Stable heavy isotopes in amide functional groups and uses thereof

Abstract

Isotopically enriched compounds comprising a stable, heavy isotope-enriched amide functional group for modulating the pharmacokinetic profile, metabolic profile, and / or delivery efficiency of a drug or prodrug, and their therapeutic or prophylactic efficacy and / or adverse effects are provided. Also provided is the use of drugs and prodrugs comprising isotopically enriched amides for the treatment or prevention of disease states and conditions.(I)

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Application No. 62 / 994,378, filed March 25, 2020, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This disclosure relates to isotopically enriched compounds containing stable heavy isotope-enriched amide functional groups for modulating the pharmacokinetic profile, metabolic profile, and / or delivery efficiency of compounds (e.g., drugs or prodrugs), and to their therapeutic and prophylactic uses. Background Technology

[0004] Amides, also known as acid amides, are compounds containing the functional group R. m E(O) n NR 1 R 2 Compounds in which R, R 1 and R 2 It is either hydrogen (H) or an organic group. The most common amide is formamide (organic amide), where m is 1, E is carbon (C), and n is 1. Many other important types of amides are known, including phosphoramides (e.g., where m is 1, E is phosphorus (P), and n is 2, and related compounds) and sulfonamides (e.g., where m is 1, E is sulfur (S), and n is 2, and related compounds) (see IUPAC, Compendium of Chemical Terminology, 2nd edition (“Gold Book”), 1997). Another class of amides is phosphonamide.

[0005] Structurally, the amide bonds in the compound can be represented as follows:

[0006]

[0007] These are nailamide, phosphoramide, phosphonamide, and sulfonamide; among which R and R 1 R 2 and R 3 It is hydrogen or an organic group such as alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl or a combination thereof, without or with other substituents.

[0008] In conventional nomenclature, the term "amide" is added to the stem of the acid name. For example, an amide derived from acetic acid is named acetamide (CH3CONH2). IUPAC recommends naming such a compound ethanamide, but this and related formal names are rarely encountered. When an amide is derived from a primary or secondary amine, the substituent on the nitrogen atom is indicated first in the name. Thus, the amide formed from dimethylamine and acetic acid is N,N-dimethylacetamide (CH3CONMe2). Cyclic amides are called lactams; they must be secondary or tertiary amides. (The last sentence appears to be incomplete and possibly refers to a different topic.) 1 R 2 and –SO2NR 1 R 2 The functional groups that make up the product are phosphonamide and sulfonamide (see IUPAC, Nomenclature of Organic Chemistry, Sections A, B, C, D, E, F and H, Pergamon, Oxford, 1979. Rules C-821: Amides and Imides).

[0009] Amides are important functional groups found in many types of drugs, such as local anesthetics, antiarrhythmics, and others. Amides are also key linking motifs in protein and peptide drug products (DeRuiter, J., Principles of Drug Action 1, Spring 2005, Amides). Furthermore, many drugs developed in the last century were amine prodrugs, highlighting their importance in the medical field. It is now widely accepted that amine prodrugs play a crucial role in drug targeting, and they are often the initial compounds that deliver drugs to their target sites in a stable form. Amine prodrugs are typically classified by the molecular bonding of the nitrogen atom to nearby atoms. Amine-containing prodrugs exhibit a variety of basic functional groups and linkages, such as amide prodrugs, azo-linked prodrugs, and lipopeptide prodrugs (Chandy, A., et al., Med. Chem. Drug Discov., 2013, 4(2), 108-126; Simplício, AL, et al., Molecules, 2008, 13, 519-547).

[0010] For example, many anticancer agents have amide bonds (see, for example, Mohammed, YHE and Khanum, SA, Int. J. of Pharma and Bio Sci. (2018), 9(2), 94-124; Wang, B, et al., Drug Delivery, 2nd ed., 2016, 475-502). As a specific example, Alpelisib (formerly known as BYL719, or N-(4-methyl-5-(2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridyl)-2-thiazolyl)aminocarbonyl-L-prolylamide) is an α-specific PI3K (phosphoinositol 3-kinase) inhibitor investigated clinically for the treatment of some cancers (also described in U.S. Patent Nos. 8,476,268 and 8,227,462). Recent studies in the SOLAR-1 trial targeting PIK3CA alterations in metastatic breast cancer have demonstrated the effectiveness of apegliflozin (PIQRAY). TM The efficacy of this compound in treating breast cancer has been investigated. Clinical studies have also been conducted in patients with advanced solid tumors (Ando, ​​Y., et al., Cancer Science, 2019, 110:1021-1031).

[0011] Another type of drug containing amide functional groups is RNA interference (RNAi) therapeutics. More than 20 RNAi-based therapies are currently in clinical trials, and positive results from these trials are encouraging further efforts to develop clinically relevant RNAi treatments (Bobbin & Rossi, Annu. Rev. Pharmacol. Toxicol., 2016, 56:103–22). Such novel therapeutic agents under development include siRNA conjugates for treating diseases (Weingaertner & Bethge, (2019), WO2019193144; Bethge, et al., (2019), WO2019193189; Zhang, et al., (2019), WO2019105437; Zhang, et al., (2019), WO2019105414; Nair, et al., (2019), WO2019217459; Nair, et al., (2015) US2015 / 0196655; Muthiah, et al., (2015) WO2015006740). Synthetic small interfering RNA (siRNA) can suppress the expression of pathogenic genes through post-transcriptional gene silencing mediated by the endogenous RNA interference (RNAi) pathway. siRNAs possess enormous therapeutic potential, but efficient delivery to target cells or organs remains a challenge. Covalent conjugation of small molecule compounds to siRNAs has been used to avoid the side effects associated with delivery systems based on non-viral vectors, particles, or excipients. For example, conjugation of cholesterol and other lipophilic moieties to siRNAs can lead to widespread biodistribution and gene silencing in a variety of tissues, including the liver (Nair, JK; et al., J. Am. Chem. Soc. 2014, 136, 16958-16961; Wolfrum, C.; et al., Nat. Biotechnol. 2007, 25, 1149). In addition, well-characterized biantennary and triantennary GalNAc ligands can be reprogrammed to promote covalent conjugation with siRNA (Khorev, O.; et al., Bioorg. Med. Chem. 2008, 16, 5216; Rensen, PCN; et al., J. Med. Chem. 2004, 47, 5798; Valentijn, ARPM; et al., Tetrahedron, 1997, 53, 759; Manoharan, M.; et al., WO 2009073809, 2009).

[0012] One of the key reactions involving carboxylic amides (also known as carboxylic amides and formamides) and peptides is hydrolysis. Hydrolysis can be acid-catalyzed, base-catalyzed, or enzyme-catalyzed, and typically yields the corresponding carboxylic acid and amine. After the C=O double bond is converted to a CO single bond, hydrolysis involves a tetrahedral intermediate at the carbon center, as shown in acid-catalyzed hydrolysis:

[0013]

[0014] The rate determination step is step 2, in which the C=O double bond is cleaved into a CO single bond by the π bond, which has a direct impact on the rate of amide bond cleavage.

[0015] Foreign substances, including compounds and other therapeutic agents, are frequently metabolized to facilitate their excretion from the body. For example, various enzymes (such as cytochrome P450 enzymes, esterases, proteases, reductases, dehydrogenases, transaminases, and monoamine oxidases) can react with compounds and therapeutic agents, catalyzing their conversion into more polar metabolites for renal excretion. The resulting metabolites can have significantly different pharmacokinetic, pharmacodynamic, and acute and long-term toxicity profiles relative to the parent compound.

[0016] In some cases, such metabolic reactions can involve oxidizing carbon-hydrogen bonds to carbon-oxygen or carbon-carbon π bonds. The strength of a carbon-hydrogen bond is proportional to the absolute value of its ground-state vibrational energy. This vibrational energy depends on the mass of the atoms forming the bond and increases with the mass of one or both atoms forming the bond. Deuterium is a stable and non-radioactive isotope of hydrogen, with a mass approximately twice that of protium, the most common isotope of hydrogen. Since the mass of deuterium (D) is approximately twice that of protium (… 1 The carbon-deuterium (C-H) bond is twice the mass of the carbon-deuterium (C-H) bond, therefore the carbon-deuterium (CD) bond is stronger than the corresponding carbon-deuterium (C-H) bond. 1 H) bond. If during the rate-determining step of a metabolic reaction, C- 1 If the H bond is cleaved, then replacing the protium with deuterium can lead to a decrease in the reaction rate. In this way, D can have a significant isotopic effect.

[0017] Deuteration of drugs has been shown to improve pharmacokinetics and pharmacodynamics. For example, the deuterated drug SD-809 (bubenazine-d6 or deuterated bubenazine) has been used to treat Huntington's disease and tardive dyskinesia. The deuterated form of sorafenib, donafenib, has been clinically tested for the treatment of colorectal and thyroid cancer. Isotope enrichment can potentially affect the metabolism of therapeutic agents, the release, absorption, and / or clearance of prodrugs and derivatives, significantly altering the pharmacokinetic profile of the drug.

[0018] An isotope is an atom that has the same number of atoms but different mass numbers. Many elements have more than one stable isotope. Common elements with more than one stable isotope include hydrogen, boron, carbon, nitrogen, oxygen, chlorine, magnesium, silicon, sulfur, potassium, calcium, titanium, chromium, iron, nickel, zinc, selenium, and bromine. Some elements have two or more isotopes with similar natural abundances. For example, bromine has two stable isotopes: 79 Br and 81 Br, where the natural abundances of these two isotopes are 50.69% and 49.31%, respectively. Similarly, the natural abundances of stable chlorine isotopes are: 35 Cl is 76% and 37 Cl is 24%.

[0019] Other elements have more than one stable isotope, possessing a major isotopic form and one or more minor isotopic forms. For such elements, isotopes are distinguished by two factors: (1) their atomic weight, which is attributed to the number of neutrons in the nucleus, and (2) their abundance in nature. For hydrogen, a common stable isotope is the proton (H or H). 1 H) and deuterium (D or 2 Hydrogen (H) is the dominant isotope in nature, with only 0.0115% being D (H). 2 For oxygen, there are three stable isotopes: 16 O、 17 O and 18 O. For 16 O、 17 O and 18 O, the natural occurrence rates of these three stable oxygen isotopes are generally 99.756%, 0.039%, and 0.205%, respectively. From 16 O to 18 O, the number of neutrons increased by 2 (from 8 to 10), resulting in a 12.5% ​​increase in atomic weight. Because 16 O and 18 The difference in atomic weight between O atoms is smaller than the difference in atomic weight between D and H atoms, therefore in some cases, using 18 O substitution 16 The isotopic effect of O on, for example, the metabolism of compounds can be smaller than the isotopic effect of replacing H with D.

[0020] As one of the stable isotopes 18 O has been widely used in many fields of medical science and general health, including as a component of human physiology. 18 O tracer. It has been used in drug metabolism and pharmacokinetics (DMPK) studies. 18 O substitution 16O is used to trace drug compounds and their metabolites after administration, in order to achieve sensitive, accurate and rapid results. 18 O is also used in research on energy metabolism and consumption. For example, it can be used to study energy metabolism and consumption. 18 O water ("H2") 18 (O" or "oxygen-18 water"). If H2 18 The replacement of hydrogen atoms in O with deuterium isotopes provides D2. 18 O (often referred to as "doubly-labeled water" or "DLW"). DLW provides a rapid and accurate method for determining energy expenditure in animals and humans; this method is based on the premise that after a certain dose of DLW, the two isotopes reach equilibrium with total body water (TBW) and are subsequently eliminated from the body in different forms. Deuterium leaves the body as water, while... 18 O2 leaves the body in the form of water and carbon dioxide, thus allowing energy metabolism to be determined by measuring isotopic excretion rates. [Further details about studies are also needed.] 18 The effects of O on the growth and reproduction of C-57 mice (Wolf, D.; Cohen, H.; Meshorer, A.; Wasserman, I.; Samuel D., Stable Isot., Proc. Int. Conf., 3rd (1979), 353-60). In these studies, O was highly enriched in the respiration of paired C-57 mice for three consecutive generations. 18 Mice were kept in an O2 atmosphere for 112 days. No significant changes were detected in organs, and mice survived and reproduced normally, even when 60% of their oxygen was depleted. 18 The same applies when O is substituted. Studies have shown... 18 The conclusion that O is harmless to mice. In fact, 18 O is safe and well-tolerated in the human body; it can be estimated that for an average person weighing 60 kg, the body contains approximately 40 kg of O composed of all three stable isotopes. 16 O、 17 O and 18 O) is composed of oxygen, of which about 80g is 18 O. Similarly, for other elements (e.g., C and N) that have more than one stable isotope, stable isotopes with one major isotopic form and one or more minor isotopic forms can be found to exist naturally at their native abundance levels and are generally safe and well tolerated. Summary of the Invention

[0021] One object of the present invention is to provide an amide moiety having an improved amide bond cleavage rate to improve the pharmacokinetic profile of drugs and prodrugs containing amide bonds, thereby modulating their therapeutic and preventative effects and / or adverse effects.

[0022] This invention is based, at least in part, on the inventors' understanding of the following: stable heavy isotopes ( 17 O and / or 18 O; 13 C; and / or 15 N) and naturally abundant isotopes ( 16 O、 12 C and 14 Compared to N), it can alter the amide bond cleavage rate, thereby improving the pharmacokinetic profile of drugs and prodrugs containing amide bonds, in order to modulate the therapeutic, preventative, and / or adverse effects of drugs and prodrugs containing amide bonds.

[0023] Therefore, stable heavy isotopes ( 17 O and / or 18 O; 13 C; and / or 15 N) replaces naturally abundant oxygen, carbon, or nitrogen isotopes ( 16 O、 12 C or 14 N) can produce an isotopic effect on the cleavage rate of amide bonds. To avoid being limited by theory, it is considered that stable heavy isotopes ("N") can be used. * O" or "O * ",represent 17 O and / or 18 O; "*C" or "C*" represents 13 C; and / or "*N" or "N*", representing 15 N) Substitution 16 O、 12 C and / or 14 Nitrogen (N) can alter the amide bond cleavage rate. This change can improve the pharmacokinetic profiles of drugs and prodrugs containing amide bonds, and thus modulate their therapeutic, prophylactic, and / or adverse effects. Notably, the carboxylamide center directly involves three atoms: oxygen, carbon, and nitrogen, and therefore, isotopes of oxygen and / or carbon and / or nitrogen can directly affect the amide bond cleavage rate, and thus directly influence the therapeutic, prophylactic, and / or adverse effects of drugs and prodrugs containing amide bonds.

[0024] Therefore, pharmaceutical and / or prodrug compounds containing amide bonds are provided, having one or more carboxylamide functional groups (C(=O)-N or ...) in the pharmaceutical and / or prodrug compound. ( ) to replace one or more restable isotopes that are naturally occurring isotopes.

[0025] In the first broad aspect, compounds of formula I or their pharmaceutically acceptable salts, esters, hydrates, chelates, or solvates are provided:

[0026]

[0027] Where n is an integer selected from 1 to 5; R refers to the isotope-enriched amide functional group (also referred to as "isotope-enriched amide" and "isotope-enriched amide group" in this paper); R is the organic part; R 1 and R 2 Independently, it is a hydrogen or organic moiety; and the structure of Formula I comprises, or is contained in, a drug or prodrug compound containing at least one formamide bond; provided that -NR 1 R 2 The part is not the 3-sulfo-1-propylamino moiety.

[0028] The terms “isotope enrichment” and “heavy isotope enrichment” used in this article are used interchangeably to refer to the presence of one or more stable heavy isotopes ( 18 O、 17 O、 13 C and / or 15 Enrichment of N). It should be understood that when two or more atoms are enriched in a compound, these atoms may be enriched with the same or different isotopes; and in Formula I, when n>1, multiple amide groups may be located at different positions of the R group. For example, an amide group may be enriched with two different isotopes of the same element, or alternatively enriched with isotopes of two different elements. Many such combinations and arrangements are possible. When n is greater than 1, each amide group may be linked to the R group at the same or different positions of R; or either of the two amide groups may be linked by a peptide bond (i.e., in the manner in which amino acids are linked in a peptide).

[0029] In one embodiment, a compound of formula I or a pharmaceutically acceptable salt, ester, hydrate, chelate, and / or solvate thereof is provided, wherein the isotope-enriched amide functional group is enriched with one or more stable heavy oxygen isotopes (…). 18 O、 17 O, or 18 O and 17 (a mixture of O); and R, R 1 and R 2 The limitations are the same as those mentioned above.

[0030] In another embodiment, a compound of formula I or its pharmaceutically acceptable salt, ester, hydrate, chelate, and / or solvate is provided, wherein the isotope-enriched amide functional group is enriched with one or more stable heavy carbon isotopes (13 C); and R, R 1 and R 2 The limitations are the same as those mentioned above.

[0031] In another embodiment, a compound of formula I or its pharmaceutically acceptable salt, ester, hydrate, chelate, and / or solvate is provided, wherein the isotope-enriched amide functional group is enriched with one or more stable diazonium isotopes ( 15 N); and R, R 1 and R 2 The limitations are the same as those mentioned above.

[0032] In one embodiment, the isotope-enriched amide is enriched with a single isotope of an element, such as... 18 O enrichment, or 17 O enrichment, or 13 C enrichment, or 15 N-enriched.

[0033] In another embodiment, the isotope enrichment amide enriches two or more isotopes of one or more elements, for example, is 17 O and 18 O enrichment, or 18 O and 13 C enrichment, or 18 O and 15 N enrichment, or 13 C and 15 N-enriched.

[0034] In some implementations of Equation I, n is an integer selected from 1 to 3.

[0035] In one implementation of Equation I, n is 1.

[0036] In some embodiments of Formula I, the R group is an alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or heterocyclic group.

[0037] In one embodiment of Formula I, the R group is an organic moiety selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and heterocyclic, wherein R is substituted or unsubstituted. In one embodiment of Formula I, the R group is alkyl, cycloalkyl, heterocyclic, or heterocyclic alkyl. In another embodiment of Formula I, the R group is alkyl, cycloalkyl, or heterocyclic alkyl. In another embodiment of Formula I, the R group is cycloalkyl or heterocyclic alkyl. In another embodiment of Formula I, the R group is heterocyclic alkyl. It should be understood that in all such embodiments of Formula I, the R group can be substituted or unsubstituted without limitation.

[0038] In one embodiment of Formula I, the R group is not a fused aromatic group, such as not a fused aryl or fused heteroaryl group. In another embodiment of Formula I, the R group is not a 2-substituted 1H-indazole-7-yl group.

[0039] In some implementations of Formula I, R 1 and R 2 Independently, it is hydrogen and a moiety selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heterocyclic, alkoxy, acyl, and alkylthio. In one embodiment of Formula I, R 1 It is hydrogen and R 2 It is alkyl, alkylthio, or aryl. In one embodiment of Formula I, R 1 It is hydrogen and R 2 It is an alkyl group. In one embodiment of formula I, R 1 It is hydrogen and R 2 It is C 1-6 Alkyl group. In another embodiment, R 1 and / or R 2 It is a protecting group selected from acyl, carbonyl, thiocarbonyl, and carbamoyl groups. In another embodiment, R 1 It is hydrogen and R 2 It is a protecting group selected from acyl, carbonyl, thiocarbonyl, and carbamoyl groups. It should be understood that in all such embodiments of Formula I, R... 1 and R 2 It can be substituted or unsubstituted, without restriction.

[0040] In one embodiment of Formula I, R 1 and R 2 Both are hydrogen.

[0041] In one specific embodiment of Formula I, R is a substituted or unsubstituted alkyl, cycloalkyl, or heterocycloalkyl group; R 1 and R 2 Both are hydrogen; and n is 1.

[0042] In some embodiments, isotope-enriched amides, as described herein, are provided, wherein the amide is part of a drug or prodrug. Amides are important functional groups present in many types of pharmaceutical compounds (e.g., local anesthetics, antiarrhythmics, etc.) and can also be key linking moieties in protein and peptide drug products (DeRuiter, J., Principles of Drug Action 1, Spring 2005, Amides). A large number of pharmaceutical compounds contain at least one amide bond or amine moiety, and many drugs developed in the last century are amine prodrugs.

[0043] In some embodiments, the compound of the present invention is an isotopically enriched apelelis, for example... 18 O-Apeliximab. Apeliximab is an amide-containing drug with the following structure:

[0044]

[0045] It has an amide functional group and a proline amide group (derived from the proline moiety, as shown on the right side of the compound). The compound has two oxygen atoms, one (O1) in the simple amide group and the other (O2) in the urea fragment.

[0046] It has been shown that apelis is extensively metabolized at the proline amide group to form the corresponding proline derivative, namely a carboxylic acid (hereinafter referred to as James, A. et al., Cancer Chemother. Pharmacol., (2015), 76(4), 751-760). Unfortunately, this acidic metabolite (“M4”) is biologically inactive.

[0047]

[0048] In one embodiment, an isotope-enriching amide as described herein is provided, wherein the isotope-enriching amide is contained in apeliximab. In one embodiment, a compound of formula I or its pharmaceutically acceptable salt, ester, hydrate, chelate, and / or solvate is provided, wherein R is N-((4-methyl-5-(2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridinyl)-2-thiazolyl)aminocarbonyl)-pyrrolidine-2-yl, or the following portion:

[0049]

[0050] And R 1 and R 2 It is hydrogen or a protecting group, thus providing apelix derivatives with isotopically enriched amide functional groups. Since the amide carbonyl group in apelix is ​​a C=O bond, isotopically enriched apelix can be achieved at the C1 carbon (e.g., by using...). 13 C replaces C1- carbon, that is 13 C1-apelis), at O1 oxygen (e.g., by using... 18 O or 17 O replaces O1 oxygen, that is 18 O1-Apeliximab 17 O1-apelis) or at both C1 and O1 (e.g., O1-apelis) or at both C1 and O1 (e.g., 13 C1 18 O1-Apeliximab 13 C1 17O1-apelis is an isotopically enriched compound. Therefore, in one embodiment, the compound of the present invention is selected from... 13 C1-Apeliximab 18 O1-Apeliximab 17 O1-Apeliximab 13 C1 18 O1-Apelixi and 13 C1 17 O1-Apeliximab

[0051] In one implementation, the compound of formula I is 18 O1-Apeliximab (also referred to as Apeliximab in this article) 18 O1).

[0052] In another embodiment, a compound of formula I is provided as a compound of formula II, or a pharmaceutically acceptable salt, ester, hydrate, chelate, or solvate thereof:

[0053]

[0054] in R 1 and R 2 As specified above.

[0055] In some embodiments, the isotopes in the compound of formula II are enriched with amide functional groups or It is enriched by a single isotope, for example, 18 O enrichment, or 17 O enrichment, or 13 C enrichment, or 15 N is enriched; and R is enriched. 1 and R 2 The limitations are the same as those mentioned above.

[0056] In another embodiment, the isotope enrichment of the amide functional group in the compound of formula II or It is an amide functional group enriched with two or more heavy isotopes of one or more elements, for example... 17 O and 18 O enrichment, or 18 O and 13 C enrichment, or 18 O and 15 N enrichment, or 13 C and 15 N enrichment, or 17 O and 13 C enrichment, or 17 O and 15 N enrichment.

[0057] In one embodiment, compounds having isotopically enriched amide functional groups include niraparib and mefuparib, both of which are PARP inhibitors used to treat cancer, and the organic parts R in Formula I are respectively: [details about niraparib and mefuparib]. And for Meifupire is Therefore, the corresponding isotope-enriched drugs have the following structures: (Nirapani) (Mefuperil), in which the compound contains It is an amide functional group enriched with two or more heavy isotopes of one or more elements, for example... 17 O and 18 O enrichment, or 18 O and 13 C enrichment, or 18 O and 15 N enrichment, or 13 C and 15 N enrichment, or 17 O and 13 C enrichment, or 17 O and 15 N enrichment.

[0058] In an alternative embodiment, the compound having an isotopically enriched amide functional group is not nirapanib, i.e., R in formula I is not... In one specific implementation, the compound having an isotope-enriched amide functional group is not 14 C-nirapanib. In one specific embodiment, the compound having an isotopically enriched amide functional group is not... 18 O-niraparib. In one specific embodiment, the compound having an isotopically enriched amide functional group is not... 13 C-nirapanib. In one specific embodiment, the compound having an isotopically enriched amide functional group is not... 17 O-Nirapani.

[0059] Another example of an amide-containing drug that may contain isotope-enriched amide functional groups as described herein is Selexipag, a drug developed by Actelion for the treatment of pulmonary arterial hypertension (PAH). Celecoxib and its active metabolite ACT-333679 (or MRE-269, free carboxylic acid) are agonists of the prostacyclin receptor, leading to vasodilation in the pulmonary circulation (Sitbon, O.; Morrell, N., Eur. Respir. Rev., 2012, 21(126):321–327). Celecoxib is a prodrug that is hydrolyzed to release its active substance, as follows:

[0060]

[0061] Another example of an amide-containing drug that may contain isotopically enriched amide functional groups as described herein is midodrine, an N-glycyl derivative of Desglymidodrine. The former is deprotected by a peptidase to obtain the latter, as follows:

[0062]

[0063] Another example of an amide-containing drug or prodrug is an NSAID prodrug (e.g., see Husain A., et al., Sch. Acad. J. Pharm., 2015; 4(3):145-152), which is represented by the following general structure:

[0064]

[0065] NSAIDs include, but are not limited to, acetylclofenac, diclofenac, fenbufen, indomethacin, mefenamic acid, and 4-biphenylacetic acid.

[0066] In another embodiment, a compound of formula III or its pharmaceutically acceptable salt, ester, hydrate, chelate, and / or solvate is provided:

[0067]

[0068] in As defined above; and NSAID is the remainder, which, together with the carboxyl group to which it is attached, forms a nonsteroidal anti-inflammatory drug (NSAID) compound.

[0069] In one embodiment, the compound comprising the isotope-enriched amide is rapatinel (GLYX-13, an NMDA receptor modulator), safinamide (a monoamine oxidase inhibitor), and / or BPN14770 (2-(4-((2-(3-chlorophenyl)-6-(trifluoromethyl)pyrimidin-4-yl)amino)phenyl)acetamide, a PDE4 allosteric modulator). The corresponding non-isotope-enriched drug structures are given below:

[0070]

[0071] In another embodiment, the amide-containing drug comprises an isotopically enriched amide functional group, wherein one or more of the O, N, and C atoms in the amide functional group are isotopically enriched or labeled, and the amide-containing drug is one of the following:

[0072]

[0073] It should be understood that the re-stable isotope effect can also be applied to esters, for example, when the ester functional group becomes crucial during the generation of new chemical entities via ester hydrolysis, such as in the case of carboxylic acid esters (RCOOR') or phosphate esters (ROP(O)(OR')OR''). Therefore, ester-containing pharmaceutical and / or prodrug compounds are provided that have one or more re-stable isotopes of naturally occurring isotopes in one or more carboxylic acid or phosphate ester functional groups, replacing those of the pharmaceutical and / or prodrug compounds. In some embodiments, isotope-enriched esters are provided, wherein the ester is part of a pharmaceutical or prodrug.

[0074] In other embodiments, this technology can be applied to RNA interference (RNAi) therapeutic agents, such as, but not limited to, synthetic small interfering RNAs (siRNAs), including siRNA-drug and / or siRNA-prodrug conjugates and GalNAc-siRNA conjugates.

[0075] Biantennary and triantennary GalNAc ligands can be covalently conjugated to siRNA (see, for example, Khorev, O.; et al., Bioorg. Med. Chem. 2008, 16, 5216; Rensen, PCN; et al., J. Med. Chem. 2004, 47, 5798; Valentijn, ARPM; et al., Tetrahedron, 1997, 53, 759; Manoharan, M.; et al., WO2009073809, 2009). Some examples of the synthesis of such siRNA-GalNAc conjugates can be found, for example, Nair, JK et al., J. Am. Chem. Soc. 2014, 136, 16958-16961, as follows:

[0076]

[0077] Nair, JK, et al. designed a variety of GalNAc-siRNA conjugates (Nucleic Acids Research, 2017, 45(19), 10969–10977), which are shown in the table below:

[0078]

[0079] S and AS represent sense and antisense chains, respectively; uppercase and lowercase letters indicate 2'-deoxy-2'-fluoro(2'-F) and 2'-O-methyl(2'-OMe) ribosugar modifications, respectively; * indicates PS bonding. L indicates a trivalent GalNAc ligand (as described above).

[0080] It is generally believed that after uptake, GalNAc-siRNA fractions reach the cytoplasm and are loaded into the Argonaute protein within the RNA-induced silencing complex (RISC). Subsequently, the sense (or "lackey") strand of the siRNA is released, while the antisense (or "guide") strand facilitates sequence-specific enzymatic cleavage by guiding the RISC to complementary RNA, thereby reducing protein expression in a highly targeted manner (Elbashir, SM; et al., Nature, 2001, 411, 494–98). Therefore, GalNAc-siRNA conjugates with multiple functional groups or organic bonds (which are amides, esters, and / or ethers) between their GalNAc and siRNA components are expected to release siRNA into the cytoplasm by directly cleaving the relevant chemical bonds at phosphate sites or by initiating cleavage at one or more amide sites. In fact, as discussed above, targeted delivery of siRNA to the liver has been achieved using synthetic trivalent N-acetylgalactosamine (GalNAc) ligands covalently conjugated to chemically modified small interfering RNA (siRNA). This strategy enables safe and efficient targeted delivery of siRNA to the liver via the asialoglycoprotein receptor (ASGPR) located on the surface of hepatocytes, which triggers clathrin-mediated endocytosis, thereby enabling intracellular delivery of siRNA to induce RNAi-mediated RNA silencing (Nair, J.K. et al. Nucleic Acids Research, 2017, 45(19), 10969–10977). Such intracellular delivery of siRNA generally involves cleaving the covalent bond between the 3'-O of the sense strand and the linker moiety by directly cleaving the PO bond, or by triggering the cleavage of the amide bond in the spacer region, as shown here:

[0081]

[0082] In the examples above, the structure has at least two types of cleavable covalent bonds: amide bonds in the spacer region and phosphate bonds in the linker region, and can be cleaved at the indicated locations (dashed lines).

[0083] Therefore, in one embodiment, a GalNAc-siRNA conjugate is provided having one or more restable isotopes that replace naturally occurring isotopes in one or more amide groups of the conjugate. In some embodiments, one or both amide oxygen atoms in the GalNAc-siRNA conjugate are enriched with... 18O. Not wanting to be limited by theory, it is assumed that one or more amide oxygen atoms in the spacer region... 18 O enrichment can improve the rate of amide hydrolysis (chemically or enzymatically), which in turn can lead to a change in the rate of intracellular release of RNAi in cases where amide cleavage results in the final cleavage of the phosphate bond between the linker and the RNAi moiety. In other embodiments, one or more oxygen atoms in the phosphate ester bond of the linker of the GalNAc-siRNA conjugate are... 18 O may be partially or completely substituted. It is not desirable to be limited by theory, but rather to consider when the P atom is to the right (and / or left) of the P atom. 16 O was 18 When O is substituted, the bond energies of both PO and OC change, which in turn improves the hydrolysis rate at that site.

[0084] In GalNAc-siRNA conjugates according to some embodiments of this technology 18 Some non-restrictive examples of O-isotope enrichment are shown below, where the linker phosphate group is attached to RNAi at the 3'-position:

[0085]

[0086] In other implementations, the above examples are further extended to cover a variety of interval lengths, as follows:

[0087]

[0088] Where n is an integer from 7 to 25. It should be understood that the target site in the GalNAc-siRNA conjugate is enriched with […]. 18 O, or enriched with 18 O、 15 N and / or 13 Other combinations of stable isotopes of C are also considered and intended to be covered herein.

[0089] In one specific implementation, the isotope-enriched compounds listed in Table 1 are provided, as well as their pharmaceutically usable salts, esters, hydrates, chelates, and solvates.

[0090] Table 1. Some examples of isotope enrichment compounds using this technique.

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097] Many drugs and prodrugs have amide functional groups at key sites in their structures, and numerous examples of drugs and prodrugs containing isotopically enriched amides are available. It should be understood that the inclusion of isotopically enriched amide functional groups in drugs and prodrugs of this technology is not intended to impose any particular limitation.

[0098] As shown below, other types of amides and their pharmaceutically acceptable salts, esters, hydrates, chelates, and / or solvates are also within the scope of this invention. Therefore, in another embodiment, the following compounds and their pharmaceutically acceptable salts, esters, hydrates, chelates, and solvates are provided:

[0099]

[0100] Among them: O * It is enriched with stable heavy oxygen isotopes ( 17 O and / or 18 The amide oxygen atom of O); R, R 1 and R 2 As defined above; and isotopic enrichment structures exist in O containing at least one amide bond. * The enriched drug and / or prodrug compound, or the isotopically enriched structure, provides an O-containing element comprising at least one amide bond. * Enriched drug and / or prodrug compounds, wherein the amide linkage is, for example, but not limited to, phosphonamide linkage, phosphoramide linkage and / or sulfonamide linkage.

[0101] In a second, broader aspect, pharmaceutical compositions are provided comprising the isotope-enriched compounds described herein or their pharmaceutically acceptable salts, esters, hydrates, chelates, or solvates, and a pharmaceutically acceptable carrier.

[0102] In the third broad aspect, a method is provided for modulating or improving the pharmacokinetic profile of a drug or prodrug by replacing one or more of the three naturally occurring atoms (C(=O)-N) in the amide functional group or the bond with one or more restable isotopic atoms.

[0103] In one embodiment, compared with compounds using only atoms with natural isotopic abundance (i.e., non-isotopically enriched compounds), the isotopically enriched compounds described herein (e.g., compounds of formula I, II, or III, compounds in Table 1) and / or their pharmaceutical compositions are used in a subject to modulate the metabolic pathway of the drug or prodrug, reduce drug metabolism of the drug or prodrug, modulate the pharmacokinetic profile of the drug or prodrug, and / or improve or enhance the therapeutic effect of the drug or prodrug. In some embodiments, compared with compounds using only isotopes with natural abundance (i.e., non-isotopically enriched compounds), the isotopically enriched compounds and pharmaceutical compositions provided herein are used in a subject to reduce the therapeutic toxicity and / or adverse effects of the compound, improve the tolerability of the compound, and / or improve or enhance the therapeutic or prophylactic effect of the compound. In some embodiments, the isotope-enriched compounds and pharmaceutical compositions provided herein are used in subjects to improve the biodistribution of the compounds in the subjects and / or enhance the therapeutic and / or preventive effects of the compounds, compared to compounds using only isotopes with natural abundance (i.e., non-isotope-enriched compounds).

[0104] In one embodiment, a method is provided for modulating the metabolic or pharmacokinetic profile of an amide-containing (i.e., amide-functional group-containing) compound in a subject, comprising administering to the subject an isotopically enriched compound and / or pharmaceutical composition as described herein, wherein the metabolic or pharmacokinetic profile of the isotopically enriched amide-containing compound is modulated compared to administration of the same compound having only a naturally abundant isotope (i.e., a non-isotopically enriched compound). In some embodiments, a method is provided for reducing the metabolism of a compound, reducing the therapeutic toxicity of a compound, reducing the adverse effects of a compound, improving the tolerability of a compound, improving the biodistribution of a compound, and / or improving the therapeutic or prophylactic effects of a compound in a subject, said method comprising administering to the subject an isotopically enriched compound or pharmaceutical composition as described herein, wherein the isotopically enriched compound exhibits reduced metabolism, reduced therapeutic toxicity, reduced adverse effects, improved tolerability, improved biodistribution, and / or improved therapeutic or prophylactic effects compared to administration of a compound having only a naturally abundant isotope (i.e., a non-isotopically enriched compound).

[0105] In a fourth, broader aspect, methods are provided for treating disease states or conditions associated with the activity of phosphoinositide 3-kinase in subjects with this need, the methods comprising administering to the subject a therapeutically effective amount of an isotopically enriched compound and / or pharmaceutical composition as described herein. Any disease or condition that improves by inhibition of phosphoinositide 3-kinase (referred to herein as “PI3K-mediated disease”) can be treated using the compounds and compositions provided herein. In some embodiments, phosphoinositide 3-kinase (PI3K)-mediated disease is a PI3Kα-mediated disease (or a disease mediated by overexpression or amplification of PI3Kα, somatic mutations of PI3K, germline mutations or somatic mutations of PTEN, or mutations and translocations of p85α for upregulating the p85-p110 complex), particularly such a condition responding in a beneficial manner to inhibition of PI3Kα, particularly to inhibition of PI3Kα or its mutant forms.

[0106] In some implementations, the disease state or condition to be treated is a proliferative disorder, such as PI3K-mediated tumor and / or cancer cell growth. The disease may include those exhibiting overexpression or amplification of PI3Kα, somatic mutations in PIK3CA, germline or somatic mutations in PTEN, or mutations and translocations of p85α that upregulate the p85-p110 complex. Specifically, the compounds can be used to treat cancers in humans or animals (e.g., mice), including, for example: sarcoma; lung; bronchus; prostate; breast (including sporadic breast cancer and Cowden disease); pancreas; gastrointestinal cancer; colon; rectum; colon cancer; colorectal adenoma; thyroid; liver; intrahepatic bile ducts; hepatocytes; adrenal glands; stomach; gastric region; glioma; glioblastoma; endometrium; melanoma; kidney; renal pelvis; bladder; uterine body; cervix; vagina; ovary; multiple myeloma; esophagus; leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; brain; brain cancer; oral cavity and pharynx; larynx; small intestine; non-Hodgkin lymphoma; melanoma; villous colonic adenoma; tumor formation; epithelial-characteristic tumor formation; lymphoma; breast cancer (mammary tumor). (carcinoma); basal cell carcinoma; squamous cell carcinoma; actinic keratosis; neoplastic diseases, including solid tumors; tumors of the neck or head; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; and Walden stroem's disease. In one embodiment, the disease state or condition to be treated is breast cancer. In some such embodiments, the disease state or condition to be treated is hormone receptor positive (HR). +), human epidermal growth factor receptor negative (HER2) - Breast cancer. In another implementation, the disease state or condition to be treated is HR. + HER2 - And breast cancer with PIK3CA mutation.

[0107] In other implementations, PI3K-mediated conditions or disorders are selected from: polycythemia vera, essential thrombocythemia, myelofibrosis with myeloid metaplasia, asthma, COPD, ARDS, Loffler syndrome, eosinophilic pneumonia, parasitic (especially metazoan) infections (including tropical eosinophilia), bronchopulmonary aspergillosis, polyarteritis nodosa (including Churg-Strauss syndrome), eosinophilic granuloma, eosinophilic-associated conditions affecting the airways due to drug reactions, psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, herpetic dermatitis. Dermatitis, scleroderma, vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphigus, acquired epidermolysis bullosa, autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell anemia, and idiopathic thrombocytopenic purpura), systemic lupus erythematosus, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic stomatitis. This includes conditions such as sprue, autoimmune inflammatory bowel diseases (e.g., ulcerative colitis and Crohn's disease), endocrine eye diseases, Graves' disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonia, multiple sclerosis, primary biliary cirrhosis, uveitis (pre- and post-), ​​interstitial pulmonary fibrosis, psoriatic arthritis, glomerulonephritis, cardiovascular diseases, atherosclerosis, hypertension, deep vein thrombosis, stroke, myocardial infarction, unstable angina, thromboembolism, pulmonary embolism, thrombolytic disease, acute arterial ischemia, peripheral thrombotic occlusion, and coronary artery disease, reperfusion injury, retinopathy (e.g., diabetic retinopathy or hyperbaric oxygen-induced retinopathy), and conditions characterized by aqueous humor secretion or increased intraocular pressure (e.g., glaucoma).

[0108] In some embodiments, the disease state or condition is a solid tumor or cancer or proliferative disease, such as, but not limited to: breast cancer, melanoma, colon cancer, colorectal adenoma, pancreatic cancer, prostate cancer, lung cancer, leukemia, brain tumor, lymphoma, ovarian cancer, Kaposi's sarcoma, rectal cancer, renal cell carcinoma, glioblastoma, non-small cell lung cancer, head and neck cancer, multiple myeloma, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, or uveal melanoma. Therefore, in some embodiments, a method of treating cancer, solid tumor, or proliferative disease is provided, comprising administering to a subject a therapeutically effective amount of an isotope-enriched compound and / or pharmaceutical composition as described herein, thereby treating the cancer, solid tumor, or proliferative disease.

[0109] In another broad aspect, kits are provided that contain one or more isotopically enriched compounds or pharmaceutical compositions described herein. The kit may also contain one or more additional therapeutic agents and / or instructions, for example, instructions for using the kit to treat a subject suffering from the same or similar disease state or condition as that treated by the parent (i.e., non-isotopically enriched) compound. Attached Figure Description

[0110] To better understand the invention and to more clearly show how it can be implemented, reference will now be made to the accompanying drawings by way of example, which illustrate aspects and features of some embodiments of the invention, and in which:

[0111] Figure 1a and 1b The figures show the results of oral administration of the same doses of apelips and apelips- 18 apelips (dashed line) and apelips- in O1 SD rats 18 Plasma drug concentration-time curves at O1 (solid line): (1a) at a dose of 2.5 mg / kg and (1b) at a dose of 0.322 mg / kg.

[0112] Figure 2 The figures show the results of oral administration of the same doses of apelips and apelips- 18 Apellipril (-Δ-) and Apellipril- in O1 ICR mice 18 Plasma drug concentration-time curve for O1(-○-).

[0113] Figure 3a , 3b Figures 3c and 3d show the incubation of apelips and apelips- in a medium containing liver S9 from multiple species. 18 After O1, M4 and M4- 18O1 production: (3a) monkey liver S9, (3b) combined human S9, (3c) mini pig liver S9, and (3d) male Wistar rat liver S9. In each figure: -x-, M4; -o-, M4- 18 O.

[0114] Figure 4 The same dose of apelips and apelips- were shown when administered orally. 18 Apellis (-△-) and Apellis- in O1 Beagle 18 Plasma drug concentration-time curve for O1(-○-).

[0115] Figure 5 It shows 18 An example of the chemical synthesis pathway for O1-apelis. Detailed Implementation

[0116] definition

[0117] To provide a clear and consistent understanding of the terminology used in this specification, numerous definitions are provided below. Furthermore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0118] When used in conjunction with the term "comprising / including" in the claims and / or description, the use of a noun without a quantifier may mean "one / type," but it also carries the meaning of "one / type or more / types," "at least one / type," and "one / type or more than one / type." Similarly, the word "another" may indicate at least a second or more.

[0119] As used in this article, the term "naturally abundant atom" refers to an atom with a natural isotopic abundance, typically having an isotopic composition close to that of atoms in Earth's atmosphere. However, it should be understood that naturally occurring compounds can exhibit slight variations in the isotopic composition of many atoms. For example, the term "naturally abundant oxygen" refers to oxygen atoms with a natural isotopic abundance, typically having an isotopic composition close to that of oxygen atoms in Earth's atmosphere. 16 O, 99.759%; 17 O, 0.037%; and 18 O, 0.204%. However, it should be understood that naturally occurring compounds can have slight variations in the isotopic composition of oxygen atoms.

[0120] Terms “heavy oxygen atom”, “heavy oxygen isotope”, “stable heavy oxygen” * O" and "O * "Can be used interchangeably to refer to" 17 O and / or18 O is a stable oxygen atom, excluding any radioactive or non-naturally occurring heavy isotopes. 17 O and 18 O is naturally occurring, but it is different from the main isotopes. 16 O exists in a very low proportion compared to it. Terms such as "heavy carbon isotope," "stable heavy carbon isotope," and "carbon-13" are also mentioned. 13 C and " * "C" can be used interchangeably in this article to refer to... 13 C isotopes. Similarly, the terms "diazonium isotope," "stable diazonium isotope," "nitrogen-15," and "[other related terms]" are used. 15 N and “ * "N" can be used interchangeably in this article to refer to... 15 Nitrogen isotopes.

[0121] The terms used in this article, such as "oxygen atoms enriched with stable dioxane isotopes", "stable dioxane enrichment", and "dioxane enrichment", are: * "O-enrichment" and "O" * "Enrichment" is used interchangeably to refer to oxygen atoms that are not present in the natural isotopic composition and have a higher level of stable heavy oxygen isotopes than those present in the naturally occurring isotopic composition. The terms "heavy oxygen enrichment" and "stable heavy oxygen enrichment" are also interchangeable. * "O-enrichment" and "O" * "Enrichment" can be used interchangeably, referring to the enrichment of stable tartrate isotopes, as described in this article, at specific sites on a compound. 17 O, or 18 O, or 17 O and 18 Oxygen atoms in a mixture of O atoms. Similarly, it mentions enrichment in stable heavy atomic isotopes (respectively...). 13 C enrichment or 15 (N-enriched) carbon or nitrogen atoms refer to carbon or nitrogen atoms that are not present in the natural isotopic composition, including heavy isotopes in compounds. 13 C or 15 The fraction of N is higher than that found in naturally occurring carbon or nitrogen.

[0122] Isotope enrichment is a process in which the relative abundance of isotopes of a given element is altered, resulting in the enrichment (i.e., increase) of one particular isotope and the depletion or exhaustion of its other isotopic forms. As used herein, “heavy isotope-enriched” compounds or derivatives refer to compounds in which a particular isotopic form has been increased at a specific position or site of amide bonding; that is, oxygen-17 or oxygen-18, or both oxygen-17 and oxygen-18, and / or carbon-13, and / or nitrogen-15 have been enriched (i.e., increased).

[0123] For carbon and nitrogen, with the most abundant isotopes 12 C and 14 The most abundant heavy isotope is N, with levels of 0.9893 and 0.99636 on the Mohr fraction scale, respectively. 13 C and 15 N, its levels, expressed as molar fractions, are 0.0107 and 0.00364. Under normal conditions, oxygen-18 ( 18 O) and oxygen-17 ( 17 The levels of O) were 0.00204 and 0.00037, respectively, and the most abundant oxygen isotope was oxygen-16 ( 16 The level of O) was 0.99757, all expressed as molar fractions. Therefore, 18 O and 16 The ratio of O is approximately 0.2%.

[0124] The "isotope-enriched" compounds or derivatives used in this article have a higher abundance than the natural abundance forms of atoms or elements (e.g., 16 O) at higher levels of atoms or isotopic forms of elements (e.g., 18 The level of isotopic enrichment will vary depending on the natural abundance of a particular isotopic form. As used herein, the expression "level of isotopic enrichment" refers to the amount or percentage of a compound containing an isotopic form of an atom or element, replacing the isotope with the highest natural abundance. The terms "level of enrichment" and "enrichment %" are used interchangeably and refer to the amount or molar percentage of a compound containing an isotopic form of an atom or element. For example, "95%" 18 "O enrichment" means that 95 out of 100 molecules have [a specific characteristic / enrichment]. 18 O isotope, and 5 / 100 do not have 18 O, but rather oxygen in other isotopic forms ( 17 O and / or 16 O). Similarly, at a certain position / atom within the compound structure, 95%. 18 O-enrichment means that 95 / 100 molecules at this position / atom in the compound have [a certain concentration / absorption rate]. 18 O, while the other 5 / 100 have other isotopic forms of oxygen at the same position / atom ( 17 O and / or 16In some embodiments, the isotopic enrichment level of the compound or the isotopic enrichment level of the elements in the compound can be from about 2 to about 100 molar percentages (%), for example, about 2%, about 5%, about 17%, about 30%, about 51%, about 83%, about 90%, about 95%, about 98%, greater than about 98%, about 99%, or 100%. In one embodiment, the isotopic enrichment level in the isotopic enriched compound of the present invention (e.g., compounds of any of Formulas I to III, or compounds in Table 1, etc.) is about 5% or higher, or about 10% or higher. In another embodiment, the isotopic enrichment level in the isotopic enriched compound of the present invention (e.g., compounds of any of Formulas I to III, and compounds described herein) is about 20% or higher, or about 50% or higher. In yet another embodiment, the isotope enrichment level of the isotope-enriched compound of the present invention (e.g., compounds of any of Formulas I to III, and compounds described herein) is about 75% or higher, or about 90% or higher. In yet another embodiment, the isotope enrichment level of the isotope-enriched compound of the present invention (e.g., compounds of any of Formulas I to III, and compounds described herein) is about 90% or higher, about 91% or higher, about 92% or higher, about 93% or higher, about 94% or higher, about 95% or higher, about 96% or higher, about 97% or higher, about 98% or higher, about 99% or higher, or 100%. In some embodiments, the isotope enrichment level of the compound is determined using mass spectrometry. It should be understood that the isotopic enrichment level of a particular compound or the isotopic enrichment level of a particular oxygen isotope of the compound will be selected based on several properties of the compound (e.g., its chemistry, pharmacokinetics, and therapeutic profile), with the aim of improving the compound's therapeutic or prophylactic efficacy, therapeutic biodistribution, bioavailability, metabolism, stability, and / or pharmacokinetic profile, and / or reducing the compound's adverse effects.

[0125] The “non-isotope-enriched” compounds used herein are those in which all atoms or elements are naturally abundant isotopes, meaning all atoms or elements have the most abundant atomic masses found in nature. This contrasts with isotope-enriched compounds, where one or more elements are enriched in forms that are not naturally abundant isotopes. Non-isotope-enriched compounds are excluded from the compounds of this invention provided herein.

[0126] As used herein, the terms “compound of the present invention”, “compound of the present invention”, and equivalent expressions refer to isotope-enriching compounds provided herein that may be used for at least one of the purposes described herein, such as those covered by structural formulas (e.g., Formulas I to III) and include the specific compounds described herein, as well as their pharmaceutically acceptable salts, esters, chelates, hydrates, and / or solvates.

[0127] The "isotope effect" used in this article refers to the influence of stable heavy isotopes (which replace naturally abundant isotopes) on the amide bond cleavage rate. Without being bound by theory, heavy isotopes can alter the strength of the amide bond due to their increased mass, and thus cause a decrease in the amide bond cleavage reaction rate. For amide hydrolysis, the key rate-determining step is typically the attack of water molecules on the carbon atoms in the planar amide group. As the oxygen atom in the water molecule approaches, the π bond in the initial carbon-oxygen double bond gradually breaks, thus forming a four-coordinate intermediate. If the strength or rigidity of the carbon-oxygen double bond is altered, the hydrolysis rate can be changed accordingly. Therefore, the introduction of heavy stable isotopes can slow down the hydrolysis of amides to carboxylic acids. This change in the amide bond cleavage rate can improve the pharmacokinetic profiles of drugs and prodrugs containing amide bonds, and thus modulate the therapeutic, prophylactic, and / or adverse effects of drugs and prodrugs. Since the carboxylamide center involves three atoms, oxygen, carbon, and nitrogen, stable heavy isotopes of oxygen and / or carbon and / or nitrogen may potentially have isotopic effects on drugs and prodrugs containing amide bonds.

[0128] Unrestricted by theory, the two deuterium isotopes, namely 18 O and 17 O can have different isotopic effects with respect to the rate of amide bond cleavage (e.g., hydrolysis). Due to 18 The atomic weight of oxygen is 18 Daltons, therefore in some cases, due to its high mass, it is similar to... 17 Compared to O (17-Dalton atomic weight), it can exhibit a higher isotopic effect. (Optional) 18 O or 17 O, or multiple proportions 18 O and 17 Enrichment of mixtures of O can be achieved to realize desired biological and pharmaceutical effects. Similarly, heavy isotopes can be selected. 13 C and / or 15 Nitrogen enrichment is used to achieve the desired biological and pharmaceutical effects of isotopically enriched compounds. 17 Similarly, in some cases... 13 C and 18 O exhibits a lower isotopic effect compared to its native abundance isotope, due to its smaller molecular weight variation (molecular weight increment from 0.05%). 12 C to 13 C is 8.3%; from 16 O to 17 O was 6.25%; and from 16 O to 18 O was 12.5%.

[0129] As described in this article, isotope enrichment compounds can be made using one or more isotopes of the same element or using isotopes of one or more elements.

[0130] As used herein, "drug," "prodrug," or "parent compound" refers to a compound having a natural isotopic abundance at the target amide site. This technology provides compounds, compositions, and methods for preparing and using such compounds in a "heavy isotope-enriched" form to modulate or improve the pharmacological profile and / or therapeutic efficacy of the parent compound. In some embodiments, "drug," "prodrug," or "parent compound" refers to a compound having a natural oxygen, carbon, and / or nitrogen isotopic abundance at the respective target amide-oxygen, carbon, and / or nitrogen sites; therefore, this technology provides compounds, compositions, and methods for preparing and using such compounds in a "heavy oxygen, carbon, and / or nitrogen-enriched" form to modulate or improve the pharmacological profile and / or therapeutic efficacy of the parent compound.

[0131] As used in this article, "organic moiety" or "organic segment" refers to a group of atoms that form part of the overall structure of an isotope-enriched compound. The organic moiety, amide bond or functional group, and protecting or substituent (if present) are usually linked together by covalent bonds to form the structure of the isotope-enriched compound.

[0132] In one embodiment, the isotope-enriched compound may also be substituted with one or more substituents, when such substitution is available. In some embodiments, the substituted form of the compound is a prodrug; in such embodiments, the substituents may be cleaved, or the compound may be otherwise converted to release the active ingredient or pharmaceutical compound from the prodrug form after administration to a target.

[0133] As used herein, the term “application” or variations thereof refers to the delivery of a compound to a subject, including all methods of administration and drug delivery known in the art.

[0134] The terms “comprising” (and any variations thereof), “having” (and any variations thereof), “including” (and any variations thereof), and “containing” (and any variations thereof) as used herein are inclusive or open-ended and do not exclude additional elements or process steps not listed.

[0135] The term “about / approximately” as used herein is used to indicate that the value includes the inherent variation of error in the means or method used to determine the value.

[0136] The term “derivative” as used in this article refers to a substance that is structurally similar to another compound but differs in some minor structural details.

[0137] This specification involves many chemical terms and abbreviations used by those skilled in the art. However, for clarity and consistency, definitions of the selected terms are provided.

[0138] As used herein, the terms “substituted” or “having substitution” refer to a compound or part having at least one (1) substituent. The terms “unsubstituted” or “not having substitution” refer to a compound or part having no other substituents except for an undetermined valence chemically saturated with hydrogen atoms.

[0139] As used herein, “substituent” or “substituent group” means a group selected from the following: halogen (F, Cl, Br or I), hydroxyl, mercapto, amino, nitro, carbonyl, carboxyl, alkyl, alkoxy, alkylamino, aryl, aryloxy, arylamino, acyl, thionyl, sulfonyl, phosphonyl, and other organic moieties used and accepted in general organic chemistry.

[0140] As used herein, the term "alkyl" refers to a saturated hydrocarbon having 1 to 12 carbon atoms, including linear, branched, and cyclic alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, isopropyl, tert-butyl, sec-butyl, isobutyl, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The term alkyl includes both unsubstituted and substituted alkyl groups. The term "C1-C" is also used. n "alkyl", where n is an integer from 2 to 12, refers to an alkyl group having 1 to the number of carbon atoms indicated by "n". Alkyl residues may be substituted or unsubstituted. In some embodiments, for example, the alkyl group may be substituted with a hydroxyl, amino, carboxyl, carboxylic acid ester, amide, carbamate, or aminoalkyl group.

[0141] Unless otherwise specified, the term “lower aliphatic,” “lower alkyl,” “lower alkenyl,” and “lower alkynyl” as used herein means that the moiety has at least one (two for alkenyl and alkynyl) and equal to or less than six carbon atoms.

[0142] The terms "cycloalkyl," "alicyclic," "carbocyclic," and their equivalents refer to groups that contain saturated or partially unsaturated carbocyclic rings in monocyclic, spirocyclic (sharing a single atom), or fused (sharing at least one bond) carbocyclic systems having three to fifteen ring members. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopenten-1-yl, cyclopenten-2-yl, cyclopenten-3-yl, cyclohexyl, cyclohexen-1-yl, cyclohexen-2-yl, cyclohexen-3-yl, cycloheptyl, bicyclo[4,3,0]nonyl, norbornyl, etc. The term cycloalkyl includes both unsubstituted and substituted cycloalkyl groups.n "Cycloalkyl", where n is an integer from 4 to 15, refers to a cycloalkyl group having 3 to the number of carbon atoms indicated by "n" in its ring structure. Unless otherwise specified, "lower cycloalkyl" as used herein has at least 3 and equal to or less than 8 carbon atoms in its ring structure.

[0143] Cycloalkyl residues can be saturated or contain one or more double bonds within the ring system. Specifically, they can be saturated or contain one double bond within the ring system. In unsaturated cycloalkyl residues, the double bond can be present in any suitable position. Monocycloalkyl residues are, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, or cyclotetradecyl, and they can also be, for example, C10, cyclohexyl, cyclohexenyl, cyclohexylene, cyclohexylene, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, or cyclotetradecyl, and they can also be, for example, C10, cyclohexylene ... 1-4 Alkyl substitution. Some examples of substituted cycloalkyl residues are 4-methylcyclohexyl and 2,3-dimethylcyclopentyl. Some examples of bicyclic ring structures are norbornene, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and bicyclo[3.2.1]octane.

[0144] The term "heterocyclic alkyl" and its equivalents refer to a group containing a saturated or partially unsaturated carbocyclic ring in a mono- or spirocyclic (sharing a single atom) or fused (sharing at least one bond) carbocyclic ring system having three to fifteen ring members, containing one to six heteroatoms (e.g., N, O, S, P) or a group containing such heteroatoms (e.g., NH, NR). x (R x (These can be alkyl, acyl, aryl, heteroaryl, or cycloalkyl groups), PO2, SO, SO2, etc. Where possible, heterocyclic alkyl groups can be C-linked or heteroatom-linked (e.g., via a nitrogen atom). Some examples of heterocyclic alkyl groups include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, tetrahydrodithienyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazine, aza-butane, oxepanyl, thioheptanyl, homopiperidinyl, oxepanyl, thiepanyl, and oxadiazine. oxazepinyl, diazepine diazepinyl, thio-azo The term heterocyclic alkyl includes thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dialkyl, 1,3-dioxolanecycloalkyl, pyrazolinyl, dithialanyl, dithiolanyl, dihydropyranyl, dihydrothiophenyl, dihydrofuranyl, pyrazolyl, imidazolinyl, imidazolinyl, 3-azabicyclo[3,1,0]hexyl, 3-azabicyclo[4,1,0]heptyl, 3H-indolyl, quinazinyl, and sugars, etc. The term heterocyclic alkyl includes both unsubstituted and substituted heterocyclic alkyl groups. The term "C3-C" is used in conjunction with the preceding text. n "Heterocyclic alkyl", where n is an integer from 4 to 15, refers to a heterocyclic alkyl group having 3 to the number of atoms indicated by "n" in its ring structure, containing at least one heterogroup or atom as defined above. Unless the number of carbons is otherwise specified, "lower heterocyclic alkyl" as used herein refers to a heterocyclic alkyl group having at least 3 and equal to or less than 8 carbon atoms in its ring structure.

[0145] The terms "aryl" and "aromatic ring" refer to an aromatic group having "4n+2" π (pi) electrons and six to fourteen ring atoms in a conjugated monocyclic or polycyclic system (fused or unfused), where n is an integer from 1 to 3. Polycyclic systems contain at least one aromatic ring. Aryl groups can be directly linked or linked via C1-C3 alkyl groups (also called arylalkyl or aralkyl groups). Some examples of aryl groups include, but are not limited to, phenyl, benzyl, ethoxyphenyl, 1-phenylethyl, tolyl, naphthyl, biphenyl, terphenyl, indene, benzocyclooctenyl, benzocycloheptenyl, azulel, acenaphthel, fluorenyl, phenanthrene, anthracene, etc. The term aryl includes both unsubstituted and substituted aryl groups. The term "C6-C" is also used. n "Aryl", where n is an integer from 6 to 15, refers to an aryl group having 6 to the number of atoms shown "n" in a ring structure, which contains at least one heterogroup or atom as defined above.

[0146] The terms "heteroaryl" and "heteroaryl ring" refer to an aromatic group having "4n+2"π(pi) electrons and five to fourteen ring members in a conjugated monocyclic or polycyclic system (fused or unfused), where n is an integer from 1 to 3, and containing one to six heteroatoms (e.g., N, O, S) or groups containing such heteroatoms (e.g., NH, NR). x (R xIt can be alkyl, acyl, aryl, heteroaryl, or cycloalkyl (SO, etc.). Polycyclic systems contain at least one heteroaryl ring. Heteroaryl groups can be directly linked or linked via C1-C3 alkyl groups (also called heteroarylalkyl or heteroarylalkyl). Where possible, heteroaryl groups can be C-linked or heteroatom-linked (e.g., via a nitrogen atom). Some examples of heteroaryl groups include, but are not limited to, pyridinyl, imidazole, pyrimidinyl, pyrazolyl, triazolyl, tetrazolyl, furanyl, thiophene, and isoaryl. azole group, thiazolyl group, Azolyl, isothiazolyl, pyrrollyl, quinolinyl, isoquinolinyl, indole, isoindole, chromenyl, isochromenyl, benzimidazolyl, benzofuranyl, cinolinyl, indazole, indazinyl, phthalazinyl, pyridazinyl, pyrazinyl, triazinyl, isoindoleyl, pteridylyl, purineyl Diazolyl, thiadiazolyl, furazonyl, benzofurazanyl, benzothiophenyl, benzothienyl, benzothiazolyl, benzo[] Azolyl, quinazolinyl, quinazinyl, quinoloneyl, isoquinoloneyl, quinoxalinyl, naphthidyl, furanopyridyl, carbazolyl, phenanthridine, acridineyl, perimidinyl, phenanthrolinel, phenazinyl, phenthiazinyl, phenanthridine Zinel, dibenzofuranyl, etc. The term heteroaryl includes both unsubstituted and substituted heteroaryl groups. The term "C5-C" is also used. n "Heteroaryl", where n is an integer from 6 to 15, refers to a heteroaryl group having 5 to the number of atoms shown "n" in a ring structure, which contains at least one heteroaryl group or atom as defined above.

[0147] The term "heterocyclic" or "heterocyclic" includes heterocyclic alkyl and heteroaryl groups. Some examples of heterocycles include, but are not limited to, acridine, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophene, and benzo[…]. Azolyl, benzothiazolyl, benzotriazolyl, benzotetrazole, benzoisocyanate Azolyl, benzoisothiazolyl, benzoimidazolinyl, carbazole, 4αH-carbazole, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofurano[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolyl, imidazolinyl, imidazolyl, 1H-indolenyl, indolenyl, dihydroindoleyl, indazinyl, indoleyl, 3H-indoleyl, isobenzofuranyl, isochonyl, isoindoleyl, isodihydroindoleyl, isoindoleyl, isoquinolinyl, isothiazolyl, iso... Azolyl, methylenedioxyphenyl, morpholinyl, naphthidyl, octahydroisoquinolinyl Diazole group, 1,2,3- Diazolyl, 1,2,4- Diazolyl, 1,2,5- Diazolyl, 1,3,4- diazole group, azolealkyl, azole group, azolealkyl, pyrimidinyl, phenanthridine, phenanthroline, phenazinyl, phenthiazinyl, phenoxathiinyl, phen Azinyl, phthalazinyl, piperazinyl, piperidinyl, piperidinoneyl, 4-piperidinoneyl, piperinyl, pteridinyl, purine, pyranyl, pyrazinyl, pyrazolylyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridinyl Zazole, pyridinyl pyridoxazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolylyl group, pyrrolinyl group, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinazinyl, quinoxalinyl, quininecycloyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thiaanthryl, thiazolyl, thiophenyl, thiophenothiazolyl, thiophenothiazolyl Azolium, thiopheneimidazole, thiophenyl, triazine, 1,2,3-triazolium, 1,2,4-triazolium, 1,2,5-triazolium, 1,3,4-triazolium, xanthonium, etc. The term heterocyclic includes both unsubstituted and substituted heterocyclic groups.

[0148] The term "amine" or "amino" as used in this article refers to formula -NR a R b The unsubstituted or substituted portion, wherein R a and R b Each is independently hydrogen, alkyl, aryl, or heterocyclic, or R a and R bTogether with the nitrogen atoms to which they are attached, they form heterocyclic rings. The term amino includes compounds or portions in which the nitrogen atom is covalently bonded to at least one carbon or heteroatom. Thus, as used herein, the terms “alkylamino” and “dialkylamino” mean an amino group having one and at least two C1-C6 alkyl groups attached thereto, respectively. The terms “arylamino” and “diarylamino” include groups in which nitrogen is attached to at least one or two aryl groups, respectively. The terms “amide” or “aminocarbonyl” include compounds or portions containing a nitrogen atom attached to a carbon atom of a carbonyl or thiocarbonyl group. The term “acylamino” refers to an amino group directly attached to an acyl group as defined herein.

[0149] The term "alkyl thio" refers to an alkyl group having a thiol group attached thereto. Suitable alkyl thio groups include groups having 1 to about 12 carbon atoms, preferably 1 to about 6 carbon atoms. As used herein, the term "alkyl carboxyl" means an alkyl group having a carboxyl group attached thereto.

[0150] As used herein, the term "alkoxy" or "lower alkoxy" refers to an alkyl group having an oxygen atom attached to it. Representative alkoxy groups include those having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, tert-butoxy, etc. Some examples of alkoxy groups include methoxy, ethoxy, isopropoxy, propoxy, butoxy, pentoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy, dichloromethoxy, trichloromethoxy, etc. The term "alkoxy" includes both unsubstituted and substituted alkoxy groups, as well as perhalogenated alkoxy groups.

[0151] The terms "carbonyl" or "carboxyl" encompass compounds and moieties that contain a carbon atom linked to an oxygen atom by a double bond. Some examples of moieties containing a carbonyl group include aldehydes, ketones, carboxylic acids, amides, esters, acid anhydrides, etc.

[0152] The term "acyl" refers to a carbonyl group that is linked by a carbon atom to hydrogen (i.e., formyl), an aliphatic group (C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, such as acetyl), a cycloalkyl group (C3-C8 cycloalkyl), a heterocyclic group (C3-C8 heterocyclic alkyl and C5-C6 heteroaryl), or an aromatic group (C6 aryl, such as benzoyl). The acyl group can be unsubstituted or substituted (e.g., salicylyl).

[0153] The term "solvent" refers to the physical association of a compound with one or more solvents, whether organic or inorganic. This physical association includes hydrogen bonding. In some cases, solvates can be separated, such as when one or more solvents are incorporated into the crystal lattice of a crystalline solid. "Solvent" encompasses both the solution phase and the separable solvate. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and semi-ethanolates.

[0154] The term "medicinal salt" refers to a salt of a compound that is medicinally usable. Salts of compounds are desired to retain or improve the biological efficacy and properties of the free acids and bases of the same compound as defined herein, or to utilize the inherent basicity, acidity, or charged functional groups in the compound, and not in a way that would be undesirable from a biological or other perspective. Some examples of medicinal salts are described, for example, in Berge et al., "Pharmaceutical Salts", J. Pharm. Sci. 66, 1-19 (1977). Some non-limiting examples of such salts include:

[0155] (1) Acid addition salts are formed by adding the following inorganic acids to a basic or positively charged functional group: for example, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, aminosulfonic acid, nitric acid, phosphoric acid, carbonate forming agents, etc.; or by forming with the following organic acids: for example, acetic acid, propionic acid, lactic acid, oxalic acid, glycolic acid, neopentanoic acid, tert-butylacetic acid, β-hydroxybutyric acid, valeric acid, hexanoic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, succinic acid, malic acid, maleic acid, etc. Horse acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, cyclohexylaminosulfonic acid, benzenesulfonic acid, sulfanilic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 3-phenylpropionic acid, laurylsulfonic acid, laurylsulfonic acid, oleic acid, palmitic acid, stearic acid, lauric acid, embonic acid, palmoic acid, pantothenic acid, lactobionic acid, alginic acid, galactobionic acid, galacturonic acid, gluconic acid, glucoheponic acid, glutamic acid, naphthic acid, hydroxynaphthoic acid, salicylic acid, ascorbic acid, stearic acid, mucoconic acid, etc.

[0156] (2) Base addition salts are formed when the acidic protons present in the starting compound are replaced by metal ions, including alkali metal ions (e.g., lithium, sodium, potassium), alkaline earth ions (e.g., magnesium, calcium, barium) or other metal ions (e.g., aluminum, zinc, iron); or when coordinated with organic bases such as ammonia, ethylamine, diethylamine, ethylenediamine, N,N'-dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucosamine, piperazine, chloroprocaine, procaine, choline, lysine, etc.

[0157] Pharmaceutically acceptable salts can be synthesized from starting compounds containing a basic or acidic moiety using conventional chemical methods. Generally, such salts are prepared by reacting a compound in its free acidic or basic form with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of both. Salts can be prepared in situ during the final separation or purification of the compound, or by reacting the compound in its free acidic or basic form with the desired corresponding base or acid alone and separating the resulting salt. The term "pharmaceutically acceptable salt" also includes zwitterionic compounds containing a cationic group covalently bonded to an anionic group, as these are "internal salts." It should be understood that all acidic, saltic, basic, and other ionic and nonionic forms of the compounds described herein are intended to be included. For example, if a compound is shown as an acid herein, its salt form is also included. Similarly, if a compound is shown as a salt, its acidic and / or basic forms are also included.

[0158] As used herein, the term "effective amount" refers to the amount or dose of a therapeutic agent, such as a compound, that provides the desired therapeutic, prophylactic, diagnostic, or prognostic effect in a subject after administration of a single or multiple doses. The effective amount can be readily determined by an attending or diagnosing physician using known techniques and by observing results obtained in similar situations. In determining the effective amount or dose of the administered compound, numerous factors are considered, including but not limited to: the subject's body size, age, and general health status; the specific disease involved; the extent or severity of the disease or condition to be treated; the individual subject's response; the specific compound administered; the route of administration; the bioavailability characteristics of the administered formulation; the chosen dosing regimen; the use of concomitant medications; and other relevant considerations.

[0159] "Pharmaceutical-grade" refers to drugs, pharmaceutical preparations, inert ingredients, etc., that are suitable for contact with the cells or tissues of humans and animals without excessive toxicity, incompatibility, instability, irritation, allergic reactions, etc., and whose benefits / risks are commensurate with a reasonable ratio. It typically refers to compounds or compositions approved or permitted by federal or state regulatory agencies, or listed in the United States Pharmacopeia or other recognized pharmacopoeias for use in animals and more specifically for human use.

[0160] "Pharmaceutical-grade carrier" refers to a diluent, excipient, excipient, carrier, or delivery vehicle that is administered together with a compound. The terms "pharmaceutical-grade carrier" and "pharmaceutical-grade delivery vehicle" are used interchangeably in this document.

[0161] "Pharmaceutical composition" means a composition comprising a compound as described herein and at least one component, said component comprising a pharmaceutically acceptable carrier, diluent, excipient, excipient, or loading agent, such as a preservative, filler, disintegrant, wetting agent, emulsifier, suspending agent, sweetener, flavoring agent, aromatizer, antibacterial agent, antifungal agent, lubricant, dispersant, etc., depending on the method of administration and the nature of the dosage form. "Prophylaxis" or "preventive" is intended to mean at least a reduction in the likelihood (or susceptibility) to a disease or condition (i.e., the absence of at least one clinical symptom leading to the disease in patients who are exposed to or susceptible to the disease but have not yet experienced or exhibited symptoms of the disease).

[0162] In some embodiments, “treatment” or variations thereof for any disease or condition means improving at least one disease or condition. In some embodiments, “treatment” or variations thereof means improving at least one bodily parameter, whether identifiable by the patient or not. In some embodiments, “treatment” or variations thereof means suppressing a disease or condition physically (e.g., stabilization of identifiable symptoms), physiologically (e.g., stabilization of bodily parameters), or both. In some embodiments, “treatment” or variations thereof means improving the quality of life of a subject in need or reducing symptoms or side effects of a disease. “Therapeutic effective amount” means the amount of a compound sufficient to achieve such treatment or prevention of a disease when administered to a subject for the purpose of treating or preventing the disease. “Therapeutic effective amount” will vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject suffering from the disease to be treated or prevented. As used herein, the term “therapeutic effective amount” means the amount of a compound or composition sufficient to prevent, treat, suppress, reduce, improve, or eliminate one or more causes, symptoms, or complications of a disease (e.g., cancer).

[0163] The term "object" includes animals, including mammals and humans, especially humans. Some non-limiting examples of objects include humans, monkeys, cattle, rabbits, sheep, goats, pigs, dogs, cats, rats, mice, and their transgenic species.

[0164] The terms “prodrug” and “equivalent expression” refer to agents that can be directly or indirectly converted into their active form in vitro or in vivo (see, for example, R.B. Silverman, 1992, “The Organic Chemistry of Drug Design and Drug Action,” Academic Press, Chapter 8; Bundgaard, Hans; Editor. Neth. (1985), “Design of Prodrugs,” p. 360; Elsevier, Amsterdam; Stella, V.; Borchardt, R.; Hageman, M.; Oliyai, R.; Maag, H.; Tilley, J. (ed.) (2007), “Prodrugs: Challenges and…”). Rewards, XVIII, 1470p. Springer. Prodrugs can be used to alter the biodistribution (e.g., preventing the drug from normally entering the reaction site of a protease) or pharmacokinetics of a particular agent. A wide variety of groups have been used to modify compounds to form prodrugs, such as esters, ethers, phosphates, etc. When a prodrug is administered to a subject, the group is cleaved by enzymatic or non-enzymatic, reduction, oxidation, hydrolysis, or other means to reveal the active form. As used herein, “prodrug” includes its pharmaceutically acceptable salts, pharmaceutically acceptable solvates, and crystalline forms of either. Although not strictly necessary, prodrugs are typically pharmacologically inactive until converted to their active form.

[0165] The term "ester" refers to a compound that can be represented by the formula RCOOR (carboxylic acid ester) or RSO3R' (sulfonate ester), usually formed by the reaction between an alcohol and a carboxylic acid or sulfonic acid, typically in the absence of water.

[0166] The term "amino acid" generally refers to an organic compound containing both a carboxylic acid group and an amino group. The term "amino acid" includes both "natural" and "non-natural" amino acids. Additionally, the term "amino acid" includes O-alkylated and N-alkylated amino acids, as well as amino acids with nitrogen- or oxygen-containing side chains (e.g., Lys, Cys, or Ser), wherein the nitrogen or oxygen atom has been acylated or alkylated. Amino acids can be pure L or D isomers or mixtures of L and D isomers, including (but not limited to) racemic mixtures.

[0167] The term "natural amino acid" and its equivalents refer to L-amino acids that are normally present in naturally occurring proteins. Some examples of natural amino acids include, but are not limited to, alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), β-alanine (β-Ala), and gamma-aminobutyric acid (GABA).

[0168] The term “non-natural amino acid” refers to any derivative of a natural amino acid, including D-type, as well as α- and β-amino acid derivatives. The terms “non-natural amino acid” and “non-natural amino acid” are used interchangeably herein. It should be noted that some amino acids classified as non-natural amino acids herein (e.g., hydroxyproline) may exist naturally in an organism or specific protein. Amino acids with a variety of different protecting groups suitable for immediate use in solid-phase peptide synthesis are commercially available. In addition to the twenty most common naturally occurring amino acids, some examples of the following non-natural amino acids and amino acid derivatives (common abbreviations in parentheses) may be used: 2-aminohexanoic acid (Aad), 3-aminohexanoic acid (β-Aad), 2-aminobutyric acid (2-Abu), α,β-dehydro-2-aminobutyric acid (8-AU), 1-aminocyclopropane-1-carboxylic acid (ACPC), aminoisobutyric acid (Aib), 3-aminoisobutyric acid (β-Aib), 2-amino-thiazoline-4- Carboxylic acids, 5-aminovaleric acid (5-Ava), 6-aminohexanoic acid (6-Ahx), 2-aminoheptanoic acid (Ahe), 8-aminooctanoic acid (8-Aoc), 11-aminoundecanoic acid (11-Aun), 12-aminododecanoic acid (12-Ado), 2-aminobenzoic acid (2-Abz), 3-aminobenzoic acid (3-Abz), 4-aminobenzoic acid (4-Abz), 4-amino-3-hydroxy-6-methylheptanoic acid (statin, Sta), amino Oxyacetic acid (Aoa), 2-aminotetrahydronaphthalene-2-carboxylic acid (ATC), 4-amino-5-cyclohexyl-3-hydroxyvalerate (ACHPA), p-aminophenylalanine (4-NH2-Phe), 2-aminopimelic acid (Apm), biphenylalanine (Bip), p-bromophenylalanine (4-Br-Phe), o-chlorophenylalanine (2-Cl-Phe), m-chlorophenylalanine (3-Cl-Phe), p-chlorophenylalanine (4-Cl-Phe), m-chlorotyrosine (3- Cl-Tyr), p-benzoylphenylalanine (Bpa), tert-butylglycine (TLG), cyclohexylalanine (Cha), cyclohexylglycine (Chg), desmodium (Des), 2,2-diaminopimelic acid (Dpm), 2,3-diaminopropionic acid (Dpr), 2,4-diaminobutyric acid (Dbu), 3,4-dichlorophenylalanine (3,4-Cl-2-Phe), 3,4-difluorophenylalanine (3,4-F2-Phe), 3,5-diiodotyrosine (3,5-I2-Tyr), N-ethylglycine (EtGly), N-ethylasparagine (EtAsn), o-fluorophenylalanine (2-F-Phe), m-fluorophenylalanine (3-F-Phe), p-fluorophenylalanine (4-F-Phe), m-fluorotyrosine (3-F-Tyr), homoserine (Hse), homophenylalanine (Hfe), homotyrosine (Htyr), hydroxylysine (Hyl), allohydroxylysine (aHyl), 5-hydroxytryptophan (5-OH-Trp), 3- or 4-hydroxyproline (3- or 4-Hyp), p-iodophenylalanine (4-I-Phe), 3-iodotyrosine (3-I-Tyr), dihydroindole-2-carboxylic acid (Idc), isodexin (Ide), alloisoleucine ( α-Ile), isoperidinic acid (Inp), N-methyl isoleucine (Melle), N-methyl lysine (MeLys), m-methyl tyrosine (3-Me-Tyr), N-methyl valine (MeVal), 1-naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal), p-nitrophenylalanine (4-NO2-Phe), 3-nitrotyrosine (3-NO2-Tyr), ortholeucine (Nle), orthovaline (Nva), ornithine (Orn), phosphotyrosine (H2PO3-Tyr), octahydroindole-2-carboxylic acid (Oic), penicillamine (Pen), pentafluorophenylalanine (F5-Phe), phenylglycine (Phg), piperidine acid (pipecolic acid) The following amino acids are listed: propargyl glycine (Pip), pyroglutamic acid (PGLU), sarcosine (Sar), tetrahydroisoquinoline-3-carboxylic acid (Tic), thiophene alanine, and thioproline (Th).

[0169] For the isotope-enriched compounds provided herein, in some embodiments, it is intended to also cover their salts, including pharmaceutically acceptable salts. Those skilled in the art will understand that many salt forms (e.g., TFA salts, tetrazolium salts, sodium salts, potassium salts, etc.) are possible; a suitable salt is selected based on considerations known in the art. The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic acid or base, including inorganic acids and inorganic bases, as well as organic acids and organic bases. For example, for compounds containing basic nitrogen, the salt may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids. Suitable pharmaceutically acceptable acid addition salts of the compounds provided herein include, but are not limited to, acetates, benzenesulfonic acid salts, benzoates, camphorsulfonates, citrates, ethanesulfonates, fumarates, gluconates, glutamates, hydrobromide salts, hydrochlorides, hydroxyethanesulfonates, lactates, maleates, malates, mandelates, methanesulfonates, mucilages, nitrates, pyruvates, pantothenates, phosphates, succinates, sulfates, tartaric acid, and p-toluenesulfonates. When the compounds contain acidic side chains, suitable pharmaceutically acceptable base addition salts of the compounds provided herein include, but are not limited to, metal salts made of aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or organic salts made of lysine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucosamine), and procaine.

[0170] In some embodiments, the compound of the present invention is an isotopically enriched apelelis, for example... 13 C1-Apeliximab 18 O1-Apeliximab 17 O1-Apeliximab 13 C1 18 O1-Apeliximab and / or 13 C1 17O1-Apelips. Apellis is a phosphatidylinositol-3-kinase (PI3K) inhibitor with primary PI3Kα inhibitory activity. In in vitro and in vivo models, gain-of-function mutations in the gene encoding the catalytic α-subunit of PI3K (PIK3CA) lead to activation of PI3Kα and Akt signaling, cell transformation, and tumorigenesis. In breast cancer cell lines, apellis inhibits phosphorylation of downstream targets of PI3K, including Akt, and exhibits activity in cell lines containing PIK3CA mutations. In vivo, apellis inhibits the PI3K / Akt signaling pathway and reduces tumor growth in xenograft models, including breast cancer models. Apellis treatment for PI3K inhibition has been shown to induce increased estrogen receptor (ER) transcription in breast cancer cells. In xenograft models derived from ER-positive PIK3CA-mutated breast cancer cell lines, the combination of apelips and fulvestrant showed enhanced antitumor activity compared to treatment alone. Apelips has been approved by the FDA as a Piqray drug. TM For example, it can be combined with fulvestrant to treat postmenopausal women and men with HR-positive, HER2-negative, PIK3CA-mutated, advanced or metastatic breast cancer.

[0171] In one specific embodiment, the compound of the present invention is 18 O1-Apeliximab (also referred to as Apeliximab in this article) 18 O1). 18 O1-apelis is the stable isotopic form of apelis, in which the oxygen atom at the proline amide position is enriched with oxygen-18 or... 18 O. The chemical name of the compound is (2S)-N. 1 -[4-methyl-5-[2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridinyl]-2-thiazolyl]-1,2-pyrrolidinedicarboxamide. The compound has the structure shown herein:

[0172]

[0173] In some implementations, isotope enrichment of apelix (e.g. 18The isotopic enrichment level of O1-apelis is about 90% or higher, about 91% or higher, about 92% or higher, about 93% or higher, about 94% or higher, about 95% or higher, about 96% or higher, about 97% or higher, about 98% or higher, about 99% or higher, or 100%. In one embodiment, an isotopic enrichment level of 95% or higher is provided. 18 O-isotope enrichment 18 O1-Apeliximab. In one implementation, it is provided with a content of 94% or higher. 18 O-isotope enrichment 18 O1-apelis. In one embodiment, the isotope enrichment level is determined using mass spectrometry.

[0174] In some implementation schemes, apeliximab- 18 O1 uses H2 18 O or 18 O-water as 18 It is prepared from the source of O. Apellis- 18 O1 can be used for, but is not limited to, examples, but not limited to Figure 5 The synthetic route shown in the figure is used to prepare it. This synthetic route ensures that only isotope enrichment is achieved. 18 O-water will be incorporated into the formamide-substituted pyrrolidine ring of apeliximab. In this case, the isotopic enrichment level in the compound may be equal to or slightly lower than that of apeliximab. 18 The enrichment level of O-water isotopes. For example, 18 O-water usually contains a certain level of 16 O and 17 O, for example, about 1% to 1.5% 16 O and similar or even higher levels 17 O, using this method, the isotope will be carried to 18 O-apelis. Therefore, the obtained apelisi- 18 O1 will have, for example, ≥95% (±0.5%) 18 O-isotope enrichment, ≥96% (±0.5%) 18 O-isotope enrichment, ≥97% (±0.5%) 18 O-isotope enrichment, ≥98% (±0.5%) 18 O- isotope enrichment or ≥99% (±0.5%) 18 O- isotope enrichment. In one specific implementation, the obtained apelistat- 18 O1 has ≥95% (±0.5%) 18 O- isotope enrichment. In one implementation, apeliximab- 18 The molecular weight of O1 is 443.45. (Apeliximab -) 18The preparation of O1 is further described in Example 1 below.

[0175] Composition

[0176] In one embodiment, a pharmaceutical composition is provided comprising an isotopically enriched compound as described herein (e.g., compounds of formulas I to III or those in Table 1) or a pharmaceutically acceptable salt, ester, chelate, hydrate, or solvate thereof, and a pharmaceutically acceptable carrier. In another embodiment, a pharmaceutical composition is provided comprising a compound of formulas I to III or those in Table 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In yet another embodiment, a pharmaceutical composition is provided comprising a compound of formulas I to III or those in Table 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, provided that the compound is not niraparib. In yet another embodiment, a pharmaceutical composition is provided comprising isotopically enriched apeliximab (e.g., niraparib). 18 O1-apelis and pharmaceutically acceptable carriers.

[0177] Pharmaceutical compositions can be prepared as is known in the art (see, for example, Remington: The Science and Practice of Pharmacy, 20th edition, 2000). For example, therapeutic compounds and / or compositions, together with one or more solid or liquid drug carrier substances and / or additives (or excipients), and, if desired, with other pharmaceutically active compounds having therapeutic or preventative effects, can be formulated into suitable forms or dosage forms for use as medicines in humans or veterinary medicines. Pharmaceutical formulations may also contain additives, many of which are known in the art, such as fillers, disintegrants, binders, lubricants, wetting agents, stabilizers, emulsifiers, dispersants, preservatives, sweeteners, colorants, flavoring agents, aromatizers, thickeners, diluents, buffers, solvents, solubilizers, agents for achieving depot function, salts for altering osmotic pressure, coating agents, or antioxidants.

[0178] Any suitable pharmaceutically acceptable carrier, diluent, excipient, excipient, or loading agent may be used in the compositions provided herein, such as preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aroma agents, antibacterial agents, antifungal agents, lubricants, and dispersants, depending on the method of application and the nature of the dosage form.

[0179] Some examples of suspending agents include ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum hydroxide, bentonite, agar, and tragacanth, or mixtures thereof. Prevention of microbial action can be ensured by a variety of antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Isotonic agents, such as sugars and sodium chloride, may also be desired. Prolonged absorption of injectable drug forms can be achieved by using agents with delayed absorption (e.g., aluminum monostearate and gelatin). Some examples of suitable carriers, diluents, solvents, or loading agents include water, ethanol, polyols, suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters such as ethyl oleate. Some examples of excipients include lactose, milk sugar, sodium citrate, calcium carbonate, and dicalcium phosphate. Some examples of disintegrants include starch, alginate, and certain complex silicates. Some examples of lubricants include magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol.

[0180] Pharmaceutically acceptable carriers may include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonics, and absorption delay agents. In one embodiment, the carrier is suitable for oral administration. Alternatively, the carrier may be suitable for intravenous, intraperitoneal, intramuscular, sublingual, or parenteral administration. In other embodiments, the carrier is suitable for surface application or inhalation administration. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the provisional preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is well known in the art. The use of any conventional media or agent in the pharmaceutical compositions provided herein is also contemplated unless it is incompatible with the active compound.

[0181] The pharmaceutical compositions described herein can be administered orally, for example, in the form of pills, tablets, lacquered tablets, sugar-coated tablets, granules, hard gelatin capsules and soft gelatin capsules, aqueous solutions, alcoholic or oily solutions, syrups, emulsions or suspensions, or rectally, for example, as suppositories. Administration can also be parenteral, for example, subcutaneously, intramuscularly or intravenously, in the form of solutions for injection or infusion. Other suitable forms of administration are, for example, percutaneous or topical application, such as in the form of ointments, creams, tinctures, sprays or percutaneous therapeutic systems; or inhalation, in the form of nasal sprays or aerosol mixtures; or, for example, microcapsules, implants or wafers.

[0182] In some embodiments, the pharmaceutical compositions provided herein are suitable for oral administration. For example, the pharmaceutical compositions may be in the following forms: hard-shell gelatin capsules, soft-shell gelatin capsules, flat capsules, pills, tablets, lozenges, powders, granules, soft lozenges, or sugar-coated pills. In one specific embodiment, the pharmaceutical composition is in the form of a tablet.

[0183] Alternatively, the pharmaceutical composition may be in the form of a solution, an aqueous liquid suspension, a non-aqueous liquid suspension, an oil-in-water liquid emulsion, an oil-in-water liquid emulsion, an elixir, or a syrup. The pharmaceutical composition may or may not be enterically coated. In some embodiments, the pharmaceutical composition is formulated for controlled release, such as delayed or prolonged release.

[0184] In other embodiments, the compounds and their compositions may be formulated in multiple dosage forms, i.e., multiple particulate dosage forms (e.g., hard gelatin capsules or conventional tablets prepared using a rotary tableting machine), which contain one or more beads or minitab groups for oral administration. Conventional tablets disperse rapidly upon entering the stomach. One or more coated beads or minitab groups may be compressed into tablets together with suitable excipients (e.g., binders, diluents / fillers, and disintegrants for conventional tablets).

[0185] Tablets, pills, beads, or minitabs of compounds and compositions thereof may be coated or otherwise compounded to provide dosage forms with controlled release, including delayed or prolonged release advantages, or to protect against acidic conditions of the stomach. For example, tablets or pills may comprise an internal dose component and an external dose component, the latter being in the form of a coating over the former. These two components may be separated by a polymer layer that controls the release of the internal dose.

[0186] In some embodiments, the layer may comprise at least one enteric polymer. In other embodiments, the layer may comprise at least one enteric polymer in combination with at least one water-insoluble polymer. In still other embodiments, the layer may comprise at least one enteric polymer in combination with at least one water-soluble polymer. In yet another embodiment, the layer may comprise at least one enteric polymer in combination with a pore-forming agent.

[0187] In some embodiments, the layer may comprise at least one water-insoluble polymer. In still other embodiments, the layer may comprise at least one water-insoluble polymer in combination with at least one water-soluble polymer. In yet another embodiment, the layer may comprise at least one water-insoluble polymer in combination with a pore-forming agent.

[0188] Some representative examples of water-soluble polymers include polyvinylpyrrolidone (PVP), hydroxypropyl methylcellulose (HPMC), hydroxypropylcellulose (HPC), and polyethylene glycol.

[0189] Representative examples of enteric polymers include cellulose esters and their derivatives (cellulose acetate phthalates, hydroxypropyl methylcellulose phthalates, hydroxypropyl methylcellulose acetate succinate), polyvinyl acetate phthalates, pH-sensitive methacrylate-methyl methacrylate copolymers, and shellac. These polymers can be used as dry powders or aqueous dispersions. Some commercially available materials are methacrylate copolymers manufactured by Rohm Pharma and sold under the trademarks Eudragit (LI00, SI 00, L30D), Cellacefate (cellulose acetate phthalates) from Eastman Chemical Co., Aquateric (aqueous dispersion of cellulose acetate phthalates) from FMC Corp., and Aqoat (aqueous dispersion of hydroxypropyl methylcellulose acetate succinate) from Shin Etsu KK.

[0190] Some representative examples of available water-insoluble polymers include ethyl cellulose, polyvinyl acetate (e.g., Kollicoat SR#30D from BASF), cellulose acetate, cellulose acetate butyrate, neutral copolymers based on ethyl acrylate and methyl methacrylate, and copolymers of acrylates and methacrylates with quaternary ammonium groups such as Eudragit NE, RS and RS30D, RL or RL30D, etc.

[0191] Any of the above-described polymers may be further plasticized with one or more pharmaceutically acceptable plasticizers. Some representative examples of plasticizers include triacetin, tributyl citrate, triethyl citrate, diethyl acetyl tri-n-butyl citrate phthalate, castor oil, dibutyl sebate, acetylated monoglycerides, and mixtures thereof. When used, the plasticizer may be based on about 3 to 30% by weight of the polymer, and more typically about 10 to 25% by weight. The type and content of the plasticizer depend on one or more polymers and the nature of the coating system (e.g., water- or solvent-based, solution- or dispersion-based, and total solids).

[0192] Pharmaceutical compositions must generally be sterile and stable under preparation and storage conditions. Compositions can be formulated as solutions, microemulsions, liposomes, or other ordered structures suitable for high drug concentrations. The carrier can be a solvent or dispersion medium comprising, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Appropriate flowability can be maintained, for example, by using coatings (e.g., lecithin), by maintaining the desired particle size in the case of dispersions, and by using surfactants. In many cases, isotonic agents, such as sugars, polyols (e.g., mannitol, sorbitol), or sodium chloride, will be preferably included in the composition. Prolonged absorption of injectable compositions can be achieved by including agents with delayed absorption (e.g., monostearate and gelatin) in the composition. Furthermore, compounds can be administered in time-release formulations, such as compositions containing slow-release polymers. Compounds can be prepared using carriers that will prevent rapid release, such as controlled-release formulations, including implants and microencapsulated delivery systems. Biodegradable and biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoester, polylactic acid, and polylactic acid-polyglycolic acid copolymer (PLG) can be used.

[0193] Many methods for preparing such formulations are generally known to those skilled in the art. Sterile injectable solutions can be prepared by incorporating an active compound (e.g., compounds of formulas I to III or those in Table 1) in the desired amount with one or a combination of the ingredients listed above (as needed) into a suitable solvent, followed by filtration and sterilization. Typically, dispersions are prepared by incorporating the active compound into a sterile carrier comprising a basic dispersion medium and other desired components from those listed above. In the case of sterile powders used to prepare sterile injectable solutions, typical preparation methods include vacuum drying and freeze-drying, which produce a powder containing the active ingredient plus any additional desired components from its previously sterile filtered solution. Compounds can also be formulated with one or more additional compounds to enhance their solubility.

[0194] For ease of administration and dosage uniformity, it is generally advantageous to formulate compositions in unit dosage form (e.g., parenteral compositions). The term "unit dosage form" refers to a physically discrete unit suitable as a single dose for use in human subjects and other animals, each unit containing a predetermined amount of active substance associated with a suitable drug carrier, calculated to produce the desired therapeutic effect. The specifications of the unit dosage form of the present invention may vary and are indicated by and directly dependent on: (a) the unique characteristics of the therapeutic compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations in the field of using such therapeutic compounds for the prevention or treatment of a target disease, such as PI3K-mediated diseases, proliferative diseases such as melanoma, colorectal adenoma, breast cancer, or pancreatic cancer. Dosage is discussed further below.

[0195] In some embodiments, a pharmaceutical composition is provided comprising a therapeutically effective amount of the compound and / or composition described herein, and a pharmaceutically acceptable carrier. In one embodiment, a pharmaceutical composition is provided for treating or preventing PI3K-mediated diseases, comprising the compound described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In another embodiment, a pharmaceutical composition is provided for preventing or treating PI3K-mediated diseases, such as proliferative diseases like melanoma, colorectal adenoma, breast cancer, or pancreatic cancer, wherein the composition comprises the compound described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In some such embodiments, the compound is... 18 O1-Apeliximab

[0196] Additional active compounds may also be incorporated into the compositions provided herein. For example, the pharmaceutical compositions provided herein may also contain at least one additional therapeutic agent, as discussed below. In one embodiment, a pharmaceutical composition is provided comprising at least one compound of the present invention, and a pharmaceutically acceptable carrier suitable for administration alone or in combination with one or more additional therapeutic agents to a human or animal subject. In some embodiments, the additional therapeutic agent is an anticancer agent, such as fulvestrant. In one specific embodiment, the compound of the present invention is... 18 O1-apelis and at least one other therapeutic agent is fulvestrant.

[0197] Therefore, combined pharmaceutical compositions are provided, for example, for use in any of the methods described herein, wherein the pharmaceutical composition comprises the compound of the present invention associated with a pharmaceutically acceptable carrier in free or pharmaceutically acceptable salt form. The combined pharmaceutical composition may comprise the compound of the present invention in free or pharmaceutically acceptable salt form as an active ingredient, one or more pharmaceutically acceptable carrier substances, and optionally one or more other pharmaceutical substances. Such combined pharmaceutical compositions may be in the form of a single dose unit or as a kit. Combined pharmaceutical compositions comprising a therapeutically effective amount of the compound of the present invention in free or pharmaceutically acceptable salt form and a second pharmaceutical substance are intended for simultaneous or sequential administration.

[0198] "Combination" refers to a fixed combination in the form of a single dose unit, or a kit for combined administration, wherein the compounds of the present invention and the combination partner (i.e., additional therapeutic agents or drugs as further described below) may be administered independently at the same time or separately at time intervals, particularly wherein these time intervals allow the combination partner to exhibit synergistic effects, such as a co-administration. The terms "co-administration" or "combination administration" as used herein are intended to cover the administration of selected combination partners to a single subject (e.g., a patient) in need, and are intended to include treatment regimens in which the agents are not necessarily administered via the same route of administration or simultaneously. The term "drug combination" as used herein refers to a product resulting from mixing or combining more than one active ingredient, and includes both fixed and non-fixed combinations of active ingredients. The term "fixed combination" means that the active ingredients (e.g., the compounds of the present invention and the combination partner) are both administered simultaneously to the patient in the form of a single entity or dose. The term "non-fixed combination" means that the active ingredients (e.g., the compounds of the present invention and the combination partner) are both administered simultaneously, concurrently, or sequentially to the patient as separate entities without a specific time limit, wherein such administration provides therapeutically effective levels of both compounds in the patient. The latter also applies to cocktail therapy, such as the application of three or more active ingredients.

[0199] The amount of the compounds of the present invention in a formulation can vary within the full range used by those skilled in the art. Typically, the formulation will contain about 0.01 to 99.99% by weight of the compounds of the present invention based on a weight percentage (wt%) of the total formulation, wherein the balance is one or more suitable pharmaceutical excipients. In some embodiments, the compounds are present at a level of about 1 to 80% by weight. Dosage is discussed further below.

[0200] The present invention also provides a pillbox containing one or more compounds of the present invention and optionally combinations thereof disclosed herein. In one embodiment, the pillbox contains a compound or composition of the present invention along with a packaging insert or other label including instructions for use thereof. The pillbox may optionally contain one or more additional components, such as acids, bases, buffers, inorganic salts, solvents, antioxidants, preservatives, or metal chelators; a device for delivery and administration, such as a syringe, vial, etc.; and instructions for use thereof.

[0201] How to use

[0202] This invention provides a method for treating or preventing disease, comprising administering the compounds and compositions of this invention to a subject in need. It is not intended to be theoretically limited, but it is believed that the isotope-enriched compounds provided herein can improve their therapeutic efficacy by improving their therapeutic biodistribution and / or pharmacokinetic profile, for example by increasing the bioavailability of the compound, reducing the metabolism of the compound, improving the stability of the compound, and / or altering the release rate of the active compound from a prodrug.

[0203] In one embodiment, the present invention relates to a method for modulating the metabolic or pharmacokinetic profile of an amide-containing (i.e., amide-functionalized) compound in a subject, comprising administering to the subject an isotopically enriched compound and / or pharmaceutical composition as described herein, wherein the metabolic or pharmacokinetic profile of the amide-containing compound is modulated compared to administration of the same compound having only naturally abundant isotopes (i.e., non-isotopically enriched compounds). In some embodiments, methods are provided for reducing the metabolism of a compound, reducing the therapeutic toxicity of a compound, reducing the adverse effects of a compound, improving the tolerability of a compound, improving the biodistribution of a compound, and / or improving the therapeutic or prophylactic effects of a compound in a subject, said methods comprising administering to the subject an isotopically enriched compound or pharmaceutical composition as described herein, wherein the isotopically enriched compound exhibits reduced metabolism, reduced therapeutic toxicity, reduced adverse effects, improved tolerability, improved biodistribution, and / or improved therapeutic or prophylactic effects compared to administration of a compound having only naturally abundant isotopes (i.e., non-isotopically enriched compounds).

[0204] In one exemplary embodiment, the present invention relates to a method of modulating (e.g., reducing) apelips metabolism, comprising administering to a subject an isotopically enriched apelips compound and / or a pharmaceutical composition thereof as described herein, wherein the metabolism of the isotopically enriched apelips compound is modulated compared to administration of the same compound having only naturally abundant isotopes (i.e., a non-isotopically enriched compound). In some such embodiments, isotopically enriched apelips is 18 O1-Apeliximab. In one implementation scheme... 18 O1-Apeliximab is approximately 93%.18 O1-enriched, approximately 94% 18 O1-enriched, approximately 95% 18 O1-enriched, approximately 96% 18 O1-enriched, approximately 97% 18 O1-enriched, approximately 98% 18 O1-enriched or approximately 99% 18 O1 enriched. In one implementation, 18 O1-apelis is administered in combination with at least one other therapeutic agent (e.g., an anticancer drug, such as fulvestrant).

[0205] In one embodiment, the present invention relates to treating PI3K-mediated diseases, i.e., diseases or conditions improved by inhibiting phosphoinositol 3-kinase (PI3K). In some embodiments, phosphoinositol 3-kinase (PI3K)-mediated diseases are PI3Kα-mediated diseases (or diseases mediated by overexpression or amplification of PI3Kα, somatic mutations of PI3K, germline or somatic mutations of PTEN, or mutations and translocations of p85α for upregulating the p85-p110 complex), particularly conditions that respond beneficially to inhibition of PI3K kinase, particularly inhibition of PI3Kα or its mutant forms.

[0206] In some implementations, PI3K-mediated diseases are proliferative disorders, such as tumor or cancer cell growth. PI3K-mediated diseases may include those exhibiting PI3Kα overexpression or amplification, somatic mutations in PIK3CA, germline or somatic mutations in PTEN, or mutations and translocations of p85α that upregulate the p85-p110 complex. In some specific embodiments, the compounds of the present invention can be used to treat cancers in humans or animals (e.g., mice), including, for example: sarcoma; lung; bronchus; prostate; breast (including sporadic breast cancer and Cowden's disease); pancreas; gastrointestinal cancer; colon; rectum; colon cancer; colorectal adenoma; thyroid gland; liver; intrahepatic bile ducts; hepatocytes; adrenal gland; stomach; gastric region; glioma; glioblastoma; endometrium; melanoma; kidney; renal pelvis; bladder; uterine body; cervix; vagina; ovary; multiple myeloma; esophagus; leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; brain; brain cancer; oral cavity and pharynx; larynx; small intestine; non-Hodgkin lymphoma; melanoma; villous colonic adenoma; tumor formation; epithelial-characteristic tumor formation; lymphoma; breast cancer (mammary). (carcinoma); basal cell carcinoma; squamous cell carcinoma; actinic keratosis; neoplastic diseases, including solid tumors; tumors of the neck or head; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; and Walden stroem's disease.

[0207] In other implementations, PI3K-mediated diseases are selected from: polycythemia vera, essential thrombocythemia, myelofibrosis with myeloid metaplasia, asthma, COPD, ARDS, Loffler's syndrome, eosinophilic pneumonia, parasitic (especially metazoan) infections (including tropical eosinophilia), bronchopulmonary aspergillosis, polyarteritis nodosa (including Churg-Strauss syndrome), eosinophilic granuloma, eosinophilic-associated diseases affecting the airways due to drug reactions, psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, scleroderma, vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphigus, acquired epidermolysis bullosa, autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure erythrocytic anemia, and idiopathic thrombocytopenic purpura), and systemic erythema. Lupus, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Steven-Johnson syndrome, idiopathic stomatitis, autoimmune inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease), endocrine ophthalmopathy, Graves' disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonia, multiple sclerosis, primary biliary cirrhosis, uveitis (pre- and post-), ​​interstitial pulmonary fibrosis, psoriatic arthritis, glomerulonephritis, cardiovascular disease, atherosclerosis, hypertension, deep vein thrombosis, stroke, myocardial infarction, unstable angina, thromboembolism, pulmonary embolism, thrombolysis, acute arterial ischemia, peripheral thromboocclusion, and coronary artery disease, reperfusion injury, retinopathy (e.g., diabetic retinopathy or hyperbaric oxygen-induced retinopathy), and conditions characterized by aqueous humor secretion or increased intraocular pressure (e.g., glaucoma).

[0208] In some implementations, PI3K-mediated diseases are proliferative diseases, such as melanoma, colorectal adenoma, breast cancer, and / or pancreatic cancer. In some such implementations, the breast cancer is hormone receptor-positive (HR+) and human epidermal growth factor receptor-negative (HER2-). In some implementations, the breast cancer is PIK3CA mutated.

[0209] For the purposes described above, the required dosage will naturally vary depending on the method of administration, the specific condition to be treated, and the desired effect. Generally, a systemic satisfactory result is obtained at a daily dose of about 0.03 to about 100.0 mg / kg of body weight, for example, about 0.03 to about 10.0 mg / kg of body weight. The specified daily dose for humans is typically conveniently administered in the range of about 0.5 mg to about 3 g (e.g., about 5 mg to about 1.5 g), for example, in divided doses up to four times a day, or in a delayed form, or once a day. Suitable unit dosage forms for oral administration include about 0.1 to about 500 mg, for example, about 1.0 to about 500 mg of active ingredient, such as 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 500 mg of active ingredient. In one embodiment, the compound of the invention is administered at a daily dose of about 300 mg. In some such embodiments, the compound is administered once daily. In some such implementations, the compound is administered orally once daily with food.

[0210] The compounds of the present invention can be administered via any conventional route, particularly via enteral (e.g., orally, in tablet or capsule form), parenteral (e.g., in injectable solutions or suspensions), or topically (e.g., in the form of lotions, gels, ointments, or creams, via inhalation, intranasal administration, or suppositories). In one specific embodiment, the compounds are administered orally in tablet form.

[0211] The compounds of the present invention can be administered in free form or as pharmaceutically acceptable salts. As discussed above, such salts can be prepared in a conventional manner and generally exhibit the same activity sequence as the free compounds.

[0212] The PI3K pathway is a central oncogenic pathway regulating cell proliferation, metabolism, growth, survival, and apoptosis. Activation of downstream PI3K signaling is crucial for mediating the transformative potential of oncogenes and tumor suppressors in many tumor types. Dysregulation of PI3K signaling is associated with resistance to various therapies, including anti-estrogens, trastuzumab, radiotherapy, and chemotherapy. PIK3CA mutations have been reported in up to 43.3% of HR-positive, HER2-negative tumors (Wan et al., 2018). These mutations increased PI3K activity and induced breast tumor formation in transgenic mice (Zhao and Vogt, 2008; Meyer et al., 2011; Miller et al., 2011).

[0213] Therefore, methods for preventing or treating symptoms, disorders, or diseases mediated by PI3K enzyme activation are also provided, for example in subjects requiring such treatment, wherein the method comprises administering an effective amount of the compound of the invention (e.g., 18 O1-apelis) or a pharmaceutically acceptable salt thereof, wherein the compound of the present invention or a pharmaceutically acceptable salt thereof is, for example, in a free form or a pharmaceutically acceptable salt form as a drug in any of the methods shown herein. 18 O1-apelis inhibits PI3K primarily by inhibiting PI3Kα; gain-of-function mutations in PIK3CA lead to activation of PI3Kα and protein kinase B (Akt) signaling, cell transformation, and tumorigenesis in in vitro and in vivo models.

[0214] Evidence has also emerged that combinations of PI3K inhibitors with inhibitors of other pathways could be used to treat cancer and proliferative diseases in humans. Therefore, methods for preventing or treating conditions, disorders, or diseases mediated by PI3K enzyme activation are also provided, for example in subjects requiring such treatment, wherein the method comprises administering an effective amount of the compound of the invention (e.g., ...) to said subject. 18 O1-apelis) or a pharmaceutically acceptable salt thereof, wherein the compound of the present invention or a pharmaceutically acceptable salt thereof is, for example, in a free form or a pharmaceutically acceptable salt form as a drug in any of the methods described herein, and the method further includes administration of at least one additional anticancer agent as described herein.

[0215] Although breast cancer affects a single anatomical site, it is a heterogeneous disease with different phenotypic manifestations (Perou et al., 2000). The identification of different biological subtypes is mainly carried out using immunohistochemistry (Nielsen et al., 2004) and gene expression profiling (Perou et al., 2000), in which breast cancer is divided into three biological subgroups: those expressing estrogen receptor (ER), those expressing HER2 (with or without ER expression), and those expressing neither of these nor progesterone receptor (PR; triple negative) (Fragomeni et al., 2018). Approximately 70% of invasive breast cancers in women older than 45 years express ER and / or PR but not HER2, and are termed HR-positive, HER2-negative (Huang et al., 2005). Although the most common breast cancer subtype in men is HR-positive, HER2-negative (Anderson et al., 2004; Ottini et al., 2010), little is known about the biology of male breast cancer due to the low prevalence of the disease in men and the historical exclusion of men from breast cancer clinical trials.

[0216] Patients with ER-positive metastatic breast cancer often respond to endocrine therapy alone or in combination with targeted therapy, which reduces tumor burden and symptoms with fewer side effects and toxicities compared to chemotherapy (Ma and Sparano, 2020). Adrenal progenitors, testosterone, and androstenedione are converted to estradiol and estrone via aromatase activity, among which aromatase-specific inhibitors (anastrozole, letrozole, and exemestane) have shown efficacy in depleting estrogen; additionally, tamoxifen, which works by interfering with ER signaling, is an effective treatment option (Ma and Sparano, 2020).

[0217] In cases where the primary goal of treatment is palliative care, modern endocrine therapy prolongs disease progression but does not provide a cure (Ma and Sparano, 2020). However, endocrine resistance is a major concern, with distinctions between primary (de novo) and secondary (occurring during treatment) resistance; only 50% of patients with ER-positive breast cancer benefit from first-line endocrine therapy, and all initially responsive patients acquire resistance, leading to tumor recurrence and growth (Fu et al., 2013). Even in patients with secondary endocrine resistance, progression-free survival is short in the case of endocrine-based treatment options. A crucial consideration when deciding whether to continue with another line of endocrine therapy or switch to chemotherapy when a patient's cancer is unresponsive or has ceased to respond to a given line of endocrine therapy or targeted therapy is the PIK3CA mutation status (Ma and Sparano, 2020).

[0218] Approximately 20% to 30% of breast cancer patients overexpress the Her-2 / neu-ErbB2 target of the drug trastuzumab. Although trastuzumab has shown durable responses in some patients expressing Her2 / neu-ErbB2, only a subset of these patients respond. Recent work has shown that this limited response rate can be significantly improved by combining trastuzumab with inhibitors of the PI3K or PI13K / AKT pathways (Chan et al., Breast Can. Res. Treat. 91:187 (2005), Woods Ignatoski et al., Brit. J. Cancer 82:666 (2000), Nagata et al., Cancer Cell 6:117 (2004)).

[0219] Many human malignancies express activating mutations or elevated levels of Her1 / EGFR, and numerous antibodies and small molecule inhibitors targeting this receptor tyrosine kinase have been developed, including tarceva, gefitinib, and erbitux. However, while EGFR inhibitors have shown antitumor activity in some human tumors (e.g., NSCLC), they have failed to improve overall patient survival in all patients with EGFR-expressing tumors. This can be justified by the fact that many downstream targets of Her1 / EGFR are mutated or dysregulated at high frequencies in a variety of malignancies involving the PI3K / Akt pathway. For example, gefitinib inhibits the growth of adenocarcinoma cell lines in in vitro assays. Nevertheless, subclones of these cell lines that are resistant to gefitinib can be selected, indicating elevated activation of the PI3 / Akt pathway. Downregulation or inhibition of this pathway sensitizes the resistant subclones to gefitinib (Kokubo et al., Brit. J. Cancer 92:1711 (2005)). Furthermore, in an in vitro model of breast cancer with PTEN mutations and EGFR overexpression, inhibition of both the PI3K / Akt pathway and EGFR produced a synergistic effect (She et al., Cancer Cell 8:287-297 (2005)). These results suggest that the combination of gefitinib and PI3K / Akt pathway inhibitors would be an attractive therapeutic strategy for cancer.

[0220] In glioblastoma xenograft models, the combination of AEE778 (an inhibitor of Her-2 / neu / ErbB2, VEGFR and EGFR) and RAD001 (an inhibitor of mTOR, a downstream target of Akt) produced greater combined potency than any single drug (Goudar et al., Mol. Cancer. Ther. 4:101-112 (2005)).

[0221] Anti-estrogens (e.g., tamoxifen) inhibit breast cancer growth by inducing cell cycle arrest, which requires the action of the cell cycle inhibitor p27Kip. Activation of the Ras-Raf-MAP kinase pathway has been shown to alter the phosphorylation state of p27Kip, thereby reducing its inhibitory activity in cell cycle arrest and contributing to anti-estrogenic resistance (Donovan et al., J. Biol. Chem. 276:40888, (2001)). As reported by Donovan et al., inhibition of MAPK signaling by treatment with a MEK inhibitor reversed the aberrant phosphorylation state of p27 in hormone-refractory breast cancer cell lines, restoring hormone sensitivity in doing so. Similarly, phosphorylation of p27Kip by Aid also eliminated its cell cycle arrest effect (Viglietto et al., Nat. Med. 8:1145 (2002)).

[0222] Therefore, in another embodiment, the compound of the present invention (e.g., 18 O1-apelis is used to treat hormone-dependent cancers, such as breast and prostate cancer. In this application, it aims to reverse the hormone resistance common in these cancers using conventional anticancer agents.

[0223] In hematologic malignancies such as chronic myelogenous leukemia (CML), chromosomal translocations are the cause of constitutive activation of the BCR-Abl tyrosine kinase. As a result of Abl kinase activity inhibition, patients respond to the small-molecule tyrosine kinase inhibitor imatinib. However, many patients with advanced disease who initially respond to imatinib subsequently relapse due to resistance-conferred mutations in the Abl kinase domain. In vitro studies have shown that BCR-Abl utilizes the Ras-Raf kinase pathway to elicit its action. Furthermore, inhibition of more than one kinase in the same pathway provides additional protection against resistance-conferred mutations.

[0224] Therefore, in another embodiment, the compound of the present invention (e.g., 18 O1-apelixir and at least one kinase inhibitor selected from Gleevec TM Other combinations of drugs are used to treat hematologic cancers, such as chronic myeloid leukemia (CML). Through this use, it is intended to reverse or prevent resistance to at least one of the other drugs.

[0225] Since activation of the PI3K / Akt pathway drives cell survival, its inhibition, combined with apoptosis-driving therapies (including radiotherapy and chemotherapy) in cancer cells, elicits an improved response (Ghobrial et al., CA Cancer J. Clin 55:178-194 (2005)). As an example, the combination of PI3 kinase inhibitors with carboplatin has shown synergistic effects in both in vitro proliferation and apoptosis assays, as well as in in vivo tumor potency in an ovarian cancer xenograft model (Westfall and Skinner, Mol. Cancer Ther. 4:1764-1771 (2005)).

[0226] Beyond cancer and proliferative disorders, the existence of inhibitors of class 1A and class 1B PI3 kinases provides accumulating evidence of therapeutic availability in other disease areas. Inhibition of the PI3K isotype product p110β of the PIK3CB gene has been shown to involve cleavage-induced platelet activation (Jackson et al., Nature Medicine 11:507-514 (2005)). Therefore, PI3K inhibitors that inhibit p110β could be used as single agents or in combination in antithrombotic therapy. The PIK3CD gene product, isoform p110δ, is important in B cell function and differentiation (Clayton et al., J. Exp. Med. 196:753-763 (2002)), T cell-dependent and T cell-independent antigen responses (Jou et al., Mol. Cell. Biol. 22:8580-8590 (2002)), and mast cell differentiation (Ali et al., Nature 431:1007-1011 (2004)). Therefore, p110δ inhibitors are expected to be used to treat B cell-driven autoimmune diseases and asthma. Finally, inhibition of the PI3KCG gene isotype product p110γ resulted in a reduced T-cell response rather than a B-cell response (Reif et al., J. Immunol. 173:2236-2240 (2004)), and its inhibition was demonstrated to be effective in animal models of autoimmune diseases (Camps et al., Nature Medicine 11:936-943 (2005), Barber et al., Nature Medicine 11:933-935 (2005)).

[0227] In another embodiment, a method is provided for treating a human or animal subject suffering from a proliferative disease (e.g., cancer). A method is provided for treating a human or animal subject requiring such treatment, comprising administering to the subject a therapeutically effective amount of the compound of the invention (e.g., alone or in combination with one or more other anticancer agents).18 O1-Apeliximab). In particular, the composition will be formulated together as a combination therapy or administered alone. Suitable anticancer agents for use with the compounds of the present invention include, but are not limited to, one or more compounds selected from: kinase inhibitors, antiestrogens, antiandrogens, other inhibitors, cancer chemotherapy drugs, alkylating agents, chelating agents, biological response modulators, cancer vaccines, and agents for antisense therapy, as further described below.

[0228] Kinase inhibitors used in combination with the compounds of the present invention as anticancer agents include, but are not limited to: epidermal growth factor receptor (EGFR) kinase inhibitors, such as small molecule quinazolines, such as gefitinib (US Patent Nos. 5,457,105, 5,616,582 and 5,770,599), ZD-6474 (WO 01 / 32651), erlotinib (Tarceva™, US Patent Nos. 5,747,498 and WO 96 / 30347), and lapatinib (US Patent Nos. 6,727,256 and WO 02 / 02552); vascular endothelial growth factor receptor (VEGFR) kinase inhibitors, including SU-11248 (WO 01 / 60814), SU 5416 (US Patent Nos. 5,883,113 and WO 02 / 60814). 99 / 61422), SU 6668 (US Patent No. 5,883,113 and WO 99 / 61422), CHIR-258 (US Patent No. 6,605,617 and US Patent No. 6,774,237), vatalanib or PTK-787 (US Patent No. 6,258,812), VEGF-Trap (WO 02 / 57423), B43-genistein (WO-09606116), retinoic acid (p-hydroxyaniline retinoic acid) (US Patent No. 4,323,581), IM-862 (WO 02 / 62826), bevacizumab or Avastin TM (WO 94 / 10202), KRN-951, 3-[5-(methylsulfonylpiperidinylmethyl)-indolyl]-quinolone, AG-13736 and AG-13925, pyrrolo[2,1-t][1,2,4]triazine, ZK-304709, Veglin TMVMDA-3601, EG-004, CEP-701 (US Patent No. 5,621,100), Candy (WO 04 / 09769); Erb2 tyrosine kinase inhibitors, such as pertuzumab (WO 01 / 00245), trastuzumab, and rituximab; Aid protein kinase inhibitors, such as RX-0201; Protein kinase C (PKC) inhibitors, such as LY-317615 (WO 95 / 17182) and perifosine (US 2003171303); Raf / Map / MEK / Ras kinase inhibitors, including sorafenib (BAY 43-9006), ARQ-350RP, LErafAON, BMS-354825AMG-548, and others in WO The following are disclosed in 03 / 82272: Fibroblast growth factor receptor (FGFR) kinase inhibitors; cell-dependent kinase (CDK) inhibitors, including CYC-202 or roscovitine (WO 97 / 20842 and WO 99 / 02162); platelet-derived growth factor receptor (PDGFR) kinase inhibitors, such as CHIR-258, 3G3 mAb, AG-13736, SU-11248, and SU6668; and Bcr-Abl kinase inhibitors and fusion proteins, such as STI-571 or Gleevec. TM (Imatinib)

[0229] Estrogen-targeting agents (anti-estrogens) used in combination with the compounds of the present invention for anticancer therapy include, but are not limited to: selective estrogen receptor modulators (SERMs), including tamoxifen, toremifene, and raloxifene; aromatase inhibitors, including anastrozole. TM ); and estrogen receptor downregulators (ERDs), including fulvestrant (Faslodex). TM ).

[0230] Fulvestrant is a small molecule drug used to treat hormone receptor (HR)-positive metastatic breast cancer in postmenopausal women whose disease has progressed after anti-estrogen therapy. It is an estrogen receptor antagonist without agonist activity, acting by downregulating and degrading estrogen receptors. Currently, it is used in two ways: as monotherapy for breast cancer, and in combination with apelelis for the treatment of HR-positive, HER2-negative, PIK3CA-mutated advanced or metastatic breast cancer. In one specific embodiment, fulvestrant is used in combination with a compound of the present invention (e.g., 18O-apelelis) for the treatment of breast cancer, such as HR-positive, HER2-negative, PIK3CA-mutated advanced or metastatic breast cancer.

[0231] Androgen-targeting agents (anti-androgens) used in combination with the compounds of the present invention for anticancer treatment include, but are not limited to, flutamide, bicalutamide, finasteride, aminoglutethamide, ketoconazole, and corticosteroids.

[0232] Other inhibitors that can be used in combination with the compounds of the present invention as anticancer agents include, but are not limited to: protein farnesyltransferase inhibitors, including tipifarnib or R-115777 (US 2003134846 and WO 97 / 21701), BMS-214662, AZD-3409 and FTI-277; topoisomerase inhibitors, including merbarone and diflomotecan (BN-80915); kinesin spindle protein (KSP) inhibitors, including SB-743921 and MKI-833; and proteasome regulators, such as bortezomib or Velcade. TM (US Patent No. 5,780,454), XL-784; and cyclooxygenase 2 (COX-2) inhibitors, including nonsteroidal anti-inflammatory drugs (NSAIDs).

[0233] Cancer chemotherapy agents used in combination with the compounds of the present invention as anticancer agents include, but are not limited to, anastrozole (Arimidex). TM ), Bicalutamide (Casodex) TM ), Bleoxane sulfate TM MyleranTM Busulfex injection TM Capecitabine (Xeloda) TM N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin TM Carmustine (BiCNU) TM ), chlorambucil (Leukeran) TM ), cisplatin TM ), cladribine (Leustatin) TM ), Cytoxamide TM Or Neosar TM ), cytarabine, cytosine arabinoside (Cytosar-U) TM ), Cytarabine liposome injection (DepoCyt) TM ), dacarbazine (DTIC-Dome) TM Cosmegan (actinomycin D), daunorubicin hydrochloride (Cerubidine) TM), DaunoXome™ liposome citrate injection, Dexamethasone, Docetaxel (Taxotere™), Doxorubicin (Adriamycin™, Rubex™), Etoposide (Vepesid™), Fludarabine phosphate Phosphate (Fludara™), 5-Fluorouracil (Adrucil™, Efudex™), Flutamide (Eulexin™), Tezacitibine, Gemcitabine (Difluorodeoxycytidine), Hydroxyurea (Hydrea™), Idarubicin (Idamycin™), Ifosfamide (IFEXTM), Irinotecan (Camptosar™), L-Asparaginase (ELSPAR™), Calcium Leucovorin, Melphalan (Alkeran™), 6-Mercaptopurine (Purinethol™), Methotrexate (Folex™), Novantrone™, Mylotarg, Taxol™, Phoenin (Yttrium 90 / MX-DTPA), Pentostatin, Polyphenylpropanol 20 (Polifeprosan™) 20) Carmustine implant (Gliadel™), tamoxifen citrate (Nolvadex™), teniposide (Vumon™), 6-thioguanine, thiotepa, tirapazamine (Tirazone™), topotecan hydrochloride for injection (Hycamptin™), vinblastine (Velban™), vincristine (Oncovin) TM ) and Changchun Ruibin (NavelbineTM).

[0234] Alkylating agents used in conjunction with the compounds of the present invention include, but are not limited to: VNP-40101M or cloretizine, oxaliplatin (US Patent No. 4,169,846, WO 03 / 24978 and WO 03 / 04505), glufosfamide, mafosfamide, etopophos (US Patent No. 5,041,424), prednimustine; succinate; busulfan; irofluven (acylfulvene); penclomedine; pyrazoline acridine (PD-115934); O6-benzylguanine; decitabine. e)(5-aza-2-deoxycytidine); brostallicin; mitomycin C (MitoExtra); TLK-286 (Telcyta™); temozolomide; trabectedin (US Patent No. 5,478,932); AP-5280 (a platinum salt of cisplatin); porfiromycin; and clearazide (meclorethamine).

[0235] Chelating agents used in conjunction with the compounds of the present invention include, but are not limited to: tetrathiomolybdate (WO 01 / 60814); RP-697; chimeric T84.66 (cT84.66); and gadofosveset (Vasovist). TM ); deferoxamine; and bleomycin, optionally in combination with electroporation (EPT).

[0236] Biological response modulators (e.g., immunomodulators) used in combination with the compounds of the present invention include, but are not limited to: staurosprine and its macrocyclic analogs, including UCN-01, CEP-701, and midostaurin (see WO 02 / 30941, WO 97 / 07081, WO 89 / 07105, U.S. Patent No. 5,621,100, WO93 / 07153, WO 01 / 04125, WO 02 / 30941, WO 93 / 08809, WO 94 / 06799, WO 00 / 27422, WO 96 / 13506, and WO 88 / 07045); squalamine (WO 01 / 79255); DA-9601 (WO 98 / 04541 and US Patent No. 6,025,387); alemtuzumab; interferons (e.g., IFN-α, IFN-β, etc.); interleukins, especially IL-2 or aldesleukin, and IL-1, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 and their active biological variants having an amino acid sequence of greater than 70% of the natural human sequence; altretamine (Hexylen™); SU 101 or leflunomide (WO WO 04 / 06834 and U.S. Patent No. 6,331,555; imidazoquinolines, such as resiquimod and imiquimod (U.S. Patent Nos. 4,689,338, 5,389,640, 5,268,376, 4,929,624, 5,266,575, 5,352,784, 5,494,916, 5,482,936, 5,346,905, 5,395,937, 5,238,944 and 5,525,612); and SMIPs, including benzo[a]azole, anthraquinone, thiourea, and tryptanthrin (WO 04 / 87153, WO 04 / 64759 and WO 04 / 60308).

[0237] Anticancer vaccines used in combination with the compounds of the present invention include, but are not limited to: Avicine TM (Tetrahedron Lett. 26:2269-70(1974)); Ogovomica (OvaRex) TM Theratope TM(STn-KLH); melanoma vaccine; GI-4000 series (GI-4014, GI-4015, and GI-4016), which target five mutations in the Ras protein; GlioVax-1; MelaVax; Advexin TM Or IGN-201 (WO 95 / 12660); Sig / E7 / LAMP-1, encoding HPV-16E7; MAGE-3 vaccine or M3TK (WO 94 / 05304); HER-2VAX; ACTIVE, which stimulates tumor-specific T cells; GM-CSF cancer vaccine; and vaccines based on Listeria monocytogenes.

[0238] Anticancer agents for use in combination with the compounds of the present invention also include antisense compositions, such as, but not limited to: AEG-35156 (GEM-640); AP-12009 and AP-11014 (TGF-β2 specific antisense oligonucleotides); AVI-4126; AVI-4557; AVI-4472; oblimersen (Genasense™); JFS2; aprinocarsen (WO 97 / 29780); GTI-2040 (R2 ribonucleotide reductase mRNA antisense oligonucleotide) (WO 98 / 05769); GTI-2501 (WO98 / 05769); liposome-encapsulated c-Raf antisense oligodeoxynucleotide (LErafAON) (WO 98 / 43095); and Sima-027 (RNAi-based therapeutic targeting of VEGFR-1 mRNA).

[0239] The compounds of the present invention can also be combined with bronchodilator or antihistamine substances in pharmaceutical compositions. Such bronchodilators include, but are not limited to, anticholinergic or antimuscarinic agents, particularly ipratropium bromide, oxytropium bromide, and tiotropium bromide, and β-2-adrenergic receptor agonists, such as salbutamol, terbutaline, salmeterol, carmoterol, milveterol, and especially formoterol or indacaterol. Co-therapeutic antihistamine substances include cetirizine hydrochloride, clemastine fumarate, promethazine, loratadine, desloratadine diphenhydramine, and fexofenadine hydrochloride.

[0240] The compounds of the present invention can also be used in combination with one or more compounds that can be used to treat thrombolytic diseases, heart disease, stroke, etc. Such compounds include, but are not limited to: aspirin, streptokinase, tissue plasminogen activator, urokinase, anticoagulants, and antiplatelet drugs (e.g., PLAVIX). TM Clopidogrel bisulfate), statins (e.g., LIPITOR) TM Or atorvastatin calcium, ZOCOR TM (Simvastatin), CRESTOR TM (Rosuvastatin, etc.), beta-blockers (e.g., atenolol), NORVASC TM (amlodipine besylate) and ACE inhibitors (e.g. lisinopril).

[0241] The compounds of this invention can also be used in combination with one or more compounds that can be used to treat hypertension. Such compounds include, but are not limited to, ACE inhibitors, lipid-lowering agents such as statins, and lipid receptors. TM (Atorvastatin calcium) and calcium channel blockers (e.g., NORVASC) TM (Amlodipine besylate)

[0242] The compounds of the present invention can also be used in combination with one or more compounds selected from fibrates, β-blockers, NEPI inhibitors, angiotensin-2 receptor antagonists and platelet aggregation inhibitors.

[0243] The compounds of the present invention can also be used in combination with one or more compounds suitable for treating inflammatory diseases, including rheumatoid arthritis. Such compounds may be selected from: TNFα inhibitors such as anti-TNF-α monoclonal antibodies (e.g., REMICADE™, CDP-870) and D2E7 (HUMIRA™), as well as TNF receptor immunoglobulin fusion molecules (e.g., ENBREL™), IL-1 inhibitors, receptor antagonists, or soluble IL-1Rα (e.g., KINERET or ICE inhibitors), nonsteroidal anti-inflammatory drugs (NSAIDs), piroxicam, diclofenac, naproxen, flurbiprofen, fenoprofen, ketoprofen, and ibuprofen. ibuprofen), fenamic acid, mefenamic acid, indomethacin, sulindac, azapronone, pyrazolones, phenylbutazone, aspirin, COX-2 inhibitors (e.g., CELEBREX (celecoxib), PREXIGE (lumiracoxib)), metalloproteinase inhibitors (preferably selective MMP-13 inhibitors), p2x7 inhibitors, α2α inhibitors, NEUROTIN, pregabalin, low-dose methotrexate, leflunomide, hydroxychloroquine, d-penicillamine, auranoofin, or parenteral or oral gold.

[0244] The compounds of the present invention may also be used in combination with one or more compounds suitable for treating osteoarthritis. Such compounds may be selected from, but are not limited to, standard nonsteroidal anti-inflammatory drugs (hereinafter referred to as NSAIDs) such as piroxicam, diclofenac, propionic acid (e.g., naproxen), flurbiprofen, fenprofen, ketoprofen and ibuprofen, fenamic acid derivatives (e.g., mefenamic acid), indomethacin, sulindac, azaprozin, pyrazolone derivatives (e.g., phenylbutazone), salicylates (e.g., aspirin), COX-2 inhibitors (e.g., celecoxib, vardicoxib, rumicoxib and etorcoxib), analgesics and intra-articular therapies (e.g., corticosteroids and hyaluronic acid (e.g., hyalgan and syvisc))).

[0245] The compounds of this invention can also be used in combination with antiviral agents and / or antiseptic compounds. Such antiviral agents can be selected from, for example, Viracept, AZT, acyclovir, and famciclovir. Such antiseptic compounds can be selected from, for example, Valant.

[0246] The compounds of the present invention can also be used in combination with one or more pharmaceutical agents selected from the following: CNS agents such as antidepressants (sertraline), anti-Parkinson's disease drugs (e.g., deprenyl, L-DOPA, Requip, Mirapex; MAOB inhibitors (e.g., selegine and rasagiline); comP inhibitors (e.g., Tasmar); A-2 inhibitors; dopamine reuptake inhibitors; NMDA antagonists; nicotinic agonists; dopamine agonists; and neuronal nitric oxide synthase inhibitors). The compounds of the present invention can also be used in combination with one or more anti-Alzheimer's drugs. Such anti-Alzheimer's drugs may be selected from donepezil, tacrine, α2δ inhibitors, and NEURONTIN. TM Pregabalin, COX-2 inhibitors, propentofylline, or metrifonate.

[0247] The compounds of this invention can also be used in combination with osteoporosis agents and / or immunosuppressants. Such osteoporosis agents can be selected from EVISTA. TM (Raloxifene hydrochloride), droloxifene, lasofoxifene, or fosomax. Such immunosuppressants can be selected from FK-506 and rapamycin.

[0248] Generally, the compounds of the present invention are administered in a therapeutically effective amount via any acceptable method of administration for a similarly effective agent. The actual amount (i.e., the active ingredient) of the compounds of the present invention will depend on many factors, such as the severity of the disease to be treated, the age and relative health of the subject, the potency of the compound used, the route and form of administration, and other factors. The drug may be administered more than once a day, preferably once or twice a day. All these factors are within the skill of the attending clinician. The therapeutically effective amount of the compounds of the present invention can range from about 0.05 to about 50 mg / kg body weight / day of the recipient; preferably from about 0.1 to 25 mg / kg / day, more preferably from about 0.5 to 10 mg / kg / day. Thus, for administration to a 70 kg person, the dosage range is most preferably about 35 to 70 mg / day. In one embodiment, the therapeutically effective amount is 300 mg / day, optionally administered once daily.

[0249] Generally, the compounds of the present invention will be administered as pharmaceutical compositions via any of the following routes: oral, systemic (e.g., percutaneous, intranasal, or via suppository), or parenteral (e.g., intramuscular, intravenous, or subcutaneous). A preferred method of administration is oral administration using a convenient daily dosing regimen that can be adjusted according to the severity of the illness. The compositions may be in the form of tablets, pills, capsules, semi-solid dosage forms, powders, sustained-release formulations, solutions, suspensions, elixirs, aerosols, or any other suitable composition. Another preferred method of administration for compounds of formula (I) is inhalation. This is an effective method for delivering therapeutic agents directly to the respiratory tract.

[0250] The choice of formulation depends on a variety of factors, such as the route of administration and the bioavailability of the drug substance. For inhalation delivery, compounds can be formulated as liquid solutions, suspensions, aerosol propellants, or dry powders and loaded into suitable dispersers for administration. Several types of drug inhalation devices exist—nebulizers, metered-dose inhalers (MDIs), and dry powder inhalers (DPIs). Nebulizer devices generate a high-speed airflow that expels the therapeutic agent (formulated in liquid form) as a mist, which is carried into the patient's respiratory tract. MDIs are typically formulations packaged in compressed gas. Upon activation, the device expels a measured amount of the therapeutic agent via compressed gas, providing a reliable method for administering a set dose of the drug. DPIs disperse the therapeutic agent in a free-flowing powder form, which can be dispersed in the patient's inspiratory airflow during inhalation through the device. To achieve a free-flowing powder, the therapeutic agent is formulated with an excipient (e.g., lactose). The measured amount of the therapeutic agent is stored in capsule form and dispersed upon each activation. In some embodiments, the invention also relates to formulations in which the particle size of the compounds of the invention is 10 to 1000 nm, preferably 10 to 400 nm. Based on the principle that bioavailability can be improved by increasing surface area (i.e., reducing particle size), such pharmaceutical formulations have been developed specifically for drugs exhibiting poor bioavailability. For example, U.S. Patent No. 4,107,288 describes pharmaceutical formulations with particle sizes ranging from 10 to 1,000 nm, wherein the active substance is loaded on a cross-linked matrix of macromolecules. U.S. Patent No. 5,145,684 describes the production of pharmaceutical formulations in which the pharmaceutical substance is pulverized into nanoparticles (average particle size 400 nm) in the presence of a surface modifier and subsequently dispersed in a liquid medium to obtain a pharmaceutical formulation exhibiting significantly high bioavailability.

[0251] It should be understood that the dosage or amount of a compound and / or composition used alone or in combination with one or more active compounds to be applied depends on the individual circumstances and is conventionally suited to the individual environment to achieve optimal effect. Dosing and administration regimens are within the scope of those skilled in the art, and appropriate dosages depend on many factors within the knowledge scope of a general technical physician, veterinarian, or researcher (e.g., see Wells et al., eds., Pharmacotherapy Handbook, 2nd ed., Appleton and Lange, Stamford, Conn. (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, hardcover, Tarascon, Loma Linda, Calif. (2000)). For example, dosing and administration regimens may depend on the nature and severity of the condition to be treated, and also on the sex, age, weight, and individual responsiveness of the person or animal to be treated, the potency and duration of the action of the compound used, whether the treatment is acute or chronic or prophylactic, and / or whether other active compounds are administered in addition to the therapeutic molecule.

[0252] Therefore, the dosage of a compound or composition will vary depending on a variety of factors, including but not limited to: the activity, biological and pharmacokinetic properties and / or side effects of the compound used; the subject's age, weight, general health condition, sex, and diet; the time of administration, route of administration, excretion rate, and any drug combinations, if applicable; the effect the practitioner expects the compound to have in the subject; and the properties of the compound administered (e.g., bioavailability, stability, potency, toxicity, etc.). Such a suitable dosage can be determined as is known in the art. When one or more of the compounds of the present invention are administered to a human, for example, a physician may initially prescribe a relatively low dose, subsequently increasing the dose until an appropriate response is obtained.

[0253] There are no specific limitations on the dosage of each compound used in the compositions provided herein. Exemplary dosages include milligrams or micrograms of the compound per kilogram of object or sample weight (e.g., about 50 micrograms / kg to about 500 milligrams / kg, about 1 milligram / kg to about 100 milligrams / kg, about 1 milligram / kg to about 50 milligrams / kg, about 1 milligram / kg to about 10 milligrams / kg, or about 3 milligrams / kg to about 5 milligrams / kg). Other exemplary dosages include the following: about 5 to about 500 mg, about 25 to about 300 mg, about 25 to about 200 mg, about 50 to about 150 mg, or about 50, about 100, about 150 mg, about 200 mg, about 250 mg, or about 500 mg, and for example, once daily or twice daily, or lower or higher amounts.

[0254] In some embodiments, the adult dosage range is typically from 0.005 mg to 10 g / day orally. Tablets or other forms of presentation provided in discrete units can conveniently contain amounts of the compounds of the invention effective at such dosages or as multiples of such dosages, for example, containing 5 mg to 500 mg, typically about 10 mg to 200 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg units. Dosage units (e.g., oral dosage units) may include, for example, 1 to 30 mg, 1 to 40 mg, 1 to 100 mg, 1 to 300 mg, 1 to 500 mg, 2 to 500 mg, 3 to 100 mg, 5 to 20 mg, 5 to 100 mg (e.g., 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg). The compounds described herein (17 mg, 18 mg, 19 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg).

[0255] In some embodiments, the dosage range for oral administration is typically from about 0.001 mg to about 2000 mg of the compound per kg body weight. In some embodiments, the oral dose is from 0.01 mg / kg body weight to 100 mg / kg body weight, from 0.1 mg / kg body weight to 50 mg / kg body weight, from 0.5 mg / kg body weight to 20 mg / kg body weight, or from 1 mg / kg body weight to 10 mg / kg body weight. In some embodiments, the oral dose is 5 mg of the compound per kg body weight.

[0256] In other embodiments, the dosage is from about 10 mg to about 1000 mg, including all ranges and subranges therebetween, such as about 10 mg to about 900 mg, about 10 mg to about 800 mg, about 10 mg to about 700 mg, about 10 mg to about 600 mg, about 10 mg to about 500 mg, about 10 mg to about 400 mg, about 10 mg to about 300 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 150 mg, about 10 mg to about 100 mg, about 10 mg to about 50 mg, about 50 mg to about 900 mg, about 50 mg to about 800 mg, about 50 to about 700 mg, about 50 mg to about 600 mg, about 50 mg to about 500 mg, about 50 mg to about 400 mg, about 50 mg to about 300 mg, about 50 mg to about 250 mg, about 50 mg to about 200 mg, about 50 mg to about 150 mg, about 50 mg to about 100 mg, about 100 mg to about 900 mg, about 100 mg to about 800 mg, about 100 mg to about 700 mg, about 100 mg to about 600 mg, about 100 mg to about 500 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, About 100mg to about 200mg, about 100mg to about 150mg, about 150mg to about 200mg, about 150mg to about 250mg, about 150mg to about 300mg, about 150mg to about 400mg, about 150mg to about 500mg, about 200mg to about 900mg, about 200mg to about 800mg, about 200mg to about 700mg, about 200mg to about 500mg, about 200mg to about 400mg, about 200mg to about 300mg, about 200mg to about 250mg, about 300mg to about 900mg, about 300mg to about 800mg, about 300mg to about 800mg, about 30 0 to about 700 mg, about 300 to about 600 mg, about 300 mg to about 500 mg, about 300 mg to about 400 mg, about 400 mg to about 900 mg, about 400 mg to about 800 mg, about 400 to about 700 mg, about 400 to about 600 mg, about 400 mg to about 500 mg, about 500 mg to about 900 mg, about 500 mg to about 800 mg, about 500 to about 700 mg, about 500 to about 600 mg, about 100 mg to about 500 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, or about 100 mg to about 250 mg. In one embodiment, the range is about 150 mg to about 400 mg.

[0257] In some other embodiments, the dosage is 10 mg, 25 mg, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or 1000 mg.

[0258] In one implementation, the dosage is 300 mg per day, for example, 300 mg once a day, optionally with food.

[0259] Example

[0260] The invention will be more readily understood by referring to the following embodiments, which are provided to illustrate the invention and should not be construed as limiting its scope in any way.

[0261] Unless otherwise defined or the context clearly requires, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. It should be understood that any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this invention.

[0262] Example 1: N-(4-methyl-5-(2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridyl)-2-thiazolyl)aminocarbonyl-L-proline- 18 O-amide (compound 1, or apeliximab-) 18 Preparation of O1).

[0263]

[0264] 1-(tert-Butoxycarbonyl)-2-cyanopyrrolidine (100 mg, 0.51 mmol), palladium acetate (12 mg, 0.051 mmol), and 2,2'-bipyridine (8 mg, 0.051 mmol) were mixed with... 18 O-water (98%) 18 O- enrichment, 0.5 mL; also known as H2 18O) Mix in a sealed tube. Stir the mixture in the sealed tube at 60°C under a nitrogen atmosphere for 24 hours. Cool the reaction mixture to room temperature and concentrate under vacuum. Purify the residue by rapid column chromatography (gradient eluent DCM:MeOH at 100:0 to 50:1) to give N-Boc-L-proline- 18 O-amide (48mg; 18 O-enrichment, 97.3%. Towards N-Boc-L-proline- 18 A solution of O-amide (48 mg, 0.22 mmol) in DCM (0.5 mL) was added with 4 M HCl / dioxane (0.5 mL). The mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under vacuum to obtain L-proline- 18 O-Amide hydrochloride (35 mg). L-proline- 18 O-amide hydrochloride (33 mg, 0.22 mmol) was added to a solution of N-(4-methyl-5-(2-(2,2,2-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)-1H-imidazol-1-carboxamide (79 mg, 0.2 mmol) in DMF (1 mL), followed by the addition of triethylamine (80 mg, 0.8 mmol). The reaction mixture was stirred at 30 °C for 16 hours. DMF was removed under reduced pressure. The residue was purified by rapid column chromatography (eluent DCM:MeOH 100:0 to 30:1) to give the title compound (1, 70 mg, 81.9% yield). 18 O-enrichment (97.3%): 1 H NMR (500MHz, DMSO-d6) δppm10.96

[0265] (s, 1H), 8.60 (d, J=5.0Hz, 1H), 7.55 (s, 1H), 7.41 (s, 2H), 6.97 (s, 1H), 4.26 (s, 0.4 8H), 3.60(s, 1H), 3.46(s, 1H), 2.42(s, 4H), 2.09(s, 1H), 1.87(s, 3H), 1.61(s, 6H); 13 CNMR (125MHz, DMSO-d6) δppm174.36, 159.87, 158.99, 152.99, 149.58, 145.69, 141.44, 12 9.94, 127.69, 121.80, 120.78, 60.36, 55.40, 46.92, 46.71, 30.43, 21.93, 16.89.m / z (ESI + )4437, m / z(ESI - )441.5.

[0266] Example 2: N-(4-methyl-5-(2-(2,2,2c-trifluoro-1,1-dimethylethyl)-4-pyridyl)-2-thiazolyl)aminocarbonyl-L-proline- 17 O-amide (compound 2, or apeliximab-) 17 Preparation of O1).

[0267]

[0268] From O in the same manner as described for compound 1 17 -water (H2) 17 The preparation of this compound began with O).

[0269] Example 3: N-(4-methyl-5-(2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridyl)-2-thiazolyl)aminocarbonyl-L-proline- 13 C l -amide (compound 4, or apeliximab-) 13 Preparation of C1).

[0270]

[0271] From proline- in the same manner as described above 13 The compound was prepared by starting with C1-amide hydrochloride, yielding compound 4.

[0272] Example 4: N-(4-methyl-5-(2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridyl)-2-thiazolyl)aminocarbonyl-L-prolylamide- 15 N (compound 5, or apeliximab-) 15 Preparation of N).

[0273] L-proline (5 g, 43.4 mmol, 1.0 equivalent), triethylamine (6.6 g, 65.2 mmol, 1.5 equivalent), and (Boc)₂O (9.5 g, 43.5 mmol, 1 equivalent) were added to MeOH (50 mL); the mixture was stirred at 50 °C for 2 hours. The solvent was then removed under reduced pressure, and the residue was purified by rapid column chromatography (eluent: MeOH / DCM, 1 / 50) to give N-Boc-L-proline (6.0 g, 64.2%). Pyridine (0.36 mL, 4.20 mmol, 0.6 equivalent) was added to a solution of N-Boc-L-proline (1.5 g, 6.97 mmol, 1.0 equivalent) in 1,4-dioxane (27 mL). 15N-Ammonium sulfate (1.19 g, 8.87 mmol, 1.3 equivalents), (Boc)₂O (1.98 g, 8.87 mmol, 1.3 equivalents). The mixture was stirred overnight at 25 °C. The solvent was removed under reduced pressure, and the residue was purified by rapid column chromatography (eluent: DCM / MeOH = 100 / 0 to 50 / 1) to give N-Boc-L-proline- 15 N-amide (0.29 g, enriched 99.4%). To N-Boc-L-proline- 15 A solution of N-amide (0.29 g, 1.347 mmol, 1.0 equivalent) in DCM (3 mL) was added to dioxane in 4 M HCl (1.4 mL, 5.6 mmol, 4.16 equivalent). The mixture was stirred at room temperature for 0.5 h. The precipitate was filtered and dried under vacuum to give L-proline- 15 N-Amide hydrochloride (0.16 g, enriched 99.4%). CDI (0.4 g, 2.46 mmol) was added to a solution of 5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl-2-amine (0.5 g, 1.66 mmol) in DCM (25 mL), and the mixture was stirred at 40 °C for 4 hours. The reaction mixture was cooled to room temperature. The precipitate was filtered and dried under vacuum to give N-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)-1H-imidazol-1-carboxamide (0.5 g, 76.9%). L-proline- 15 N-amide hydrochloride (0.16 g, 1.06 mmol, 1.05 equivalence) was added to a solution of N-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)-1H-imidazol-1-carboxamide (400 mg, 1.01 mmol, 1.0 equivalence) in pyridine (5 mL), followed by the addition of DMAP (3 mg, 0.016 mmol, 0.023 equivalence). The reaction mixture was stirred overnight at 25 °C. The solvent was removed under reduced pressure, and the residue was purified by rapid column chromatography (eluent: DCM / MeOH = 100 / 0 to 30 / 1) to give the title compound (240 mg, 15N- enriched 99.4%, yield 53.7%): ¹H NMR (500MHz, DMSO-d6) δppm 1.64 (s, 6H), 1.90 (s, 3H), 2.12 (s, 1H), 2.44 (s, 3H), 3.50 (s, 1H), 3.63 (s, 1H), 4.31 (s, 1H), 6.99 (m, 1H), 7.42 (m, 2H), 7.58 (s, 1H), 8.63 (s, 1H), 10.96 (s, 1H); ¹³C NMR (125MHz, DMSO-d6) δppm16.78,21.91,24.39,30.41,46.72,46.92,60.38,120.68,12 1.71,125.39,127.65,129.90,132.15,141.42,149.53,159.00,174.21,174.32; m / z(ESI - 440.6, (ESI) + )442.8.

[0274] Example 5: Pharmacokinetic study of compound 1 in Sprague Dawley (SD) rats.

[0275] (a) SD rats were randomly divided into groups of 6 (n=6). Compound 1 (apelis-) was administered to each group. 18 Compound 1 (O1) was mixed with apegliflozin and adjusted to a 1:1 molar ratio in the administration solution (1.25 mg / mL); the administration solution was then administered to animals orally at a dose level of 2.5 mg / kg. Blood samples were collected at predetermined time points 0.167, 0.5, 1, 2, 3, 4, 6, 8, and 24 hours (h) following drug administration. Blood samples were converted to plasma samples using standard techniques, and the latter samples were analyzed to determine the concentrations of compound 1 and apegliflozin. Data are shown in Table 2a and Figure 1a As shown in [the image]. Figure 1a In the diagram, the lines marked with -○- and --Δ-- represent the concentrations of apelips at the same molar dose. 18 Following oral administration of O1 and apelistat to animals, apelistat- 18 Plasma concentrations of O1 and apeliximab.

[0276] Table 2a. PK parameters in SD rats (dose: 2.5 mg / kg).

[0277] <![CDATA[AUC 0至t ]]> ug / L*hour 28719 26405 <![CDATA[AUC o至∞ ]]> ug / L*hour 28798 26476 <![CDATA[T 最大 ]]> Hour 3 3 Vz / F L / kg 1.655 1.798 CLz / F L / hour / kg 0.434 0.472 <![CDATA[C 最大 ]]> ug / L 2390 2297

[0278] (b) Similarly, compound 1 (apelis- 18Compound 1 and apegliflozin were administered orally to animals (SD rats, n=4) at low doses (0.322 mg / kg). Blood samples were collected at pre-specified time points of 0.167, 0.5, 1, 2, 4, 6, 8, 10, and 24 hours (h) following drug administration. Blood samples were converted to plasma samples using standard techniques, and the latter were analyzed to determine the concentrations of compound 1 and apegliflozin. At the 24-hour time point, the concentrations of both compounds were below the limit of quantification (BLQ); therefore, data from this time point were discarded for data analysis purposes. Data are shown in Table 2b and Figure 1b As shown in [the image]. Figure 1b In the diagram, the solid and dashed lines represent Apelles- 18 Plasma concentrations of O1 and apeliximab.

[0279] Table 2b. PK parameters in SD rats (dose: 0.322 mg / kg).

[0280] <![CDATA[AUC 0至t ]]> ug / L*hour 117.8 75.2 <![CDATA[AUC 0至∞ ]]> ug / L*hour 131.7 80.3

[0281] Vz / F L / kg 11.4 13.1 CLz / F L / hour / kg 2.5 4.1 <![CDATA[C 最大 ]]> ug / L 32.6 28.1

[0282] The results showed that, after administration under the same conditions, apeliximab- 18 Plasma concentrations of O1 were higher than those of apegliflozin. This effect was observed at both tested doses, with the best results seen at the lower dose.

[0283] Example 6: Pharmacokinetic study of compound 1 in ICR mice.

[0284] Forty-eight ICR mice were randomly divided into 6 groups (n=8). Compound 1 (apelis-) was administered to each group. 18 Compound 1 (O1) was mixed with apegliflozin and adjusted to a 1:1 molar ratio in the administration solution (1.25 mg / mL); the administration solution was then administered to animals orally at a dose level of 2.5 mg / kg. Following administration, blood samples were collected at time points of 0.5, 1, 2, 4, 8, and 12 hours (for each time point, a group of 8 animals). Blood samples were converted to plasma samples using standard techniques, and the latter samples were analyzed to determine the concentrations of compound 1 and apegliflozin. Data are shown in Table 3 and... Figure 2 As shown in [the image]. Figure 2 In the diagram, the lines marked with -○- and --Δ-- represent the concentrations of the same dose of apelips. 18 Following oral administration of O1 and apelistat to animals, apelistat- 18Plasma concentrations of O1 and apegliflozin. Results showed that, after administration under the same conditions, apegliflozin- 18 The plasma concentration of O1 was higher than that of apegliflozin.

[0285] Table 3. PK parameters in ICR mice.

[0286] AUC(0 to t) ug / L*hour 27845 26212 AUC (0 to ∞) ug / L*hour 32554 30597 <![CDATA[T 最大 ]]> Hour 2 2 Vz / F L / kg 2.236 2.37 CLz / F L / hour / kg 0.384 0.409 <![CDATA[C 最大 ]]> ug / L 4519 4208

[0287] Example 7: M4 and M4- in liver S9 culture medium 18 The generation of O1.

[0288] The major metabolite of apelips is M4, an acidic form of a proline derivative, obtained by enzymatic and / or chemical hydrolysis of the amide bond, as shown below:

[0289]

[0290] Dilute equimolar concentrations of apelips and apelips- with pre-warmed (37°C, water bath) 0.1M Tris-acetate buffer. 18 O1 stock solution (50 μL); and the concentration of each compound in this working solution is 4.5 μM. Add the above working solution (50 μL) and liver S9 solution (50 μL) to each incubation well and mix thoroughly. Pre-incubate the sample for 5 minutes, then add β-NADPH dosing solution (100 μL) to each well. Incubate the sample plate at 37 °C and analyze in triplicate at time points of 0, 15, 30, 60, and 240 minutes. Sample analysis: At the preset time point, add stop solution (800 μL, acetonitrile) to each well; mix the sample thoroughly (vortex) and centrifuge at 12000 rpm for 5 minutes; transfer the supernatant to the analytical sample vial and then analyze M4 and M4- by LC-MS / MS. 18 O1 was analyzed. Quantitative analysis of the data: In separate experiments under the same analytical conditions, (1) M4 and M4- were determined. 18 O1 exhibited the same response across the experimental concentration range; and (2) apelis was incubated under the same conditions. 18 No M4 was observed after O1, indicating that no cross-generation of M4 and M4- occurred from the incubation of compound 1 and apeliximab. 18 O. Liver S9 used in the experiment: (a) monkey liver S9, (b) combined human S9, (c) miniature pig liver S9, and (d) male Wistar rat liver S9. Experimental results are summarized in Figures 3a to 3d middle.

[0291] The results showed that, consistent with the observed increase in plasma concentrations of compound 1 compared to apegliflozin, the plasma concentrations derived from apegliflozin... 18O1 metabolite M4- 18 O1 plasma concentration and from 16 The plasma concentration of compound O1 (apelis) was lower than that of M4. Furthermore, the results determined that, similar to apelis, the major metabolite of compound 1 is a hydrolysis product of the right-terminal proline amide group, namely an N-substituted proline- 18 O1 or M4- 18 O1. Additionally, the free carboxyl group of proline derivatives contains only one oxygen atom. 18 O (i.e., M4-) 18 O1), indicating that in apelis- 18 No oxygen-18 atom was lost during the metabolism of O1. In summary, the results indicate that the metabolism of compound 1 is reduced compared to the metabolism of apegliflozin.

[0292] Example 8: Pharmacokinetic study of compound 1 in beagle dogs.

[0293] Male and female beagle dogs (n=4; 2 males and 2 females) were used in the study. A single oral dose of apelips (2.5 mg / kg) and compound 1 were administered co-administered in DMSO and a 0.5% carboxymethyl cellulose sodium (CMC-Na) aqueous solution (5:95, v / v). 18 O1-Apeliximab (2.5 mg / kg) (total dose 5 mg / kg). Blood samples were collected at 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 10, 12 and 24 hours for analysis of apeleliximab and... 18 O1-Apeliximab. Data are in Table 4 and... Figure 4 As shown in [the image]. Figure 4 In the diagram, the lines marked with --○-- and -Δ- represent the concentrations of the same dose of apelips- 18 Following oral administration of O1 and apelistat to animals, apelistat- 18 Plasma concentrations of O1 and apegliflozin. Results showed that, after administration under the same conditions, apegliflozin- 18 The plasma concentration of O1 was higher than that of apegliflozin.

[0294] Table 4. PK parameters in Beagles.

[0295] <![CDATA[AUC (0至t) ]]> ug / L*hour 6682 6115 <![CDATA[t 1 / 2 ]]> Hour 4.5 5.4 <![CDATA[T 最大 ]]> Hour 1.1 1.2 Vz / F L / kg 2.4 3.0 CLz / F L / hour / kg 0.38 0.41 <![CDATA[C 最大 ]]> ug / L 1468 931

[0296] In summary, the studies in the above embodiments show that the plasma concentration of compound 1 is higher than that of apeliximab, which is consistent with the reduced metabolism of proline amide groups and the increased bioavailability of compound 1.

[0297] Nirapanib, another isotope-enriched compound, in SD rats and beagle dogs. 18 O underwent similar research. However, this isotope-enriched compound is similar to nirapanib- 16 Compared to O, no improvement in plasma concentration or bioavailability was observed (data not shown).

[0298] Although the invention has been described in detail with reference to embodiments thereof, these embodiments are provided for illustrative purposes and not for limiting the invention. Other embodiments may be made that employ the principles of the invention and fall within the spirit and scope defined by the appended claims.

[0299] All documents and references cited in this article are incorporated herein by reference in their entirety.

Claims

1. Compounds enriched in formula II isotopes, or their pharmaceutically usable salts: (II), in, It has one or more selected from 18 O and 17 The stable heavy isotope enrichment of the amide functional group of O; and R 1 and R 2 It is independently hydrogen or a protecting group selected from acyl, carbonyl, thiocarbonyl and carbamoyl.

2. The isotope-enriching compound of claim 1, wherein the heavy isotope-enriching amide functional group is enriched with 18 O、 17 O or 18 O and 17 A mixture of O.

3. The isotope enrichment compound according to claim 1 or 2, wherein R 1 and R 2 It is hydrogen.

4. The isotope-enriching compound of claim 2, wherein the heavy isotope-enriching amide functional group is enriched with 18 O.

5. An isotope-enriched compound comprising a heavy isotope-enriched amide functional group, wherein the isotope-enriched compound is the following or a pharmaceutically acceptable salt thereof: ， Where O* represents 18 O and 17 Enrichment of O.

6. The isotope enrichment compound according to any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 5% or more.

7. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 10% or more.

8. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 20% or more.

9. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 50% or more.

10. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 75% or more.

11. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 90% or more.

12. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 95% or more.

13. The isotope enrichment compound of any one of claims 1 to 5, wherein the heavy isotope enrichment level in the isotope enrichment compound is about 98% or more.

14. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 90% or higher, optionally determined using mass spectrometry.

15. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 91% or higher, optionally determined using mass spectrometry.

16. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 92% or higher, optionally determined using mass spectrometry.

17. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 93% or higher, optionally determined using mass spectrometry.

18. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 94% or higher, optionally determined using mass spectrometry.

19. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 95% or higher, optionally determined using mass spectrometry.

20. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 96% or higher, optionally determined using mass spectrometry.

21. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 97% or higher, optionally determined using mass spectrometry.

22. The isotope enrichment compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope enrichment compound is about 98% or higher, optionally determined using mass spectrometry.

23. The isotope-enriched compound of any one of claims 1 to 5, wherein the isotope enrichment level in the isotope-enriched compound is about 99% or higher, optionally determined using mass spectrometry.

24. A pharmaceutical composition comprising the isotope-enriched compound of any one of claims 1 to 23 and a pharmaceutically acceptable carrier.

25. The pharmaceutical composition of claim 24, wherein the composition is in a form suitable for oral administration.

26. The pharmaceutical composition of claim 24 or 25, wherein the composition is in the form of: hard-shell gelatin capsules, soft-shell gelatin capsules, flat capsules, pills, tablets, lozenges, powders, granules, solutions, aqueous liquid suspensions, non-aqueous liquid suspensions, oil-in-water liquid emulsions, water-in-oil liquid emulsions, elixirs, or syrups.

27. The pharmaceutical composition of claim 24 or 25, wherein the composition is in the form of granules, soft tablets or sugar-coated pills.

28. The pharmaceutical composition of claim 24 or 25, wherein the composition is enteric-coated or formulated for controlled release.

29. Use of the isotope-enriched compound of any one of claims 1 to 23 or the pharmaceutical composition of any one of claims 24 to 28 in the preparation of a medicament for treating breast cancer in a subject.

30. The use as claimed in claim 29, wherein the object is a mammal.

31. The use as claimed in claim 30, wherein the mammal is a human.

32. The use according to claim 31, wherein the breast cancer is hormone receptor positive (HR). + And human epidermal growth factor receptor negative (HER2) - )of.

33. The use according to claim 31 or 32, wherein the breast cancer is PIKCA mutated.

34. The use as described in claim 29 or 30, wherein the use further comprises using a therapeutically effective amount of fulvestrant.

35. The use according to claim 34, wherein the fulvestrant is administered simultaneously with the isotope-enriched compound of any one of claims 1 to 23 or the pharmaceutical composition of any one of claims 24 to 28.

36. The use according to claim 34, wherein the fulvestrant is administered sequentially with the isotope enrichment compound of any one of claims 1 to 23 or the pharmaceutical composition of any one of claims 24 to 28.

37. The use according to any one of claims 29 to 32, wherein the isotope-enriched compound or the pharmaceutical composition is administered orally.

38. The use according to claim 37, wherein the pharmaceutical composition is in tablet form.

39. The use according to any one of claims 29 to 32, wherein the therapeutically effective amount of the isotope-enriched compound is about 50 mg to about 500 mg.

40. The use according to claim 39, wherein the therapeutically effective dose is 300 mg administered once daily.

41. A medicine box comprising the isotope enrichment compound of any one of claims 1 to 23 or the pharmaceutical composition of any one of claims 24 to 28, and instructions for use thereof.

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