WO2022032284A1 - Macrocyclic compounds - Google Patents

Macrocyclic compounds Download PDF

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Publication number
WO2022032284A1
WO2022032284A1 PCT/US2021/071102 US2021071102W WO2022032284A1 WO 2022032284 A1 WO2022032284 A1 WO 2022032284A1 US 2021071102 W US2021071102 W US 2021071102W WO 2022032284 A1 WO2022032284 A1 WO 2022032284A1
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WO
WIPO (PCT)
Prior art keywords
cancer
compound
methyl
mmol
lcms
Prior art date
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PCT/US2021/071102
Other languages
French (fr)
Inventor
Junhu Zhang
Peter Qinhua HUANG
Kevin Duane BUNKER
Sobhana Babu Boga
Sunny Abraham
Brant Clayton Boren
Wanlong Jiang
Sunil Paliwal
Original Assignee
Zeno Management, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication of WO2022032284A1 publication Critical patent/WO2022032284A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D515/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D515/22—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains four or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the present application relates to compounds that are Mcl-1 inhibitors and methods of using them to treat conditions characterized by excessive cellular proliferation, such as cancer.
  • Mcl-1 myeloid cell leukemia-1
  • Bcl-2 family of proteins. MCL-1 is widely expressed in human tissues and is primarily located in the mitochondria in cells. Upregulation of Mcl-1 occurs in different cancer types. Additionally, overexpression of Mcl-1 has been linked to drug resistance to several cancer therapies.
  • Some embodiments provide a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • compositions that can include an effective amount of one or more of compounds of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.
  • Some embodiments described herein relate to a method for ameliorating and/or treating a cancer described herein that can include administering an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof)
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein.
  • Some embodiments described herein relate to a method for inhibiting replication of a malignant growth or a tumor that can include contacting the growth or the tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the malignant growth or tumor is due to a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • Some embodiments described herein relate to a method for inhibiting the activity of Mcl-1 in a cell that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof)
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of Mcl-1.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of Mcl- 1.
  • Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include inhibiting the activity of Mcl-1 using an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof).
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof).
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
  • Mcl-1 Myeloid Cell Leukemia 1
  • Mcl-1 Myeloid Cell Leukemia 1
  • Amplification of the MCL1 gene and/or overexpression of the Mcl-1 protein has been observed in multiple cancer types and is commonly implicated in tumor development.
  • MCL1 is one of the most frequently amplified genes in human cancers.
  • Mcl-1 is a critical survival factor and it has been shown to mediate drug resistance to a variety of anti-cancer agents. Mcl-1 promotes cell survival by binding to pro-apoptotic proteins like Bim, Noxa, Bak, and Bax and neutralizing their deathinducing activities.
  • Ca to Cb in which “a” and “b” are integers refer to the number of carbon atoms in a group.
  • the indicated group can contain from “a” to “b”, inclusive, carbon atoms.
  • a “C 1 to C 4 alkyl” group refers to all alkyl groups having from 1 to 4 carbons, that is, CH3-, CH3CH2-, CH3CH2CH2-, (CH3)2CH-, CH3CH2CH2CH2-, CH3CH2CH(CH3)- and (CH3)3C-. If no “a” and “b” are designated, the broadest range described in these definitions is to be assumed.
  • alkyl refers to a fully saturated aliphatic hydrocarbon group.
  • the alkyl moiety may be branched or straight chain.
  • branched alkyl groups include, but are not limited to, iso-propyl, sec-butyl, iso-butyl, t-butyl and the like.
  • straight chain alkyl groups include, but are not limited to, methyl, ethyl, n-propyl and n-butyl.
  • An alkyl group may be substituted or unsubstituted.
  • halogen atom or “halogen” as used herein, means any one of the radio-stable atoms of column 7 of the Periodic Table of the Elements, such as, fluorine, chlorine, bromine and iodine.
  • substituents e.g. haloalkyl
  • substituents there may be one or more substituents present.
  • haloalkyl may include one or more of the same or different halogens.
  • C 1 -C 3 alkoxyphenyl may include one or more of the same or different alkoxy groups containing one, two or three atoms.
  • a radical indicates species with a single, unpaired electron such that the species containing the radical can be covalently bonded to another species.
  • a radical is not necessarily a free radical. Rather, a radical indicates a specific portion of a larger molecule.
  • the term “radical” can be used interchangeably with the term “group.”
  • pharmaceutically acceptable salt refers to a salt of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. In some embodiments, the salt is an acid addition salt of the compound.
  • compositions can be obtained by reacting a compound with inorganic acids such as hydrohalic acid (e.g., hydrochloric acid or hydrobromic acid), a sulfuric acid, a nitric acid and a phosphoric acid (such as 2,3-dihydroxypropyl dihydrogen phosphate).
  • hydrohalic acid e.g., hydrochloric acid or hydrobromic acid
  • sulfuric acid e.g., sulfuric acid
  • nitric acid e.g., 1,3-dihydroxypropyl dihydrogen phosphate
  • Pharmaceutical salts can also be obtained by reacting a compound with an organic acid such as aliphatic or aromatic carboxylic or sulfonic acids, for example formic, acetic, succinic, lactic, malic, tartaric, citric, ascorbic, nicotinic, methanesulfonic, ethanesulfonic, p- toluenesulfonic, trifluoro acetic, benzoic, salicylic, 2-oxopentanedioic or naphthalenesulfonic acid.
  • an organic acid such as aliphatic or aromatic carboxylic or sulfonic acids
  • Pharmaceutical salts can also be obtained by reacting a compound with a base to form a salt such as an ammonium salt, an alkali metal salt, such as a sodium, a potassium or a lithium salt, an alkaline earth metal salt, such as a calcium or a magnesium salt, a salt of a carbonate, a salt of a bicarbonate, a salt of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C1-C7 alkylamine, cyclohexylamine, triethanolamine, ethylenediamine and salts with amino acids such as arginine and lysine.
  • a salt such as an ammonium salt, an alkali metal salt, such as a sodium, a potassium or a lithium salt, an alkaline earth metal salt, such as a calcium or a magnesium salt, a salt of a carbonate, a salt of a bicarbonate, a salt of organic bases such as di
  • a salt is formed by protonation of a nitrogen-based group (for example, NH2)
  • the nitrogen-based group can be associated with a positive charge (for example, NH2 can become NH3 + ) and the positive charge can be balanced by a negatively charged counterion (such as Cl ).
  • each center may independently be of R-configuration or S -configuration or a mixture thereof.
  • the compounds provided herein may be enantiomerically pure, enantiomerically enriched, racemic mixture, diastereomerically pure, diastereomerically enriched or a stereoisomeric mixture.
  • each double bond may independently be E or Z a mixture thereof.
  • all tautomeric forms are also intended to be included.
  • valencies are to be filled with hydrogens or isotopes thereof, e.g., hydrogen- 1 (protium) and hydrogen-2 (deuterium).
  • each chemical element as represented in a compound structure may include any isotope of said element.
  • a hydrogen atom may be explicitly disclosed or understood to be present in the compound.
  • the hydrogen atom can be any isotope of hydrogen, including but not limited to hydrogen- 1 (protium) and hydrogen-2 (deuterium).
  • reference herein to a compound encompasses all potential isotopic forms unless the context clearly dictates otherwise.
  • the methods and combinations described herein include crystalline forms (also known as polymorphs, which include the different crystal packing arrangements of the same elemental composition of a compound), amorphous phases, salts, solvates and hydrates.
  • the compounds described herein exist in solvated forms with pharmaceutically acceptable solvents such as water, ethanol or the like.
  • the compounds described herein exist in unsolvated form.
  • Solvates contain either stoichiometric or non- stoichiometric amounts of a solvent, and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol or the like. Hydrates are formed when the solvent is water or alcoholates are formed when the solvent is alcohol.
  • the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein.
  • the term “comprising” is to be interpreted synonymously with the phrases “having at least” or “including at least”.
  • the term “comprising” means that the compound, composition or device includes at least the recited features or components, but may also include additional features or components.
  • R 1 can be fluoro, chloro or methyl
  • R 2 can be hydrogen or fluoro
  • Ring A can be n alkyl
  • Ring B can , wherein R 6 and R 7 can be independently hydrogen or an unsubs
  • the ormula (I) can be substituted with a halogen or an unsubstituted alkyl.
  • R 1 can be fluoro, such that the indole can have er re o- substituted.
  • R 2 can be hydrogen.
  • R 2 When the naphthyl ring is fluoro-substituted R 2 can be fluoro.
  • R 2 can be fluoro.
  • Having a fluoro on the naphthyl ring can be advantageous. Examples of advantages include, but are not limited to, the following: increased potency, greater stability and/or more efficient interaction with the Mcl-1 protein.
  • By having a fluoro-substituted naphthyl there may be increased hydrophobic interaction between the compound and Mcl-1 protein (for example, Valine 274, Leucine 290 and/or isoleucine 294), and thus, the potency of a compound described herein (including pharmaceutically acceptable salts thereof) can be increased.
  • the fluoro on the naphthyl can act as an electron-withdrawing group and mitigate the electronic density of the sulfur and/or oxygen attached to the naphthyl which may lead to more efficient compound/Mcl- 1 protein hydrophobic interaction and/or more metabolic stability.
  • the fluoro on the naphthyl may block potential metabolic soft spots of the compound, and result in more stable compounds.
  • Ring B can be a pyrazole ring.
  • the pyrazole ring can be unsubstituted or alkyl-substituted.
  • Ring B can be .
  • Ring B can be .
  • the pyrazole ring can be unsubstituted, for example, then R 6 is hydrogen and/or R 7 is hydrogen. Also as described herein, the pyrazole ring can be alkylsubstituted. The pyrazole ring can be alkyl-substituted when R 6 and/or R 7 is an unsubstituted Ci- 4 alkyl. Examples of an unsubstituted Ci-4 alkyls include methyl, ethyl, n-propyl, iso-propyl, n- butyl, sec-butyl, iso-butyl and tert-butyl. Exemplary Ring B moieties include the following:
  • Ring A can be a monocyclic nitrogen-containing ring or a bicyclic nitrogencontaining ring.
  • Ring A can be a pyrazole ring, such The substitution on Ring A can vary.
  • R 3 and R 4 combinations for Ring A include the following: R 3 /R 4 R 3 /R 4 R 3 /R 4 R 3 /R 4 meth l/meth l meth l/eth l meth l/n-ro l meth l/iso- ro l
  • Ring A can also be a bicyclic nitrogen-containing ring.
  • the ring fused to the pyrazole ring of Ring A can be a five-membered carbocyclyl or a six-membered carbocyclyl. In some embodiments, Ring A ca . In some embodiments, Ring A can be s, e embodiments, Ring A can be . Ring A can be fused to a six-membered In some embodiments, Ring A can be.
  • R 5 can be an unsubstituted C1-4 alkyl. Example of suitable are described herein.
  • R can be methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl and tert-butyl.
  • Examples of compounds that can be used in a method and/or use described herein, along with pharmaceutically acceptable salts thereof, include the following:
  • R 6 cannot be methyl.
  • R 7 cannot be methyl.
  • R 1 cannot be chloro.
  • R 2 cannot be hydrogen.
  • R 2 cannot be fluoro.
  • R 1 cannot be chloro when R 6 is methyl.
  • R 2 when R 2 is fluoro; then R 1 cannot be chloro. In some embodiments, when R 2 is hydrogen; and R 6 is methyl; then R 1 cannot be chloro. In other embodiments, when R 2 is fluoro; and R 6 is methyl; then R 1 cannot be chloro.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof cannot a compound disclosed in WO 2018/178226 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof cannot a compound disclosed in WO 2017/181625 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof cannot a compound disclosed in WO 2020/103864 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof cannot a compound disclosed in WO 2020/151738 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • Compounds of the Formula (I), or pharmaceutically acceptable salts thereof can be made in various ways by those skilled using known techniques as guided by the detailed teachings provided herein.
  • compounds of the Formula (I) are prepared in accordance with General Scheme 1 as shown herein.
  • Compounds of Formula (I), and pharmaceutically acceptable salts thereof can be prepared according to the preparation shown in Scheme 1.
  • Compound A can undergo a Mitsunobu reaction and close the ring to form the macrocyclic Compound B.
  • P represents a suitable protecting group. Removal of the protecting group via a hydrolysis reaction provides a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • compositions that can include an effective amount of one or more compounds described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.
  • composition refers to a mixture of one or more compounds and/or salts disclosed herein with other chemical components, such as diluents or carriers.
  • the pharmaceutical composition facilitates administration of the compound to an organism.
  • Pharmaceutical compositions can also be obtained by reacting compounds with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid.
  • Pharmaceutical compositions will generally be tailored to the specific intended route of administration.
  • physiologically acceptable defines a carrier, diluent or excipient that does not abrogate the biological activity and properties of the compound nor cause appreciable damage or injury to an animal to which delivery of the composition is intended.
  • a “carrier” refers to a compound that facilitates the incorporation of a compound into cells or tissues.
  • DMSO dimethyl sulfoxide
  • DMSO dimethyl sulfoxide
  • a “diluent” refers to an ingredient in a pharmaceutical composition that lacks appreciable pharmacological activity but may be pharmaceutically necessary or desirable.
  • a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and/or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation.
  • a common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the pH and isotonicity of human blood.
  • an “excipient” refers to an essentially inert substance that is added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the composition.
  • stabilizers such as anti-oxidants and metal-chelating agents are excipients.
  • the pharmaceutical composition comprises an anti-oxidant and/or a metal-chelating agent.
  • a “diluent” is a type of excipient.
  • compositions disclosed herein may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes. Additionally, the active ingredients are contained in an amount effective to achieve its intended purpose. Many of the compounds used in the pharmaceutical combinations disclosed herein may be provided as salts with pharmaceutically compatible counterions.
  • a compound, salt and/or composition can be administered orally.
  • the liposomes will be targeted to and taken up selectively by the organ. For example, intranasal or pulmonary delivery to target a respiratory disease or condition may be desirable.
  • compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient.
  • the pack may for example comprise metal or plastic foil, such as a blister pack.
  • the pack or dispenser device may be accompanied by instructions for administration.
  • the pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert.
  • Compositions that can include a compound and/or salt described herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
  • Some embodiments described herein relate to a method for ameliorating and/or treating a cancer described herein that can include administering an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof)
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein.
  • Some embodiments described herein relate to a method for inhibiting replication of a malignant growth or a tumor that can include contacting the growth or the tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the malignant growth or tumor is due to a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
  • Some embodiments described herein relate to a method for inhibiting the activity of Mcl-1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof)
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of Mcl-1.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of Mcl-1.
  • Some embodiments described herein relate to a method for inhibiting the activity of Mcl-1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof)
  • Mcl-1 relate to a method for inhibiting the activity of Mcl-1 that can include contacting a cancer cell from a cancer described herein with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), and thereby inhibiting the activity of Mcl-1.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include inhibiting the activity of Mcl-1 using an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof).
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof).
  • inventions described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
  • Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
  • Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a cancer cell with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the compound inhibits the activity of Mcl-1.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • Some embodiments disclosed herein relate to a method for inhibiting the activity of Mcl-1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein or a cancer cell from a cancer described herein.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • inventions disclosed herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of Mcl-1.
  • Still other embodiments disclosed herein relate to a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of Mcl- 1.
  • suitable cancers include, but are not limited to: hematological malignancies (such as acute myeloid leukemia, multiple myeloma, mantle cell lymphoma, chronic lymphocytic leukemia, diffuse large B cell lymphoma, Burkitt’s lymphoma, follicular lymphoma) and solid tumors, for example, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), breast cancer, neuroblastoma, prostate cancer, melanoma, pancreatic cancer, uterine, endometrial, colon, oesophagus and liver cancers, osteosarcoma, Hodgkin lymphoma, mesothelioma, meningioma, glioma and tumors of upper aerodigestive, ovarian, thyroid, stomach and urinary tract.
  • NSCLC non-small cell lung cancer
  • SCLC small cell lung cancer
  • breast cancer neuroblastoma
  • prostate cancer prostate cancer
  • the cancer can be selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic
  • a cancer can become resistant to one or more anti-cancer agents.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can be used to treat and/or ameliorate a cancer that has become resistant to one or more anti-cancer agents (such as one or more Mcl-1 inhibitors).
  • anti-cancer agents examples include, but are not limited to, Mcl-1 inhibitors (such as AT101, gambogic acid, TW-37, AZD5991, Sabutoclax (BI-97C1), Maritoclax, UMI-77, A-1210477, S63845, MIK665/S64315, (-)BI97D6 and/or AMG176).
  • Mcl-1 inhibitors such as AT101, gambogic acid, TW-37, AZD5991, Sabutoclax (BI-97C1), Maritoclax, UMI-77, A-1210477, S63845, MIK665/S64315, (-)BI97D6 and/or AMG176.
  • the cancer that has become resistant to one or more anti-cancer agents can be a cancer described herein.
  • Mcl-1 inhibitors can cause one or more undesirable side effects in the subject being treated.
  • undesirable side effects include, but are not limited to, thrombocytopenia, neutropenia, anemia, diarrhea, vomiting, nausea, abdominal pain, and constipation.
  • a compound described herein for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof can result in a severity of a side effect (such as one of those described herein) that is 25% less than compared to the severity of the same side effect experienced by a subject receiving a known Mcl-1 inhibitor (such as AT101, gambogic acid, TW-37, AZD5991, Sabutoclax (BI-97C1), Maritoclax, UMI-77, A-1210477, S63845, MIK665/S64315, (-)BI97D6 and/or AMG176).
  • a side effect such as one of those described herein
  • a side effect such as one of those described herein
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof results in a number of side effects that is 25% less than compared to the number of side effects experienced by a subject receiving a known Mcl-1. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a severity of a side effect (such as one of those described herein) that is less in the range of about 10% to about 30% compared to the severity of the same side effect experienced by a subject receiving a known Mcl- 1.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof results in a number of side effects that is in the range of about 10% to about 30% less than compared to the number of side effects experienced by a subject receiving a known Mcl-1.
  • a “subject” refers to an animal that is the object of treatment, observation or experiment.
  • Animal includes cold- and warm-blooded vertebrates and invertebrates such as fish, shellfish, reptiles and, in particular, mammals.
  • “Mammal” includes, without limitation, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, and, in particular, humans.
  • the subject can be human.
  • the subject can be a child and/or an infant, for example, a child or infant with a fever.
  • the subject can be an adult.
  • treat do not necessarily mean total cure or abolition of the disease or condition. Any alleviation of any undesired signs or symptoms of the disease or condition, to any extent can be considered treatment and/or therapy. Furthermore, treatment may include acts that may worsen the subject’s overall feeling of well-being or appearance.
  • terapéuticaally effective amount and “effective amount” are used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated.
  • a therapeutically effective amount of compound, salt or composition can be the amount needed to prevent, alleviate or ameliorate symptoms of the disease or condition, or prolong the survival of the subject being treated. This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease or condition being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein.
  • the therapeutically effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated and the physical characteristics of the specific animal under consideration.
  • the dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.
  • an effective amount of a compound, or radiation is the amount that results in: (a) the reduction, alleviation or disappearance of one or more symptoms caused by the cancer, (b) the reduction of tumor size, (c) the elimination of the tumor, and/or (d) long-term disease stabilization (growth arrest) of the tumor.
  • a therapeutically effective amount is that amount that alleviates or eliminates cough, shortness of breath and/or pain.
  • an effective amount, or a therapeutically effective amount of a Mcl-1 inhibitor is the amount which results in the reduction in Mcl-1 activity and/or phosphorylation (such as phosphorylation of CDC2).
  • Mcl-1 activity is known to those skilled in the art and can be determined by the analysis of Mcl-1 intrinsic kinase activity and downstream substrate phosphorylation.
  • the amount of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature and/or symptoms of the disease or condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. In cases of administration of a pharmaceutically acceptable salt, dosages may be calculated as the free base. As will be understood by those of skill in the art, in certain situations it may be necessary to administer the compounds disclosed herein in amounts that exceed, or even far exceed, the dosage ranges described herein in order to effectively and aggressively treat particularly aggressive diseases or conditions.
  • a suitable dose will often be in the range of from about 0.05 mg/kg to about 10 mg/kg.
  • a suitable dose may be in the range from about 0.10 mg/kg to about 7.5 mg/kg of body weight per day, such as about 0.15 mg/kg to about 5.0 mg/kg of body weight of the recipient per day, about 0.2 mg/kg to 4.0 mg/kg of body weight of the recipient per day, or any amount in between.
  • the compound may be administered in unit dosage form; for example, containing 1 to 500 mg, 10 to 100 mg, 5 to 50 mg or any amount in between, of active ingredient per unit dosage form.
  • the desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day.
  • the sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.
  • the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight, the severity of the affliction, the mammalian species treated, the particular compounds employed and the specific use for which these compounds are employed.
  • the determination of effective dosage levels can be accomplished by one skilled in the art using routine methods, for example, human clinical trials, in vivo studies and in vitro studies.
  • useful dosages of a compound of Formula (I), or pharmaceutically acceptable salts thereof can be determined by comparing their in vitro activity, and in vivo activity in animal models.
  • Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the modulating effects, or minimal effective concentration (MEC).
  • MEC minimal effective concentration
  • the MEC will vary for each compound but can be estimated from in vivo and/or in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC value.
  • Compositions should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.
  • the attending physician would know how to and when to terminate, interrupt or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity).
  • the magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the disease or condition to be treated and to the route of administration. The severity of the disease or condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency, will also vary according to the age, body weight and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.
  • Compounds, salts and compositions disclosed herein can be evaluated for efficacy and toxicity using known methods.
  • the toxicology of a particular compound, or of a subset of the compounds, sharing certain chemical moieties may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of such studies are often predictive of toxicity in animals, such as mammals, or more specifically, humans.
  • the toxicity of particular compounds in an animal model such as mice, rats, rabbits, dogs or monkeys, may be determined using known methods.
  • the efficacy of a particular compound may be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials.
  • Example 1A (Ra)-(+)-(Z)-1 6 -Chloro-2 1 -ethyl-1 1 ,2 3 ,6 1 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0102] To a stirred solution of Intermediate 25A (200 mg, 0.285 mmol) in MeOH/THF/H2O (1:1:1, 6 mL) was added LiOH ⁇ H2O (180 mg, 4.29 mmol) at rt.
  • Example 1A (189 mg, 96%) as an off-white solid.
  • Example 1B (Sa)-(-)-(Z)-1 6 -Chloro-2 1 -ethyl-1 1 ,2 3 ,6 1 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0103] To a stirred 25B (200 mg, 0.285 mmol) in MeOH/THF/H2O (1:1:1, 6 mL) was added LiOH ⁇ H2O (180 mg, 4.29 mmol) at rt.
  • the reaction was stirred at 80 °C for 2 h. Upon completion, the solvent was evaporated. The aqueous layer was acidified to pH 6 using 2 N aq. HCl. The solid was filtered and washed with water (5 mL).
  • Example 1B The absolute stereochemistry of Example 1B was arbitrarily assigned.
  • Intermediate 26 (1,3-Dimethyl-1H-pyrazol-5-yl)methanol [0104] To a stirred solution imethyl-1H-pyrazole-5-carboxylate (25.0 g, 149 mmol) in THF (200 mL) was added 2.4 M LAH in THF (74.0 mL, 179 mmol) at 0°C. The reaction was stirred under N 2 for 3 h at 0 o C. The reaction was quenched with sat. Na 2 SO 4 (50 mL) at 0 o C. The white solid was filtered and washed with 10% MeOH in DCM (500 mL).
  • Example 2A (Ra)-(+)-(Z)-1 6 -Fluoro-1 1 ,2 1 ,2 3 ,6 1 -tetramethyl-1 1 H,2 1 H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0116]
  • Example 2A ermediate 37A following a similar procedure for the preparation of Example 1.
  • Example 2B (S a )-(-)-(Z)-1 6 -Fluoro-1 1 ,2 1 ,2 3 ,6 1 -tetramethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0117]
  • Example 2B ermediate 37B following a similar procedure for the preparation of Example 1.
  • Example 2B The absolute stereochemistry of Example 2B was arbitrarily assigned.
  • Intermediate 38 Methyl 7-bromo-3-(3-methoxy-3-oxopropyl)-6-methyl-1H-indole-2-carboxylate [0118]
  • Intermediate 38 m 2-bromo-3-methylaniline following a similar procedure for the preparation of Intermediate 1.
  • MS (LCMS) 354.2 [M+H] + .
  • Intermediate 39 Methyl 3-(3-acetoxypropyl)-7-bromo-1,6-dimethyl-1H-indole-2-carboxylate [0119]
  • Intermediate 39 was synthesized from Intermediate 38 following similar procedures for the preparation of Intermediates 2-4.
  • Example 3A (R a )-(+)-(Z)-1 1 ,1 6 ,2 1 ,2 3 ,6 1 -Pentamethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0126]
  • Example 3A ermediate 45A following a similar procedure for the preparation of Example 1.
  • Example 3B (S a )-(-)-(Z)-1 1 ,1 6 ,2 1 ,2 3 ,6 1 -Pentamethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0127]
  • Example 3B ermediate 45B following a similar procedure for the preparation of Example 1.
  • Example 3B The absolute stereochemistry of Example 3B was arbitrarily assigned.
  • Intermediate 46 Ethyl 5-(hydroxymethyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate [0128] To a stirred solution te 5 (30.0 g, 73.5 mmol) in THF (300 mL) was added NaHMDS (44.0 mL, 88.2 mmol) and NaI (5.50 g, 36.6 mmol) at 0 °C. The reaction was stirred for 30 min. PMBCl (13.85 g, 88.23 mmol) was added at 0 °C. The reaction was stirred at 50 °C for 2 h.
  • Example 4A (R a )-(+)-(Z)-1 6 -Chloro-1 1 ,2 1 ,2 3 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0145]
  • Example 4A ermediate 61A following a similar procedure for the preparation of Example 1.
  • Example 4B (S a )-(-)-(Z)-1 6 -Chloro-1 1 ,2 1 ,2 3 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0146]
  • Example 4B ermediate 61B following a similar procedure for the preparation of Example 1.
  • Example 4B The absolute stereochemistry of Example 4B was arbitrarily assigned.
  • Intermediate 62 Ethyl 4-oxo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate [0147] To a stirred solution H-pyrazole-3,5-dicarboxylate (20.0 g, 94.3 mmol) in DMF (200 mL) was added Cs 2 CO 3 (36.9 g, 113 mmol) and tert-butyl (2-bromoethyl) carbamate (26.29 g, 117.9 mmol) at 0 o C. The ice bath was removed, and the reaction was stirred at rt for 16 h.
  • the reaction Upon completion by TLC, the reaction was cooled to 0°C, diluted with EtOAc (500 mL) and diluted by the dropwise addition of water (29.2 mL). 15% NaOH (29.2 mL) and water (87.6 mL) was added, and the mixture was stirred at rt for 15 min. MgSO 4 was added, and the mixture was stirred for 15 min. The mixture was filtered through Celite, and the pad was washed with EtOAc (2 x 100 mL).
  • Example 5A (Ra)-(+)-(Z)-1 6 -Chloro-2 5 -ethyl-1 1 ,6 1 -dimethyl-2 4 ,2 5 ,2 6 ,2 7 -tetrahydro-1 1 H,6 1 H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0162]
  • Example 5 mediate 73A following a similar procedure for the preparation of Example 1.
  • Example 5A (190 mg, 70%), off-white solid; 98.6% chiral purity; 1H NMR (400 MHz, DMSO-d6) ⁇ 13.39 (br s, 1H), 11.8-11.0 (m, 1H), 8.17-8.08 (m, 1H), 7.95-7.80 (m, 1H), 7.78-7.70 (m, 1H), 7.54-7.43 (m, 2H), 7.38 (s, 1H), 7.20-1.08 (m, 1H), 6.63 (s, 1H), 4.90-4.73 (m, 1H), 4.60-3.80 (m, 9H), 3.75-3.40 (m, 9H), 3.30-2.80 (m, 6H), 2.40- 2.32 (m, 1H), 2.30-2.15 (m, 1H), 1.23 (brs, 3H); MS (LCMS) 727.4 [M+H] + .
  • Example 5A The absolute stereochemistry of Example 5A was arbitrarily assigned.
  • Example 5B (Sa)-(-)-(Z)-1 6 -Chloro-2 5 -ethyl-1 1 ,6 1 -dimethyl-2 4 ,2 5 ,2 6 ,2 7 -tetrahydro-1 1 H,6 1 H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0163]
  • Example 5 mediate 73B following a similar procedure for the preparation of Example 1.
  • Example 5B (212 mg, 77%), off-white solid; 99.2% chiral purity; 1H NMR (400 MHz, DMSO-d6) ⁇ 13.35 (br s, 1H), 11.50-10.91 (m, 1H), 8.15-8.08 (m, 1H), 7.92-7.80 (brs, 1H), 7.73-7.70 (m, 1H), 7.54-7.43 (m, 2H), 7.40 (s, 1H), 7.20-1.08 (m, 1H), 6.64 (s, 1H), 4.90-4.70 (m, 1H), 4.60-3.80 (m, 8H), 3.74 (s, 3H), 3.65-3.40 (m, 7H), 3.30- 2.80 (m, 6H), 2.40-2.32 (m, 1H), 2.30-2.15 (m, 1H), 1.23 (brs, 3H); MS (LCMS) 727.3 [M+H] + .
  • Example 5B The absolute stereochemistry of Example 5B was arbitrarily assigned.
  • Intermediate 74 Ethyl (Z)-5-((tert-butyldiphenylsilyl)oxy)-2-hydroxy-4-oxopent-2-enoate [0164] t-BuOK (3.60 kg d to THF (21 L), and the solution was cooled to 0 °C. Diethyl oxalate (4.69 kg, 32.1 mol) was added slowly, maintaining the temperature below 0 °C. The solution was stirred for 30 min at 0 °C. 1-((tert-Butyldiphenylsilyl)oxy)propan- 2-one (8.50 kg, 27.2 mol) was added slowly, maintaining the temperature below 0 °C.
  • Example 6A (Ra)-(+)-(Z)-1 6 -Chloro-2 1 -ethyl-9 6 -fluoro-1 1 ,2 3 ,6 1 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0181]
  • Example 6A mediate 87A following a similar procedure for the preparation of Example 1.
  • Example 6B (Sa)-(-)-(Z)-1 6 -Chloro-2 1 -ethyl-9 6 -fluoro-1 1 ,2 3 ,6 1 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0182]
  • Example 6B mediate 87B following a similar procedure for the preparation of Example 1.
  • Example 7A (Ra)-(+)-(Z)-1 6 ,9 6 -Difluoro-1 1 ,2 1 ,2 3 ,6 1 -tetramethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0185]
  • Example 7A rmediate 89A following a similar procedure for the preparation of Example 1.
  • Example 7B (S a )-(-)-(Z)-1 6 ,9 6 -Difluoro-1 1 ,2 1 ,2 3 ,6 1 -tetramethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0186]
  • Example 7B was synthesized from Intermediate 89B following a similar procedure for the preparation of Example 1.
  • Example 7B The absolute stereochemistry of Example 7B was arbitrarily assigned.
  • Intermediate 90 Methyl 7-(5-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H-pyrazol-3- yl)methyl)thio)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1,6-dimethyl-1H- indole-2-carboxylate OH following a similar procedure for the preparation of Intermediate 24.
  • Example 8A (Ra)-(+)-(Z)-9 6 -Fluoro-1 1 ,1 6 ,2 1 ,2 3 ,6 1 -pentamethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0189]
  • Example 8A rmediate 91A following a similar procedure for the preparation of Example 1.
  • Example 8B (Sa)-(-)-(Z)-9 6 -Fluoro-1 1 ,1 6 ,2 1 ,2 3 ,6 1 -pentamethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0190]
  • Example 8B rmediate 91B following a similar procedure for the preparation of Example 1.
  • Example 8B The absolute stereochemistry of Example 8B was arbitrarily assigned.
  • Intermediate 92 Methyl 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-2-carboxylate N N O O [0191] To a stirred solution dro-4H-pyrrolo[1,2-b]pyrazole-2-carboxylic acid (10.0 g, 65.7 mmol) in MeOH (100 mL) was added SOCl 2 (15.64 g, 131.4 mmol) at 0 °C. The reaction was stirred at reflux for 6 h. Upon completion by TLC, the solvent was evaporated and co-distilled with MeOH. The solid was dried under high vacuum to afford Intermediate 92 (10 g, 92%) as an off-white solid.
  • Example 9A (Ra)-(+)-(Z)-1 6 -Chloro-9 6 -fluoro-1 1 ,6 1 -dimethyl-2 5 ,2 6 -dihydro-1 1 H,2 4 H,6 1 H-10-oxa-4,8-dithia- 1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0202]
  • Example 9 mediate 102A following a similar procedure for the preparation of Example 1.
  • Example 9B (S a )-(-)-(Z)-1 6 -Chloro-9 6 -fluoro-1 1 ,6 1 -dimethyl-2 5 ,2 6 -dihydro-1 1 H,2 4 H,6 1 H-10-oxa-4,8-dithia- 1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0203]
  • Example 9B mediate 102B following a similar procedure for the preparation of Example 1.
  • Example 10A (Ra)-(+)-(Z)-1 6 -Chloro-9 6 -fluoro-1 1 ,6 1 -dimethyl-2 4 ,2 5 ,2 6 ,2 7 -tetrahydro-1 1 H,6 1 H-10-oxa-4,8- dithia-2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0211]
  • Example 10 mediate 109A following a similar procedure for the preparation of Example 1.
  • Example 10B (Sa)-(-)-(Z)-1 6 -Chloro-9 6 -fluoro-1 1 ,6 1 -dimethyl-2 4 ,2 5 ,2 6 ,2 7 -tetrahydro-1 1 H,6 1 H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0212]
  • Example 10 mediate 109B following a similar procedure for the preparation of Example 1.
  • Example 10B The absolute stereochemistry of Example 10B was arbitrarily assigned.
  • Intermediate 110 Ethyl 3-(((tert-butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazole-5-carboxylate [0213] To a stirred soluti 5 (47.0 g, 115 mmol) in DMF (450 mL) was added Cs2CO3 (112 g, 345 mmol) and MeI (14.3 mL, 230 mmol) at 0°C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by LCMS, water (800 mL) was added. The mixture was extracted with EtOAc (3 x 500 mL).
  • Example 11A (Ra)-(+)-(Z)-1 6 -Chloro-96-fluoro-1 1 ,2 1 ,2 3 ,6 1 -tetramethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0228]
  • Example 11 mediate 121A following a similar procedure for the preparation of Example 1.
  • Example 11A (70 mg, 60%), off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d 6 ) ⁇ 13.30 (br s, 1H), 8.27-8.22 (m, 1H), 7.58-7.52 (m, 2H), 7.38-7.32 (m, 2H), 6.83-6.78 (m, 2H), 4.79 (s, 1H), 4.08-3.90 (m, 4H), 3.80 (s, 3H), 3.62- 3.52 (m, 2H), 3.50 (s, 3H), 3.45-3.33 (m, 4H), 3.20-3.10 (m, 3H), 2.45-2.35 (m, 1H), 2.25-2.10 (m, 1H), 1.85 (s, 3H); MS (LCMS) 690.7 [M+H] + .
  • Example 11B (Sa)-(-)-(Z)-1 6 -Chloro-96-fluoro-1 1 ,2 1 ,2 3 ,6 1 -tetramethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0229]
  • Example 11 mediate 121B following a similar procedure for the preparation of Example 1.
  • Example 11B (70 mg, 60%), off-white solid; 97.4% chiral purity; 1H NMR (400 MHz, DMSO-d6) ⁇ 13.30 (br s, 1H), 8.27-8.22 (m, 1H), 7.58-7.52 (m, 2H), 7.38-7.32 (m, 2H), 6.84-6.78 (m, 2H), 4.79 (s, 1H), 4.08-3.90 (m, 4H), 3.80 (s, 3H), 3.62- 3.52 (m, 2H), 3.50 (s, 3H), 3.45-3.33 (m, 4H), 3.20-3.10 (m, 3H), 2.45-2.35 (m, 1H), 2.25-2.10 (m, 1H), 1.86 (s, 3H); MS (LCMS) 690.7 [M+H] + .
  • Example 11B The absolute stereochemistry of Example 11B was arbitrarily assigned.
  • Intermediate 122 Ethyl 5-((acetylthio)methyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate [0230] To a stirred solution te 47 (15.5 g, 50.2 mmol) in CH 3 CN (155 mL) was added KSAc (16.66 g, 100.4 mmol) and KI (5.72 g, 50.2 mmol) at 0 °C. The reaction was stirred at rt for 16 h. The reaction was quenched with water (100 mL) and extracted with EtOAc (3 x 100 mL).
  • the reaction was heated at 100°C for 3 h.
  • the reaction was cooled to rt, diluted with EtOAc (250 mL) and passed through a Celite pad. The pad was washed with EtOAc (100 mL). The filtrate was washed with brine (200 mL), dried (Na 2 SO 4 ), filtered and evaporated.
  • the crude residue was purified by flash chromatography (SiO 2 , 30% EtOAc/Pet.
  • Example 12A (R a )-(+)-(Z)-1 6 -Chloro-9 6 -fluoro-1 1 ,2 1 ,2 3 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid
  • Example 12B (Sa)-(-)-(Z)-1 6 -Chloro-9 6 -fluoro-1 1 ,2 1 ,2 3 -trimethyl-1 1 H,2 1 H,6 1 H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 -carboxylic acid [0237] To a stirred 128 (320
  • Example 12A and Example 12B The absolute stereochemistry of Example 12A and Example 12B was arbitrarily assigned.
  • Intermediate 129 Ethyl 5-(chloromethyl)-1-ethyl-1H-pyrazole-3-carboxylate [0238] To a stirred solut l 5-(chloromethyl)-1-ethyl-1H-pyrazole-3- carboxylate (5.00 g, 25.3 mmol) in DCM (50 mL) was added SOCl 2 (2.20 mL, 30.3 mmol) at 0 °C. The reaction was stirred at rt for 1 h. Upon completion by TLC, the reaction was diluted with DCM (50 mL) and washed with sat. NaHCO 3 (50 mL).
  • Example 13A (Ra)-(+)-(Z)-1 5 -Chloro-6 1 -ethyl-1 3 -methyl-2 5 ,2 6 -dihydro-1 1 H,2 4 H,6 1 H-10-oxa-4,8-dithia-1(4,1)- indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 - carboxylic acid [0252]
  • Example 13A rmediate 140A following a similar procedure for the preparation of Example 1.
  • Example 13B (Sa)-(-)-(Z)-1 5 -Chloro-6 1 -ethyl-1 3 -methyl-2 5 ,2 6 -dihydro-1 1 H,2 4 H,6 1 H-10-oxa-4,8-dithia-1(4,1)- indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)-naphthalenacyclotridecaphane-1 2 - carboxylic acid [0253]
  • Example 13B rmediate 140B following a similar procedure for the preparation of Example 1.
  • Example 13B The absolute stereochemistry of Example 13B was arbitrarily assigned.
  • Intermediate 141 Diethyl 1-isopropyl-1H-pyrazole-3,5-dicarboxylate [0254] To an ice cooled, st of diethyl 1H-pyrazole-3,5-dicarboxylate (50.0 g, 236 mmol) in DMF (250 mL) was added 60% NaH (10.3 g, 259 mmol) portionwise. The mixture was stirred at 0 o C for 30 min. Iodopropane (44.0 g, 259 mmol) was added slowly. The reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction was quenched with water (500 mL).
  • Example 14A (Ra)-(+)-(Z)-1 5 -Chloro-6 1 -isopropyl-1 3 -methyl-2 5 ,2 6 -dihydro-1 1 H,2 4 H,6 1 H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0262]
  • Example 14A rmediate 148A following a similar procedure for the preparation of Example 1.
  • Example 14B (Sa)-(-)-(Z)-1 5 -Chloro-6 1 -isopropyl-1 3 -methyl-2 5 ,2 6 -dihydro-1 1 H,2 4 H,6 1 H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-1 2 -carboxylic acid [0263]
  • Example 14B rmediate 148B following a similar procedure for the preparation of Example 1.
  • Example 14B The absolute stereochemistry of Example 14B was arbitrarily assigned.
  • HTRF Time Resolved Fluorescence
  • Assay conditions 4 nM Mcl-1, 100 nM FAM-Bak peptide, in 20 mM K Phosphate, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.005% Triton X-100 and 1% DMSO (final).
  • Assay procedure Compounds were tested in 10-dose IC50 mode, in singlicate, with 3-fold serial dilution starting at 10 ⁇ M or 1 ⁇ M. Compound stock solutions were added to protein solution using Acoustic technology. The compounds were then incubated with protein for 10 min at rt. The respective FAM labeled peptide was added and incubated for another 10 min. Anti-GST-Tb was added.
  • Example B NCI-H929 Cell Proliferation Assay [0265] Cell proliferation was measured using the CellTiter-Glo® Luminescent Cell Viability Assay. The assay involved the addition of a single reagent (CellTiter-Glo® Reagent) directly to cells cultured in serum-supplemented medium. NCI-H929 (ATCC CRL-9068) cells were cultured according to ATCC recommendations and were seeded at 3,000 cells per well.
  • Each compound evaluated was prepared as a DMSO stock solution (10 mM). Compounds were tested in duplicate on each plate, with a 10-point serial dilution curve (1:3 dilution). Compound treatment (1.0 ⁇ L) was added from the compound dilution plate to the cell plate. The highest compound concentration was 10 ⁇ M (final), with a 0.1% final DMSO concentration. Plates were then incubated at 37 °C, 5% CO2. After 72 h of compound treatment, cell plates were equilibrated at rt for approximately 30 mins. An equi-volume amount of CellTiter- Glo® Reagent (40 ⁇ L) was added to each well.
  • IC50 of each compound was calculated using GraphPad Prism by nonlinear regression analysis. IC 50 values are provided in Table 1. Table 1 Examples Mcl 1 IC50 (nM) H929 IC50 (nM) Examples Mcl-1 IC50 (nM) H929 IC50 (nM) gle .

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Abstract

Compounds of Formula (I) are provided herein. Such compounds, as well as pharmaceutically acceptable salts and compositions thereof, are useful for treating diseases or conditions, including conditions characterized by excessive cellular proliferation.

Description

MACROCYCLIC COMPOUNDS
INCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS
[0001] Any and all applications for which a foreign or domestic priority claim is identified, for example, in the Application Data Sheet or Request as filed with the present application, are hereby incorporated by reference under 37 CFR 1.57, and Rules 4.18 and 20.6, including U.S. Provisional Application No. 63/063,058, filed August 7, 2020.
Field
[0002] The present application relates to compounds that are Mcl-1 inhibitors and methods of using them to treat conditions characterized by excessive cellular proliferation, such as cancer.
Description
[0003] Mcl-1 (myeloid cell leukemia- 1) is a member of the Bcl-2 family of proteins. MCL-1 is widely expressed in human tissues and is primarily located in the mitochondria in cells. Upregulation of Mcl-1 occurs in different cancer types. Additionally, overexpression of Mcl-1 has been linked to drug resistance to several cancer therapies.
SUMMARY
[0004] Some embodiments provide a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
[0005] Some embodiments disclosed herein relate to a pharmaceutical composition that can include an effective amount of one or more of compounds of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.
[0006] Some embodiments described herein relate to a method for ameliorating and/or treating a cancer described herein that can include administering an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein.
[0007] Some embodiments described herein relate to a method for inhibiting replication of a malignant growth or a tumor that can include contacting the growth or the tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the malignant growth or tumor is due to a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
[0008] Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
[0009] Some embodiments described herein relate to a method for inhibiting the activity of Mcl-1 in a cell that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of Mcl-1. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of Mcl- 1.
[0010] Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include inhibiting the activity of Mcl-1 using an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof). Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1.
DETAILED DESCRIPTION
[0011] Myeloid Cell Leukemia 1 (Mcl-1) is an important anti- apop to tic member of the BCL-2 family of proteins and a master regulator of cell survival. Amplification of the MCL1 gene and/or overexpression of the Mcl-1 protein has been observed in multiple cancer types and is commonly implicated in tumor development. MCL1 is one of the most frequently amplified genes in human cancers. In many malignancies, Mcl-1 is a critical survival factor and it has been shown to mediate drug resistance to a variety of anti-cancer agents. Mcl-1 promotes cell survival by binding to pro-apoptotic proteins like Bim, Noxa, Bak, and Bax and neutralizing their deathinducing activities. Inhibition of Mcl-1 thereby releases these pro-apoptotic proteins, often leading to the induction of apoptosis in tumor cells dependent on Mcl-1 for survival. Therapeutically targeting Mcl-1 alone or in combination with other therapies, therefore, is a promising strategy to treat a multitude of malignancies and to overcome drug resistance in several human cancers. Definitions [0012] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. All patents, applications, published applications and other publications referenced herein are incorporated by reference in their entirety unless stated otherwise. In the event that there are a plurality of definitions for a term herein, those in this section prevail unless stated otherwise. [0013] As used herein, “Ca to Cb” in which “a” and “b” are integers refer to the number of carbon atoms in a group. The indicated group can contain from “a” to “b”, inclusive, carbon atoms. Thus, for example, a “C1 to C4 alkyl” group refers to all alkyl groups having from 1 to 4 carbons, that is, CH3-, CH3CH2-, CH3CH2CH2-, (CH3)2CH-, CH3CH2CH2CH2-, CH3CH2CH(CH3)- and (CH3)3C-. If no “a” and “b” are designated, the broadest range described in these definitions is to be assumed. [0014] As used herein, the term “alkyl” refers to a fully saturated aliphatic hydrocarbon group. The alkyl moiety may be branched or straight chain. Examples of branched alkyl groups include, but are not limited to, iso-propyl, sec-butyl, iso-butyl, t-butyl and the like. Examples of straight chain alkyl groups include, but are not limited to, methyl, ethyl, n-propyl and n-butyl. An alkyl group may be substituted or unsubstituted. [0015] The term “halogen atom” or “halogen” as used herein, means any one of the radio-stable atoms of column 7 of the Periodic Table of the Elements, such as, fluorine, chlorine, bromine and iodine. [0016] Where the number of substituents is not specified (e.g. haloalkyl), there may be one or more substituents present. For example, “haloalkyl” may include one or more of the same or different halogens. As another example, “C1-C3 alkoxyphenyl” may include one or more of the same or different alkoxy groups containing one, two or three atoms. [0017] As used herein, a radical indicates species with a single, unpaired electron such that the species containing the radical can be covalently bonded to another species. Hence, in this context, a radical is not necessarily a free radical. Rather, a radical indicates a specific portion of a larger molecule. The term “radical” can be used interchangeably with the term “group.” [0018] The term “pharmaceutically acceptable salt” refers to a salt of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. In some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts can be obtained by reacting a compound with inorganic acids such as hydrohalic acid (e.g., hydrochloric acid or hydrobromic acid), a sulfuric acid, a nitric acid and a phosphoric acid (such as 2,3-dihydroxypropyl dihydrogen phosphate). Pharmaceutical salts can also be obtained by reacting a compound with an organic acid such as aliphatic or aromatic carboxylic or sulfonic acids, for example formic, acetic, succinic, lactic, malic, tartaric, citric, ascorbic, nicotinic, methanesulfonic, ethanesulfonic, p- toluenesulfonic, trifluoro acetic, benzoic, salicylic, 2-oxopentanedioic or naphthalenesulfonic acid. Pharmaceutical salts can also be obtained by reacting a compound with a base to form a salt such as an ammonium salt, an alkali metal salt, such as a sodium, a potassium or a lithium salt, an alkaline earth metal salt, such as a calcium or a magnesium salt, a salt of a carbonate, a salt of a bicarbonate, a salt of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C1-C7 alkylamine, cyclohexylamine, triethanolamine, ethylenediamine and salts with amino acids such as arginine and lysine. For compounds of Formula (I), those skilled in the art understand that when a salt is formed by protonation of a nitrogen-based group (for example, NH2), the nitrogen-based group can be associated with a positive charge (for example, NH2 can become NH3+) and the positive charge can be balanced by a negatively charged counterion (such as Cl ).
[0019] It is understood that, in any compound described herein having one or more chiral centers, if an absolute stereochemistry is not expressly indicated, then each center may independently be of R-configuration or S -configuration or a mixture thereof. Thus, the compounds provided herein may be enantiomerically pure, enantiomerically enriched, racemic mixture, diastereomerically pure, diastereomerically enriched or a stereoisomeric mixture. In addition, it is understood that, in any compound described herein having one or more double bond(s) generating geometrical isomers that can be defined as E or Z, each double bond may independently be E or Z a mixture thereof. Likewise, it is understood that, in any compound described, all tautomeric forms are also intended to be included.
[0020] It is to be understood that where compounds disclosed herein have unfilled valencies, then the valencies are to be filled with hydrogens or isotopes thereof, e.g., hydrogen- 1 (protium) and hydrogen-2 (deuterium).
[0021] It is understood that the compounds described herein can be labeled isotopically. Substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo halflife or reduced dosage requirements. Each chemical element as represented in a compound structure may include any isotope of said element. For example, in a compound structure a hydrogen atom may be explicitly disclosed or understood to be present in the compound. At any position of the compound that a hydrogen atom may be present, the hydrogen atom can be any isotope of hydrogen, including but not limited to hydrogen- 1 (protium) and hydrogen-2 (deuterium). Thus, reference herein to a compound encompasses all potential isotopic forms unless the context clearly dictates otherwise.
[0022] It is understood that the methods and combinations described herein include crystalline forms (also known as polymorphs, which include the different crystal packing arrangements of the same elemental composition of a compound), amorphous phases, salts, solvates and hydrates. In some embodiments, the compounds described herein exist in solvated forms with pharmaceutically acceptable solvents such as water, ethanol or the like. In other embodiments, the compounds described herein exist in unsolvated form. Solvates contain either stoichiometric or non- stoichiometric amounts of a solvent, and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol or the like. Hydrates are formed when the solvent is water or alcoholates are formed when the solvent is alcohol. In addition, the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein.
[0023] Where a range of values is provided, it is understood that the upper and lower limit, and each intervening value between the upper and lower limit of the range is encompassed within the embodiments.
[0024] Terms and phrases used in this application, and variations thereof, especially in the appended claims, unless otherwise expressly stated, should be construed as open ended as opposed to limiting. As examples of the foregoing, the term ‘including’ should be read to mean ‘including, without limitation,’ ‘including but not limited to,’ or the like; the term ‘comprising’ as used herein is synonymous with ‘including,’ ‘containing,’ or ‘characterized by,’ and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps; the term ‘having’ should be interpreted as ‘having at least;’ the term ‘includes’ should be interpreted as ‘includes but is not limited to;’ the term ‘example’ is used to provide exemplary instances of the item in discussion, not an exhaustive or limiting list thereof; and use of terms like ‘preferably,’ ‘preferred,’ ‘desired,’ or ‘desirable,’ and words of similar meaning should not be understood as implying that certain features are critical, essential, or even important to the structure or function, but instead as merely intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment. In addition, the term “comprising” is to be interpreted synonymously with the phrases "having at least" or "including at least". When used in the context of a compound, composition or device, the term "comprising" means that the compound, composition or device includes at least the recited features or components, but may also include additional features or components.
[0025] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity. The indefinite article “a” or “an” does not exclude a plurality. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.
Compounds
[0026] Some embodiments disclosed herein relate to a compound of Formula (I), or a pharmaceutically acceptable salt thereof, having the structure:
Figure imgf000009_0001
wherein: R1 can be fluoro, chloro or methyl; R2 can be hydrogen or fluoro; Ring A can be n
Figure imgf000010_0003
alkyl; and Ring B can , wherein R6 and R7 can be independently hydrogen or an unsubs [0027] The
Figure imgf000010_0002
ormula (I) can be substituted with a halogen or an unsubstituted alkyl. In some embodiments, R1 can be fluoro, such that the indole can have er re
Figure imgf000010_0001
o- substituted. When the naphthyl ring is unsubstituted, R2 can be hydrogen. When the naphthyl ring is fluoro-substituted R2 can be fluoro. [0029] Having a fluoro on the naphthyl ring can be advantageous. Examples of advantages include, but are not limited to, the following: increased potency, greater stability and/or more efficient interaction with the Mcl-1 protein. By having a fluoro-substituted naphthyl, there may be increased hydrophobic interaction between the compound and Mcl-1 protein (for example, Valine 274, Leucine 290 and/or isoleucine 294), and thus, the potency of a compound described herein (including pharmaceutically acceptable salts thereof) can be increased. The fluoro on the naphthyl can act as an electron-withdrawing group and mitigate the electronic density of the sulfur and/or oxygen attached to the naphthyl which may lead to more efficient compound/Mcl- 1 protein hydrophobic interaction and/or more metabolic stability. The fluoro on the naphthyl may block potential metabolic soft spots of the compound, and result in more stable compounds.
[0030] As provided herein, Ring B can be a pyrazole ring. The pyrazole ring can be unsubstituted or alkyl-substituted. In some embodiments, Ring B can be
Figure imgf000011_0001
. In other embodiments, Ring B can
Figure imgf000011_0004
[0031] As described herein, the pyrazole ring can be unsubstituted, for example, then R6 is hydrogen and/or R7 is hydrogen. Also as described herein, the pyrazole ring can be alkylsubstituted. The pyrazole ring can be alkyl-substituted when R6 and/or R7 is an unsubstituted Ci- 4 alkyl. Examples of an unsubstituted Ci-4 alkyls include methyl, ethyl, n-propyl, iso-propyl, n- butyl, sec-butyl, iso-butyl and tert-butyl. Exemplary Ring B moieties include the following:
Figure imgf000011_0002
[0032] Ring A can be a monocyclic nitrogen-containing ring or a bicyclic nitrogencontaining ring. When Ring A is a monocyclic nitrogen-containing ring, Ring A can be a pyrazole ring, such
Figure imgf000011_0003
The substitution on Ring A can vary. Examples of R3 and R4 combinations for Ring A include the following: R3/R4 R3/R4 R3/R4 R3/R4 meth l/meth l meth l/eth l meth l/n-ro l meth l/iso- ro l
Figure imgf000012_0004
[0033] Ring A can also be a bicyclic nitrogen-containing ring. The ring fused to the pyrazole ring of Ring A can be a five-membered carbocyclyl or a six-membered carbocyclyl. In some embodiments, Ring A ca . In some embodiments, Ring A can be
Figure imgf000012_0001
s, e
Figure imgf000012_0002
embodiments, Ring A can be . Ring A can be fused to a six-membered
Figure imgf000012_0003
In
Figure imgf000013_0001
some embodiments, Ring A can be. When Rin
Figure imgf000013_0002
, R5 can be an unsubstituted C1-4 alkyl. Example of suitable are described herein. For e 5
Figure imgf000013_0003
xample, R can be methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl and tert-butyl. [0034] Examples of compounds of Formula (I), and pharmaceutically acceptable salts thereof, include the following:
Figure imgf000013_0004
Figure imgf000014_0001
Figure imgf000015_0001
Figure imgf000016_0001
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000019_0001
pharmaceutically acceptable salt of any of the foregoing.
[0035] Additional examples of compounds of Formula (I), and pharmaceutically acceptable salts thereof, include the following:
Figure imgf000019_0002
Figure imgf000020_0001
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000024_0001
Figure imgf000025_0001
Figure imgf000026_0001
Figure imgf000027_0001
Figure imgf000028_0001
Figure imgf000029_0001
Figure imgf000030_0001
Figure imgf000031_0001
pharmaceutically acceptable salt of any of the foregoing.
[0036] Examples of compounds that can be used in a method and/or use described herein, along with pharmaceutically acceptable salts thereof, include the following:
Figure imgf000031_0002
Figure imgf000032_0001
pharmaceutically acceptable salt of any of the foregoing.
[0037] In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, a compound, or a pharmaceutically acceptable salt thereof, where when R1
Figure imgf000032_0002
In some embodiments, a compound of Formula (I), or a pharmaceutically
Figure imgf000032_0005
acceptable salt thereof, a compound, or a pharmaceutically acceptable salt thereof, where when R1
Figure imgf000032_0003
Figure imgf000032_0004
. In some embodiments, R6 cannot be methyl. In some embodiments, R7 cannot be methyl. In some embodiments, R1 cannot be chloro. In some embodiments, R2 cannot be hydrogen. In some embodiments, R2 cannot be fluoro. In some embodiments, R1 cannot be chloro when R6 is methyl. In some embodiments, when R2 is hydrogen; then R1 cannot be chloro. In other embodiments, when R2 is fluoro; then R1 cannot be chloro. In some embodiments, when R2 is hydrogen; and R6 is methyl; then R1 cannot be chloro. In other embodiments, when R2 is fluoro; and R6 is methyl; then R1 cannot be chloro.
[0038] In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, cannot a compound disclosed in WO 2018/178226 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, cannot a compound disclosed in WO 2017/181625 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, cannot a compound disclosed in WO 2020/103864 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, cannot a compound disclosed in WO 2020/151738 that would be encompassed by a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
Synthesis
[0039] Compounds of the Formula (I), or pharmaceutically acceptable salts thereof, can be made in various ways by those skilled using known techniques as guided by the detailed teachings provided herein. For example, in an embodiment, compounds of the Formula (I) are prepared in accordance with General Scheme 1 as shown herein.
Figure imgf000033_0001
[0040] Compounds of Formula (I), and pharmaceutically acceptable salts thereof, can be prepared according to the preparation shown in Scheme 1. Compound A can undergo a Mitsunobu reaction and close the ring to form the macrocyclic Compound B. In Scheme 1, P represents a suitable protecting group. Removal of the protecting group via a hydrolysis reaction provides a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
Pharmaceutical Compositions
[0041] Some embodiments described herein relate to a pharmaceutical composition, that can include an effective amount of one or more compounds described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.
[0042] The term “pharmaceutical composition” refers to a mixture of one or more compounds and/or salts disclosed herein with other chemical components, such as diluents or carriers. The pharmaceutical composition facilitates administration of the compound to an organism. Pharmaceutical compositions can also be obtained by reacting compounds with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutical compositions will generally be tailored to the specific intended route of administration.
[0043] The term “physiologically acceptable” defines a carrier, diluent or excipient that does not abrogate the biological activity and properties of the compound nor cause appreciable damage or injury to an animal to which delivery of the composition is intended.
[0044] As used herein, a “carrier” refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, without limitation, dimethyl sulfoxide (DMSO) is a commonly utilized carrier that facilitates the uptake of many organic compounds into cells or tissues of a subject.
[0045] As used herein, a “diluent” refers to an ingredient in a pharmaceutical composition that lacks appreciable pharmacological activity but may be pharmaceutically necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and/or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the pH and isotonicity of human blood. [0046] As used herein, an “excipient” refers to an essentially inert substance that is added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the composition. For example, stabilizers such as anti-oxidants and metal-chelating agents are excipients. In an embodiment, the pharmaceutical composition comprises an anti-oxidant and/or a metal-chelating agent. A “diluent” is a type of excipient. [0047] The pharmaceutical compositions described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other active ingredients, as in combination therapy, or carriers, diluents, excipients or combinations thereof. Proper formulation is dependent upon the route of administration chosen. Techniques for formulation and administration of the compounds described herein are known to those skilled in the art. [0048] The pharmaceutical compositions disclosed herein may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes. Additionally, the active ingredients are contained in an amount effective to achieve its intended purpose. Many of the compounds used in the pharmaceutical combinations disclosed herein may be provided as salts with pharmaceutically compatible counterions. [0049] Multiple techniques of administering a compound, salt and/or composition exist in the art including, but not limited to, oral, rectal, pulmonary, topical, aerosol, injection, infusion and parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intranasal and intraocular injections. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, can be administered orally. [0050] One may also administer the compound, salt and/or composition in a local rather than systemic manner, for example, via injection or implantation of the compound directly into the affected area, often in a depot or sustained release formulation. Furthermore, one may administer the compound in a targeted drug delivery system, for example, in a liposome coated with a tissue-specific antibody. The liposomes will be targeted to and taken up selectively by the organ. For example, intranasal or pulmonary delivery to target a respiratory disease or condition may be desirable.
[0051] The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions that can include a compound and/or salt described herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
Uses and Methods of Treatment
[0052] Some embodiments described herein relate to a method for ameliorating and/or treating a cancer described herein that can include administering an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating and/or treating a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating and/or treating a cancer described herein. [0053] Some embodiments described herein relate to a method for inhibiting replication of a malignant growth or a tumor that can include contacting the growth or the tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the malignant growth or tumor is due to a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
[0054] Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a malignant growth or a tumor with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer that can include contacting a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer described herein.
[0055] Some embodiments described herein relate to a method for inhibiting the activity of Mcl-1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein. Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of Mcl-1. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of Mcl-1. Some embodiments described herein relate to a method for inhibiting the activity of Mcl-1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a cancer cell from a cancer described herein. Other embodiments described herein relate to a method for inhibiting the activity of Mcl-1 that can include contacting a cancer cell from a cancer described herein with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), and thereby inhibiting the activity of Mcl-1. [0056] Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include inhibiting the activity of Mcl-1 using an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof). Other embodiments described herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1. Still other embodiments described herein relate to an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for ameliorating or treating a cancer described herein by inhibiting the activity of Mcl-1. Some embodiments described herein relate to a method for ameliorating or treating a cancer described herein that can include contacting a cancer cell with an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof), wherein the compound inhibits the activity of Mcl-1.
[0057] Some embodiments disclosed herein relate to a method for inhibiting the activity of Mcl-1 that can include providing an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) to a subject having a cancer described herein or a cancer cell from a cancer described herein. Other embodiments disclosed herein relate to the use of an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) in the manufacture of a medicament for inhibiting the activity of Mcl-1. Still other embodiments disclosed herein relate to a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) for inhibiting the activity of Mcl- 1.
[0058] Examples of suitable cancers include, but are not limited to: hematological malignancies (such as acute myeloid leukemia, multiple myeloma, mantle cell lymphoma, chronic lymphocytic leukemia, diffuse large B cell lymphoma, Burkitt’s lymphoma, follicular lymphoma) and solid tumors, for example, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), breast cancer, neuroblastoma, prostate cancer, melanoma, pancreatic cancer, uterine, endometrial, colon, oesophagus and liver cancers, osteosarcoma, Hodgkin lymphoma, mesothelioma, meningioma, glioma and tumors of upper aerodigestive, ovarian, thyroid, stomach and urinary tract. In some embodiments, the cancer can be selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma.
[0059] As described herein, a cancer can become resistant to one or more anti-cancer agents. In some embodiments, a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition that includes an effective amount of a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can be used to treat and/or ameliorate a cancer that has become resistant to one or more anti-cancer agents (such as one or more Mcl-1 inhibitors). Examples of anti-cancer agents that a subject may have developed resistance to include, but are not limited to, Mcl-1 inhibitors (such as AT101, gambogic acid, TW-37, AZD5991, Sabutoclax (BI-97C1), Maritoclax, UMI-77, A-1210477, S63845, MIK665/S64315, (-)BI97D6 and/or AMG176). In some embodiments, the cancer that has become resistant to one or more anti-cancer agents can be a cancer described herein.
[0060] Several known Mcl-1 inhibitors can cause one or more undesirable side effects in the subject being treated. Examples of undesirable side effects include, but are not limited to, thrombocytopenia, neutropenia, anemia, diarrhea, vomiting, nausea, abdominal pain, and constipation. In some embodiments, a compound described herein (for example, a compound of Formula (I), or a pharmaceutically acceptable salt thereof) can decrease the number and/or severity of one or more side effects associated with a known Mcl-1 inhibitor. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, can result in a severity of a side effect (such as one of those described herein) that is 25% less than compared to the severity of the same side effect experienced by a subject receiving aknown Mcl-1 inhibitor (such as AT101, gambogic acid, TW-37, AZD5991, Sabutoclax (BI-97C1), Maritoclax, UMI-77, A-1210477, S63845, MIK665/S64315, (-)BI97D6 and/or AMG176). In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a number of side effects that is 25% less than compared to the number of side effects experienced by a subject receiving a known Mcl-1. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a severity of a side effect (such as one of those described herein) that is less in the range of about 10% to about 30% compared to the severity of the same side effect experienced by a subject receiving a known Mcl- 1. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, results in a number of side effects that is in the range of about 10% to about 30% less than compared to the number of side effects experienced by a subject receiving a known Mcl-1.
[0061] The one or more compounds of Formula (I), or a pharmaceutically acceptable salt thereof, that can be used to treat, ameliorate and/or inhibit the growth of a cancer wherein inhibiting the activity of Mcl-1 is beneficial is provided in any of the embodiments described in paragraphs [0064]-[0084], under the heading titled “Compounds.”
[0062] As used herein, a “subject” refers to an animal that is the object of treatment, observation or experiment. “Animal” includes cold- and warm-blooded vertebrates and invertebrates such as fish, shellfish, reptiles and, in particular, mammals. “Mammal” includes, without limitation, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, and, in particular, humans. In some embodiments, the subject can be human. In some embodiments, the subject can be a child and/or an infant, for example, a child or infant with a fever. In other embodiments, the subject can be an adult.
[0063] As used herein, the terms “treat,” “treating,” “treatment,” “therapeutic,” and “therapy” do not necessarily mean total cure or abolition of the disease or condition. Any alleviation of any undesired signs or symptoms of the disease or condition, to any extent can be considered treatment and/or therapy. Furthermore, treatment may include acts that may worsen the subject’s overall feeling of well-being or appearance.
[0064] The terms “therapeutically effective amount” and “effective amount” are used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated. For example, a therapeutically effective amount of compound, salt or composition can be the amount needed to prevent, alleviate or ameliorate symptoms of the disease or condition, or prolong the survival of the subject being treated. This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease or condition being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein. The therapeutically effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.
[0065] For example, an effective amount of a compound, or radiation, is the amount that results in: (a) the reduction, alleviation or disappearance of one or more symptoms caused by the cancer, (b) the reduction of tumor size, (c) the elimination of the tumor, and/or (d) long-term disease stabilization (growth arrest) of the tumor. In the treatment of lung cancer (such as nonsmall cell lung cancer) a therapeutically effective amount is that amount that alleviates or eliminates cough, shortness of breath and/or pain. As another example, an effective amount, or a therapeutically effective amount of a Mcl-1 inhibitor is the amount which results in the reduction in Mcl-1 activity and/or phosphorylation (such as phosphorylation of CDC2). The reduction in Mcl-1 activity is known to those skilled in the art and can be determined by the analysis of Mcl-1 intrinsic kinase activity and downstream substrate phosphorylation. [0066] The amount of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature and/or symptoms of the disease or condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. In cases of administration of a pharmaceutically acceptable salt, dosages may be calculated as the free base. As will be understood by those of skill in the art, in certain situations it may be necessary to administer the compounds disclosed herein in amounts that exceed, or even far exceed, the dosage ranges described herein in order to effectively and aggressively treat particularly aggressive diseases or conditions.
[0067] In general, however, a suitable dose will often be in the range of from about 0.05 mg/kg to about 10 mg/kg. For example, a suitable dose may be in the range from about 0.10 mg/kg to about 7.5 mg/kg of body weight per day, such as about 0.15 mg/kg to about 5.0 mg/kg of body weight of the recipient per day, about 0.2 mg/kg to 4.0 mg/kg of body weight of the recipient per day, or any amount in between. The compound may be administered in unit dosage form; for example, containing 1 to 500 mg, 10 to 100 mg, 5 to 50 mg or any amount in between, of active ingredient per unit dosage form.
[0068] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.
[0069] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight, the severity of the affliction, the mammalian species treated, the particular compounds employed and the specific use for which these compounds are employed. The determination of effective dosage levels, that is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine methods, for example, human clinical trials, in vivo studies and in vitro studies. For example, useful dosages of a compound of Formula (I), or pharmaceutically acceptable salts thereof, can be determined by comparing their in vitro activity, and in vivo activity in animal models. Such comparison can be done by comparison against an established drug, such as cisplatin and/or gemcitabine) [0070] Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the modulating effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vivo and/or in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC value. Compositions should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.
[0071] It should be noted that the attending physician would know how to and when to terminate, interrupt or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the disease or condition to be treated and to the route of administration. The severity of the disease or condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency, will also vary according to the age, body weight and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.
[0072] Compounds, salts and compositions disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of a particular compound, or of a subset of the compounds, sharing certain chemical moieties, may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of such studies are often predictive of toxicity in animals, such as mammals, or more specifically, humans. Alternatively, the toxicity of particular compounds in an animal model, such as mice, rats, rabbits, dogs or monkeys, may be determined using known methods. The efficacy of a particular compound may be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the art to choose an appropriate model, dose, route of administration and/or regime. EXAMPLES [0073] Additional embodiments are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the claims. Intermediate 1 Methyl 7-bromo-6-chloro-3-(3-methoxy-3-oxopropyl)-1H-indole-2-carboxylate [0074] To a stirred, 0 °C
Figure imgf000045_0001
-3-chloroaniline (25.0 g, 121 mmol) in conc. HCl (62.5 mL) and water (62.5 mL) was added a solution of NaNO2 (8.79 g, 127 mmol) in water (30 mL). The ice bath was removed, and the reaction was stirred at rt for 1.5 h. A solution of KOAc (167 g, 1.70 mol) in water (250 mL) was added, and the reaction was cooled to 0 oC. Methyl 2-oxocyclopentane-1-carboxylate (17.29 g, 121.3 mmol) was added dropwise, and the reaction was stirred at 0-5 °C for 30 min. The ice bath was removed, and the reaction was stirred at rt for 2 h. The solution was extracted with DCM (3 x 400 mL). The combined organic layers were washed with brine (200 mL), dried (Na2SO4) and filtered. The solvent evaporated to afford methyl 1-((2-bromo-3-chlorophenyl)diazenyl)-2-oxocyclopentane-1-carboxylate (42 g, 96%) as a red solid. MS (LCMS) 361.1 [M+H]+. [0075] To a stirred solution of methyl 1-((2-bromo-3-chlorophenyl)diazenyl)-2- oxocyclopentane-1-carboxylate (42.0 g, 117 mmol) in MeOH (420 mL) was added conc. H2SO4 (30.0 mL, 567 mol) at 0 °C. The reaction was stirred at 80 °C for 2 h. The mixture was cooled to room temperature (rt), and the solids were filtered and washed with MeOH to afford dimethyl (E/Z)-2-(2-(2-bromo-3-chlorophenyl)hydrazineylidene)hexanedioate (28 g, 61%) as a yellow solid. MS (LCMS) 393.2 [M+H]+. [0076] To a stirred solution of dimethyl (E/Z)-2-(2-(2-bromo-3- chlorophenyl)hydrazineylidene)-hexanedioate (29.0 g, 74.1 mmol) in MeOH (290 mL) was added conc. H2SO4 (50.0 mL, 938 mmol) at 0 °C. The reaction was stirred at 80 °C for 4 days. The reaction was cooled to rt, and the solid was filtered and washed with MeOH. The precipitate was dried under high vacuum to give Intermediate 1 (14 g, 50% yield) as an off-white solid. 1H NMR (400 MHz, CDCl3) δ 8.82 (br s, 1H), 7.61 (d, J=8.8 Hz, 1H), 7.23 (d, J=8.8 Hz, 1H), 3.98 (s, 3H), 3.63 (s, 3H), 3.37 (t, J=8.0 Hz, 2H), 2.68 (t, J=8.0 Hz, 2H); MS (LCMS) 375.9 [M+H]+. Intermediate 2 Methyl 7-bromo-6-chloro-3-(3-methoxy-3-oxopropyl)-1-methyl-1H-indole-2-carboxylate [0077] To a stirred soluti
Figure imgf000046_0001
1 (125 g, 373 mmol) in DMF (1.2 L) was added Cs2CO3 (65.3 g, 502 mmol) followed by MeI (95.14 g, 670.0 mmol) at 0 °C. The reaction was stirred at rt for 3 h. After completion, the reaction was quenched with ice water (1 L) and allowed to stir for 30 min where a solid precipitated. The solid was filtered, washed with n-pentane and dried under high vacuum to afford Intermediate 2 (90 g, 70%) as a brown solid. MS (LCMS) 388.0 [M+H]+. Intermediate 3 Methyl 7-bromo-6-chloro-3-(3-hydroxypropyl)-1-methyl-1H-indole-2-carboxylate [0078] To a stirred, 0 °C
Figure imgf000046_0002
ediate 2 (125 g, 322 mmol) in THF (1.2 L) was added 1 M BH3 .THF in THF (1.77 L) over 30 min. The ice bath was removed, and the reaction was stirred at rt for 4 h. Upon completion by TLC, the reaction was cooled to 0 oC and quenched with methanol (1770 mL) and 6 N HCl (1770 mL). The mixture was extracted with EtOAc (2 x 1 L). The combined organic layers were washed with brine (1 L) and dried (Na2SO4). The solvent removed under reduced pressure to afford Intermediate 3 (130 g) as a brown solid. MS (LCMS) 362.0 [M+H]+. Intermediate 4 Methyl 3-(3-acetoxypropyl)-7-bromo-6-chloro-1-methyl-1H-indole-2-carboxylate
Figure imgf000046_0003
[0079] To a stirred, 0 °C solution of Intermediate 3 (125 g, 322 mmol) in DCM (1.2 L) was added Et3N (70.66 g, 698.0 mmol) and DMAP (3 g) followed by Ac2O (53.4 g, 524 mmol). The ice bath was removed, and the reaction was stirred at rt for 1 h. Upon completion, the reaction was diluted with water (1 L) at 0 °C and extracted with DCM (2 x 1 L). The combined organic layers were washed with brine (1 L) and dried (Na2SO4). The solvent was evaporated, and the residue was purified by flash chromatography (SiO2, EtOAc) to afford Intermediate 4 (96.3 g, 74%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.50-7.47 (d, J=12.0 Hz, 1H), 7.24-7.21 (d, J=11.2 Hz, 1H), 4.32 (s, 3H), 4.08 (t, J=8.8 Hz 2H), 3.95 (s, 3H), 3.04 (t, J=10.4 Hz, 2H), 2.07 (s, 3H), 1.96 (m, 2H); MS (LCMS) 404.3 [M+H]+. Intermediate 5 Ethyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-1H-pyrazole-3-carboxylate [0080] To a stirred soluti
Figure imgf000047_0001
rt-butyldiphenylsilyl)oxy)-2-hydroxy-4- oxopent-2-enoate (250 g, 607 mmol) in EtOH (250 mL) was added 90-99% of N2H2.H2O (33.6 mL, 607 mmol). The reaction was stirred at rt 30 min. The reaction was concentrated, diluted with cold water (1 L) and extracted with EtOAc (2 x 1 L). The combined organic layers were washed with brine (1.5 L), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 15% EtOAc/Pet. ether) to afford Intermediate 5 (49 g, 20%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.72-7.62 (m, 4H), 7.45-7.35 (m, 6H), 6.67 (s, 1H), 4.79 (s, 2H), 4.41-4.33 (m, 2H), 1.41-1.36 (m, 3H), 1.08 (s, 9H). Intermediate 6 Ethyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazole-3-carboxylate [0081] To a stirred solutio
Figure imgf000047_0002
5 (110 g, 270 mmol) in THF (1.1 L) was added 1M NaHMDS in THF (323.5 mL, 323.5 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 30 min. Methyl iodide (20.32 mL, 323.5 mmol) was added, and the reaction was stirred for 2 h at rt. The reaction was quenched with sat. NH4Cl (1 L) and extracted with EtOAc (1 L). The organic layer was separated, washed with brine (1 L), dried (Na2SO4), filtered and evaporated to afford a mixture of 5-(((tert-Butyldiphenylsilyl)oxy)methyl)-1-methyl- 1H-pyrazole-3-carboxylic acid and Intermediate 6 (130 g) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.73-7.61 (m, 4H), 7.45-7.35 (m, 6H), 6.55 (s, 1H), 4.66 (s, 2H), 4.39 (q, J=6.8 Hz, 2H), 3.93 (s, 3H), 1.39 (t, J=7.1 Hz, 3H), 1.07 (s, 9H). MS (LCMS) 423.3 [M+H]+. Intermediate 7 (5-(((tert-Butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazol-3-yl)methanol [0082] To a stirred soluti
Figure imgf000048_0001
6 (130 g, 307 mmol) in THF (650 mL) was added 2.4 M LAH in THF (128 mL, 307 mmol) at 0 °C. The reaction was stirred at 0 oC for 1 h. Upon completion by TLC, the reaction was quenched with sat. Na2SO4. The mixture was filtered through a Celite pad. The filtrate was dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 7 (45 g, 44%, 2-steps) as an oil. MS (LCMS) 381.3 [M+H]+. Intermediate 8 (5-(((tert-Butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazol-3-yl)methyl acetate [0083] To a stirred soluti
Figure imgf000048_0002
7 (230 g, 605 mmol) in DCM (2.3 L) was added Et3N (170 mL, 1.21 mol), DMAP (5 g), and Ac2O (68.0 mL, 726 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 6 h. Upon completion by TLC, the reaction was diluted with water (2 L) and extracted with DCM (2 x 1 L). The combined organic layers were washed with brine (1 L), dried (Na2SO4), filtered and concentrated to afford Intermediate 8 (220g) as an oil. MS (LCMS) 423.2 [M+H]+. Intermediate 9 (5-(Hydroxymethyl)-1-methyl-1H-pyrazol-3-yl)methyl acetate [0084] To a stirred solution
Figure imgf000048_0003
e 8 (110 g, 261 mmol) in THF (1.1 L) was added CsF (78.0 g, 521 mmol) and 1M TBAF in THF (104 mL, 104 mmol). The reaction was stirred at rt for 4 h. Upon completion by TLC, the reaction was filtered, and the solvent was removed under reduced pressure. The crude material was purified by flash chromatography (SiO2, EtOAc) to afford Intermediate 9 (62 g) as an off-white solid. MS (LCMS) 185.1 [M+H]+. Intermediate 10 (5-(Chloromethyl)-1-methyl-1H-pyrazol-3-yl)methyl acetate [0085] To a stirred solution
Figure imgf000049_0001
te 9 (16.0 g, 86.9 mmol) in DCM (160 mL) under argon was added SOCl2 (12.0 mL, 174 mmol) at 0 °C. The reaction was stirred at rt for 3 h. The volatiles were evaporated, and the crude residue was quenched with sat. NaHCO3 (150 mL). The mixture was extracted with DCM (2 x 150 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated to afford Intermediate 10 (18 g) as a brown solid. MS (LCMS) 203.1 [M+H]+. Intermediate 11 3-(Acetylthio)naphthalen-1-yl acetate [0086] To a stirred solution
Figure imgf000049_0002
ydroxynaphthalene-2-sulfonate (100 g, 406 mmol), TPP (383 g, 1.46 mol) in CH3CN (1 L) was added I2 (82.5 g, 325 mmol) at rt. The reaction was stirred at 85 °C for 12 h. The reaction was cooled to 0 °C and stirred for 2 h. The waste solids were removed by filtration and washed with CH3CN (2 x 250 mL). The filtrate was concentrated and charged into a reactor. [0087] To a stirred solution of 3-mercaptonaphthalen-1-ol (406 mmol) in CH3CN (1.5 L) was added Et3N (166 mL, 1.22 mmol), DMAP (5.0 g), and Ac2O (76.9 mL, 813 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 2 h. Upon completion by TLC, the solvent was removed, and the mixture was diluted with water (1 L). The mixture was extracted with DCM (2 x 1 L). The combined organic layers were washed with brine (1 L), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 10% EtOAc/Pet. ether) to afford Intermediate 11 (78 g, 75%, 2-steps) as a yellow solid. MS (LCMS) 261.0 [M+H]+. Intermediate 12 3-(((3-(Hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-ol [0088] To a stirred, d
Figure imgf000050_0001
mediate 10 (18.0 g, 89.1 mmol) in MeOH (90 mL) and THF (36 mL), was added K2CO3 (61.5 g, 446 mmol) and Intermediate 11 (23.2 g, 89.1 mmol). The reaction was stirred at rt for 4 h. The reaction was filtered, and the filtrate was concentrated. The residue was triturated with a mixture of DCM (50 mL) and n- pentane (50 mL). The solid was collected by filtration and dried under high vacuum to afford Intermediate 12 (12 g, 45%) as a brown solid. MS (LCMS) 301.1 [M+H]+. Intermediate 13 3-(((3-(Chloromethyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-ol [0089] To a stirred solu
Figure imgf000050_0002
(40.0 g, 66.6 mmol) in DMF (200 mL) under argon was added MsCl (7.70 mL, 99.9 mmol) and LiCl (4.19 g, 99.9 mmol) at 0 oC. The ice bath was removed, and the reaction was stirred at rt for 3 h. Upon completion by TLC, the reaction was quenched with cold water (400 mL) and then extracted with EtOAc (2 x 400 mL). The combined organic layers were washed with brine (500 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 13 (35.0 g) as a brown liquid. MS (LCMS) 319.2 [M+H]+. Intermediate 14 3-(((3-(Chloromethyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-yl acetate [0090] To a stirred solu
Figure imgf000050_0003
(66.6 mmol) in CH3CN (350 mL) was added DMAP (1.34 g, 11.0 mmol) and Ac2O (16.8 mL, 164 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 3 h. Upon completion, the reaction was concentrated, and water (500 mL) was added. The mixture was extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (1 L), dried (Na2SO4), filtered and evaporated to afford Intermediate 14 (32.2 g) as a brown solid. MS (LCMS) 361.2 [M+H]+. Intermediate 15 3-(((3-((Acetylthio)methyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-yl acetate [0091] To a stirred sol
Figure imgf000051_0001
32.0 g, 88.6 mmol) in ACN (320 mL) was added KSAc (15.15 g, 133.0 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 3 h. Upon completion, the reaction was concentrated and water (300 mL) was added. The mixture was extracted with EtOAc (2 x 300 mL). The combined organic layers were washed with brine (500 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford Intermediate 15 (28 g, 53%, 3-steps) as a yellow solid. MS (LCMS) 401.2 [M+H]+. Intermediate 16 (1-Ethyl-3-methyl-1H-pyrazol-5-yl)methanol [0092] To a stirred solution
Figure imgf000051_0002
methyl-1H-pyrazole-5-carboxylic acid (35.0 g, 227 mmol) in THF (350 mL) was added 2.4M LiAlH4 in THF (104 mL, 249 mmol) at 0 °C. The reaction was stirred at rt for 2 h. Upon completion by LCMS, the reaction was quenched with cold sat. Na2SO4 (200 mL) at 0°C. The reaction was filtered through a Celite bed and washed with EtOAc (2 x 250 mL). The filtrate was washed with brine (500 mL), dried (Na2SO4), filtered and concentrated under reduced pressure to afford Intermediate 16 (30 g, 94%) as an oil. MS (ESI) 141.1 [M+H]+. Intermediate 17 (4-Bromo-1-ethyl-3-methyl-1H-pyrazol-5-yl)methanol [0093] To a stirred solution o
Figure imgf000051_0003
ate 16 (30.0 g, 214 mmol) in DCM (300 mL) was added NBS (40.0 g, 225 mmol) portion wise over 30 min. at 0 °C. The ice bath was removed, and the reaction was stirred for 1 h at rt. Upon completion by LCMS, the reaction was quenched with water (500 mL) and diluted with DCM (500 mL). The layers were separated, and the organic layer was washed with brine (500 mL), dried (Na2SO4), filtered and concentrated. The crude mixture was triturated with a mixture of n-pentane (300 mL) and diethyl ether (100 mL). The solid was filtered and dried under reduced pressure to afford Intermediate 17 (30 g, 64%) as an off-white solid. MS (ESI) 219.2 [M+H]+. Intermediate 18 4-Bromo-1-ethyl-5-(((4-methoxybenzyl)oxy)methyl)-3-methyl-1H-pyrazole [0094] To a stirred solution
Figure imgf000052_0001
e 17 (30.0 g, 137 mmol) in DMF (300 mL) was added 60 % NaH (8.20 g, 342 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 30 min. 1-(Chloromethyl)-4-methoxybenzene (32.0 g, 205.1 mmol) and KI (4.40 g, 26.5 mmol) was added, and the reaction was stirred at rt for 16 h. Upon completion, the reaction was quenched with ice, diluted with water (700 mL) and extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with water (2 x 500 mL) and brine (500 mL). The organic layer was dried (Na2SO4), filtered and concentrated. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 18 (28 g, 60%) as an oil. MS (LCMS) 339.2 [M+H]+. Intermediate 19 1-Ethyl-5-(((4-methoxybenzyl)oxy)methyl)-3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 2-yl)-1H-pyrazole [0095] To a stirred solution
Figure imgf000052_0002
te 18 (28.0 g, 82.8 mmol) in THF (300 mL) was added 1.6 M n-BuLi in hexanes (38.0 mL, 99.4 mmol) at -78 °C. The reaction was stirred for 1 h at -78 °C. 2-Isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (44.28 g, 238.1 mmol) was added at -78 °C, and the reaction was stirred at -78 oC for 1 h. Upon completion, the reaction was quenched with EtOAc (50 mL) and allowed to warm to rt. The solvent was removed, and the crude was diluted with EtOAc (300 mL). The mixture was filtered through a Celite pad which was washed with EtOAc (2 x 100 mL). The filtrate was evaporated, and the crude product was triturated with n-pentane. The solid was collected by filtration and dried under vacuum to afford Intermediate 19 (17 g, 53%) as an off-white solid. MS (ESI) 387.0 [M+H]+. Intermediate 20 Methyl 3-(3-acetoxypropyl)-6-chloro-7-(1-ethyl-5-(((4-methoxybenzyl)oxy)methyl)-3-methyl- 1H-pyrazol-4-yl)-1-methyl-1H-indole-2-carboxylate [0096] To a stirred soluti
Figure imgf000053_0001
19 (17.0 g, 44.0 mmol) in 1,4-dioxane (170 mL) was added Intermediate 4 (12.2 g, 30.3 mmol) and Cs2CO3 (28.68 g, 87.98 mmol). The solution was degassed with Ar for 10 min. Tetrakis(triphenylphosphine)palladium (5.00 g, 4.33 mmol) was added and the mixture was degassed for 10 min. The reaction was heated at 100 °C for 16 h. Upon completion, the reaction was passed through Celite, and the pad was washed with EtOAc (2 x 100 mL). The filtrate was washed with water (2 x 100 mL), brine (100 mL), dried (Na2SO4), filtered and concentrated. The residue was purified by flash chromatography (SiO2, 35- 50% EtOAc/Pet. ether) to afford Intermediate 20 (10 g, 34%) as an oil. MS (ESI) 582.4 [M+H]+. Intermediate 21 Methyl 3-(3-acetoxypropyl)-6-chloro-7-(1-ethyl-5-(hydroxymethyl)-3-methyl-1H-pyrazol-4-yl)- 1-methyl-1H-indole-2-carboxylate [0097] To a stirred solutio
Figure imgf000053_0002
0 (10.0 g, 17.6 mmol) in DCM (100 mL) was added TFA (13.14 mL, 19.59 mmol) at 0 oC. The ice bath was removed, and the reaction was stirred at rt for 2 h. Upon completion, the reaction was quenched with sat. NaHCO3 (100 mL) at 0 oC. The solid was collected by filtration and washed with water (100 mL). The solid was dissolved in DCM (500 mL), dried (Na2SO4) and filtered, and the solvent was evaporated. The residue was purified by flash chromatography (SiO2, 5% MeOH/DCM) to afford Intermediate 21 (6 g, 75%) as an off-white solid. MS (ESI) 462.3 [M+H]+. Intermediate 22 Methyl 3-(3-acetoxypropyl)-6-chloro-7-(5-(chloromethyl)-1-ethyl-3-methyl-1H-pyrazol-4-yl)-1- methyl-1H-indole-2-carboxylate [0098] To a stirred solutio
Figure imgf000054_0001
21 (3.00 g, 6.51 mmol) in DCM (30 mL) under argon was added SOCl2 (0.56 mL, 7.8 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 30 min. The solvent was evaporated, and the reaction was quenched with sat. NaHCO3 (50 mL). The mixture was extracted with DCM (2 x 50 mL). The combined organic layers were washed with brine (1 x 75 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 22 (3 g) as an oil. MS (ESI) 480.3 [M+H]+. Intermediate 23 Methyl 3-(3-acetoxypropyl)-6-chloro-7-(1-ethyl-5-(iodomethyl)-3-methyl-1H-pyrazol-4-yl)-1- methyl-1H-indole-2-carboxylate [0099] To a stirred solutio
Figure imgf000054_0002
22 (3.00 g, 6.25 mmol) in dry MeCN (30 mL) was added NaI (1.60 g, 10.7 mmol) at rt. The reaction was heated to 80 oC for 3 h. Upon completion, the solvent was evaporated, and the mixture was diluted with water (50 mL). The mixture was extracted with EtOAc (2 x 50 mL). The combined organic layers were dried (Na2SO4), filtered and evaporated to afford Intermediate 23 (3 g) as a semi solid. MS (LCMS) 572.3 [M+H]+. Intermediate 24 Methyl 6-chloro-7-(1-ethyl-5-((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H- pyrazol-3-yl)methyl)thio)methyl)-3-methyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1-methyl-1H- indole-2-carboxylate [0100] A suspen
Figure imgf000055_0001
mol) and K2CO3 (1.74 g, 12.608 mmol) in MeOH (30 mL) was degassed with Ar for 15 min. In a separate flask, Intermediate 15 (2.10 g, 5.25 mmol) in MeOH (30 mL) was degassed with Ar for 10 min. The solution of Intermediate 15 in MeOH was added to the previous reaction dropwise. The reaction was stirred at rt 1 h. The solvent was evaporated, and the reaction was diluted with water (30 mL) and extracted with EtOAc (3 x 30 mL). The combined the organic layers were washed with brine (90 mL), dried (Na2SO4), filtered and evaporated. The residue was purified by flash chromatography (SiO2, 50-70% EtOAc/Pet. ether) to afford Intermediate 24 (2.2 g, 59%) as an off-white solid. MS (ESI) 718.4 [M+H]+. Intermediate 25A (Ra)-(+)-Methyl (Z)-16-chloro-21-ethyl-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000055_0002
Intermediate 25B (Sa)-(-)-Methyl (Z)-16-chloro-21-ethyl-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0101] To a stirred, r
Figure imgf000056_0001
2.78 mmol) in DCM (10 mL) was added a solution of di-tert-butyl (E)-diazene-1,2-dicarboxylate (641 mg, 2.78 mmol) and Intermediate 24 (1.00 g, 1.39 mmol) in DCM (5 mL). The reaction was stirred at rt for 3 h. The reaction was quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated. Two equivalent reactions were run and combined for purification. The residue was purified by flash chromatography (SiO2, 90% EtOAc/Pet. ether) to afford racemic methyl (Z)-16-chloro-21-ethyl-11,23,61-trimethyl- 11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (1 g, 52%) as an off-white solid. MS (ESI) 700.5 [M+H]+. The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 40% CH3CN:MeOH; 4:1) to give peak 1 (Intermediate 25A, 200 mg) and peak 2 (Intermediate 25B, 200 mg). Intermediate 25A: off-white solid; 99.9% chiral purity; MS (LCMS) 700.5 [M+H]+. Intermediate 25B: off-white solid; 99.6% chiral purity; MS (LCMS) 700.5 [M+H]+. The absolute stereochemistry of Intermediate 25A and Intermediate 25B was arbitrarily assigned. Example 1A (Ra)-(+)-(Z)-16-Chloro-21-ethyl-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid
Figure imgf000056_0002
[0102] To a stirred solution of Intermediate 25A (200 mg, 0.285 mmol) in MeOH/THF/H2O (1:1:1, 6 mL) was added LiOH∙H2O (180 mg, 4.29 mmol) at rt. The reaction was stirred at 80 °C for 2 h. Upon completion, the solvent was evaporated. The aqueous layer was acidified to pH 6 using 2 N aq. HCl. The solid was filtered and washed with water (5 mL). The solid was dried under vacuum to afford Example 1A (189 mg, 96%) as an off-white solid. 99.7% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.30 (br s, 1H), 8.12 (d, J=7.6 Hz, 1H), 7.74-7.71 (m, 2H), 7.59-7.48 (m, 2H), 7.38 (s, 1H), 7.01 (d, J=8.4 Hz, 1H), 6.62 (s, 1H), 4.77 (s, 1H), 4.30-4.26 (m, 1H), 4.14-3.98 (m, 4H), 3.89-3.87 (m, 1H), 3.68 (s, 3H), 3.55-3.51 (m, 1H), 3.46 (s, 3H), 3.43- 3.41 (m, 1H), 3.24-3.06 (m, 3H), 2.95-2.91 (m, 1H), 2.33-2.32 (m, 1H), 2.22 (br s, 1H), 1.88 (s, 3H), 1.33 (t, J=7.2 Hz, 3H); MS (LCMS) 686.4 [M+H]+. The absolute stereochemistry of Example 1A was arbitrarily assigned. Example 1B (Sa)-(-)-(Z)-16-Chloro-21-ethyl-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0103] To a stirred
Figure imgf000057_0001
25B (200 mg, 0.285 mmol) in MeOH/THF/H2O (1:1:1, 6 mL) was added LiOH∙H2O (180 mg, 4.29 mmol) at rt. The reaction was stirred at 80 °C for 2 h. Upon completion, the solvent was evaporated. The aqueous layer was acidified to pH 6 using 2 N aq. HCl. The solid was filtered and washed with water (5 mL). The solid was dried under vacuum to afford Example 1B (167 mg, 85%) as an off-white solid.; 99.6% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.20 (br s, 1H), 8.12 (d, J=7.6 Hz, 1H), 7.75-7.70 (m, 2H), 7.51-7.44 (m, 2H), 7.36 (s, 1H), 7.01 (d, J=8.4 Hz, 1H), 6.62 (s, 1H), 4.77 (s, 1H), 4.35-4.25 (m, 1H), 4.20-3.98 (m, 4H), 3.89-3.87 (m, 1H), 3.69 (s, 3H), 3.58-3.41 (m, 5H), 3.22-3.12 (m, 3H), 2.95-2.92 (m, 1H), 2.36-2.34 (m, 1H), 2.23 (br s, 1H), 1.88 (s, 3H), 1.33 (t, J=7.2 Hz, 3H); MS (LCMS) 686.4 [M+H]+. The absolute stereochemistry of Example 1B was arbitrarily assigned. Intermediate 26 (1,3-Dimethyl-1H-pyrazol-5-yl)methanol [0104] To a stirred solution
Figure imgf000058_0001
imethyl-1H-pyrazole-5-carboxylate (25.0 g, 149 mmol) in THF (200 mL) was added 2.4 M LAH in THF (74.0 mL, 179 mmol) at 0°C. The reaction was stirred under N2 for 3 h at 0 oC. The reaction was quenched with sat. Na2SO4 (50 mL) at 0 oC. The white solid was filtered and washed with 10% MeOH in DCM (500 mL). The filtrate was dried (Na2SO4) and evaporated to afford Intermediate 26 (16 g, 85%) as a colorless oil. MS (LCMS) 127.1 [M+H]+. Intermediate 27 (4-Bromo-1,3-dimethyl-1H-pyrazol-5-yl)methanol [0105] NBS (27.1 g, 152 mm d portion wise with stirring t o
Figure imgf000058_0002
o a 0 C solution of Intermediate 26 (16.0 g, 127 mmol) in DMF (80 mL). The reaction was stirred under N2 for 2 h at rt. Upon completion by LCMS, the reaction was quenched with cold water (500 mL). The white solid was collected by filtration and washed with water (200 mL). The solid was dried under high vacuum to afford Intermediate 27 (17.0 g, 66%) as a yellow solid. MS (LCMS) 204.9 [M+H]+. Intermediate 28 4-Bromo-5-(((4-methoxybenzyl)oxy)methyl)-1,3-dimethyl-1H-pyrazole [0106] To a stirred, 0 oC sol
Figure imgf000058_0003
ediate 27 (12.0 g, 58.5 mmol) in DMF (60 ml) was added KI (4.80 g, 29.2 mmol) and NaH (5.60 g, 117 mmol). The reaction was stirred at 0°C for 20 min. PMBCl (11.8 mL, 87.8 mmol) was added dropwise over 5 min. The reaction was then stirred at rt for 2 h. Upon completion by LCMS, the reaction was quenched with cold water (200 mL). The resulting solid was collected by filtration and washed with water (200 mL). The crude material was purified by flash chromatography (SiO2, 10% EtOAc/Pet. ether) to afford Intermediate 28 (12 g, 63%) as an off-white solid. MS (LCMS) 325.1 [M+H]+. Intermediate 29
5-(((4-Methoxybenzyl)oxy)methyl)-l,3-dimethyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-lH-pyrazole
Figure imgf000059_0001
[0107] 2.5 M n-BuLi in hexanes (14.8 mL, 37.0 mmol) was added dropwise to a -78
°C solution of Intermediate 28 (10.0 g, 30.8 mmol) in THF (100 mL). The reaction was stirred at -78 °C for 1 h. 2-Isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (8.0 g, 43 mmol) was added dropwise, and the reaction was stirred for 3 h at -78 °C. Upon completion, the reaction was quenched with EtOAc (20 mL) and concentrated. The crude mixture was suspended in EtOAc (200 mL) and filtered through a Celite pad which was washed with EtOAc (100 mL). The solvent was removed, and the residue was purified by flash chromatography (SiO2, 10% EtOAc/Pet. ether) to afford Intermediate 29 (6.8 g, 60%) as an oil. MS (LCMS) 373.3 [M+H]+.
Intermediate 30
Methyl 7-bromo-6-fluoro-3-(3-methoxy-3-oxopropyl)-lH-indole-2-carboxylate
Figure imgf000059_0002
[0108] Intermediate 30 was synthesized from 2-bromo-3-fluoroaniline following a similar procedure for the preparation of Intermediate 1. MS (LCMS) 358.1 [M+H]+.
Intermediate 31
Methyl 3-(3-acetoxypropyl)-7-bromo-6-fluoro-l-methyl-lH-indole-2-carboxylate
Figure imgf000059_0003
[0109] Intermediate 31 was synthesized from Intermediate 30 following similar procedures for the preparation of Intermediates 2-4. MS (LCMS) 386.2 [M+H]+. Intermediate 32 Methyl 3-(3-acetoxypropyl)-6-fluoro-7-(5-(((4-methoxybenzyl)oxy)methyl)-1,3-dimethyl-1H- pyrazol-4-yl)-1-methyl-1H-indole-2-carboxylate [0110] Intermediate 32 w
Figure imgf000060_0001
m Intermediate 31 and Intermediate 29 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 552.4 [M+H]+. Intermediate 33 Methyl 3-(3-acetoxypropyl)-6-fluoro-7-(5-(hydroxymethyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1- methyl-1H-indole-2-carboxylate [0111] Intermediate 33
Figure imgf000060_0002
om Intermediate 32 following a similar procedure for the preparation of Intermediate 21. MS (LCMS) 432.4 [M+H]+. Intermediate 34 Methyl 3-(3-acetoxypropyl)-7-(5-(chloromethyl)-1,3-dimethyl-1H-pyrazol-4-yl)-6-fluoro-1- methyl-1H-indole-2-carboxylate [0112] Intermediate 34
Figure imgf000060_0003
om Intermediate 33 following a similar procedure for the preparation of Intermediate 22. MS (LCMS) 450.3 [M+H]+. Intermediate 35 Methyl 3-(3-acetoxypropyl)-6-fluoro-7-(5-(iodomethyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1- methyl-1H-indole-2-carboxylate [0113] Intermediate 35
Figure imgf000061_0001
om Intermediate 34 following a similar procedure for the preparation of Intermediate 23. MS (LCMS) 542.3 [M+H]+. Intermediate 36 Methyl 6-fluoro-7-(5-((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H-pyrazol-3- yl)methyl)thio)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1-methyl-1H- indole-2-carboxylate [0114] Intermed
Figure imgf000061_0002
mediate 35 and Intermediate 15 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 688.9 [M+H]+. Intermediate 37A (Ra)-(+)-Methyl (Z)-16-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000061_0003
Intermediate 37B (Sa)-(-)-Methyl (Z)-16-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0115] Intermediates
Figure imgf000062_0001
ized from Intermediate 36 following a similar procedure for the preparation of Intermediates 25A and 25B to give racemic methyl (Z)- 16-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola-2(4,5),6(3,5)- dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (270 mg, 28%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 40% CH3CN:MeOH; 4:1) to give peak 1 (Intermediate 37A, 110 mg) and peak 2 (Intermediate 37B, 110 mg). Intermediate 37A: off-white solid; 99.9% chiral purity; MS (LCMS) 670.2 [M+H]+. Intermediate 37B: off-white solid; 99.3% chiral purity; MS (LCMS) 670.5 [M+H]+. The absolute stereochemistry of Intermediate 37A and Intermediate 37B was arbitrarily assigned. Example 2A (Ra)-(+)-(Z)-16-Fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0116] Example 2A
Figure imgf000062_0002
ermediate 37A following a similar procedure for the preparation of Example 1. Example 2A: (52 mg, 50%), off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.25 (br s, 1H), 8.13 (d, J=7.6 Hz, 1H), 7.80- 7.70 (m, 2H), 7.54-7.45 (m, 2H), 7.37 (s, 1H), 6.78 (t, J=9.0 Hz, 1H), 6.66 (s, 1H), 4.70 (s, 1H), 4.19 (q, J=16.2 Hz, 2H), 4.10-1.00 (m, 1H), 3.90-3.87 (m, 1H), 3.76 (s, 3H), 3.70 (s, 3H), 3.55- 3.40 (m, 5H), 3.33-3.25 (m, 1H), 3.20-3.10 (m, 2H), 2.96 (d, J=14.4 Hz, 1H), 2.45-2.37 (m, 1H), 2.30-2.20 (m, 1H), 1.90 (s, 3H); MS (LCMS) 656.3 [M+H]+. The absolute stereochemistry of Example 2A was arbitrarily assigned. Example 2B (Sa)-(-)-(Z)-16-Fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0117] Example 2B
Figure imgf000063_0001
ermediate 37B following a similar procedure for the preparation of Example 1. Example 2B: (55 mg, 55%), off-white solid; 99.3% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.25 (br s, 1H), 8.15-8.10 (m, 1H), 7.81-7.71 (m, 2H), 7.54-7.45 (m, 2H), 7.37 (s, 1H), 6.78 (t, J=9.2 Hz, 1H), 6.66 (s, 1H), 4.70 (s, 1H), 4.19 (q, J=16.2 Hz, 2H), 4.10-1.00 (m, 1H), 3.90-3.87 (m, 1H), 3.76 (s, 3H), 3.70 (s, 3H), 3.52 (s, 3H), 3.50-3.40 (m, 2H), 3.33-3.25 (m, 1H), 3.15-3.10 (m, 2H), 2.96 (d, J=14.4 Hz, 1H), 2.45-2.37 (m, 1H), 2.30-2.20 (m, 1H), 1.90 (s, 3H); MS (LCMS) 656.3 [M+H]+. The absolute stereochemistry of Example 2B was arbitrarily assigned. Intermediate 38 Methyl 7-bromo-3-(3-methoxy-3-oxopropyl)-6-methyl-1H-indole-2-carboxylate [0118] Intermediate 38
Figure imgf000063_0002
m 2-bromo-3-methylaniline following a similar procedure for the preparation of Intermediate 1. MS (LCMS) 354.2 [M+H]+. Intermediate 39 Methyl 3-(3-acetoxypropyl)-7-bromo-1,6-dimethyl-1H-indole-2-carboxylate
Figure imgf000063_0003
[0119] Intermediate 39 was synthesized from Intermediate 38 following similar procedures for the preparation of Intermediates 2-4. MS (LCMS) 382.4 [M+H]+.
Intermediate 40
Methyl 3-(3-acetoxypropyl)-7-(5-(((4-methoxybenzyl)oxy)methyl)-l,3-dimethyl-lH-pyrazol-4- yl)- 1 ,6-dimethyl- lH-indole-2-carboxylate
Figure imgf000064_0001
[0120] Intermediate 40 was synthesized from Intermediate 39 and Intermediate 29 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 548.9.
Intermediate 41
Methyl 3-(3-acetoxypropyl)-7-(5-(hydroxymethyl)-l,3-dimethyl-lH-pyrazol-4-yl)-l,6-dimethyl- lH-indole-2-carboxylate
Figure imgf000064_0002
[0121] Intermediate 41 was synthesized from Intermediate 40 following a similar procedure for the preparation of Intermediate 21. MS (LCMS) 428.4 [M+H]+.
Intermediate 42
Methyl 3-(3-acetoxypropyl)-7-(5-(chloromethyl)-l,3-dimethyl-lH-pyrazol-4-yl)-l,6-dimethyl- lH-indole-2-carboxylate
Figure imgf000064_0003
[0122] Intermediate 42 was synthesized from Intermediate 41 following a similar procedure for the preparation of Intermediate 22. MS (LCMS) 446.8 [M+H]+. Intermediate 43 Methyl 3-(3-acetoxypropyl)-7-(5-(iodomethyl)-1,3-dimethyl-1H-pyrazol-4-yl)-1,6-dimethyl-1H- indole-2-carboxylate [0123] Intermediate 43 w
Figure imgf000065_0001
om Intermediate 42 following a similar procedure for the preparation of Intermediate 23. MS (LCMS) 538.3 [M+H]+. Intermediate 44 Methyl 7-(5-((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H-pyrazol-3- yl)methyl)thio)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1,6-dimethyl-1H- indole-2-carboxylate [0124] Intermed
Figure imgf000065_0002
mediate 43 and Intermediate 15 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 684.4 [M+H]+. Intermediate 45A (Ra)-(+)-Methyl (Z)-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000065_0003
Intermediate 45B (Sa)-(-)-Methyl (Z)-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0125] Intermediates
Figure imgf000066_0001
ized from Intermediate 44 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola-2(4,5),6(3,5)- dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (600 mg, 15%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 40% CH3CN:MeOH; 4:1) to give peak 1 (Intermediate 45A, 210 mg) and peak 2 (Intermediate 45B, 200 mg). Intermediate 45A: off-white solid; 99.9% chiral purity; MS (LCMS) 666.5 [M+H]+. Intermediate 45B: off-white solid; 99.8% chiral purity; MS (LCMS) 666.5 [M+H]+. The absolute stereochemistry of Intermediate 45A and Intermediate 46B was arbitrarily assigned. Example 3A (Ra)-(+)-(Z)-11,16,21,23,61-Pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0126] Example 3A
Figure imgf000066_0002
ermediate 45A following a similar procedure for the preparation of Example 1. Example 3A: (175 mg, 90%), off-white solid; 99.8% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.15 (brs, 1H), 8.17 (d, J=7.2 Hz, 1H), 7.75 (d, J=7.2 Hz, 1H), 7.60-7.44 (m, 4H), 6.84-6.81 (m, 1H), 6.52 (s, 1H), 4.71 (s, 1H), 4.25 (d, J=15.6 Hz, 1H), 4.11-4.07 (m, 1H), 3.91-3.87 (m, 2H), 3.74 (s, 3H), 3.68 (s, 3H), 3.62-3.58 (m, 1H), 3.50- 3.40 (m, 4H), 3.17-3.13 (m, 2H), 3.06-3.02 (m, 1H), 2.84 (d, J=14.0 Hz, 1H), 2.33 (m, 1H), 2.18 (m, 1H), 1.94 (s, 3H), 1.86 (s, 3H); MS (LCMS) 652.4 [M+H]+. The absolute stereochemistry of Example 3A was arbitrarily assigned. Example 3B (Sa)-(-)-(Z)-11,16,21,23,61-Pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0127] Example 3B
Figure imgf000067_0001
ermediate 45B following a similar procedure for the preparation of Example 1. Example 3B: (177 mg, 90%), off-white solid; 99.8% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.12 (s, 1H), 8.22-8.15 (m, 1H), 7.80-7.73 (m, 1H), 7.59 (d, J=8.4 Hz, 1H), 7.52-7.49 (m, 2H), 7.44 (s, 1H), 6.84 (d, J=8.4 Hz, 1H), 6.52 (s, 1H), 4.71 (s, 1H), 4.25 (d, J=8.4 Hz, 1H), 4.09 (d, J=15.6 Hz, 1H), 3.91-3.85 (m, 2H), 3.74 (s, 3H), 3.68 (s, 3H), 3.51-3.47 (m, 4H), 3.25-3.13 (m, 2H), 3.04 (d, J=14.0 Hz, 1H), 2.84 (d, J=14.0 Hz, 1H), 2.34 (bs, 1H), 2.01 (bs, 1H), 1.94 (s, 3H) 1.83 (s, 3H); MS (LCMS) 652.4 [M+H]+. The absolute stereochemistry of Example 3B was arbitrarily assigned. Intermediate 46 Ethyl 5-(hydroxymethyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate [0128] To a stirred solution
Figure imgf000067_0002
te 5 (30.0 g, 73.5 mmol) in THF (300 mL) was added NaHMDS (44.0 mL, 88.2 mmol) and NaI (5.50 g, 36.6 mmol) at 0 °C. The reaction was stirred for 30 min. PMBCl (13.85 g, 88.23 mmol) was added at 0 °C. The reaction was stirred at 50 °C for 2 h. Upon completion by TLC, the reaction was quenched with sat. NH4Cl (100 mL) and extracted with EtOAc (2 x 150 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated. The crude residue was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford ethyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-1-(4-methoxybenzyl)- 1H-pyrazole-3-carboxylate (34.5 g, 89%) as an oil. (LCMS) 529.5 [M+H] +. [0129] To a stirred solution of ethyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-1-(4- methoxybenzyl)-1H-pyrazole-3-carboxylate (34.5 g, 65.3 mmol) in THF (345 mL) was added CsF (19.86 g, 130.7 mmol) and 1M TBAF in THF (78.4 mL, 78.4 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 4 h. Upon completion by TLC, the reaction was quenched with NH4Cl (250 mL) at 0 °C and extracted with EtOAc (2 x 500 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated. The crude residue was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford Intermediate 46 (17 g, 90%) as an oil. (LCMS) 291.3 [M+H] +. Intermediate 47 Ethyl 5-(chloromethyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate [0 under Ar
Figure imgf000068_0001
The volatiles were evaporated and sat. NaHCO3 (150 mL) was added. The mixture was extracted with DCM (2 x 150 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated to afford Intermediate 47 (10 g, 85%) as an oil. MS (LCMS) 309.5 [M+H]+. Intermediate 48 Methyl 5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1-(4-methoxybenzyl)-1H-pyrazole-3- carboxylate OH
Figure imgf000068_0002
g, 35.7 mmol) was dissolved in MeOH (50 mL) and degassed with Ar for 10 min. The solution of Intermediate 11 was added dropwise to the mixture of Intermediate 47 and K2CO3, and the reaction was stirred at rt for 16 h. The reaction was concentrated and diluted with water (100 mL). The mixture was extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (200 mL), dried (Na2SO4), filtered and concentrated. The crude mixture was triturated with a mixture of DCM (25 mL) and n-pentane (25 mL) to afford Intermediate 48 (7 g, 49%). MS (LCMS) 435.3 [M+H]+. Intermediate 49 3-(((3-(Hydroxymethyl)-1-(4-methoxybenzyl)-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-ol [0132] To a stirre mmol) in THF (140 mL)
Figure imgf000069_0003
was added 25% DIBAL in PhMe (11.45 g, 80.64 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction cooled to 0 oC and quenched with sat. NH4Cl (50 mL). The slurry was filtered through Celite which was washed with EtOAc (200 mL). The filtrate was washed with brine (200 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 49 (11 g, 84%) as an off-white solid. MS (LCMS) 405.3 [M+H]+. Intermediate 50 3-(((3-(Chloromethyl)-1-(4-methoxybenzyl)-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-ol OH [0133] To a stirre mmol) in DCM (90 mL)
Figure imgf000069_0002
under Ar was added SOCl2 (2.47 mL, 33.9 mmol) at 0 °C, and the reaction was stirred at rt for 3 h. The reaction was diluted with DCM (150 mL) and washed with sat. NaHCO3 (3 x 100 mL). The organic layer was separated, dried (Na2SO4), filtered and evaporated to afford Intermediate 50 (9 g, 96%) as an oil. MS (LCMS) 425.3[M+H]+. Intermediate 51 3-(((3-(Chloromethyl)-1-(4-methoxybenzyl)-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-yl acetate OAc [0134] To a g, 23.6 mmol) in CH3CN (50
Figure imgf000069_0001
mL), was added TEA (6.58 mL, 47.2 mmol), Ac2O (2.89 mL, 30.7 mmol) and DMAP (280 mg, 2.36 mmol) at 0 °C. The reaction was stirred at rt for 2 h. Upon completion by TLC, the solvent was evaporated and water (200 mL) was added. The mixture was extracted with EtOAc (2 x 200 mL). The combined organic layers were separated, dried (Na2SO4), and the solvent evaporated. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 51 (10 g, 90%) as an oil. MS (LCMS) 467.2 [M+H]+. Intermediate 52 3-(((3-((Acetylthio)methyl)-1-(4-methoxybenzyl)-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-yl acetate [0135] To a stirred sol
Figure imgf000070_0001
(10.0 g, 21.5 mmol) in CH3CN (100 mL) was added KSAc (3.92 g, 34.3 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the solvent was evaporated and water (200 mL) was added. The mixture was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (400 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 52 (10 g, 90%) as an oil. MS (LCMS) 507.5 [M+H]+. Intermediate 53 Methyl 6-chloro-3-(3-methoxy-3-oxopropyl)-7-(5-(((4-methoxybenzyl)oxy)methyl)-1,3- dimethyl-1H-pyrazol-4-yl)-1H-indole-2-carboxylate [0136] Intermediate 53
Figure imgf000070_0002
m Intermediate 1 and Intermediate 29 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 540.7 [M+H]+. Intermediate 54 Methyl 6-chloro-3-(3-methoxy-3-oxopropyl)-7-(5-(((4-methoxybenzyl)oxy)methyl)-1,3- dimethyl-1H-pyrazol-4-yl)-1-methyl-1H-indole-2-carboxylate O OMe [01 F (90 mL) was a
Figure imgf000071_0003
dded Cs2CO3 (7.20 g, 22.2 mmol) and MeI (1.80 mL, 29.6 mmol) at 0 C. The ice bath was removed, and the reaction was stirred at rt for 1 h. The reaction was quenched with water (150 mL) and extracted with EtOAc (2 x 150 mL). The combined organic layers were washed with brine (300 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 54 (8 g) as an oil. MS (LCMS) 554.4 [M+H]+. Intermediate 55 Methyl 6-chloro-7-(5-(hydroxymethyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-methoxy-3- oxopropyl)-1-methyl-1H-indole-2-carboxylate [0138] Intermediate 55
Figure imgf000071_0001
Intermediate 54 following a similar procedure for the preparation of Intermediate 21. MS (LCMS) 434.4 [M+H]+. Intermediate 56 Methyl 6-chloro-7-(5-(chloromethyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-methoxy-3- oxopropyl)-1-methyl-1H-indole-2-carboxylate
Figure imgf000071_0002
[0139] Intermediate 56 was prepared from Intermediate 55 following a similar procedure for the preparation of Intermediate 22. MS (LCMS) 452.4 [M+H]+.
Intermediate 57
Methyl 6-chloro-7-(5-(iodomethyl)-l,3-dimethyl-lH-pyrazol-4-yl)-3-(3-methoxy-3-oxopropyl)-
1 -methyl- lH-indole-2-carboxylate
Figure imgf000072_0001
[0140] Intermediate 57 was prepared from Intermediate 56 following a similar procedure for the preparation of Intermediate 23. MS (LCMS) 544.7 [M+H]+.
Intermediate 58
Methyl 6-chloro-7-(5-((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-l-(4-methoxybenzyl)-lH- pyrazol-3-yl)methyl)thio)methyl)-l,3-dimethyl-lH-pyrazol-4-yl)-3-(3-methoxy-3-oxopropyl)-l- methyl- lH-indole-2-carboxylate
Figure imgf000072_0002
[0141] Intermediate 58 was prepared from Intermediate 57 and Intermediate 52 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 838.6 [M+H]+.
Intermediate 59
Methyl 6-chloro-7-(5-((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-l-(4-methoxybenzyl)-lH- pyrazol-3-yl)methyl)thio)methyl)-l,3-dimethyl-lH-pyrazol-4-yl)-3-(3-hydroxypropyl)-l-methyl- lH-indole-2-carboxylate
Figure imgf000072_0003
[0142] To a suspension of Intermediate 58 (1.20 g, 1.43 mmol) in dry THF (20 mL) was added 1.0 M BH3∙THF in THF (7.80 mL, 7.80 mmol) dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was quenched with MeOH (15 mL) and 6N HCl (15 mL) at 0 oC. The mixture was stirred at rt for 30 min. The reaction was diluted with water (75 mL) and extracted with 10% MeOH in DCM (2 x 150 ml). The organic layer was washed with sat. NaHCO3 (2 x 150 mL), dried (Na2SO4), filtered and evaporated. The reaction was run again on an equivalent scale and combined for purification. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 59 (1.2 g, 50%) as a white solid. MS (LCMS) 810.6 [M+H]+. Intermediate 60 Methyl (Z)-16-chloro-61-(4-methoxybenzyl)-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0143] Intermediate
Figure imgf000073_0001
termediate 59 following a similar procedure for the preparation of Intermediates 25A and 25B. MS (LCMS) 792.6 [M+H]+. Intermediate 61A (Ra)-(+)-Methyl (Z)-16-chloro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000073_0002
Intermediate 61B (Sa)-(-)-Methyl (Z)-16-chloro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0144] Intermediate
Figure imgf000074_0001
was added to a sealed tube followed by TFA (10 mL) and anisole (817 mg, 7.57 mmol). The tube was sealed and then stirred at 120 oC for 20 h. Upon completion by LCMS, the reaction was concentrated and diluted with EtOAc (30 mL). The organic layer was washed with sat. NaHCO3 (30 mL), brine (30 mL) and dried (Na2SO4). The mixture was filtered and concentrated. The crude material was purified by flash chromatography (SiO2, 40% EtOAc/Pet. ether) to afford racemic methyl (Z)-16-chloro-11,21,23- trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)- naphthalenacyclo-tridecaphane-12-carboxylate (280 mg, 55%) as an off-white solid. The atropisomers were separated by chiral SFC chromatography (Chiralcel-AS-3 (30 x 250 mm), 30% (0.2% NH3 in MeOH)) to give peak 1 (Intermediate 61A, 110 mg) and peak 2 (Intermediate 61B, 90 mg). Intermediate 61A: off-white solid; 98.9% chiral purity; MS (LCMS) 672.3 [M+H]+. Intermediate 61B: off-white solid; 98.9% chiral purity; MS (LCMS) 672.3 [M+H]+. The absolute stereochemistry of Intermediate 61A and Intermediate 61B was arbitrarily assigned. Example 4A (Ra)-(+)-(Z)-16-Chloro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0145] Example 4A
Figure imgf000074_0002
ermediate 61A following a similar procedure for the preparation of Example 1. Example 4A: (45 mg, 42%), off-white solid; 99.3% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 12.9 (br s, 2H), 8.15 (d, J=8.0 Hz, 1H), 7.74 (d, J=7.2 Hz, 1H), 7.70-7.65 (m, 1H), 7.53-7.45 (m, 2H), 7.37 (s, 1H), 6.98-6.92 (m, 1H), 6.66 (s, 1H), 4.69 (s, 1H), 4.15-3.95 (m, 3H), 3.90-3.82 (m, 1H), 3.78 (s, 3H), 3.60-3.40 (m, 5H), 3.33- 3.25 (m, 2H) 3.20-3.10 (m, 1H), 3.05-2.95 (m, 1H), 2.40-2.30 (m, 1H), 2.25-2.15 (m, 1H), 1.87 (s, 3H); MS (LCMS) 658.3 [M+H]+. The absolute stereochemistry of Example 4A was arbitrarily assigned. Example 4B (Sa)-(-)-(Z)-16-Chloro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0146] Example 4B
Figure imgf000075_0001
ermediate 61B following a similar procedure for the preparation of Example 1. Example 4B: (50 mg, 56%), white solid; 98.6% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 12.9 (br s, 2H), 8.15 (d, J=8.0 Hz, 1H), 7.74 (d, J=7.2 Hz, 1H), 7.70-7.65 (m, 1H), 7.53-7.45 (m, 2H), 7.37 (s, 1H), 6.98-6.92 (m, 1H), 6.66 (s, 1H), 4.69 (s, 1H), 4.15-3.95 (m, 3H), 3.90-3.82 (m, 1H), 3.78 (s, 3H), 3.60-3.40 (m, 5H), 3.33- 3.25 (m, 2H), 3.20-3.10 (m, 1H), 3.05-2.95 (m, 1H), 2.40-2.30 (m, 1H), 2.25-2.15 (m, 1H), 1.87 (s, 3H); MS (LCMS) 658.3 [M+H]+. The absolute stereochemistry of Example 4B was arbitrarily assigned. Intermediate 62 Ethyl 4-oxo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate [0147] To a stirred solution
Figure imgf000075_0002
H-pyrazole-3,5-dicarboxylate (20.0 g, 94.3 mmol) in DMF (200 mL) was added Cs2CO3 (36.9 g, 113 mmol) and tert-butyl (2-bromoethyl) carbamate (26.29 g, 117.9 mmol) at 0 oC. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was diluted with water (2 L). The precipitated solid was collected by filtration, washed with water (2 x 1 L) and dried under high vacuum. The solid was triturated with pentane and dried under high vacuum to afford diethyl 1-(2-((tert- butoxycarbonyl)amino)ethyl)-1H-pyrazole-3,5-dicarboxylate (20 g, 60%) as a white solid. MS (LCMS) 356.4 [M+H]+. [0148] Diethyl 1-(2-((tert-butoxycarbonyl)amino)ethyl)-1H-pyrazole-3,5- dicarboxylate (20.0 g, 56.3 mmol) was added to 4N HCl in dioxane (200 mL) at 0 oC and stirred at rt for 4 h. Upon completion by TLC, the solvent was evaporated. The mixture was diluted with sat. Na2CO3 (200 mL), stirred at rt for 1 h, and extracted with DCM (3 x 500 mL). The combined organic layers were dried (Na2SO4), filtered and evaporated. The crude mixture was triturated with pentane and dried to afford Intermediate 62 (10 g, 84%) as a white solid. MS (LCMS) 210.2 [M+H]+. Intermediate 63 tert-Butyl 2-(hydroxymethyl)-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate [0149] To a stirred solu
Figure imgf000076_0001
2 (28.0 g, 134 mmol) in THF (280 ml) was added 2.4 M LiAlH4 in THF (223 mL, 535 mmol) at 0 ^C. The ice bath was removed, and the reaction was stirred at 70 oC for 6 h. Upon completion by TLC, the reaction was cooled to 0°C, diluted with EtOAc (500 mL) and diluted by the dropwise addition of water (29.2 mL). 15% NaOH (29.2 mL) and water (87.6 mL) was added, and the mixture was stirred at rt for 15 min. MgSO4 was added, and the mixture was stirred for 15 min. The mixture was filtered through Celite, and the pad was washed with EtOAc (2 x 100 mL). The filtrate was washed with brine (500 mL), dried (Na2SO4), filtered and evaporated to afford (4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazin-2-yl)methanol (16 g, 78%) as an oil. MS (LCMS) 154.2 [M+H]+. [0150] To a stirred solution of (4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2- yl)methanol (18.0 g, 117 mmol) and triethylamine (27.7 g, 234 mmol) in DCM (180 mL) was added di-tert-butyl dicarbonate (30.8 g, 141 mmol) at 0 °C. The reaction was stirred at rt for 1 h. The reaction was diluted with water (300 mL) and extracted with DCM (3 x 200 mL). The combined organic layers were washed with brine (500 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 63 (18 g, 62%) as a white solid. MS (LCMS) 254.4 [M+H]+. Intermediate 64 tert-Butyl 3-bromo-2-(((4-methoxybenzyl)oxy)methyl)-6,7-dihydropyrazolo[1,5-a]pyrazine- 5(4H)-carboxylate [0151] To a stirred solu
Figure imgf000077_0001
(24.0 g, 94.8 mmol) in DCM (240 mL) was added NBS (20.2 g, 114 mmol) at 0 °C and the reaction was stirred at rt for 1 h. Upon completion by TLC, the reaction was diluted with sat. NaHCO3 (250 mL) and extracted with DCM (2 x 200 mL). The organic layer was separated, dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford tert- butyl 3-bromo-2-(hydroxymethyl)-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate (28 g, 76%) as an off-white solid. MS (LCMS) 332.2 [M+H]+. [0152] To a stirred solution of tert-butyl 3-bromo-2-(hydroxymethyl)-6,7- dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate (45.0 g, 136 mmol) in DMF (450 mL) was added 60 % NaH (6.86 g, 204 mmol) at 0 °C. The reaction was stirred at rt for 20 min. 1- (Chloromethyl)-4-methoxybenzene (31.9 g, 204 mmol) and KI (2.26 g, 13.6 mmol) was added, and the reaction was stirred at rt for 6 h. Upon completion by TLC, the reaction was quenched with sat. NH4Cl (250 ml). The mixture was extracted with EtOAc (4 x 250 mL). The combined organic layers were washed with water (2 x 250 mL), brine (250 mL), dried (Na2SO4) and concentrated. The crude material was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford Intermediate 64 (15 g, 75%) as a white solid. MS (LCMS) 452.3 [M+H]+. Intermediate 65 tert-Butyl 2-(((4-methoxybenzyl)oxy)methyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate [0153] To a stirred sol
Figure imgf000077_0002
(5.00 g, 11.1 mmol) in THF (50 mL) was added 1.6 M n-BuLi in hexanes (8.30 mL, 13.3 mmol) at -78 °C, and the reaction was stirred at -78 °C for 50 min. 2-Isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.09 g, 16.6 mmol) was added at -78 °C, and the reaction was stirred at -78 °C for 1 h. The solvents were evaporated, and the mixture was diluted with EtOAc (200 mL). The mixture was filtered through Celite, and the pad was washed with EtOAc (100 mL). The filtrate was evaporated, and the crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 65 (2.4 g, 43%) as a white solid. MS (LCMS) 500.5 [M+H]+. Intermediate 66 tert-Butyl 3-(3-(3-acetoxypropyl)-6-chloro-2-(methoxycarbonyl)-1-methyl-1H-indol-7-yl)-2- (((4-methoxybenzyl)oxy)methyl)-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate [0154] Intermediate 6
Figure imgf000078_0001
Intermediate 4 and Intermediate 65 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 695.6 [M+H]+. Intermediate 67 tert-Butyl 3-(3-(3-acetoxypropyl)-6-chloro-2-(methoxycarbonyl)-1-methyl-1H-indol-7-yl)-2- (hydroxymethyl)-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate [0155] To a stirred solut
Figure imgf000078_0002
(2.00 g, 2.87 mmol) in DCM (38 mL) and water (2 mL) was added DDQ (0.78 g, 3.4 mmol) at 0°C. The reaction was stirred at rt for 3 h. The reaction was quenched with sat. NaHCO3 (20 mL) and extracted with DCM (3 x 30 mL). The combined organic layers were washed with water (3 x 20 mL), brine (20 mL), dried (Na2SO4), filtered and concentrated. The reaction was run twice on an equivalent scale, and the reactions were combined for purification. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 67 (2.2 g, 66%) as an oil. MS (LCMS) 575.9 [M+H]+. Intermediate 68 tert-Butyl 3-(3-(3-acetoxypropyl)-6-chloro-2-(methoxycarbonyl)-l-methyl-lH-indol-7-yl)-2- (chloromethy 1) -6 ,7 -dihy dropyrazolo [ 1 , 5 - a] pyrazine-5 (4H) -c arboxy late
Figure imgf000079_0001
[0156] Intermediate 68 was synthesized from Intermediate 67 following a similar procedure from the preparation of Intermediate 22. MS (LCMS) 593.2 [M+H]+.
Intermediate 69 tert-Butyl 3-(3-(3-acetoxypropyl)-6-chloro-2-(methoxycarbonyl)-l-methyl-lH-indol-7-yl)-2- (iodomethyl)-6,7-dihydropyrazolo[l,5-a]pyrazine-5(4H)-carboxylate
Figure imgf000079_0002
[0157] Intermediate 69 was synthesized from Intermediate 68 following a similar procedure from the preparation of Intermediate 23. MS (LCMS) 685.4 [M+H]+.
Intermediate 70 tert-Butyl 3-(6-chloro-3-(3-hydroxypropyl)-2-(methoxycarbonyl)-l-methyl-lH-indol-7-yl)-2- ((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-l-methyl-lH-pyrazol-3-yl)methyl)thio)methyl)- 6,7-dihydropyrazolo[l,5-a]pyrazine-5(4H)-carboxylate
Figure imgf000079_0003
[0158] Intermediate 70 was synthesized from Intermediate 69 and Intermediate 15 following a similar procedure from the preparation of Intermediate 24. MS (LCMS) 831.5 [M+H]+. Intermediate 71 25-(tert-butyl) 12-methyl (Z)-16-chloro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8- dithia-2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12,25-dicarboxylate [0159] Intermedia
Figure imgf000080_0001
ermediate 70 following a similar procedure from the preparation of Intermediates 25A and 25B. MS (LCMS) 813.9 [M+H]+. Intermediate 72 Methyl (Z)-16-chloro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8-dithia-2(3,2)- pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)-naphthalenacyclo tridecaphane-12- carboxylate hydrochloride [0160] To a stirred
Figure imgf000080_0002
0 mg, 1.04 mmol) was added 4N HCl in dioxane (10 mL) at 0 °C. The reaction was stirred at rt for 2 h. The solvent was evaporated, and the crude material was triturated with a mixture of pentane (5 mL) and ether (5 mL) to afford Intermediate 72 (900 mg) as an off-white solid. MS (LCMS) 713.3 [M+H]+. Intermediate 73A (Ra)-(+)-Methyl (Z)-16-chloro-25-ethyl-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa- 4,8-dithia-2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000080_0003
Intermediate 73B (Sa)-(-)-Methyl (Z)-16-chloro-25-ethyl-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8- dithia-2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate [0161] To a stirred
Figure imgf000081_0001
mg, 1.26 mmol) in MeOH (8 mL) was added THF (2 mL), TEA (0.87 mL, 6.3 mmol), acetaldehyde (445 mg, 10.2 mmol) and NaCNBH3 (396 mg, 6.30 mmol) at rt. The reaction was stirred at rt for 16 h. The solvent was evaporated and water (20 mL) was added. The mixture was extracted with 3% MeOH/DCM (2 x 20 mL). The combined organic layers were dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 0.1% MeOH/DCM) to afford racemic methyl (Z)-16-chloro-25-ethyl-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (690 mg, 73%) as a white solid. MS (LCMS) 741.5 [M+H]+. The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 30% (0.5% DEA in MeOH)) to give peak 1 (Intermediate 73A, 280 mg) and peak 2 (Intermediate 73B, 280 mg). Intermediate 73A: off-white solid; 98.1% chiral purity; MS (LCMS) 741.6 [M+H]+. Intermediate 73B: off-white solid; 99.3% chiral purity; MS (LCMS) 741.6 [M+H]+. The absolute stereochemistry of Intermediate 73A and Intermediate 73B was arbitrarily assigned.
Example 5A (Ra)-(+)-(Z)-16-Chloro-25-ethyl-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0162] Example 5
Figure imgf000082_0001
mediate 73A following a similar procedure for the preparation of Example 1. Example 5A: (190 mg, 70%), off-white solid; 98.6% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.39 (br s, 1H), 11.8-11.0 (m, 1H), 8.17-8.08 (m, 1H), 7.95-7.80 (m, 1H), 7.78-7.70 (m, 1H), 7.54-7.43 (m, 2H), 7.38 (s, 1H), 7.20-1.08 (m, 1H), 6.63 (s, 1H), 4.90-4.73 (m, 1H), 4.60-3.80 (m, 9H), 3.75-3.40 (m, 9H), 3.30-2.80 (m, 6H), 2.40- 2.32 (m, 1H), 2.30-2.15 (m, 1H), 1.23 (brs, 3H); MS (LCMS) 727.4 [M+H]+. The absolute stereochemistry of Example 5A was arbitrarily assigned. Example 5B (Sa)-(-)-(Z)-16-Chloro-25-ethyl-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyrazina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0163] Example 5
Figure imgf000082_0002
mediate 73B following a similar procedure for the preparation of Example 1. Example 5B: (212 mg, 77%), off-white solid; 99.2% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.35 (br s, 1H), 11.50-10.91 (m, 1H), 8.15-8.08 (m, 1H), 7.92-7.80 (brs, 1H), 7.73-7.70 (m, 1H), 7.54-7.43 (m, 2H), 7.40 (s, 1H), 7.20-1.08 (m, 1H), 6.64 (s, 1H), 4.90-4.70 (m, 1H), 4.60-3.80 (m, 8H), 3.74 (s, 3H), 3.65-3.40 (m, 7H), 3.30- 2.80 (m, 6H), 2.40-2.32 (m, 1H), 2.30-2.15 (m, 1H), 1.23 (brs, 3H); MS (LCMS) 727.3 [M+H]+. The absolute stereochemistry of Example 5B was arbitrarily assigned. Intermediate 74 Ethyl (Z)-5-((tert-butyldiphenylsilyl)oxy)-2-hydroxy-4-oxopent-2-enoate [0164] t-BuOK (3.60 kg
Figure imgf000083_0001
d to THF (21 L), and the solution was cooled to 0 °C. Diethyl oxalate (4.69 kg, 32.1 mol) was added slowly, maintaining the temperature below 0 °C. The solution was stirred for 30 min at 0 °C. 1-((tert-Butyldiphenylsilyl)oxy)propan- 2-one (8.50 kg, 27.2 mol) was added slowly, maintaining the temperature below 0 °C. The mixture was stirred at 0 °C for 1 h. Upon completion by TLC, the reaction was diluted with EtOAc (5 L). The mixture was acidified with 1 N HCI to pH~2 to 3. The phases were separated, and the aqueous phase was extracted with EtOAc (8 L, 3 L). The combined organic phases were washed with brine, dried (Na2SO4), filtered and concentrated to afford Intermediate 74 (12.4 kg, crude) as an oil. Intermediate 75 Ethyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazole-3-carboxylate [0165] Intermediate 74
Figure imgf000083_0002
mol) was dissolved in 1,1,1,3,3,3- hexafluoropropan-2-ol (3.60 L) and trifluoroethanol (3.60 L). Two reactions of equal size were run simultaneously. The solution was cooled to 0 °C. Methylhydrazine (2.01 kg, 17.5 mol) was added dropwise at 0 °C. The ice bath was removed, and the mixture was stirred at rt for 2 h. Upon completion by TLC, the reactions were combined and concentrated. Water (7 L) was added, and the mixture was extracted with EtOAc (5 L, 3 L, 2 L). The organic layers were washed with brine (3 L), dried (Na2SO4) and filtered. The solvent was then removed. The residue was purified by flash chromatography (SiO2, EtOAc/Pet. Ether) to afford Intermediate 75 (3.50 kg, 24%, 2-steps) as an oil. 1H NMR (400 MHz, CDCl3) δ 7.64-7.67 (m, 4H), 7.39-7.49 (m, 6H), 6.56 (s, 1H), 4.68 (s, 2H), 4.38-4.43 (m, 2H), 3.95 (s, 3H), 1.41 (t, J=7.0 Hz, 3H), 1.05 (s, 9H). Intermediate 76 Ethyl 5-((acetylthio)methyl)-1-methyl-1H-pyrazole-3-carboxylate [0166] Intermediate 75 (3
Figure imgf000083_0003
l) was dissolved in THF (7 L) at rt. 1 M TBAF (8.28 L) was added, and the reaction was stirred at rt for 1 h. Upon completion by TLC, the solvent was removed under reduced pressure. Brine (10 L) was added to the residue. The mixture was extracted with EtOAc (10 L, 1 L x 10). The combined organic phases were dried (Na2SO4), filtered and concentrated. The residue was purified by flash chromatography (SiO2, EtOAc/Pet. ether) to afford ethyl 5-(hydroxymethyl)-1-methyl-1H-pyrazole-3-carboxylate (1.30 kg, 82%) as an oil. [0167] A solution of compound ethyl 5-(hydroxymethyl)-1-methyl-1H-pyrazole-3- carboxylate (1.30 kg) in DCM (7.80 L) was cooled to 0 oC. SOCl2 (924 g, 7.76 mol) was added. The ice bath was removed, and the reaction was stirred at rt for 1 h. Upon completion by TLC, the mixture was concentrated to dryness. EtOAc (1.5 L) was added to the residue. The solution was washed with sat. NaHCO3 (2 x 500 mL), dried (Na2SO4), filtered and concentrated to afford ethyl 5-(chloromethyl)-1-methyl-1H-pyrazole-3-carboxylate (1.28 kg, 89.5%) as an oil. [0168] To a mixture of compound ethyl 5-(chloromethyl)-1-methyl-1H-pyrazole-3- carboxylate (1.28 kg, 6.32 mol) in CH3CN (7.20 L) was added KI (1.05 kg, 6.32 mol) in one portion at rt under N2. The mixture was stirred at rt for 15 min, then AcSK (1.08 kg, 9.48 mol) was added. The reaction was then stirred at 60 °C for 1 h where the reaction was determined to be complete by TLC. The mixture was concentrated to dryness. Water (5 L) and EtOAc (4 L) was added to the residue. The layers were separated. The organic phase was dried (Na2SO4) and concentrated. The residue was purified by flash chromatography (SiO2, EtOAc) to afford Intermediate 76 (1.36 kg, 85%) as a brown solid. MS (LCMS) 243.0 [M+H]+. Intermediate 77 Ethyl 4-(3-fluorophenyl)-3-oxobutanoate [0169] To a stirred soluti
Figure imgf000084_0001
yl)acetic acid (250 g, 1.62 mol) in DCM (1.25 L) was added oxalyl chloride (166 mL, 1.95 mol) at 0oC. DMF (5 mL) was then added dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 4 h. The mixture was concentrated to afford 2-(3-fluorophenyl)acetyl chloride 280 g (crude) as a pale yellow liquid. [0170] To a stirred solution of Meldrum’s acid (222 g, 1.54 mol) in DCM (2.8 L) was added pyridine (327 mL, 4.04 mol) at 0 °C. The reaction was stirred at 0 oC for 10 min. 2-(3- Fluorophenyl)acetyl chloride (280 g, 1.62 mol) was added dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was quenched with 2N HCl (1 L) at 0 °C. The layers were separated, and the aqueous phase was extracted with DCM (2 x 1 L). The combined organic layers were dried (Na2SO4), filtered and concentrated. The brown solid (320 g) was dissolved in EtOH (960 mL) and heated to 90 °C for 4 h. Upon completion, the mixture was concentrated. The crude material was purified by flash chromatography (SiO2, 10% EtOAc/Pet. ether) to afford Intermediate 77 (200 g, 62% 2-steps) as an oil. MS (LCMS) 225.3 [M+H]+. Intermediate 78 6-Fluoronaphthalene-1,3-diol [0171] Intermediate 77 (5
Figure imgf000085_0001
ol) was dissolved in H2SO4 (150 mL) and stirred at rt for 16 h. Upon completion, the reaction was poured into ice cold water (500 mL), and the mixture was made alkaline by slowly adding sat. NaHCO3 (2 L). The mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (1 L), dried (Na2SO4) and concentrated. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 78 (25 g, 78%) as a yellow solid. MS (LCMS) 179.4 [M+H]+. Intermediate 79 4-((tert-Butyldimethylsilyl)oxy)-7-fluoronaphthalen-2-ol [0172] To a stirred solution
Figure imgf000085_0002
te 78 (30 g, 168 mmol) in THF (250 mL) was added imidazole (23 g, 338 mmol) followed by TBSCl (30.5 g, 202 mmol) in THF (50 mL) at 0°C. The ice bath was removed, and the reaction was stirred at rt for 16 h. The reaction was quenched with ice water (300 mL) and extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (1 L), dried (Na2SO4), filtered and concentrated. The reaction was run six times on an equivalent scale. The reactions were combined for purification. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 79 (35 g, 12%) as an oil. 1H NMR (400 MHz, DMSO-d6) δ 9.83 (s, 1H), 7.98 (dd, J=8.8, 2.8 Hz, 1H), 7.44 (dd, J=10.8, 2.4, 1H), 7.17-7.12 (m, 1H), 6.78 (d, J=2.0 Hz, 1H), 6.55 (d, J=2.0 Hz, 1H), 1.07 (s, 9H), 0.28 (s, 6H); MS (LCMS) 293.1 [M+H]+. 3-((tert- Butyldimethylsilyl)oxy)-6-fluoro-naphthalen-1-ol and ((6-fluoronaphthalene-1,3- diyl)bis(oxy))bis(tert-butyldimethylsilane) were also observed. Intermediate 80 4-((tert-Butyldimethylsilyl)oxy)-7-fluoronaphthalen-2-yl trifluoromethanesulfonate [0173] To a stirred solution
Figure imgf000086_0001
te 79 (25 g, 85.5 mmol) in DCM (250 mL) and DIPEA (25.6 mL, 147 mmol) was added trifluoromethanesulfonic anhydride (15.1 mL, 90.0 mmol) dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 1 h. Upon completion by TLC, the reaction was quenched with cold water (250 mL). The layers were separated, and the aqueous layer was extracted with DCM (3 x 150 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated. The crude material was purified by flash chromatography (SiO2, 15% EtOAc/Pet. ether) to afford Intermediate 80 (23 g, 63%) as an oil. 1H NMR (400 MHz, CDCl3) δ 8.16 (dd, J=9.2, 5.6 Hz, 1H), 7.41 (dd, J=9.6, 2.4 Hz, 1H), 7.29- 7.25 (m, 2H), 6.68 (d, J=2.4 Hz, 1H), 1.07 (s, 9H), 0.30 (s, 6H). (LCMS) 423.3 [M-H]-. Intermediate 81 Ethyl 5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H-pyrazole-3-carboxylate [0174] To a degasse
Figure imgf000086_0002
(44.0 g, 103.7 mmol) in 1,4-dioxane (880 mL):water (44 mL) was added Intermediate 76 (22.6 g, 93.3 mmol), K2CO3 (15.0 g, 109 mmol), XPhos (4.93 g, 10.3 mmol) and Pd2(dba)3 (4.75 g, 5.19 mmol). The solution was degassed with Ar for 15 min. The reaction was heated at 100°C for 3 h. Upon completion, the reaction was cooled to rt, diluted with EtOAc (250 mL) and passed through a Celite pad. The Celite pad was washed with EtOAc (200 mL). The organic layer was washed with brine (500 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 15% EtOAc/Pet. ether) to afford Intermediate 81 (25 g, 67%) as an off-white solid. MS (LCMS) 361.3 [M+H]+. Intermediate 82 6-Fluoro-3-(((3-(hydroxymethyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-ol [0175] To a stirred sol
Figure imgf000087_0001
30.0 g, 83.2 mmol) in THF (300 mL) was added 2.4 M LiAlH4 in THF (52.0 mL, 125 mmol) dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction was cooled to 0 °C and quenched with sat. Na2SO4 (25 mL). The precipitate was removed by filtration, and the filtrate was dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 82 (17 g, 64%) as an off-white solid. MS (LCMS) 319.3 [M+H]+. Intermediate 83 3-(((3-(Chloromethyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)-6-fluoronaphthalen-1-ol OH [ 0 mL) under A
Figure imgf000087_0003
r was a e . g, mmo , , - u ne . m , mmo and MsCl (4.20 mL, 50.3 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction was quenched with water (300 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (300 mL), dried (Na2SO4), filtered and concentrated to afford Intermediate 83 (8.5 g) as an oil. (LCMS) 337.3 [M+H]+. Intermediate 84 3-(((3-(Chloromethyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)-6-fluoronaphthalen-1-yl acetate OAc [0177 mL) was add
Figure imgf000087_0002
was stirred at rt for 2 h. Upon completion by TLC, the reaction was quenched with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (50 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 84 (10 g) as an oil. MS (LCMS) 379.2 [M+H]+. Intermediate 85 3-(((3-((Acetylthio)methyl)-1-methyl-1H-pyrazol-5-yl)methyl)thio)-6-fluoronaphthalen-1-yl acetate [0178] To a degassed
Figure imgf000088_0001
(10.0 g, 26.5 mmol) in CH3CN (100 mL) was added KSAc (4.40 g, 26.5 mmol) and KI (6.00 g, 52.9 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was quenched with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (200 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 85 (4.7 g, 45%, 3-steps) as an off-white solid. MS (LCMS) 419.3 [M+H]+. Intermediate 86 Methyl 6-chloro-7-(1-ethyl-5-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl- 1H-pyrazol-3-yl)methyl)thio)methyl)-3-methyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1-methyl- 1H-indole-2-carboxylate OH
Figure imgf000088_0002
Intermediate 87A (Ra)-(+)-Methyl (Z)-16-chloro-21-ethyl-96-fluoro-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8- dithia-1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12- carboxylate (Sa)-(-)-Methyl (Z)-16
Figure imgf000089_0001
-chloro-21-ethyl-96-fluoro-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8- dithia-1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12- carboxylate [0180] Intermediat
Figure imgf000089_0002
ed from Intermediate 86 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-16-chloro-21-ethyl-96-fluoro-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (400 mg, 41%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 25% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Intermediate 87A, 190 mg) and peak 2 (Intermediate 87B, 170 mg). Intermediate 87A: off-white solid; 99.9% chiral purity; MS (LCMS) 718.2 [M+H]+. Intermediate 87B: off-white solid; 99.3% chiral purity; MS (LCMS) 718.2 [M+H]+. The absolute stereochemistry of Intermediate 87A and Intermediate 87B was arbitrarily assigned. Example 6A (Ra)-(+)-(Z)-16-Chloro-21-ethyl-96-fluoro-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0181] Example 6A
Figure imgf000090_0001
mediate 87A following a similar procedure for the preparation of Example 1. Example 6A: (150 mg, 85%), white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.3 (br s, 1H), 8.19 (dd, J=9.2, 6.0 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 7.51 (dd, J=10.4, 2.0 Hz, 1H), 7.36-7.30 (m, 2H), 7.01 (d, J=8.8 Hz, 1H), 6.65 (s, 1H), 4.85 (s, 1H), 4.40-4.30 (m, 1H), 4.22-3.95 (m, 4H), 3.93-3.85 (m, 1H), 3.69 (s, 3H), 3.56 (d, J=14.0 Hz, 1H), 3.45-3.40 (m, 4H), 3.30-3.20 (m, 1H), 3.13 (d, J=13.6 Hz, 1H), 3.05-2.95 (m, 2H), 2.35-2.20 (m, 2H), 1.88 (s, 3H), 1.38 (t, J=7.2 Hz, 3H); MS (LCMS) 704.6 [M+H]+. The absolute stereochemistry of Example 6A was arbitrarily assigned. Example 6B (Sa)-(-)-(Z)-16-Chloro-21-ethyl-96-fluoro-11,23,61-trimethyl-11H,21H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0182] Example 6B
Figure imgf000090_0002
mediate 87B following a similar procedure for the preparation of Example 1. Example 6B: (65 mg, 40%), white solid; 99.4% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.3 (br s, 1H), 8.19 (dd, J=9.2, 6.0 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 7.53 (dd, J=10.8, 2.0 Hz, 1H), 7.38-7.32 (m, 2H), 7.06 (d, J=8.8 Hz, 1H), 6.59 (s, 1H), 4.74 (s, 1H), 4.30-4.20 (m, 2H), 4.25-3.95 (m, 3H), 3.90-3.80 (m, 1H), 3.69 (s, 3H), 3.55- 3.40 (m, 5H), 3.25-3.20 (m, 1H), 3.18-3.05 (m, 2H), 2.94 (d, J=14.0 Hz, 1H), 2.40-2.32 (m, 1H), 2.30-2.20 (m, 1H), 1.89 (s, 3H), 1.34 (t, J=7.2 Hz, 3H); MS (LCMS) 704.6 [M+H]+. The absolute stereochemistry of Example 6B was arbitrarily assigned. Intermediate 88 Methyl 6-fluoro-7-(5-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H- pyrazol-3-yl)methyl)thio)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1-methyl- 1H-indole-2-carboxylate [0183] Intermedi
Figure imgf000091_0001
mediate 35 and Intermediate 85 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 706.6 [M+H]+. Intermediate 89A (Ra)-(+)-Methyl (Z)-16,96-difluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (Sa)-(-)-Methyl (Z)-16,96
Figure imgf000091_0002
-difluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0184] Intermediate
Figure imgf000091_0003
ed from Intermediate 88 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-16,96-difluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (170 mg, 37%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 30% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Intermediate 89A, 60 mg) and peak 2 (Intermediate 89B, 60 mg). Intermediate 89A: off-white solid; 99.6% chiral purity; MS (LCMS) 688.7 [M+H]+. Intermediate 89B: off-white solid; 98.6% chiral purity; MS (LCMS) 688.7 [M+H]+. The absolute stereochemistry of Intermediate 89A and Intermediate 89B was arbitrarily assigned. Example 7A (Ra)-(+)-(Z)-16,96-Difluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0185] Example 7A
Figure imgf000092_0001
rmediate 89A following a similar procedure for the preparation of Example 1. Example 7A: (32 mg, 56%), white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.2 (br s, 1H), 8.18 (dd, J=9.2, 3.6 Hz, 1H), 7.77 (dd, J=8.8, 5.6 Hz, 1H), 7.52 (dd, J=10.4, 2.4 Hz, 1H), 7.37-7.32 (m, 2H), 6.80 (t, J=9.2 Hz, 1H), 6.65 (s, 1H), 4.71 (s, 1H), 4.31-4.19 (m, 2H), 4.07-4.01 (m, 1H), 3.87-3.86 (m, 1H), 3.76 (s, 3H), 3.69 (s, 3H), 3.52 (s, 3H), 3.49-3.47 (m, 2H), 3.44-3.31 (m, 1H), 3.15-3.11 (m, 2H), 2.98 (d, J=14.4 Hz, 1H), 2.49-2.40 (m, 1H), 2.37-2.24 (m, 1H), 1.90 (s, 3H); MS (LCMS) 674.6 [M+H]+. The absolute stereochemistry of Example 7A was arbitrarily assigned. Example 7B (Sa)-(-)-(Z)-16,96-Difluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid
Figure imgf000092_0002
[0186] Example 7B was synthesized from Intermediate 89B following a similar procedure for the preparation of Example 1. Example 7B: (36 mg, 61%), white solid; 98.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.2 (br s, 1H), 8.18 (dd, J=9.2, 6.0 Hz, 1H), 7.77 (dd, J=8.8, 5.6 Hz, 1H), 7.52 (dd, J=10.0, 2.0 Hz, 1H), 7.37-7.32 (m, 2H), 6.80 (t, J=9.2 Hz, 1H), 6.65 (s, 1H), 4.72 (s, 1H), 4.31-4.19 (m, 2H), 4.07-4.01 (m, 1H), 3.88-3.86 (m, 1H), 3.76 (s, 3H), 3.69 (s, 3H), 3.52 (s, 3H), 3.49-3.43 (m, 2H), 3.32 (d, J=14.4 Hz, 1H), 3.13 (d, J=14.4, 2H), 2.98 (d, J=14.0 Hz, 1H), 2.38-2.37 (m, 1H), 2.32-2.24 (m, 1H), 1.90 (s, 3H); MS (LCMS) 674.6 [M+H]+. The absolute stereochemistry of Example 7B was arbitrarily assigned. Intermediate 90 Methyl 7-(5-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H-pyrazol-3- yl)methyl)thio)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3-hydroxypropyl)-1,6-dimethyl-1H- indole-2-carboxylate OH
Figure imgf000093_0002
following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 702.8 [M+H] . Intermediate 91A (Ra)-(+)-Methyl (Z)-96-fluoro-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000093_0001
Intermediate 91B (Sa)-(-)-Methyl (Z)-96-fluoro-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0188] Intermediate
Figure imgf000094_0001
zed from Intermediate 90 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-96-fluoro-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (720 mg, 73%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OJ-3 (30 x 250 mm), 30% MeOH) to give peak 1 (Intermediate 91A, 60 mg) and peak 2 (Intermediate 91B, 30 mg). Intermediate 91A: off-white solid; 99.9% chiral purity; MS (LCMS) 684.7 [M+H]+. Intermediate 91B: off-white solid; 99.4% chiral purity; MS (LCMS) 684.9 [M+H]+. The absolute stereochemistry of Intermediate 91A and Intermediate 91B was arbitrarily assigned. Example 8A (Ra)-(+)-(Z)-96-Fluoro-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0189] Example 8A
Figure imgf000094_0002
rmediate 91A following a similar procedure for the preparation of Example 1. Example 8A: (35 mg, 60%), white solid; 98.5% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.0 (s, 1H), 8.21 (dd, J=8.8, 5.6 Hz, 1H), 7.59- 7.56 (m, 2H), 7.41-7.35 (m, 2H), 6.86 (d, J=8.4 Hz, 1H), 6.52 (s, 1H), 4.73 (s, 1H), 4.28 ( d, J=15.6 Hz, 1H), 4.12 (d, J=15.2 Hz, 1H), 3.92-3.86 (m, 2H), 3.75 (s, 3H), 3.65 (s, 3H), 3.52-3.40 (m, 5H), 3.19-3.03 (m, 3H), 2.87 (d, J=14.0 Hz, 1H), 2.36-2.18 (m, 2H), 1.95 (s, 3H), 1.86 (s, 3H); MS (LCMS) 670.6 [M+H]+. The absolute stereochemistry of Example 8A was arbitrarily assigned. Example 8B (Sa)-(-)-(Z)-96-Fluoro-11,16,21,23,61-pentamethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0190] Example 8B
Figure imgf000095_0001
rmediate 91B following a similar procedure for the preparation of Example 1. Example 8B: (20 mg, 69%), white solid; 99.2% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.0 (s, 1H), 8.21 (dd, J= 8.4, 2.4 Hz, 1H), 7.59- 7.54 (m, 2H), 7.41-7.35 (m, 2H), 6.85 (d, J=8.4 Hz, 1H), 6.52 (s, 1H), 4.73 (s, 1H), 4.28 ( d, J=16.0 Hz, 1H), 4.12 (d, J=15.6 Hz, 1H), 3.92-3.88 (m, 2H), 3.74 (s, 3H), 3.68 (s, 3H), 3.52-3.31 (m, 5H), 3.18-3.06 (m, 3H), 2.87 (d, J=13.6 Hz, 1H), 2.32-2.18 (m, 2H), 1.95 (s, 3H), 1.86 (s, 3H); MS (LCMS) 670.6 [M+H]+. The absolute stereochemistry of Example 8B was arbitrarily assigned. Intermediate 92 Methyl 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-2-carboxylate N N O O [0191] To a stirred solution
Figure imgf000095_0002
dro-4H-pyrrolo[1,2-b]pyrazole-2-carboxylic acid (10.0 g, 65.7 mmol) in MeOH (100 mL) was added SOCl2 (15.64 g, 131.4 mmol) at 0 °C. The reaction was stirred at reflux for 6 h. Upon completion by TLC, the solvent was evaporated and co-distilled with MeOH. The solid was dried under high vacuum to afford Intermediate 92 (10 g, 92%) as an off-white solid. MS (LCMS) 167.1 [M+H]+. Intermediate 93 (5,6-Dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)methanol [0192] To a stirred solution
Figure imgf000095_0003
te 92 (10.0 g, 60.2 mmol) in THF (100 ml) was added 2M LiAlH4 in THF (60.2 mL, 120 mmol) at 0 ^C. The reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction was quenched with sat. NH4Cl (100 mL) and extracted EtOAc (4 x 200 mL). The organic layers were combined, dried (Na2SO4) and filtered. The solvent was evaporated, and the residue was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 93 (6.5 g, 78%) as an off-white solid. MS (LCMS) 139.1 [M+H]+. Intermediate 94 (3-Bromo-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-2-yl)methanol [0193] To a stirred solution
Figure imgf000096_0001
te 93 (4.00 g, 28.9 mmol) in DCM (50 mL) was added NBS (5.18 g, 28.9 mmol) at 0 °C. The reaction was stirred at rt for 2 h. Upon completion by TLC, the mixture was diluted with sat. NaHCO3 (50 mL) and extracted with DCM (3 x 50 mL). The organic layers were combined, dried (Na2SO4) and filtered. The solvent was evaporated, and the residue was triturated with pentane:ether (1:1) (3 x 20 mL) to provide Intermediate 94 (5.0 g, 79%) as a yellow solid. MS (LCMS) 217.0 [M+H]+. Intermediate 95 3-Bromo-2-(((4-methoxybenzyl)oxy)methyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole [0194] To a stirred solution
Figure imgf000096_0002
e 94 (4.00 g, 18.4 mmol) in DMF (40 mL) was added 60% NaH (1.1 g, 27.6 mmol) at 0 °C. The reaction was stirred at rt for 30 min. 1- (Chloromethyl)-4-methoxybenzene (4.04 g, 25.8 mmol) and KI (300 mg, 1.81 mmol) was added. The reaction was stirred at rt for 18 h. Upon completion, the reaction was quenched with sat. NH4Cl (50 ml). The mixture was extracted with EtOAc (4 x 50 mL). The combined organic layers were washed with water (2 x 50 mL), brine (50 mL) and dried (Na2SO4). The solvent was evaporated, and the residue was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 95 (3.4 g, 54%) as an off-white solid. MS (LCMS) 336.9 [M+H]+. Intermediate 96 2-(((4-Methoxybenzyl)oxy)methyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydro- 4H-pyrrolo[1,2-b]pyrazole [0195] To a stirred solution
Figure imgf000096_0003
e 95 (10.0 g, 29.8 mmol) in THF (200 mL) was added 1.6 M n-BuLi in hexanes (27.9 mL, 44.6 mmol) at -78 °C. The reaction was stirred at -78 °C for 50 min. 2-Isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (22.14 mL, 119.0 mmol) was added at -78 °C, and the reaction was stirred at -78 °C for 1 h. Upon completion, the reaction temperature was gradually increased to rt. The solvent was removed by evaporation under reduced pressure, and the reaction was diluted with EtOAc (200 mL). The mixture was filtered through a Celite pad and washed with EtOAc (2 x 50 mL). The solvent was evaporated, and the residue was purified by flash chromatography (SiO2, 30-50% EtOAc/Pet. ether) to afford Intermediate 96 (8.8 g, 88%) as a white solid. MS (LCMS) 385.4 [M+H]+.
Intermediate 97
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(((4-methoxybenzyl)oxy)methyl)-5,6-dihydro-4H- pyrrolo[l,2-b]pyrazol-3-yl)-l-methyl-lH-indole-2-carboxylate
Figure imgf000097_0001
[0196] Intermediate 97 was synthesized from Intermediate 4 and Intermediate 96 following a similar procedure for the preparation of Intermediate 20. MS (LCMS) 580.5 [M+H]+.
Intermediate 98
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(hydroxymethyl)-5,6-dihydro-4H-pyrrolo[l,2- b]pyrazol-3-yl)-l-methyl-lH-indole-2-carboxylate
Figure imgf000097_0002
[0197] Intermediate 98 was synthesized from Intermediate 97 following a similar procedure for the preparation of Intermediate 21. MS (LCMS) 460.8 [M+H]+. Intermediate 99
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(chloromethyl)-5,6-dihydro-4H-pyrrolo[l,2- b]pyrazol-3-yl)- 1-methyl- lH-indole-2-carboxylate
Figure imgf000098_0001
[0198] Intermediate 99 was synthesized from Intermediate 98 following a similar procedure for the preparation of Intermediate 22. MS (LCMS) 478.4 [M+H]+.
Intermediate 100
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(iodomethyl)-5,6-dihydro-4H-pyrrolo[l,2-b]pyrazol- 3 -yl)- 1-methyl- lH-indole-2-carboxylate
Figure imgf000098_0002
[0199] Intermediate 100 was synthesized from Intermediate 99 following a similar procedure for the preparation of Intermediate 23. MS (LCMS) 570.4 M+H]+.
Intermediate 101
Methyl 6-chloro-7-(2-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)- 1-methyl- 1H- pyrazol-3-yl)methyl)thio)methyl)-5,6-dihydro-4H-pyrrolo[l,2-b]pyrazol-3-yl)-3-(3- hydroxypropyl)- 1 -methyl- 1 H-indole-2-carboxylate
Figure imgf000098_0003
[0200] Intermediate 101 was synthesized from Intermediate 100 and Intermediate
85 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 734.6 [M+H]+. Intermediate 102A (Ra)-(+)-Methyl (Z)-16-chloro-96-fluoro-11,61-dimethyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8- dithia-1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (Sa)-(-)-Methyl (Z)-16
Figure imgf000099_0001
-chloro-96-fluoro-11,61-dimethyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8- dithia-1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate [0201] Intermedia
Figure imgf000099_0002
thesized from Intermediate 101 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-16-chloro-96-fluoro-11,61-dimethyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (360 mg, 39%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 25% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Intermediate 102A, 110 mg) and peak 2 (Intermediate 102B, 110 mg). Intermediate 102A: off-white solid; 99.3% chiral purity; MS (LCMS) 716.3 [M+H]+. Intermediate 102B: off-white solid; 99.8% chiral purity; MS (LCMS) 716.2 [M+H]+. The absolute stereochemistry of Intermediate 102A and Intermediate 102B was arbitrarily assigned. Example 9A (Ra)-(+)-(Z)-16-Chloro-96-fluoro-11,61-dimethyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0202] Example 9
Figure imgf000100_0001
mediate 102A following a similar procedure for the preparation of Example 1. Example 9A: (75 mg, 65%), white solid; 99.5% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.3 (br s, 1H), 8.14 (dd, J=9.2, 6.0 Hz, 1H), 7.85 (d, J=8.8 Hz, 1H), 7.50 (dd, J=10.4, 2.4 Hz, 1H), 7.38-7.30 (m, 2H), 7.13 (d, J=8.8 Hz, 1H), 6.67 (s, 1H), 4.78 (s, 1H), 4.29 (s, 2H), 4.18-4.08 (m, 3H), 3.90-3.82 (m, 1H), 3.71 (s, 3H), 3.54 (s, 3H), 3.50-3.40 (m, 2H), 3.24 (d, J=12.8 Hz, 1H), 3.15-3.05 (m, 2H), 2.92 (d, J=14.0 Hz, 1H), 2.70-2.50 (m, 4H), 2.42-2.30 (m, 1H), 2.28-2.15 (m, 1H); MS (LCMS) 702.2 [M+H]+. The absolute stereochemistry of Example 9A was arbitrarily assigned. Example 9B (Sa)-(-)-(Z)-16-Chloro-96-fluoro-11,61-dimethyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0203] Example 9B
Figure imgf000100_0002
mediate 102B following a similar procedure for the preparation of Example 1. Example 9B: (82 m, 65%), white solid; 99.4% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.3 (br s, 1H), 8.14 (dd, J=9.2, 6.0 Hz, 1H), 7.85 (d, J=8.8 Hz, 1H), 7.52 (dd, J=10.4, 2.4 Hz, 1H), 7.38-7.30 (m, 2H), 7.13 (d, J=8.8 Hz, 1H), 6.67 (s, 1H), 4.78 (s, 1H), 4.29 (s, 2H), 4.18-4.08 (m, 3H), 3.90-3.82 (m, 1H), 3.71 (s, 3H), 3.54 (s, 3H), 3.48-3.40 (m, 2H), 3.24 (d, J=12.8 Hz, 1H), 3.15-3.05 (m, 2H), 2.92 (d, J=14.4 Hz, 1H), 2.70-2.50 (m, 4H), 2.42-2.30 (m, 1H), 2.28-2.15 (m, 1H); MS (LCMS) 702.0 [M+H]+. The absolute stereochemistry of Example 9B was arbitrarily assigned.
Intermediate 103
2-(((4-Methoxybenzyl)oxy)methyl)-3-(4, 4,5, 5 -tetramethyl- 1,3, 2-dioxaborolan-2-yl)-4, 5,6,7- tetrahy dropyrazolo [ 1 , 5 - a] pyridine
Figure imgf000101_0001
[0204] Intermediate 103 was synthesized from 4,5,6,7-tetrahydropyrazolo[l,5- a]pyridine-2-carboxylic acid following a similar procedure for the preparation of Intermediates 92-96. MS (LCMS) 399.4 [M+H]+.
Intermediate 104
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(((4-methoxybenzyl)oxy)methyl)-4,5,6,7- tetrahydropyrazolo[l ,5-a]pyridin-3-yl)- 1 -methyl- lH-indole-2-carboxylate
Figure imgf000101_0002
[0205] Intermediate 104 was synthesized from Intermediate 4 and Intermediate 103 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 594.4 [M+H]+.
Intermediate 105
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(hydroxymethyl)-4,5,6,7-tetrahydropyrazolo[l,5- a]pyridin-3-yl)-l-methyl-lH-indole-2-carboxylate
Figure imgf000101_0003
[0206] Intermediate 105 was synthesized from Intermediate 104 following a similar procedure for the preparation of Intermediate 21. MS (LCMS) 474.4 [M+H]+. Intermediate 106
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(chloromethyl)-4,5,6,7-tetrahydropyrazolo[l,5- a]pyridin-3-yl)-l-methyl-lH-indole-2-carboxylate
Figure imgf000102_0001
[0207] Intermediate 106 was synthesized from Intermediate 105 following a similar procedure for the preparation of Intermediate 22. MS (LCMS) 492.4 [M+H]+.
Intermediate 107
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(2-(iodomethyl)-4,5,6,7-tetrahydropyrazolo[l,5- a]pyridin-3-yl)-l-methyl-lH-indole-2-carboxylate
Figure imgf000102_0002
[0208] Intermediate 107 was synthesized from Intermediate 106 following a similar procedure for the preparation of Intermediate 23. MS (LCMS) 584.2 [M+H]+.
Intermediate 108
Methyl 6-chloro-7-(2-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-l-methyl-lH- pyrazol-3-yl)methyl)thio)methyl)-4,5,6,7-tetrahydropyrazolo[l,5-a]pyridin-3-yl)-3-(3- hydroxypropyl)- 1 -methyl- 1 H-indole-2-carboxylate
Figure imgf000102_0003
[0209] Intermediate 108 was synthesized from Intermediate 107and Intermediate
85 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 748.7 [M+H]+. Intermediate 109A (Ra)-(+)-Methyl (Z)-16-chloro-96-fluoro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa- 4,8-dithia-2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (Sa)-(-)-Methyl (Z)-16
Figure imgf000103_0001
-chloro-96-fluoro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa- 4,8-dithia-2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate [0210] Intermedia
Figure imgf000103_0002
thesized from Intermediate 108 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-16-chloro-96-fluoro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (450 mg, 46%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OX-H (30 x 250 mm), 40% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Intermediate 109A, 85 mg) and peak 2 (Intermediate 109B, 140 mg). Intermediate 109A: off-white solid; 98.6% chiral purity; MS (LCMS) 730.5 [M+H]+. Intermediate 109B: off-white solid; 98.7% chiral purity; MS (LCMS) 730.6 [M+H]+. The absolute stereochemistry of Intermediate 109A and Intermediate 109B was arbitrarily assigned. Example 10A (Ra)-(+)-(Z)-16-Chloro-96-fluoro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8- dithia-2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0211] Example 10
Figure imgf000104_0001
mediate 109A following a similar procedure for the preparation of Example 1. Example 10A: (41 mg, 49%), off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.3 (s, 1H), 8.19-8.11 (m, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.51 (d, J=9.2 Hz, 1H), 7.35-7.31 (m, 2H), 7.14 (d, J=8.4 Hz, 1H), 6.67 (s, 1H), 4.77 (s, 1H), 4.29 (s, 2H), 4.14-4.07 (m, 3H), 3.86-3.85 (m, 1H), 3.72 (s, 3H), 3.51 (s, 3H), 3.46-3.42 (m, 2H), 3.19-3.07 (m, 3H), 2.95-2.89 (m, 1H), 2.39-2.24 (m, 4H), 1.97-1.74 (m, 4H); MS (LCMS) 716.6 [M+H]+. The absolute stereochemistry of Example 10A was arbitrarily assigned. Example 10B (Sa)-(-)-(Z)-16-Chloro-96-fluoro-11,61-dimethyl-24,25,26,27-tetrahydro-11H,61H-10-oxa-4,8-dithia- 2(3,2)-pyrazolo[1,5-a]pyridina-1(7,3)-indola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0212] Example 10
Figure imgf000104_0002
mediate 109B following a similar procedure for the preparation of Example 1. Example 10B: (74 mg, 54%), off-white solid; 98.5% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.3 (s, 1H), 8.15-8.12 (m, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.51 (d, J=10.0 Hz, 1H), 7.35-7.31 (m, 2H), 7.14 (d, J=8.4 Hz, 1H), 6.66 (s, 1H), 4.74 (s, 1H), 4.28 (s, 2H), 4.14-4.06 (m, 3H), 3.85-3.84 (m, 1H), 3.71 (s, 3H), 3.51 (s, 3H), 3.45-3.42 (m, 2H), 3.22-3.05 (m, 3H), 2.95-2.91 (m, 1H), 2.39-2.23 (m, 4H), 1.96-1.72 (m, 4H); MS (LCMS) 716.4 [M+H]+. The absolute stereochemistry of Example 10B was arbitrarily assigned. Intermediate 110 Ethyl 3-(((tert-butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazole-5-carboxylate [0213] To a stirred soluti
Figure imgf000105_0001
5 (47.0 g, 115 mmol) in DMF (450 mL) was added Cs2CO3 (112 g, 345 mmol) and MeI (14.3 mL, 230 mmol) at 0°C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by LCMS, water (800 mL) was added. The mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with water (800 mL), brine (1 L), dried (Na2SO4) filtered and concentrated. The crude residue was purified by flash chromatography (SiO2, EtOAc/Pet. ether) to afford ethyl 5- (((tert-butyldiphenylsilyl)oxy)methyl)-1-methyl-1H-pyrazole-3-carboxylate (18 g, 36%) and Intermediate 110 (31 g, 63%) as an oil. 1H NMR (400 MHz, CDCl3) δ 7.71-7.67 (m, 4H), 7.45- 7.35 (m, 6H), 6.80 (s, 1H), 4.72 (s, 2H), 4.35 (q, J=7.2 Hz, 2H), 4.11 (s, 3H), 1.39 (t, J=7.2 Hz, 3H), 1.08 (s, 9H). MS (LCMS) 423.4 [M+H]+. Intermediate 111 Ethyl 3-(chloromethyl)-1-methyl-1H-pyrazole-5-carboxylate [0214] To a stirred solution
Figure imgf000105_0002
e 110 (31.0 g, 73.3 mmol) in THF (310 mL) was added 1M TBAF in THF (73.3 mL) and CsF (22.8 g, 147 mmol) at 0 °C. The reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was diluted with EtOAc (300 mL) and washed with water (300 mL). The organic layer was dried (Na2SO4), filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography (SiO2, EtOAc/Pet. ether) to afford ethyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-5-carboxylate (9 g, 66%) as a yellow solid. MS (LCMS) 185.2 [M+H]+. [0215] To a stirred solution of ethyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-5- carboxylate (9.00 g, 48.9 mmol) in DCM (90 mL) under Ar was added SOCl2 (12.0 mL, 174 mmol) at 0 °C. The reaction was stirred at rt for 2 h. The solvent was evaporated and sat. NaHCO3 (150 mL) was added. The mixture was extracted with DCM (2 x 150 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated to afford Intermediate 111 (9.9 g). MS (LCMS) 203.1 [M+H]+. Intermediate 112 Ethyl 3-((acetylthio)methyl)-1-methyl-1H-pyrazole-5-carboxylate [0216] To a stirred solution
Figure imgf000106_0001
e 111 (9.88 g, 48.7 mmol) in CH3CN (100 mL) was added KI (8.10 g, 48.7 mmol) and KSAc (11.1 g, 97.3 mmol) at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 3 h. Upon completion by TLC, the reaction was concentrated and diluted with water (300 mL). The mixture was extracted with EtOAc (2 x 250 mL). The combined organic layers were washed with brine (500 mL), dried (Na2SO4), filtered and evaporated. The crude residue was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford Intermediate 112 (7.1 g, 60%, 2-steps) as a yellow solid. MS (LCMS) 243.1 [M+H]+. Intermediate 113 Ethyl 3-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-methyl-1H-pyrazole-5-carboxylate [0217] To a solution
Figure imgf000106_0002
g, 24.79 mmol) in 1,4-dioxane (120 mL) and water (30 mL) was added Intermediate 112 (6.00 g, 24.8 mmol), K2CO3 (3.76 g, 27.3 mmol) and XPhos (1.18 g, 2.48 mmol). The mixture was degassed under Ar for 15 min. Pd2(dba)3 (1.13 g, 1.24 mmol) was added, and the reaction was degassed for 15 min. The reaction was heated at 100°C for 3 h. The reaction was cooled to rt and filtered through a pad of Celite. The pad was washed with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (200 mL), dried (Na2SO4), filtered and evaporated. The crude residue was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford Intermediate 113 (5.3 g, 60%) as a yellow solid. MS (LCMS) 361.3 [M+H]+. Intermediate 114 3-(((5-(Chloromethyl)-1-methyl-1H-pyrazol-3-yl)methyl)thio)-6-fluoronaphthalen-1-yl acetate [0218] To a stirred sol
Figure imgf000107_0001
(5.30 g, 14.7 mmol) in THF (50 mL) was added 2.4 M LiAlH4 in THF (9.20 mL, 22.1 mol) dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 1 h. Upon completion by TLC, the reaction was cooled to 0 °C and quenched with sat. Na2SO4 (25 mL). The precipitated solid was removed by filtration through a bed of Celite. The bed was washed with EtOAc (75 mL). The filtrate was washed with brine (100 mL), dried (Na2SO4), filtered and evaporated to afford 6-fluoro-3-(((5- (hydroxymethyl)-1-methyl-1H-pyrazol-3-yl)methyl)thio)naphthalen-1-ol (3.5 g) as a brown solid. MS (LCMS) 319.3 [0219] To a stirred solution of 6-fluoro-3-(((5-(hydroxymethyl)-1-methyl-1H-pyrazol- 3-yl)methyl) thio)naphthalen-1-ol (5.00 g, 16.7 mmol) in DCM (50 mL) and DMF (5 mL) was added SOCl2 (1.5 mL, 20 mmol) dropwise. The reaction was stirred at rt for 3 h. After completion by TLC, the reaction was partitioned between sat. NaHCO3 (50 mL) and DCM (100 mL). The layers were separated, and the aqueous layer was extracted with DCM (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried (Na2SO4), filtered and evaporated to afford 3-(((5-(chloromethyl)-1-methyl-1H-pyrazol-3-yl)methyl)thio)-6- fluoronaphthalen-1-ol (3.5 g) as a brown solid. MS (LCMS) 337.2 [M+H]+. [0220] To a stirred solution of 3-(((5-(chloromethyl)-1-methyl-1H-pyrazol-3- yl)methyl)thio)-6-fluoronaphthalen-1-ol (3.50 g, 16.7 mmol) in CH3CN (35 mL) was added DMAP (202 mg, 1.67 mmol) and Ac2O (3.40 mL, 33.3 mmol) at 0 °C. The reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction was quenched with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (200 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 114 (4 g) as an oil. MS (LCMS) 379.2 [M+H]+. Intermediate 115
3-(((5-((Acetylthio)methyl)-l-methyl-lH-pyrazol-3-yl)methyl)thio)-6-fluoronaphthalen-l-yl acetate
Figure imgf000108_0001
[0221] To a degassed solution of Intermediate 114 (4.00 g, 10.6 mmol) in CH3CN (40 mL) was added KSAc (1.76 g, 10.6 mmol) and KI (1.21 g, 10.6 mmol) at 0 °C. The reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was quenched with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (200 mL), dried (Na2SO4), filtered and evaporated. The crude residue was purified by flash chromatography (SiCL, 20% EtOAc/Pet. ether) to afford Intermediate 115 (1.9 g, 30%, 4-steps) as an off-white solid. MS (LCMS) 419.3 [M+H]+.
Intermediate 116
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(5-(((4-methoxybenzyl)oxy)methyl)-l,3-dimethyl-lH- pyrazol-4-yl)- 1 -methyl- lH-indole-2-carboxylate
Figure imgf000108_0002
[0222] Intermediate 116 was synthesized from Intermediate 4 and Intermediate 29 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 568.0 [M+H]+.
Intermediate 117
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(5-(hydroxymethyl)-l,3-dimethyl-lH-pyrazol-4-yl)-l- methyl- lH-indole-2-carboxylate
Figure imgf000108_0003
[0223] Intermediate 117 was synthesized from Intermediate 116 following a similar procedure from the preparation of Intermediate 21. MS (LCMS) 448.5 [M+H]+.
Intermediate 118
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(5-(chloromethyl)-l,3-dimethyl-lH-pyrazol-4-yl)-l- methyl- lH-indole-2-carboxylate
Figure imgf000109_0001
[0224] Intermediate 118 was synthesized from Intermediate 117 following a similar procedure from the preparation of Intermediate 22. MS (LCMS) 465.9 [M+H]+.
Intermediate 119
Methyl 3-(3-acetoxypropyl)-6-chloro-7-(5-(iodomethyl)-l,3-dimethyl-lH-pyrazol-4-yl)-l- methyl- lH-indole-2-carboxylate
Figure imgf000109_0002
[0225] Intermediate 119 was synthesized from Intermediate 118 following a similar procedure from the preparation of Intermediate 23. MS (LCMS) 557.9 [M+H]+.
Intermediate 120
Methyl 6-chloro-7-(5-((((3-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-l-methyl-lH- pyrazol-5-yl)methyl)thio)methyl)-l,3-dimethyl-lH-pyrazol-4-yl)-3-(3-hydroxypropyl)-l-methyl- lH-indole-2-carboxylate
Figure imgf000109_0003
[0226] Intermediate 120 was synthesized from Intermediate 119 and Intermediate 115 following a similar procedure from the preparation of Intermediate 24. MS (LCMS) 722.1 [M+H]+. Intermediate 121A (Ra)-(+)-Methyl (Z)-16-chloro-96-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000110_0001
(Sa)-(-)-Methyl (Z)-16-chloro-96-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia- 1(7,3)-indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0227] Intermediates
Figure imgf000110_0002
hesized from Intermediate 120 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-16-chloro-96-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate (700 mg, 75%). The atropisomers were separated by chiral SFC chromatography (Lux Cellulose-2 (30 x 250 mm), 40% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Intermediate 121A, 120 mg) and peak 2 (Intermediate 121B, 120 mg). Intermediate 121A: off-white solid; 99.6% chiral purity; MS (LCMS) 704.6 [M+H]+. Intermediate 121B: off-white solid; 99.2% chiral purity; MS (LCMS) 704.3 [M+H]+. The absolute stereochemistry of Intermediate 121A and Intermediate 121B was arbitrarily assigned. Example 11A (Ra)-(+)-(Z)-16-Chloro-96-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0228] Example 11
Figure imgf000111_0001
mediate 121A following a similar procedure for the preparation of Example 1. Example 11A: (70 mg, 60%), off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.30 (br s, 1H), 8.27-8.22 (m, 1H), 7.58-7.52 (m, 2H), 7.38-7.32 (m, 2H), 6.83-6.78 (m, 2H), 4.79 (s, 1H), 4.08-3.90 (m, 4H), 3.80 (s, 3H), 3.62- 3.52 (m, 2H), 3.50 (s, 3H), 3.45-3.33 (m, 4H), 3.20-3.10 (m, 3H), 2.45-2.35 (m, 1H), 2.25-2.10 (m, 1H), 1.85 (s, 3H); MS (LCMS) 690.7 [M+H]+. The absolute stereochemistry of Example 11A was arbitrarily assigned. Example 11B (Sa)-(-)-(Z)-16-Chloro-96-fluoro-11,21,23,61-tetramethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(5,3)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0229] Example 11
Figure imgf000111_0002
mediate 121B following a similar procedure for the preparation of Example 1. Example 11B: (70 mg, 60%), off-white solid; 97.4% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.30 (br s, 1H), 8.27-8.22 (m, 1H), 7.58-7.52 (m, 2H), 7.38-7.32 (m, 2H), 6.84-6.78 (m, 2H), 4.79 (s, 1H), 4.08-3.90 (m, 4H), 3.80 (s, 3H), 3.62- 3.52 (m, 2H), 3.50 (s, 3H), 3.45-3.33 (m, 4H), 3.20-3.10 (m, 3H), 2.45-2.35 (m, 1H), 2.25-2.10 (m, 1H), 1.86 (s, 3H); MS (LCMS) 690.7 [M+H]+. The absolute stereochemistry of Example 11B was arbitrarily assigned. Intermediate 122 Ethyl 5-((acetylthio)methyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate [0230] To a stirred solution
Figure imgf000112_0001
te 47 (15.5 g, 50.2 mmol) in CH3CN (155 mL) was added KSAc (16.66 g, 100.4 mmol) and KI (5.72 g, 50.2 mmol) at 0 °C. The reaction was stirred at rt for 16 h. The reaction was quenched with water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated. The crude residue was purified by flash chromatography (SiO2, 10% EtOAc/Pet. ether) to afford Intermediate 122 (14 g) as an oil. MS (LCMS) 349.3 [M+H]+. Intermediate 123 Ethyl 5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-(4-methoxybenzyl)-1H-pyrazole- 3-carboxylate [0231] To a degassed
Figure imgf000112_0002
22 (8.20 g, 23.6 mmol) in 1,4-dioxane (164 mL):water (41 mL) was added Intermediate 80 (10.98 g, 25.91 mmol), K2CO3 (3.41 g, 24.7 mmol), XPhos (1.01 g, 2.12 mmol) and Pd2(dba)3 (1.08 g, 1.18 mmol). The mixture was degassed with Ar for 15 min. The reaction was heated at 100°C for 3 h. The reaction was cooled to rt, diluted with EtOAc (250 mL) and passed through a Celite pad. The pad was washed with EtOAc (100 mL). The filtrate was washed with brine (200 mL), dried (Na2SO4), filtered and evaporated. The crude residue was purified by flash chromatography (SiO2, 30% EtOAc/Pet. ether) to afford a mixture of Intermediate 123 (8.7 g) and ethyl 5-(((4-acetoxy-7-fluoronaphthalen-2- yl)thio)methyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate as an oil. MS (LCMS) 467.4 [M+H]+ and 509.5 [M+H]+. Intermediate 124 6-Fluoro-3-(((3-(hydroxymethyl)-1-(4-methoxybenzyl)-1H-pyrazol-5-yl)methyl)thio)naphthalen- 1-ol
Figure imgf000112_0003
[0232] To a stirred solution of Intermediate 23 (8.70 g, 18.7 mmol) in THF (300 mL) was added 2.4 M LiAlH4 in THF (11.66 mL, 27.99 mmol) dropwise at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 3 h. Upon completion by TLC, the reaction was cooled to 0 °C and quenched with sat. Na2SO4 (25 mL). The precipitated solid was removed by filtration and the filtrate was dried (Na2SO4), filtered and evaporated. The crude residue was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 124 (4 g, 40%, 2-steps) as a solid. MS (LCMS) 425.4 [M+H]+. Intermediate 125 3-(((3-((Acetylthio)methyl)-1-(4-methoxybenzyl)-1H-pyrazol-5-yl)methyl)thio)-6- fluoronaphthalen-1-yl acetate [0233] Intermediate
Figure imgf000113_0001
hree steps from Intermediate 124 following a similar procedure for the preparation of Intermediate 52. MS (LCMS) 525.4 [M+H]+. Intermediate 126 Methyl 6-chloro-7-(5-((((5-(((7-fluoro-4-hydroxynaphthalen-2-yl)thio)methyl)-1-(4- methoxybenzyl)-1H-pyrazol-3-yl)methyl)thio)methyl)-1,3-dimethyl-1H-pyrazol-4-yl)-3-(3- hydroxypropyl)-1-methyl-1H-indole-2-carboxylate [0234] Intermed
Figure imgf000113_0002
rmediate 119 and Intermediate 125 following a similar procedure from the preparation of Intermediate 24. MS (LCMS) 827.9 [M+H]+. Intermediate 127 Methyl (Z)-16-chloro-96-fluoro-61-(4-methoxybenzyl)-11,21,23-trimethyl-11H,21H,61H-10-oxa- 4,8-dithia-1(7,3)-indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12- carboxylate [0235] Intermedia
Figure imgf000114_0001
termediate 126 following a similar procedure from the preparation of Intermediates 25A and 25B. MS (LCMS) 810.7 [M+H]+. Intermediate 128 Methyl (Z)-16-chloro-96-fluoro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylate [0236] To a stirred
Figure imgf000114_0002
(500 mg, 0.618 mmol) in TFA (5 mL) was added anisole (2.5 mL, 5 vol) at rt. The mixture was stirred at 100 °C for 48 h in a sealed tube. The mixture was concentrated and water (20 mL) was added. The precipitated solid was filtered, washed with water (20 mL) and dried. The crude material was triturated with diethyl ether (50 mL) to afford Intermediate 128 (320 mg, 75%) as an off-white solid. MS (ESI) 690.1 [M+H]+. Example 12A (Ra)-(+)-(Z)-16-Chloro-96-fluoro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)- indola-2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid
Figure imgf000114_0003
Example 12B (Sa)-(-)-(Z)-16-Chloro-96-fluoro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola- 2(4,5),6(3,5)-dipyrazola-9(3,1)-naphthalenacyclotridecaphane-12-carboxylic acid [0237] To a stirred
Figure imgf000115_0001
128 (320 mg, 0.460 mmol) in MeOH/THF/H2O (1:1:1, 16 mL) was added LiOH•H2O (292 mg, 6.96 mmol). The reaction was stirred at 70°C for 1 h. Upon completion by TLC, the solvent was evaporated, and the aqueous layer was acidified to pH 6 using 2N aqueous HCl (6 mL). The resulting precipitate was collected by filtration, washed with water (5 mL) and dried under vacuum to afford racemic (Z)-16-chloro- 96-fluoro-11,21,23-trimethyl-11H,21H,61H-10-oxa-4,8-dithia-1(7,3)-indola-2(4,5),6(3,5)- dipyrazola-9(3,1)-naphthalenacyclo-tridecaphane-12-carboxylic acid (290 mg, 93%) as an off- white solid. MS (ESI) 676.4. The atropisomers were separated by chiral SFC chromatography (Chiralpak AS-H (30 x 250 mm), 40% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Example 12A, 98 mg) and peak 2 (Example 12B, 81 mg). Example 12A: off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 12.8 (br s, 2H), 8.22-8.18 (m, 1H), 7.66 (d, J=8.4 Hz, 1H), 7.53 (dd, J=10.0, 8.0 Hz, 1H), 7.40-7.22 (m, 2H), 6.97 (d, J=8.4 Hz, 1H), 6.63 (s, 1H), 4.73 (s, 1H), 4.15-3.95 (m, 3H), 3.86-3.78 (m, 4H), 3.56-3.47 (m, 6H), 3.38-3.32 (m, 1H), 3.19-3.02 (m, 2H), 2.67-2.50 (m, 1H), 2.37-2.35 (m, 1H), 1.87 (s, 3H); MS (LCMS) 676.6 [M+H]+. Example 12B: off-white solid; 98.1% chiral purity; δ 12.8 (br s, 2H), 8.21-8.17 (m, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.53 (dd, J=10.4, 8.0 Hz, 1H), 7.35-7.31 (m, 2H), 6.96 (d, J=8.4 Hz, 1H), 6.62 (s, 1H), 4.76 (s, 1H), 4.15-3.95 (m, 3H), 3.86-3.85 (m, 4H), 3.65-3.42 (m, 6H), 3.28-3.26 (m, 1H), 3.10-3.02 (m, 2H), 2.35-2.32 (m, 1H), 2.30-2.21 (m, 1H), 1.87 (s, 3H) MS (LCMS) 676.6 [M+H]+. The absolute stereochemistry of Example 12A and Example 12B was arbitrarily assigned. Intermediate 129 Ethyl 5-(chloromethyl)-1-ethyl-1H-pyrazole-3-carboxylate [0238] To a stirred solut
Figure imgf000116_0001
l 5-(chloromethyl)-1-ethyl-1H-pyrazole-3- carboxylate (5.00 g, 25.3 mmol) in DCM (50 mL) was added SOCl2 (2.20 mL, 30.3 mmol) at 0 °C. The reaction was stirred at rt for 1 h. Upon completion by TLC, the reaction was diluted with DCM (50 mL) and washed with sat. NaHCO3 (50 mL). The organic layer was washed with brine (50 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 129 (5 g, 92%) as an oil. (LCMS) 217.1 [M+H]+. Intermediate 130 Methyl 1-ethyl-5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1H-pyrazole-3-carboxylate [0239] To a stirred sol
Figure imgf000116_0002
29 (5.00 g, 23.1 mmol) in MeOH (50 mL) was added K2CO3 (7.66 g, 55.6 mmol) at rt. The reaction was degassed with Ar for 10 min. In a separate flask, Intermediate 11 (6.01 g, 23.1 mmol) in methanol (10 mL) was degassed with Ar for 10 min. The solution of Intermediate 11 was added to the mixture of Intermediate 129 and K2CO3 in MeOH. The reaction was stirred at rt for 1h. Upon completion, the solvent was evaporated, and the reaction was diluted with water (50 mL). The mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were separated, dried (Na2SO4), filtered and evaporated. The crude material was triturated with DCM (50 mL). The solid was filtered and dried under high vacuum to afford Intermediate 130 (6 g, 76%) as an off-white solid. MS (LCMS) 343.3 [M+H]+. Intermediate 131 3-(((3-((Acetylthio)methyl)-1-ethyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-yl acetate
Figure imgf000116_0003
[0240] Intermediate 131 was prepared in four steps from Intermediate 130 following a similar procedure for the preparation of Intermediate 115. Intermediate 132 Methyl 4-bromo-5-chloro-3-methyl-1H-indole-2-carboxylate [0241] A solution of Na
Figure imgf000117_0001
mmol) in water (100 mL) was added dropwise to a stirred solution of 3-bromo-4-chloroaniline (100 g, 484 mmol) in 35% conc. HCl (500 mL) at 0 °C. The reaction was stirred at 0 °C for 30 min. SnCl2 (275.4 g, 1453 mmol) in water (250 mL) was added at 0 °C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the reaction was filtered. The solid was suspended in sat. NaHCO3 (2 L). The mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (1 L), dried (Na2SO4) and concentrated to afford (3-bromo-4- chlorophenyl)hydrazine (80.5 g 75%) as a yellow solid. MS (LCMS) 223.2 [M+H]+ [0242] To a stirred solution of (3-bromo-4-chlorophenyl)hydrazine (80.0 g, 364 mmol) in MeOH (800 mL) was added conc. H2SO4 (80 mL) followed by dropwise addition of methyl 2- oxobutanoate (30.7 g, 364 mmol) at 0 °C. The reaction was stirred at 60 °C for 24 h. Upon completion by TLC, the MeOH was evaporated. Water (300 mL) was added, and the precipitated solid was filtered. The filter cake was washed with water (500 mL) and dried to afford Intermediate 132 (70 g) as yellow solid. MS (LCMS) 304.2 [M+H]+. Intermediate 133 Methyl 4-bromo-5-chloro-1-(3-ethoxy-3-oxopropyl)-3-methyl-1H-indole-2-carboxylate [0243] To a stirred soluti
Figure imgf000117_0002
32 (70.0 g, 232 mmol) in ACN (700 mL) was added K2CO3 (63.9 g, 464 mmol) at rt. The mixture was stirred for 30 min. Ethyl 3- bromopropanoate (41.9 g, 232 mmol) was added, and the reaction was stirred at 80 °C for 2 d. The suspension was filtered, and the filtrate was evaporated under reduced pressure. The crude residue was dissolved in 10% MeOH:DCM (1000 mL) and washed with water (500 mL). The organic layer was dried (Na2SO4) and evaporated to afford Intermediate 133 (110 g crude) as a brown solid. MS (LCMS) 404.1 [M+H]+. Intermediate 134 Methyl 1-(3-acetoxypropyl)-4-bromo-5-chloro-3-methyl-1H-indole-2-carboxylate [0244] To a stirred soluti
Figure imgf000118_0001
133 (110 g, 274 mmol,) in THF (1.1 L) was added 1M BH3 in THF (1.37 L, 1.37 mol) at 0°C. The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, the solvent was evaporated. Water (1 L) was added to the residue with stirring. The mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (1 L), dried (Na2SO4), filtered and concentrated. The crude material was purified by flash chromatography (SiO2, 15% EtOAc/Pet. ether) to afford methyl 4-bromo-5-chloro-1-(3-hydroxypropyl)-3-methyl-1H-indole-2-carboxylate (14 g, 11%, 3- steps) as an off-white solid. MS (LCMS) 362.1 [M+H]+. [0245] To a stirred solution of methyl 4-bromo-5-chloro-1-(3-hydroxypropyl)-3- methyl-1H-indole-2-carboxylate (7.00 g, 19.4 mmol) in DCM (70 mL) at 0 °C was added Et3N (4.00 mL, 29.1 mmol), Ac2O (2.37 g, 23.3 mmol) and DMAP (1.20 g, 9.71 mmol). The ice bath was removed, and the reaction was stirred at rt for 16 h. Upon completion by TLC, water (200 mL) was added, and the mixture was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (400 mL), dried (Na2SO4) and concentrated under reduced pressure. Two equivalent reactions were run and combined for purification. The crude material was purified by flash chromatography (SiO2, 15% EtOAc/Pet. ether) to afford Intermediate 134 (14 g, 89%) as a white solid. MS (LCMS) 404.3 [M+H]+.
Intermediate 135
Methyl l-(3-acetoxypropyl)-5-chloro-4-(2-(((4-methoxybenzyl)oxy)methyl)-5,6-dihydro-4H- pyrrolo[l,2-b]pyrazol-3-yl)-3-methyl-lH-indole-2-carboxylate
Figure imgf000119_0001
[0246] Intermediate 135 was synthesized from Intermediate 134 and Intermediate
96 following a similar procedure from the preparation of Intermediate 20. MS (LCMS) 580.7 [M+H]+.
Intermediate 136
Methyl l-(3-acetoxypropyl)-5-chloro-4-(2-(hydroxymethyl)-5,6-dihydro-4H-pyrrolo[l,2- b]pyrazol-3-yl)-3-methyl-lH-indole-2-carboxylate
Figure imgf000119_0002
[0247] Intermediate 136 was synthesized from Intermediate 135 following a similar procedure for the preparation of Intermediate 21. MS (LCMS) 460.5 [M+H]+.
Intermediate 137
Methyl l-(3-acetoxypropyl)-5-chloro-4-(2-(chloromethyl)-5,6-dihydro-4H-pyrrolo[l,2- b]pyrazol-3-yl)-3-methyl-lH-indole-2-carboxylate
Figure imgf000119_0003
[0248] Intermediate 137 was synthesized from Intermediate 136 following a similar procedure for the preparation of Intermediate 22. MS (LCMS) 478.3 [M+H]+. Intermediate 138 Methyl 1-(3-acetoxypropyl)-5-chloro-4-(2-(iodomethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol- 3-yl)-3-methyl-1H-indole-2-carboxylate [0249] Intermediate 138
Figure imgf000120_0001
m Intermediate 137 following a similar procedure for the preparation of Intermediate 23. MS (LCMS) 570.4 [M+H]+. Intermediate 139 Methyl 5-chloro-4-(2-((((1-ethyl-5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1H-pyrazol-3- yl)methyl)thio)methyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)-1-(3-hydroxypropyl)-3- methyl-1H-indole-2-carboxylate [0250] Intermed
Figure imgf000120_0002
rmediate 138 and Intermediate 131 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 730.3 [M+H]+. Intermediate 140A (Ra)-(+)-Methyl (Z)-15-chloro-61-ethyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000120_0003
Intermediate 140B (Sa)-(-)-Methyl (Z)-15-chloro-61-ethyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate [0251] Intermediates
Figure imgf000121_0001
ynthesized from Intermediate 139 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-15-chloro-61-ethyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia-1(4,1)- indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)-naphthalenacyclotridecaphane-12- carboxylate (160 mg, 36%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OJ-3, (30 x 250 mm), 30% MeOH) to give peak 1 (Intermediate 140A, 56 mg) and peak 2 (Intermediate 140B, 56 mg). Intermediate 140A: off-white solid; 99.7% chiral purity; MS (LCMS) 712.6 [M+H]+. Intermediate 140B: off-white solid; 99.6% chiral purity; MS (LCMS) 712.6 [M+H]+. The absolute stereochemistry of Intermediate 140A and Intermediate 140B was arbitrarily assigned. Example 13A (Ra)-(+)-(Z)-15-Chloro-61-ethyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia-1(4,1)- indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)-naphthalenacyclotridecaphane-12- carboxylic acid [0252] Example 13A
Figure imgf000121_0002
rmediate 140A following a similar procedure for the preparation of Example 1. Example 13A: (50 mg, 92%), off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.33 (br s, 1H), 8.13 (d, J=7.6 Hz, 1H), 7.75- 7.65 (m, 2H), 7.53-7.42 (m, 2H), 7.36 (s, 1H), 7.19 (d, J=8.8 Hz, 1H), 6.67 (s, 1H), 5.08-5.00 (m, 1H), 4.84 (s, 1H), 4.62-4.52 (m, 1H), 4.20 (q, J=15.6 Hz, 2H), 4.10-3.95 (m, 5H), 3.90-3.80 (m, 1H), 3.35-3.25 (m, 1H), 3.23-3.10 (m, 2H), 2.96 (d, J=14.4 Hz, 1H), 2.60-2.50 (m, 4H), 2.40-2.30 (m, 2H), 2.04 (s, 3H), 1.29 (t, J=7.0 Hz, 3H); MS (LCMS) 698.6 [M+H]+. The absolute stereochemistry of Example 13A was arbitrarily assigned. Example 13B (Sa)-(-)-(Z)-15-Chloro-61-ethyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia-1(4,1)- indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)-naphthalenacyclotridecaphane-12- carboxylic acid [0253] Example 13B
Figure imgf000122_0001
rmediate 140B following a similar procedure for the preparation of Example 1. Example 13B: (49 mg, 90%), off-white solid; 99.4% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.35 (br s, 1H), 8.13 (d, J=7.6 Hz, 1H), 7.75- 7.65 (m, 2H), 7.53-7.42 (m, 2H), 7.36 (s, 1H), 7.19 (d, J=8.8 Hz, 1H), 6.67 (s, 1H), 5.08-5.00 (m, 1H), 4.83 (s, 1H), 4.62-4.52 (m, 1H), 4.20 (q, J=15.6 Hz, 2H), 4.10-3.95 (m, 5H), 3.90-3.80 (m, 1H), 3.35-3.25 (m, 1H), 3.23-3.10 (m, 2H), 2.96 (d, J=14.4Hz, 1H), 2.60-2.50 (m, 4H), 2.40-2.30 (m, 2H), 2.04 (s, 3H), 1.29 (t, J=7.0 Hz, 3H); MS (LCMS) 698.6 [M+H]+. The absolute stereochemistry of Example 13B was arbitrarily assigned. Intermediate 141 Diethyl 1-isopropyl-1H-pyrazole-3,5-dicarboxylate [0254] To an ice cooled, st
Figure imgf000122_0002
of diethyl 1H-pyrazole-3,5-dicarboxylate (50.0 g, 236 mmol) in DMF (250 mL) was added 60% NaH (10.3 g, 259 mmol) portionwise. The mixture was stirred at 0 oC for 30 min. Iodopropane (44.0 g, 259 mmol) was added slowly. The reaction was stirred at rt for 2 h. Upon completion by TLC, the reaction was quenched with water (500 mL). The solid was collected by filtration and washed with water (2 x 100 mL). The solid was dried under high vacuum to afford Intermediate 141 (40 g) as an off-white solid. MS (LCMS) 255.2 [M+H]+. Intermediate 142 3-(Ethoxycarbonyl)-1-isopropyl-1H-pyrazole-5-carboxylic acid [0255] To a stirred solution
Figure imgf000123_0001
ate 141 (40.0 g, 157 mmol) in EtOH (100 mL) was added 2M KOH in ethanol (120 mL, 240 mmol). The reaction was stirred at rt for 48 h. Upon completion by TLC, the reaction was diluted with 2N HCl until the pH was 2. The mixture was extracted with EtOAc (2 x 400 mL). The combined organic layers were washed with brine (600 mL), dried (Na2SO4), filtered and evaporated to afford Intermediate 142 (30 g) as an off- white solid. MS (LCMS) 227.2 [M+H]+. Intermediate 143 Ethyl 5-(hydroxymethyl)-1-isopropyl-1H-pyrazole-3-carboxylate [0256] To a stirred solution
Figure imgf000123_0002
e 142 (10.0 g, 44.2 mmol) in THF (100 mL) at -10 oC was added DIPEA (11.42 g, 88.40 mmol) and isobutyl chloroformate (12.0 mL, 88.4 mmol). The ice bath was removed, and the reaction was stirred at rt for 1 h. Upon completion by LCMS, the reaction was cooled to -10oC and NaBH4 (3.90 g, 106 mmol) was added. The reaction was stirred at rt for 16 h. Upon completion, the pH was adjusted to 2 with 2N HCl. The mixture was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (400 mL), dried (Na2SO4), filtered and evaporated. The crude material was purified by flash chromatography (SiO2, 50% EtOAc/Pet. ether) to afford Intermediate 143 (2 g, 12%, 3-steps) as a yellow solid. MS (LCMS) 213.1 [M+H]+. Intermediate 144 Ethyl 5-(chloromethyl)-1-isopropyl-1H-pyrazole-3-carboxylate
Figure imgf000123_0003
[0257] To a stirred solution of Intermediate 143 (7.00 g, 33.0 mmol) in DCM (90 mL) under Ar was added SOCl2 (2.60 mL, 36.3 mmol) at 0 °C, and the reaction was stirred at rt for 1 h. The reaction was diluted with DCM (250 mL) and washed with sat. NaHCO3 (3 x 200 mL). The reaction was repeated on an equal scale and combined for the work up. The organic layer was dried (Na2SO4), filtered and evaporated to afford Intermediate 144 (14.5 g) as an off-white solid. MS (LCMS) 231.1 [M+H]+. Intermediate 145 Methyl 5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1-isopropyl-1H-pyrazole-3-carboxylate [0258] To a stirred so
Figure imgf000124_0001
4 (7.00 g, 30.3 mmol) in MeOH (60 mL) was added K2CO3 (10.0 g, 72.8 mmol) and the mixture was degassed with A for 10 min. In a separate flask, Intermediate 11 (8.68 g, 33.9 mmol) in MeOH (15 mL) was degassed with Ar for 10 min. The solution of Intermediate 11 in MeOH was added dropwise to the mixture of Intermediate 144 and K2CO3. The reaction was stirred at rt for 16 h. The solvent was evaporated, and the reaction was diluted with water (200 mL). The mixture was extracted with EtOAc (2 x 200 mL). The combined organic layers were separated, dried (Na2SO4), filtered and evaporated. Two reactions of equal size were run simultaneously and combined for purification. The crude material was purified by flash chromatography (SiO2, 20% EtOAc/Pet. ether) to afford Intermediate 145 (18 g, 77%, 2-steps) as a brown solid. MS (LCMS) 357.1 [M+H]+. Intermediate 146 3-(((3-((Acetylthio)methyl)-1-isopropyl-1H-pyrazol-5-yl)methyl)thio)naphthalen-1-yl acetate [0259] Intermediate 1
Figure imgf000124_0002
eps from Intermediate 145 following a similar procedure for the preparation of Intermediate 115. MS (LCMS) 429.1 [M+H]+. Intermediate 147 Methyl 5-chloro-4-(2-((((5-(((4-hydroxynaphthalen-2-yl)thio)methyl)-1-isopropyl-1H-pyrazol-3- yl)methyl)thio)methyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)-1-(3-hydroxypropyl)-3- methyl-1H-indole-2-carboxylate OH
Figure imgf000125_0001
146 following a similar procedure for the preparation of Intermediate 24. MS (LCMS) 744.4 [M+H]+. Intermediate 148A (Ra)-(+)-Methyl (Z)-15-chloro-61-isopropyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8- dithia-1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (Sa)-(-)-M
Figure imgf000125_0002
dithia-1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate
Figure imgf000125_0003
[0261] Intermediates 148A and 148B were synthesized from Intermediate 147 following a similar procedure for the preparation of Intermediates 25A and 25B to afford racemic methyl (Z)-15-chloro-61-isopropyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo-[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylate (75 mg, 10%). The atropisomers were separated by chiral SFC chromatography (Chiralcel-OJ-3 (30 x 250 mm), 20% (0.2% NH3 in MeOH in CH3CN:MeOH; 1:1)) to give peak 1 (Intermediate 148A, 25 mg) and peak 2 (Intermediate 148B, 25 mg). Intermediate 148A: off-white solid; 99.9% chiral purity; MS (LCMS) 726.2 [M+H]+. Intermediate 148B: off-white solid; 99.2% chiral purity; MS (LCMS) 726.2 [M+H]+. The absolute stereochemistry of Intermediate 148A and Intermediate 148B was arbitrarily assigned. Example 14A (Ra)-(+)-(Z)-15-Chloro-61-isopropyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0262] Example 14A
Figure imgf000126_0001
rmediate 148A following a similar procedure for the preparation of Example 1. Example 14A: (20 mg, 83%), off-white solid; 99.9% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.4 (br s, 1H), 8.11 (d, J=7.6 Hz, 1H), 7.74-6.70 (m, 2H), 7.50-7.44 (m, 2H), 7.33 (s, 1H), 7.19 (d, J=8.8 Hz, 1H), 6.69 (s, 1H), 5.09-5.01 (m, 1H), 4.82 (s, 1H), 4.62-4.56 (m, 2H), 4.32-4.18 (m, 2H), 4.10-4.08 (m, 2H), 4.00-3.96 (m, 1H), 3.88- 3.86 (m, 1H), 3.17-3.13 (m, 3H), 2.97 (d, J=14.0 Hz, 1H), 2.54-2.50 (m, 5H), 2.37-2.32 (m, 1H), 2.03 (s, 3H), 1.37-1.28 (m, 6H); MS (LCMS) 712.8 [M+H]+. The absolute stereochemistry of Example 14A was arbitrarily assigned. Example 14B (Sa)-(-)-(Z)-15-Chloro-61-isopropyl-13-methyl-25,26-dihydro-11H,24H,61H-10-oxa-4,8-dithia- 1(4,1)-indola-2(3,2)-pyrrolo[1,2-b]pyrazola-6(3,5)-pyrazola-9(3,1)- naphthalenacyclotridecaphane-12-carboxylic acid [0263] Example 14B
Figure imgf000127_0001
rmediate 148B following a similar procedure for the preparation of Example 1. Example 14B: (20 mg, 83%), off-white solid; 99.2% chiral purity; 1H NMR (400 MHz, DMSO-d6) δ 13.4 (br s, 1H), 8.11 (d, J=7.6 Hz, 1H), 7.72-7.69 (m, 2H), 7.50-7.44 (m, 2H), 7.33 (s, 1H), 7.19 (d, J=9.2 Hz, 1H), 6.68 (s, 1H), 5.09-5.01 (m, 1H), 4.80 (s, 1H), 4.60-4.55 (m, 2H), 4.31-4.18 (m, 2H), 4.10-4.08 (m, 2H), 4.00-3.96 (m, 1H), 3.87- 3.86 (m, 1H), 3.17-3.13 (m, 3H), 2.97 (d, J=14.0 Hz, 1H), 2.53-2.50 (m, 5H), 2.37-2.32 (m, 1H), 2.03 (s, 3H), 1.34 (d, J=8.0 Hz, 3H), 1.29 (d, J=6.4 Hz, 3H); MS (LCMS) 712.6 [M+H]+. The absolute stereochemistry of Example 14B was arbitrarily assigned. Example A Mcl-1 Homogeneous Time Resolved Fluorescence (HTRF) Assay [0264] Binding to Bcl-2 proteins Mcl-1 was assessed using an HTRF assay. Background: FAM-Bak/Bad binds to surface pocket of the Bcl-2 protein family. This binding can be monitored by HTRF signals between anti-GST-Tb and FAM-peptide using GST-tagged Bcl proteins. Assay conditions: 4 nM Mcl-1, 100 nM FAM-Bak peptide, in 20 mM K Phosphate, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.005% Triton X-100 and 1% DMSO (final). Assay procedure: Compounds were tested in 10-dose IC50 mode, in singlicate, with 3-fold serial dilution starting at 10µM or 1 µM. Compound stock solutions were added to protein solution using Acoustic technology. The compounds were then incubated with protein for 10 min at rt. The respective FAM labeled peptide was added and incubated for another 10 min. Anti-GST-Tb was added. After 60 min at rt, the HTRF fluorescence signal ratio was measured. Curve fits were performed in GraphPad Prism 4 with “sigmoidal dose-response (variable slope)”; 4 parameters with Hill Slope. The results are shown in Table 1. Example B NCI-H929 Cell Proliferation Assay [0265] Cell proliferation was measured using the CellTiter-Glo® Luminescent Cell Viability Assay. The assay involved the addition of a single reagent (CellTiter-Glo® Reagent) directly to cells cultured in serum-supplemented medium. NCI-H929 (ATCC CRL-9068) cells were cultured according to ATCC recommendations and were seeded at 3,000 cells per well. [0266] Each compound evaluated was prepared as a DMSO stock solution (10 mM). Compounds were tested in duplicate on each plate, with a 10-point serial dilution curve (1:3 dilution). Compound treatment (1.0 μL) was added from the compound dilution plate to the cell plate. The highest compound concentration was 10 μM (final), with a 0.1% final DMSO concentration. Plates were then incubated at 37 °C, 5% CO2. After 72 h of compound treatment, cell plates were equilibrated at rt for approximately 30 mins. An equi-volume amount of CellTiter- Glo® Reagent (40 μL) was added to each well. Plates were mixed for 2 mins on an orbital shaker to induce cell lysis and then incubated at rt for 10 mins to stabilize the luminescent signal. Luminescence was recorded using an Envision plate reader according to CellTiter-Glo protocol. IC50 of each compound was calculated using GraphPad Prism by nonlinear regression analysis. IC50 values are provided in Table 1. Table 1 Examples Mcl 1 IC50 (nM) H929 IC50 (nM)
Figure imgf000128_0001
Examples Mcl-1 IC50 (nM) H929 IC50 (nM) gle
Figure imgf000129_0001
. For H929 CTG IC50: A = a single IC50 ≤ 100 nM; B = a single IC50 >100 nM and < 1000 nM; C = a single IC50 ≥1000 nM. [0267] Furthermore, although the foregoing has been described in some detail by way of illustrations and examples for purposes of clarity and understanding, it will be understood by those of skill in the art that numerous and various modifications can be made without departing from the spirit of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but rather to also cover all modification and alternatives coming with the true scope and spirit of the present disclosure.

Claims

WHAT IS CLAIMED IS: 1. A compound of Formula (I), or a pharmaceutically salt thereof, wherein the compound has the structure: wherein:
Figure imgf000130_0001
R1 is fluoro, chloro or methyl; R2 is hydrogen or fluoro;
Figure imgf000130_0002
unsubstituted C1-4 alkyl; , wherein R6 and R7 are independently hydrogen or
Figure imgf000130_0003
provided that when R1 is Cl; and Ring A
Figure imgf000130_0004
provide that when R1 is Cl; Rin ; Ring B , then R2 cannot be fluoro.
Figure imgf000131_0001
2. The compound of Claim
Figure imgf000131_0005
, uoro. 3. The compound of Claim 1, wherein R1 is chloro. 4. The compound of Claim 1, wherein R1 is methyl. 5. The compound of any one of Claims 1-4, wherein R2 is fluoro. 6. The compound of any one of Claims 1-4, wherein R2 is hydrogen. 7. The compound of any one of Claims 1-6, wherein Ring B . 8. The compound of Claim 7, wherein R6 is hydrogen.
Figure imgf000131_0002
9. The compound of Claim 7, wherein R6 is methyl. 10. The compound of Claim 7, wherein R6 is ethyl. 11. The compound of Claim 7, wherein R6 is n-propyl or iso-propyl. 12. The compound of Claim 7, wherein R6 is n-butyl, sec-butyl or tert-butyl. 13. The compound of any one of Claims 1-6, wherein Rin . 14. The compound of Claim 13, wherein R7 is hydrogen.
Figure imgf000131_0003
15. The compound of Claim 13, wherein R7 is methyl. 16. The compound of Claim 13, wherein R7 is ethyl. 17. The compound of Claim 13, wherein R7 is n-propyl or iso-propyl. 18. The compound of Claim 13, wherein R7 is n-butyl, sec-butyl or tert-butyl. 19. The compound of any one of Claims 1-18, wherein Rin .
Figure imgf000131_0004
20. The compound of Claim 19, wherein Rin .
Figure imgf000132_0001
21. The compound of Claim 19, wherein Rin . 22. The compound of any one of Claims 19-2
Figure imgf000132_0002
thyl. 23. The compound of any one of Claims 19-21, wherein R3 is ethyl. 24. The compound of any one of Claims 19-21, wherein R3 is n-propyl or iso-propyl. 25. The compound of any one of Claims 19-21, wherein R3 is n-butyl, sec-butyl or tert- butyl. 26. The compound of any one of Claims 19-25, wherein R4 is methyl. 27. The compound of any one of Claims 19-25, wherein R4 is ethyl. 28. The compound of any one of Claims 19-25, wherein R4 is n-propyl or iso-propyl. 29. The compound of any one of Claims 19-25, wherein R4 is n-butyl, sec-butyl or tert- butyl. 30. The compound of any one of Claims 1-18, wherein Rin .
Figure imgf000132_0003
31. The compound of Claim 30, wherein Rin .
Figure imgf000132_0004
32. The compound of Claim 30, wherein Rin .
Figure imgf000132_0005
33. The compound of any one of Claims 1-18, wherein Rin .
Figure imgf000132_0006
43. The compound of Claim 1, wherein the compound is selected from the group consisting of:
Figure imgf000134_0001
Figure imgf000135_0001
Figure imgf000136_0002
Figure imgf000136_0001

Figure imgf000137_0001
Figure imgf000138_0001
Figure imgf000139_0001
44. The compound of Claim 1, wherein the compound is selected from the group consisting of:
Figure imgf000140_0001
Figure imgf000141_0001
Figure imgf000142_0001
Figure imgf000143_0001
Figure imgf000144_0001
Figure imgf000145_0001
Figure imgf000146_0001

Figure imgf000147_0001
Figure imgf000148_0001

Figure imgf000149_0001
Figure imgf000150_0001
Figure imgf000151_0001
pharmaceutically acceptable salt of any of the foregoing.
45. A compound is selected from the group consisting of:
Figure imgf000152_0001
pharmaceutically acceptable salt of any of the foregoing.
46. A pharmaceutical composition comprising an effective amount of the compound of any one of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.
47. Use of an effective amount of a compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 in the manufacture of a medicament for ameliorating or treating a cancer, wherein the cancer is selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a nonHodgkin’s lymphoma.
48. Use of an effective amount of a compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 in the manufacture of a medicament for inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma.
49. Use of an effective amount of a compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 in the manufacture of a medicament for ameliorating or treating a malignant growth or tumor, wherein the malignant growth or tumor is due to a cancer selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma.
50. Use of an effective amount of a compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 in the manufacture of a medicament for inhibiting the activity of Mcl- 1.
51. A compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 for use in ameliorating or treating a cancer, wherein the cancer is selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a non-Hodgkin’s lymphoma.
52. A compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 for use in inhibiting replication of a malignant growth or a tumor, wherein the malignant growth or tumor is due to a cancer selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer, a non-small cell cancer, a breast cancer, a lung cancer , a stomach cancer, a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a nonHodgkin’s lymphoma.
53. A compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 for use in ameliorating or treating a malignant growth or tumor, wherein the malignant growth or tumor is due to a cancer selected from a brain cancer, a cervicocerebral cancer, an esophageal cancer, a thyroid cancer, a small cell cancer a non-small cell cancer a breast cancer a lung cancer a stomach cancer a gallbladder/bile duct cancer, a liver cancer, a pancreatic cancer, a colon cancer, a rectal cancer, an ovarian cancer, a choriocarcinoma, an uterus body cancer, an uterocervical cancer, a renal pelvis/ureter cancer, a bladder cancer, a prostate cancer, a penis cancer, a testicular cancer, a fetal cancer, Wilms' cancer, a skin cancer, malignant melanoma, a neuroblastoma, an osteosarcoma, an Ewing's tumor, a soft part sarcoma, an acute leukemia, a chronic lymphatic leukemia, a chronic myelocytic leukemia, polycythemia vera, a malignant lymphoma, multiple myeloma, a Hodgkin's lymphoma, and a nonHodgkin’s lymphoma.
54. A compound of any one of Claims 1-45, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of Claim 46 for use in for inhibiting the activity of Mcl- 1.
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