CN117942338A

CN117942338A - Pharmaceutical use of isotopically enriched compounds in PROS

Info

Publication number: CN117942338A
Application number: CN202211294381.XA
Authority: CN
Inventors: 吕佳声; 叶祥胜; 周天伦
Original assignee: Junshi Runjia Shanghai Pharmaceutical Technology Co ltd
Current assignee: Junshi Runjia Shanghai Pharmaceutical Technology Co ltd
Priority date: 2022-10-21
Filing date: 2022-10-21
Publication date: 2024-04-30

Abstract

The present invention provides the use of a compound of formula (I) for the manufacture of a medicament for the treatment of a PI 3K-related disease or disorder, in particular PIK3 CA-related overgrowth syndrome group (PROS), in particular including congenital lipid overgrowth, vascular malformations, epidermal nevi and spinal/skeletal abnormalities and/or scoliosis syndrome (cage), and the like.

Description

Pharmaceutical use of isotopically enriched compounds in PROS

Technical Field

The invention relates to the use of a heavy isotopically enriched compound in a medicament for the treatment of overgrowth diseases associated with PI3K, in particular the PIK3 CA-associated overgrowth syndrome group (PROS).

Background

Activation of phosphoinositide kinase (Phosphatidylinositol-3-kinase, PI 3K) leads to various conditions in humans. PIK3CA belongs to the PI3K/AKT pathway, which is located on chromosome 3 and is mainly used to encode the p110a protein, whereas the p110a protein is one of the catalytic subunits of PI3K enzymes. Somatic activation mutations in the PI3K/AKT/mTOR pathway are the basis for excessive growth of the catabolic segment. Historically, clinical diagnostics associated with PIK3CA activating mutations include fibrofatty overgrowth (Fibroadipose hyperplasia or Overgrowth, FAO), hyperproliferative adiposity (HEMIHYPERPLA-sia Multiple Lipomatosis, HHML), congenital, lipidic, overgrowth, vascular malformations, epidermal nevi and spinal/skeletal abnormalities and/or scoliosis (Congenital Lipomatous Overgrowth、Vascular Malformations、Epidermal Nevi、Scolio-sis/Skeletal and Spinal syndrome,CLOVES), macrodeformity (macrodactyly), fibrosteatosis (Fibroadipose Infiltrating Lipomatosis), megabrain-capillary malformations (MEGALENCEPHALY-CAPILLARY MALFORMATION, MCAP), and dysplastic megabrain (DYSPLASTIC MEGALENCEPHALY, DMEG), among others. Such clinical symptoms associated with PIK3CA activating mutations are collectively referred to as PROS, i.e., PIK3CA-Related Over-growth Spectrum(Kim M.American Journal of Medical Genetics,Part A(2015),167(2),287-295), may also be referred to as PIK3CA-related overgrowth syndromes.

Disclosure of Invention

It is an object of the present invention to provide the use of an isotopically enriched compound in a medicament for the treatment or prophylaxis of a PROS condition associated with a PIK3CA activating mutation.

Alpelisib as PI3K inhibitors, are amide containing compounds having the following structure:

Which has an amide function, a prolinamide group derived from a proline moiety shown at the right end of the molecule.

In an embodiment, the present application provides the use of a compound of formula (I) or a pharmaceutically acceptable salt, ester, hydrate, chelate and/or solvate thereof, in a medicament for the treatment of PROS:

Wherein the amide bond A heavy isotopically enriched amide bond comprising at least one isotope of ¹⁸O、¹⁷O、¹³ C and ¹⁵ N;

r ¹ and R ² are independently hydrogen or a protecting group.

In some embodiments, at least one of R ¹ and R ² of the compound of formula (I) is not hydrogen. In some embodiments, the compound of formula (I) is present in the form of a prodrug.

In some embodiments, in the compound of formula (I), the heavy isotopically enriched amide bond orIs monoisotopically enriched, e.g., ¹⁸ O-enriched, ¹⁷ O-enriched, ¹³ C-enriched, or ¹⁵ N-enriched; and R ¹ and R ² have the same definition as above.

In another embodiment, in the compound of formula (I), the heavy isotopically enriched amide bond orTwo or more heavy isotopes of one or more elements are enriched, for example ¹⁷ O-and ¹⁸ O-enriched, ¹⁸ O-and ¹³ C-enriched, ¹⁸ O-and ¹⁵ N-enriched or ¹³ C-and ¹⁵ N-enriched.

In some embodiments, the level of isotopic enrichment in the compound is about 5% or more, about 10% or more, about 20% or more, about 50% or more, about 75% or more, about 90% or more, about 95% or more, about 97% or more, about 98% or more, about 99% or more, or 100%.

In some embodiments, the level of isotopic enrichment in the compound is 5% or more, 10% or more, 20% or more, 50% or more, 75% or more, 90% or more, 95% or more, 97% or more, 98% or more, 99% or more, or 100%.

In particular embodiments, compounds of table 1 or pharmaceutically acceptable salts, esters, hydrates, chelates, and solvates thereof are provided.

Table 1: examples of the Compounds of the invention

Wherein, the chemical bond-c=o is represented by O and/or ¹⁷ O and/or ¹⁸ O, and compound 3 may represent a mixture, i.e. compound 3 may be a mixture of a compound with O being O ¹⁷ and a compound with O being O ¹⁸. Specifically, the ratio of the compound having O of ¹⁷ may be about 5% or more, about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 75% or more, about 90% or more, about 95% or more, about 97% or more, about 98% or more, or about 99% or more, or 100%.

The present invention also provides a pharmaceutical composition comprising a compound as described herein, or a pharmaceutically acceptable salt, ester, hydrate, chelate or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, any of the compounds described above may be used to treat congenital lipid overgrowth, vascular malformations, superficial nevi, and spinal/skeletal abnormalities, and/or scoliosis syndrome (CLOVES).

In some embodiments, any of the compounds described above may be used to treat fibrofatty overgrowth (FAO), hyperproliferative adiposity (HHML), datagram, fibrolipoinvasive lipoma, megabrain-capillary deformity (MCAP), or Dysplastic Megabrain (DMEG).

In some embodiments, any of the compounds described above may be used to treat Klippel-Trenaunay syndrome.

In one embodiment, the compounds (e.g., compounds of formula I, or compounds of table 1) and/or pharmaceutical compositions of the invention are useful for modulating a metabolic pathway of a drug or prodrug, reducing the metabolism of a drug or prodrug, modulating the pharmacokinetic properties of a drug or prodrug, and/or improving or increasing the therapeutic effect of a drug or prodrug in a subject, as compared to using a compound having only a naturally abundant isotopic atom (i.e., a compound that is not isotopically enriched). In some embodiments, the compounds and pharmaceutical compositions provided herein are used to reduce therapeutic toxicity and/or side effects of the compounds, increase drug tolerance, and/or improve or increase the therapeutic or prophylactic effect of the compounds in a subject, as compared to using compounds having only isotopic atoms in natural abundance (i.e., compounds that are not isotopically enriched). In some embodiments, the compounds and pharmaceutical compositions provided herein are used to improve the biodistribution of the compound and/or enhance the therapeutic and/or prophylactic effects of the compound in a subject, as compared to using a compound having only isotopically enriched atoms in natural abundance (i.e., a compound that is not isotopically enriched).

In one embodiment, a method of modulating the metabolic or pharmacokinetic properties of an amide bond containing compound in a subject is provided, comprising administering an isotopically enriched compound or pharmaceutical composition described herein to a subject, wherein the metabolic and pharmacokinetic properties of the amide bond containing compound of the invention are modulated compared to administration of the same compound having only isotopically enriched natural atoms (i.e., a compound that is not isotopically enriched). In some embodiments, methods of reducing metabolism of a compound, reducing therapeutic toxicity of a compound, reducing side effects of a compound, increasing tolerance of a compound, improving biodistribution of a compound, and/or increasing therapeutic or prophylactic effects of a compound in a subject are provided, the methods comprising administering an isotopically enriched compound or pharmaceutical composition of the invention to a subject, wherein metabolism of a compound is reduced, therapeutic toxicity is reduced, side effects are increased, tolerance is increased, biodistribution is improved, and/or therapeutic or prophylactic effects are increased as compared to administration of a compound having only a natural abundance isotopically atom (i.e., a compound that is not isotopically enriched).

In other general aspects, kits are provided that include one or more of the compounds or pharmaceutical compositions described herein. The kit may further include one or more additional therapeutic agents, and/or instructions, for example instructions for using the kit to treat a subject suffering from the same disease or disorder as the parent compound (the compound that is not isotopically enriched).

Drawings

For a better understanding of the invention, and to show more clearly how it may be carried into effect, reference will now be made, by way of example, to the accompanying drawings which show aspects and features in accordance with embodiments of the invention, and in which:

FIG. 1 shows drug concentration versus time curves for Alpelisib and Alpelisib- ¹⁸O₁ in plasma in SD rats orally administered Alpelisib and Alpelisib- ¹⁸O₁, respectively, at the same dose;

FIG. 2 shows drug concentration versus time curves in plasma Alpelisib and Alpelisib- ¹⁸O₁ in ICR mice orally administered Alpelisib and Alpelisib- ¹⁸O₁, respectively, at the same dose;

Figures 3a to 3d show the production of M4 and M4- ¹⁸O₁ after incubation Alpelisib and Alpelisib- ¹⁸O₁ in the medium of liver S9 of various species: (a) monkey liver S9, (b) mixed human liver S9, (c) mini pig liver S9, and (d) male Wistar rat liver S9. In each figure, M4: -x-; m4- ¹⁸ O: -o-.

Detailed Description

Definition of the definition

In order to provide a clear and consistent understanding of the terms used in the description of the present invention, some definitions are provided below. Furthermore, unless defined otherwise, 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 belongs.

When used in conjunction with the term "comprising" in the claims and/or the specification, the use of the word "a" or "an" may mean "one or more", "at least one" and "one or more" as well. Similarly, the word "another" may mean at least a second or a plurality.

As used herein, the term "PROS" refers to the PIK3CA related overgrowth syndrome population. It is a group of diseases such as fibrofatty overgrowth (FAO), megabrain-capillary Malformation (MCAP) syndrome, congenital lipoma of the torso, asymmetric overgrowth, lymphatic, capillary, venous and combined vascular malformations, superficial nevi, bone and spinal abnormalities (clear) syndrome and hyperproliferative adiposity (HHML) and Klippel-Trenaunay syndrome.

As used herein, the term "fibrous fat overgrowth (FAO)" refers to a syndrome characterized by the following major findings: segmental progressive overgrowth of subcutaneous, muscle and visceral fibrous lipid tissue with skeletal overgrowth (Lindhurst et al, 2012).

As used herein, the term "megabrain-capillary Malformation (MCAP) syndrome" refers to a syndrome characterized by the following major findings: (1) Megabrain disease (MEG) or hemilateral megabrain disease (HMEG) associated with hypotonia, epilepsy, and neurological findings of mild to severe mental disabilities; (2) Skin capillary deformities associated with focal or systemic somatic overgrowth (Mirzaa et al, 2013).

In a specific embodiment, the PROS disease is CLOVES. As used herein, the term "outside" refers to congenital, lipidic, overgrowth, vascular malformations, epidermal nevi, and spinal/skeletal abnormalities and/or scoliosis. This syndrome is characterized by a complex congenital overgrowth of the lipid tissue (often manifested as a trunk lipid tumor) and a combination of vascular and lymphatic vessel malformations.

As used herein, the term "hyperproliferative multiple adiposity (HHML)" refers to a condition characterized by asymmetric non-progressive overgrowth, multiple lipomas, and superficial vascular malformations (BG et al 2013).

In a specific embodiment, the PROS disease is Klippel-Trenaunay syndrome. As used herein, the term "Klippel-Trenaunay syndrome" refers to a rare congenital medical condition in which blood and/or lymphatic vessels are not properly formed.

Thus, a subject that may be diagnosed as exhibiting one of the diseases in PROS provides a method according to the invention.

As used herein, the term "subject" refers to any mammal, such as rodents, felines, canines, and primates. In particular, in the present invention, the subject is a human suffering from or susceptible to PROS disease. In a specific embodiment, the subject is a human suffering from or susceptible to CLOVES syndrome. In a specific embodiment, the subject is a human suffering from or susceptible to Klippel-Trenaunay syndrome.

As used herein, the term "naturally abundant oxygen" refers to a natural isotopically abundant oxygen atom, typically having an isotopic composition of oxygen atoms close to the earth's atmosphere: ¹⁶O,99.759％;¹⁷ O,0.037%; and ¹⁸ O,0.204%. However, it should be understood that naturally occurring compounds may have slight variations in the isotopic composition of the oxygen atoms.

The terms "heavy oxygen atom", "heavy oxygen isotope", "stabilized heavy oxygen", "^* O" and "O ^*" are used interchangeably herein and refer to a stabilized oxygen atom of ¹⁷ O and/or ¹⁸ O, excluding any radioactive and non-naturally occurring heavy isotopes. ¹⁷ O and ¹⁸ O occur naturally but at very low rates relative to the main isotope ¹⁶ O. The terms "heavy carbon isotope", "stable heavy carbon isotope", "carbon-13", "¹³ C" and "^* C" are used interchangeably herein to refer to the ¹³ C isotope. Similarly, the terms "diazonium isotope", "stable diazonium isotope", "nitrogen-15", "¹⁵ N" and "^* N" are used interchangeably herein to refer to the ¹⁵ N isotope.

As used herein, the terms "oxygen atom enriched with a stable heavy oxygen isotope", "stable heavy oxygen enriched", "^* O-enriched" and "O ^* -enriched" are used interchangeably herein to refer to heavy oxygen atoms that do not have their natural isotopic composition but rather have a higher stable heavy oxygen isotope than the naturally occurring isotopic composition. "heavy oxygen enriched", "stable heavy oxygen enriched", "^* O-enriched" and "O ^* -enriched" refer to compounds in which the oxygen atom at a particular site is enriched by a stable heavy oxygen isotope, in particular, by one or both of ¹⁷ O and ¹⁸ O.

Likewise, with respect to carbon or nitrogen atoms enriched with stable heavy atom isotopes, ¹³ C-enriched or ¹⁵ N-enriched means that the proportion of heavy isotopes ¹³ C or ¹⁵ N in carbon or nitrogen atoms, respectively, in compounds without natural isotopic composition is higher than in naturally occurring carbon or nitrogen, respectively.

Isotopic enrichment is a method of altering the relative abundance of isotopes of a given element, thereby producing a form of the element in which one particular isotope is enriched (i.e., increased) while the other isotopes are reduced or depleted. As used herein, a "heavy isotopically enriched" or "isotopically enriched" compound or derivative refers to a compound enriched (i.e., augmented) with a specific isotope, i.e., oxygen-17 or oxygen-18, or both, and/or carbon-13, and/or nitrogen-15, at a specific position of the compound or at the site of an amide bond.

For carbon and nitrogen, the mole fractions of the most abundant heavy isotopes ¹³ C and ¹⁵ N are 0.0107 and 0.00364, respectively, compared to the most abundant isotopes ¹⁴ N of the most abundant isotopes ¹² C and 0.99636, respectively, of 0.9893. Under normal conditions, the mole fractions of oxygen-18 (¹⁸ O) and oxygen-17 (¹⁷ O) are 0.00204 and 0.00037, respectively, relative to the most abundant isotope ¹⁶ O with a mole fraction of 0.99757.

As used herein, an "isotopically enriched" or "heavy isotopically enriched" compound or derivative has a content of isotopically enriched forms higher than its natural abundance. The degree of enrichment of isotopes varies depending on the natural abundance of a particular isotopic form. In some embodiments, the compound or the isotopic enrichment level of the element in the compound may be about 2 mole percent to about 100 mole percent (%), 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 isotopically enriched compounds of the present invention (e.g., compounds of formula I or compounds of table 1, etc.) have a degree of isotopic enrichment of about 5% or more, or about 10% or more. In another embodiment, the isotopically enriched compounds of the present invention (e.g., compounds of formula I and compounds described herein) have a degree of isotopic enrichment of about 20% or more, or about 50% or more. In yet another embodiment, the isotopically enriched compounds of the present invention (e.g., compounds of formula (I) and compounds described herein) have a degree of isotopic enrichment of about 75% or more, or about 90% or more. In yet another embodiment, the isotopically enriched levels in the isotopically enriched compounds of the present invention (e.g., compounds of formula I and compounds described herein) are about 95% or more, about 98% or more, or 100%. It will be appreciated that the isotopic enrichment level of a particular compound, or the particular oxygen isotope of a compound, will be chosen according to several properties of the compound, including chemical, pharmacokinetic and therapeutic properties, etc., to enhance the therapeutic or prophylactic effect, therapeutic biodistribution, bioavailability, metabolism, stability and/or pharmacokinetic properties of the compound.

As used herein, a "non-isotopically enriched" compound refers to a compound in which all atoms or elements in the compound are isotopes of natural abundance, i.e., all atoms or elements have the most abundant atomic mass in nature. The non-isotopically enriched compounds are quite different from isotopically enriched compounds in which one or more elements are enriched to one or more specific isotopic forms instead of the naturally abundant isotopes. Compounds that are isotopically enriched are not encompassed by the present compounds provided herein.

As used herein, the terms "compounds of the invention" and equivalent expressions refer to isotopically enriched compounds provided herein which are useful for at least one purpose of the present invention, for example, those encompassed by structural formulas such as formula I, as well as the specific compounds mentioned herein, and pharmaceutically acceptable salts, esters, chelates, hydrates and/or solvates thereof.

Without being limited by theory, the two heavy oxygen isotopes, ¹⁸ O and ¹⁷ O, have different isotopic effects on the cleavage of the amide bond, e.g., the rate of hydrolysis reaction. ¹⁸ O, having an atomic weight of 18 daltons, has an isotopic effect higher than ¹⁷ O (having an atomic weight of 17). Enrichment with ¹⁸ O or ¹⁷ O, or with a mixture of ¹⁸ O and ¹⁷ O in different proportions, may be selected to achieve the desired biological and pharmaceutical effects.

Likewise, heavy isotopes ¹³ C and/or ¹⁵ N may be selected for enrichment to achieve the desired biological and pharmaceutical effects of isotopically enriched compounds.

As described herein, isotopically enriched compounds can be isotopically enriched with one or more isotopes of a single element, or with isotopes of one or more elements.

As used herein, "drug" or "prodrug" or "parent compound" refers to a compound that has natural isotopic abundance at the amide site of interest. The present invention provides such compounds in a "heavy isotopically enriched" form, compositions and methods of preparation and use, thereby modulating or improving the pharmaceutical properties of the parent compound, and enhancing or modulating the efficacy of the treatment of a disease. In some embodiments, a "drug" or "prodrug" or "parent compound" refers to a compound having natural oxygen, carbon, and/or nitrogen abundance at each amide oxygen, carbon, and/or nitrogen position of interest; the present invention provides compounds, compositions, methods of preparation, and uses thereof, of the "heavy oxygen", "heavy carbon", and/or "diazonium" enriched forms of such compounds, thereby modulating or improving the pharmaceutical properties of the parent compound and enhancing or modulating the efficacy of the treatment of the disease.

As used herein, "organic moiety" or "organic fragment" refers to a group of atoms that are part of the entire molecule or structure of a compound. The moieties, amide linkages or functional groups, protecting groups or substituents, if present, are covalently linked together to form a molecule or structure of the compound.

As used herein, the term "protecting group" has a meaning well known in the art, in the present application, especially an amino protecting group, such as t-butoxycarbonyl, acetyl, (t-butyldimethylsilyl) oxy, benzyloxycarbonyl, p-toluenesulfonyl, and the like.

In one embodiment, when substitution is capable, the compounds of the present invention may be further substituted with one or more substituents. In some embodiments, the substituted form of the compound is a prodrug; in these embodiments, the substituents may be cleaved, or the compounds otherwise converted, to release the active ingredient or drug molecule from the prodrug form upon administration to a subject.

The term "administration" or "dosing" refers to the delivery of a compound to a subject, including all metering and drug delivery means known in the art.

As used herein, the word "comprising" (and any form of comprising, such as "comprising" and "comprises"), "having" (and any form of having, "having", "including" and "containing") is inclusive and open-ended and does not exclude additional unrecited elements or process steps.

The term "about" is used to indicate that the value includes errors in the instruments and methods used in determining the value.

The term "derivative" as used herein refers to a substance that is structurally similar to another compound but structurally different in some minor structures.

The present specification relates to a number of chemical terms and abbreviations used by those skilled in the art. However, for the sake of clarity and consistency, definitions of selected terms are provided.

As used herein, the term "substituted" or "having a substituent" means that the parent compound or moiety has at least one substituent group. The term "unsubstituted" or "without substituents" means that the parent compound or moiety has no substituents other than chemical saturation of the undefined valence with a hydrogen atom.

As used herein, "substituent" or "substituent group" refers to a moiety selected from the group consisting of halogen (F, cl, br, or I), hydroxy, mercapto, amino, nitro, carbonyl, carboxyl, alkyl, alkoxy, alkylamino, aryl, aryloxy, arylamino, acyl, sulfinyl, sulfonyl, phosphonyl, and other organic moieties conventionally used and accepted in organic chemistry.

As used herein, the term "alkyl" refers to saturated hydrocarbons having 1 to 12 carbon atoms, including straight chain, branched chain, 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, t-butyl, sec-butyl, isobutyl, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. The term alkyl includes unsubstituted alkyl groups and substituted alkyl groups. The term "C ₁-C_n alkyl" (where n is an integer from 2 to 12) represents an alkyl group having from 1 to the "n" carbon atoms shown. The alkyl residue may be substituted or unsubstituted. In some embodiments, for example, an alkyl group may be substituted with a hydroxyl, amino, carboxyl, carboxylate, amide, carbamate, or aminoalkyl group, or the like.

As used herein, "lower" in "lower aliphatic", "lower alkyl", "lower alkenyl" and "lower alkynyl" means that the moiety has at least one (at least two for alkenyl and alkynyl) and equal to or less than 6 carbon atoms unless the carbon number is limited.

The terms "cycloalkyl", "alicyclic", "carbocycle" and equivalents refer to a group comprising a saturated or partially unsaturated carbocycle in a monocyclic, spiro (sharing one atom) or fused (sharing at least one bond) carbocycle system, wherein the carbocycle system has 3 to 15 carbon atoms. 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, and the like. The term cycloalkyl includes unsubstituted cycloalkyl and substituted cycloalkyl. The term "C ₃-C_n cycloalkyl" wherein n is an integer from 4 to 15, means cycloalkyl having 3 to the "n" carbon atoms shown in the ring structure. As used herein, unless otherwise indicated, a "lower cycloalkyl" group refers to a group having at least 3 and equal to or less than 8 carbon atoms in its ring structure.

"Cycloalkyl residues" may be saturated or groups containing one or more double bonds in the ring system. In particular, they may be saturated or contain a double bond in the ring system. In unsaturated cycloalkyl residues, the double bond may be present at any suitable position. Monocycloalkyl residues include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl or cyclotetradecyl, which may also be substituted with C _1-4 alkyl. Examples of substituted cycloalkyl residues are 4-methylcyclohexyl and 2, 3-dimethylcyclopentyl. Examples of parent structures for the bicyclo system are norbornane, bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane and bicyclo [3.2.1] octane.

The term "heterocycloalkyl" and equivalents as used herein refers to a group comprising a saturated or partially unsaturated carbocycle having 3 to 15 carbon atoms in a monocyclic, spiro (sharing one atom) or fused (sharing at least one bond) carbocycle system, including groups of 1 to 6 heteroatoms (e.g., N, O, S, P) or heteroatom (e.g., NH, NRx (Rx is alkyl, acyl, aryl, heteroaryl or cycloalkyl), PO ₂、SO、SO₂, etc.). The heterocycloalkyl group may be attached to the C or to a heteroatom (e.g., through a nitrogen atom). Examples of heterocycloalkyl groups include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, tetrahydrodithioanyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thiazalkyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxapentanyl, thiazapyrrolidinyl, oxazepinyl, diazanyl, thiazanyl, 1,2,3, 6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxane, 1, 3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothiophenyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo [3,1,0] hexyl, 3-azabicyclo [4,1,0] heptyl, 3H-indolyl, quinolizinyl, and the like. The term heterocycloalkyl includes unsubstituted heterocycloalkyl and substituted heterocycloalkyl. The term "C ₃-C_n heterocycloalkyl", wherein n is an integer from 4 to 15, means a heterocycloalkyl having 3 to the "n" atoms indicated in the ring structure, including at least one hetero group or atom as defined above. As used herein, unless otherwise indicated, "lower heterocycloalkyl" means having at least 3 and equal to or less than 8 carbon atoms in its cyclic structure.

The terms "aryl" and "aryl ring" as used herein refer to aromatic groups having "4n+2" electrons (pi) in conjugated single or multiple ring systems (fused or non-fused), and having 6 to 14 ring atoms, wherein n is an integer from 1 to 3. The polycyclic ring system includes at least one aromatic ring. The aryl group may be attached directly or through a C ₁-C₃ alkyl (also known as arylalkyl or aralkyl). Examples of aryl groups include, but are not limited to, phenyl, benzyl, phenethyl, 1-phenylethyl, tolyl, naphthyl, biphenyl, terphenyl, indenyl, benzocyclooctenyl, benzocycloheptenyl, azulenyl, acenaphthylenyl, fluorenyl, phenanthryl, anthracenyl, and the like. The term aryl includes unsubstituted aryl and substituted aryl. The term "C ₆-C_n aryl" (where n is an integer from 6 to 15) denotes an aryl group having from 6 to the "n" carbon atoms shown in the ring structure, including at least one heterocyclic group or atom as defined above.

The terms "heteroaryl" and "heteroaryl ring" as used herein refer to aromatic groups having "4n+2" electrons (pi) in conjugated monocyclic or polycyclic ring systems (fused or unfused), where n is an integer from 1 to 3, and include one to six heteroatoms (e.g., N, O, S, P) or groups including heteroatoms (e.g., NH, NRx (Rx is alkyl, acyl, aryl, heteroaryl or cycloalkyl), PO ₂、SO、SO₂, etc.). The polycyclic ring system includes at least one heteroaromatic ring. Heteroaryl groups may be attached directly or through a C ₁-C₃ alkyl group (also known as heteroarylalkyl or heteroarylalkyl). Heteroaryl groups may be attached to a carbon or to a heteroatom (e.g., through a nitrogen atom). Examples of heteroaryl groups include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, tetrazolyl, furyl, thienyl; isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolidinyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, chromene, isochromene, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, pteridinyl, furanyl, benzofuranyl, benzothiazolyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinolinyl, quinolinonyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, carbazolyl, phenanthridinyl, acridinyl, perylenyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, dibenzofuranyl, and the like. The term heteroaryl includes unsubstituted heteroaryl and substituted heteroaryl. The term "C ₅-C_n heteroaryl", wherein n is an integer from 6 to 15, denotes heteroaryl groups having from 5 to the "n" atoms shown in the ring structure, including at least one heterocyclic group or atom as defined above.

The term "heterocycle" or "heterocyclic" as used herein includes heterocycloalkyl and heteroaryl. Examples of heterocycles include, but are not limited to, acridinyl, azepinyl, benzimidazolyl, benzofuranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzotriazole, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, 4αh-carbazolyl, carbolinyl, chromanyl, chromene, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro [2,3-b ] tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, 3H-indolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 5-oxadiazolyl; 1,3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazole, pyridinyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quininyl, tetrahydrofuranyl, tetrahydroisoquinolyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2, 5-thiadiazinyl, 1,2, 3-thiadiazolyl, 1,2, 4-thiadiazolyl, 1,2, 5-thiadiazolyl, 1,3, 4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thienyl, triazinyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, 1,2, 5-triazolyl, 3, 4-triazolyl, xanthenyl and the like. The term heterocycle includes unsubstituted heterocyclyl and substituted heterocyclyl.

The term "amine" or "amino" as used herein refers to an unsubstituted or substituted fragment of the general formula-NR _aR_b, wherein R _a and R _b are each independently hydrogen, alkyl, aryl, or heterocyclyl, or R _a and R _b together form a heterocycle with the nitrogen atom to which they are attached. The term amino refers to a compound or fragment in which at least one carbon or heteroatom is covalently bonded to a nitrogen atom. Thus, the terms "alkylamino" and "dialkylamino" as used herein refer to amine groups having one and at least two C ₁-C₆ alkyl groups, respectively, attached to the nitrogen atom. The terms "arylamino" and "diarylamino" include at least one or two aryl-bonded groups attached to a nitrogen atom. The term "amide" or "aminocarbonyl" refers to a compound or fragment in which the carbon of the carbonyl or thiocarbonyl group is attached to a nitrogen atom. The term "acylamino" refers to a structure in which an amino group is attached directly to an acyl group.

The term "alkylthio" refers to an alkyl group having a mercapto group attached thereto. Suitable alkylthio groups include groups having from 1 to about 12 carbon atoms, preferably from 1 to about 6 carbon atoms. The term "alkylcarboxy" as used herein refers to an alkyl group having a carboxy group attached thereto.

The term "alkoxy" or "lower alkoxy" as used herein refers to a structure in which an alkyl group is attached to an oxygen atom. Representative alkoxy groups include groups having from 1 to about 6 carbon atoms, such as methoxy, ethoxy, propoxy, t-butoxy, and the like. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, propoxy, butoxy, pentyloxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy, dichloromethoxy, trichloromethoxy, and the like. The term "alkoxy" includes unsubstituted or substituted alkoxy, and perhaloalkoxy and the like.

The term "carbonyl" or "carboxyl" as used herein means compounds and fragments containing a carbon attached to an oxygen atom through a double bond. Examples of carbonyl containing moieties include aldehydes, ketones, carboxylic acids, amides, esters, anhydrides, and the like.

The term "acyl" as used herein is a carbonyl structure in which the carbon atom of the carbonyl group is attached to hydrogen (i.e., formyl), an aliphatic group (C ₁-C₆ alkyl, C ₁-C₆ alkenyl, C ₁-C₆ alkynyl, e.g., acetyl), cycloalkyl (C ₃-C₈ cycloalkyl), heterocyclyl (C ₃-C₈ heterocycloalkyl and C ₅-C₆ heteroaryl), and aryl (C ₆ aryl, e.g., benzoyl). The acyl group may be an unsubstituted or substituted acyl group (e.g., salicyloyl group).

The term "solvate" refers to a physical association of a compound with one or more solvent molecules (whether organic or inorganic). The physical association includes hydrogen bonding. In some cases, the solvate can be isolated, for example, when one or more solvent molecules are incorporated into the crystal lattice. "solvate" includes solvent compounds in the solution phase and solvates that can be separated. Examples of "solvates" include, but are not limited to, hydrates, ethanolates, methanolates, hemiethanolates, and the like. The term "hydrate" refers to the physical association of a compound with a water molecule.

"Pharmaceutically acceptable salt" of a compound refers to a salt of a pharmaceutically acceptable compound. Salts of desirable compounds (basic, acidic or charged functional groups) may retain or improve the biological activity and properties of the parent compound as defined herein and are not biologically undesirable. Pharmaceutically acceptable salts may be those mentioned by Berge et al, "Pharmaceutical Salts", J.Pharm.Sci.66,1-19 (1977). Including but not limited to:

(1) Salts formed by adding acids to basic or positively charged functional groups, inorganic acids including hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, nitric acid, phosphoric acid, carbonates, and the like. Organic acids include acetic acid, propionic acid, lactic acid, oxalic acid, glycolic acid, pivalic acid, t-butyl acetic acid, β -hydroxybutyric acid, valeric acid, caproic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, succinic acid, malic acid, fumaric 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, cyclohexylsulfamic acid, benzenesulfonic acid, sulfanilic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 3-phenylpropionic acid, lauryl sulfonic acid, lauryl sulfuric acid, oleic acid, palmitic acid, stearic acid, lauric acid, pamoic acid, pantothenic acid, lactobionic acid, alginic acid, galacturonic acid, gluconic acid, glucoheptonic acid, glutamic acid, naphthoic acid, hydroxynaphthoic acid, salicylic acid, ascorbic acid, stearic acid, muconic acid, and the like.

(2) When acidic protons are present in the parent compound or are replaced by metal ions, a base may be added to give a salt. The metal ions include alkaline metal ions (e.g., lithium, sodium, potassium), alkaline earth metal ions (magnesium, calcium, barium) or other metal ions such as aluminum, zinc, iron, etc. Organic bases include, but are not limited to, N' -dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, piperazine, chloroprocaine, procaine, choline, lysine, and the like.

Pharmaceutically acceptable salts can be synthesized from the parent compound containing a basic or acidic fragment by conventional chemical methods. Typically, such salts are prepared by reacting a compound (free acid or base) with an isostoichiometric amount of base or acid in water or an organic solvent or in a mixture of both. Salts may be prepared in situ during the final isolation or purification of the pharmaceutical agent or by separately reacting the purified compound of the invention in free acid or base form with the corresponding base or acid desired and isolating the salt formed thereby. The term "pharmaceutically acceptable salts" also includes zwitterionic compounds comprising cationic groups covalently bonded to anionic groups, which are referred to as "inner salts". All acids, salts, bases and other ionic and nonionic forms are encompassed by the compounds of the present invention. For example, if the compound of the present invention is an acid, the salt form of the compound is also included. Also, if the compounds of the present invention are salts, the acid and/or base forms of the compounds are also included.

As used herein, the term "effective amount" refers to the amount or dose of a therapeutic agent (e.g., a compound) that provides a desired therapeutic, prophylactic, diagnostic, or prognostic effect in a subject after administration to the subject in a single dose or in multiple doses. The effective amount can be readily determined by the attending physician or diagnostician by known techniques and by observing results obtained under similar circumstances. In determining an effective amount or dose of a compound to be administered, a number of factors are considered, including, but not limited to: the weight, age, and general health of the subject; specific diseases involved; the degree of involvement or severity of the disease or condition to be treated; responses of the subject individual; the particular compound being administered; mode of administration; bioavailability characteristics of the administered formulation; a selected dosage regimen; use of concomitant medications; and other related considerations.

By "pharmaceutically acceptable" is meant that the term describes a drug, pharmaceutical product, inert ingredient, etc., suitable for use in contact with cells or tissues of humans and animals without undue toxicity, incompatibility, instability, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. Generally refers to compounds or compositions approved or approvable by a regulatory agency of the federal or a state government or listed in the U.S. pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

By "pharmaceutically acceptable carrier" is meant a diluent, adjuvant, excipient, carrier, or vehicle with which the compound is administered. The terms "pharmaceutically acceptable carrier" and "pharmaceutically acceptable carrier" are used interchangeably herein.

"Pharmaceutical composition" is meant to include a compound as described herein, and at least one component, including pharmaceutically acceptable carriers, diluents, adjuvants, excipients or vehicles, such as preserving, bulking, disintegrating, wetting, emulsifying, suspending, sweetening, flavoring, perfuming, antibacterial, antifungal, lubricating, dispersing agents and the like, depending on the mode of administration and the requirements of the dosage form. "preventing" or "prevention" is used to mean at least reducing the likelihood of acquiring a disease or disorder (or susceptibility) to acquire a disease or disorder (i.e., not allowing the clinical symptoms of at least one disease to develop into a patient that may be exposed to or susceptible to the disease but has not experienced or displayed symptoms of the disease).

In some embodiments, "treating" or "treating" any disease or disorder refers to alleviating at least one disease or disorder. In certain embodiments, "treating" or "treatment" refers to alleviating at least one physical parameter, which may be distinguishable or indistinguishable by the patient. In certain embodiments, "treatment" or "treatment" refers to inhibiting a disease or disorder physically (e.g., stabilization of a discernible symptom) or physiologically (e.g., stabilization of a physical parameter) or both. In certain embodiments, "treatment" or "treatment" refers to an adverse effect of improving quality of life or disease in a subject in need thereof. By "therapeutically effective amount" is meant an amount of a compound administered to a subject for treating or preventing a disease that is sufficient to achieve an effect of treating or preventing the disease. "therapeutically effective amount" will depend on the compound; disease and severity thereof; the age, weight, etc. of the subject to be treated or prevented from suffering from the disease. As used herein, a "therapeutically effective amount" refers to a compound or composition that is sufficient to prevent, treat, inhibit, reduce, alleviate or eliminate one or more etiologies, symptoms, or complications of a disease, such as cancer.

The term "prodrug" or its equivalent means an agent that is converted directly or indirectly to an active form in vitro or in vivo (see, e.g., R.B.Silverman,1992,"The Organic Chemistry of Drug Design and Drug Action,"Academic Press,Chap.8;Bundgaard,Hans;Editor.Neth.(1985),"Design of Prodrugs".360pp.Elsevier,Amsterdam;Stella,V.;Borchardt,R.;Hageman,M.;Oliyai,R.;Maag,H.;Tilley,J.(Eds.)(2007),"Prodrugs:Challenges and Rewards,XVIII,1470p.Springer). a prodrug may be used to alter the biological distribution (e.g., such that the agent does not normally enter the protease reaction site) or pharmacokinetics a variety of groups have been used to modify a compound to form a prodrug, such as an ester, ether, phosphate, etc., when the prodrug is administered to a subject, the group is cleaved off enzymatically or non-enzymatically, reduced, oxidized, or hydrolyzed, or otherwise liberates the active compound.

The term "ester" refers to a compound that may be represented by the formula RCOOR (carboxylate) or RSO ₃ R' (sulfonate), and that may generally be formed by the reaction between a carboxylic acid or sulfonic acid, respectively, and an alcohol (elimination of a molecule of water).

For the compounds provided herein, in some embodiments, salts, pharmaceutically acceptable salts thereof are also included. Those skilled in the art will be aware of the many possible salt forms (e.g., TFA salt, tetrazole salt, sodium salt, potassium salt, etc.), and may also select suitable salts based on considerations known in the art. The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases (including inorganic acids and bases and organic acids and bases). For example, for compounds containing basic nitrogen, salts thereof may be prepared with pharmaceutically acceptable non-toxic acids (including inorganic and organic acids). Pharmaceutically acceptable acids suitable for use in the present invention include, but are not limited to, acetic acid, benzenesulfonic acid (benzenesulfonate), benzoic acid, camphorsulfonic acid, citric acid, vinylsulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like. When the compound contains an acidic side chain, pharmaceutically acceptable bases suitable for use in the present invention include, but are not limited to, metal salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from lysine, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine.

Composition and method for producing the same

In one embodiment, a pharmaceutical composition is provided comprising a compound of the invention, e.g., a compound of formulas I-1 and a compound of table 1 or a pharmaceutically acceptable salt, ester, chelate, hydrate, or solvate thereof, and a pharmaceutically acceptable carrier. In yet another embodiment, a pharmaceutical composition comprising a compound of formula I and a compound of table 1, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier is provided.

Examples

The invention will be more readily understood by reference to the following examples, which are provided to illustrate the invention and should not be construed to limit the scope of the invention in any way.

Unless defined otherwise or the context clearly indicates otherwise, 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 belongs. It should be understood that any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention.

Example 1: preparation of N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridin) -2-thiazole) aminocarbonyl-L-proline- ¹⁸ O-amide (Compound 1 or Alpelisib- ¹⁸O₁)

Under nitrogen protection, (S) -1-N-Boc-2-pyrrolidinecarbonitrile (100 mg,0.51 mmol), palladium acetate (12 mg,0.051 mmol), 2-bipyridine (8 mg,0.051 mmol) were added to H ₂ ¹⁸ O (98% oxygen-18 abundance, 0.5 mL) and the reaction stirred in a lock at 60℃for 24 hours. The reaction solution was cooled to room temperature and transferred to a flask, concentrated in vacuo, and the residue was purified by column chromatography (mobile phase, dichloromethane: methanol=100/0 to 50/1) to finally give N-Boc-L-prol- ¹⁸ O-amide (48 mg; oxygen-18 abundance, 97.3%).

N-Boc-L-prolyl- ¹⁸ O-amide (48 mg,0.22 mmol) was dissolved in dichloromethane (0.5 mL), then a hydrochloric acid/dioxane solution (4M, 0.5 mL) was added, the reaction was stirred at 25℃for 1 hour, and then the reaction solution was dried under an oil pump to give L-prolyl- ¹⁸ O-amide hydrochloride (35 mg).

N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridine) -2-thiazole) aminocarbonyl- [1H ] -imidazole ester (79 mg,0.2 mmol) was added to DMF (1 mL), followed by L-prolyl- ¹⁸ O-amide hydrochloride (33 mg,0.22 mmol) and triethylamine (80 mg,0.8 mmol), and the reaction was stirred at 30℃for 16H. The reaction solution was dried by oil pump, and the residue was purified by column chromatography (mobile phase, dichloromethane: methanol=100/0 to 30/1) to give compound 1 (70 mg, oxygen-18 abundance 97.34%, yield 81.87%).

¹H NMR(500MHz,DMSO-d₆)：δppm 10.96(s,1H),8.60(d,J＝5.0Hz,1H),7.55(s,1H),7.41(s,2H),6.97(s,1H),4.26(s,0.48H),3.60(s,1H),3.46(s,1H),2.42(s,4H),2.09(s,1H),1.87(s,3H),1.61(s,6H).

¹³C NMR(125MHz,DMSO-d6)：δppm 174.36,159.87,158.99,152.99,149.58,145.69,141.44,129.94,127.69,121.80,120.78,60.36,55.40,46.92,46.71,30.43,21.93,16.89.

m/z(ESI⁺)：443.7;m/z(ESI^-)441.5。

Example 2: preparation of N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridin) -2-thiazole) aminocarbonyl-L-proline- ¹⁷ O-amide (Compound 2 or Alpelisib- ¹⁷O₁)

Compound 2 was obtained in the same manner as in example 1, except that O ¹⁷ -water (H ₂ ¹⁷ O) was used instead of H ₂ ¹⁸ O used in example 1.

Example 3: preparation of N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridin) -2-thiazole) aminocarbonyl-L-proline- ¹³C₁ -amide (Compound 4 or Alpelisib- ¹³C₁)

Compound 4 was obtained in a similar manner to example 1, except that proline- ¹³C₁ -amide hydrochloride was used as starting material.

Example 4: preparation of N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridin) -2-thiazole) aminocarbonyl-L-prolinamide- ¹⁵ N (Compound 5 or Alpelisib- ¹⁵ N)

L-proline (5 g,43.4mmol,1.0 eq.) and triethylamine (6.6 g,65.2mmol,1.5 eq.) were dissolved in 50ml methanol and (Boc) ₂ O (9.5 g,43.5mmol,1 eq.) was slowly added and the mixture stirred at 50℃for 2h. The reaction was concentrated, and the residue was purified by column chromatography (eluent: meOH/dcm=1/50) to give N-t-butoxycarbonyl-L-proline (6 g, 64.2%).

N-t-Butoxycarbonyl-L-proline (1.5 g,6.97mmol,1.0 eq.) obtained in the previous step was dissolved in 1, 4-dioxane (27 mL), pyridine (0.36 mL,4.20mmol,0.6 eq.) was added, ¹⁵ N-ammonium sulfate (1.19 g,8.87mmol,1.3 eq.) and (Boc) ₂ O (1.98 g,8.87mmol,1.3 eq.) were added and the reaction mixture was stirred overnight at 25 ℃. The reaction solution was concentrated, and the residue was purified by column chromatography (eluent: meOH/dcm=100/0 to 50/1) to give N-t-butoxycarbonyl-L-prol- ¹⁵ N-amide (0.29 g; ¹⁵ N abundance, 99.4%).

The thus obtained N-t-butoxycarbonyl-L-prol- ¹⁵ N-amide (0.29 g, 1.349 mmol,1.0 eq.) was dissolved in DCM (3 mL), 4M HCl/dioxane solution (1.4 mL,5.6mmol,4.16 eq.) was added and stirred at room temperature for 0.5h. A white solid precipitated, was filtered and dried under vacuum to give L-prolyl- ¹⁵ N-amide hydrochloride (0.16 g; ¹⁵ N abundance, 99.4%).

4-Methyl-5- (2- (1, 1-trifluoro-2-methylpropan-2-yl) pyridin-4-yl) thiazol-2-amine (0.5 g,1.66mmol,1.0 eq.) was dissolved in DCM (25 mL) and CDI (0.4 g,2.46mmol,1.5 eq.) was added and stirred at 40℃for 4h. A large amount of white solid precipitated, was filtered and dried under vacuum to give N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridin) -2-thiazole) aminocarbonyl- [1H ] -imidazolate (0.5 g, 76.9% yield).

L-prolyl- ¹⁵ N-amide hydrochloride (0.16 g,1.06mmol,1.05 eq.) and N- (4-methyl-5- (2, 2-trifluoro-1, 1-dimethylethyl) -4-pyridine) -2-thiazole) aminocarbonyl- [1H ] -imidazole ester (400 mg,1.01mmol,1.0 eq.) were dissolved in pyridine (5 mL) and DMAP (3 mg,0.016mmol,0.023 eq.) was added and stirred overnight at 25 ℃. The reaction mixture was concentrated, and the residue was purified by column chromatography (eluent: DCM/meoh=100/0 to 30/1) to give compound 5 (240 mg, ¹⁵ N-abundance 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)：δppm 16.78,21.91,24.39,30.41,46.72,46.92,60.38,120.68,121.71,125.39,127.65,129.90,132.15,141.42,149.53,159.00,174.21,174.32.

m/z(ESI^-)440.6;m/z(ESI⁺)442.8。

Example 5: pharmacokinetic comparative study of Compounds 1 and Alpelisib in SD rats

A group of 6 SD rats was used for the experiment. Compound 1 (Alpelisib- ¹⁸O₁) was mixed with Alpelisib in a molar ratio of 1:1 to give a dosing solution, wherein the concentration of both in the dosing solution was 1.25mg/mL. The administration solution was administered to animals by gavage at a dose of 2.5mg/kg. Blood samples were taken at a series of preset time points after administration (time points 0.167, 0.5, 1,2, 3,4, 6, 8 and 24 hours after administration, respectively). The blood samples were converted to plasma samples using the general procedure and the concentrations of compounds 1 and Alpelisib in the samples were analyzed by LC-MS/MS. The data obtained are shown in Table 2 and FIG. 1. In FIG. 1, the symbols- -O- -and- -A- -represent the drug concentration-time curves in plasma for Compounds 1 and Alpelisib, respectively, after an equal dose of oral administration.

Table 2. PK parameters of compounds 1 and Alpelisib in SD rats.

Parameters (parameters)	Unit (B)	Compound 1	Alpelisib
				AUC_0-t	ug/L*h	28719	26405
AUC_0-∞	ug/L*h	28798	26476
				Tmax	H	3	3
Vz/F	L/kg	1.655	1.798
				CLz/F	L/h/kg	0.434	0.472
Cmax	ug/L	2390	2297

Example 6: pharmacokinetic comparative study of Compounds 1 and Alpelisib in ICR mice

48 ICR mice were randomly divided into 6 groups (8 per group). Compound 1 (Alpelisib- ¹⁸O₁) was combined with Alpelisib in a 1:1 molar ratio to give a dosing solution, wherein the concentration of both in the dosing solution was 1.25mg/mL. The administration solution was administered to animals by gavage at a dose of 2.5mg/kg. After administration, blood samples were taken at a series of preset time points (time points 0.5, 1,2, 4, 8 and 12 hours after administration, respectively), each time point being sampled from a group of animals. The blood samples were converted to plasma samples using the general procedure and the concentrations of compounds 1 and Alpelisib in the samples were analyzed by LC-MS/MS. The data obtained are shown in Table 3 and FIG. 2. In FIG. 2, the symbols- -O- -and- -A- -represent the drug concentration-time curves in plasma for Compounds 1 and Alpelisib, respectively, after an equal dose of oral administration.

Table 3 PK parameters in ICR mice for Compounds 1 and Alpelisib.

Parameters (parameters)	Unit (B)	Compound 1	Alpelisib
				AUC(0-t)	ug/L*h	27845	26212
AUC(0-∞)	ug/L*h	32554	30597
				Tmax	h	2	2
Vz/F	L/kg	2.236	2.37
				CLz/F	L/h/kg	0.384	0.409
Cmax	ug/L	4519	4208

Example 7: production of metabolites M4 and M4- ¹⁸O₁ in liver S9 medium.

A stock solution (50. Mu.L) of the mixture containing equimolar amounts of compounds 1 and Alpelisib was diluted with 0.1M Tris-acetate buffer pre-heated in a 37℃water bath to a working solution with a compound concentration of 4.5. Mu.M. To each incubation well, the above working solution (50. Mu.L) and liver S9 solution (50. Mu.L) were added and mixed well. After 5 minutes of pre-incubation, β -NADPH solution (100. Mu.L) was added to each incubation well. The incubation plates were then incubated at 37 ℃ and samples were collected and analyzed at time points of 0, 15, 30, 60, and 240 minutes.

Sample analysis method: three samples were collected at each time point, mixed well (Vertex) after adding stop solution (800. Mu.L) to the samples at each preset time point, centrifuged at 12000rpm for 5 minutes, and the supernatant was transferred to an analysis sample tube and analyzed for M4 and M4- ¹⁸O₁ concentrations in the samples by LC-MS/MS.

Under the same experimental conditions, the following two experiments were performed in parallel: (1) A series of dilutions of the mixture solutions of M4 and M4- ¹⁸O₁ at equimolar concentrations were performed and M4- ¹⁸O₁ in each solution were analyzed by LC-MS/MS, demonstrating that M4 and M4- ¹⁸O₁ have the same response values and are linear under this condition; (2) When only compound 1 was incubated, no M4 was produced, only M4- ¹⁸O₁ was produced.

The livers S9 used in the experiment were (a) monkey livers S9, (b) mixed human livers S9, (c) mini pig livers S9, and (d) male Wistar rat livers 9, respectively. Fig. 3 (fig. 3a to 3 d) is a summary of experimental results.

Although the present invention has been described in detail with reference to the embodiments thereof, these embodiments are provided for the purpose of illustration and not limitation of the invention. Other embodiments that can be obtained according to the principles of the present invention fall within the scope of the invention as defined in the claims.

The contents of all documents and documents listed herein are incorporated by reference in their entirety.

Claims

1. Use of a compound of formula (I) or a pharmaceutically acceptable salt, ester, hydrate, chelate or solvate thereof, in a medicament for the prevention or treatment of PIK3 CA-related PROS disease:

r ¹ and R ² are independently hydrogen or a protecting group.

2. Use of a compound according to claim 1, wherein the compound is selected from the group of compounds shown below:

3. The use of a compound of claim 1, wherein the level of heavy isotopic enrichment in the compound is about 5% or more, about 10% or more, about 20% or more, about 50% or more, about 75% or more, about 90% or more, about 95% or more, about 97% or more, about 98% or more, or about 99% or more.

4. Use of a pharmaceutical composition comprising a compound as defined in any one of claims 1-3 and a pharmaceutically acceptable carrier in the manufacture of a medicament for the treatment or prevention of a PROS disorder.

5. The compound according to any one of claims 1 to 3 and the use of the composition according to claim 4, wherein the PROS disease is congenital lipid overgrowth, vascular malformations, epidermal nevi and spinal/skeletal abnormalities and/or scoliosis (cage) syndrome.

6. A compound according to any one of claims 1 to 3 and the use of a composition according to claim 4, wherein the PROS disease is fibrous fat overgrowth (FAO), hyperproliferative adiposity (HHML), macrofmger deformity, fibrous fat invasive lipoma, megabrain-capillary deformity (MCAP) or Dysplastic Megabrain (DMEG).

7. The use of a compound according to any one of claims 1 to 3 and a composition according to claim 4, wherein the PROS disorder is Klippel-Trenaunay syndrome.

8. The use of a compound according to any one of claims 1 to 3 and a composition according to claim 4, wherein the subject of the compound is a mammal.

9. The use of claim 8, wherein the mammal is a human.

10. A kit comprising a compound according to any one of claims 1 to 3 or a pharmaceutical composition according to claim 4, and instructions for its use.