocyclic heterocyclyl and monocyclic cycloalkyl, wherein the remaining variables are as described above for Formula I or any one of the third to sixteenth embodiments. Alternatively, as part of a seventeenth embodiment, HET¹ and HET², and HET³ are each independently selected from piperizinyl and cyclohexyl, wherein the remaining variables are as described above for Formula I or any one of the third to sixteenth embodiments. In another alternative, as part of a seventeenth embodiment, L is selected from C (O) , and C (O) CH₂O*, wherein *indicates the point of attachment to A, wherein the remaining variables are as described above for Formula I or any one of the third to sixteenth embodiments.

Compounds having the Formula I, II, and III are further disclosed in the Exemplification and are included in the present disclosure. Pharmaceutically acceptable salts thereof as well as the neutral forms are included.

4. Uses, Formulation and Administration

Compounds and compositions described herein are generally useful as anticancer therapies. The terms "cancer" or "tumor" are well known in the art and refer to the presence, e.g., in a subject, of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, decreased cell death/apoptosis, and certain characteristic morphological features. Cancer cells are often in the form of a solid tumor. However, cancer also includes non-solid tumors, e.g., blood tumors, e.g., leukemia, wherein the cancer cells are derived from bone marrow. As used herein, the term "cancer" includes pre-malignant as well as malignant cancers. Cancers include, but are not limited to, acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia (monocytic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic) , acute T-cell leukemia, basal cell carcinoma, bile duct carcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myelocytic (granulocytic) leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, dysproliferative changes (dysplasias and metaplasias) , embryonal carcinoma, endometrial cancer, endotheliosarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen-receptor positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, heavy chain disease, hemangioblastoma, hepatoma, hepatocellular cancer, hormone insensitive prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphagioendotheliosarcoma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin and non-Hodgkin) , malignancies and hyperproliferative disorders of the bladder, breast, colon, lung, ovaries, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myelogenous leukemia, myeloma, myxosarcoma, neuroblastoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinomas, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung carcinoma, solid tumors (carcinomas and sarcomas) , small cell lung cancer, stomach cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular tumors, uterine cancer, and Wilms'tumor. Other cancers include primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gall bladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelium cancer, female genital tract cancer, uterine cancer, gestational trophoblastic disease, male genital tract cancer, seminal vesicle cancer, testicular cancer, germ cell tumors, endocrine gland tumors, thyroid cancer, adrenal cancer, pituitary gland cancer, hemangioma, sarcoma arising from bone and soft tissues, Kaposi's sarcoma, nerve cancer, ocular cancer, meningial cancer, glioblastomas, neuromas, neuroblastomas, Schwannomas, solid tumors arising from hematopoietic malignancies such as leukemias, metastatic melanoma, recurrent or persistent ovarian epithelial cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumors, colorectal cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small-cell lung cancer, malignant glioma, epithelial ovarian cancer, primary peritoneal serous cancer, metastatic liver cancer, neuroendocrine carcinoma, refractory malignancy, triple negative breast cancer, HER2-amplified breast cancer, nasopharageal cancer, oral cancer, biliary tract, hepatocellular carcinoma, squamous cell carcinomas of the head and neck (SCCHN) , non-medullary thyroid carcinoma, recurrent glioblastoma multiforme, neurofibromatosis type 1, CNS cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral/lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumor, triple negative breast cancer, colorectal cancer, sarcoma, melanoma, renal carcinoma, endometrial cancer, thyroid cancer, rhabdomysarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumor, mantle cell lymphoma, and refractory malignancy.

"Solid tumor, " as used herein, is understood as any pathogenic tumor that can be palpated or detected using imaging methods as an abnormal growth having three dimensions. A solid tumor is differentiated from a blood tumor such as leukemia. However, cells of a blood tumor are derived from bone marrow; therefore, the tissue producing the cancer cells is a solid tissue that can be hypoxic.

"Tumor tissue” or “tumorous tissue" are understood as cells, extracellular matrix, and other naturally occurring components associated with the solid tumor.

A specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound described herein in the composition will also depend upon the particular compound in the composition.

EXEMPLIFICATION

Chemical Synthesis

The representative examples that follow are intended to help illustrate the present disclosure, and are not intended to, nor should they be construed to, limit the scope of the invention. General starting materials used were obtained from commercial sources or prepared in other examples, unless otherwise noted.

Preparation of Compounds

The compounds claimed herein were prepared following the procedures outlined in the following protocols.

Example 1: Compound 001 synthesis

A representative synthesis scheme for compound 001 is shown in below. Specific synthesis routes of intermediates are also shown.

Intermediate 2: (9H-fluoren-9-yl) methyl ( (1R, 9S) -9-ethyl-5-fluoro-9-hydroxy-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-1-yl) carbamate

To a solution of compound 1 (0.80 g, 1.51 mmol, 1.0 eq) and 0.75M NaHCO₃ (20.06 mL, 0.75 mol/L, 10.0 eq) in MeCN (40 mL) was added Fmoc-Osu (0.75 g, 2.26 mmol, 1.5 eq) at 25 ℃. The reaction mixture was stirred at 25 ℃ for 18 h. The reaction mixture was concentrated and purified by FC (DCM: MeOH=1: 20) to give the compound 2 (0.95 g, 96%yield) as yellow soild.

Intermediate 4: (1R, 9S) -1- ( ( ( (9H-fluoren-9-yl) methoxy) carbonyl) amino) -9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12Hbenzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (tert-butoxycarbonyl) glycinate

To a stirred solution of compound 3 (0.48 g, 2.73 mmol, 2.0 eq) in DCM (40 mL) at 0 ℃ was added EDCI (0.53 g, 2.73 mmol, 2.0 eq) and DMAP (0.05 g, 0.41 mmol, 0.3 eq) at 0 ℃ for 4 min. Then compound 2 (0.91 g, 1.37 mmol, 1.0 eq) was added to the solution and stirred at 25℃ for 18 h. The reaction mixture was concentrated and purified by FC (DCM: MeOH=1: 20) to give the compound 4 (0.85 g, 72%yield) as yellow soild.

Intermediate 5: (1R, 9S) -1- ( ( ( (9H-fluoren-9-yl) methoxy) carbonyl) amino) -9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl glycinate

To a solution of compound 4 (800 mg, 0.982 mmol, 1.0 eq) in DCM (11.25 mL) was added TFA (2.25 ml) at 25 ℃. The mixture was stirred at 25℃ for 1 h. The mixture was concentrated under reduced pressure to afford the desired product (850 mg, crude) as yellow solid.

Intermediate 7: (1R, 9S) -1- ( ( ( (9H-fluoren-9-yl) methoxy) carbonyl) amino) -9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- (morpholino methyl) phenyl) isoxazole-3-carbonyl) glycinate

To a mixture of compound 6 (237 mg, 0.54 mmol, 1.1 eq) , PyBop (383 mg, 0.736 mmol, 1.5 eq) and DIPEA (0.49 mL, 2.95 mmol, 6.0 eq) in DCM (20 mL) was added compound 5 (351 mg, 0.49 mmol, 1.0 eq) at 25 ℃. The mixture was stirred at 25℃ for 18 h. The mixture was concentrated and purified by Prep-HPLC (TFA) to afford the compound 7 (0.27 g, 48%yield) as yellow soild.

Compound 001: (1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- (morpholinomethyl) phenyl) isoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate

To a solution of compound 4 (270 mg, 0.238 mmol, 1.0 eq) in DMF (12.92 mL) was added Piperdine (0.32 ml) at 0 ℃. The mixture was stirred at 0 ℃ for 10 min. The mixture was concentrated and purified by Prep-HPLC (TFA) to afford the desired product Compound 001 (210.2 mg, TFA salt, 86%yield) as yellow solid. ¹H NMR (400 MHz, DMSO) δ10.07-10.04 (m, 1H) , 9.83 (s, 1H) , 9.70 (s, 1H) , 9.38-9.36 (m, 1H) , 8.54-8.50 (m, 3H) , 7.82-7.80 (m, 1H) , 7.38-7.36 (m, 4H) , 7.23-7.21 (m, 1H) , 6.75 (s, 1H) , 6.42 (s, 1H) , 5.73-5.71 (m, 1H) , 5.57-5.53 (m, 2H) , 5.46-5.44 (m, 1H) , 5.14-5.12 (m, 1H) , 4.29-4.25 (m, 4H) , 3.94-3.92 (m, 2H) , 3.66-3.62 (m, 3H) , 3.17-3.07 (m, 3H) , 2.99-2.95 (m, 2H) , 2.46-2.40 (m, 3H) , 2.19-2.11 (m, 4H) , 0.92-0.89 (m, 9H) . LCMS: m/z (ES+) (M+H) ⁺ = 913.2, Rt = 0.879 min.

The following additional compounds were made according to the general procedure above using the appropriate starting materials.

Compound 001

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- (morpholinomethyl) phenyl) isoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ10.07-10.04 (m, 1H) , 9.83 (s, 1H) , 9.70 (s, 1H) , 9.38-9.36 (m, 1H) , 8.54-8.50 (m, 3H) , 7.82-7.80 (m, 1H) , 7.38-7.36 (m, 4H) , 7.23-7.21 (m, 1H) , 6.75 (s, 1H) , 6.42 (s, 1H) , 5.73-5.71 (m, 1H) , 5.57-5.53 (m, 2H) , 5.46-5.44 (m, 1H) , 5.14-5.12 (m, 1H) , 4.29-4.25 (m, 4H) , 3.94-3.92 (m, 2H) , 3.66-3.62 (m, 3H) , 3.17-3.07 (m, 3H) , 2.99-2.95 (m, 2H) , 2.46-2.40 (m, 3H) , 2.19-2.11 (m, 4H) , 0.92-0.89 (m, 9H) . LCMS (ESI) : R_T = 0.879 min, m/z found 913.2 [M+H] ⁺.

Compound 002

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- (morpholinomethyl) phenyl) isoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ9.96 (s, 1H) , 9.82 (s, 1H) , 9.68 (s, 1H) , 9.39-9.37 (m, 1H) , 8.51-8.48 (m, 3H) , 7.83-7.80 (m, 1H) , 7.38-7.34 (m, 4H) , 7.22-7.18 (m, 1H) , 6.75 (s, 1H) , 6.42 (s, 1H) , 5.74-5.68 (m, 1H) , 5.56-5.52 (m, 2H) , 5.46-5.42 (m, 1H) , 5.13-5.11 (m, 1H) , 4.28-4.22 (m, 4H) , 3.94-3.92 (m, 2H) , 3.60-3.56 (m, 2H) , 3.24-2.91 (m, 6H) , 2.54-2.52 (m, 1H) , 2.44-2.40 (m, 3H) , 2.23-2.10 (m, 3H) , 0.94-0.90 (m, 9H) . LCMS (ESI) : R_T = 0.844 min, m/z found 913.3 [M+H] ⁺.

Compound 003

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ10.15 (s, 1H) , 9.75 (s, 1H) , 9.63 (t, J = 6.4 Hz, 1H) , 8.60-8.52 (m, 3H) , 7.84 (d, J = 10.8 Hz, 1H) , 7.48-7.39 (m, 4H) , 7.17 (s, 1H) , 6.67 (s, 1H) , 6.30 (s, 1H) , 5.74-5.65 (m, 1H) , 5.53-5.41 (m, 3H) , 5.12 (s, 1H) , 4.30-4.24 (m, 1H) , 4.14-4.08 (m, 2H) , 3.99-3.91 (m, 3H) , 3.38-3.28 (m, 6H) , 3.17-3.08 (m, 4H) , 2.96-2.86 (m, 4H) , 2.43 (s, 3H) , 2.22-2.10 (m, 4H) , 0.91 (t, J = 7.6 Hz, 3H) , 0.85 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : R_T = 0.829 min, m/z found 1051.4 [M+H] ⁺.

Compound 004

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ10.12 (s, 1H) , 9.73 (s, 1H) , 9.63 (t, J = 6.4 Hz, 1H) , 8.47 (s, 3H) , 7.85 (d, J = 10.8 Hz, 1H) , 7.48-7.39 (m, 4H) , 7.17 (s, 1H) , 6.66 (s, 1H) , 6.29 (s, 1H) , 5.71 (d, J = 19.2 Hz, 1H) , 5.58-5.42 (m, 3H) , 5.10 (s, 1H) , 4.29-3.94 (m, 6H) , 3.29-2.90 (m, 14H) , 2.43 (s, 4H) , 2.20-2.15 (m, 3H) , 0.92 (t, J = 7.2 Hz, 3H) , 0.84 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 0.797 min, m/z found 1051.4 [M+H] +.

Compound 005

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl 2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.87 (s, 1H) , 9.76 (s, 1H) , 9.65 (t, J = 6.4 Hz, 1H) , 8.55 (s, 3H) , 7.84 (d, J = 10.8 Hz, 1H) , 7.54-7.52 (m, 2H) , 7.45-7.43 (m, 2H) , 7.11 (s, 1H) , 6.70 (s, 1H) , 6.31 (s, 1H) , 5.73 (d, J = 19.2 Hz, 1H) , 5.55 (s, 2H) , 5.45 (d, J = 19.2 Hz, 1H) , 5.13 (s, 1H) , 4.36 (s, 2H) , 3.98-3.94 (m, 3H) , 3.40–3.01 (m, 9H) , 2.95-2.91 (m, 2H) , 2.72-2.68 (m, 2H) , 2.43 (s, 3H) , 2.22–2.12 (m, 3H) , 0.94 (t, J = 7.4 Hz, 3H) , 0.88 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 0.947 min, m/z found 821.2 [M+H] +.

Compound 006

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl-2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ10.11 (s, 1H) , 9.75 (s, 1H) , 9.63 (t, J = 6.4 Hz, 1H) , 8.55 (s, 3H) , 7.84 (d, J = 10.8 Hz, 1H) , 7.50 (d, J = 8.4z Hz, 2H) , 7.42 (d, J = 8.4 Hz, 2H) , 7.11 (s, 1H) , 6.65 (s, 1H) , 6.30 (s, 1H) , 5.72 (d, J = 19.2 Hz, 1H) , 5.59-5.43 (m, 3H) , 5.11 (s, 1H) , 4.33 (s, 2H) , 3.96-3.92 (m, 2H) , 3.77-3.65 (m, 3H) , 3.31-3.07 (m, 10H) , 2.82-79 (m, 1H) , 2.41 (s, 3H) , 2.21- 2.15 (m, 3H) , 0.93 (t, J = 7.6 Hz, 3H) , 0.80 (d, J = 7.2 Hz, 6H) . LCMS (ESI) : RT = 1.565 min, m/z found 821.2 [M+H] +.

Compound 007

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl-2- (4- ( (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindolin-5-yl) methyl) piperazin-1-yl) acetate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ10.02 (s, 1H) , 9.69-9.65 (m, 2H) , 8.54 (s, 3H) , 7.83 (d, J = 10.8 Hz, 1H) , 7.41-7.38 (m, 3H) , 7.09 (s, 1H) , 7.03 (s, 1H) , 6.41 (s, 1H) , 5.75-5.73 (m, 1H) , 5.54 (s, 2H) , 5.46-5.42 (m, 1H) , 5.13 (s, 1H) , 4.83-4.79 (m, 4H) , 4.32 (s, 2H) , 3.77-3.59 (m, 3H) , 3.34-2.95 (m, 10H) , 2.45-2.41 (m, 3H) , 2.21-2.15 (m, 3H) , 1.14 (d, J = 6.8 Hz, 6H) , 0.92 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT = 1.255 min, m/z found 871.7 [M+H] +.

Compound 008

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl-2- (4- ( (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindolin-5-yl) methyl) piperazin-1-yl) acetate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ10.04 (s, 1H) , 9.68 (s, 1H) , 8.53 (s, 3H) , 7.82 (d, J = 10.8 Hz, 1H) , 7.48-7.34 (m, 3H) , 7.08 (s, 1H) , 7.01 (s, 1H) , 6.39 (s, 1H) , 5.73-5.67 (m, 1H) , 5.57-5.4 (m , 3H) , 5.09 (s, 1H) , 4.89-4.64 (m, 4H) , 4.27 (s, 2H) , 3.72-3.53 (m, 3H) , 3.26-2.97 (m, 11H) , 2.40 (s, 3H) , 2.20-2.12 (m , 3H) , 1.11 (d, J = 6.8 Hz, 6H) , 0.91 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT = 1.109 min, m/z found 871.7 [M+H] +.

Compound 009

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindoline-5-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ10.15 –10.00 (m, 1H) , 9.63 (s, 1H) , 9.10 (s, 1H) , 8.43 (s, 3H) , 7.86 –7.80 (m, 3H) , 7.46 –7.38 (m, 1H) , 7.26 (s, 1H) , 7.03 (s, 1H) , 6.39 (s, 1H) , 5.71 (d, J = 19.2 Hz, 1H) , 5.53 (s, 2H) , 5.43 (d, J = 19.2 Hz, 1H) , 5.12 (s, 1H) , 4.82 –4.74 (m, 4H) , 4.38 –4.32 (m, 1H) , 4.18 –4.12 (m, 1H) , 3.12 –3.05 (m, 3H) , 2.34 –2.13 (m, 4H) , 1.13 (d, J = 6.8 Hz, 6H) , 0.91 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT = 1.238 min, m/z found 816.2 [M+H] +.

Compound 010

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindoline-5-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ10.09 (s, 1H) , 9.72 (s, 1H) , 9.08 (s, 1H) , 8.48 (s, 3H) , 7.82-7.77 (m, 3H) , 7.43-7.27 (m, 2H) , 7.02 (s, 1H) , 6.38 (s, 1H) , 5.72-5.65 (m, 1H) , 5.51 (s, 2H) , 5.07 (s, 1H) , 4.87-4.73 (m, 4H) , 4.34-4.28 (m, 1H) , 4.17-4.13 (m, 1H) , 3.09-3.05 (m, 2H) , 2.42 (s, 3H) , 2.21-2.14 (m, 3H) , 2.02-1.96 (m, 1H) , 1.22 (s, 2H) , 1.11 (d, J = 6.8 Hz, 6H) , 0.91 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT = 0.998 min, m/z found 816.2 [M+H] +.

Compound 011

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2, 4-dihydroxy-5-isopropylphenyl) -4-phenylisoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.76 (s, 1H) , 9.64 (s, 1H) , 9.40 (t, J = 6.0 Hz, 1H) , 8.46-8.42 (m, 3H) , 7.78 (d, J = 10.8 Hz, 1H) , 7.29-2.25 (m, 2H) , 7.23-7.17 (m, 4H) , 6.76 (s, 1H) , 6.42 (s, 1H) , 5.72 (d, J = 19.2 Hz, 1H) , 5.54 (s, 2H) , 5.43 (d, J = 19.2 Hz, 1H) , 5.15-5.11 (m, 1H) , 4.34-4.28 (m, 1H) , 4.18-4.12 (m, 1H) , 3.33-3.29 (m, 1H) , 3.15-3.11 (m, 1H) , 2.99-2.95 (m, 1H) , 2.55-2.51 (m, 1H) , 2.43 (s, 3H) , 2.21-2.17 (m, 1H) , 2.13-2.09 (m, 2H) , 0.93-0.87 (m, 9H) . LCMS (ESI) : RT = 1.129 min, m/z found 813.9 [M+H] +.

Compound 012

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2, 4-dihydroxy-5-isopropylphenyl) -4-phenylisoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.76 (s, 1H) , 9.63 (s, 1H) , 9.40 (t, J = 6.0 Hz, 1H) , 8.45 (s, 3H) , 7.76 (d, J = 10.8 Hz, 1H) , 7.25-7.22 (m, 3H) , 7.17-7.12 (m, 3H) , 6.75 (s, 1H) , 6.41 (s, 1H) , 5.71 (d, J = 19.2 Hz, 1H) , 5.55 (s, 2H) , 5.47-5.43 (m, 1H) , 5.12 (s, 1H) , 4.33-4.27 (m, 1H) , 4.19-4.13 (m, 1H) , 3.27-3.10 (m, 2H) , 2.99-2.95 (m, 1H) , 2.43 (s, 3H) , 2.24-2.19 (m, 1H) , 2.15-2.11 (m, 2H) , 0.93-0.89 (m, 9H) . LCMS (ESI) : RT = 1.419 min, m/z found 814.2 [M+H] +.

Compound 013

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.76 (s, 1H) , 9.62 (m, 1H) , 9.10 (s, 1H) , 8.50 (s, 3H) , 8.35-8.31 (m, 1H) , 7.85-7.82 (m, 1H) , 7.40-7.37 (m, 4H) , 7.16 (s, 1H) , 6.65 (s, 1H) , 6.33 (s, 1H) , 5.74-5.70 (m, 1H) , 5.53 (s, 2H) , 5.46-5.42 (m, 1H) , 5.12 (s, 1H) , 4.16-4.12 (m, 5.8 Hz, 1H) , 4.02-3.98 (m, 1H) , 3.96-3.91 (m, 2H) , 3.38-3.29 (m, 6H) , 3.10-3.02 (m, 2H) , 2.96–2.88 (m, 4H) , 2.76-2.74 (m, 1H) , 2.52-2.48 (m, 3H) , 2.43 (s, 3H) , 2.23-2.11 (m, 4H) , 1.82-1.80 (m, 5H) , 1.47-1.35 (m, 2H) , 0.93-0.90 (m, 5H) , 0.84 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 0.996 min, m/z found 1133.9 [M+H] +.

Compound 014

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.93 (s, 1H) , 9.62-9.60 (m, 1H) , 8.58 (s, 3H) , 8.32-8.30 (m, 1H) , 7.84-7.82 (m, 1H) , 7.44-7.40 (m, 4H) , 7.19-7.16 (m, 1H) , 6.66 (s, 1H) , 6.33 (s, 1H) , 5.72-5.70 (m, 1H) , 5.56-5.54 (m, 2H) , 5.45-5.43 (m, 1H) , 5.11 (s, 1H) , 4.15-3.97 (m, 4H) , 3.32-3.28 (m, 2H) , 3.21-3.03 (m, 4H) , 2.97-2.79 (m, 6H) , 2.77-2.75 (m, 1H) , 2.57-2.51 (m, 3H) , 2.42 (s, 3H) , 2.22-2.07 (m, 4H) , 1.84-1.64 (m, 5H) , 1.44-1.32 (m, 2H) , 0.94-0.90 (m, 5H) , 0.85 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : R_T = 0.952 min, m/z found 1133.9 [M+H] ⁺

Compound 015

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (1-methyl-1H-indol-6-yl) isoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.70 (s, 1H) , 9.56 (s, 1H) , 9.38-9.35 (m, 1H) , 8.39-8.36 (m, 3H) , 7.77 (d, J = 10.8 Hz, 1H) , 7.37-7.32 (m, 2H) , 7.25 (d, J = 3.2 Hz, 1H) , 7.20 (s, 1H) , 6.88-6.82 (m, 1H) , 6.81 (s, 1H) , 6.39 (s, 1H) , 6.31 (d, J = 3.2 Hz, 1H) , 5.69 (d, J = 19.2 Hz, 1H) , 5.53 (s, 2H) , 5.41 (d, J = 19.2 Hz, 1H) , 5.12-5.07 (m, 1H) , 4.38-4.32 (m, 1H) , 4.21-4.15 (m, 1H) , 3.59 (s, 3H) , 2.97-2.93 (m, 2H) , 2.42 (s, 3H) , 2.18-2.08 (m, 4H) , 1.66-1.62 (m, 1H) , 0.89-0.84 (m, 9H) . LCMS (ESI) : RT =0.947 min, m/z found 821.2 [M+H] +.

Compound 016

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (2- ( (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindolin-5-yl) oxy) acetyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ10.03 (s, 1H) , 9.60 (s, 1H) , 8.67 –8.60 (m, 1H) , 8.48 –8.30 (m, 2H) , 7.81 (d, J = 10.8 Hz, 1H) , 7.21 –7.14 (m, 2H) , 7.01 (s, 1H) , 6.95 –6.85 (m, 2H) , 6.38 (s, 1H) , 5.70 (d, J = 19.2 Hz, 1H) , 5.52 (s, 2H) , 5.44 (d, J = 19.2 Hz, 1H) , 5.08 (s, 1H) , 4.65 (s, 4H) , 4.51 (s, 2H) , 4.25 –4.19 (m, 1H) , 4.09 –4.03 (m, 1H) , 3.11 –3.05 (m, 2H) , 2.42 –2.37 (m, 5H) , 2.20 –2.10 (m, 4H) , 1.12 (d, J = 6.8 Hz, 6H) , 0.90 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT = 1.354 min, m/z found 846.4 [M+H] +.

Compound 017

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzoyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.99 (s, 1H) , 9.71 (s, 1H) , 9.64 (t, J = 6.4 Hz, 1H) , 9.22 (t, J = 5.6 Hz, 1H) , 8.45 (s, 3H) , 7.90 (d, J = 8.8 Hz, 2H) , 7.85 (d, J = 10.8 Hz, 1H) , 7.42 (d, J = 8.8 Hz, 2H) , 7.26 (s, 1H) , 6.74 (s, 1H) , 6.27 (s, 1H) , 5.71 (d, J = 19.6 Hz, 1H) , 5.53 (s, 2H) , 5.43 (d, J = 19.6 Hz, 1H) , 5.11 (s, 1H) , 4.36-4.18 (m, 1H) , 4.17-4.14 (m, 1H) , 3.96-3.92 (m, 2H) , 3.30 (s, 2H) , 3.16-3.11 (m, 1H) , 2.94-2.91 (m, 1H) , 2.43 (s, 3H) , 2.19-2.14 (m, 3H) , 0.92-0.88 (m, 9H) . LCMS (ESI) : RT = 1.380 min, m/z found 939.2 [M+H] +.

Compound 018

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2-amino-4-chloro-7- ( (4-methoxy-3, 5-dimethylpyridin-2-yl) methyl) -7H-pyrrolo [2, 3-d] pyrimidin-5-yl) pent-4-ynoyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ8.56 (t, J = 6.0 Hz, 1H) , 8.42 –8.39 (m, 3H) , 8.04 (s, 1H) , 7.86 (d, J = 10.8 Hz, 1H) , 7.21 (s, 1H) , 7.18 (s, 1H) , 6.67 (s, 2H) , 5.76 –5.68 (m, 1H) , 5.51 (s, 2H) , 5.47 –5.40 (m, 1H) , 5.25 (s, 2H) , 5.14 –5.11 (m, 1H) , 4.21 –4.15 (m, 1H) , 4.00 –3.94 (m, 1H) , 3.74 –3.71 (m, 4H) , 2.45 –2.39 (m, 6H) , 2.25 –2.21 (m, 6H) , 2.19 –2.14 (m, 6H) , 0.88 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT=0.649 min, m/z found 444.6 [1/2M+H] ⁺.

Compound 019

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- (morpholinomethyl) phenyl) isoxazole-3-carbonyl) -L-valinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ 9.83 (s, 1H) , 9.69 (s, 1H) , 9.34 (s, 1H) , 8.43 (s, 3H) , 7.65 (d, J = 10.8 Hz, 1H) , 7.38-7.36 (m, 3H) , 7.18 (s, 1H) , 6.81 (s, 1H) , 6.46 (s, 1H) , 5.71 (d, J = 19.2 Hz, 1H) , 5.54 (s, 2H) , 5.45 (d, J = 19.2 Hz, 1H) , 5.10 (s, 1H) , 4.43-4.39 (m, 4H) , 3.96-3.92 (m, 4H) , 3.14-3.10 (m, 4H) , 3.09-3.01 (m, 3H) , 2.21-2.17 (m, 5H) , 1.67-1.63 (m, 1H) , 1.21-1.19 (m, 2H) , 0.95-0.91 (m, 15H) . LCMS (ESI) : R_T = 1.683 min, m/z found 956.4 [M+H] ⁺.

Compound 020

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- (morpholinomethyl) phenyl) isoxazole-3-carbonyl) -D-valinate 2, 2, 2-trifluoroacetate ¹HNMR (400 MHz, DMSO) δ 9.83 (s, 1H) , 9.69 (s, 1H) , 9.35 (d, J = 10.8 Hz, 1H) , 8.40 (s, 2H) , 7.68 (d, J = 10.8 Hz, 1H) , 7.44-7.40 (m, 4H) , 7.17 (s, 1H) , 6.82 (s, 1H) , 6.46 (s, 1H) , 5.71 (d, J = 19.2 Hz, 1H) , 5.54 (s, 2H) , 5.43 (d, J = 19.2 Hz, 1H) , 5.11 (s, 1H) , 4.44-4.38 (m, 2H) , 3.96 (s, 1H) , 3.59-3.51 (m, 5H) , 3.13-3.07 (m, 4H) , 3.05–2.95 (m, 3H) , 2.19-2.15 (m, 5H) , 1.66-1.64 (m, 1H) , 1.21-1.17 (m, 2H) , 0.97-0.91 (m, 15H) . LCMS (ESI) : R_T = 1.703 min, m/z found 956.4 [M+H] ⁺.

Compound 021

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- (5- (2, 4-dihydroxy-5-isopropylphenyl) -4- (4- ( (4-methylpiperazin-1-yl) methyl) phenyl) isoxazole-3-carbonyl) glycinate 2, 2, 2-trifluoroacetate ¹H NMR (400 MHz, DMSO) δ9.79 (s, 1H) , 9.68 (s, 1H) , 9.35 (t, J = 6 Hz, 1H) , 8.47 (s, 3H) , 7.80 (d, J = 10.8 Hz, 1H) , 7.24 (m, 5H) , 6.70 (s, 1H) , 6.43 (s, 1H) , 5.71 (d, J = 19.2 Hz, 1H) , 5.54 (s, 2H) , 5.43 (d, J = 19.2 Hz, 1H) , 5.12 (s, 1H) , 4.24 (m, 2H) , 3.53 (s, 2H) , 3.30 –3.25 (m, 2H) , 3.11 (s, 1H) , 2.96 (m, 4H) , 2.76 (s, 3H) , 2.68 –2.66 (m, 1H) , 2.42 (s, 3H) , 2.35 –2.30 (m, 2H) , 2.21 –2.07 (m, 3H) , 0.92 (d, J = 7.2 Hz, 3H) , 0.88 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 0.721 min, m/z found 463.8 [1/2M+H] +.

Compound 022

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- ( (1r, 4s) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-alaninate formate ¹H NMR (400 MHz, DMSO) δ10.51 (s, 1H) , 9.60 (s, 1H) , 8.38 (d, J = 6.8 Hz, 1H) , 7.61 (d, J = 10.8 Hz, 1H) , 7.36 (dd, J = 20.4, 8.4 Hz, 4H) , 7.06 (s, 1H) , 6.62 (s, 1H) , 6.36 (s, 1H) , 5.66 –5.39 (m, 4H) , 4.36 (m, 2H) , 4.02 –3.94 (m, 2H) , 3.50 (s, 2H) , 3.09 (s, 2H) , 2.90 –2.86 (m, 1H) , 2.69 (d, J = 1.8 Hz, 3H) , 2.38 (s, 5H) , 2.35 (s, 4H) , 2.13 (dd, J = 14.9, 7.3 Hz, 6H) , 1.79 (d, J = 37.3 Hz, 5H) , 1.52 (s, 3H) , 1.40 (d, J = 7.3 Hz, 3H) , 1.26 (s, 2H) , 0.91 (m, 5H) , 0.80 (m, 6H) . LCMS (ESI) : RT = 0.658 min, m/z found 574.4 [1/2M+H] +.

Compound 023

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- ( (1r, 4s) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-valinate formate ¹H NMR (400 MHz, DMSO) δ 10.50 (s, 1H) , 9.61 (t, J = 6.4 Hz, 1H) , 8.30 –8.21 (m, 1H) , 7.56 (d, J = 10.8 Hz, 1H) , 7.37 –7.31 (m, 4H) , 7.08 –7.05 (m, 1H) , 6.62 –6.60 (m, 1H) , 6.34 (s, 1H) , 5.66 –5.58 (m, 1H) , 5.53 –5.44 (m, 2H) , 5.42 –5.34 (m, 1H) , 4.39 –4.36 (m, 1H) , 4.14 –4.09 (m, 1H) , 4.00 –3.91 (m, 2H) , 3.51 –3.44 (m, 2H) , 3.07 –2.84 (m, 4H) , 2.67 (s, 3H) , 2.42 –2.33 (m, 13H) , 2.19 –2.04 (m, 8H) , 1.84 –1.70 (m, 4H) , 1.57 –1.34 (m, 4H) , 0.97 –0.88 (m, 9H) , 0.83 –0.76 (m, 6H) . LCMS (ESI) : RT = 0.670 min, m/z found 588.3 [1/2M+H] +.

Compound 024

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl-N- ( (1r, 4s) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -N-methylglycinate formate ¹H NMR (400 MHz, DMSO) (400 MHz, DMSO) δ 10.46 (s, 1H) , 9.58 (s, 1H) , 7.64 (t, J = 10.4 Hz, 1H) , 7.40 –7.26 (m, 4H) , 7.12 (d, J = 14.4 Hz, 1H) , 6.62 (d, J = 4.4 Hz, 1H) , 6.34 (d, J = 3.6 Hz, 1H) , 5.63 (d, J = 18.8 Hz, 1H) , 5.51 (d, J = 6.4 Hz, 2H) , 5.41 –5.31 (m, 1H) , 4.48 –4.13 (m, 2H) , 3.96 (s, 2H) , 3.09 (s, 2H) , 2.78 (s, 1H) , 2.08 -1.90 (m, 11H) , 1.83 –1.00 (m, 11H) , 0.91 (t, J = 6.4 Hz, 4H) , 0.80 (t, J = 9.2 Hz, 6H) . LCMS (ESI) : RT = 1.380 min, m/z found 574.2 [1/2M+H] +.

Compound 025

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl- ( (1r, 4s) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-alaninate formate ¹H NMR (400 MHz, DMSO) δ 10.50 (s, 1H) , 9.58 (s, 1H) , 8.36 (d, J = 6.8 Hz, 1H) , 7.59 (d, J = 10.8 Hz, 1H) , 7.34 (m, 4H) , 7.04 (s, 1H) , 6.60 (s, 1H) , 6.34 (s, 1H) , 5.70 –5.26 (m, 4H) , 4.32 (m, 2H) , 4.03 –3.85 (m, 2H) , 3.48 (s, 2H) , 3.06 (s, 2H) , 2.89 –2.83 (m, 1H) , 2.68 –2.66 (m, 3H) , 2.36 (s, 5H) , 2.34 –2.32 (m, 4H) , 2.16 –2.04 (m, 6H) , 1.78 (d, J = 40.8 Hz, 5H) , 1.50 (s, 3H) , 1.37 (d, J = 7.2 Hz, 3H) , 1.23 (s, 2H) , 0.89 (m, 5H) , 0.78 (d, J = 7.0 Hz, 6H) . LCMS (ESI) : RT = 0.647 min, m/z found 574.4 [1/2M+H] +.

Compound 026

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4s) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-valinate formate ¹H NMR (400 MHz, DMSO) δ 10.45 (s, 1H) , 9.52 (t, J = 6.4 Hz, 1H) , 8.22-8.20 (m, 1H) , 8.11-8.07 (m, 1H) , 7.51-7.47 (m, 1H) , 7.30-7.26 (m, 4H) , 7.03-6.97 (m, 1H) , 6.56-6.52 (m, 1H) , 6.28 (s, 1H) , 5.56-5.52 (m, 1H) , 5.42 (s, 2H) , 5.35-5.23 (m, 1H) , 4.35 (s, 1H) , 4.09-4.01 (m, 2H) , 3.91-3.79 (m, 4H) , 3.18-3.14 (m, 4H) , 2.98-2.90 (m, 3H) , 2.82-2.70 (m, 3H) , 2.29-2.24 (m, 5H) , 2.07-2.01 (m, 8H) , 1.83-1.62 (m, 5H) , 1.44-1.38 (m, 4H) , 1.21-1.15 (m, 2H) , 0.94-0.86 (m, 3H) , 0.84-0.76 (m, 6H) , 0.74-0.66 (m, 6H) . LCMS (ESI) : RT = 1.445 min, m/z found 1176.4 [M+H] +.

Compound 027

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-histidyl-L-valinate. LCMS (ESI) : RT = 5.088 min, m/z found 1312.6 [M+H] ⁺.

Compound 028

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-histidyl-D-valinate. LCMS (ESI) : RT = 5.176 min, m/z found 1312.6 [M+H] ⁺.

Compound 029

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-histidyl-L-valinate. ¹H NMR (400 MHz, DMSO) δ 11.71 (s, 1H) , 10.47 (s, 1H) , 9.60 (s, 2H) , 8.39 (d, J = 7.6 Hz, 1H) , 7.91 (s, 1H) , 7.71 –7.47 (m, 2H) , 7.31 –7.38 (m, 3H) , 7.06 (d, J = 12.4 Hz, 1H) , 6.64 (s, 1H) , 6.36 (s, 1H) , 5.70 –5.37 (m, 4H) , 4.69 (s, 1H) , 4.38 –4.29 (m, 2H) , 4.03 –3.92 (m, 2H) , 3.49 (s, 2H) , 3.30 –2.86 (m, 6H) , 2.69 (s, 1H) , 2.46 –1.96 (m, 17H) , 1.66 –1.73 (m, 4H) , 1.25 –1.39 (m, 4H) , 1.06 –0.66 (m, 16H) . LCMS (ESI) : RT = 0.909 min, m/z found 656.8 [M/2+H] ⁺.

Compound 30

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) -L-histidyl-L-valinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 14.25 (s, 1H) , 10.37 (s, 1H) , 9.78 –9.65 (m, 2H) , 8.93 (s, 2H) , 8.49 –8.29 (m, 4H) , 7.74 (d, J = 10.8 Hz, 1H) , 7.39 (d, J = 7.6 Hz, 4H) , 7.23 –7.10 (m, 2H) , 6.65 (s, 1H) , 6.35 (s, 1H) , 5.73 (d, 1H) , 5.55 (s, 2H) , 5.45 (s, 1H) , 5.14 (s, 1H) , 4.84 (s, 1H) , 4.41 (d, J = 6.4 Hz, 1H) , 4.03 –3.93 (m, 2H) , 3.62 (s, 3H) , 2.93-3.02 (m, 13H) , 2.69 (s, 1H) , 2.44 (s, 3H) , 2.15-2.30 (m, 6H) , 1.66-1.82 (m, 4H) , 1.25-1.36 (m, 3H) , 0.85-0.98 (m, 16H) . LCMS (ESI) : RT = 0.906 min, m/z found 656.8 [M/2+H] ⁺.

Compound 31

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (1, 1, 1-trifluoro-3-morpholinopropan-2-yl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate. ¹H NMR (400 MHz, DMSO) δ 10.29 (s, 1H) , 9.58 (d, J = 9.6 Hz, 1H) , 8.33 (s, 1H) , 7.70 (d, J = 10.8 Hz, 1H) , 7.36 (d, J = 8.4 Hz, 2H) , 7.29 (d, J = 8.4 Hz, 2H) , 7.12 (s, 1H) , 6.66 (s, 1H) , 6.35 (s, 1H) , 5.65 (d, J = 19.2 Hz, 1H) , 5.51 (s, 2H) , 5.39 (d, J = 19.2 Hz, 2H) , 4.72 (s, 2H) , 4.58 (s, 1H) , 4.40 (s, 2H) , 4.10 (d, J = 17.6 Hz, 2H) , 3.95 (d, J = 12.0 Hz, 2H) , 3.12 –2.96 (m, 3H) , 2.94 –2.86 (m, 3H) , 2.69 (s, 3H) , 2.61 (d, J = 8.8 Hz, 3H) , 2.37 (d, J = 14.4 Hz, 10H) , 2.22 –1.96 (m, 10H) , 1.93 (s, 1H) , 1.82 –1.70 (m, 4H) , 1.50 –1.34 (m, 4H) , 1.26 (s, 5H) , 0.95 –0.80 (m, 14H) . LCMS (ESI) : RT = 0.606 min, m/z found 616.7 [M/2+H] ⁺.

Compound 32

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5-hydroxy-4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-histidyl-D-valinate. LCMS (ESI) : RT = 2.071 min, m/z found 561.4 [M/2+H] ⁺.

Compound 33

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5-hydroxy-4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-histidyl-L-valinate. LCMS (ESI) : RT = 2.004 min, m/z found 561.4 [M/2+H] ⁺.

Compound 34

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl 1- ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carboxamido) cyclopropane-1-carboxylate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 9.78 –9.65 (m, 1H) , 8.65 (s, 1H) , 8.53 –8.47 (m, 1H) , 7.84 (d, J = 10.8 Hz, 1H) , 7.41 –7.37 (m, 4H) , 7.25 (s, 1H) , 6.66 (s, 1H) , 6.35 (s, 1H) , 5.75 –5.70 (m, 1H) , 5.52 (s, 2H) , 5.46 –5.41 (m, 1H) , 5.14 (s, 1H) , 3.99 –3.93 (m, 3H) , 3.63 –3.59 (m, 2H) , 3.09 –2.90 (m, 8H) , 2.25 –2.06 (m, 8H) , 1.93 –1.76 (m, 8H) , 1.54 –1.41 (m, 6H) , 1.26 –1.17 (m, 2H) , 1.08 –0.99 (m, 4H) , 0.92 (t, J = 7.2 Hz, 3H) , 0.85 (d, J = 6.4 Hz, 6H) . LCMS (ESI) : RT = 0.995 min, m/z found 580.2 [M/2+H] ⁺.

Compound 35

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (1- (2- (6- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5-hydroxy-4H-1, 2, 4-triazol-4-yl) -1H-indol-1-yl) ethyl) piperidine-4-carbonyl) -L-histidyl-L-valinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO-d₆) : δ 11.25 (s, 1H) , 9.65 (s, 1H) , 9.35 (s, 1H) , 8.83 (s, 1H) , 8.46 –8.40 (m, 2H) , 7.71 (d, J = 10.8 Hz, 1H) , 7.51 (d, J = 8.4 Hz, 1H) , 7.43 –7.38 (m, 1H) , 7.26 (t, J = 9.2 Hz, 2H) , 7.06 (s, 1H) , 6.83 (s, 1H) , 6.82 –6.77 (m, 1H) , 6.44 (d, J = 2.0 Hz, 1H) , 6.23 (s, 1H) , 5.77 –5.67 (m, 1H) , 5.46 –5.41 (m, 1H) , 5.12 (s, 1H) , 4.82 –4.87 (m, 1H) , 4.41 –4.37 (m, 1H) , 4.24 (s, 2H) , 3.71 (s, 2H) , 3.52 (s, 3H) , 3.22 –3.17 (m, 8H) , 2.68 –2.66 (m, 1H) , 2.52 (d, J = 1.6 Hz, 2H) , 2.41 (s, 3H) , 2.35 –2.28 (m, 2H) , 2.21 –2.12 (m, 3H) , 1.99 –1.64 (m, 5H) , 0.97 (d, J = 6.8 Hz, 3H) , 0.93 –0.86 (m, 12H) . LCMS (ESI) : RT = 0.852 min, m/z found 580.2 [M/2+H] ⁺.

Compound 36

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) benzoyl) -L-histidyl-L-valinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO-d₆) : δ 9.77 (s, 1H) , 9.65 (s, 1H) , 8.51-8.41 (m, 4H) , 7.89 (s, 2H) , 7.70 (d, J = 10.8 Hz, 1H) , 7.39 (d, J = 13.6 Hz, 6H) , 7.10 (d, J = 7.2 Hz, 1H) , 6.67 (s, 1H) , 6.32 (s, 1H) , 5.72 (d, J = 19.2 Hz, 1H) , 5.54 (s, 2H) , 5.44 (d, J = 20 Hz, 1H) , 5.12 (s, 1H) , 5.04 (d, J = 5.6 Hz, 1H) , 4.43-4.39 (m, 2H) , 4.00 –3.92 (m, 2H) , 3.45 –3.11 (m, 15H) , 2.73 –2.61 (m, 2H) , 2.52 (s, 2H) , 2.41 (s, 3H) , 2.36 –2.27 (m, 3H) , 2.16 (d, J = 7.4 Hz, 3H) , 0.97-0.85 (m, 15H) . LCMS (ESI) : RT = 0.870 min, m/z found 653.8 [M/2+H] ⁺.

Compound 37

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-histidyl-L-valinate. ¹H NMR (400 MHz, DMSO) δ 11.82 (s, 1H) , 10.52 –10.38 (m, 1H) , 9.58 (s, 2H) , 8.60 (s, 1H) , 7.94 (s, 1H) , 7.68 (d, J = 11.2 Hz, 2H) , 7.56 (s, 2H) , 7.30 (s, 12H) , 7.02 (s, 2H) , 6.84 (s, 2H) , 6.60 (s, 3H) , 6.34 (s, 3H) , 5.96 –5.82 (m, 2H) , 5.60 (s, 1H) , 5.50 (s, 4H) , 5.38 (s, 2H) , 4.75 (s, 2H) , 4.56 (s, 1H) , 4.34 (s, 3H) , 4.24 (s, 2H) , 3.96 (s, 7H) , 3.08 (s, 9H) , 3.02 –2.94 (m, 5H) , 2.86 (s, 8H) , 1.24 (s, 6H) , 1.15 (s, 5H) , 0.96 (s, 5H) , 0.94 (d, J = 4.4 Hz, 8H) , 0.92 (s, 8H) , 0.90 (s, 7H) , 0.88 (s, 3H) , 0.82 (s, 8H) , 0.80 (s, 10H) , 0.78 (s, 7H) . LCMS (ESI) : RT = 0.702 min, m/z found 615.8 [M/2+H] ⁺.

Compound 38

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) benzoyl) -L-histidyl-L-valinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 9.77 (s, 1H) , 9.65 (s, 1H) , 8.51-8.41 (m, 6H) , 7.89 (s, 2H) , 7.70-7.54 (m, 1H) , 7.39-7.20 (m, 7H) , 7.10-7.04 (m, 1H) , 6.67 (s, 1H) , 6.32 (s, 1H) , 5.72-5.64 (m, 1H) , 5.54 (s, 2H) , 5.44-5.32 (m, 1H) , 5.12 (s, 1H) , 5.04-4.90 (m, 1H) , 4.43-4.39 (m, 2H) , 4.00–3.92 (m, 2H) , 3.45–3.11 (m, 13H) , 2.52-2.40 (m, 5H) , 2.41- 2.31 (m, 3H) , 2.36–2.27 (m, 4H) , 1.16-1.04 (m, 3H) , 0.97-0.85 (m, 12H) . LCMS (ESI) : RT = 0.924 min, m/z found 653.8 [M/2+H] ⁺.

Compound 39

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (1- (2, 4-dihydroxy-5-isopropylphenyl) -5- (4- (morpholinomethyl) phenyl) -1H-1, 2, 3-triazole-4-carbonyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 9.73 –9.71 (m, 1H) , 9.02 (t, J = 6.0 Hz, 1H) , 8.52 –8.39 (m, 2H) , 7.90 –7.82 (m, 1H) , 7.42 –7.21 (m, 5H) , 7.03 –6.70 (m, 2H) , 6.38 –6.37 (m, 1H) , 5.92 –5.86 (m, 1H) , 5.76 –5.68 (m, 1H) , 5.52 (s, 2H) , 5.48 –5.40 (m, 1H) , 5.11 (s, 1H) , 4.31 –4.20 (m, 3H) , 4.00 –3.89 (m, 1H) , 3.56 –3.49 (m, 3H) , 3.14 –3.01 (m, 6H) , 2.26 –2.24 (m, 3H) , 2.18 –2.14 (m, 4H) , 1.24 (s, 1H) , 1.15 (s, 1H) , 1.00 (d, J = 6.4 Hz, 6H) , 0.89 (t, J = 7.2 Hz, 3H) . LCMS (ESI) : RT = 0.582 min, m/z found 913.0 [M+H] ⁺.

Compound 40

(1R, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H- 1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate. ¹H NMR (400 MHz, DMSO-d₆) : δ 10.46 (s, 1H) , 9.59 -9.56 (m, 1H) , 8.31 (s, 1H) , 7.66 (d, J = 10.8 Hz, 1H) , 7.36 -7.10 (m, 4H) , 7.10 (s, 1H) , 6.61 (s, 1H) , 6.34 (s, 1H) , 5.62 (d, J = 19.4 Hz, 1H) , 5.50 (s, 2H) , 5.32 (d, J = 19.2 Hz, 1H) , 4.36 (s, 1H) , 4.36 -4.08 (m, 1H) , 3.99 -3.91 (m, 3H) , 3.05 -3.02 (m, 2H) , 2.91 –2.86 (m, 1H) , 2.39 –2.02 (m, 18H) , 1.91 (s, 2H) , 1.78 -1.68 (m, 5H) , 1.46 –1.21 (m, 4H) , 0.93 -0.89 (m, 3H) , 0.81 -0.79 (m, 9H) . LCMS (ESI) : RT = 0.999 min, m/z found 567.4 [M/2+H] ⁺.

Compound 41

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-prolyl-L-valinate, 2, 2, 2-trifluoroacetate salt. ¹H NMR (400 MHz, DMSO) δ 9.76 (s, 1H) , 9.62 (t, J = 6.4 Hz, 1H) , 8.51 –8.44 (m, 4H) , 7.84 –7.71 (m, 1H) , 7.45 –7.36 (m, 4H) , 7.11 –7.06 (m, 1H) , 6.64 (s, 1H) , 6.33 –6.31 (m, 1H) , 5.75 –5.70 (m, 1H) , 5.57 –5.42 (m , 3H) , 5.10 (s, 1H) , 4.66 –4.60 (m, 1H) , 4.31 –4.26 (m, 2H) , 4.00 –3.91 (m, 3H) , 3.33 –3.22 (m, 6H) , 3.11 –2.87 (m, 8H) , 2.42 (s, 3H) , 2.27 –2.14 (m, 6H) , 2.05 –1.78 (m, 4H) , 1.00–1.09 (m, 9H) , 0.84 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 0.804 min, m/z found 595.8 [M/2+H] ⁺.

Compound 42

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-alanyl-L-prolyl-L-valinate. ¹H NMR (400 MHz, DMSO) δ 9.78 (s, 1H) , 9.65 –9.62 (m, 1H) , 8.79 –8.35 (m, 5H) , 7.77 –7.71 (m, 1H) , 7.49 –7.39 (m, 4H) , 7.13 –7.07 (m, 1H) , 6.68 (s, 1H) , 6.32 (s, 1H) , 5.74 –5.62 (m, 1H) , 5.58 –5.43 (m, 3H) , 5.11 (s, 1H) , 4.75 –4.47 (m, 2H) , 4.38 –4.16 (m, 2H) , 4.01 –3.92 (m, 4H) , 3.33 –3.23 (m, 4H) , 3.17 –3.09 (m, 4H) , 3.01 –2.90 (m, 6H) , 2.43 (s, 3H) , 2.21 –2.17 (m, 6H) , 2.03 –1.89 (m, 3H) , 1.29 –1.23 (m, 3H) , 1.01 –0.91 (m, 9H) , 0.87 (d, J = 6.4 Hz, 6H) . LCMS (ESI) : RT = 0.422 min, m/z found 1261.4 [M+H] ⁺.

Compound 43

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (1- ( (1- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperidin-4-yl) methyl) piperidine-4-carbonyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 10.10 (s, 1H) , 9.77 –9.75 (m, 1H) , 9.67 – 9.64 (m, 1H) , 9.25 –9.05 (m, 1H) , 8.52 –8.50 (m, 2H) , 7.84 (d, J = 10.8 Hz, 1H) , 7.56 –7.46 (m, 5H) , 7.17 –7.15 (m, 1H) , 6.69 (s, 1H) , 6.32 (s, 1H) , 5.74 –5.69 (m, 1H) , 5.58 –5.42 (m, 4H) , 5.11 (s, 1H) , 4.40 –3.93 (m, 8H) , 3.18 –3.05 (m, 4H) , 2.97 –2.90 (m, 8H) , 2.43 (s, 3H) , 2.17 –2.15 (m, 2H) , 1.95 –1.80 (m, 7H) , 1.43 –1.34 (m, 2H) , 1.03 –0.92 (m, 3H) , 0.87 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 1.196 min, m/z found 567.4 [M/2+H] ⁺.

Compound 44

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- ( (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindolin-5-yl) methyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 10.03 (s, 1H) , 9.64 (s, 1H) , 8.46 (s, 3H) , 8.29 (s, 1H) , 7.84 (d, J = 10.4 Hz, 1H) , 7.28 (s, 3H) , 7.16 (s, 1H) , 7.04 (s, 1H) , 6.40 (s, 1H) , 5.79 –5.64 (m, 1H) , 5.60 –5.34 (m, 3H) , 5.12 (s, 1H) , 4.78 (s, 4H) , 4.16 –3.97 (m, 2H) , 3.11 –2.94 (m, 13H) , 2.43 (s, 4H) , 2.20 –2.11 (m, 5H) , 1.75 (s, 4H) , 1.30 (m, 3H) , 1.14 (d, J = 6.8 Hz, 6H) , 0.91 (t, J = 7.2 Hz, 5H) . LCMS (ESI) : RT = 0.610 min, m/z found 1010.2 [M+H] ⁺.

Compound 45

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (S) -2- (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetamido) pentanoyl) -L-prolyl-L-valinate, 2, 2, 2-trifluoroacetate salt. ¹H NMR (400 MHz, DMSO-d₆) : δ 9.76 (s, 1H) , 9.64 (s, 1H) , 8.46 (s, 3H) , 8.38 -8.36 (m, 1H) , 7.72 (d, J = 10.4 Hz, 1H) , 7.40 (s, 4H) , 7.23 -7.07 (m, 1H) , 6.67 (s, 1H) , 6.32 -6.30 (m, 1H) , 5.72 -5.68 (m, 1H) , 5.59 –5.40 (m, 3H) , 5.11 (s, 1H) , 4.57 (s, 2H) , 4.28 –4.21 (m, 1H) , 4.06 –3.87 (m, 3H) , 3.71-3.63 (m, 3H) , 3.22 –2.84 (m, 11H) , 2.68 (s, 1H) , 2.43 (s, 3H) , 2.19 -2.17 (m , 5H) , 2.09 –1.83 (m, 4H) , 1.66 –1.25 (m, 5H) , 1.02 -0.94 (m, 10H) , 0.89 –0.79 (m, 9H) . LCMS (ESI) : RT = 0.814 min, m/z found 645.3 [M/2+H] ⁺.

Compound 46

(S) -4- ( (S) -2- ( ( (S) -1- ( ( (1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl) oxy) -3-methyl-1-oxobutan-2-yl) carbamoyl) pyrrolidin-1-yl) -3- (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetamido) -4-oxobutanoic acid--2, 2, 2-trifluoroacetic acid. ¹H NMR (400 MHz, DMSO-d₆) : δ 9.75 (s, 1H) , 9.64 (s, 1H) , 8.45 (s, 3H) , 8.23 -8.21 (m, 1H) , 7.73 -7.70 (m, 1H) , 7.41 (s, 4H) , 7.14 (s, 1H) , 6.68 (s, 1H) , 6.32 (s, 1H) , 5.72 -5.67 (m, 1H) , 5.52 -5.51 (m, 2H) , 5.47 -5.42 (m, 1H) , 5.10 (s, 1H) , 4.93 (s, 1H) , 4.21 -4.19 (m, 1H) , 4.13 -4.04 (m, 1H) , 3.98 -3.96 (m, 2H) , 3.69 (s, 2H) , 3.17 -3.07 (m, 8H) , 2.68 (s, 2H) , 2.43 (s, 4H) , 2.33 (s, 1H) , 2.20 -2.07 (m, 4H) , 1.97 -1.90 (m, 4H) , 1.63 -1.52 (m, 2H) , 1.24 -1.15 (m, 2H) , 1.03 (s, 1H) , 1.00 –0.74 (m, 18H) . LCMS (ESI) : RT = 0.767 min, m/z found 653.2 [M/2+H] ⁺.

Compound 47

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-histidyl-L-valinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 11.82 (s, 1H) , 10.52 –10.38 (m, 1H) , 9.58 (s, 2H) , 8.60 (s, 1H) , 7.94 (s, 1H) , 7.68 (d, J = 11.2 Hz, 2H) , 7.56 (s, 2H) , 7.30 (s, 12H) , 7.02 (s, 2H) , 6.84 (s, 2H) , 6.60 (s, 3H) , 6.34 (s, 3H) , 5.96 –5.82 (m, 2H) , 5.60 (s, 1H) , 5.50 (s, 4H) , 5.38 (s, 2H) , 4.75 (s, 2H) , 4.56 (s, 1H) , 4.34 (s, 3H) , 4.24 (s, 2H) , 3.96 (s, 7H) , 3.08 (s, 9H) , 3.02 –2.94 (m, 5H) , 2.86 (s, 8H) , 1.24 (s, 6H) , 1.15 (s, 5H) , 0.96 (s, 5H) , 0.94 (d, J = 4.4 Hz, 8H) , 0.92 (s, 8H) , 0.90 (s, 7H) , 0.88 (s, 3H) , 0.82 (s, 8H) , 0.80 (s, 10H) , 0.78 (s, 7H) . LCMS (ESI) : RT = 1.265 min, m/z found 1230.8 [M+H] ⁺.

Compound 48

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) acetyl) -L-histidyl-L-prolyl-L-valinate, 2, 2, 2-trifluoroacetate salt. ¹H NMR (400 MHz, DMSO) δ 14.12 (s, 1H) , 10.20 (s, 1H) , 9.78 (s, 1H) , 9.65 (t, J = 6.0 Hz, 1H) , 8.90 (s, 1H) , 8.65 (d, J = 8.8 Hz, 1H) , 8.47 (s, 3H) , 7.67 (d, J = 10.8 Hz, 1H) , 7.47 –7.39 (m, 5H) , 7.09 (s, 1H) , 6.67 (s, 1H) , 6.32 (s, 1H) , 5.76 –5.69 (m, 1H) , 5.59 –5.42 (m, 3H) , 5.10 (s, 1H) , 4.94 –4.89 (m, 1H) , 4.60 –4.58 (m, 1H) , 4.39 –4.35 (m, 1H) , 4.01 –3.92 (m, 2H) , 3.78 –3.73 (m, 1H) , 3.64 –3.60 (m, 1H) , 3.15 –3.06 (m, 7H) , 3.01 –2.90 (m, 6H) , 2.42 (s, 3H) , 2.34 –2.28 (m, 2H) , 2.26 –2.15 (m, 5H) , 2.01 –1.90 (m, 4H) , 1.00 –0.92 (m, 12H) , 0.87 (d, J = 6.4 Hz, 6H) . LCMS (ESI) : RT = 0.602 min, m/z found 664.5 [M/2+H] ⁺.

Compound 49

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3-carbamoyl-5- (2, 4-dihydroxy-5-isopropylphenyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 9.76 (s, 1H) , 8.47 (s, 2H) , 8.33 –8.25 (m, 2H) , 7.84 (d, J = 10.8 Hz, 1H) , 7.71 (s, 1H) , 7.42 –7.33 (m, 4H) , 7.16 (s, 1H) , 6.59 (s, 1H) , 6.33 (s, 1H) , 5.75–5.68 (m, 1H) , 5.52 (d, J = 3.6 Hz, 2H) , 5.68–5.28 (m, 1H) , 5.11 (s, 1H) , 4.17 –4.12 (m, 1H) , 4.02 –3.97 (m, 1H) , 3.31 –3.23 (m, 5H) , 2.95 –2.87 (m, 5H) , 2.73 (s, 1H) , 2.69 –2.65 (m, 1H) , 2.53 (s, 3H) , 2.42 (s, 4H) , 2.35 –2.31 (m, 1H) , 2.20 –2.06 (m, 5H) , 1.76 (s, 6H) , 1.40 –1.34 (m, 3H) , 0.91 (t, J = 7.2 Hz, 3H) , 0.82 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 1.236 min, m/z found 526.4 [M/2+H] ⁺.

Compound 50

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl (2- (4- ( (2- (2, 4-dihydroxy-5-isopropylbenzoyl) isoindolin-5-yl) methyl) piperazin-1-yl) acetyl) -L-histidyl-L-valinate 2, 2, 2-trifluoroacetate. ¹H NMR (400 MHz, DMSO) δ 14.19 (s, 1H) , 10.04 (s, 1H) , 9.68 (s, 1H) , 8.83 (s, 1H) , 8.65 –8.51 (m, 4H) , 7.65 (d, J = 10.4 Hz, 1H) , 7.44 –7.29 (m, 3H) , 7.17 –7.03 (m, 3H) , 6.41 (s, 1H) , 5.73 –5.65 (m, 1H) , 5.59 –5.41 (m, 3H) , 5.09 (s, 1H) , 4.86 –4.80 (m, 5H) , 4.39 (t, J = 7.2 Hz, 1H) , 4.27 –4.00 (m, 2H) , 3.32 –3.24 (m, 5H) , 3.17 –3.06 (m, 6H) , 2.99 –2.93 (m, 4H) , 2.41 (s, 3H) , 2.33 –2.29 (m, 2H) , 2.23 –2.19 (m, 4H) , 1.13 (d, J = 6.4 Hz, 6H) , 0.98 –0.92 (m, 3H) , 0.89 (d, J = 6.8 Hz, 6H) . LCMS (ESI) : RT = 0.563 min, m/z found 554.3 [M/2+H] ⁺.

Compound 51

(1S, 9S) -1-amino-9-ethyl-5-fluoro-4-methyl-10, 13-dioxo-2, 3, 9, 10, 13, 15-hexahydro-1H, 12H-benzo [de] pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl ( (1r, 4S) -4- ( (4- (4- (3- (2, 4-dihydroxy-5-isopropylphenyl) -5- ( (2, 2, 2-trifluoroethyl) carbamoyl) -4H-1, 2, 4-triazol-4-yl) benzyl) piperazin-1-yl) methyl) cyclohexane-1-carbonyl) glycinate formate. ¹H NMR (400 MHz, DMSO-d₆) δ 10.45 (s, 1H) , 9.60 -9.56 (m, 1H) , 8.31 -8.30 (m, 1H) , 7.70 -7.68 (m, 1H) , 7.36 -7.29 (m, 4H) , 7.11 (s, 1H) , 6.61 (s, 1H) , 6.34 (s, 1H) , 5.65 –5.37 (m, 4H) , 4.39 (s, 1H) , 4.11 -4.10 (m, 1H) , 3.99 –3.91 (m, 3H) , 3.07 (s, 2H) , 2.92 –2.86 (m, 1H) , 2.75 –2.58 (m, 1H) , 2.41 –2.24 (m, 12H) , 2.17 –2.01 (m, 8H) , 1.86 –1.56 (m, 5H) , 1.40 -1.31 (m, 4H) , 0.92 -0.88 (m, 3H) , 0.81 -0.80 (m, 8H) . LCMS (ESI) : RT = 1.188 min, m/z found 1134.6 [M+H] ⁺.

Biological Assays

Cell lines

The following cancer cell lines were employed: ; MV4-11 human biphenotypic B myelomonocytic leukemia (ATCC, #CRL-9591) ; HCT-116 human colon carcinoma (ATCC, #CRL-247) . Cell lines were cultured essentially according to ATCC recommendations.

HSP90α-binding fluorescent polarization (FP) assay

Binding of test compounds to HSP90α (HSP90AA1) protein was measured by fluorescent polarization (FP) using the HSP90α (N-terminal) Assay Kit (BPS Bioscience, #50298) , following the manufacturer’s instructions, except as noted. Fluorescently labeled HSP90-binding compound FITC-geldanamycin (5 nM final concentration) was provided and employed. A 2.5-fold serial dilution of each test compound ranging from 20 μM to 5.2 nM was assayed for binding to HSP90α. After the final step of adding HSP90a protein to each assay well, plates were mixed by brief shaking, incubated at 25℃ for 120 min, and fluorescence was measured using a PerkinElmer EnVision Plate Reader. Background-subtracted mP values were calculated from raw data and a four-parameter “log [inhibitor] vs. response” curve was fitted and IC50 values (the concentration at which 50%of the maximal inhibition occurs) calculated using GraphPad Prism 7 software. Results are shown in Table 1.

Table 1

Reagent cancer cell line proliferation assays

Cells were plated in 96-well tissue culture plates at 4,000 cells/well and incubated at 37℃/5%CO₂ for 24-48 hr in 100 μl of media. 3-fold serial dilutions of each test compound were prepared ranging from 20 μM to 1.02 nM. MV4-11 cells (biphenotypic B-myelomonocytic leukemia) , and HCT116 (human colorectal carcinoma cell line) were then treated with test compounds at various concentrations with a final concentration of 0.5%DMSO/well, and then incubated at 37℃/5%CO₂ for 24 hr. 100 μl ofReagent (Promega Corporation, Madison, WI) was added to each well and processed according manufacturer’s protocol. Results were analyzed and IC50-values were calculated in GraphPad 7 software. Results are shown in Table 2. IC50<100 nM; B. IC50=100-1000 nM; C. IC50>1000 nM.

Table 2

Plasma Stability

Test compounds were prepared in DMSO at 1 mM and 1 mM propantheline or mevinolin working solutions were prepared in acetonitrile. Propantheline served as a positive control for human, dog and mouse plasma, while mevinolin was used a positive control for rat plasma. Assay plates were prewarmed at 37℃ for 15 minutes before adding 398 μL of plasma. Then 2 μL of 1mM test compound and 2 μL of 1mM control compound were added to 398 μL of plasma to reach a final concentration of 5 μM for the test compound and 5 μM for the control compound. The final concentration of organic solvents was 0.5 %. Each assay was performed in duplicate.

Plasma samples were incubated at 37℃. Aliquots of 50 μL of each plasma sample were then taken at 0, 1, 2, 4, 8 and 24 hours (Cpd 1-Cpd 10) or at 0, 15, 30, 60 and 120 min (Cpd 11-Cpd 26) . The reaction was halted by the addition of 450 μL cold methanol with 5%formic acid containing internal standards. All samples were vortexed for 10 minutes, followed by centrifugation at 3, 220 g for 30 minutes to precipitate proteins. From each sample 100 μL of supernatant was transferred to a new plate. Sample supernatants were diluted with ultrapure water and loaded on to a liquid chromatography mass spectrometer. Samples were analyzed by LC-MS/MS. All calculations were carried out using Microsoft Excel. Peak area ratios were determined from extracted ion chromatograms.

Percent compounds remaining at each time point were calculated by the following equation: Remaining Percentage t min (%) = Peak Area Ratio t min/Peak Area Ratio 0 min ′100. Where Peak Area Ratio t min is peak area ratio of control and test compounds at t min; Peak Area Ratio 0 min is peak area ratio of control and test compounds at zero time point. The slope value, k, was determined by linear regression of the natural logarithm of the remaining percentage of the parent drug vs. incubation time curve. The in vitro half-life (in vitro t1/2) was determined from the slope value: In vitro t_1/2 = - (0.693 /k) . Results are shown in Table 3.

Table 3

Note: Cpd 1-Cpd 10 were incubated at 0, 1, 2, 4, 8 and 24 hours Cpd 11-Cpd 26 at 0, 15, 30, 60 and 120 min

Modifications and variations of the described methods and compositions of the present disclosure will be apparent to those skilled in the art without departing from the scope and spirit of the disclosure. Although the disclosure has been described in connection with specific embodiments, it should be understood that the disclosure as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the disclosure are intended and understood by those skilled in the relevant field in which this disclosure resides to be within the scope of the disclosure as represented by the following claims.

INCORPORATION BY REFERENCE

All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication was specifically and individually indicated to be incorporated by reference.

Claims

A compound of the Formula I:

or a pharmaceutically acceptable salt thereof, wherein

R^Q and R^T are each independently selected from H, optionally substituted (C₁-C₄) alkyl, optionally substituted aryl, and optionally substituted heteroaryl, or R^Q and R^T are taken together with the carbon they are attached to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl;

X is absent or -NR^W, wherein R^W is H or (C₁-C₄) alkyl;

L is a linker; and

A is a chemical moiety that binds HSP90 protein.
The compound of Claim 1, wherein the compound is of the Formula II or III:

or a pharmaceutically acceptable salt thereof.
The compound of Claim 1 or 2, wherein the compound is of the Formula III:

or a pharmaceutically acceptable salt thereof.
The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein X is absent or -NR^W, wherein R^W is H or CH₃.
The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein X is absent or NH.
The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein R^Q and R^T are each independently selected from H and (C₁-C₄) alkyl.
The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein R^Q and R^T are each independently selected from H, CH (CH₃) ₂, and CH₃.
The compound of any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein R^Q is H and R^T is selected from H, CH (CH₃) ₂, and CH₃.
The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein R^Q and R^T are each H.
The compound of any one of Claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein A is selected from

Z and Z’ are each independently selected from N or CH;

R¹, R^1A, and R^1AA are each independently selected from (C₁-C₄) alkyl, halo (C₁-C₄) alkyl, halo, (C₁-C₄) alkoxy, and halo (C₁-C₄) alkoxy;

R³, R^3A, and R^3AA are each independently selected from hydrogen, halo, (C₁-C₄) alkyl, halo (C₁-C₄) alkyl, (C₁-C₄) alkoxy, halo (C₁-C₄) alkoxy, - (C₀-C₄) alkylheterocyclyl, and - (C₀-C₄) heteroaryl, wherein said heterocyclyl and heteroaryl are each optionally substituted with 1 to 3 groups selected from R^Z;

R^Z is is selected from (C₁-C₄) alkyl, halo (C₁-C₄) alkyl, halo, (C₁-C₄) alkoxy, and halo (C₁-C₄) alkoxy;

R^a and R^b are each independently selected from hydrogen, (C₁-C₄) alkyl, and halo (C₁-C₄) alkyl;

R⁴ is (C₁-C₄) alkyl;

R⁵ and R^5A are each independently (C₁-C₄) alkyl;

R⁶ and R^6A are each independently hydrogen or (C₁-C₄) alkyl;

R⁷ is halo; and

the wavy line indicates the point of attachment to linker L.
The compound of any one of Claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein A is selected from
The compound of Claim 10 or 11, or a pharmaceutically acceptable salt thereof, wherein Z and Z’ are each CH.
The compound of any one of Claims 10 to 12, or a pharmaceutically acceptable salt thereof, wherein R¹, R^1A, and R^1AA are each (C₁-C₄) alkyl.
The compound of any one of Claims 10 to 13, or a pharmaceutically acceptable salt thereof, wherein R¹, R^1A, and R^1AA are each isopropyl.
The compound of any one of Claims 10 to 14, or a pharmaceutically acceptable salt thereof, wherein R³, R^3A, and R^3AA are each independently selected from hydrogen, - (C₁-C₄) alkylheterocyclyl, and heteroaryl, wherein said heterocyclyl and heteroaryl are each optionally substituted with 1 to 3 groups selected from R^Z.
The compound of any one of Claims 10 to 15, or a pharmaceutically acceptable salt thereof, wherein R^3A is hydrogen.
The compound of any one of Claims 10 to 16, or a pharmaceutically acceptable salt thereof, wherein R^3AA is hydrogen.
The compound of any one of Claims 10 to 17, or a pharmaceutically acceptable salt thereof, wherein R³ is selected from - (C₁-C₄) alkyl (5-to 6-membered) heterocyclyl and fused 8-10 membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R^Z.
The compound of any one of Claims 10 to 18, or a pharmaceutically acceptable salt thereof, wherein R³ is selected from - (C₁-C₄) morpholinyl, indolyl, piperazinyl, wherein said morpholinyl, indolyl, and piperazinyl are each optionally substituted with 1 to 3 groups selected from R^Z.
The compound of any one of Claims 10 to 19, or a pharmaceutically acceptable salt thereof, wherein R^Z is (C₁-C₄) alkyl.
The compound of any one of Claims 10 to 20, or a pharmaceutically acceptable salt thereof, wherein R^a is hydrogen; and R^b is halo (C₁-C₄) alkyl.
The compound of any one of Claims 10 to 21, or a pharmaceutically acceptable salt thereof, wherein R^a is hydrogen; and R^b is CH₂CF₃.
The compound of any one of Claims 10 to 22, or a pharmaceutically acceptable salt thereof, wherein R⁴ is CH₃.
The compound of any one of Claims 10 to 23, or a pharmaceutically acceptable salt thereof, wherein R⁵ and R^5A are each CH₃.
The compound of any one of Claims 10 to 24, or a pharmaceutically acceptable salt thereof, wherein R⁶ and R^6A are each hydrogen.
The compound of any one of Claims 10 to 25, or a pharmaceutically acceptable salt thereof, wherein R⁷ is chloro.
The compound of any one of Claims 1 to 26, or a pharmaceutically acceptable salt thereof, wherein

L is selected from - (CH₂) _r-HET¹-X¹-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹- (CH₂) _q-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET²- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-HET²- (CH₂) _s-C (O) - (CH₂) _m-X²-, - (CH₂) _yOC (O) -, - (CH₂) _r-HET¹- (CH₂) _w-S-S-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET²- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET³-OC (O) -, - (CH₂) _r-HET²-X¹-HET³-OC (O) -, - (CH₂) _r-HET³-X¹- (CH₂) _s-OC (O) -, - (CH₂) _r-HET³-X¹-HET²-OC (O) -, -X¹-HET³-OC (O) -, -X¹-HET³-X²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹- (CH₂) _q-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹- (CH₂) _w-S-S-HET²- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-HET²- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _q-HET¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-HET¹- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²- (CH₂) _w-S-S-HET¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-C (O) - (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _q-HET¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-X¹- (CH₂) _q-HET³- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET³- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET¹-HET³- (CH₂) _s-C (O) - (CH₂) _m-X²-, - (CH₂) _r-HET¹- (CH₂) _w-S-S-HET³- (CH₂) _s-OC (O) -, - (CH₂) _r-HET¹-X¹-HET³- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, - (CH₂) _r-HET³-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET²-X¹-HET³- (CH₂) _s-OC (O) -, - (CH₂) _rNR^f (CH₂) _v-HET³-X¹-HET¹- (CH₂) _s-C (O) O- (CH₂) _m-X²-, - (CH₂) _rNR^f (CH₂) _v-HET³- (CH₂) _s-NHC (O) O- (CH₂) _m-X²-, -C (O) NH-CRR’-C (O) NH-CRR’-C (O) NHCRR’C (O) NH (CH₂) _m-X²-, -C (O) NH-CRR’-C (O) NH-CRR’-C (O) NH-HET²- (CH₂) _s-OC (O) -, C (O) , -C (O) -X¹-HET¹-X²-, -X¹-HET¹-X², -C (O) -HET¹-X¹-HET²-X², and C (O) -X¹-O-;

m, q, r, s, v, w, and y are each independently 0 to 4;

R^f is selected from hydrogen and (C₁-C₄) alkyl;

X¹ and X² are each independently selected from CO and –CH₂-;

HET¹, HET², and HET³ are each independently selected from monocyclic heterocyclyl, bicyclic heterocyclyl, monocyclic cycloalkyl, and phenyl; and

R’ and R” are each independently selected from hydrogen and (C₁-C₄) alkyl, wherein said (C₁-C₄) alkyl is optionally substituted with phenyl or NHC (O) NH₂.
The compound of any one of Claims 1 to 27, or a pharmaceutically acceptable salt thereof, wherein L is selected from C (O) , -C (O) -X¹-HET¹-X²-, -X¹-HET¹-X², -C (O) -HET¹-X¹-HET²-X², and C (O) -X¹-O-.
The compound of Claim 27 or 28, or a pharmaceutically acceptable salt thereof, wherein X¹ and X² are each –CH₂-.
The compound of any one of Claims 27 to 29, or a pharmaceutically acceptable salt thereof, wherein HET¹, HET², and HET³ are each independently selected from monocyclic heterocyclyl and monocyclic cycloalkyl.
The compound of any one of Claims 27 to 30, or a pharmaceutically acceptable salt thereof, wherein HET¹ and HET², and HET³ are each independently selected from piperizinyl and cyclohexyl.
The compound of any one of Claims 27 to 30, or a pharmaceutically acceptable salt thereof, wherein L is selected from C (O) , and C (O) CH₂O*, wherein *indicates the point of attachment to A.
The compound of Claim 1, wherein the compound is selected from

or pharmaceutically acceptable salt of any of the foregoing.
A pharmaceutical composition comprising a compound of any one of Claims 1 to 33, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.
A method of treating cancer comprising administering to a subject a therapeutically effective amount of a compound of any one of Claims 1 to 33, or a pharmaceutically acceptable salt thereof, or the composition of Claim 34.