Conjugates of chemotherapeutic agents and tissue-binding small molecules, compositions, and methods thereof

Abstract

The present invention provides novel conjugates of tissue-binding small molecules and therapeutic agents and pharmaceutical compositions thereof, as well as their use in and methods of treating certain diseases or disorders, such as cancer.

Description

[Technical field]

[0001] The present invention relates generally to novel compounds and their therapeutic uses. More specifically, the present invention provides conjugates of tissue-binding small molecules and therapeutic agents (e.g., anti-cancer agents) and pharmaceutical compositions thereof, as well as their use and methods of treatment in the treatment of certain diseases or disorders (e.g., cancer).

[0002] Immunotherapy, which approaches the treatment of disease by activating or suppressing a patient's immune system, has garnered great interest from researchers and clinicians over the past decade, especially due to its promise in treating various forms of cancer. (Syn,et al.2017 The Lancet Oncol.18(12):e731*e741; Conforti L 2012 Clin.Immunol.142(2):105*106; Nishino,et al.2017 Nat.Rev.Clin.Oncol.14(11):655-668.) To improve treatment outcomes, immunotherapeutic treatments need to be combined with small molecule drugs such as chemotherapeutic agents, kinase inhibitors, indoleamine-2,3-dioxygenase 1 (IDO-1) and adenosine receptor inhibitor (A2a) inhibitors, chemokine receptor antagonists, Toll-like receptor (TLR) and stimulator of interferon genes (STING) modulators. (Huck,et al.2018 Angew Chem.Int.Ed.57,4412-4428.)

[0003] Small molecule chemotherapy drugs are an important part of conventional cancer treatment and may be combined with surgery, radiation therapy, and immunotherapy to improve clinical outcomes. The challenge of conventional chemotherapy is to maintain efficacy while reducing or avoiding side effects and toxicity resulting from systemic exposure. In addition, some drugs must be frequently dosed by intravenous (IV) injections or infusions for hours. Patient compliance and associated hospital costs can also be a challenge.

[0004] Thus, there is an ongoing need for new cancer treatments, particularly those that offer good efficacy and safety profiles. Summary of the Invention

[0005] The present invention is based in part on the unexpected discovery of novel small molecule compounds, methods for their synthesis, and pharmaceutical compositions and methods thereof for treating or ameliorating various diseases or disorders.

[0006] In one aspect, the present invention generally relates to a compound having the following formula:

[0007] [ka] wherein X is CHR or NR, and R is H or alkyl;

[0008] [ka] (wherein n is an integer selected from 2 to 8 (i.e., 2, 3, 4, 5, 6, 7, or 8));

[0009] [ka] (wherein n is an integer selected from 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10));

[0010] [ka] (In the formula, R 2 is H or P(=O)(R 3 )2, and R 3 is alkyl and L is a single bond, or

[0011] [ka] is a group selected from k is an integer selected from 0 to 4 (e.g., 0, 1, 2, 3, or 4);

[0012] [ka] (In the formula, R 2 is H or (=O)(R 3 )2, and R 3 is alkyl and L is a single bond, or

[0013] [ka] is a group selected from k is an integer selected from 0 to 4 (e.g., 0, 1, 2, 3, or 4);

[0014] [ka]

[0015] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0016] In yet another aspect, the invention generally relates to pharmaceutical compositions comprising a compound disclosed herein and a pharma- ceutically acceptable excipient, carrier, or diluent.

[0017] In yet another aspect, the invention generally relates to unit dosage forms that include pharmaceutical compositions that include the compounds disclosed herein.

[0018] In yet another aspect, the invention generally relates to a method of treating or reducing a disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein and a pharma- ceutical acceptable excipient, carrier, or diluent.

[0019] In yet another aspect, the invention generally relates to the use of the compounds disclosed herein for the treatment or amelioration of a disease or disorder, such as cancer.

[0020] In yet another aspect, the invention generally relates to the use of a compound disclosed herein, and a pharma- ceutically acceptable excipient, carrier, or diluent, in the preparation of a medicament for the treatment or amelioration of a disease or disorder, such as cancer.

[0021] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. General principles of organic chemistry, as well as specific functional moieties and reactivities, are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

[0022] Certain compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds as being within the scope of the present invention, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof. Additional asymmetric carbon atoms may be present in a substituent, such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in the present invention.

[0023] Isomeric mixtures containing any of a variety of isomeric ratios may be utilized according to the present invention.For example, when only two isomers are combined, mixtures containing isomeric ratios of 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 are contemplated by the present invention.Those skilled in the art will readily understand that similar ratios are contemplated for more complex isomeric mixtures.

[0024] For example, if a particular enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to yield the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts can be formed with a suitable optically active acid or base, followed by separation of the diastereomers thus formed by fractional crystallization or chromatographic methods well known in the art, followed by recovery of the pure enantiomers.

[0025] Solvates and polymorphs of the compounds of the invention are also contemplated herein. Solvates of the compounds of the invention include, for example, hydrates.

[0026] As used herein, the term "inhibit" refers to any measurable decrease in biological activity. Thus, as used herein, "inhibit" or "inhibition" can refer to a percentage of normal levels of activity.

[0027] As used herein, the term "effective amount" of an active agent refers to the amount sufficient to induce desired biological response.As will be understood by those skilled in the art, the effective amount of the compound of the present invention can vary depending on factors such as desired biological endpoint, pharmacokinetics of the compound, disease to be treated, mode of administration, and patient.

[0028] As used herein, the term "treatment" of a disease or disorder or "treating" thereof refers to a method of reducing, delaying, or ameliorating such a condition before or after such a condition occurs. Treatment may target one or more effects or symptoms of the disease and / or underlying pathology. Treatment may be any reduction, including but not limited to, complete elimination of the disease or symptoms of the disease. Compared to an equivalent untreated control, the degree of such reduction or prevention is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100%, as measured by any standard technique.

[0029] As used herein, a "pharmaceutically acceptable form" of a disclosed compound includes, but is not limited to, pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs, and isotopically labeled derivatives thereof. In one embodiment, a "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, prodrugs, and isotopically labeled derivatives thereof. In some embodiments, a "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable isomers and stereoisomers, prodrugs, and isotopically labeled derivatives thereof.

[0030] In certain embodiments, the pharma- ceutically acceptable form is a pharma- ceutically acceptable salt. As used herein, the term "pharma- ceutically acceptable salt" refers to such salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of a subject without undue toxicity, irritation, allergic reaction, etc., commensurate with a reasonable benefit-risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharma- ceutically acceptable salts in detail in J.Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic and organic acids and bases. Examples of pharma- ceutically acceptable, non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloride, hydrobromide, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharma- ceutically acceptable salts are adipate, alginate, ascorbate, aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, Examples of suitable salts include lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.In some embodiments, organic acids from which salts may be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoroacetic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0031] The salts can be prepared in situ during the isolation and purification of the disclosed compounds, or separately, for example, by reacting the free base or free acid of the parent compound with a suitable base or acid, respectively. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N + (C 1-4 Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations, formed where appropriate using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, base ion exchange resins, and the like, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salts can be selected from ammonium, potassium, sodium, calcium, and magnesium salts.

[0032] In certain embodiments, the pharma- ceutically acceptable form is a "solvate" (e.g., a hydrate). As used herein, the term "solvate" refers to a compound that further comprises a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. The solvate may be of the disclosed compound or a pharma- ceutically acceptable salt thereof. When the solvent is water, the solvate is a "hydrate". Pharmaceutically acceptable solvates and hydrates are complexes that may contain, for example, 1 to about 100, 1 to about 10, 1 to about 2, about 3, or about 4 solvent or water molecules. It will be understood that the term "compound" as used herein encompasses compounds and solvates of compounds, as well as mixtures thereof.

[0033] In certain embodiments, the pharma- ceutically acceptable form is a prodrug. As used herein, the term "prodrug" (or "pro-drug") refers to a compound that is transformed in vivo to yield the disclosed compound or a pharma- ceutically acceptable form of the compound. A prodrug may be inactive when administered to a subject, but is converted in vivo to an active compound, for example, by hydrolysis (hydrolysis in blood). In certain cases, a prodrug has improved physical and / or delivery properties over the parent compound. A prodrug can increase the bioavailability of a compound when administered to a subject (e.g., by allowing enhanced absorption into the blood after oral administration) or enhance delivery to a biological compartment of interest (e.g., the brain or lymphatic system) compared to the parent compound. Exemplary prodrugs include derivatives of the disclosed compounds that have enhanced aqueous solubility or active transport through intestinal membranes compared to the parent compound.

[0034] Prodrug compounds often offer advantages of solubility, tissue compatibility, or delayed release in mammals (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam). A discussion of prodrugs is provided in Higuchi, T., et al., "Pro-drugs as Novel Delivery Systems," ACS Symposium Series, Vol. 14, and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are fully incorporated herein by reference. Exemplary advantages of a prodrug can include, but are not limited to, its physical properties, such as enhanced water solubility for parenteral administration at physiological pH compared to the parent compound, or the ability to enhance absorption from the gastrointestinal tract, or enhanced drug stability for long-term storage.

[0035] Prodrugs generally known in the art include well-known acid derivatives, such as esters prepared by reacting a parent acid with a suitable alcohol, amides prepared by reacting a parent acid compound with an amine, basic groups reacted to form acylated base derivatives, and the like. Of course, other prodrug derivatives can be combined with other features disclosed herein to enhance bioavailability. Thus, one skilled in the art will understand that certain of the compounds of the present disclosure that have free amino, amido, hydroxy, or carboxylic acid groups can be converted into prodrugs. Prodrugs include compounds that have an amino acid residue or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues covalently bonded to a free amino, hydroxy, or carboxylic acid group of a compound of the present disclosure via a peptide bond. Amino acid residues include the 20 naturally occurring amino acids, commonly designated by their three letter symbols, and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline, β-alanine, γ-aminobutyric acid, citrulline homocysteine, homoserine, ornithine, and methionine sulfone. Prodrugs also include compounds having a carbonate, carbamate, amide, or alkyl ester moiety covalently bonded to any of the above substituents disclosed herein.

[0036] As used herein, the term "pharmaceutical acceptable" excipient, carrier, or diluent refers to a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, involved in the carrying or transport of a medicinal agent of interest from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials which can serve as pharma- ceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; celluloses and their derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; excipients such as powdered tragacanth, malt, gelatin, talc, cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar-agar, buffers such as magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffers, and other non-toxic compatible substances utilized in pharmaceutical preparations. Wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate, magnesium stearate, and polyethylene oxide-polypropylene oxide copolymers, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may also be present in the composition.

[0037] As used herein, the terms "isolated" or "purified" refer to material that is substantially or essentially free from components that normally accompany it in its native state. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography.

[0038] As used herein, the term "subject" refers to any animal (e.g., mammal), including but not limited to humans, non-human primates, rodents, etc., that will be the recipient of a particular treatment. Typically, the terms "subject" and "patient" are used interchangeably herein in reference to a human subject.

[0039] As used herein, the term "low dosage" refers to a dosage that is at least 5% less (e.g., at least 10%, 20%, 50%, 80%, 90%, or even 95%) than the lowest standard recommended dosage of a particular compound formulated for a given route of administration for the treatment of any human disease or disorder. For example, a low dosage of a drug formulated for administration by inhalation will be different from a low dosage of the same drug formulated for oral administration.

[0040] As used herein, the term "high dose" means at least 5% (e.g., at least 10%, 20%, 50%, 100%, 200%, or even 300%) more than the highest standard recommended dose of a particular compound for the treatment of any human disease or disorder.

[0041] Isotopically labeled compounds are also within the scope of the present disclosure.As used herein, "isotopically labeled compounds" or "isotopic derivatives" refer to compounds of the present disclosure, including pharmaceutical salts and prodrugs thereof, each of which is described herein, in which one or more atoms are replaced by atoms with atomic masses or mass numbers different from those normally found in nature.Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, for example, each of 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, and 36Examples include Cl.

[0042] By isotopically labelling the compounds of the present disclosure, the compounds may be useful in drug and / or substrate tissue distribution assays. 3 H) and carbon-14 ( 14 C) labeled compounds are particularly preferred due to their ease of preparation and detectability. 2 Substitution with heavier isotopes such as H) can provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in some circumstances. The isotopically labeled compounds of the present disclosure, including pharmaceutical salts, esters, and prodrugs thereof, can be prepared by any means known in the art. The benefits are usually abundant. 12 C 13 C. (See WO2007 / 005643, WO2007 / 005644, WO2007 / 016361 and WO2007 / 016431).

[0043] For example, deuterium ( 2 H) can be incorporated into the compounds disclosed herein for the purpose of manipulating the oxidative metabolism of the compounds through the primary kinetic isotope effect. The primary kinetic isotope effect is the change in the rate of a chemical reaction resulting from the exchange of an isotope nucleus, which in turn is caused by the change in the ground state energy required for covalent bond formation after this isotope exchange. The exchange of a heavier isotope usually results in a lowering of the ground state energy of a chemical bond, thus causing a decrease in the rate of rate-limiting bond breaking. If the bond breaking occurs in or near a saddle point region along the coordinate of a multi-product reaction, it can substantially change the product distribution ratio. For example, if deuterium is attached to a carbon atom in a non-exchangeable position, k M / k DA rate difference of 2-7 is typical. If this rate difference is successfully applied to a compound disclosed herein that is susceptible to oxidation, the in vivo profile of the compound can be dramatically altered, resulting in improved pharmacokinetic properties.

[0044] When discovering and developing therapeutic agents, one skilled in the art can optimize pharmacokinetic parameters while retaining desirable in vitro properties. It is reasonable to assume that many compounds with poor pharmacokinetic profiles are prone to oxidative metabolism. Currently available in vitro liver microsomal assays provide valuable information regarding the course of this type of oxidative metabolism, which in turn allows for the rational design of deuterated compounds of those disclosed herein with improved stability through resistance to such oxidative metabolism. Significant improvements in the pharmacokinetic profiles of the compounds disclosed herein are thereby obtained, including in vivo half-life (t / 2), concentration at maximum therapeutic effect (C max ), area under the dose-response curve (AUC), and F, and quantitatively in terms of reduced clearance, dose, and material cost.

[0045] The following is intended to illustrate the above: A compound having multiple potential attack sites for oxidative metabolism, such as benzylic hydrogen atoms and hydrogen atoms attached to nitrogen atoms, is prepared as a series of analogs in which various combinations of hydrogen atoms are replaced by deuterium atoms, so that some, most, or all of these hydrogen atoms are replaced by deuterium atoms. Determination of the half-life allows for convenient and accurate determination of the extent to which the resistance to oxidative metabolism has been improved. In this way, it is determined that the half-life of the parent compound can be extended by up to 100% as a result of this type of deuterium-hydrogen exchange.

[0046] Deuterium-hydrogen exchange in the compounds disclosed herein can also be used to achieve favorable modification of the metabolite spectrum of the starting compound to reduce or eliminate undesired toxic metabolites. For example, if a toxic metabolite arises from oxidative carbon-hydrogen (CH) bond cleavage, it can be reasonably assumed that a deuterated analog will greatly reduce or eliminate the production of the undesired metabolite, even if the specific oxidation is not the rate-limiting step. Further information on the state of the art regarding deuterium-hydrogen exchange can be found, for example, in Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990; Reider et al., J. Org. Chem. 52, 3326-3334,1987; Foster, Adv. Drug Res. 14, 1-40,1985; Gillette et al, Biochemistry 33(10) 2927-2937,1994; and Jarman et al. Carcinogenesis 16(4), 683-688,1993.

[0047] Following their preparation, the compounds of the invention are preferably isolated and purified to obtain compositions containing 95% by weight or more ("substantially pure") and then used or formulated as described herein. In certain embodiments, the compounds of the invention are greater than 99% pure.

[0048] Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. The term "stable," as used herein, refers to compounds that have sufficient stability to permit manufacture and maintain compound integrity for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0049] The recitation of a listing of chemical groups in any definition of a variable herein includes definition of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0050] The present invention provides novel small molecule compounds, methods for their synthesis and pharmaceutical compositions, and methods thereof for treating or ameliorating various diseases or disorders.

[0051] The central feature of the present invention is that the compounds of the present invention are cancer targeting and slow release therapeutic agents, resulting in targeted and sustained delivery. The conjugates of the present invention are composed of a tissue protein binding agent, a cleavable linker and a small molecule drug. After local injection of the conjugate, the tissue protein binding agent, acting as a "molecular glue", binds to tissue proteins in solid tumors, thereby holding the conjugate in the solid tumor without leaking into the systemic circulation. The small molecule drug is then slowly released from the conjugate by cleavage of the cleavable linker. The slow but sustained release of the drug inside the solid tumor amplifies the tumor killing effect while minimizing adverse reactions, since a minimal amount of the drug is leaked into the systemic circulation. The dosing schedule can be varied according to the half-life of the conjugate.

[0052] In one aspect, the present invention generally relates to a compound having structural formula (I):

[0053] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof, During the ceremony, X is CHR or NR; R is H or alkyl.

[0054] In certain embodiments, the compound of formula (I) is

[0055] [ka] is selected from.

[0056] In another aspect, the present invention generally relates to a compound of structural formula (II):

[0057] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0058] In certain embodiments, the compound of formula (II) is

[0059] [ka] is selected from.

[0060] In yet another aspect, the invention generally relates to a compound having the structural formula:

[0061] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0062] In yet another aspect, the present invention generally relates to a compound having structural formula (III):

[0063] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof, In the formula, n is an integer selected from 2 to 8 (i.e., 2, 3, 4, 5, 6, 7, or 8).

[0064] In certain embodiments, the compound of formula (III) is

[0065] [ka] is selected from.

[0066] In yet another aspect, the present invention generally relates to a compound having structural formula (IV):

[0067] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof, In the formula, n is an integer selected from 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10).

[0068] In certain embodiments, the compound of formula (IV) is

[0069] [ka]

[0070] [ka]

[0071] [ka] is selected from.

[0072] In yet another aspect, the present invention generally relates to a compound having structural formula (V):

[0073] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof, During the ceremony, R 2 is H or (=O)(R 3 )2, and R 3is alkyl, L is a single bond, or

[0074] [ka] is a group selected from k is an integer selected from 0 to 4 (eg, 0, 1, 2, 3, or 4).

[0075] In certain embodiments, the compound of formula (V) is

[0076] [ka] is selected from.

[0077] In yet another aspect, the present invention generally relates to a compound having structural formula (VI):

[0078] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof, During the ceremony, R 2 is H or (=O)(R 3 )2, and R 3 is alkyl, L is a single bond, or

[0079] [ka] is a group selected from k is an integer selected from 0 to 4 (eg, 0, 1, 2, 3, or 4).

[0080] In certain embodiments, the compound of formula (VI) has the following structure:

[0081] [ka] has.

[0082] In yet another aspect, the present invention generally relates to a compound having the following structural formula:

[0083] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0084] In yet another aspect, the present invention generally relates to a compound having the following structural formula:

[0085] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0086] In yet another aspect, the present invention generally relates to a compound having the following structural formula:

[0087] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0088] In yet another aspect, the present invention generally relates to a compound having the following structural formula:

[0089] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0090] In yet another aspect, the present invention generally relates to a compound having the following structural formula:

[0091] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0092] In yet another aspect, the present invention generally relates to a compound having the following structural formula:

[0093] [ka] or a pharma- ceutically acceptable form or isotopic derivative thereof.

[0094] In yet another aspect, the invention generally relates to pharmaceutical compositions comprising a compound disclosed herein and a pharma- ceutically acceptable excipient, carrier, or diluent.

[0095] In certain embodiments, the pharmaceutical compositions of the present invention are effective in treating or reducing cancer or a related disease or disorder.

[0096] In yet another aspect, the invention generally relates to unit dosage forms that include pharmaceutical compositions that include the compounds disclosed herein.

[0097] In yet another aspect, the invention generally relates to a method of treating or reducing a disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein and a pharma- ceutical acceptable excipient, carrier, or diluent.

[0098] In certain embodiments, the disease or condition is cancer or a related disease or condition.

[0099] In yet another aspect, the invention generally relates to the use of the compounds disclosed herein for the treatment or amelioration of a disease or disorder, such as cancer.

[0100] In yet another aspect, the invention generally relates to the use of a compound disclosed herein, and a pharma- ceutically acceptable excipient, carrier, or diluent, in the preparation of a medicament for the treatment or amelioration of a disease or disorder, such as cancer.

[0101] The compositions of the present invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions of the present invention include aqueous or oily suspensions. These suspensions are formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparations are also sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, for example, solutions in 1,3-butanediol. Among the acceptable vehicles and solvents used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally used as solvents or suspending media.

[0102] For this purpose, any non-irritating fixed oil can be used, including synthetic mono- or di-glycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful for the preparation of injections, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated versions. These oil solutions or suspensions also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose, or similar dispersants that are commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability enhancers, that are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, can also be used for formulation purposes.

[0103] The pharmaceutically acceptable composition of the present invention is orally administered in any orally acceptable dosage form.Exemplary oral dosage forms are capsules, tablets, aqueous suspensions, or liquids.For tablets for oral use, commonly used carriers include lactose and corn starch.Lubricants, such as magnesium stearate, are also typically added.For oral administration in capsule form, useful diluents include lactose and dried corn starch.When aqueous suspension is required for oral use, active ingredient is combined with emulsifying and suspending agents.If desired, certain sweeteners, flavors, or colorants are also optionally added.

[0104] Alternatively, the pharma- ceutically acceptable compositions of the present invention are administered in the form of suppositories for rectal administration. These can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycols.

[0105] The pharma- ceutically acceptable compositions of the present invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0106] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.

[0107] For topical application, the provided pharma- ceutically acceptable composition is formulated into a suitable ointment, containing the active ingredient suspended or dissolved in one or more carriers.Exemplary carriers for topical administration of the compound are mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water.Alternatively, the provided pharma-ceutically acceptable composition can be formulated into a suitable lotion or cream, containing the active ingredient suspended or dissolved in one or more pharma-ceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0108] The pharma- ceutically acceptable compositions of the present invention are optionally administered by nasal aerosol or inhalation.Such compositions are prepared according to techniques well known in the art of pharmaceutical formulations, and are prepared as solutions in saline using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0109] Most preferably, the pharma- ceutically acceptable composition of the present invention is formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharma- ceutically acceptable composition of the present invention is administered without food. In other embodiments, the pharma- ceutically acceptable composition of the present invention is administered with food.

[0110] The amount of the compounds of the present invention, optionally combined with a carrier material to produce a composition in a single dosage form, will vary depending on the host being treated, the particular mode of administration. Preferably, the compositions provided should be formulated so that a dosage of 0.01-100 mg / kg body weight / day of the compound can be administered to a patient receiving these compositions.

[0111] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the particular compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, as well as the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the invention in a composition will also depend on the particular compound in the composition. EXAMPLES

[0112] As depicted in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures: While this general method depicts the synthesis of specific compounds of the invention, it will be understood that the following general method, as well as other methods known to those of skill in the art, are applicable to any compound as described herein, and each subclass and species of these compounds.

[0113] The compound numbers utilized in the following examples correspond to the compound numbers above.

[0114] 1 H was recorded at 400 MHz on a Varian Mercury400 spectrometer. 13 C NMR was recorded at 100 MHz. Proton chemical shifts were internally referenced to the residual proton resonance in CDCl3 (7.26 ppm). Carbon chemical shifts were internally referenced to the deuterated solvent signal in CDCl3 (77.20 ppm).

[0115] LC-MS spectra were recorded on a Shimadzu LC-MS2020 using an Agilent C18 column (Eclipse XDB-C18, 5um, 2.1x50mm) at a flow rate of 1mL / min. Mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile. A general gradient method was used.

[0116] [Table 1]

[0117] Analytical HPLC was performed on an Agilent 1200 HPLC using a Zorbax Eclipse XDB C18 column (2.1×150 mm) at a flow rate of 1 mL / min. Mobile phase A: 0.1% TFA in water, Mobile phase B: 0.1% TFA in acetonitrile. The general method with the following gradient was used:

[0118] [Table 2]

[0119] Preparative HPLC was performed on a Varian ProStar using a Hamilton C18 PRP-1 column (15×250 mm) at a flow rate of 20 mL / min. Mobile phase A: 0.1% TFA in water, Mobile phase B: 0.1% TFA in acetonitrile. A typical gradient method was used.

[0120] [Table 3]

[0121] Example 1. (R)-4,11-diethyl-4-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl 4-(4-iodophenyl)butanoate (1)

[0122] [ka]

[0123] [ka]

[0124] Step 1: To a solution of compound 1-2 (600 mg, 2.06 mmol) in DMF (10 mL) was added EDCI (800 mg, 4.18 mmol) and HOBt (190 mg, 1.40 mmol). After stirring for 5 min, compound 1-1 (SN-38, 500 mg, 1.27 mmol) was added and the mixture was stirred at room temperature for 16 h. After completion of the reaction, water (100 mL) was added and the mixture was stirred at room temperature for 30 min. A white precipitate was completely formed. The resulting mixture was filtered and washed with water to obtain an off-white solid. The crude product was then triturated with acetonitrile for 1 h and filtered. The filter cake was washed with acetonitrile and dried to obtain the title compound 1 (505 mg, 59% yield) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ8.19(d,J=9.1Hz,1H),8.04-7.94(m,1H),7.66(td,J= 10.1,9.2,4.6Hz,3H),7.32(s,1H),7.10(d,J=7.8Hz,2H),6.53(s,1H),5.44( s,2H),5.32(s,2H),3.17(q,J=7.6Hz,2H),2.76-2.62(m,4H),1.98(p,J=7.4H z,2H),1.87(hept,J=7.0Hz,2H),1.28(t,J=7.5Hz,3H),0.88(t,J=7.3Hz,3H). LCMS:C 32 H 29 Calculated m / z for IN2O6: 664.50, Found: 665.2 [M+H] + .

[0125] Example 2. (R)-4,11-diethyl-4-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl 2-methyl-4-(4-iodophenyl)butanoate (2)

[0126] [ka]

[0127] [ka]

[0128] Step 1: LDA (2M, 2.5 mL, 5 mmol) was added dropwise to a stirred solution of compound 1-2 (500 mg, 1.72 mmol) in anhydrous THF (10 mL) at 0° C. under nitrogen atmosphere. After addition, the mixture was stirred at 0° C. for 30 min. MeI (538 mg, 3.79 mmol) was added dropwise and the reaction was stirred at room temperature for 16 h. After completion of the reaction, the mixture was quenched with saturated NH4Cl (30 mL) solution and extracted with EtOAc (30 mL×3). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 2-1 (400 mg, 76% yield).

[0129] Step 2: A mixture of compound 2-1 (390 mg, 1.24 mmol), compound 1-1 (194 mg, 0.496 mmol), HOBt (335 mg, 2.48 mmol), DMAP (30 mg, 0.248 mmol), and EDCI (950 mg, 4.97 mmol) in DMF (5 mL) and DCM (5 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 5% MeOH in DCM) to give the title compound 2 (265 mg, 30% yield).

[0130] Example 3. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl(4-iodophenethyl)carbamate (3)

[0131] [ka]

[0132] [ka]

[0133] Step 1: To a solution of compound 3-1 (1.0 g, 4.11 mmol) in anhydrous THF (10 mL) at 0° C., a solution of borane-tetrahydrofuran complex (1 M in THF, 18.5 mL) was added dropwise under nitrogen atmosphere. The reaction mixture was stirred at 65° C. for 2 h. After completion of the reaction, the mixture was cooled to 0° C. and quenched by the addition of 6N HCl (2 mL). The resulting mixture was then basified with 1N NaOH solution and extracted with DCM (50 mL×2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give the title compound 3-2 (1.0 g, 90% yield) as a colorless oil, which was used directly in the next step without further purification.

[0134] Step 2: Compound 3-3 (345 mg, 1.71 mmol) was dissolved in DCM (5 mL) and cooled to 0° C. To this was slowly added a solution of compound 3-2 (210 mg, 0.85 mmol) and TEA (257 mg, 2.55 mmol) in DMSO (2 mL). The mixture was stirred at 0° C. for 1 h. SN-38 (1-1, 330 mg, 0.84 mmol) and TEA (100 mg, 0.99 mmol) were added. The mixture was then stirred at room temperature for 3 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 5% MeOH in DCM) to give the title compound 3 (120 mg, 20% yield).

[0135] Example 4. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl(4-iodophenethyl)(methyl)carbamate (4)

[0136] [ka]

[0137] [ka]

[0138] Step 1: A mixture of compound 3-2 (1.0 g, 4.04 mmol), Boc2O (1.7 g, 7.79 mmol) and TEA (830 mg, 8.20 mmol) in DCM (10 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated. The residue was purified by silica gel column (eluted with 10% EtOAc in petroleum ether) to give the title compound 4-1 (1.1 g, 80% yield) as a white solid. LCMS: C 13 H 18 Calculated m / z for INO2: 347.20, Measured: 348.21 [M+H] + .

[0139] Step 2: NaH (60% in mineral oil, 450 mg, 11.25 mmol) was added in portions to a solution of compound 4-1 (1.3 g, 3.74 mmol) in anhydrous DMF (5 mL) at 0° C. After stirring for 1 h, MeI (1.6 g, 11.27 mmol) was added to the mixture in one portion. The mixture was then stirred at room temperature for 16 h. After completion of the reaction, the mixture was quenched by addition of ice water and partitioned between EtOAc (100 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by silica gel column (eluted with 5% EtOAc in petroleum ether) to give the title compound 4-2 (1.02 g, 75% yield) as a white solid. LCMS: C 14 H 20 Calculated m / z for INO2: 361.22, observed: 306.04 [M- t Bu+H] + .

[0140] Step 3: A mixture of compound 4-2 (1.02 g, 2.82 mmol) in EtOAc (5 mL) and a solution of HCl in EtOAc (4N, 5 mL) was stirred at room temperature for 1 h. After completion of the reaction, a white precipitate was completely formed. The white precipitate was filtered and washed with EtOAc to give the title compound 4-3 (610 mg, 82% yield), which was used directly in the next step without further purification.

[0141] Step 4: A suspension of SN-38 (1-1, 250 mg, 0.637 mmol), DIPEA (170 mg, 1.31 mmol), and compound 4-4 (215 mg, 0.706 mmol) in anhydrous DMF (5 mL) was stirred at 0° C. for 1 h under nitrogen atmosphere. The mixture was then treated with a solution of compound 4-3 (227 mg, 0.763 mmol) and DIPEA (200 mg, 1.55 mmol). After stirring for another 2 h. The resulting mixture was diluted with EtOAc (50 mL) and washed with water. The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to give 4 (170 mg, 39% yield) as a pale yellow solid. 1 H NMR (400MHz, chloroform-d) δ8.37-8.16(m,1H),7.84(d,J=2.8Hz,1H),7.69(dd,J=5.6,3.0Hz,3H),7.61-7.48(m,1 H),7.35-7.28(m,1H),7.16-6.97(m,2H),5.79(dd,J=16.3,2.8Hz,1H),5.49-5.26(m,3H),3.91(s,1H),3.80(t ,J=7.1Hz,1H),3.66(t,J=7.8Hz,1H),3.53(d,J=2.8Hz,1H),3.19(d,J=7.4Hz,1H),3.16(d,J=2.8Hz,1H),3.09 (d,J=2.8Hz,1H),3.01-2.91(m,2H),1.93(ddd,J=15.0,7.4,4.0Hz,2H),1.48-1.38(m,3H),1.13-1.02(m,3H). LCMS:C 32 H 30 Calculated m / z for IN3O6: 679.51, Found: 680.31 [M+H]+ .

[0142] Example 5. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl 2-(4-isobutylphenyl)propanoate (5)

[0143] [ka]

[0144] [ka]

[0145] Step 1: A mixture of compound 5-1 (236.6 mg, 1.15 mmol), EDCI (438.1 mg, 2.29 mmol), and HOBt (103.2 mg, 0.76 mmol) in DMF (3 mL) was stirred at room temperature for 5 min. Compound 1-1 (300 mg, 0.76 mmol) was added and the mixture was stirred at room temperature for 15 h. The mixture was partitioned between EtOAc (30 mL) and water (30 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to give the title compound 5 (324 mg, 72% yield) as a white solid. 1 H NMR(500MHz,DMSO-d6)δ8.18(d,J=9.1Hz,1H),7.89(d,J=2.4Hz,1H),7.50(dd,J=9.2,2 .5Hz,1H),7.39-7.36(m,2H),7.32(s,1H),7.22-7.19(m,2H),6.53(s,1H),5.43(s,2H) ,5.31(s,2H),4.15(q,J=7.1Hz,1H),3.15(q,J=7.6Hz,2H),2.46(d,J=7.2Hz,2H),1.92 -1.81(m,3H),1.57(d,J=7.1Hz,3H),1.27(t,J=7.6Hz,3H),0.88(dd,J=7.1,2.3Hz,9H). LCMS:C35 H 36 Calculated m / z for N2O6: 580.68, Found: 581.47 [M+H] + .

[0146] Example 6. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl (S)-2-(6-methoxynaphthalen-2-yl)propanoate (6)

[0147] [ka]

[0148] [ka]

[0149] Step 1: A mixture of compound 6-1 (176.0 mg, 0.76 mmol), SN-38 (1-1) (150 mg, 0.38 mmol), HOBt (51.6 mg, 0.38 mmol), and EDCI (219.0 mg, 1.15 mmol) in DMF (3 mL) was stirred at room temperature for 9 h. After completion of the reaction, the reaction mixture was diluted with EtOAc and washed with water. The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column (eluted with 2% MeOH in dichloromethane) to give 6 (185 mg, 80% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ8.16(d,J=9.1Hz,1H),7.91(dd,J=4.4,2.1Hz,2H),7.87(d,J=8.7Hz,2H),7.58( dd,J=8.5,1.8Hz,1H),7.52(dd,J=9.1,2.5Hz,1H),7.34(d,J=2.6Hz,1H),7.30(s,1H),7.19(dd,J=9.0, 2.5Hz,1H),6.53(s,1H),5.43(s,2H),5.30(s,2H),4.32(q,J=7.1Hz,1H),3.87(s,3H),3.14(q,J=7.5Hz ,2H),1.85(dq,J=14.2,7.1Hz,2H),1.66(d,J=7.1Hz,3H),1.25(t,J=7.6Hz,3H),0.87(t,J=7.3Hz,3H). LCMS:C 36 H 32 Calculated m / z for N2O7: 604.66, Found: 605.36. [M+H] + TLC(DCM:MeOH / 40:1):R f (Compound 6-1)=0.5, R f (Compound 1-1)=0.3, R f (6)=0.37.

[0150] Example 7. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl (R)-2-(4-isobutylphenyl)propanoate (7)

[0151] [ka]

[0152] [ka]

[0153] Step 1: A mixture of compound 7-1 (107 mg, 0.51 mmol), EDCI (198.2 mg, 1.04 mmol), and HOBt (46.7 mg, 0.35 mmol) in DMF (3 mL) was stirred at room temperature for 5 min. Compound 1-1 (135.7 mg, 0.35 mmol) was added, and the mixture was stirred at room temperature for 15 h. The reaction mixture was partitioned between EtOAc (20 mL) and water (20 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to give the title compound 7 (161 mg, 80% yield) as a white solid. 1 H NMR(500MHz,DMSO-d6)δ8.22-8.11(m,1H),7.93-7.86(m,1H),7.53-7.46(m,1H),7.38( d,J=7.6Hz,2H),7.32(d,J=2.3Hz,1H),7.21(d,J=7.8Hz,2H),6.53(s,1H),5.44(s,2H) ,5.33(s,2H),4.16(q,J=7.2Hz,1H),3.16(p,J=6.6,6.1Hz,2H),2.47(d,J=7.2Hz,2H), 1.93-1.80(m,3H),1.57(d,J=7.1Hz,3H),1.27(t,J=7.6Hz,3H),0.88(d,J=6.3Hz,9H). LCMS:C 35 H 36 Calculated m / z for N2O6: 580.68, Found: 581.47 [M+H] + .

[0154] Example 8. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl (S)-2-(4-isobutylphenyl)propanoate (8)

[0155] [ka]

[0156] [ka]

[0157] Step 1: A mixture of compound 8-1 (157.7 mg, 0.76 mmol), EDCI (292.0 mg, 1.53 mmol), and HOBt (68.8 mg, 0.51 mmol) in DMF (2 mL) was stirred at room temperature for 5 min. Compound 1-1 (200 mg, 0.51 mmol) was added, and the mixture was stirred at room temperature for 15 h. The reaction mixture was partitioned between EtOAc (30 mL) and water (30 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to give the title compound 8 (197 mg, 66% yield) as a white solid. 1 H NMR(500MHz,DMSO-d6)δ8.17(d,J=9.1Hz,1H),7.89(t,J=2.3Hz,1H),7.50(dd,J=9.1,2 .5Hz,1H),7.39-7.36(m,2H),7.31(s,1H),7.22-7.19(m,2H),6.52(s,1H),5.43(s,2H) ,5.31(s,2H),4.15(q,J=7.1Hz,1H),3.15(q,J=7.6Hz,2H),2.46(d,J=7.1Hz,2H),1.92 -1.80(m,3H),1.57(d,J=7.1Hz,3H),1.26(t,J=7.6Hz,3H),0.87(dd,J=7.0,2.8Hz,9H). LCMS:C 35 H 36 Calculated m / z for N2O6: 580.68, Found: 581.47 [M+H] + .

[0158] Example 9. N-((S)-1-(((R)-1-(6-(2-(4-(4-iodophenyl)butanamido)ethyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (9)

[0159] [ka]

[0160] [ka]

[0161] Step 1: To a solution of compound 9-1 (1.0 g, 6.24 mmol) in DMF (25 mL) was added ethyl bromide (9-2, 9.9 g, 59.2 mmol), KI (1.04 g, 6.26 mmol), and NaHCO3 (5.0 g, 59.5 mmol). The mixture was stirred at room temperature for 14 h. After completion of the reaction, the resulting mixture was diluted with EtOAc (200 mL) and washed with water (200 mL x 2) and brine (200 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (eluted with 20% EtOAc in petroleum ether) to give the title compound 9-3 (1.7 g, 81% yield) as a colorless oil.

[0162] Step 2: A mixture of compound 9-3 (500 mg, 1.50 mmol) and TFA (1 mL) in DCM (4 mL) was stirred at room temperature for 1 h. After completion of the reaction, the resulting mixture was concentrated to give title compound 9-4 (600 mg) as a colorless oil, which was used in the next step without further purification.

[0163] Step 3: Compound 9-4 (600 mg, crude from step 2) was dissolved in DCM (5 mL) and adjusted to pH=8 by the addition of Et3N. The solution was then added to a stirred mixture of compound 1-2 (360 mg, 1.24 mmol) and EDCI (431 mg, 2.25 mL) in DCM (10 mL). After stirring for 16 h, the resulting mixture was diluted with DCM (50 mL) and washed with water (50 mL) and 1M HCl (50 mL×2). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 9-5 (600 mg,) as a colorless oil, which was used in the next step without further purification.

[0164] Step 4: A mixture of compound 9-5 (600 mg, crude from step 3) in MeOH (10 mL) was treated with aqueous NaOH (1 M, 3.67 mL, 3.67 mmol) and stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to remove the organic solvent. The aqueous solution was washed with EtOAc (50 mL x 3) and acidified to pH = 2 with 1 M HCl to form a white precipitate. The precipitate was filtered and washed with water to give the title compound 9-6 (180 mg, 26% yield for three steps) as a white solid. LCMS: C 16 H 21 Calculated m / z for IN2O5: 448.26, Found: 449.03 [M+H] + .

[0165] Step 5: A mixture of compound 9-6 (180 mg, 0.40 mmol) and compound 9-7 (155 mg, 0.40 mmol) in toluene (18 mL) was stirred at 120° C. for 3 h. After completion of the reaction, the mixture was concentrated. The residue was washed with MTBE and filtered to obtain a solid. The crude product was redissolved in acetonitrile and purified by pre-TLC (SiO2, DCM / acetonitrile=2 / 1) to give the title compound 9 (50.1 mg, 15% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ9.09(s,1H),8.90(d,J=2.5Hz,1H),8.83(d,J=8.6Hz,1H),8.75(s,1H),8.17(s,1H),7.62(d,J=7.8Hz,2H),7 .42(d,J=9.7Hz,1H),7.28(q,J=8.1,7.4Hz,4H),7.20(d,J=7.0Hz,1H),7.02(d,J=7.9Hz,2H),4.74(q,J=8.2Hz,1H),4.33(d,J=17.2 Hz,1H),4.24-4.11(m,2H),3.79(d,J=16.8Hz,1H),3.62-3.53(m,1H),3.22(s,1H),3.17(s,1H),3.06(d,J=8.5Hz,2H),2.51(s,1H), 2.17-2.09(m,2H),1.80(t,J=7.7Hz,2H),1.59(s,1H),1.42(t,J=12.9Hz,1H),1.25(d,J=14.1Hz,4H),0.86(dd,J=13.0,6.5Hz,6H). LCMS:C 35 H 42 Calculated m / z for BIN6O7: 796.47, Found: 797.6 [M+H] + .

[0166] Example 10. N-((S)-1-(((R)-1-(6-(4-(4-(4-iodophenyl)butanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (10)

[0167] [ka]

[0168] [ka]

[0169] Step 1: A solution of compound 1-2 (400 mg, 1.38 mmol), compound 10-1 (312 mg, 1.66 mmol), DMAP (17 mg, 0.14 mmol), and EDCI (397 mg, 2.07 mmol) in DCM (20 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was diluted with DCM (100 mL) and washed with 1N HCl and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 10-2 (500 mg), which was used in the next step without further purification.

[0170] Step 2: Compound 10-2 (500 mg, crude from step 1) was dissolved in 4N HCl in EtOAc (5 mL) and the solution was stirred for 1 h. After completion of the reaction, the mixture was concentrated to give the title compound 10-3 (450 mg), which was used in the next step without further purification.

[0171] Step 3: A mixture of compound 10-3 (450 mg, crude from step 2), compound 10-4 (600 uL, 4.14 mmol), and triethylamine (1.0 mL, 6.90 mmol) in acetonitrile (10 mL) was stirred at room temperature for 18 h. After completion of the reaction, the reaction mixture was diluted with DCM (100 mL) and washed with water and brine. The organic layer was dried over Na2SO4, filtered, and concentrated to give the title compound 10-5 (300 mg), which was used in the next step without further purification.

[0172] Step 4: Compound 10-5 (300 mg, crude from step 3) was dissolved in 4N HCl in EtOAc (5 mL) and stirred for 2 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by preparative HPLC (C18 column, eluted with acetonitrile and H2O, TFA condition) to give the title compound 10-6 (110 mg, yield: 13.4% for 4 steps) as a pale yellow oil.

[0173] Step 5: A mixture of compound 10-6 (112 mg, 0.235 mmol) and compound 9-7 (90 mg, 0.234 mmol) in toluene (5 mL) was stirred at 120° C. for 3 h. After completion of the reaction, the mixture was concentrated. The residue was washed with MTBE and filtered to obtain a solid. The crude product was redissolved in acetonitrile and purified by pre-TLC (SiO2, DCM / acetonitrile=3 / 1) to give the title compound 10 (51 mg, 26% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ9.07(s,1H),8.89(d,J=8.3Hz,1H),8.86(d,J=2.3Hz,1H),8.73(s,1H),7.88-7.79(m,1H),7.59(dd,J= 8.2,1.9Hz,2H),7.34(d,J=9.6Hz,1H),7.30-7.20(m,4H),7.19-7.12(m,1H),7.03-6.95(m,2H),4.67(q,J=7.7Hz,1H),4.17(d ,J=17.3Hz,1H),4.05(d,J=17.4Hz,1H),3.95(d,J=16.6Hz,1H),3.63-3.45(m,2H),3.07(d,J=7.3Hz,6H),2.09-2.03(m,2H),1 .77(p,J=7.4Hz,2H),1.56(s,3H),1.41(t,J=12.6Hz,3H),1.21(d,J=17.2Hz,2H),0.85(d,J=6.6Hz,3H),0.80(d,J=6.3Hz,3H). LCMS:C 37 H 46 Calculated m / z for BIN6O7: 824.52, Found: 825.9. [M+H] + .

[0174] Example 11. N-((S)1-(((R)1-(6-(3-(4-(4-iodophenyl)butanamido)propyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (11)

[0175] [ka]

[0176] [ka]

[0177] Step 1: A mixture of compound 11-1 (1.0 g, 6.79 mmol), NaHCO3 (4.8 g, 57.07 mmol), KI (950 mg, 5.72 mmol), and compound 9-2 (9.5 g, 56.89 mmol) in DMF (20 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by silica gel column (eluted with 15% EtOAc in petroleum ether) to give the title compound 11-2 (1.7 g, 85% yield) as a colorless oil. TLC (DCM:MeOH / 10:1):R f (Compound 11-1) = 0.2, TLC (petroleum ether: EtOAc / 2:1): R f (Compound 11-2)=0.45.

[0178] Step 2: A solution of compound 11-2 (700 mg, 2.02 mmol) and TFA (4 mL) in DCM (4 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated to give the title compound 11-3 (680 mg, 70% yield) as a colorless oil, which was used in the next step without further purification. TLC (petroleum ether: EtOAc / 2:1): R f (Compound 11-2)=0.45, TLC(DCM:MeOH / 5:1):R f (Compound 11-3)=0.15.

[0179] Step 3: A mixture of crude compound 11-3 (680 mg, 1.43 mmol), compound 1-1 (527 mg, 1.82 mmol), EDCI (580 mg, 3.04 mmol), and DMAP (25 mg, 0.205 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (20 mL). The organic layer was washed with 1N HCl (50 mL×2), saturated Na2CO3 solution, and brine (50 mL), dried over anhydrous Na2SO4, and concentrated to give the title compound 11-4 (580 mg, 78% yield) as a colorless oil, which was used directly in the next step without further purification. LCMS: C 21 H 31 Calculated m / z for IN2O5: 518.39, Found: 519.09 [M+H] + .

[0180] Step 4: A mixture of compound 11-4 (580 mg, 1.12 mmol) and NaOH (290 mg, 7.25 mmol) in EtOH (10 mL) and HO (5 mL) was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated to remove the organic solvent. The remaining aqueous solution was adjusted to pH=2 and purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and water, TFA condition) to give the title compound 11-5 (220 mg, 43% yield) as a white solid. LCMS: C 17 H 23 Calculated m / z for IN2O5: 462.28, Found: 463.51 [M+H] + .

[0181] Step 5: A mixture of compound 11-5 (120 mg, 0.259 mmol) and compound 9-7 (83 mg, 0.216 mmol) in toluene (5 mL) was stirred at 120° C. for 13 h. After completion of the reaction, the mixture was concentrated. The residue was purified by pre-TLC (DCM:acetonitrile=3:1) to give 11 (63 mg, 35% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ9.07(s,1H),8.92-8.83(m,2H),8.77-8.70(m,1H),7.74(s,1H),7.63-7.54(m,2H),7.3 8(d,J=9.8Hz,1H),7.27(dd,J=13.3,7.8Hz,4H),7.19-7.14(m,1H),6.98(d,J=7.7Hz,2H),4.67(d,J=8.4Hz,1H) ,4.17-4.05(m,2H),3.98(d,J=16.6Hz,1H),3.53(d,J=22.0Hz,2H),3.07(d,J=11.9Hz,6H),2.08(d,J=6.9Hz,2H ),1.80-1.70(m,3H),1.57(s,2H),1.42(t,J=13.1Hz,2H),1.21(d,J=18.3Hz,2H),0.82(dd,J=22.8,6.3Hz,6H). LCMS:C 36 H 44 Calculated m / z for BIN6O7: 810.50, Found: 811.63 [M+H] + .

[0182] Example 12. N-((S)-1-(((R)-1-(6-(5-(4-(4-iodophenyl)butanamido)pentyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (12)

[0183] [ka]

[0184] [ka]

[0185] Step 1: A mixture of compound 12-1 (500 g, 2.47 mmol), NaHCO3 (2.1 g, 24.9 mmol), KI (410 mg, 2.47 mmol), and compound 9-2 (4.1 mg, 24.5 mmol) in DMF (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by silica gel column (eluted with 15% EtOAc in petroleum ether) to give the title compound 12-2 (700 mg, 75% yield) as a colorless oil. TLC (DCM:MeOH / 10:1):R f (Compound 12-1) = 0.2, TLC (petroleum ether: EtOAc / 2:1): R f (Compound 12-2)=0.55.

[0186] Step 2: A solution of compound 12-2 (700 mg, 1.87 mmol) and TFA (4 mL) in DCM (4 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated to give the title compound 12-3 (760 mg, 80% yield) as a colorless oil, which was used in the next step without further purification. TLC (petroleum ether: EtOAc / 2:1): R f (Compound 12-2)=0.55, TLC(DCM:MeOH / 5:1):R f (Compound 12-3)=0.18.

[0187] Step 3: A mixture of crude compound 12-3 (660 mg, 1.51 mmol), compound 1-2 (488 mg, 1.68 mmol), EDCI (540 mg, 2.82 mmol), and DMAP (23 mg, 0.189 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (20 mL). The organic layer was washed with 1N HCl (50 mL×2), saturated Na2CO3 solution, and brine (50 mL), dried over anhydrous Na2SO4, and concentrated to give the title compound 12-4 (700 mg, 84% yield) as a colorless oil, which was used in the next step without further purification. TLC (petroleum ether:EtOAc / 2:1): R f (Compound 12-3)=0.5, R f (Compound 12-4)=0.2.

[0188] Step 4: A mixture of compound 12-4 (700 mg, 1.28 mmol) and NaOH (270 mg, 6.75 mmol) in EtOH (10 mL) and HO (5 mL) was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated to remove the organic solvent. The remaining aqueous solution was adjusted to pH=2 and directly purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and water, TFA condition) to give the title compound 12-5 (270 mg, 43% yield) as a white solid. LCMS: C 19 H 27 Calculated m / z for IN2O5: 490.34, Found: 491.30 [M+H] + .

[0189] Step 5: A mixture of compound 12-5 (153 mg, 0.312 mmol) and compound 9-7 (100 mg, 0.260 mmol) in toluene (5 mL) was stirred at 120° C. for 13 h. After completion of the reaction, the mixture was concentrated. The residue was purified by pre-TLC (DCM:acetonitrile=3:1) to give 12 (67 mg, 25% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ9.05(d,J=3.0Hz,1H),8.94-8.82(m,2H),8.71(d,J=3.3Hz,1H),7.81(s,1H),7.56(dd,J=8.1,2.8 Hz,2H),7.35(d,J=10.1Hz,2H),7.30-7.21(m,4H),7.19-7.13(m,1H),6.96(dd,J=8.4,2.8Hz,2H),4.66(s,1H),4.15(dd, J=17.5,3.0Hz,1H),4.07-3.99(m,1H),3.93(dd,J=16.8,3.0Hz,1H),3.05(d,J=8.4Hz,6H),2.03(d,J=8.1Hz,2H),1.75(d ,J=8.0Hz,2H),1.54(s,4H),1.41(d,J=9.9Hz,3H),1.15(s,4H),0.83(dd,J=6.6,3.1Hz,3H),0.79(dd,J=6.6,3.1Hz,3H). LCMS:C 38 H 48 Calculated m / z for BIN6O7: 838.55, Found: 839.50 [M+H] + .

[0190] Example 13. N-((2S)-1-(((1R)-1-(6-(4-(2-(4-isobutylphenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (13)

[0191] [ka]

[0192] [ka]

[0193] Step 1: A mixture of compound 10-1 (2 g, 10.62 mmol), compound 9-2 (17.7 g, 106.23 mmol), NaHCO3 (8.92 g, 106.23 mmol), and KI (1.76 g, 10.62 mmol) in DMF (20 mL) was stirred at room temperature for 16 h. The reaction mixture was diluted with EtOAc (100 mL) and washed with water (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 13-1 (3.5 g, 91% yield) as a yellow oil.

[0194] Step 2: To a solution of compound 13-1 (2 g, 5.55 mmol) in DCM (16 mL) was added TFA (4 mL) at room temperature and the mixture was stirred for 1 h. After completion of the reaction, the mixture was concentrated to give crude compound 13-2 (3 g), which was used directly in the next step.

[0195] Step 3: A mixture of compound 5-1 (1.14 g, 5.53 mmol), HATU (3.15 g, 8.29 mmol), and DIPEA (1.37 mL, 8.29 mmol) in DMF (2 mL) was stirred at room temperature for 10 min. Compound 13-2 (3 g, crude from step 2) was added and the mixture was stirred for 3 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and H2O, HCl condition) to give the title compound 13-3 (1.8 g, 72% yield, 2 steps). LCMS: C 25 H 40 Calculated m / z for N2O5: 448.29, Found: 473.32 [M+H] + .

[0196] Step 4: To a mixture of compound 13-3 (1.8 g, 4.01 mmol) in EtOH (8 mL) and H2O (4 mL), LiOH (336.7 mg, 20.06 mmol) was added. The mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was adjusted to pH=3 with 1N HCl and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and H2O, HCl condition) to give the title compound 13-4 (1.1 g, 70% yield). LCMS: C 21 H 32 Calculated m / z for N2O5: 392.50, Found: 393.33 [M+H] + .

[0197] Step 5: A mixture of compound 13-4 (1 g, 0.36 mmol) and compound 9-7 (115 mg, 0.30 mmol) in toluene (15 mL) was stirred at 120° C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by silica gel column (eluted with 30% acetonitrile in DCM) to give the title compound 13 (1 g, 53% yield) as a pale yellow solid. 1H NMR(500MHz,DMSO-d6)δ9.09(dd,J=3.8,1.5Hz,1H),8.92-8.85(m,2H),8.73(ddd,J=5.1,2.5,1.5Hz,1H),7.97(td,J=5.8,2.1Hz,1H),7.35(dd, J=9.9,6.0Hz,1H),7.31-7.15(m,7H),7.06(d,J=7.9Hz,2H),4.73-4.63(m,1H),4.12(dd,J=17.4,3.6Hz,1H),4.03(dd,J=17.3,1.7Hz,1H),3.98- 3.89(m,1H),3.56(qd,J=8.1,7.0,3.5Hz,2H),3.48(dd,J=16.7,8.7Hz,1 H),3.15-2.97(m,6H),2.38(d,J=7.2Hz,2H),1.78(dp,J=13.5,6.7Hz,1H ),1.62-1.50(m,2H),1.50-1.40(m,2H),1.40-1.33(m,2H),1.32(dd,J=7 .0,1.4Hz,3H),1.21(ddt,J=13.7,10.2,4.5Hz,1H),0.92-0.74(m,12H). LCMS:C 40 H 53 Calculated m / z for BN6O7: 740.41, Found: 742.01 [M+H] + .

[0198] Example 14. N-((S)-1-(((R)-1-(6-(4-((S)-2-(6-methoxynaphthalen-2-yl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (14)

[0199] [ka]

[0200] [ka]

[0201] Step 1: A mixture of compound 6-1 (254 mg, 1.11 mmol), DIPEA (445 mg, 3.45 mmol), compound 13-2 (415 mg, crude, ca. 70% purity, 1.12 mmol), and HATU (787 mg, 2.07 mmol) in DMF (3 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and water, HCl condition). The desired component was lyophilized to give the title compound 14-1 (380 mg, 58% yield). LCMS: C 26 H 36 Calculated m / z for N2O6: 472.3, Found: 473.32 [M+H] + .

[0202] Step 2: A mixture of compound 14-1 (380 mg, 0.80 mmol) and NaOH (160 mg, 4.01 mmol) in EtOH (5 mL) and HO (5 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was adjusted to pH=7 with 1N HCl. The resulting mixture was concentrated and purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and water, HCl condition). The desired component was lyophilized to give the title compound 14-2 (150 mg, 45% yield). LCMS: C 26 H 36 Calculated m / z for N2O6: 416.19, Found: 417.13. [M+H] + .

[0203] Step 3: A mixture of compound 14-2 (150 mg, 0.36 mmol) and compound 9-7 (115 mg, 0.30 mmol) in toluene (3 mL) was stirred at 120° C. for 17 h. After completion of the reaction, the mixture was concentrated to remove the organic solvent. The crude product was purified by pre-HPLC (C18 column, eluted with acetonitrile and HO, neutral condition) to give the title compound 14 (30 mg, 13% yield). 1H NMR(400MHz,DMSO-d6)δ9.05(d,J=4.0Hz,1H),8.86(d,J=13.7Hz,2H),8.68(s,1H),8.05(d,J=6.4Hz,1H),7.72(d t,J=13.6,6.0Hz,3H),7.47-7.40(m,1H),7.34(d,J=9.1Hz,1H),7.28-7.19(m,5H),7.18-7.08(m,2H),4.66(s,1H) ,4.09(d,J=17.7Hz,1H),3.98(d,J=17.5Hz,1H),3.88(d,J=14.9Hz,1H),3.83(d,J=4.3Hz,3H),3.71(s,2H),3.06 (s,6H),1.53(s,2H),1.39(q,J=13.4,9.6Hz,6H),1.28-1.12(m,4H),0.83(t,J=5.2Hz,3H),0.79(t,J=5.5Hz,3H). LCMS:C 41 H 49 m / z calculated for BN6O8: 764.37, found: 765.43. [M+H] + .

[0204] Example 15. N-((S)-1-(((R)-1-(6-(4-((R)-2-(4-isobutylphenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (15)

[0205] [ka]

[0206] [ka]

[0207] Step 1: A mixture of compound 7-1 (143 mg, 0.69 mmol), HATU (396 mg, 1.04 mmol), and DIPEA (224 mg, 1.73 mmol) in DMF (2 mL) was stirred at room temperature for 10 min, and compound 13-2 (300 mg, crude, approx. 70% purity, 0.81 mmol) was added. After stirring at room temperature for 3 h, the mixture was diluted with EtOAc (20 mL) and washed with water (20 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and H2O, HCl condition) to give the title compound 15-1 (307 mg, 99% yield). LCMS: C 25 H 40 Calculated m / z for N2O5: 448.60, Found: 449.62 [M+H] + .

[0208] Step 2: To a mixture of compound 15-1 (337 mg, 0.75 mmol) in EtOH (2 mL) and H2O (2 mL), LiOH.H2O (157.6 mg, 3.76 mmol) was added. The mixture was then stirred at room temperature for 1 h. After completion of the reaction, the mixture was adjusted to pH=7 with 1N HCl and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and H2O, HCl condition) to give the title compound 15-2 (231 mg, 78% yield). LCMS: C 25 H 40 Calculated m / z for N2O5: 392.23, Found: 393.33 [M+H] + .

[0209] Step 3: A mixture of compound 15-2 (100 mg, 0.25 mmol) and compound 9-7 (81.6 mg, 0.21 mmol) in toluene (5 mL) was stirred at 120° C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (SiO2, acetonitrile / DCM=1 / 3) to give the title compound 15 (97 mg, 61% yield) as a white solid. 1H NMR(500MHz,DMSO-d6)δ9.09(d,J=1.5Hz,1H),8.92-8.86(m,2H),8.74(dd,J=2.5,1.5Hz,1H),7.97(t,J=5.8Hz,1H),7.34(d,J=9.8Hz,1H),7.31-7. 27(m,2H),7.27-7.23(m,2H),7.22-7.19(m,2H),7.19-7.15(m,1H),7.08- 7.03(m,2H),4.68(q,J=7.7Hz,1H),4.11(d,J=17.4Hz,1H),4.03(d,J=17.3 Hz,1H),3.92(d,J=16.7Hz,1H),3.56(qd,J=6.9,3.1Hz,2H),3.49(d,J=16 .8Hz,1H),3.13-2.98(m,6H),2.38(d,J=7.1Hz,2H),1.78(hept,J=6.8Hz,1 H),1.57(ddt,J=13.7,6.7,3.6Hz,2H),1.49-1.34(m,4H),1.32(d,J=7.1H z,3H),1.20(ddd,J=17.2,10.2,5.0Hz,1H),0.83(dt,J=13.6,6.9Hz,12H). LCMS:C 40 H 53 Calculated m / z for BN6O7: 740.41, Found: 741.94 [M+H] + .

[0210] Example 16. N-((S)-1-(((R)-1-(6-(4-((S)-2-(4-isobutylphenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (16)

[0211] [ka]

[0212] [ka]

[0213] Step 1: A mixture of compound 8-1 (334 mg, 1.62 mmol), HATU (924 mg, 2.42 mmol), and DIPEA (716.8 μL, 4.04 mmol) in DMF (5 mL) was stirred at room temperature for 10 min, and compound 13-2 (800 mg, crude, 70% purity, 2.15 mmol) was added. After stirring at room temperature for 3 h, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and H2O, HCl condition) to give the title compound 16-1 (706 mg, 97% yield). LCMS: C 25 H 40 Calculated m / z for N2O5: 448.60, Found: 449.62 [M+H] + .

[0214] Step 2: To a mixture of compound 16-1 (806 mg, 1.80 mmol) in EtOH (5 mL) and H2O (5 mL), LiOH.H2O (376.9 mg, 8.98 mmol) was added and the mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was adjusted to pH=7 with 1N HCl and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and H2O, HCl condition) to give the title compound 16-2 (549 mg, 78% yield). LCMS: C 25 H 40 Calculated m / z for N2O5: 392.23, Found: 393.33 [M+H] + .

[0215] Step 3: A mixture of compound 16-2 (100 mg, 0.25 mmol) and compound 9-7 (81.6 mg, 0.21 mmol) in toluene (5 mL) was stirred at 120° C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile / DCM=1 / 3) to give compound 16 (98 mg, 62% yield) as a white solid. 1H NMR(500MHz,DMSO-d6)δ9.09(d,J=1.5Hz,1H),8.90-8.85(m,2H),8.74-8.71(m,1H),7.97(t,J=5.8Hz,1H),7.36(d,J=9.8Hz,1H),7.31-7.27 (m,2H),7.27-7.23(m,2H),7.23-7.20(m,2H),7.19-7.15(m,1H),7.08 -7.04(m,2H),4.75-4.64(m,1H),4.12(d,J=17.4Hz,1H),4.03(d,J=17 .3Hz,1H),3.92(d,J=16.7Hz,1H),3.61-3.52(m,2H),3.47(d,J=16.7Hz,1H),3.14-2.98(m,6H),2.38(d,J=7.1Hz,2H),1.79(dh,J=13.6,6.8 Hz,1H),1.62-1.50(m,2H),1.50-1.34(m,4H),1.32(d,J=7.0Hz,3H),1.21(ddd,J=17.2,9.5,4.3Hz,1H),0.84(td,J=10.4,9.7,6.5Hz,12H). LCMS:C 40 H 53 Calculated m / z for BN6O7: 740.41, Found: 741.73 [M+H] + .

[0216] Example 17. N-((S)-1-(((R)-1-(6-(5-((S)-2-(4-isobutylphenyl)propanamido)pentyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (17)

[0217] [ka]

[0218] [ka]

[0219] Step 1: To a solution of compound 17-1 (1.0 g, 4.94 mmol) in DMF (10 mL), NaHCO3 (4.15 g, 49.39 mmol), KI (820 mg, 4.93 mmol), and compound 9-2 (4.13 g, 24.73 mmol) were added. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 17-2 (1.66 g, 89% yield).

[0220] Step 2: A mixture of compound 17-2 (1 g, 2.67 mmol) in 4N HCl (EtOAc solution, 10 mL) and MeOH (0.5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to give title compound 17-3 (1 g, crude), which was used in the next step without further purification.

[0221] Step 3: A solution of compound 17-3 (200 mg, 0.673 mmol), compound 8-1 (660 mg, 3.20 mmol), DIPEA (862 mg, 6.68 mmol), and HATU (1.52 g, 3.99 mmol) in DMF (5 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL x 2) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 17-4 (623 mg, 50% yield for two steps). LCMS: C 26 H 42 Calculated m / z for N2O5: 462.63, Found: 463.95 [M+H] + .

[0222] Step 4: To a solution of compound 17-4 (620 mg, 1.34 mmol) in EtOH (6 mL) and HO (3 mL) was added NaOH (214 mg, 5.36 mmol) at 0° C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1N HCl at 0° C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 55% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 17-5 (209 mg, 35% yield) as a white solid. LCMS: C 22 H 34 Calculated m / z for N2O5: 406.52, Found: 407.83 [M+H] + .

[0223] Step 5: A mixture of compound 17-5 (172 mg, 0.388 mmol) and compound 9-7 (149 mg, 0.388 mmol) in toluene (3 mL) was stirred at 120° C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 17 (55 mg, 18% yield) as a white solid. LCMS: C 41 H 55 Calculated m / z for BN6O7: 754.74, Found: 755.93 [M+H] + .

[0224] Example 18. N-((S)-1-(((R)-1-(6-(6-((S)-2-(4-isobutylphenyl)propanamido)hexyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (18)

[0225] [ka]

[0226] [ka]

[0227] Step 1: To a solution of compound 18-1 (1.0 g, 4.62 mmol) in DMF (10 mL), NaHCO3 (3.88 g, 46.18 mmol), KI (767 mg, 4.59 mmol), and compound 9-2 (3.86 g, 23.11 mmol) were added. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 18-2 (1.24 g, 69% yield).

[0228] Step 2: A mixture of compound 18-2 (700 mg, 1.80 mmol) in 4N HCl (in EtOAc, 7 mL) and MeOH (0.5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to give title compound 18-3 (622 mg, crude), which was used in the next step without further purification.

[0229] Step 3: A solution of compound 18-3 (622 mg, crude), compound 8-1 (371 mg, 1.79 mmol), DIPEA (581 mg, 4.50 mmol), and HATU (1.03 g, 2.70 mmol) in DMF (5 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL x 2) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 18-4 (403 mg, 46% yield for two steps). LCMS: C 27 H 44 Calculated m / z for N2O5: 476.66, Found: 478.01 [M+H] + .

[0230] Step 4: To a solution of compound 18-4 (403 mg, 0.845 mmol) in EtOH (4 mL) and HO (2 mL) was added NaOH (135 mg, 3.37 mmol) at 0° C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1N HCl at 0° C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 18-5 (181 mg, 46% yield) as a white solid. LCMS: C 23 H 36 Calculated m / z for N2O5: 420.55, Found: 421.76 [M+H] + .

[0231] Step 5: A mixture of compound 18-5 (178 mg, 0.389 mmol) and compound 9-7 (149 mg, 0.388 mmol) in toluene (3 mL) was stirred at 120° C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 18 (65 mg, 21% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ9.07(d,J=1.4Hz,1H),8.90(d,J=8.4Hz,1H),8.86(d,J=2.5Hz,1H),8.72(dd,J=2.5,1.5Hz,1H),7.87(t,J=5.5Hz ,1H),7.32(d,J=9.8Hz,1H),7.29-7.22(m,4H),7.21-7.13(m,3H),7.05(d,J=8.0Hz,2H),4.67(td,J=8.4,6.4Hz,1H),4.16(d,J=17.4Hz,1 H),4.04(d,J=17.4Hz,1H),3.94(d,J=16.7Hz,1H),3.60-3.50(m,2H),3.46(d,J=16.7Hz,1H),3.09-2.95(m,6H),2.38(d,J=7.1Hz,2H),1 .77(dt,J=13.5,6.7Hz,1H),1.61-1.51(m,2H),1.42-1.33(m,3H),1.29(d,J=7.1Hz,4H),1.26-1.15(m,5H),0.83(td,J=8.3,6.4Hz,12H). LCMS:C 42 H 57 Calculated m / z for BN6O7: 768.76, Found: 769.93 [M+H] + .

[0232] Example 19. N-((S)-1-(((R)-1-(6-(7-((S)-2-(4-isobutylphenyl)propanamido)heptyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (19)

[0233] [ka]

[0234] [ka]

[0235] Step 1: A mixture of compound 8-1 (1 g, 4.84 mmol), NHS (7.27 mmol), DMAP (60 mg, 0.491 mmol), and EDCI (1.86 g, 9.73 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with DCM (50 mL) and washed with 1N HCl (50 mL) and water (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 19-1 (1.26 g, 85% yield) as a white solid.

[0236] Step 2: To a solution of compound 19-2 (996 mg, 7.66 mmol) and DIPEA (987 mg, 7.65 mmol) in DCM (5 mL) was added a solution of compound 19-1 (1.16 g, 3.82 mmol) in DCM (6 mL) over 1 h. The reaction was stirred at room temperature for 4 h, and then the mixture was concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with 80% MeOH in water, HCl condition). Concentration of the desired components afforded the title compound 19-3 (HCl salt, 667 mg, 49% yield) as a colorless oil. LCMS: C 20 H 34 Calculated m / z for NO: 318.51, Measured: 319.88 [M+H] + .

[0237] Step 3: To a solution of compound 19-3 (665 mg, 2.08 mmol) in DMF (8 mL), NaHCO3 (1.75 g, 20.83 mmol), KI (346 mg, 2.08 mmol), and compound 9-2 (1.74 g, 10.41 mmol) were added. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to give the title compound 19-4 (629 mg, 61% yield). LCMS: C 28 H 46 Calculated m / z for N2O5: 490.69, Found: 491.94 [M+H] + .

[0238] Step 4: To a solution of compound 19-4 (627 mg, 1.27 mmol) in EtOH (6 mL) and HO (3 mL) was added NaOH (204 mg, 5.1 mmol) at 0° C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1N HCl at 0° C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 55% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 19-5 (229 mg, 38% yield) as a white solid. LCMS: C 24 H 38 Calculated m / z for N2O5: 434.58, Found: 435.75 [M+H] + .

[0239] Step 5: A mixture of compound 9-5 (195 mg, 0.414 mmol) and compound 9-7 (159 mg, 0.414 mmol) in toluene (5 mL) was stirred at 120° C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 19 (170 mg, 52% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ9.07(d,J=1.5Hz,1H),8.90(d,J=8.4Hz,1H),8.86(d,J=2.5Hz,1H),8.72(q,J=1.8Hz,1H),7.86(t,J=5.6Hz,1H) ,7.35-7.25(m,4H),7.24-7.20(m,2H),7.17(d,J=10.7Hz,2H),7.05(dd,J=8.2,2.0Hz,2H),4.68(td,J=8.3,5.6Hz,1H),4.17(d,J=17.4 Hz,1H),4.04(d,J=17.4Hz,1H),3.95(d,J=16.6Hz,1H),3.60-3.50(m,2H),3.47(d,J=17.0Hz,1H),3.10-2.97(m,6H),2.37(dd,J=7.2,2 .5Hz,2H),1.78(ddd,J=13.2,6.6,2.2Hz,1H),1.56(dp,J=10.3,3.4Hz,2H),1.42-1.33(m,3H),1.31-1.11(m,11H),0.86-0.80(m,12H). LCMS:C 43 H 59 Calculated m / z for BN6O7: 782.79, Found: 783.86 [M+H] + .

[0240] Example 20. N-((S)-1-(((R)-1-(6-(8-((S)-2-(4-isobutylphenyl)propanamido)octyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (20)

[0241] [ka]

[0242] [ka]

[0243] Step 1: To a solution of compound 20-1 (1.0 g, 4.09 mmol) in DMF (10 mL), NaHCO3 (3.44 g, 40.94 mmol), KI (679 mg, 4.09 mmol), and compound 9-2 (3.42 g, 20.47 mmol) were added. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 20-2 (1.25 g, 69% yield).

[0244] Step 2: A mixture of compound 20-2 (700 mg, 1.68 mmol) in 4N HCl (EtOAc solution, 7 mL) and MeOH (0.5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to give title compound 20-3 (637 mg, crude), which was used in the next step without further purification.

[0245] Step 3: A solution of compound 20-3 (637 mg, crude from step 2), compound 8-1 (346 mg, 1.67 mmol), DIPEA (542 mg, 4.20 mmol), and HATU (958 mg, 2.52 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL×2) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 20-4 (362 mg, 42% yield for two steps). LCMS: C 29 H 48 Calculated m / z for N2O5: 504.71, Found: 506.07 [M+H] + .

[0246] Step 4: To a solution of compound 20-4 (360 mg, 0.713 mmol) in EtOH (4 mL) and HO (2 mL) was added NaOH (114 mg, 2.85 mmol) at 0° C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1N HCl at 0° C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 20-5 (247 mg, 71% yield) as a white solid. LCMS: C 25 H 40 Calculated m / z for N2O5: 448.60, Found: 449.95 [M+H] + .

[0247] Step 5: A mixture of compound 20-5 (178 mg, 0.367 mmol) and compound 9-7 (141 mg, 0.366 mmol) in toluene (3 mL) was stirred at 120° C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 20 (50 mg, 17% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ9.07(d,J=1.5Hz,1H),8.90(d,J=8.5Hz,1H),8.86(d,J=2.5Hz,1H),8.73(t,J=1.9Hz,1H),7.85(t,J=5.6Hz,1H),7 .32(d,J=9.7Hz,1H),7.29-7.23(m,4H),7.23-7.17(m,3H),7.05(d,J=7.8Hz,2H),4.68(td,J=8.7,5.8Hz,1H),4.17(d,J=17.4Hz,1H),4.0 4(d,J=17.4Hz,1H),3.95(d,J=16.6Hz,1H),3.60-3.51(m,2H),3.48(d,J=16.6Hz,1H),3.08-2.97(m,6H),2.38(d,J=7.1Hz,2H),1.78(dt, J=13.5,6.7Hz,1H),1.61-1.52(m,2H),1.37(dd,J=21.4,4.9Hz,3H),1.28(d,J=7.1Hz,4H),1.26-1.17(m,9H),0.83(q,J=8.5,7.5Hz,12H). LCMS:C 44 H 61 Calculated m / z for BN6O7: 796.82, Found: 797.99 [M+H] + .

[0248] Example 21. N-((S)-1-(((R)-1-(6-(4-(((S)-2-(4-isobutylphenyl)propanamido)methyl)cyclohexyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (21)

[0249] [ka]

[0250] [ka]

[0251] Step 1: A mixture of compound 21-1 (700 mg, 3.07 mmol), 10 w% Pd / C (350 mg), and ammonium hydroxide (28%, 9.5 mL) in MeOH (10 mL) was purged with hydrogen three times and stirred under hydrogen atmosphere at room temperature for 16 h. The mixture was concentrated to give the title compound 21-2 (746 mg, crude) as a colorless oil, which was used in the next step without further purification.

[0252] Step 2: To a solution of compound 21-2 (380 mg, 1.66 mmol) in DMF (5 mL), NaHCO3 (1.39 g, 16.54 mmol), KI (275 mg, 1.65 mmol), and compound 9-2 (1.38 g, 8.26 mmol) were added. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to give the title compound 21-3 (490 mg, 73% yield for two steps).

[0253] Step 3: A mixture of compound 21-3 (480 mg, 1.19 mmol) in 4N HCl (EtOAc solution, 5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to give title compound 21-4 (450 mg, crude), which was used in the next step without further purification.

[0254] Step 4: A solution of compound 21-4 (450 mg), compound 8-1 (297 mg, 1.43 mmol), DIPEA (390 mg, 3.02 mmol), and HATU (684 mg, 1.79 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL x 2) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to give the title compound 21-5 (339 mg, 58% yield). 1H NMR (400MHz, chloroform-d) δ7.17(d,J=7.1Hz,2H),7.11(d,J=7.6Hz,2H),5.30(s,1H),4.14( q,J=7.1Hz,4H),3.52(s,4H),3.49(d,J=7.9Hz,1H),2.99(q,J=6.0Hz,2H),2.57(t,J=11. 6Hz,1H),2.45(d,J=7.0Hz,2H),1.83(d,J=8.2Hz,3H),1.76(s,1H),1.60(d,J=12.7Hz,2H ),1.50(d,J=7.1Hz,3H),1.25(t,J=7.1Hz,8H),1.11(q,J=12.0Hz,2H),0.91-0.87(m,6H). LCMS:C 28 H 44 Calculated m / z for N2O5: 488.67, Measured: 489.75 [M+H] + .

[0255] Step 5: To a solution of compound 21-5 (337 mg, 0.689 mmol) in EtOH (3 mL) and HO (1.5 mL) was added NaOH (110 mg, 2.75 mmol) at 0° C. The reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1N HCl at 0° C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 55% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 21-6 (207 mg, 64% yield) as a white solid.

[0256] Step 6: A mixture of compound 21-6 (155 mg, 0.33 mmol) and compound 9-7 (127 mg, 0.33 mmol) in toluene (3 mL) was stirred at 120° C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 21 (115 mg, 44% yield) as an off-white solid. 1H NMR(400MHz,DMSO-d6)δ9.08(d,J=1.4Hz,1H),8.89-8.85(m,2H),8.73(dd,J=2. 5,1.5Hz,1H),7.92(t,J=5.9Hz,1H),7.46(d,J=9.5Hz,1H),7.32-7.28(m,2H),7. 25(t,J=7.5Hz,2H),7.21(d,J=8.0Hz,2H),7.19-7.15(m,1H),7.05(d,J=8.0Hz, 2H),4.72-4.64(m,1H),4.31(d,J=17.9Hz,1H),3.84(dd,J=17.0,6.7Hz,2H),3.6 2-3.52(m,2H),3.25(d,J=16.4Hz,2H),3.07(d,J=7.6Hz,2H),2.86(t,J=6.5Hz, 2H),2.39(d,J=7.1Hz,2H),1.77(dt,J=13.7,6.8Hz,1H),1.74-1.62(m,3H),1.61 -1.36(m,6H),1.31(d,J=7.1Hz,4H),1.16(ddd,J=13.7,10.4,2.9Hz,1H),1.09- 0.97(m,1H),0.86(d,J=6.7Hz,3H),0.83(d,J=6.6Hz,6H),0.78(d,J=6.4Hz,3H). LCMS:C 43 H 57 Calculated m / z for BN6O7: 780.77, Found: 782.00 [M+H] + .

[0257] Example 22. N-((S)-1-(((R)-3-methyl-1-(6-(4-((S)-2-(4-(2-(methyl-d3)propyl-3,3,3-d3)phenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)butyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (22)

[0258] [ka]

[0259] [ka]

[0260] Step 1: NaH (60% in mineral oil, 9.35 g, 233 mmol) was added portionwise over 10 min to a solution of compound 22-2 (63 g, 281 mmol) in anhydrous THF (250 mL) at 0° C. under nitrogen atmosphere. After stirring at 0° C. for 1 h, deuterated acetone 22-1 (10 g, 156 mmol) was added slowly over 15 min. The reaction was then warmed to room temperature and stirred for an additional 6 h. The mixture was quenched with saturated NH4Cl solution (300 mL) and extracted with isopropyl ether (300 mL×2). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 22-3 (22 g, crude), which was used in the next step without further purification.

[0261] Step 2: 2N LiOH (270 mL, 540 mmol) was added to a mixture of compound 22-3 (22 g, crude from step 1) in EtOH (150 mL) at 0° C. The reaction was stirred at room temperature for 16 h. The resulting mixture was cooled to 0° C. and adjusted to pH=2 with 6N aqueous HCl. The mixture was extracted with isopropyl ether (200 mL×2). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 22-4 (13.4 g, 80% for two steps), which was used in the next step without further purification.

[0262] Step 3: To a solution of compound 22-4 (3 g, 28.2 mmol), compound 22-5 (6.92 g, 42.4 mmol), and DMAP (170 mg, 1.39 mmol) in DCM (50 mL) was added DCC (8.75 g, 42.4 mmol) at 0° C. The reaction was stirred at room temperature for 15 h. The mixture was concentrated and purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to give the title compound 22-6 (3.2 g, 45% yield) as a white solid.

[0263] Step 4: A mixture of compound 22-6 (2.3 g, 9.15 mmol), compound 22-7 (330 mg, 1.84 mmol), and B2(Pin)2 (4.65 g, 18.3 mmol) in anhydrous PhCF3 (23 mL) was purged with nitrogen three times and stirred at 110° C. for 20 h. The mixture was concentrated and purified by silica gel column (eluted with 3% EtOAc in petroleum ether) to give the title compound 22-8 (470 mg, 27% yield) as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δ4.98 (s, 1H), 1.19 (s, 12H).

[0264] Step 5: A mixture of compound 22-8 (550 mg, 2.92 mmol), compound 22-9 (736 mg, 3.21 mmol), Pd(PPh3)4 (168 mg, 0.145 mmol), and K2CO3 (1.2 g, 8.68 mmol) in dioxane (6 mL) under nitrogen was stirred at 120 °C for 16 h. After completion of the reaction, the mixture was filtered and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, TFA condition). The desired component was lyophilized to give the title compound 22-10 (460 mg, 75% yield) as a colorless slurry.

[0265] Step 6: A mixture of compound 22-10 (460 mg, 2.18 mmol) and PtO2 (50 mg, 0.22 mmol) in MeOH (5 mL) was stirred under hydrogen atmosphere for 16 h. After completion of the reaction, the mixture was filtered over celite and concentrated to give the title compound 22-11 (350 mg, 75% yield) as a colorless slurry. 1 H NMR(400MHz,DMSO-d6)δ7.18(d,J=7.8Hz,2H),7.08(d,J=7.8Hz,2H),3.60(q,J= 7.1Hz,1H), 2.40(d,J=7.2Hz,2H),1.76(t,J=7.2Hz,1H),1.32(d,J=7.1Hz,3H).

[0266] Step 7: A solution of compound 13-2 (200 mg, 0.673 mmol), compound 22-11 (95 mg, 0.447 mmol), DIPEA (260 mg, 2.01 mmol), and HATU (300 mg, 0.789 mmol) in DMF (3 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with EtOAc (30 mL) and washed with water (30 mL×2) and brine (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to give the title compound 22-12 (106 mg, 52% yield). LCMS: C 25 H 34 Calculated m / z for D6N2O5: 454.64, Found: 455.99 [M+H] + .

[0267] Step 8: To a solution of compound 22-12 (100 mg, 0.220 mmol) in EtOH (and 2 mL) was added 2N NaOH (0.44 mL, 0.88 mmol) at 0° C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1N HCl at 0° C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 35% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 22-13 (68 mg, 75% yield) as a white solid.

[0268] Step 9: A mixture of compound 22-13 (60 mg, 0.151 mmol) and compound 9-7 (60 mg, 0.156 mmol) in toluene (3 mL) was stirred at 120° C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 22 (26 mg, 23% yield) as an off-white solid. 1H NMR(400MHz,DMSO-d6)δ9.12(s,1H),8.90(s,2H),8.76(s,1H),8.00(s,1H),7.38(d,J=9.7Hz,1H),7.27(ddt,J= 24.5,17.1,7.2Hz,7H),7.09(d,J=7.7Hz,2H),4.72(d,J=7.9Hz,1H),4.10(q,J=17.3Hz,2H),3.95(d,J=16.6Hz, 1H),3.58(d,J=8.2Hz,2H),3.50(d,J=17.1Hz,1H),3.17-3.01(m,6H),2.41(d,J=7.0Hz,2H),1.78(d,J=8.4Hz,1 H),1.66-1.54(m,2H),1.43(d,J=12.5Hz,4H),1.35(d,J=6.8Hz,3H),1.20(s,1H),0.87(dd,J=16.0,6.7Hz,6H). LCMS:C 40 H 47 Calculated m / z for D6BN6O7: 746.75, Found: 747.5 [M+H] + .

[0269] Example 23. N-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-4-(4-iodophenyl)butanamide (23)

[0270] [ka]

[0271] [ka]

[0272] Step 1: To an ice-cold solution of compound 23-1 (10.0 g, 52.8 mmol) in anhydrous THF (200 mL) was added PPh3 (20.8 g, 79.3 mmol) followed by a solution of CBr4 (26.3 g, 79.3 mmol) in THF (100 mL). The reaction was allowed to warm to room temperature and stirred for 2 h. The mixture was filtered and concentrated. The residue was purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to give the title compound 23-2 (12 g, 90%) as a colorless oil.

[0273] Step 2: A mixture of compound 23-3 (500 mg, 1.95 mmol), K2CO3 (540 mg, 3.90 mmol), and compound 23-2 (590 mg, 2.34 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. The resulting mixture was filtered and the cake was washed with EtOAc (50 mL). The organic phase was washed with cold water (50 mL) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 5% MeOH in dichloromethane) to give the title compound 23-4 (585 mg, 70% yield) as a pale yellow solid.

[0274] Step 3: 4N HCl (in EtOAc, 2.5 mL) was added to a solution of compound 23-4 (585 mg, 1.36 mmol) in EtOAc (2 mL) and MeOH (0.5 mL). The reaction was stirred at room temperature for 1 h, and the mixture was concentrated to give the title compound 23-5 (450 mg, 90% yield) as a white solid.

[0275] Step 4: A mixture of compound 23-5 (80 mg, 0.275 mmol), DIPEA (56 mg, 0.434 mmol), DMAP (3 mg, 0.024 mmol), EDCI (65 mg, 0.340 mmol), and compound 1-2 (80 mg, 0.220 mmol) in DCM (5 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-HPLC (eluted with 60% acetonitrile in water, HCl condition) to give 23 (60 mg, 45% yield) as a white solid.1 H NMR(500MHz,DMSO-d6)δ8.19(d,J=8.1Hz,1H),7.90(d,J=5.4Hz,1H),7.84(d,J=8.4Hz,1H),7.77-7 .72(m,1H),7.64-7.56(m,3H),7.00(d,J=8.1Hz,2H),4.41(t,J=6.6Hz,2H),3.15(q,J=6.5Hz,2H),3 .00(t,J=7.6Hz,2H),2.52(d,J=7.9Hz,2H),2.07(t,J=7.4Hz,2H),1.92(t,J=7.5Hz,2H),1.82(t,J =7.6Hz,2H),1.77(t,J=7.5Hz,2H),1.71-1.62(m,2H),1.42(q,J=7.4Hz,2H),0.93(t,J=7.4Hz,3H). LCMS:C 28 H 34 Calculated m / z for IN5O2: 599.52, Found: 600.82 [M+H] + .

[0276] Example 24. N-(4-((4-amino-2-butyl-7-(dimethylphosphoryl)-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-4-(4-iodophenyl)butanamide (24)

[0277] [ka]

[0278] [ka]

[0279] Step 1: A mixture of compound 24-1 (19 mg, 0.048 mmol), compound 1-2 (28 mg, 0.0965 mmol), EDCI (36 mg, 0.188 mmol), DMAP (1 mg, 0.008 mmol), and TEA (10 mg, 0.099) in DCM (0.5 mL) and DMF (0.5 mL) was stirred at room temperature for 1 h. After completion of the reaction, the resulting mixture was filtered and purified by pre-HPLC (eluted with 70% acetonitrile in water, HCl condition). The components were lyophilized to give 24 (11.2 mg, 35.8% yield) as a yellow oil. LCMS: C 30 H 39 Calculated m / z for IN5O3P: 675.55, Found: 676.2 [M+H] + .

[0280] Example 25. N-(2-((2-((4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl(3-(dimethylamino)propyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)ethyl)-4-(4-iodophenyl)butanamide (25)

[0281] [ka]

[0282] [ka]

[0283] Step 1: To a solution of compound 1-2 (400 mg, 1.37 mmol) in DCM (8 mL) was added EDCI (395 mg, 2.06 mmol) and compound 9-1 (243 mg, 1.51 mmol). After stirring at room temperature for 2 h, TLC showed that compound 1-2 was consumed. The reaction mixture was diluted with EtOAc and washed with 1N HCl, saturated Na2CO3 solution, and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the title compound 25-1 (330 mg) as a colorless oil, which was used directly in the next step. TLC (petroleum ether: EtOAc / 1:1):R f (Compound 9-1)=0.55, R f (Compound 25-1)=0.37.

[0284] Step 2: A mixture of crude compound 25-1 (330 mg) in HCl / EtOAc (4M, 10 mL) was stirred at room temperature for 30 min. After completion of the reaction, the mixture was concentrated. The residue was redissolved in EtOAc and washed with saturated Na2CO3 solution. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the title compound 25-2 (225 mg) as a colorless oil, which was used directly in the next step. TLC (petroleum ether: EtOAc / 1:1) R f (Compound 25-1)=0.37, TLC(DCM:MeOH / 10:1):R f (Compound 25-2)=0.46.

[0285] Step 3: To a solution of compound 25-3 (154 mg, 0.905 mmol) in MeOH (5 mL) was added a solution of compound 25-2 (225 mg, crude) in MeOH (2 mL) dropwise at 0° C. After stirring at room temperature for 4 h, TLC showed that compound 25-2 was consumed and a new spot with lower polarity was formed. The reaction mixture was evaporated under vacuum and purified by silica gel column (eluted with 5% MeOH in DCM) to give the title compound 25-4 (230 mg, 36% yield for three steps) as a white solid. TLC (DCM:MeOH / 10:1):R f (Compound 25-2)=0.46, R f (Compound 25-4)=0.58.

[0286] Step 4: A mixture of compound 25-4 (67 mg, 0.145 mmol), compound 25-5 (40 mg, 0.970 mmol), and DIPEA (40 mg, 0.396 mmol) in acetonitrile (2 mL) and DCM (2 mL) was stirred at room temperature for 24 h. LC-MS showed that compound 25-5 was consumed and one new peak with the desired m / z was detected by LCMS. The reaction mixture was concentrated, redissolved in acetonitrile and water, and purified by HPLC (C18 column, eluted with acetonitrile / H2O, HCl condition). The desired component was lyophilized to give 25 (8 mg, 10% yield) as a white powder. 1 HNMR (400MHz, methanol-d4) δ8.12(d,J=8.1Hz,1H),7.68(d,J=8.4Hz,1H),7.53(t,J=7.8Hz,1H),7.46(d,J=7.9Hz,2 H),7.36(t,J=7.7Hz,1H),6.86(d,J=7.9Hz,2H),4.33(s,2H),3.75(t,J=5.8Hz,2H),3.41(s,2H),3.35(d,J=2.4H) z,2H),2.99(t,J=7.6Hz,2H),2.47(t,J=7.8Hz,2H),2.39(d,J=7.1Hz,2H),2.27(s,6H),2.16(d,J=7.7Hz,2H),1. 98(d,J=9.8Hz,2H),1.85(dq,J=23.9,7.5Hz,8H),1.49(d,J=7.6Hz,2H),1.30-1.26(m,2H),1.01(t,J=7.4Hz,3H). LCMS:C 39 H 51 Calculated m / z for IN8O4: 822.79, Found: 823.4 [M+H] + .

[0287] Example 26. (S)-N-(2-((2-((4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)ethyl)-2-(4-isobutylphenyl)propanamide (26)

[0288] [ka]

[0289] [ka]

[0290] Step 1: To a solution of compound 25-3 (1.06 g, 6.23 mmol) in anhydrous MeOH (10 mL) was added a solution of compound 9-1 (1 g, 6.24 mmol) in anhydrous MeOH (10 mL) dropwise. The mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by column (eluted with 40% EtOAc in petroleum ether) to give the title compound 26-1 (1.63 g, 97% yield) as a colorless oil.

[0291] Step 2: To a solution of compound 23-5 (339 mg, 1.03 mmol) and DIPEA (333 mg 2.58 mmol) in anhydrous MeOH (4 mL) was added a solution of compound 26-1 (440 mg, 1.55 mmol) in anhydrous MeOH (3 mL) dropwise. The mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with 60% acetonitrile in water, TFA condition). The desired component was lyophilized to give the title compound 26-2 (TFA salt, 445 mg, 63% yield) as a yellow solid.

[0292] Step 3: A mixture of compound 26-2 (445 mg, 0.65 mmol) in a solution of 20% TFA in DCM (3 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated to remove the organic solvent to give the title compound 26-3 (420 mg, 92% yield) as a pale yellow slurry.

[0293] Step 4: A mixture of compound 26-3 (100 mg, 0.148 mmol), compound 8-1 (46 mg, 0.222 mmol), DMAP (2 mg, 0.016 mmol), DIPEA (57 mg, 0.442 mmol), and EDCI (42 mg, 0.220 mmol) in DMF (2 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was filtered and purified by pre-HPLC (eluted with 50% acetonitrile in water, HCl condition). The desired component was lyophilized to give 26 (40 mg, 39% yield) as a white solid. 1 H NMR(600MHz,DMSO-d6)δ8.20(dd,J=8.1,1.4Hz,1H),7.85-7.81(m,1H),7.74(ddd,J=8.4,7.2,1.3Hz,1H),7.62(t,J=7 .7Hz,1H),7.17(d,J=7.5Hz,2H),7.01(d,J=7.2Hz,2H),4.44(t,J=6.5Hz,2H),3.57-3.43(m,5H),3.28(d,J=7.0Hz,1H ),3.12(dd,J=13.3,6.5Hz,1H),3.00(t,J=7.6Hz,2H),2.39-2.31(m,2H),2.01(s,2H),1.82(td,J=14.1,13.2,6.6Hz, 4H), 1.76(d,J=4.4Hz,1H),1.43(q,J=7.4Hz,2H),1.29(d,J=7.0Hz,3H),0.93(t,J=7.4Hz,3H),0.81(d,J=6.6Hz,6H). LCMS:C 37 H 47 Calculated m / z for N7O4: 653.83, Found: 654.88 [M+H] + .

[0294] Example 27. (S)-N-(4-((4-amino-2-butyl-7-(dimethylphosphoryl)-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-2-(4-isobutylphenyl)propanamide (27)

[0295] [ka]

[0296] [ka]

[0297] Step 1: A mixture of compound 24-1 (60 mg, 0.148 mmol), compound 8-1 (46 mg, 0.223 mmol), DMAP (2 mg, 0.016 mmol), DIPEA (57 mg, 0.442 mmol), and EDCI (45 mg, 0.235 mmol) in DMF (2 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was filtered and purified by pre-HPLC (eluted with 45% acetonitrile in water, HCl condition). The desired component was lyophilized to give 27 (24 mg, 27% yield) as a white solid. 1 H NMR(600MHz,DMSO-d6)δ8.29-8.24(m,2H),8.09(t,J=5.7Hz,1H),7.95(ddd,J=10.0,8.3,1.3Hz, 1H),7.23(d,J=8.1Hz,2H),7.05(d,J=8.1Hz,2H),4.39(t,J=6.6Hz,2H),3.60(s,1H),3.16(q,J= 6.6Hz,2H),3.04-3.00(m,2H),2.37(d,J=7.1Hz,2H),1.91-1.82(m,4H),1.77(d,J=13.5Hz,7H), 1.68-1.63(m,2H),1.49-1.41(m,2H),1.32(d,J=7.1Hz,3H),0.96(t,J=7.4Hz,3H),0.82(s,6H). LCMS:C 33 H 46 Calculated m / z for N5O3P: 591.74, Found: 592.93 [M+H] + .

[0298] Example 28 (5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methyl 4-(4-iodophenyl)butanoate (28)

[0299] [ka]

[0300] [ka]

[0301] Step 1: Compound 28-1 (1.3 g, 10.0 mmol) was dissolved in 37% aqueous formaldehyde (1.8 g, 22.2 mmol), and the mixture was refluxed with stirring for 4 h at 60° C. The mixture was concentrated to give the title compound 28-2 (1.5 g, 93% yield) as a colorless oil, which was used directly in the next step without purification.

[0302] Step 2: Oxalyl chloride (0.50 mL, 5.82 mmol) and catalytic amount of DMF were added to a solution of compound 1-2 (200 mg, 0.69 mmol) in dichloromethane (20 mL). The resulting mixture was stirred at 45° C. for 1 h. The mixture was azeotroped with dichloromethane 3 under reduced pressure to give the crude chloride intermediate. The chloride was redissolved in anhydrous dichloromethane (5 mL) and added to a solution of compound 28-2 (220 mg, 1.38 mmol), DMAP (8 mg, 0.07 mmol), and TEA (100 uL, 0.69 mmol) in anhydrous dichloromethane (10 mL) at room temperature. The resulting mixture was refluxed at 50° C. with stirring for 10 h. After the mixture was concentrated, the residue was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to give 28 (72 mg, 26% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ12.00(s,1H),8.14(d,J=6.4Hz,1H),7.68-7.56(m,2H),7.01(d,J=7 .8Hz,2H),5.56(s,2H),2.54(d,J=7.6Hz,2H),2.33(t,J=7.3Hz,2H),1.78(t,J=7.5Hz,2H).

[0303] Example 29. (5-Fluoro-2,4-dioxopyrimidine-1,3(2H,4H)-diyl)bis(methylene)bis(4-(4-iodo-phenyl)butanoate (29)

[0304] [ka]

[0305] [ka]

[0306] Step 1: A suspension of compound 28-1 (1.0 g, 7.69 mmol) in 37% formaldehyde solution (2 mL) was stirred for 4 h at 60° C. The reaction mixture was concentrated to give the title compound 29-1 (1.5 g) as a colorless oil, which was used directly in the next step without purification.

[0307] Step 2: To a solution of compound 1-2 (500 mg, 1.73 mmol) in DCM (5 mL), oxalyl chloride (2.18 g, 17.21 mmol) and catalytic amount of DMF were added at room temperature. The resulting mixture was stirred at 45° C. for 1 h. After compound 1-2 was consumed, the mixture was concentrated. The residue was redissolved in DCM (10 mL), and to this solution, compound 29-1 (250 mg, 1.32 mmol), DMAP (16 mg, 0.131 mmol), and TEA (650 mg, 6.44 mmol) were added at room temperature. The resulting mixture was stirred at 50° C. for 16 h. After completion of the reaction, the mixture was concentrated. The residue was purified by silica gel column (eluted with 2% EtOAc in petroleum ether) and further purified by reverse-phase flash chromatography (C18 column, eluted with acetonitrile and water, TFA condition) to give 29 (70 mg, 7% yield for two steps) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ8.29(dq,J=6.3,3.8,3.1Hz,1H),7.61(ddq,J=6.4,4.1,2.4Hz,4H),6.99(dt,J=8.1,3.4Hz ,4H),5.82-5.75(m,2H),5.67-5.58(m,2H),2.52(d,J=5.9Hz,4H),2.35-2.23(m,4H),1.76(dq,J=12.2,7.2Hz,4H). LCMS:C 26 H 25m / z calculated for FI2N2O6: 734.30, found: 757.33. [M+Na] + .

[0308] Example 30. (2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-12b-acetoxy-9-(((2R,3S)-3-((tert-butoxycarbonyl)amino)-2-((4-(4-iodophenyl)butanoyl)oxy)-3-phenylpropanoyl)oxy)-4,6,11-trihydroxy-4a,8,13,13-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-1H-7,11-methanocyclodeca[3,4]benzo[1,2-b]oxet-12-yl benzoate (30)

[0309] [ka]

[0310] [ka]

[0311] Step 1: DCC (77 mg, 0.37 mmol) and DMAP (45 mg, 0.37 mmol) were added to a solution of compound 30-1 (200 mg, 0.25 mmol) and 4-(p-iodophenyl)butyric acid 1-2 (79 mg, 0.27 mmol) in dichloromethane (16 mL) at -10 °C. After stirring at -10 °C for 3 h, the mixture was filtered, diluted with EtOAc (50 mL), and washed with water (50 mL) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel column (eluted with 40% EtOAc in petroleum ether) to give 30 (115 mg, yield: 43%) as a white solid. 1H NMR (400MHz, chloroform-d) δ8.16(d,J=7.7Hz,2H),7.68-7.60(m,3H),7.55(t,J=7.5Hz,2H ),7.43(t,J=7.5Hz,2H),7.37-7.31(m,3H),6.90(d,J=7.8Hz,2H),6.30(s,1H),5.74(d ,J=7.0Hz,1H),5.52(s,1H),5.45-5.34(m,2H),5.26(s,1H),5.01(d,J=9.4Hz,1H),4.3 7(d,J=8.5Hz,1H),4.31(q,J=8.3Hz,1H),4.24(d,J=6.4Hz,2H),4.20-4.13(m,1H),3.9 9(d,J=7.0Hz,1H),2.64(dt,J=15.4,8.1Hz,1H),2.54(t,J=7.6Hz,3H),2.49(s,3H),2. 38(dp,J=23.3,8.3Hz,4H),2.22(s,1H),2.09(d,J=1.7Hz,2H),2.01(s,3H),1.90(q,J= 8.4,7.7Hz,4H),1.80(s,3H),1.69(s,1H),1.63(d,J=1.7Hz,3H),1.54(d,J=7.5Hz,1H) ,1.38(s,11H),1.32(d,J=1.9Hz,1H),1.30(d,J=1.7Hz,2H),1.28(s,4H),1.17(s,4H). LCMS:C 53 H 62 INO 15 Calculated m / z: 1079.98, Measured: 1102.7 [M+Na] + .

[0312] Example 31.N 2 -(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl)benzoyl)-N 5 -(2-(4-(4-iodophenyl)butanamido)ethyl)-L-glutamine (31)

[0313] [ka]

[0314] [ka]

[0315] Step 1: EDCI (362 mg, 1.89 mmol), triethylamine (0.74 mL, 5.16 mmol), and DMAP (20 mg, 0.17 mmol) were added to a solution of compound 31-1 (520 mg, 1.41 mmol) and compound 25-2 (574 mg, 1.89 mmol) in dichloromethane (20 mL) at 0° C. The reaction was stirred at room temperature for 2 h, and the mixture was diluted with dichloromethane (100 mL) and washed with 1N HCl (100 mL) and brine (100 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 31-2 (740 mg, yield: 85%), which was used in the next step without further purification.

[0316] Step 2: A mixture of compound 31-2 (400 mg, 0.65 mmol) in 4N HCl (EtOAc solution, 5 mL) was stirred at room temperature for 1 h. A white precipitate formed and was filtered. The solid was dried under vacuum to give the title compound 31-3 (HCl salt, 200 mg, 62% yield).

[0317] Step 3: A solution of compound 31-4 (200 mg, 0.67 mmol), DMAP (8 mg, 0.07 mmol), NHS (115 mg, 1.00 mmol), and EDCI (128 mg, 0.67 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 1 h. Compound 31-3 (200 mg, 0.40 mmol) and Et3N (483 uL, 3.35 mmol) were added. After stirring at room temperature for 12 h, the mixture was purified by preparative HPLC (eluted with 60% acetonitrile in water, HCl condition) to give 31 (67 mg, yield: 22%) as a light pink solid. 1H NMR(400MHz,DMSO-d6)δ10.71(s,1H),8.60(d,J=7.7Hz,1H),7.89(s,1H),7.79(d,J=7.7Hz,3H),7.61(d,J=7.9Hz,2H),7.29(d,J=7.9Hz,2H), 7.00(d,J=8.0Hz,2H),6.34(s,1H),6.27(s,1H),4.32(d,J=10.2Hz,1H ),3.12-2.80(m,12H),2.11(dt,J=69.0,7.7Hz,7H),1.79-1.67(m,2H). LCMS:C 32 H 36 m / z calculated for IN7O6: 741.59, found: 742.4. [M+H] + .

[0318] Example 32. (S)-2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl)benzamido)-5-(2-(4-(4-iodophenyl)butanamido)ethoxy)-5-oxopentanoic acid (32)

[0319] [ka]

[0320] [ka]

[0321] Step 1: To a mixture of acid 1-2 (500 mg, 1.72 mmol) and NHS (295 mg, 2.56 mmol) in DCM (10 mL), EDCI (658 mg, 3.44 mmol) and DMAP (21 mg, 0.172 mmol) were added and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with DCM (50 mL) and washed with 1N HCl (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give the crude activated NHS ester as a white solid. To a solution of compound 32-1 (126 mg, 2.06 mmol) and DIEA (340 uL, 2.06 mmol) in DMF (5 mL), a solution of the activated NHS ester in THF (5 mL) was added dropwise. The reaction mixture was stirred at room temperature for 16 h. The resulting reaction mixture was diluted with EtOAc (50 mL) and washed with 1N HCl (50 mL) and brine. The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 32-2 (495 mg, 86% yield) as a white solid, which was used in the next step without further purification.

[0322] Step 2: EDCI (218 mg, 1.14 mmol) was added to a solution of DMAP (7 mg, 0.06 mmol), compound 32-2 (190 mg, 0.57 mmol), and compound 31-1 (260 mg, 0.86 mmol) in DCM (10 mL) at 0° C. After stirring at room temperature for 2 h, the mixture was diluted with DCM (100 mL) and washed with 1N HCl (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4, filtered, and concentrated to give the title compound 32-3 (845 mg, 85% yield), which was used in the next step without further purification.

[0323] Step 3: Compound 32-3 (400 mg, 0.65 mmol) was dissolved in DCM (3 mL) and TFA (3 mL), and the reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated to give the title compound 32-4 (240 mg, 91% yield) as a pale yellow oil.

[0324] Step 4: A mixture of compound 31-4 (200 mg, 0.67 mmol), DMAP (8 mg, 0.07 mmol), NHS (115 mg, 1.00 mmol), and EDCI (128 mg, 0.67 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 1 h. To this mixture was added compound 32-4 (120 mg, 0.26 mmol) and Et3N (0.483 mL, 3.35 mmol). The mixture was stirred at room temperature for 12 h and purified by preparative HPLC (eluted with 60% acetonitrile in water, HCl condition) to give 32 (55 mg, 28% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ8.60(d,J=7.8Hz,1H),8.01-7.90(m,1H),7.79(d,J=7.8Hz,2H),7.67-7.58 (m,2H),7.31(d,J=7.8Hz,2H),7.01(d,J=7.8Hz,2H),6.34(s,1H),4.45(dd,J=9.3,5.5Hz,1H),4.03 (q,J=5.7Hz,2H),3.28(t,J=5.6Hz,2H),3.00(t,J=7.7Hz,2H),2.88(t,J=7.5Hz,2H),2.46(dd,J=1 9.7,7.7Hz,4H),2.22-2.11(m,1H),2.07(t,J=7.4Hz,2H),2.04-1.94(m,1H),1.77(q,J=7.4Hz,2H). LCMS:C 32 H 35 m / z calculated for IN6O7: 742.57, found: 743.3. [M+H] + .

[0325] Example 33. (S)-2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl)benzamido)-5-(4-((S)-2-(4-isobutylphenyl)propanoyl)piperazin-1-yl)-5-oxopentanoic acid (33)

[0326] [ka]

[0327] [ka]

[0328] Step 1: EDCI (3.7 g, 19.37 mmol) was added to a mixture of acid 8-1 (2.00 g, 9.69 mmol), amine 33-1 (2.70 g, 14.49 mmol), and DMAP (118 mg, 0.967 mmol) in DCM (20 mL), and the mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with DCM (50 mL) and washed with 1N HCl (50 mL x 2) and brine (50 mL x 2). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 33-2 (3.62 g, crude) as a white solid, which was used in the next step without further purification. LCMS: C 22 H 34 Calculated m / z for N2O3: 374.53, Found: 397.47. [M+Na] + .

[0329] Step 2: To a solution of compound 33-2 (3.62 g, crude) in EtOAc (30 mL) was added 4N HCl in EtOAc (30 mL). The mixture was stirred at room temperature for 1 h. The mixture was diluted with EtOAc (30 mL) and filtered. The precipitate was washed with EtOAc and dried to give the title compound 33-3 (2.7 g, 89% yield for two steps) as a white solid. LCMS: C 17 H 26 Calculated m / z for NO: 274.41, Found: 275.90. [M+H] + .

[0330] Step 3: EDCI (3.12 g, 16.33 mmol) was added to a solution of DMAP (200 mg, 1.64 mmol), TEA (1.75 g, 17.32 mmol), compound 33-3 (2.70 g, 8.68 mmol), and compound 31-1 (2.48 g, 8.17 mmol) in DCM (40 mL) at 0° C. After stirring at room temperature for 2 h, the mixture was diluted with DCM (100 mL) and washed with 1N HCl (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to give the title compound 33-4 (3.66 g, 80% yield) as a white solid. LCMS: C 31 H 49 m / z calculated for N3O6: 559.75, found: 582.98. [M+Na] + .

[0331] Step 4: Compound 33-4 (2.88 g, 5.00 mmol) was dissolved in DCM (10 mL) and TFA (10 mL) and the reaction was stirred at room temperature for 2 h. The mixture was concentrated to give the title compound 33-5 (2.6 g, 90% yield) as a pale yellow oil. LCMS: 22 H 33 Calculated m / z for N3O4: 403.52, Found: 404.95. [M+H] + .

[0332] Step 5: A mixture of compound 31-4 (300 mg, 1.00 mmol), DMAP (12 mg, 0.098 mmol), NHS (174 mg, 1.51 mmol), and EDCI (383 mg, 2.00 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 2 h. After the activated NHS ester was completely formed, compound 33-5 (560 mg, 1.08 mmol) and Et3N (508 mg, 5.02 mmol) were added. The mixture was stirred at 30° C. for 12 h and purified by preparative HPLC (eluted with 60% acetonitrile in water, TFA condition) to give 33 (43 mg, 4% yield) as a white solid. 1H NMR (400MHz, DMSO-d6 and D2O) δ7.75(dd,J=8.0,3.9Hz,2H),7.28(d,J=8.1Hz,2H),7.13(d,J=6.6 Hz,2H),7.08(d,J=7.6Hz,2H),6.37(s,1H),4.34(s,1H),4.05(q,J=6.7Hz,1H),3.58-3.48(m,4H) ,3.48-3.34(m,4H),2.97(dd,J=9.3,6.2Hz,2H),2.86(dd,J=9.6,5.9Hz,2H),2.38(d,J=6.5Hz,4 H),1.99(dt,J=40.0,6.8Hz,2H),1.84-1.71(m,1H),1.26(d,J=6.6Hz,3H),0.81(t,J=6.1Hz,6H). LCMS:C 37 H 45 Calculated m / z for N7O6: 683.81, Found: 684.65. [M+H] + .

[0333] Example 34. N2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl)benzoyl)-N5-(2-((S)-2-(4-isobutylphenyl)propanamido)ethyl)-L-glutamine (34)

[0334] [ka]

[0335] [ka]

[0336] Step 1: EDCI (3.71 g, 19.42 mmol) was added to a mixture of acid 8-1 (2.00 g, 9.69 mmol), amine 9-1 (2.33 g, 14.54 mmol), and DMAP (238 mg, 1.95 mmol) in DCM (20 mL), and the mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with DCM (50 mL) and washed with 1N HCl (50 mL x 2) and brine (50 mL x 2). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to give the title compound 34-1 (3.2 g, crude) as a white solid, which was used in the next step without further purification. LCMS: C 20 H 32 Calculated m / z for N2O3: 348.49, Found: 397.47. [M+Na] + .

[0337] Step 2: To a solution of compound 34-1 (3.2 g, crude) in EtOAc (30 mL), 4N HCl in EtOAc (30 mL) was added and the mixture was stirred at room temperature for 1 h to allow the complete formation of a white precipitate. The mixture was diluted with EtOAc (30 mL) and filtered. The precipitate was washed with EtOAc and dried to give the title compound 34-2 (1.9 g, 68% yield for two steps) as a white solid. LCMS: C 15 H 24 Calculated m / z for NO: 248.37, Found: 249.90. [M+H] + .

[0338] Step 3: EDCI (1.28 g, 6.70 mmol) was added to a solution of DMAP (82 mg, 0.672 mmol), TEA (680 mg, 6.73 mmol), compound 34-2 (1.0 g, 3.51 mmol), and compound 31-1 (1.01 g, 3.32 mmol) in DCM (20 mL) at 0° C. The reaction was stirred at room temperature for 2 h, and the mixture was diluted with DCM (50 mL) and washed with 1N HCl (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to give the title compound 34-3 (953 mg, 53% yield) as a white solid. LCMS: C 29 H 47 m / z calculated for N3O6: 533.71, found: 556.98. [M+Na] + .

[0339] Step 4: Compound 34-3 (700 mg, 1.31 mmol) was dissolved in DCM (7 mL) and TFA (7 mL) and the reaction was stirred at room temperature for 2 h. The mixture was concentrated to give the title compound 34-4 (700 mg, 90% yield) as a pale yellow oil. LCMS: 20 H 31 Calculated m / z for N3O4: 377.49, Found: 378.88. [M+H] + .

[0340] Step 5: A mixture of compound 31-4 (135 mg, 0.452 mmol), DMAP (5 mg, 0.0409 mmol), NHS (78 mg, 0.678 mmol), and EDCI (172 mg, 0.900 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 2 h. After the complete formation of the NHS-activated ester, compound 34-4 (310 mg, 0.630 mmol) and Et3N (230 mg, 2.27 mmol) were added. The mixture was stirred at 30° C. for 12 h and purified by preparative HPLC (eluted with 60% acetonitrile in water, TFA condition) to give 34 (26 mg, 6% yield) as a white solid. 1H NMR (400MHz, DMSO-d6 and D2O) δ7.81(dd,J=18.1,7.9Hz,2H),7.30(dd,J=11.5,7.9Hz,2H), 7.19-7.15(m,2H),7.06-7.02(m,2H),6.36(s,1H),4.33(dd,J=10.2,4.6Hz,1H),3.14-3.0 0(m,4H),2.99-2.94(m,2H),2.86(dd,J=9.3,5.8Hz,2H),2.37(d,J=7.1Hz,2H),2.18(t,J= 7.5Hz, 2H), 2.14-1.86 (m, 2H), 1.82-1.70 (m, 1H), 1.28 (d, J=6.9Hz, 3H), 0.84-0.79 (m, 6H). LCMS:C 35 H 43 Calculated m / z for N7O6: 657.77, Found: 658.86. [M+H] + .

[0341] Example 35. trans-[Pt(DACH)(ox)(OH)(IPBA)] (35, IPBA = 4-(4-iodophenyl)butyric acid) (35)

[0342] [ka]

[0343] [ka]

[0344] Step 1: Hydrogen peroxide (30%, 15 mL) was added dropwise to a suspension of oxaliplatin 35-1 (200 mg, 0.50 mmol) in water (5 ml). After the addition, the reaction mixture was heated to 75° C. and stirred for 5 h. A clear solution was formed and cooled to room temperature. The resulting solution was concentrated. The residue was washed with EtOH and MTBE to give the title compound 35-2 (170 mg, 78% yield) as a yellow solid.

[0345] Step 2: To a solution of acid 1-2 (114 mg, 0.39 mmol) and TBTU (127 mg, 0.39 mmol) in anhydrous DMSO (5 mL) was added TEA (55 uL, 0.39 mmol). The mixture was stirred vigorously at room temperature for 15 min. Compound 35-2 (170 mg, 0.39 mmol) was added and the reaction mixture was stirred at 60° C. for 16 h. The resulting reaction mixture was filtered to remove unreacted solids. The clear solution was purified by reverse-phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, neutral conditions). The desired component was lyophilized overnight to give 35 (50.8 mg, 18% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ7.59(d,J=7.8Hz,2H),6.97(d,J=7.8Hz,2H),2.44(d,J=7.6Hz,2H ),2.15(d,J=7.5Hz,2H),2.06(t,J=17.4Hz,2H),1.68(t,J=7.5Hz,2H),1.56-0.99(m,8H).

[0346] Example 36. trans-[Pt(DACH)(ox)(IPBA)2] (36, IPBA = 4-(4-iodophenylbutyric acid) (36)

[0347] [ka]

[0348] [ka]

[0349] Step 1: A mixture of compound 1-2 (587 mg, 2.03 mmol), TBTU (650 mg, 2.02 mmol), and TEA (205 mg, 2.02 mmol) in DMF (5 mL) was stirred at room temperature under nitrogen for 15 min. To this mixture, compound 35-2 (218 mg, 0.506 mmol) was added in one portion. The resulting reaction was stirred at 60° C. for 16 h, and the mixture was filtered to remove unreacted solids. The clear solution was directly purified by reverse-phase flash chromatography (C18 column, acetonitrile and water, neutral condition) to give 36 (60 mg, 12% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ7.60(dd,J=8.0,3.8Hz,4H),6.97(dd,J=8.2,3.8Hz,4H),2.47(s, 4H), 2.23(q,J=6.6Hz,4H),2.10(d,J=12.9Hz,2H),1.77-1.62(m,4H),1.53-1.03(m,8H). LCMS:C 28 H 34 Calculated m / z for I2N2O8Pt: 975.48, Found: 976.57 [M+H] + .

[0350] Example 37. N-((S)-1-(((R)-1-(6-(2-(2-(2-((S)-2-(4-isobutylphenyl)propanamido)ethoxy)ethoxy)ethyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (37)

[0351] [ka]

[0352] [ka]

[0353] Step 1: To a solution of compound 37-1 (1.0 g, 4.02 mmol) in DMF (10 mL), NaHCO3 (3.38 g, 40.23 mmol), KI (669 mg, 4.03 mmol), and compound 9-2 (3.36 g, 20.11 mmol) were added. The mixture was stirred at room temperature for 4 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to give the title compound 37-2 (1.14 g, 67% yield). LCMS: C 19 H 36 Calculated m / z for N2O8: 420.50, Found: 443.85 [M+Na] + .

[0354] Step 2: A mixture of compound 37-2 (1.14 g, 2.71 mmol) in 4N HCl (EtOAc solution, 10 mL) was stirred at room temperature for 1.5 h. After completion of the reaction, the mixture was concentrated to give title compound 37-3 (1.36 g, crude), which was used in the next step without further purification.

[0355] Step 3: A solution of compound 37-3 (1.36 g, crude), compound 8-1 (670 mg, 3.24 mmol), DIPEA (525 mg, 4.06 mmol), HOBt (360 mg, 2.66 mmol), and EDCI (520 mg, 2.71 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with DCM (100 mL) and washed with water (100 mL x 2) and brine (100 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by reverse-phase flash chromatography (C18 column, eluted with 65% acetonitrile in water, HCl condition). The desired components were lyophilized to give the title compound 37-4 (948 mg, TFA salt, 64% yield for two steps) as a white solid. LCMS: C 27 H 44 Calculated m / z for N2O7: 508.66, Found: 531.73 [M+Na] + .

[0356] Step 4: To a solution of compound 37-4 (945 mg, 1.73 mmol) in EtOH (9.5 mL) and H2O (3 mL) was added NaOH (297 mg, 7.45 mmol) at 0 °C. The reaction was stirred at room temperature for 2 h. The mixture was concentrated and adjusted to pH = 2 with 1N HCl at 0 °C. The aqueous mixture was purified by reverse-phase flash chromatography (C18 column, eluted with 45% acetonitrile in water, HCl condition). The desired component was lyophilized to give the title compound 37-5 (694 mg, 82% yield) as a white solid. LCMS: C 23 H 36 Calculated m / z for N2O7: 452.55, Found: 453.52 [M+H] + .

[0357] Step 5: A mixture of compound 37-5 (100 mg, 0.204 mmol) and compound 9-7 (85 mg, 0.22 mmol) in toluene (3 mL) was stirred at 120° C. for 4 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to give 37 (97 mg, 59% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.92(d,J=7.8Hz,1H),8.84(s,1H),8.71(s,1H),7.95(s,1H),7.37(d,J=9.3H) z,1H),7.29-7.12(m,7H),7.03(d,J=6.9Hz,2H),4.66(d,J=6.2Hz,1H),4.28(d,J=17.6Hz,1H),4.02(dd,J=32.9,1 7.2Hz,2H),3.68(t,J=18.8Hz,3H),3.46(s,3H),3.39-3.25(m,4H),3.16(s,2H),3.09-3.00(m,2H),2.37(s,2H),1 .76(s,1H),1.55(s,1H),1.41(t,J=12.4Hz,1H),1.27(d,J=6.3Hz,3H),1.23-1.16(m,1H),0.82(d,J=6.5Hz,12H). LCMS:C 42 H57 Calculated m / z for BN6O9: 800.76, Found: 801.76 [M+H] + .

[0358] Example 38. (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl (S)-2-(4-(2-(methyl-d3)propyl-3,3,3-d3)phenyl)propanoate (38)

[0359] [ka]

[0360] [ka]

[0361] Step 1: A mixture of compound 22-11 (30.0 mg, 0.141 mmol), EDCI (80.0 mg, 0.419 mmol), and HOBt (20.0 mg, 0.148 mmol) in anhydrous DMF (2 mL) was stirred at room temperature for 5 min. Compound 1-1 (85 mg, 0.216 mmol) was added, and the mixture was stirred at room temperature for 16 h. The reaction mixture was partitioned between EtOAc (30 mL) and water (20 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by pre-TLC (5% MeOH in DCM) to give the title compound 38 (50 mg, 60% yield) as an off-white solid. 1H NMR(500MHz,DMSO-d6)δ8.18(d,J=9.1Hz,1H),7.90(d,J=2.5Hz,1H),7.51(dd,J=9.1,2. 5Hz,1H),7.37(d,J=7.9Hz,2H),7.32(s,1H),7.20(d,J=7.8Hz,2H),5.43(s,2H),5.32(s, 2H),4.15(q,J=7.1Hz,1H),3.16(q,J=7.6Hz,2H),2.45(d,J=7.2Hz,2H),1.85(ddt,J=28 .9,14.6,7.2Hz,3H),1.57(d,J=7.1Hz,3H),1.27(t,J=7.6Hz,3H),0.88(t,J=7.3Hz,3H). LCMS:C 35 H 30 Calculated m / z for D6N2O6: 586.72, Found: 588.13 [M+H] + .

[0362] Example 39. Cellular IC50 in three tumor cell lines Cell inhibition was determined in three assays: U87MG, A549, and MC38. Cells were recovered in appropriate medium supplemented with 10% fetal bovine serum and 100 U / mL sodium penicillin G and maintained in a cell incubator (37°C, 5% CO2). Before testing, cells in culture dishes were rinsed with phosphate buffer and detached with trypsin. The cell number was diluted and adjusted with culture medium, and the cell suspension was added to a 96-well cell plate. The cells were maintained overnight in the incubator. In the T0 control plate, 100 μL of CellTiter-Glo reagent was added to the cells, balanced at room temperature for 10 minutes, and the chemiluminescence value was read as expected.

[0363] In the test plate, the compounds were dissolved in the corresponding solvent and diluted gradiently. The diluted compound solutions were added to the 96-well plate with cells. After 72 h of incubation, the cells were added with 100 μL of CellTiter-Glo reagent, balanced at room temperature for 10 min, and read for chemiluminescence values. IC50 was calculated using the GraphPad Prism software package (Prism6 for Windows, version 6.0, GraphPad Software Inc., San Diego, CA). IC50 for the three cell assays 50 is shown in Table 1.

[0364] [Table 4]

[0365] Example 40. Chemical stability in PBS (pH 7.4) The chemical stability assay was carried out according to the following procedure. 1. Test compound spiking solutions: 1 mM test compound spiking solution A: To 10 μL of 10 mM test compound stock solution, add 90 μL of DMSO. 2. Add 396 μL of buffer into the tubes designated for different time points. Pre-warm the samples at 37° C. for 10 minutes. 3. To wells containing 396 μL of buffer and designated for 0 minutes (or 15, 45, 90, 120 minutes), add 4 μL of Spike Solution A and then begin the timing countdown. 4. At each time point, add 1200 μL of ACN containing IS to the tubes. 5. The samples are centrifuged at 10,000 rpm for 5 minutes and then 100 μL of the supernatant is prepared for LC-MS / MS analysis.

[0366] [Table 5]

[0367] Example 41. Pharmacokinetics Pharmacokinetic parameters were determined in male CD-1 mice (Shanghai Jihui Laboratory Animal Care Co., Ltd., 6-8 weeks old). Animals were maintained under a 12-h light / dark cycle. Animals had free access to food and water during the study.

[0368] Intravenously, compounds were formulated as solutions (5% DMSO + 10% Solutol HS15 + 85% (20% HP-β-CD in water)) at a dose volume of 5 mL / kg and administered via the tail vein. Subcutaneously, compounds were formulated as solutions (5% DMSO + 10% Solutol HS15 + 85% (20% HP-β-CD in water)) at a dose volume of 10 mL / kg or as solutions (0.5% MC + 1% Pluronic F68 in water) at a dose volume of 50 μL / mouse and administered via subcutaneous puncture.

[0369] Semi-timed blood samples (approximately 110 μL) were collected from animals at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours for the IV group, and 0.5, 2, 4, 8, 24, 48, and 96 hours for the SC group. Within 15 minutes after sample collection, samples were kept on ice for 15 minutes or less before centrifugation (2000 g, 5 minutes, 4° C.). Plasma was flash frozen in dry ice and then transferred to a −70° C. freezer for long-term storage until LC-MS / MS analysis.

[0370] Tissues from the SC group were collected at 0.5, 2, 4, 8, 24, 48, and 96 hours. After the animals were anesthetized and exsanguinated, tissue samples (including muscle and skin) were collected and weighed, then flash frozen in liquid nitrogen and further stored at -70°C for long-term storage until LC-MS / MS analysis. Tissue samples were homogenized under freezing conditions.

[0371] PK parameters were generated from the LC-MS / MS data using Phoenix WinNonlin 8.2 software.

[0372] [Table 6]

[0373] Applicant's disclosure is described herein in a preferred embodiment with reference to the drawings, in which like numbers represent the same or similar elements. Throughout this specification, reference to "one embodiment," "an embodiment," or similar language means that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment of the invention. Thus, appearances of the phrases "in one embodiment," "in an embodiment," and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

[0374] The described features, structures, or characteristics of the applicant's disclosure may be combined in any suitable manner in one or more embodiments. In the description herein, numerous specific details are recited to provide a thorough understanding of the embodiments of the present invention. However, one skilled in the art will recognize that the applicant's compositions and / or methods may be practiced without one or more specific details, or with other methods, components, materials, etc. In other instances, well-known structures, materials, or operations have not been shown or described in detail to avoid obscuring aspects of the present disclosure.

[0375] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the content clearly dictates otherwise.

[0376] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this disclosure, the preferred methods and materials are described herein. The methods described herein can be carried out in any order that is logically possible, in addition to the specific order disclosed.

[0377] Incorporation by Reference In this disclosure, references and citations are made to other documents, such as patents, patent applications, patent publications, journals, books, papers, web content, etc. All such documents are incorporated herein by reference in their entirety for all purposes. Any material or portion thereof that is incorporated herein by reference but that conflicts with existing definitions, descriptions, or other disclosure materials explicitly set forth herein is incorporated only to the extent that no conflict occurs between the incorporated material and this disclosure material. In the event of a conflict, the conflict shall be resolved in favor of this disclosure as the preferred disclosure.

[0378] Equivalent The representative examples are intended to help explain the invention, and are not intended to limit the scope of the invention, and should not be construed as such. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the entire contents of this document, including the examples and references to the scientific and patent literature contained herein. The examples contain important additional information, exemplification and guidance that can be adapted to the practice of the invention in its various embodiments and equivalents thereof.

Claims

1. Structural formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable form or isotopic derivative thereof, During the ceremony, X is CHR or NR; R is H or alkyl; A compound, or a pharma- ceutically acceptable form or isotopic derivative thereof. 【Request 2】 【Chemistry 2】 2. The compound of claim 1, or a pharma- ceutically acceptable form or isotopic derivative thereof, selected from:

3. Structural formula (II): 【Chemistry 3】 or a pharma- ceutically acceptable form or isotopic derivative thereof. 【Request 4】 【Chemical 4】 4. The compound of claim 3, or a pharma- ceutically acceptable form or isotopic derivative thereof, selected from:

5. Structural formula: 【Chemistry 5】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

6. Structural formula (III): 【Chemistry 6】 or a pharma- ceutically acceptable form or isotopic derivative thereof, wherein n is an integer selected from 2 to 8; A compound, or a pharma- ceutically acceptable form or isotopic derivative thereof. 【Request 7】 【Chemical 7】 7. The compound of claim 6, selected from:

8. Structural formula (IV): 【Chemistry 8】 or a pharma- ceutically acceptable form or isotopic derivative thereof, wherein n is an integer selected from 2 to 10; A compound, or a pharma- ceutically acceptable form or isotopic derivative thereof. 【Request 9】 【Chemistry 9-1】 【Chemistry 9-2】 【Chemistry 9-3】 9. The compound of claim 8, or a pharma- ceutically acceptable form or isotopic derivative thereof, selected from:

10. Structural formula (V): 【Chemistry 10】 or a pharma- ceutically acceptable form or isotopic derivative thereof, During the ceremony, R ２ is H or P(=O)R ２ and R is alkyl; L is a single bond, or 【Chemistry 11】 is a group selected from k is an integer selected from 0 to 4; A compound, or a pharma- ceutically acceptable form or isotopic derivative thereof.

11.

12. 11. The compound of claim 10, or a pharma- ceutically acceptable form or isotopic derivative thereof, selected from:

12. Structural formula (VI): 【Chemistry 13】 or a pharma- ceutically acceptable form or isotopic derivative thereof, During the ceremony, R ２ is H or P(=O)R ２ and R is alkyl; L is a single bond, or 【Chemistry 14】 is a group selected from k is an integer selected from 0 to 4; A compound, or a pharma- ceutically acceptable form or isotopic derivative thereof.

13. The structure: 【Chemistry 15】 13. The compound of claim 12, or a pharma- ceutically acceptable form or isotopic derivative thereof, having the formula:

14. Structural formula: 【Chemistry 16】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

15. Structural formula: 【Chemistry 17】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

16. Structural formula: 【Chemistry 18】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

17. Structural formula: 【Chemistry 19】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

18. Structural formula: 【Chemistry 20】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

19. Structural formula: 【Chemistry 21】 or a pharma- ceutically acceptable form or isotopic derivative thereof.

20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19 and a pharma- ceutically acceptable excipient, carrier, or diluent.

21. 21. The pharmaceutical composition of claim 20, which is effective to treat or reduce cancer or a related disease or disorder.

22. 22. A unit dosage form comprising the pharmaceutical composition of claim 20 or 21.

23. 20. A method of treating or ameliorating a disease or condition comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound according to any one of claims 1 to 19 and a pharma- ceutically acceptable excipient, carrier, or diluent.

24. 24. The method of claim 23, wherein the disease or condition is cancer or a related disease or condition.

25. 20. Use of a compound according to any one of claims 1 to 19 for treating or ameliorating a disease or condition.

26. 20. Use of a compound according to any one of claims 1 to 19 together with a pharma- ceutically acceptable excipient, carrier or diluent in the preparation of a medicament for treating or ameliorating a disease or disorder.

27. 27. The use according to claim 25 or 26, wherein the disease or condition is cancer or a disease or condition related thereto.

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