HK1218757B - Pyrazole compounds as modulators of fshr and uses thereof - Google Patents

Description

Pyrazole compounds used as follicle-stimulating hormone receptor modulators and their uses

Related applications

This patent application claims priority to U.S. Provisional Patent Application No. 61/838,460, filed on June 24, 2013, and U.S. Provisional Patent Application No. 61/898,608, filed on November 1, 2013, which are hereby incorporated by reference in their entireties.

Technical Field

The present invention relates to pyrazole compounds that are useful as follicle stimulating hormone receptor (FSHR) agonists. The present invention also provides pharmaceutically acceptable compositions comprising the compounds of the present invention and methods of using the compositions to treat various diseases.

Background of the Invention

Gonadotropins play an important role in many bodily functions, including metabolism, temperature regulation, and reproduction. Gonadotropins act on specific types of gonadal cells to initiate ovarian and testicular differentiation and steroid production. The gonadotropin FSH (follicle-stimulating hormone) is released from the anterior pituitary gland under the influence of gonadotropin-releasing hormone and testosterone, and from the placenta during pregnancy. FSH is a heterodimeric glycoprotein hormone with a similar structure to luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and chorionic gonadotropin (CG). LH and TSH are produced in the pituitary gland, while CG is produced in the placenta. In women, FSH plays a central role in stimulating follicular development and maturation. Furthermore, it is the primary hormone regulating testosterone secretion, with LH inducing ovulation. In men, FSH is responsible for the integrity of the seminiferous tubules and acts on Sertoli cells to support gametogenesis.

The hormones are relatively large (28-38 kDa) and consist of a common α-subunit non-covalently linked to different β-subunits, which confer receptor binding specificity. The cellular receptors for these hormones are expressed on the Sertoli cells of the testis and the granulosa cells of the ovary. The FSH receptor is known to be a member of a class of membrane-bound receptors that bind to G proteins and, when activated, stimulates increased activity of adenylyl cyclase. This leads to increased levels of the intracellular second messenger adenosine 3',5'-monophosphate (cAMP), which in turn leads to increased steroid synthesis and secretion. Hydropathicity analysis of the amino acid sequences of these receptors reveals three general domains: a hydrophilic amino-terminal region, which is believed to constitute the amino-terminal extracellular domain; seven hydrophobic segments spanning the membrane length, which are believed to constitute the transmembrane domain; and a carboxy-terminal region containing potential phosphorylation sites (serine, threonine, and tyrosine residues), which is believed to constitute the carboxy-terminal intracellular or cytoplasmic domain. The glycoprotein hormone receptor family differs from other G protein-bound receptors (eg, beta-2-adrenergic receptor, rhodopsin receptor, and substance K receptor) by the larger size of the hydrophilic amino-terminal domain involved in hormone binding.

Each year, 2.4 million couples in the United States experience infertility and require treatment. FSH, extracted from urine or a protein product produced using recombinant DNA technology, is administered parenterally to induce ovulation and control ovarian hyperstimulation. Ovulation induction involves ovulation of a single follicle, while control ovarian hyperstimulation involves obtaining multiple oocytes for use in various in vitro assisted reproductive technologies, such as in vitro fertilization (IVF). FSH is also used clinically to treat male hypogonadism and male infertility, such as certain conditions that prevent sperm production.

FSHR is a highly specific target during follicular growth and is expressed only in the ovary. However, the use of FSH is limited due to its high cost, lack of oral dosage forms, and the need for close monitoring by a specialized physician. Therefore, there is a desire to develop and identify a non-peptide small molecule that can replace FSH and can be taken orally. Low-molecular-weight FSH mimetics with agonist properties have been disclosed in International Patent Applications WO 2002/09706 and WO 2010/136438, as well as in U.S. Patent No. 6,653,338. However, there is still a need for low-molecular-weight hormone mimetics that can selectively activate FSHR.

Summary of the Invention

It has been found that the compounds of the present invention and pharmaceutically acceptable compositions thereof are effective FSHR modulators. The compounds are represented by the general formula I:

or a pharmaceutically acceptable salt thereof, wherein Ring A, X, Y, Z, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , n and p have the meanings defined and described in the Examples herein.

The compounds of the present invention and pharmaceutically acceptable compositions thereof can be used to treat various diseases, disorders or conditions associated with abnormal cellular responses triggered by follicle stimulating hormone events, including those described herein.

Detailed Description of Certain Embodiments

1. General Definition of the Compounds of the Invention

In certain embodiments, the present invention provides modulators of the follicle stimulating hormone receptor (FSHR). In certain embodiments, the present invention provides positive allosteric modulators of FSHR. In some embodiments, such compounds include those represented by the general formula described herein, or pharmaceutically acceptable salts thereof, wherein the variables are defined and described.

2. Compounds and Definitions

The compounds of the present invention include those summarized above and further described by the classes, subclasses, and species disclosed herein. Unless otherwise indicated, the following definitions as used herein shall apply. For purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics, 75th edition). In addition, general principles of organic chemistry are described in "Organic Chemistry" (Thomas Sorrell, University Science Books, Sausalito: 1999) and "March's Advanced Organic Chemistry" (5th edition, eds. Smith, M.B. and March, J., John Wiley & Sons, New York: 2001), the entire contents of which are incorporated herein by reference.

As used herein, the term "aliphatic" or "aliphatic group" refers to a substituted or unsubstituted straight (i.e., unbranched) or branched hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic, with a single point of attachment to the rest of the molecule. Unless otherwise specified, an aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-4 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-3 aliphatic carbon atoms, and in some embodiments, an aliphatic group contains 1-2 aliphatic carbon atoms. In some embodiments, a "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic _C3 - _C6 hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic, with a single point of attachment to the rest of the molecule. Exemplary aliphatic groups are linear or branched, substituted or unsubstituted C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

The term "lower alkyl" refers to a C _1-4 straight or branched chain alkyl group. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl and tert-butyl.

The term "lower haloalkyl" refers to a C _1-4 straight or branched chain alkyl group containing one or more halogen atoms.

The term "heteroatom" refers to one or more oxygen, sulfur, nitrogen, or phosphorus (including any oxidized form of nitrogen, sulfur, or phosphorus; the quaternized form of any basic nitrogen; a substitutable nitrogen of a heterocycle, such as N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR ⁺ (as in N-substituted pyrrolidinyl)).

As used herein, the term "unsaturated" refers to a moiety having one or more units of unsaturation.

As used herein, the term "divalent C _1-8 (or C _1-6 ) saturated or unsaturated linear or branched hydrocarbon chain" refers to divalent alkylene, alkenylene, alkynylene chains, which are linear or branched as defined herein.

The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -( _CH2 ) _n- , where n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by a substituent. Suitable substituents include those described below for substituted aliphatic groups.

The term "alkenylene" refers to a divalent alkenyl group. Substituted alkenylene chains are polymethylene groups containing at least one double bond in which one or more hydrogen atoms are replaced by a substituent. Suitable substituents include those described below for substituted aliphatic groups.

The term "halogen" refers to F, Cl, Br or I.

The term "aryl" used alone or as part of a larger moiety such as "aralkyl", "arylhydroxy" or "aryloxyalkyl" refers to monocyclic and bicyclic ring systems having a total of 5 to 14 ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" is used interchangeably with the term "aryl ring". In certain embodiments of the present invention, "aryl" refers to an aromatic ring system. Exemplary aryl groups are phenyl, biphenyl, naphthyl, anthracenyl, and the like, which optionally include one or more substituents. As used herein, the term "aryl" also includes within its scope groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl.

The terms "heteroaryl" and "heteroaryl-", used alone or as part of a larger moiety such as "heteroaralkyl" or "heteroarhydroxy", refer to radicals having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; 6, 10, or 14 pi electrons shared in the cyclic array; and 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur and any quaternized form of a basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroar-" also include groups formed by the fusion of a heteroaromatic ring with one or more aromatic, cycloaliphatic, or heterocyclic rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic or bicyclic. The term "heteroaryl" is used interchangeably with the term "heteroaryl ring" or "heteroaromatic group," any of which includes rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl, wherein the alkyl and heteroaryl portions are independently optionally substituted.

As used herein, the terms "heterocycle,""heterocyclyl,""heterocyclicgroup," and "heterocyclic ring" are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more, preferably 1 to 4, heteroatoms as defined above in addition to carbon atoms. The term "nitrogen," when used with respect to a ring atom of a heterocycle, includes substituted nitrogen. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or ⁺ NR (as in N-substituted pyrrolidinyl).

A heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that forms a stable structure, and any ring atom may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclic group," and "heterocyclic moiety" are used interchangeably herein and also include groups in which a heterocyclic ring is fused to one or more aromatic, heteroaromatic, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclic ring. A heterocyclic group may be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

As described herein, certain compounds of the present invention may contain "optionally substituted" moieties. In general, the term "substituted," whether or not preceded by the term "optionally," means that one or more hydrogen atoms of the designated moiety are replaced with a suitable substituent. "Substituted" applies to one or more hydrogen atoms, either explicitly or implicitly, of the structure (e.g., at least and at least unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from the specified group, the substituents at each position may be the same or different. Combinations of substituents envisioned by the present invention are preferably those that result in stable or chemically feasible compounds. As used herein, the term "stable" refers to compounds that do not undergo substantial alteration when subjected to conditions that allow their manufacture, detection, and, in certain embodiments, recovery, purification, and use for one or more of the purposes disclosed herein.

Suitable monovalent substituents on the substitutable carbon atoms of the “optionally substituted” group are independently deuterium; halogen; –(CH ₂ ) _0-4 R°; –(CH 2 ) _0-4 OR°; –O(CH ₂ ) _0-4 R°, –O–(CH _{2 ) 0-4} C(O)OR°; –(CH ₂ ) _0-4 CH(OR°) ₂ ; –(CH ₂ ) _{0-4 SR} °; –(CH ₂ ) _0-4 Ph, which may be substituted by R°; –(CH ₂ ) _0-4 O(CH ₂ ) _0-1 Ph, which may be substituted by R°; –CH═CHPh, which may be substituted by R°; –(CH ₂ ) _0-4 O(CH ₂ ) _0-1 -pyridyl, which may be substituted by R°; –NO ₂ ; –CN; –N ₃ ; –(CH ₂ ) _0–4 N(R°) ₂ ;–(CH ₂ ) _0–4 N(R°)C(O)R°;–N(R°)C(S)R°;–(CH ₂ ) _0–4 N(R°)C(O)NR° ₂ ;-N(R°)C(S)NR° ₂ ;–(CH ₂ ) _0–4 N(R°)C(O)OR°; –N(R°)N(R°)C(O)R°; –N(R°)N(R°)C(O)NR° ₂ ; –N(R°)N(R°)C(O)OR°; –(CH ₂ ) _0–4 C(O)R°; –C(S)R°; –(CH ₂ ) _0–4 C(O)OR°; –(CH ₂ ) _0–4 C(O)SR°;-(CH ₂ ) _0–4 C(O)OSiR° ₃ ; –(CH ₂ ) _0–4 OC(O)R°; –OC(O)(CH ₂ ) _0–4 SR°, SC(S)SR°; –(CH ₂ ) _0–4 SC(O)R°; –(CH ₂ ) _0–4 C(O)NR° ₂ ; –C(S)NR° ₂ ;–C(S)SR°;–SC(S)SR°,-(CH ₂ ) _0–4 OC(O)NR° ₂ ;-C(O)N(OR°)R°;–C(O)C(O)R°;–C(O)CH ₂ C(O)R°;–C(NOR°)R°;-(CH ₂ ) _0–4 SSR°;–(CH ₂ ) _0–4 S(O) ₂ R°;–(CH ₂ ) _0–4 S(O) ₂ OR°；–(CH ₂ ) _0–4 OS(O) ₂ R°；–S(O) ₂ NR° ₂ ；–(CH ₂ ) _0–4 S(O)R°；–N(R°)S(O) ₂ NR° ₂ ；–N(R°)S(O) ₂ R°；–N(OR°)R°；–C(NH)NR° ₂ ；–P(O) ₂ R°；–P(O)R° ₂ ；–OP(O)R° ₂ ；–OP(O)(OR°) ₂ ；SiR° ₃ ；–(C _1–4 linear or branched alkylene)O–N(R°) ₂ ；or (C _1–4 linear or branched alkylene)C(O)O–N(R°) ₂ ， wherein each R° may be substituted as defined below and is independently hydrogen, C _1-6 aliphatic, –CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, -CH ₂ -(5-6 membered heteroaryl ring), or a 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, regardless of the above definition, two independently occurring R° together with the atoms intervening therebetween form a 3-12 membered saturated, partially unsaturated or aromatic monocyclic or polycyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R° (or the ring formed by two independently occurring R° together with the intervening atoms) are independently deuterium, halogen, –(CH ₂ ) _0–2 R ^● , –(haloR ^● ), –(CH ₂ ) _0–2 OH, –(CH ₂ ) _0–2 OR ^● , –(CH ₂ ) _0–2 CH(OR ^● ) ₂ , –O(haloR ^● ), –CN, –N ₃ , –(CH ₂ ) _0–2 C(O)R ^● , –(CH ₂ ) _0–2 C(O)OH, –(CH ₂ ) _0–2 C(O)OR ^● , –(CH ₂ ) _0–2 SR ^● , –(CH ₂ ) _0–2 SH, –(CH ₂ ) _0–2 NH ₂ , –(CH ₂ ) _0–2 NHR ^● , –(CH ₂ ) _0-2 NR ^● ₂ , –NO ₂ , –SiR ^● ₃ , –OSiR ^● ₃ , –C(O)SR ^● , –(C _1-4 straight or branched alkylene)C(O)OR ^● , or –SSR ^● , wherein each R ^● is unsubstituted or, when preceded by “halo”, substituted only with one or more halogens, and is independently selected from C _1-4 aliphatic, –CH ₂ Ph, –O(CH ₂ ) _0-1 Ph, or a 5-6 membered saturated, partially unsaturated, or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an “optionally substituted” group include the following: ═O, ═S, ═NNR ^* ₂ , ═NNHC(O)R ^* , ═NNHC(O)OR ^* , ═NNHS(O) ₂ R ^* , ═NR ^* , ═NOR ^* , —O(C(R ^* ₂ )) _2-3 O—, or —S(C(R ^* ₂ )) _2-3 S—, wherein each independent occurrence of R ^* is selected from hydrogen, a C _1-6 aliphatic group which may be substituted as defined below, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents bound to an ortho-substitutable carbon of an "optionally substituted" group include: -O(CR ^* ₂ ) _2-3 O-, wherein each independently occurring R ^* is selected from hydrogen, a C _1-6 aliphatic group which may be substituted as defined below, or an unsubstituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

Suitable substituents on the aliphatic group of R ^* include halogen, –R ^● , –(haloR ^● ), –OH, –OR ^● , –O(haloR ^● ), –CN, –C(O)OH, –C(O)OR ^● , –NH ₂ , –NHR ^● , –NR ^● ₂ , or –NO ₂ , wherein each R ^● is unsubstituted or, when preceded by “halo”, substituted only with one or more halogens, and is independently a C _1-4 aliphatic group, –CH ₂ Ph, –O(CH ₂ ) _{0– 1} Ph, or a 5-6 membered saturated, partially unsaturated, or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

Suitable substituents on the substitutable nitrogen of an "optionally substituted" group include those wherein each is independently hydrogen, a _C1-6 aliphatic group which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, regardless of the above definitions, two independently occurring substituents taken together with their intervening atoms form an unsubstituted 3-12 membered saturated, partially unsaturated, or aromatic monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group are independently halogen, –R ^● , –(haloR ^● ), –OH, –OR ^● , –O(haloR ^● ), –CN, –C(O)OH, –C(O)OR ^● , –NH ₂ , –NHR ^● , –NR ^● ₂ , or –NO ₂ , wherein each R ^● is unsubstituted or, when preceded by “halo”, is substituted only with one or more halogens, and is independently a C _1-4 aliphatic group, –CH ₂ Ph, –O(CH ₂ ) _{0– 1} Ph, or a 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

In certain embodiments, the terms "optionally substituted," "optionally substituted alkyl," "optionally substituted alkenyl," "optionally substituted alkynyl," "optionally substituted carbocycle," "optionally substituted aryl," "optionally substituted heteroaryl," "optionally substituted heterocycle," and any other optionally substituted groups used herein refer to unsubstituted or substituted groups wherein one, two, three or more hydrogen atoms are independently replaced by typical substituents, including but not limited to:

-F, -Cl, -Br, -I, deuterium,

-OH, protected hydroxy, alkoxy, oxo, thiooxo,

-NO ₂ , -CN, CF ₃ , N ₃ ,

-NH ₂ , protected amino, -NH alkyl, -NH alkenyl, -NH alkynyl, -NH cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocyclyl, -dialkylamino, -diarylamino, -diheteroarylamino,

-O-alkyl, -O-alkenyl, -O-alkynyl, -O-cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocyclyl,

-C(O)-alkyl, -C(O)-alkenyl, -C(O)-alkynyl, -C(O)-carbocyclyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocyclyl,

-CONH ₂ , -CONH-alkyl, -CONH-alkenyl, -CONH-alkynyl, -CONH-carbocyclyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocyclyl,

-OCO ₂ -alkyl, -OCO ₂ -alkenyl, -OCO ₂ -alkynyl, -OCO 2 -carbocyclyl, -OCO ₂ -aryl, -OCO ₂ -heteroaryl, -OCO ₂ -heterocyclyl, -OCONH ₂ , -OCONH-alkyl, -OCONH-alkenyl, -OCONH- _alkynyl , -OCONH-carbocyclyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocyclyl,

-NHC(O)-alkyl, -NHC(O)-alkenyl, -NHC(O)-alkynyl, -NHC(O)-carbocyclyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocyclyl, -NHCO ₂ -alkyl, -NHCO ₂ -alkenyl, -NHCO ₂ -alkynyl, -NHCO ₂ -carbocyclyl, -NHCO 2 -aryl, -NHCO ₂ -heteroaryl, -NHCO ₂ -heterocyclyl, -NHC(O)NH ₂ , -NHC(O)NH- _alkyl , -NHC(O)NH-alkenyl, -NHC(O)NH-alkenyl, -NHC(O)NH-carbocyclyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocyclyl, NHC(S)NH ₂ , -NHC(S)NH-alkyl, -NHC(S)NH-alkenyl, -NHC(S)NH-alkynyl, -NHC(S)NH-carbocyclyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocyclyl, -NHC(NH)NH ₂ , -NHC(NH)NH-alkyl, -NHC(NH)NH-alkenyl, -NHC(NH)NH-alkenyl, -NHC(NH)NH-carbocyclyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocyclyl, -NHC(NH)-alkyl, -NHC(NH)NH-alkenyl, -NHC(NH)NH-alkenyl, -NHC(NH)NH-carbocyclyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocyclyl, -NHC(NH)-alkyl, -NHC(NH)-alkenyl, -NHC(NH)-alkenyl, -NHC(NH)-carbocyclyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)NH-heterocyclyl,

-C(NH)NH-alkyl, -C(NH)NH-alkenyl, -C(NH)NH-alkynyl, -C(NH)NH-carbocyclyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocyclyl,

-S(O)-alkyl, -S(O)-alkenyl, -S(O)-alkynyl, -S(O)-carbocyclyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocyclyl-SO ₂ NH ₂ , -SO ₂ NH-alkyl, -SO ₂ NH-alkenyl, -SO 2 NH-alkynyl, -SO ₂ NH-carbocyclyl, -SO ₂ NH-aryl, -SO ₂ NH-heteroaryl, -SO ₂ NH-heterocyclyl _,

-NHSO ₂ -alkyl, -NHSO ₂ -alkenyl, -NHSO ₂ -alkynyl, -NHSO ₂ -carbocyclyl, -NHSO ₂ -aryl, -NHSO ₂ -heteroaryl, -NHSO ₂ -heterocyclyl,

-CH ₂ NH ₂ , -CH ₂ SO ₂ CH ₃ ,

-mono-, di-, or tri-alkylsilyl groups,

-alkyl, -alkenyl, -alkynyl, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -cycloalkyl, -carbocyclyl, -heterocyclyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, -S-alkyl, -S-alkenyl, -S-alkynyl, -S-carbocyclyl, -S-aryl, -S-heteroaryl, -S-heterocyclyl, or methylthiomethyl.

As used herein, the term "pharmaceutically acceptable salt" is intended to refer to salts that are suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reaction, or other problems or complications, and that are commensurate with a reasonable benefit/risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, the contents of which are incorporated by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic acids and bases, as well as organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed of an amino group with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or salts formed by other methods in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, gluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hydroiodide, 2-hydroxyethanesulfonate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.

Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Other pharmaceutically acceptable salts include suitable non-toxic ammonium salts, quaternary ammonium salts and amine cations formed using, for example, halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates.

Unless otherwise indicated, structures depicted herein are also meant to include all isomeric forms of the structure (e.g., enantiomers, diastereomers, and geometric (or conformational) isomers); for example, R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, individual stereochemical isomers as well as mixtures of enantiomers, diastereomers, and geometric (or conformational) isomers of the compounds of the present invention are within the scope of the present invention. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

In addition, unless otherwise indicated, structures described herein include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures include replacement of hydrogen by deuterium or tritium, or replacement of carbon by a ¹³ C- or ¹⁴ C-enriched carbon, and such compounds are within the scope of the present invention. In some embodiments, a group includes one or more deuterium atoms.

Compounds of Formula I are also intended to include isotopically labeled forms thereof. Isotopically labeled forms of compounds of Formula I differ from the compounds described herein only in that one or more atoms of the compound are replaced by one or more atoms having an atomic mass or mass number different from that of the atoms typically found in nature. Examples of isotopes that are readily commercially available and can be incorporated into compounds of Formula I by known methods include hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as ^2H , ^3H , ^13C , ^14C , ^15N , ^18O , 17O, 31P, ^32P , ^35S , ^18F , and ^36CI , ^respectively . Compounds of Formula I, prodrugs thereof, or pharmaceutically acceptable salts thereof containing one or more of ^the aforementioned isotopes and/or isotopes of other atoms are to be understood as part of the present invention. Isotopically labeled compounds of Formula I can be used in a variety of advantageous ways. For example, isotope-labeled compounds of Formula 1 incorporating radioisotopes such as ³ H or ¹⁴ C can be used in drug and/or substrate tissue distribution assays. These two radioisotopes, tritium ( ³ H) and carbon-14 ( ¹⁴ C), are particularly preferred due to their ease of preparation and good detectability. Since heavier isotopes such as deuterium ( ² H) have higher metabolic stability, incorporating such isotope-labeled compounds into compounds of Formula I is therapeutically beneficial. Higher metabolic stability directly leads to an increase in in vivo half-life or a reduction in dosage, which in most cases represents a preferred embodiment of the present invention. Isotope-labeled compounds of Formula I can generally be prepared by carrying out the steps disclosed in the synthetic schemes and related descriptions in the Examples and Preparations sections of this text, substituting readily available isotope-labeled reactants for non-isotope-labeled reactants.

In order to control the oxidative metabolism of a compound through the primary kinetic isotope effect, deuterium ( ^2H ) can be incorporated into the compound. The primary kinetic isotope effect is a change in the rate of a chemical reaction due to the replacement of an isotopic nucleus, which is caused by a change in the ground state energy required to form a covalent bond after the isotope replacement. Replacement of a heavier isotope generally results in a decrease in the ground state energy of a chemical bond, thereby causing a decrease in the rate of the rate-limiting bond cleavage reaction. If bond cleavage occurs in or near a saddle point region along the coordinates of a multi-product reaction, the product distribution ratio can be significantly altered. This is explained as follows: If deuterium is bonded to a non-replaceable position on a carbon atom, the rate difference _km / _kd is generally 2-7. If this rate difference is successfully applied to a compound of formula I that is easily oxidized, the in vivo properties of the compound can be significantly altered, thereby improving pharmacokinetic properties.

When discovering and developing therapeutic agents, those skilled in the art attempt to optimize pharmacokinetic parameters while maintaining favorable in vitro properties. It is reasonable to assume that many compounds with poor pharmacokinetic properties are susceptible to oxidative metabolism. Existing in vitro liver microsome assays provide valuable information about this type of oxidative metabolism, which allows for the rational design of deuterated compounds of Formula I to enhance stability due to anti-oxidative metabolism. Consequently, the pharmacokinetic properties of compounds of Formula I are significantly improved, as quantitatively expressed by an extension of in vivo half-life (t/2), the concentration with the best efficacy ( _Cmax ), the area under the dose-response curve (AUC), and F, as well as by reduced clearance, dosage, and material cost.

The following description serves to illustrate the above: A series of analogs of a compound of formula I having multiple sites susceptible to oxidative metabolism, such as benzylic hydrogen atoms and nitrogen-bonded hydrogen atoms, were prepared. Various combinations of these hydrogen atoms were replaced with deuterium atoms, thereby replacing some, most, or all of the hydrogen atoms. Determination of the half-life advantageously and accurately determined the degree of improved resistance to oxidative metabolism. This method confirmed that this type of deuterium-hydrogen substitution could increase the half-life of the parent compound by up to 100%.

Deuterium-hydrogen substitution in compounds of Formula I can also be used to advantageously alter the metabolite profile of the starting compound to reduce or eliminate undesirable toxic metabolites. For example, if a toxic metabolite is produced by oxidative carbon-hydrogen (C-H) bond cleavage, it is reasonable to assume that a deuterated analogue will significantly reduce or eliminate the production of the undesirable metabolite, even if that particular oxidation reaction is not the rate-determining step. For more information on deuterium-hydrogen substitution in the prior art, see, for example, Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990, Reider et al., J. Org. Chem. 52, 3326-3334, 1987, Foster, Adv. Drug Res. 14, 1-40, 1985, Gillette et al., Biochemistry 33(10) 2927-2937, 1994, and Jarman et al. Carcinogenesis 16(4), 683-688, 1993.

As used herein, the term "modulator" is defined as a compound that binds to and/or inhibits a target with measurable affinity. In certain embodiments, the _IC50 and/or binding constant of the modulator is less than about 50 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM, or less than about 10 nM.

As used herein, the terms "measurable affinity" and "measurably inhibit" refer to a measurable change in FSHR activity between a sample containing a compound or composition of the invention and FSHR and an equivalent sample containing FSHR but without the compound or composition of the invention.

Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. As used herein, the term "stable" means possessing stability sufficient to allow manufacture and to maintain the integrity of the compound for a sufficient period of time to be useful for the various purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment of a variable herein includes that embodiment as any single embodiment or in combination with any other embodiment.

3. Description of Example Compounds

One aspect of the present invention provides a compound represented by formula I,

or a pharmaceutically acceptable salt, wherein:

X is O, S, SO, SO ₂ , or NR;

Y is O, S, or NR;

Z is O, S, SO, SO ₂ , or N; wherein when Z is O, S, SO, or SO ₂ , p is 0;

each R is independently hydrogen, a C _1-6 aliphatic group, a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; each of the above groups is optionally substituted; or

Two R on the same atom together with the atoms to which they are attached form a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; each of the above groups is optionally substituted;

Ring A is a fused C _3-10 aryl group, a fused 3-8 membered saturated or partially unsaturated carbocyclic ring, a fused 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or a fused 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur;

R ¹ is –OR, –SR, –CN, –NO ₂ , -SO ₂ R, -SOR, -C(O)R, -CO ₂ R, -C(O)N(R) ₂ , -NRC(O)R, -NRC(O)N(R) ₂ , -NRSO ₂ R, or –N(R) ₂ ;

R ² is –R, halogen, -haloalkyl, –OR, –SR, –CN, –NO ₂ , –SO ₂ R, –SOR, –C(O)R, –CO ₂ R, –C(O)N(R) ₂ , –NRC(O)R, –NRC(O)N(R) ₂ , –NRSO ₂ R, or –N(R) ₂ ;

R ³ is hydrogen, C _1-6 aliphatic, C _3-10 aryl, 3-8 membered saturated or partially unsaturated carbocyclic ring, 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; each of the above groups is optionally substituted;

each R ⁴ is independently —R, halogen, -haloalkyl, —OR, —SR, —CN, —NO ₂ , —SO ₂ R, —SOR, —C(O)R, —CO ₂ R, —C(O)N(R) ₂ , —NRC(O)R, —NRC(O)N(R) ₂ , —NRSO ₂ R, or —N(R) ₂ ;

R ⁵ is a C _1-6 aliphatic group, a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; each of the above groups is optionally substituted;

^R6 is hydrogen, _C1-6 aliphatic, _C3-10 aryl, 3-8 membered saturated or partially unsaturated carbocyclic ring, 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; each of the above groups is optionally substituted;

or R ⁵ and R ⁶ together with the atoms to which they are attached form a 3-8 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or a 3-8 membered heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; each of the above groups is optionally substituted;

n is 0, 1, or 2; and

p is 0 or 1.

In certain embodiments, X is O. In certain embodiments, X is S. In certain embodiments, X is SO or SO ₂ . In certain embodiments, X is NR.

In certain embodiments, Y is O. In certain embodiments, Y is S. In certain embodiments, Y is NR.

In certain embodiments, Z is O. In certain embodiments, Z is S. In certain embodiments, Z is SO or SO ₂ . In certain embodiments, Z is N.

In certain embodiments, Ring A is a fused C _3-10 aryl group. In certain embodiments, Ring A is a fused 3-8 membered saturated or partially unsaturated carbocyclic ring. In certain embodiments, Ring A is a fused 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In certain embodiments, Ring A is a fused 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In certain embodiments, Ring A is phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H indazolyl, indolenyl, isoxazolyl, morpholinyl, oxadiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, piperazinyl, piperidinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H pyrrolyl, pyrrolyl, tetrahydrofuranyl, thiazolyl, thienyl, thiophenyl, oxetanyl, or azetidinyl.

In certain embodiments, Ring A is phenyl.

In certain embodiments, R ¹ is —OR, —SR, —SO ₂ R, —SOR, —C(O)R, —CO ₂ R, —C(O)N(R) ₂ , —NRC(O)R, —NRC(O)N(R) ₂ , —NRSO ₂ R, or —N(R) _2. In certain embodiments, R ¹ is —OR, —SR, —SO ₂ R, or —SOR. In certain embodiments, R ¹ is —C(O)R, —CO ₂ R, or —C(O)N(R) _2. In certain embodiments, R ¹ is —NRC(O)R, —NRC(O)N(R) ₂ , —NRSO ₂ R, or —N(R) ₂ .

In certain embodiments, R ¹ is —OR, and R is hydrogen.

In certain embodiments, R ¹ is -OR, and R is a C _1-6 aliphatic group, a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the above groups is optionally substituted.

In certain embodiments, R ¹ is -OR, and R is a C _1-6 aliphatic group. In certain embodiments, R is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, a straight-chain or branched pentyl, or a straight-chain or branched hexyl; each of which is optionally substituted.

In certain embodiments, R ¹ is -OR, and R is C _3-10 aryl, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In certain embodiments, R is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecane, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzophenylthio, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, quinolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isoindoline, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindoline, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxazolyl 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, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4-[4-(2H-pyrrolidazole)] H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 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, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, R ² is hydrogen.

In certain embodiments, ^R is a _C1-6 aliphatic group. In certain embodiments, ^R is a _C1-6 aliphatic group, wherein the aliphatic group is a _C1-6 alkyl group. In certain embodiments, ^R is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, a straight-chain or branched pentyl group, or a straight-chain or branched hexyl group; each of which is optionally substituted. In certain embodiments, ^R is a _C1-6 aliphatic group, wherein the aliphatic group is a _C1-6 alkenyl group.

In certain embodiments, R ² is C _3-10 aryl, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the above groups is optionally substituted.

In certain embodiments, ^R is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecane, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, Carbolyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, dihydroindolinyl, indolizinyl, indolyl, 3H-indolyl, isoindoline, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindoline, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, 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, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizine thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, ^R2 is halogen, -haloalkyl, -OR, -SR, -CN, -NO2, _-SO2R , -SOR _, -C(O)R, _-CO2R , -C(O)N(R) ₂ , -NRC(O)R, -NRC(O)N(R) ₂ , _-NRSO2R , or -N(R) ₂ .

In certain embodiments, R ² is F, Cl, Br, I, or haloalkyl.

In certain embodiments, R ² is -OR, -SR, -CN, -NO ₂ , -SO ₂ R, -SOR, -C(O)R, -CO ₂ R, -C(O)N(R) ₂ , -NRC(O)R, -NRC(O)N(R) ₂ , -NRSO ₂ R, or -N(R) ₂ . In certain embodiments, R is a C _1-6 aliphatic group, a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the foregoing groups is optionally substituted. In certain embodiments, R is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, a straight-chain or branched pentyl group, or a straight-chain or branched hexyl group; each of the foregoing groups is optionally substituted. In other embodiments, R is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, acridinyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, Carbolyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolene, dihydroindolinyl, indolizinyl, indolyl, 3H-indolyl, isoindoline, isoindolineenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindoline, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, 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, benzoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoline oxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 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, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, R ² is hydrogen, Br, CN,

In certain embodiments, ^R2 is

In certain embodiments, ^R2 is

In certain embodiments, ^R2 is

In certain embodiments, R ³ is hydrogen.

In certain embodiments, R ³ is a C _1-6 aliphatic group, a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the above groups is optionally substituted.

In certain embodiments, ^R is an optionally substituted _C1-6 aliphatic group. In certain embodiments, ^R is an optionally substituted _C3-10 aryl group. In certain embodiments, ^R is an optionally substituted 3-8 membered saturated or partially unsaturated carbocyclic ring. In certain embodiments, ^R is an optionally substituted 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In certain embodiments, ^R is an optionally substituted 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In certain embodiments, ^R is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, acridinyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chroman yl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolene, dihydroindolinyl, indolizinyl, indolyl, 3H-indolyl, isoindoline, isoindolene, isobenzofuranyl, isochromanyl, isoindazolyl, isoindoline, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, 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, benzoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxaline phenyl, quinuclidine, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 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, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, ^R is dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolidinyl, imidazolyl, 1H-indazolyl, indazolyl, indolizinyl, indolyl, 3H-indolyl, isoindolinyl, isoindolinyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, purinyl, pyranyl, pyranyl, oxazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, tetrahydrofuranyl, 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, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, ^R3 is

In certain embodiments, each R ⁴ is independently hydrogen.

In certain embodiments, each R ⁴ is independently C _1-6 aliphatic, C _3-10 aryl, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the above groups is optionally substituted.

In certain embodiments, each R ⁴ is independently an optionally substituted C _1-6 aliphatic group. In certain embodiments, each R ⁴ is independently an optionally substituted C _3-10 aryl group. In certain embodiments, each R ⁴ is independently an optionally substituted 3-8 membered saturated or partially unsaturated carbocyclic ring. In certain embodiments, each R ⁴ is independently an optionally substituted 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In certain embodiments, each R ⁴ is independently an optionally substituted 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In certain embodiments, each R ⁴ is independently halogen, -haloalkyl, —OR, —SR, —CN, —NO ₂ , —SO ₂ R, —SOR, —C(O)R, —CO ₂ R, —C(O)N(R) ₂ , —NRC(O)R, —NRC(O)N(R) ₂ , —NRSO ₂ R, or —N(R) ₂ .

In certain embodiments, R ⁵ is a C _1-6 aliphatic group, a C _3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the above groups is optionally substituted.

In certain embodiments, ^R is an optionally substituted _C1-6 aliphatic group. In certain embodiments, ^R is an optionally substituted _C3-10 aryl group. In certain embodiments, ^R is an optionally substituted 3-8 membered saturated or partially unsaturated carbocyclic ring. In certain embodiments, ^R is an optionally substituted 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In certain embodiments, R is an optionally substituted 5-6 membered monocyclic heteroaromatic ring having ^1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In certain embodiments, R ⁵ is a C _1-6 aliphatic group. In certain embodiments, R ⁵ is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, a straight-chain or branched pentyl, or a straight-chain or branched hexyl; each of which is optionally substituted.

In certain embodiments, ^R is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecanyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, acridinyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chroman yl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolene, dihydroindolinyl, indolizinyl, indolyl, 3H-indolyl, isoindoline, isoindolene, isobenzofuranyl, isochromanyl, isoindazolyl, isoindoline, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, 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, benzoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxaline phenyl, quinuclidine, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 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, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, R ⁵ and R ⁶ together with the atoms to which they are each attached form a 3-8 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 3-8 membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of which is optionally substituted.

In certain embodiments, R ⁵ is methyl, tert-butyl, or -CD ₃ .

In certain embodiments, ^R5 is

In certain embodiments, Z is N, and the ring formed by Z, R ⁵ and R ⁶ is

In certain embodiments, R ⁶ is hydrogen.

In certain embodiments, ^R6 is a _C1-6 aliphatic group, a _C3-10 aryl group, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of the above groups is optionally substituted.

In certain embodiments, ^R is an optionally substituted _C1-6 aliphatic group. In certain embodiments, ^R is an optionally substituted _C3-10 aryl group. In certain embodiments, ^R is an optionally substituted 3-8 membered saturated or partially unsaturated carbocyclic ring. In certain embodiments, ^R is an optionally substituted 3-7 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In certain embodiments, ^R is an optionally substituted 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In certain embodiments, R ⁶ is a C _1-6 aliphatic group. In certain embodiments, R ⁶ is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, a straight-chain or branched pentyl, or a straight-chain or branched hexyl; each of which is optionally substituted.

In certain embodiments, R ⁶ is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, straight-chain or branched pentyl, or straight-chain or branched hexyl; each of which is optionally substituted.

In certain embodiments, ^R is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecane, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, acridinyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chroman yl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolene, dihydroindolinyl, indolizinyl, indolyl, 3H-indolyl, isoindoline, isoindolene, isobenzofuranyl, isochromanyl, isoindazolyl, isoindoline, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, 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, benzoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxaline phenyl, quinuclidine, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 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, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which is optionally substituted.

In certain embodiments, R ⁶ is hydrogen.

In certain embodiments, R ⁶ is methyl, tert-butyl, or -CD ₃ .

In certain embodiments, n is 0. In certain embodiments, n is 1. In certain embodiments, n is 2.

In certain embodiments, p is 0. In certain embodiments, p is 1.

In certain embodiments, ^R1 , ^R2 , ^R3 , ^R4 , ^R5 , ^R6 , X, Y, Z, n, and p are each as defined above and further described by embodiment, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides a compound represented by formula I-a,

or ^a pharmaceutically acceptable salt thereof, wherein ^R1 , ^R2 , R3, ^R4 , ^R5 , ^R6 , X, Y, Z, and p are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides a compound represented by formula I-b,

or ^a pharmaceutically acceptable salt thereof, wherein ^R1 , ^R2 , R3, ^R4 , ^R5 , ^R6 , Y, Z, n, and p are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides a compound represented by formula I-c,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ , R ⁵ , R ⁶ , Z, and p are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides a compound represented by formula I-d,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ , R ⁵ , and R ⁶ are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides a compound represented by formula I-e,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ , R ⁴ , and R ⁵ are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In other embodiments, the present invention provides a compound represented by formula I-f,

or a pharmaceutically acceptable salt thereof, wherein R ² , R ³ , R ⁵ , and R ⁶ are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In other embodiments, the present invention provides a compound represented by formula I-g,

or a pharmaceutically acceptable salt thereof, wherein R ² , R ³ , and R ⁵ are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides compounds represented by formula I-h,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ⁵ , R ⁶ , Z, and p are each as defined above and further described by example, class, subclass, and herein, alone or in combination.

In certain embodiments, the present invention provides compounds of formula If, wherein ^R2 is a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; ^R3 is an optionally substituted 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; and Z is N.

In certain embodiments, R ⁵ is an optionally substituted C _1-6 aliphatic group.

In certain embodiments, R ⁵ and R ⁶ , together with the atoms to which they are each attached, form a 3-8 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which is optionally substituted.

In certain embodiments, R ⁶ is an optionally substituted C _1-6 aliphatic group.

In certain embodiments, the present invention provides a compound of formula Ih, wherein R ¹ is -OR, and R is a C _1-6 aliphatic group; R ² is a 5-6 membered monocyclic heteroaryl group having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein the heterocyclic ring is optionally substituted; and Z is N.

In certain embodiments, R ⁵ is an optionally substituted C _1-6 aliphatic group.

In certain embodiments, R ⁵ and R ⁶ , together with the atoms to which they are each attached, form a 3-8 membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which is optionally substituted.

In certain embodiments, R ⁶ is an optionally substituted C _1-6 aliphatic group.

In certain embodiments, the present invention provides compounds given in Table 1.

Table 1

In some embodiments, the present invention provides a compound selected from the compounds described above or a pharmaceutically acceptable salt thereof.

Various structural representations may show heteroatoms without groups, radicals, charges, or counterions attached thereto. One of ordinary skill in the art will understand that such representations mean that the heteroatoms are attached to hydrogen (eg, should be understood as).

In certain embodiments, the compounds of the present invention are synthesized according to the following Schemes A-C. More specific examples of compounds prepared using Schemes A-C are provided in the Examples below.

Process A

Process B

Process C

4. Use, preparation and administration

Pharmaceutically acceptable compositions

According to another embodiment, the present invention provides a composition comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of the compound in the composition of the present invention is effective to measurably modulate FSHR or a variant thereof in a biological sample or patient. In certain embodiments, the amount of the compound in the composition of the present invention is effective to measurably modulate FSHR or a variant thereof in a biological sample or patient. In certain embodiments, the composition of the present invention is formulated for administration to a patient in need thereof.

As used herein, the term "patient" or "subject" refers to an animal, preferably a mammal, and most preferably a human.

The term "pharmaceutically acceptable carrier, adjuvant or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound formulated therewith. Pharmaceutically acceptable carriers, adjuvants, or vehicles used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block copolymers, polyethylene glycol, and lanolin.

"Pharmaceutically acceptable derivative" refers to any non-toxic salt, ester, ester salt or other derivative of a compound of the invention which, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the invention or a metabolite or residue having inhibitory activity.

The compositions of the present invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, orally, vaginally, or by implantation into a container. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions of the present invention include aqueous or oily suspensions. These suspensions are prepared using suitable dispersing or wetting agents and suspending agents according to techniques known in the art. Sterile injectable preparations can also be sterile injectable solutions or suspensions made with nontoxic parenteral acceptable diluents or solvents (e.g., solutions in 1,3-butanediol). Acceptable vehicles and solvents used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are typically used as solvents or suspending media.

For this reason, any available gentle fixed oil comprises synthetic mono- or di-glyceride. Fatty acids such as oleic acid and glyceride derivatives thereof can be used to prepare injection, as natural pharmaceutically acceptable oils, for example olive oil or castor oil, especially their polyoxyethylated oils. These oil solutions or suspensions also contain long-chain alcohol diluents or dispersants, for example carboxymethyl cellulose or similar dispersants, which are usually used in the preparation of pharmaceutically acceptable dosage forms, including emulsions and suspensoids. Other conventional surfactants, such as Tweens, Spans and other emulsifiers or bioavailability enhancers, are usually used in the preparation of pharmaceutically acceptable solids, liquids, or other dosage forms also can be used for the purpose of preparation.

The pharmaceutically acceptable compositions of the present invention are administered orally in any orally acceptable dosage form. Exemplary oral dosage forms are capsules, tablets, aqueous suspensions, or solutions. For oral tablets, common carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. For oral administration in capsule form, useful diluents include lactose and dry corn starch. When an oral aqueous suspension is desired, the active ingredient is combined with an emulsifier and a suspending agent. Optionally, certain sweeteners, flavorings, or coloring agents may be added, if desired.

Alternatively, the pharmaceutically acceptable compositions of the present invention are administered in the form of suppositories for rectal administration. These are prepared by mixing the agent with a suitable non-irritating excipient, which is solid at room temperature but liquid at rectal temperature and will therefore melt in the rectum and release the drug. Such materials include cocoa butter, beeswax, and propylene glycol.

The pharmaceutically acceptable compositions of this invention can also be administered topically, especially when the target of treatment includes areas and organs readily accessible by topical application, including diseases of the eye, skin, or lower intestinal tract. Suitable topical formulations are readily prepared for each area or organ.

Topical application to the lower intestinal tract can be achieved in a rectal suppository formulation (see above) or in a suitable enema formulation.Topical transdermal patches may also be used.

For topical application, a pharmaceutically acceptable composition is prepared in a suitable ointment containing an active ingredient suspended or dissolved in one or more carriers. Exemplary carriers for topical application of the compounds of the present invention include mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene olefins, polyoxypropylene olefin compounds, emulsifying wax and water. Alternatively, a pharmaceutically acceptable composition is prepared in a suitable lotion or cream containing an active ingredient suspended or dissolved in one or more carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, CETE aryl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The pharmaceutically acceptable compositions of the present invention are optionally administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and can be prepared as solutions in saline, using benzyl alcohol or other suitable preservatives and absorption enhancers to enhance bioavailability, and can also use fluorocarbons and/or other conventional solubilizing agents or dispersants.

Most preferably, the pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present invention are not administered with food. In other embodiments, the pharmaceutically acceptable compositions of the present invention are administered with food.

The compounds of the present invention are optionally combined with carrier materials to produce a single dosage form of the composition, the amount of which will depend on the host being treated, the specific mode of administration. Preferably, the compositions provided should be formulated into a dosage between 0.01-100 mg/kg body weight/compound, which can be administered daily to patients receiving these compositions.

It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound employed, age, weight, general health, sex, dietary status, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the specific disease. The amount of the compound of the invention in the composition will also depend on the specific compound in the composition.

Uses of compounds and pharmaceutically acceptable compositions

In certain embodiments, the present invention provides a method for agonizing FSHR or a mutant thereof in a positive allosteric manner in a patient or biological sample, comprising administering to the patient a compound of the present invention or contacting the biological sample with a compound of the present invention.

In certain embodiments, the present invention relates to the use of compounds of the present invention and/or physiologically acceptable salts thereof for modulating the FSH receptor (particularly in the presence of FSH). The term "modulate" refers to any change in FSHR-mediated signaling resulting from the action of a specific compound of the present invention, which is capable of interacting with the FSHR target, enabling recognition, binding, and activation. Compounds are characterized by high affinity for the FSHR, ensuring reliable binding, preferably positive allosteric modulation of the FSHR. In certain embodiments, the substance is monospecific, ensuring unique and direct recognition of a single FSHR target. In the context of the present invention, the term "recognition"—without limitation—refers to any type of interaction between a specific compound and a target, particularly covalent or non-covalent binding or association, such as covalent bonds, hydrophilic/hydrophobic interactions, van der Waals forces, ion pairs, hydrogen bonds, ligand-receptor interactions, and the like. Such association may also include the presence of other molecules (e.g., peptides, proteins, or nucleotide sequences). The receptor/ligand interactions of the present invention are characterized by high affinity, high selectivity and minimal to no cross-reactivity with other target molecules, thereby precluding unhealthy and deleterious effects on the treated subject.

In certain embodiments, the present invention relates to a method for regulating the FSH receptor, preferably in a positive allosteric manner, wherein a system capable of expressing the FSH receptor is contacted with at least one compound of Formula I of the present invention and/or a physiologically acceptable salt under conditions that regulate the FSH receptor in the presence of FSH. In certain embodiments, the system is a cell system. In other embodiments, the system is an in vitro translation system, which is based on protein synthesis and does not require living cells. A cell system is defined as any subject, as long as the subject contains cells. Therefore, the cell system can be selected from a single cell, a cell culture medium, a tissue, an organ, and an animal. In certain embodiments, the method for regulating the FSH receptor is performed in vitro. The teachings of the above description regarding compounds of Formula I (including any preferred embodiments thereof) are effective and applicable, and are not limited to compounds of Formula I and their salts in methods for regulating FSHR.

In certain embodiments, the compounds of the present invention exhibit favorable biological activity that is readily demonstrated in cell culture assays, such as those described herein or in the prior art (see, for example, WO 2002/09706, the contents of which are incorporated herein by reference). In such assays, the compounds of the present invention preferably exhibit and elicit an agonist effect. In certain embodiments, the compounds of the present invention have FSHR agonist activity, expressed as an EC ₅₀ of less than ₅ μM. In certain embodiments, the EC ₅₀ is less than 1 μM. In certain embodiments, the EC 50 is less than 0.5 μM. In certain embodiments, _{the EC 50 is} less than 0.1 μM. "EC ₅₀ " is the effective concentration of a compound that achieves half the maximal response of FSH.

As discussed herein, these signaling pathways are associated with various diseases, including fertility disorders. Conditions/diseases treated by the methods of the present invention include, but are not limited to, hypogonadism, isolated idiopathic hypogonadism, Kallmann syndrome, idiopathic hypogonadism, craniopharyngioma, combined pituitary hormone deficiency, fertile anorchia syndrome, abnormal LH beta subunit, abnormal FSH beta subunit, masses, pituitary adenomas, cysts, metastatic cancer to the sella (breast in women, lung and prostate in men), infiltrative lesions, hemochromatosis, sarcoidosis, histiocytosis, lymphoma, lymphocytic hypophysitis, infection, meningitis, pituitary apoplexy, hyperprolactinemia, hypothyroidism, intentional (iatrogenic) secondary hypogonadism, empty sella turcica, pituitary infarction, Sheehan syndrome, anorexia nervosa, congenital adrenal hyperplasia, and conditions associated with GnRH deficiency. Therefore, the compounds of the present invention are made to prevent and/or treat diseases that depend on the signal transduction pathway by interacting with one or more of the signal transduction pathways. Therefore, the present invention relates to compounds according to the present invention as modulators of the signal transduction pathways (preferably FSHR-mediated signal transduction pathways) described in this specification, preferably agonists, more preferably positive allosteric modulators. It is generally believed that the compounds of the present invention are able to bind to the intracellular receptor domain and have no competitive interaction with FSH, but they act on their receptors as positive allosteric enhancers of FSH. Non-competitive interaction refers to the agonist activity properties exhibited by the compounds of the present invention, wherein the compounds activate FSHR without significantly reducing the binding strength of FSH to FSHR.

In certain embodiments, the present invention is directed to stimulation of follicular development, ovulation induction, controlled ovarian hyperstimulation, assisted reproductive technology (including in vitro fertilization), male hypogonadism, and male infertility, including certain conditions where sperm production is inadequate.

Another object of the present invention is to provide methods for treating diseases caused, mediated, and/or propagated by FSHR activity, wherein at least one compound of Formula I and/or a physiologically acceptable salt thereof is administered to a mammal in need of such treatment. In certain embodiments, the present invention provides methods for treating fertility disorders, wherein at least one compound of Formula I and/or a physiologically acceptable salt thereof is administered to a mammal in need of such treatment. In certain embodiments, an effective amount of the compound is administered as described above. In certain embodiments, the treatment is administered orally.

In certain embodiments, the treatment method is to achieve ovulation induction and/or controlled superovulation. In another preferred aspect, the treatment method forms the basis of an in vitro fertilization method, comprising the steps of: (a) treating a mammal with the treatment method described above, (b) collecting eggs from the mammal, (c) fertilizing the eggs, and d) implanting the fertilized eggs in a host mammal. The host mammal can be a mammal undergoing treatment (i.e., a patient) or a surrogate mother. The teachings of the present invention and its embodiments are valid and applicable and, if it seems reasonable, are not limited to the treatment methods of the present invention.

The methods of the present invention can be performed in vitro or in vivo. In vitro assays can be used to determine the sensitivity of specific cells to treatment with the compounds of the present invention during research and clinical applications. Cell cultures are typically incubated with various concentrations of the compounds of the present invention for a sufficient time to allow the active agent to modulate FSHR activity, typically between one hour and one week. In vitro treatment can be performed using cultured cells from biopsy samples or cell lines. In a preferred aspect of the present invention, maturation of follicular cells is stimulated. Viable cells remaining after treatment are then counted and further processed.

The host or patient can be a mammal, such as a primate, particularly a human; a rodent, including mice, rats, and hamsters; a rabbit, a horse, a cow, a dog, a cat, etc. Animal models of interest for experimental research can provide models for treating human diseases.

To identify signal transduction pathways and to detect interactions between various signal transduction pathways, many scientists have developed appropriate models or model systems, such as cell culture models and transgenic animal models. To detect certain stages of the signal transduction cascade, interacting compounds can be used to modulate the signal. The compounds of the present invention can be used as reagents to detect FSHR-dependent signal transduction pathways in animal models and/or cell culture models or in clinical diseases described in this application.

The uses described in the above paragraphs of this specification can be carried out in vitro or in vivo models. The regulation is monitored by the techniques described in the context of this specification. In certain embodiments, the in vitro use is used on human samples suffering from fertility disorders. By testing a variety of specific compounds and/or their derivatives, the active ingredient most suitable for the treatment of human patients can be selected. The in vivo dose rate of the selected derivative should be matched with the susceptibility to FSHR and/or the severity of the patient's disease based on in vitro (test) data, thereby significantly improving the therapeutic effect. In addition, the following teachings of this specification concerning the use of compounds of formula (1) and their derivatives in the preparation of drugs for preventing, treating and/or controlling the course of the disease are valid and applicable, and if it seems reasonable, may not be limited to the use of the compounds of the present invention to regulate FSHR activity.

The present invention also relates to the use of compounds of Formula I and/or physiologically acceptable salts thereof in the preventive or therapeutic treatment and/or monitoring of diseases caused, mediated, and/or propagated by FSHR activity. Furthermore, the present invention relates to the use of compounds of Formula I and/or physiologically acceptable salts thereof in the preparation of medicaments for the preventive or therapeutic treatment and/or monitoring of diseases caused, mediated, and/or propagated by FSHR activity. In certain embodiments, the present invention provides the use of compounds of Formula I and/or physiologically acceptable salts thereof in the preparation of medicaments for the preventive or therapeutic treatment of diseases mediated by FSHR.

In addition, the compounds of formula I and/or their physiologically acceptable salts can be used as intermediates for the preparation of other active pharmaceutical ingredients. The pharmaceuticals are preferably prepared by non-chemical methods, for example by combining the active ingredient with at least one solid, liquid and/or semi-liquid carrier or excipient, optionally admixed with one or more other active substances in a suitable dosage form.

Another object of the present invention is to use the compounds of formula I of the present invention and/or their physiologically acceptable salts for the preventive or therapeutic treatment and/or monitoring of diseases caused, mediated and/or spread by FSHR activity. Another preferred object of the present invention relates to the compounds of formula I of the present invention and/or their physiologically acceptable salts for the preventive or therapeutic treatment and/or monitoring of fertility disorders. In addition, the teachings of this specification regarding the compounds of formula (1) (including any preferred embodiments thereof) are valid and applicable, and may not be limited to the use of the compounds of formula (1) and their salts for the preventive or therapeutic treatment and/or monitoring of fertility disorders.

The compound shown in the general formula I of the present invention can be administered once or multiple times as treatment before or after the onset of the disease. The above-mentioned compounds and medical products of the present invention are particularly used for therapeutic treatment. The therapeutic effect refers to the relief of one or more disease symptoms to a certain extent, or the partial or complete reversal of one or more physiological or biochemical parameters related to the disease or pathology or causing disease or pathological changes to a normal state. If the compound is administered at different time intervals, monitoring can be considered as a treatment to enhance the response and completely eradicate the disease pathogens and/or symptoms. The same or different compounds can be applied. Drugs can also be used to reduce the development of the disease or even prevent the occurrence of symptoms related to FSHR activity in advance, or to treat existing or persistent symptoms. The disease or condition to which the present invention relates is preferably a fertility disorder.

In the sense of the present invention, if the subject has pre-existing conditions for the above physiological or pathological conditions, such as a familial predisposition, a genetic defect, or a past history of the disease, it is advisable to administer a preventive treatment.

The present invention also relates to a medicament containing at least one compound of the present invention and/or its pharmaceutically acceptable derivatives, salts, solvates and stereoisomers, including mixtures thereof in various ratios. In certain embodiments, the present invention also relates to a medicament containing at least one compound of the present invention and/or its physiologically acceptable salts.

A "drug" within the meaning of the present invention is any agent from the pharmaceutical field, including one or more compounds of the general formula I or their preparations (e.g. pharmaceutical compositions or pharmaceutical preparations), which can be used for the prevention, treatment, follow-up or post-treatment care of patients suffering from diseases associated with FSHR activity, said patients at least temporarily showing pathological changes in the overall condition or in individual parts of the patient's body.

In various embodiments, the active ingredient can be administered alone or in combination with other therapies. Synergistic effects can be achieved by using one or more compounds according to the present invention in pharmaceutical compositions, i.e., combining a compound of Formula I with at least one other active agent, which can be another compound of Formula I or a compound with a different structural framework. These active ingredients can be administered simultaneously or sequentially. The compounds of the present invention are suitable for combination with known fertility inducing agents. Preferably, the additional active agent is selected from the group consisting of FSH, α-FSH (Gonal F), β-FSH, LH, hMG, and 2-(4-(2-chloro-1,2-diphenylvinyl)-phenoxy)-N,N-diethyl-citrate ethylamine (clomiphene citrate). Other ovulation additives are known to those skilled in the art (see, for example, WO 2002/09706, the contents of which are incorporated herein by reference) and can be used with the compounds of the present invention.

Another aspect of the present invention provides a kit comprising separately packaged effective amounts of a compound of the present invention and/or its pharmaceutically acceptable salts, derivatives, solvates, and stereoisomers, including mixtures thereof in various ratios, and optionally an effective amount of other active ingredients. The kit comprises a suitable container, for example, a box, various bottles, bags, or ampoules. For example, the kit may comprise separate ampoules, each containing an effective amount of a compound of the present invention and/or its pharmaceutically acceptable salts, derivatives, solvates, and stereoisomers, including mixtures thereof in various ratios, in dissolved or lyophilized form, and an effective amount of other active compounds.

As used herein, the term "treat" refers to reversing, slowing down, delaying the appearance of a disease or condition described herein or one or more symptoms, or inhibiting the development of a disease or condition described herein or one or more symptoms. In some embodiments, treatment is given after one or more symptoms have appeared. In other embodiments, treatment is given in the absence of symptoms. For example, treatment is given before the onset of symptoms in a susceptible individual (e.g., based on symptom history and/or genetic or other sensory factors). Treatment can be continued after symptoms disappear, for example to prevent or delay their recurrence.

The compounds and compositions according to the methods of the present invention are administered in any dosage amount and by any route of administration that is effective in treating or alleviating the severity of the aforementioned conditions. The exact amount required will vary from subject to subject, depending on the race, age, and general condition of the subject, the severity of the infection, the specific drug, the route of administration, and the like. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, the term "unit dosage form" refers to a physically discrete unit suitable for administering a single dose to the patient to be treated. However, it will be understood that the total daily dosage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific dosage level for any particular patient or organism will depend on a variety of factors, including the specific condition to be treated and its severity, the activity of the specific compound selected, the specific composition used, age, weight, health status, sex, dietary status, time and route of administration, the excretion rate of the specific compound, the duration of treatment, drugs used in conjunction with or in combination with the specific compound, and other factors well known in the art.

The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternal, intravaginal, intraperitoneally, topically (e.g., by powder, ointment, or drops), buccally, orally or nasally by spray, etc., depending on the severity of the infection. In certain embodiments, the compounds of the present invention are administered orally or parenterally at a dosage level of about 0.01 mg/kg to 100 mg/kg of subject weight, preferably about 1 mg/kg to 50 mg/kg of subject weight, once or more daily to achieve the desired therapeutic effect.

Liquid dosage forms suitable for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form optionally contains inert diluents commonly used in the art (e.g., water or other solvents), solubilizers and emulsifiers (e.g., ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyl ethylene glycol, 1,3-butyl ethylene glycol, dimethylformamide, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings and aromatics.

Injectable preparations, such as sterile injectable aqueous or oily suspensions, are prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be prepared as sterile injectable solutions, suspensions or emulsions using non-toxic parenteral acceptable diluents or solvents (e.g., solutions in 1,3-butanediol). Among the acceptable carriers and solvents that can be used are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, non-volatile oils are generally used as oil solvents or suspending media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can be used to prepare injections.

Prior to use, the injectable preparations are sterilized by passing through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable media.

In order to prolong the effect of the compounds of the present invention, the absorption of the compounds by subcutaneous or intramuscular injection is generally slowed. This can be achieved by using a liquid suspension of crystalline or amorphous materials with poor water solubility. The absorption rate of the compound depends on its dissolution rate, which in turn varies depending on the crystal size and crystalline form. Alternatively, the absorption of the compound administered parenterally can be delayed by dissolving or suspending the compound in an oil carrier. A microcapsule matrix of the compound is formed in a biodegradable polymer (polylactide-polyglycolide) to prepare a reservoir form of injection. Depending on the ratio of the compound to the polymer and the properties of the specific polymer used, the release rate of the compound can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot-type injectable preparations can also be prepared by embedding the compound in liposomes or microemulsions compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, wherein the excipients are solid at room temperature but liquid at body temperature and therefore will melt in the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier, which includes sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starch, lactose, sucrose, glucose, mannose, and silicic acid, b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and gum arabic, c) humectants such as glycerol, d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retardants such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets or pills, the dosage form may also contain buffering agents.

The solid composition of similar type can also be used as filler for soft or hard filled gelatin capsules, and described capsule uses for example lactose or milk sugar, and the polyethylene glycol of high molecular weight etc. as excipient.The solid dosage form of tablet, dragee, capsule, pill and granule can adopt coating and shell preparation, for example other coating known in the field of enteric coating and pharmaceutical preparation.They optionally contain opacifier, and become composition, optionally in a delayed manner, only (or preferably) release active ingredient in the specific position of intestinal tract.The example of spendable embedding composition comprises polymer and wax.The solid composition of similar type can also be used as filler for soft or hard filled gelatin capsules, and described capsule uses for example lactose or milk sugar, and the polyethylene glycol of high molecular weight etc. as excipient.

The active compound can also be formulated into a microcapsule form together with one or more excipients, such as the excipients described above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared using coatings and shells, such as enteric coatings, controlled release coatings and other coatings known in the field of pharmaceutical formulation. In the solid dosage form, the active compound is mixed with at least one inert diluent, such as sucrose, lactose or starch. In normal practice, such dosage forms also include tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose, in addition to the inert diluent. In the case of capsules, tablets or pills, a buffer can also be contained in the dosage form. They optionally contain an opacifying agent and become a composition, optionally in a delayed manner, only (or preferably) releasing the active ingredient in a specific part of the intestinal tract. Examples of usable embedding compositions include polymers and waxes.

The dosage forms of the compounds of the present invention for topical or transdermal administration include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active compound is mixed with a pharmaceutically acceptable carrier and any required preservatives, buffers or propellants under sterile conditions. Ophthalmic preparations, ear drops and eye drops are also within the scope of the present invention. In addition, the present invention encompasses the use of transdermal patches, which have the additional advantage of being able to controllably deliver the compound to the body. Such dosage forms can be prepared by dissolving or dispersing the compound in an appropriate medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. Providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel can control the rate.

According to one embodiment, the present invention relates to a method for allosterically modulating FSHR activity in a biological sample, said method comprising the step of contacting said biological sample with a compound of the present invention or a composition comprising a compound of the present invention.

According to another embodiment, the present invention relates to a method for modulating the activity of FSHR or a mutant thereof in a positive allosteric manner in a biological sample, comprising the step of contacting the biological sample with a compound of the present invention or a composition comprising the compound of the present invention.

The compounds of the present invention are potent and selective modulators of the FSH receptor. They are 3-10 times more selective for the FSH receptor than for the LH receptor, and even 10-100 times more selective for the TSH receptor, while having _EC50 or _IC50 values of no more than 10 μM for unrelated G protein-bound receptors (GPCRs) or non-GPCR targets. The present invention includes the use of the compounds of the present invention to regulate and/or modulate the signaling cascade of the FSHR, thereby providing research tools for the diagnosis and/or treatment of any disease caused by FSHR signaling.

For example, the compounds of the present invention are used in vitro as unique tools for understanding the biological effects of FSH, which include evaluating many factors that are thought to affect FSH production and the interaction between FSH and FSHR (e.g., the mechanism of FSH signal transduction/receptor activation), as well as many factors that are affected by FSH production and the interaction between FSH and FSHR. The compounds of the present invention can also be used to develop other compounds that interact with FSHR because the compounds of the present invention provide important structure-activity relationship (SAR) information for such development. The compounds of the present invention bind to FSHR and can be used as reagents for detecting FSHR on living cells, fixed cells, in biological fluids, in tissue homogenates, in pure natural materials, etc. For example, by labeling the compounds, one can identify cells with FSHR on their surface. In addition, because the compounds of the present invention have the ability to bind to FSHR, they can be used in in situ staining, FACS (fluorescence activated cell sorting), Western blotting, ELISA (enzyme-linked immunosorbent assay), receptor purification, or for purifying cells that express FSHR on the cell surface or in permeabilized cells.

The compounds of the present invention may also be used as commercial research reagents for various medical research and diagnostic applications. Such uses include, but are not limited to: use as calibration standards for quantifying the activity of candidate FSH agonists in various functional assays; use as blocking agents in random compound screening, i.e., compounds can be used to block the activation of the FSH compounds claimed herein when searching for a new family of FSH receptor ligands; use in co-crystallization with the FSHR receptor, i.e., compounds of the present invention allow the compound to bind to the FSHR to form crystals, thereby determining the receptor/compound structure by X-ray crystallography; use in other research and diagnostic applications where FSHR activation is preferred or such activation can be conveniently calibrated relative to a known amount of an FSH agonist; use as probes in assays to determine FSHR expression on the cell surface; and use in the development of compounds for detecting sites where FSHR binding ligands are located in assays.

The compounds of the present invention can be administered or combined with physical means to diagnose therapeutic effects. Pharmaceutical compositions containing the compounds and their use in treating FSHR-mediated conditions represent a promising new approach with potential applications in a variety of therapeutic settings, enabling direct and immediate improvements in human or animal health. Their effectiveness is particularly beneficial for effectively treating infertility, whether used alone or in conjunction with other fertility-inducing treatments. In particular, the compounds of the present invention enhance the effects of natural FSH in ovulation induction and assisted reproductive technology. The oral bioavailability and novel active chemical substance of the present invention further facilitate patient and physician compliance.

The compounds of the present invention were active in the primary screen (CHO cells with or without FSHR), selective in the secondary screen (no or low activity against TSHR and LHR), and potent in the estradiol enzyme assay for granulosa cells. No hERG or any toxic effects were observed in vitro.

In certain embodiments, the present invention provides a method of in vitro fertilization, comprising the steps of:

(a) treating a mammal according to the method above,

(b) collecting eggs from said mammal,

(c) fertilizing the egg, and

(d) implanting the fertilized egg into a host mammal.

The compounds of formula I and their salts, isomers, tautomers, enantiomeric forms, diastereomers, racemates, derivatives, prodrugs and/or metabolites are characterized by high specificity and stability, low manufacturing costs and convenient handling. These characteristics form the basis for their reproducible action, including the absence of cross-reactivity, and reliable and safe interaction with the target structure.

As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof, biopsy material obtained from a mammal or extracts thereof, and blood, saliva, urine, feces, semen, tears or other body fluids or extracts thereof.

Modulating the activity of FSHR or its mutants in a biological sample can be used for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ transplantation, biological sample storage, and biological testing.

DETAILED DESCRIPTION

As described in the Examples below, in certain exemplary embodiments, compounds were prepared according to the following general procedures. It should be understood that although the general methods describe the synthesis of certain compounds of the present invention, the following general methods, as well as other methods known to those of ordinary skill in the art, are also applicable to the synthesis of all compounds described herein and subclasses and species of each compound.

The compound numbers used in the following examples correspond to the compound numbers above.

^1H NMR (Nuclear Magnetic Resonance) ^1H NMR spectrometers were recorded on a Bruker 400 MHz spectrometer using the residual signal of the deuterated solvent as an internal standard. Chemical shifts were reported relative to tetramethylsilane and expressed in parts per million (ppm). ^1H NMR data were reported as follows: chemical shift (multiplicity, coupling constant, and number of hydrogen atoms). Multiplicities were expressed as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad).

LCMS analysis was performed under the following conditions:

Method A: 0.1% TFA in water, B: 0.1% TFA in ACN

Run time: 6.5 minutes

Flow rate: 1.0 mL/min

Gradient: 5-95% B, 4.5 min, wavelength: 254 and 215 nM.

Column: Waters Sunfire C18, 3.0x50mm, 3.5um forward mode

Mass scan: 100-900Da

Process 1:

Example 1

7-Methoxy-8-(1H-pyrazol-4-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (1)

Step 1: 3-Bromo-1-(2,4-dihydroxyphenyl)propan-1-one

Trifluoromethanesulfonic acid (75 mL, 0.84 mol) was added dropwise to resorcinol (25 g, 0.22 mol) and 3-bromopropionic acid (38.3 g, 0.25 mol) at 0°C under a nitrogen atmosphere with stirring. After the addition was complete, the reaction mixture was heated at 80°C for 30 minutes. The reaction mixture was cooled to room temperature, quenched with ice water (200 mL), and extracted with DCM (500 mL). The aqueous phase was further extracted with DCM (2 x 100 mL); the organic phases were combined, dried over sodium sulfate, and concentrated in vacuo to afford the desired compound (40 g, 72%) as an orange solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ12.2 (bs, 1H), 10.6 (bs, 1H), 7.77-7.75 (d, J=8Hz, 1H), 6.38-6.35 (dd, J=2.0, 8.8Hz, 1H ), 6.26-6.26 (d, J=4.0Hz, 1H), 3.74 (t, J=6.8Hz, 2H), 3.61-3.57 (dd, J=6.0, 11.2Hz, 2H).

Step 2: 7-Hydroxy-2,3-dihydro-4H-benzopyran-4-one

To an ice-cold solution of 2M NaOH (92 mL, 2.33 mol) was added 3-bromo-1-(2,4-dihydroxyphenyl)propan-1-one (38 g, 0.155 mol) portionwise over 30 minutes. The resulting suspension was stirred at room temperature for 2 hours. The reaction mixture was cooled to 0°C; the pH was adjusted to approximately 2 with 50% aqueous sulfuric acid. The solid was filtered, stirred at room temperature for 10 minutes longer, filtered, and dried under high vacuum to yield the desired compound (16 g, 63%) as a tan solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ10.52 (bs, 1H), 7.61-7.59 (d, J = 8.0Hz, 1H), 6.48-6.45 (dd, J = 4.0, 12.0Hz, 1H), 6.29-6.20 (d, J=4.0Hz, 1H), 4.44 (t, J=4.0Hz, 2H), 2.65 (t, J=4.0Hz, 2H).

Step 3: 7-Methoxy-2,3-dihydro-4H-chromen-4-one

To a solution of 7-hydroxy-2,3-dihydro-4H-chromen-4-one (27 g, 0.16 mol) in acetone (700 mL) was added portionwise _{dry K₂CO₃} (45.6 g, 0.32 mol) with stirring at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 10 minutes, followed by the dropwise addition of iodomethane (65.4 g, 0.46 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was filtered and concentrated in vacuo. The crude product was dissolved in DCM (200 mL), washed with water (100 mL), brine (50 mL), dried over sodium sulfate, and concentrated in vacuo to afford the desired compound (15 g, 89%) as a light yellow solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.85-7.83 (d, J=8.0Hz, 1H), 6.60-6.57 (dd, J=4.0, 12.0Hz, 1H), 6.41-6.4 1 (d, J=4.0Hz, 1H), 4.52 (t, J=8.0Hz, 2H), 3.77 (s, 3H), 2.76 (t, J=4.0Hz, 2H).

Step 4: 6-Bromo-7-methoxy-2,3-dihydro-4H-chromen-4-one

To a solution of 7-methoxy-2,3-dihydro-4H-chromen-4-one (30 g, 0.16 mol) in a mixture of acetonitrile and diethyl ether (100:300 mL) was added portionwise silica gel 60-120 mesh (1.5 g) and NBS (33 g, 0.18 mol) at room temperature under nitrogen. The reaction mixture was stirred at room temperature for 14 hours. The reaction mixture was filtered and concentrated in vacuo. The crude product was purified by column chromatography using petroleum ether/ethyl acetate (9:1) as eluent to afford the desired compound (10 g, 72%) as a light brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.82 (s, 1H), 6.73 (s, 1H), 4.53 (t, J=8.0Hz, 2H), 3.89 (s, 3H), 2.72 (t, J=8.0Hz, 2H).

Step 5: Ethyl (6-bromo-7-methoxy-4-oxo-2H-chromen-3(4H)-ylidene)(hydroxy)acetate

Diisopropylamine (7 mL, 0.3958 mol) was dissolved in dry THF (50 mL) at room temperature under a nitrogen atmosphere. The reaction mixture was cooled to -78°C, and n-butyllithium (1.6 M solution in hexane, 29.2 mL, 0.04 mol) was added dropwise over 30 minutes. After the addition was complete, the reaction mixture was stirred at the same temperature, then slowly warmed to -10°C and stirred for 30 more minutes. The reaction mixture was cooled again to -78°C, and a solution of 6-bromo-7-methoxy-2,3-dihydro-4H-chromen-4-one (10 g, 0.03 mol) in THF (50 mL) was added dropwise over 30 minutes, followed by stirring at -78°C. After 1 hour, diethyl oxalate (7.8 mL, 0.05 mol) was added dropwise at -78°C; the reaction mixture was slowly warmed to 0°C and stirred for 1 hour. The reaction mixture was cooled to -5°C and quenched by the addition of 1.5N HCl solution, and extracted with ethyl acetate (100 mL x 2). The organic layers were combined, washed with water (100 mL), brine (50 mL), dried over sodium sulfate, and concentrated to give the desired compound (10 g, 72%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ.8.04 (s, 1H), 6.43 (s, 1H), 5.32 (s, 2H), 4.40-4.32 (m, 2H), 3.95 (s, 3H), 1.42-1.37 (m, 3H).

Step 6: tert-Butyl 1-(3-thienyl)hydrazinecarboxylate

To a solution of 3-bromothiophene (10 g, 0.061 mol) in DMSO (100 mL) at room temperature under nitrogen was added tert-butyl hydrazinocarboxylate (16.3 g, 0.122 mol), cesium carbonate (40 g, 0.122 mol), followed by CuI (1.2 g, 0.006 mol), and 4-hydroxy-L-proline (1.6 g, 0.01 mol). The reaction mixture was stirred at 80°C for 14 hours. The reaction mixture was cooled to room temperature, quenched by the addition of water (100 mL), and extracted with ethyl acetate (3 x 200 mL). The organic layers were combined, washed with water (100 mL x 2), brine (100 mL), dried over sodium sulfate, and evaporated in vacuo. The crude product was purified by column chromatography using petroleum ether and ethyl acetate (7:3) as eluent to afford the title compound (3.5 g, 27%) as a brown liquid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.37-7.35 (dd, J=4.0, 5.2Hz, 1H), 7.31-7.29 (d, J=8.0Hz, 1H), 7.14-7.13 (dd, J=4.0, 5.2Hz, 1H), 5.09 (bs, 2H), 1.47 (s, 9H).

Step 7: 3-Thienylhydrazine hydrochloride

To a solution of tert-butyl 1-(3-thienyl)hydrazinecarboxylate (3.5 g, 0.0163 mol) in diethyl ether (10 mL) was added a solution of HCl in hexanes (30 mL) with stirring at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 8 hours. The organic solvent was removed under reduced pressure to yield the desired compound (2.4 g, 97%) as a light brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ10.08 (bs, 3H), 8.20 (bs, 1H), 7.48-7.46 (dd, J=3.2, 5.2Hz, 1H), 6.87-6.85 (dd, J=1.2, 4.8Hz, 1H), 6.72-6.71 (dd, J=1.6, 3.2Hz, 1H).

Step 8: Ethyl 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl (6-bromo-7-methoxy-4-oxo-2H-chromen-3(4H)-ylidene)(hydroxy)acetate (8.0 g, 0.0224 mol) in a mixture of ethanol (200 mL) and acetic acid (200 mL) was added 3-thienylhydrazine hydrochloride (4.4 g, 0.0291 mol) at room temperature under nitrogen. The reaction mixture was stirred at 100°C for 4 hours. The reaction mixture was concentrated under high vacuum. The residue was dissolved in ethyl acetate (40 mL), washed with water (20 mL), brine (20 mL), dried over sodium sulfate, and concentrated under vacuum. The crude product was purified by column chromatography using petroleum ether/ethyl acetate as eluent to afford the desired compound (8.5 g, 87%) as a light yellow solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.57-7.55 (dd, J=4.0, 5.2Hz, 1H), 7.52-7.50 (dd, J=4.0, 5.2Hz, 1H), 7.22-7.21 (dd, J=1.2, 5.2Hz, 1H), 6 .94 (s, 1H), 6.60 (s, 1H), 5.56 (s, 2H), 4.47-4.41 (dd, J=8.0, 12Hz.2H), 3.88 (s, 3H), 1.42 (t, J=8.0Hz, 3H).

Step 9: 8-Bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of ethyl 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (3 g, 0.0069 mol) in a mixture of THF (70 mL), water (20 mL), and methanol (10 mL) was added _LiOH.H₂O (0.857 g, 0.0207 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was evaporated and acidified with 1.5 N HCl solution. The solid was filtered and dried to yield the desired compound (2.8 g, 99%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.28 (bs, 1H), 8.01-8.00 (dd, J＝1.2, 5.2Hz, 1H), 7.87-7.85 (dd, J＝4.0, 5.2Hz, 1H), 7.35-7.33 (dd, J=4.0, 8.0Hz, 1H), 6.83 (s, 1H), 6.73 (s, 1H), 5.50 (s, 2H), 3.82 (s, 3H). m/z: 407[M+H] ⁺ .

Step 10: 8-Bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (4)

To a solution of 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylic acid (2.8 g, 0.0069 mol) in DCM (50 mL) was added n-tert-butylmethylamine (718 mg, 0.0083 mol), HATU (3.14 g, 0.0083 mol) and diisopropylethylamine (1.8 mL, 0.0103 mol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with sodium bicarbonate (10 mL, 10%) and extracted with DCM (2 x 50 mL). The organic layers were combined, washed with NaHCO ₃ solution (1 x 100 mL, 10% solution), brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether and ethyl acetate (9:1) as eluent to afford the desired compound (3.2 g, 98%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.98-7.97 (dd, J=1.4, 3.2Hz, 1H), 7.80-7.85 (dd, J=3.2, 4.7Hz, 1H), 7.33-7.32 (dd, J=1. 3, 5.1Hz, 1H), 6.83 (s, 1H), 6.76 (s, 1H), 5.37 (s, 2H), 3.81 (s, 3H), 3.15 (s, 3H), 1.47 (s, 9H). m/z: 476[M+H] ⁺ .

Step 11: 7-Methoxy-8-(1H-pyrazol-4-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (1)

To a solution of 8-bromo-N-(tert-butyl)-7-methoxy-N-methyl-1-(thiophen-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxamide (1 g, 0.0021 mol) in dioxane (20 mL) was added 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (611 mg, 0.0031 mol), _PdCl2 (dppf)CH2Cl2 (86 mg, 0.0001 mol), and cesium fluoride ₍ 800 _mg , 0.0053 mol) at room temperature under nitrogen. The reaction mixture was denitrogenated for 20 minutes, and water was added at room temperature. The reaction mixture was stirred at 100°C for 12 hours. The reaction mixture was filtered through celite and washed with DCM (20 mL). The filtrate was concentrated under vacuum; the crude product was dissolved in DCM (200 mL), washed with water (10 mL), brine (10 mL), and dried over sodium sulfate. The organic solvent was removed under vacuum; the crude product was purified by column chromatography using petroleum ether:ethyl acetate as eluent to afford the desired compound (0.5 g, 51%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.02-8.01 (dd, J＝1.4, 3.2Hz, 1H), 7.89-7.87 (dd, J＝3.2, 5.1Hz, 1H), 7.54 (bs, 2H), 7.37-7.36 (d d, J=1.4, 5.1Hz, 1H), 6.85 (s, 1H), 6.74 (s, 1H), 5.35 (s, 2H), 3.83 (s, 3H), 3.16 (s, 3H), 1.42 (s, 9H). m/z: 464[M+H] ⁺ .

Example 2

7-Methoxy-8-pyridin-3-yl-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (2)

To a solution of 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (Example 1, Step 10) (100 mg, 0.2 mmol) in DME (10 mL) was added pyridine-3-boronic acid (52 mg, 0.4 mmol), tetrakis(triphenylphosphine)palladium (13 mg, 0.01 mmol) and potassium carbonate (90 mg, 0.6 mmol) at room temperature and under nitrogen atmosphere. The reaction mixture was denitrogenated for 10 minutes and water (1 mL) was added. The reaction mixture was stirred at 90° C. for 16 hours. The reaction mixture was filtered through celite. The filtrate was concentrated under vacuum; the crude product was dissolved in DCM (200 mL), washed with water (10 ml), brine (10 mL), and dried over sodium sulfate. The solvent was removed under vacuum to obtain the crude product. The crude product was slurried in diethyl ether (5 mL), filtered, and dried to afford the desired compound (95 mg, 98%) as a cream solid.

¹ H NMR (400MHz, CDCl ₃ )δ8.50-8.47 (m, 2H), 7.67-7.64 (m, 1H), 7.53-7.52 (dd, J＝1.2, 3.2Hz, 1H), 7.49-7.47 (dd, J＝3.2, 5.2Hz , 1H), 7.27-7.22(m, 2H), 6.77(s, 1H), 6.69(s, 1H), 5.52(s, 2H), 3.82(s, 3H), 3.28(s, 3H), 1.52(s, 9H). m/z: 475[M+H] ⁺ .

Example 3

(4-Cyclobutanecarbonyl-[1,4]diazepan-1-yl)-(7-methoxy-8-pyridin-3-yl-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone (3)

Step 1: (8-Bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-(4-cyclobutanecarbonyl-[1,4]diazepan-1-yl)-methanone

To a solution of 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid (Example 1, Step 9) (0.5 g, 0.001 mol) in DCM (10 mL) was added 1-(cyclobutylcarbonyl)-1,4-diazepane (0.25 g, 0.001 mol), _T₃P (1 mL, 0.001 mol, 50% solution in ethyl acetate), and TEA (0.2 mL, 0.003 mol) at room temperature under nitrogen. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with saturated sodium bicarbonate solution (10 mL) and extracted with DCM (2 x 25 mL). The organic layers were combined, washed with _NaHCO₃ solution (1 x 100 mL, 100%), brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo and the crude product was purified by column chromatography using petroleum ether and ethyl acetate (9:1) as eluent to afford the desired compound as (0.4 g, 57%) as a creamy white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.51-7.48 (m, 2H), 7.22-7.20 (m, 1H), 6.99 (t, J=4.0Hz, 1H), 6.60 (t, J=2.4Hz, 1H), 5.54-5.51 (dd, J=4.0, 12.0Hz, 2H), 4.18-4.17( m, 2H), 3.86 (s, 3H), 3.81-3.79 (m, 3H), 3.71-3.60 (m, 3H), 3.57-3.54 (m, 1H), 2.36-2.34 (m, 2H), 1.99-1.95 (m, 2H), 1.94-1.90 (m, 4H). m/z: 571[M+H] ⁺ .

Step 2: (4-Cyclobutanecarbonyl-[1,4]diazepan-1-yl)-(7-methoxy-8-pyridin-3-yl-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone

To a solution of (4-cyclobutanecarbonyl-[1,4]diazepan-1-yl)-(7-methoxy-8-pyridin-3-yl-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl)-methanone (250 mg, 0.4 mmol) in DME (10 mL) was added pyridine-3-boronic acid (100 mg, 0.8 mmol), tetrakis(triphenylphosphine)palladium (30 mg, 0.02 mmol), and potassium carbonate (150 mg, 0.001 mol) at room temperature under nitrogen. The reaction mixture was denitrogenated for 10 minutes, and water (2 mL) was added. The reaction mixture was stirred at 90°C for 16 hours. The reaction mixture was filtered through celite and washed with DCM (20 mL). The filtrate was concentrated in vacuo, and the crude product was dissolved in DCM (200 mL), washed with water (10 mL), brine (10 mL), and dried over sodium sulfate. The solvent was removed in vacuo and the crude product was slurried in diethyl ether (10 mL), filtered, and dried to afford the desired compound (240 mg, 96%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ8.49-8.47 (dd, J=1.6, 4.8Hz, 2H), 7.67 (t, J=2.0Hz, 1H), 7.52-7.47 (m, 2H), 7.27-7.24 (m, 2H), 6.83-6.80 (dd, J=1.6, 4.8Hz, 1H), 6.68 (t, J=4.0, Hz, 1H), 5.59-5.56 (dd, J=1.6, 10.4Hz, 2H), 4.18-4.17(m, 2H), 3.86(s, 4H), 3.81-3.79(m, 1H), 3.71-3.60(m, 2H), 3.57- 3.54 (m, 1H), 3.36-3.34 (m, 1H), 2.36-2.349m, 2H), 2.12-1.99 (m, 6H), 0.98-0.97 (m, 1H). m/z: 570[M+H] ⁺ .

Example 4

8-Isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (5)

Step 1: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ethyl ester

To a solution of ethyl 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (Example 1, Step 8) (1 g, 0.002 mol) in THF (20 mL) was added 2,4,6-tris-(2-methyl-propenyl)-cyclotrisiloxane pyridine complex (380 mg, 0.001 mol), bis(triphenylphosphine)palladium(II) dichloride (80 mg, 0.1 mmol), and potassium triphosphate (63 mg, 0.004 mol) at room temperature under nitrogen. The reaction mixture was denitrogenated for 10 minutes, and water (2 mL) was added at room temperature. The reaction mixture was stirred at 70°C for 4 hours. The reaction mixture was filtered through celite and washed with DCM (50 mL). The filtrate was concentrated under vacuum; the crude product was dissolved in DCM (200 mL), washed with water (20 mL), brine (20 mL), and dried over sodium sulfate. The organic solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether:ethyl acetate as eluent to afford the desired product (0.9 g, 96%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.00 (m, 1H), 7.83-7.82 (d, J=3.2Hz, 1H), 7.33-7.32 (dd, J=1.6, 5.2Hz, 1H), 6.65 (s, 1H), 6.56 (s, 1H) , 6.00 (s, 1H), 5.56 (s, 2H), 4.32-4.26 (m, 2H), 3.78 (s, 3H), 1.76 (s, 3H), 4.52 (s, 3H), 1.23-1.18 (m, 3H). m/z: 411.5[M+H] ⁺ .

Step 2: 8-Isobutyl-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl 7-methoxy-8-(2-methylprop-1-en-1-yl)-1-(thiophen-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (step 1) (1 g, 0.02 mol) in a mixture of methanol and ethyl acetate (40 mL) was added palladium on carbon (10%, 0.5 g). The reaction mixture was hydrogenated at room temperature under 3 bar of hydrogen pressure for 4 hours. The reaction mixture was filtered through celite, and the filtrate was concentrated in vacuo. The residue was purified by column chromatography using petroleum ether:ethyl acetate as eluent to afford the title compound (0.7 g, 70%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.97-7.96 (dd, J＝1.2, 3.2Hz, 1H), 7.83-7.82 (dd, J＝3.2, 5.2Hz, 1H), 7.31-7.30 (dd, J＝1.2, 5.2Hz, 1H), 6.64 (s, 1H), 6.37 (s, 1H) , 5.43 (s, 2H), 4.32-4.27 (m, 2H), 3.75 (s, 3H), 2.12-2.10 (m, 2H), 1.62-1.55 (m, 1H), 1.31-1.22 (m, 3H), 0.85-0.83 (d, J=4.0Hz, 6H). m/z: 413 [M+H] ⁺ .

Step 3: 8-Isobutyl-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of ethyl 8-isobutyl-7-methoxy-1-(thiophen-3-yl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylate (250 mg, 0.006 mol) in a mixture of THF (21 mL), water (6 mL) and methanol (3 mL) was added LiOH (0.08 g, 0.001 mol) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was evaporated and acidified with 1.5N HCl solution. The solid was filtered off and dried in vacuo to give the desired compound (200 mg, 87%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.02 (bs, 1H), 7.96-7.95 (dd, J＝1.6, 3.2Hz, 1H), 7.83-7.81 (dd, J＝3.2, 5.2Hz, 1H), 7.31-7.29 (dd, J＝1.2, 5.2Hz, 1H) , 6.64 (s, 1H), 6.37 (s, 1H), 5.42 (s, 2H), 3.73 (s, 3H), 2.12-2.11 (m, 2H), 1.62-1.55 (m, 1H), 0.72-.711 (d, J=4.0Hz, 6H). m/z: 385[M+H] ⁺ .

Step 4: 8-Isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (5)

To a solution of 8-isobutyl-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid (step 3) (180 mg, 0.4 mmol) in DCM (20 mL) was added n-tert-butylmethylamine (50 mg, 0.5 mmol), HATU (0.22 g, 5 mmol) and diisopropylethylamine (0.2 mL, 0.7 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with sodium bicarbonate (10 mL, 10%) and extracted with DCM (2 x 25 mL). The organic layers were combined, washed with _NaHCO3 solution (1 x 100 mL, 10%), brine (100 mL), and dried over anhydrous sodium sulfate. The organic solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether and ethyl acetate (9:1) as eluent to afford the desired compound (140 mg, 66%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.91-7.90 (dd, J＝1.6, 3.2Hz, 1H), 7.80-7.85 (dd, J＝3.2, 5.2Hz, 1H), 7.29-7.28 (dd, J＝1.2, 5.2Hz, 1H), 6.64 (s, 1H), 6.40( s, 1H), 5.29 (s, 2H), 3.73 (s, 3H), 3.14 (s, 3H), 2.13-2.11 (m, 2H), 1.63-1.55 (m, 1H), 1.41 (s, 9H), 0.72-.711 (d, J=4.0Hz, 6H). m/z: 454[M+H] ⁺ .

Example 5

7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (6)

Step 1: 8-Bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (4)

To a solution of 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid (Example 1, Step 9) (150.00 mg; 0.37 mmol; 1.00 eq.) in DCM (0.50 ml; 7.80 mmol; 21.18 eq.) was added N,N-diisopropylamine (0.12 ml; 0.74 mmol; 2.00 eq.), O-benzotriazole-n,n,n',n'-tetramethyluronium tetrafluoroborate (TBTU) (236.53 mg; 0.74 mmol; 2.00 eq.) and n-tert-butyl-methylamine (48.16 mg, 0.55 mmol, 1.5 eq.). The reaction mixture was stirred at room temperature for 2 hours. The crude product was purified by column chromatography (Biotage) using ethyl acetate/hexane as eluent to afford the desired compound (120 mg, 68%) as a white solid.

Step 2: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (6)

To 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (50.00 mg; 0.10 mmol; 1.00 eq.) (Example 5, Step 1) in a microwave vial were added 2,4,6-tris-(2-methyl-propenyl)-cyclotrisiloxane pyridine (51.14 mg; 0.16 mmol; 1.50 eq.), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ii) complex with dichloromethane (1:1) (8.57 mg; 0.01 mmol; 0.10 eq.), dioxane (1.00 ml; 11.74 mmol; 111.82 eq.) and cesium carbonate (0.16 ml; 0.31 mmol; 3.00 eq., 3 M). The vessel was sealed, evacuated, and refluxed with nitrogen (3 times). The reaction mixture was microwaved at 120°C for 2 hours. The crude product was purified by column chromatography (Biotage) using ethyl acetate/hexane as eluent to give the desired compound (6.6 mg, 14%) as a white solid.

¹ H NMR (400MHz, MeOD) δ7.75 (dd, J=3.2, 1.4Hz, 1H), 7.69 (dd, J=5.1, 3.2Hz, 1H), 7.27 (dd, J=5.1, 1.4Hz, 1H), 6.70 (s, 1H), 6. 63 (s, 1H), 6.07 (s, 1H), 5.33 (s, 2H), 3.80 (s, 3H), 3.20 (s, 3H), 1.80 (d, J=1.3Hz, 3H), 1.54 (s, 9H), 1.50 (d, J=1.2Hz, 3H). m/z: 452[M+H] ⁺ .

Example 6

7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (2-acetylamino-ethyl)-amide (7)

Step 1: Ethyl 7-methoxy-8-(2-methylprop-1-en-1-yl)-1-(thien-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylate (6 g, 0.0138 mol) in THF (100 mL) was added 2,4,6-tris-(2-methyl-propenyl)-cyclotrisiloxane pyridine complex (5.8 g, 0.018 mol), bis[tris(4-(heptadecafluorooctyl)phenyl)phosphine]dichloropalladium(II) (1.0 g, 1.38 mmol) and potassium triphosphate (3.8 g, 0.0138 mol) at room temperature under nitrogen atmosphere. 0.0276 mol). The reaction mixture was denitrogenated for 10 minutes, and water (10 mL) was added at room temperature. The reaction mixture was heated at 80°C for 8 hours. The reaction mixture was filtered through celite and washed with DCM (50 mL). The filtrate was concentrated in vacuo; the crude product was dissolved in DCM (200 mL), washed with water (20 mL), brine (20 mL), and dried over sodium sulfate. The organic solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether:ethyl acetate as eluent to afford the desired compound (5.5 g, 98%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.00 (m, 1H), 7.83-7.82 (d, J=3.2Hz, 1H), 7.33-7.32 (dd, J=1.6, 5.2Hz, 1H), 6.65 (s, 1H), 6.56 (s, 1H) , 6.00 (s, 1H), 5.56 (s, 2H), 4.32-4.26 (m, 2H), 3.78 (s, 3H), 1.76 (s, 3H), 4.52 (s, 3H), 1.23-1.18 (m, 3H). m/z: 411.5[M+H] ⁺ .

Step 2: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of ethyl 7-methoxy-8-(2-methylprop-1-en-1-yl)-1-(thiophen-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (7 g, 0.0171 mol) in a mixture of THF (70 mL), water (20 mL), and methanol (10 mL) was added _LiOH.H₂O (2.1 g, 0.0512 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was evaporated and acidified with 1.5 N HCl solution. The solid was filtered and dried under high vacuum to give the desired compound (5.5 g, 85%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.02 (bs, 1H), 7.96-7.95 (dd, J＝1.6, 3.2Hz, 1H), 7.83-7.81 (dd, J＝3.2, 5.2Hz, 1H), 7.31-7.29 (dd, J＝1.2, 5.2Hz, 1H) , 6.64 (s, 1H), 6.37 (s, 1H), 5.42 (s, 2H), 3.73 (s, 3H), 2.12-2.11 (m, 2H), 1.62-1.55 (m, 1H), 0.72-.711 (d, J=4.0Hz, 6H). m/z: 383[M+H] ⁺ .

Step 3: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (2-acetylamino-ethyl)-amide (7)

To a solution of 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (30 mg, 0.08 mmol) in DCM (1.00 ml; 15.60 mmol; 198.87 eq.) was added N,N-diisopropylethylamine (0.02 ml; 0.09 mmol; 1.20 eq.), N-acetylethylenediamine (9.61 mg, 0.09 mmol, 1.2 eq.), and T ₃ P (0.03 ml; 0.12 mmol; 1.50 eq.). The reaction mixture was stirred at room temperature for 1.5 hours. The crude product was purified by column chromatography using ethyl acetate/hexane as the eluent to afford the desired compound (27.4 mg, 73%) as a white solid.

¹ H NMR (500MHz, CD ₃ OD) δ7.74 (dd, J=3.1, 1.2Hz, 1H), 7.67 (dd, J=5.0, 3.2Hz, 1H), 7.26 (dd, J=5.1, 1.1Hz, 1H), 6.66 (s, 1H), 6.58 (s, 1H), 6 .04(s, 1H), 5.48(s, 2H), 3.76(s, 3H), 3.46(t, J=6.0Hz, 2H), 3.41–3.33(m, 2H), 1.94(s, 3H), 1.78(s, 3H), 1.48(s, 3H). m/z: 467[M+H] ⁺ .

The following compounds were prepared according to the procedure described in Example 6.

The following compounds were prepared according to the method described in Example 4:

Process 2:

Example 7

8-Isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (19)

Step 1: 3-Chloro-1-(2-hydroxy-4,5-dimethoxyphenyl)propan-1-one

To a stirred suspension of 3,4-dimethoxyphenol (10 g, 0.06 mol) and chloropropionyl chloride (12.5 g, 0.12 mol) at 60°C under a nitrogen atmosphere was added boron trifluoride etherate (8.8 mL, 0.06 mol). After the addition was complete, the reaction mixture was heated at 70°C for 1 hour. The reaction mixture was cooled to room temperature, quenched with ice water (200 mL), and extracted with DCM (500 mL). The aqueous phase was re-extracted with 2 x 100 mL of DCM, and the organic layer was dried over sodium sulfate and concentrated in vacuo. The crude product was purified by column chromatography using petroleum ether/ethyl acetate (8:2) as the eluent to afford the desired compound (12 g, 76%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.2 (bs, 1H), 7.27 (s, 1H), 6.55 (s, 1H), 4.12-4.10 (m, 2H), 3.79 (s, 3H), 3.71 (s, 3H), 2.48-2.43 (m, 2H).

Step 2: 6,7-dimethoxy-2,3-dihydro-4H-chromen-4-one

To a solution of 3-chloro-1-(2-hydroxy-4,5-dimethoxyphenyl)propan-1-one (13 g, 0.05 mol) in ethanol was added _{dry K₂CO₃} (16.3 g, 0.10 mol) with stirring at room temperature under a nitrogen atmosphere. The resulting suspension was stirred at room temperature for 16 hours. The reaction mixture was filtered and concentrated in vacuo. The crude product was dissolved in ethyl acetate (200 mL), washed with 5% sodium bicarbonate (50 mL), brine (50 mL), and dried over sodium sulfate. The organic solvent was concentrated; the residue was purified by column chromatography using petroleum ether/ethyl acetate (8:2) as the eluent to afford the desired compound (9 g, 81%) as a light brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.12 (s, 1H), 6.59 (s, 1H), 4.47-4.44 (m, 2H), 3.80 (s, 3H), 3.72 (s, 3H), 2.69-2.65 (m, 2H).

Step 3: 6,7-dihydroxy-2,3-dihydro-4H-benzopyran-4-one

A mixture of 6,7-dimethoxy-2,3-dihydro-4H-chromen-4-one (2.5 g, 0.01 mol) and pyridine hydrochloride (20 g, 0.20 mol) was heated at 170°C for 12 hours under a nitrogen atmosphere. The reaction mixture was slurried in DCM (100 mL), the solid formed was filtered, and the filtrate was concentrated in vacuo. The crude product was purified by column chromatography using petroleum ether/ethyl acetate (5:5) as the eluent to afford the desired compound (1 g, 50%) as a pale white solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.72 (bs, 2H), 7.04 (s, 1H), 6.30 (s, 1H), 4.37-4.34 (m, 2H), 2.60-2.57 (m, 2H).

Step 4: 6-Hydroxy-7-methoxy-2,3-dihydro-4H-chromen-4-one

To a solution of 6,7-dihydroxy-2,3-dihydro-4H-chromen-4-one in DMF (60 mL) at room temperature under nitrogen was added _{dry K₂CO₃} (4.2 g, 0.0305 mol). The reaction mixture was stirred at room temperature for 15 minutes, followed by the dropwise addition of iodomethane (1.3 mL, 0.0214 mol) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered, and the filtrate was dried under vacuum. The crude product was purified by column chromatography using petroleum ether/ethyl acetate (9:1) as the eluent to afford the desired compound (4 g, 68%) as a light brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.05 (bs, 1H), 7.05 (s, 1H), 6.53 (s, 1H), 4.43-4.40 (m, 2H), 3.80 (s, 3H), 2.64-2.61 (m, 2H).

Step 5: 6-Isopropoxy-7-methoxy-2,3-dihydro-4H-chromen-4-one

To a solution of 6-hydroxy-7-methoxy-2,3-dihydro-4H-chromen-4-one (4.0 g, 0.0206 mol) in DMF (80 mL) was added dry _K₂CO₃ (5.7 g, 0.0412 mol) with stirring at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 15 minutes, followed by the dropwise addition of 2- _iodopropane (6.2 mL, 0.0618 mol) at room temperature. The reaction mixture was stirred at 65°C for 8 hours. The reaction mixture was filtered, and the filtrate was dried under vacuum. The crude product was dissolved in ethyl acetate (2 x 200 mL), washed with water (50 mL), brine (50 mL), and dried over sodium sulfate. The solvent was concentrated under vacuum, and the crude product was purified by column chromatography using petroleum ether/ethyl acetate (8:2) as the eluent to yield the desired compound (4.1 g, 87%) as a light brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.13 (s, 1H), 6.59 (s, 1H), 4.48-4.40 (m, 3H), 3.80 (s, 3H), 2.68-2.65 (m, 2H), 1.21 (s, 3H), 1.20 (s, 3H).

Step 6: Ethyl (2Z)-hydroxy(6-isopropoxy-7-methoxy-4-oxo-2H-chromen-3(4H)-ylidene)acetate

Diisopropylamine (6.9 mL, 0.0495 mol) was dissolved in dry THF (150 mL) at room temperature under a nitrogen atmosphere. The reaction mixture was cooled to -78°C, and n-butyllithium (1.6 M solution in hexane, 28.6 mL, 0.0457 mol) was added dropwise over 30 minutes. After the addition was complete, the reaction mixture was stirred at the same temperature for 15 minutes, then slowly warmed to -10°C and stirred for an additional 10 minutes. The reaction mixture was again cooled to -78°C, and a solution of 6-isopropoxy-7-methoxy-2,3-dihydro-4H-chromen-4-one (9 g, 0.0381 mol) in THF (50 mL) was added dropwise over 30 minutes, followed by stirring at -78°C. After 1 hour, diethyl oxalate (7.8 mL, 0.0571 mol) was added dropwise at -78°C. The reaction mixture was then slowly warmed to 0°C and stirred for 1 hour. The reaction mixture was cooled to -5°C, quenched with 1.5N HCl, and extracted with ethyl acetate (100 mL x 2). The organic layers were combined, washed with water (100 mL), brine (50 mL), dried over sodium sulfate, and concentrated to give the desired compound (10.5 g, 92%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ.7.17-7.13 (d, J=16Hz, 1H), 6.61-6.59 (d, J=24Hz, 1H), 5.16-5.12 (m, 1H), 4.47-4.24 (m, 1H), 4.21 (s, 3H), 2.97 (s, 3H), 2.54-5.53 (m, 1H), 1.21 (s, 6H).

Step 7: 8-Isopropoxy-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl (2Z)-hydroxy(6-isopropoxy-7-methoxy-4-oxo-2H-chromen-3(4H)-ylidene)acetate (6.0 mg, 0.0223 mol) in a mixture of ethanol (150 mL) and acetic acid (150 mL) was added 3-thienylhydrazine hydrochloride (2.7 g, 0.0223 mol) at room temperature under nitrogen. The reaction mixture was stirred at 100°C for 4 hours. The reaction mixture was concentrated under high vacuum. The residue was dissolved in ethyl acetate (20 mL), washed with water (20 mL), brine (20 mL), dried over sulfuric acid, and concentrated under vacuum. The crude product was purified by column chromatography using petroleum ether/ethyl acetate as eluent to afford the desired compound (6.5 g, 88%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.02-8.01 (dd, J＝1.2, 2.4Hz, 1H), 7.86-7.83 (dd, J＝4, 16Hz, 1H), 7.35-7.34 (dd, J＝1.2, 4Hz, 1H), 6.68 (s, 1H) , 6.19 (s, 1H), 5.40 (s, 2H), 4.31-4.26 (m, 2H), 3.94-3.88 (m, 1H), 3.72 (s, 3H), 1.31-1.287 (m, 3H), 1.07 (s, 6H).

Step 8: 8-Isopropoxy-7-methoxy-1-(3-thienyl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of 8-isopropoxy-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (4 g, 0.0108 mol) in a mixture of THF (70 mL), water (20 mL), and methanol (10 mL) was added _LiOH.H₂O (1.4 g, 0.0326 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was evaporated and acidified with 1.5 N HCl solution. The filtrate was dried under high vacuum to yield the desired compound (3.3 g, 79%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.16 (bs, 1H), 8.01-8.00 (dd, J=1.2, 2.4Hz, 1H), 7.85-7.83 (dd, J=4, 16Hz, 1H), 7.35-7.33 (dd, J =1.2, 4Hz, 1H), 6.69 (s, 1H), 6.19 (s, 1H), 5.39 (s, 2H), 3.94-3.88 (m, 2H), 3.72 (s, 3H), 1.07 (s, 6H).

Step 9: 8-Isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (19)

To a solution of 8-isopropoxy-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid (1.1 g, 0.0028 mol) in DCM (50 mL) was added N-tert-butylmethylamine (0.3 g, 0.0034 mol), HATU (1.3 g, 0.0034 mol), and DIPEA (0.8 mL, 0.0043 mol) with stirring at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (2 x 100 mL). The organic layers were combined, washed with brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether/ethyl acetate (8:2) as the eluent to afford the desired compound (1.1 g, 85%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.97-7.96 (dd, J＝1.2, 2.4Hz, 1H), 7.82-7.80 (dd, J＝4, 16Hz, 1H), 7.34-7.32 (dd, J＝1.2, 4Hz, 1H), 6.69 (s, 1H), 6.22 (s, 1H), 5.27 (s, 2H), 3.39-3.89 (m, 1H), 3.72 (s, 3H), 3.14 (s, 3H), 1.41 (s, 9H), 1.06 (s, 6H).

m/z: 456 [M+H] ⁺ .

The following compounds were prepared according to the procedure described in Example 7.

The following compounds were prepared according to the method described in Example 1.

Example 8

7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid methyl-[2-(2-oxo-oxazolidin-3-yl)-ethyl]-amide (23)

To 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid [2-(2-oxo-oxazolidin-3-yl)-ethyl]-amide (22.00 mg; 0.04 mmol; 1.00 eq.) was added N,N-dimethylformamide (0.50 ml) and sodium hydride (2.67 mg; 0.07 mmol; 1.50 eq.). The reaction mixture was stirred at room temperature for 10 minutes, then iodomethane (0.00 ml; 0.05 mmol; 1.10 eq.) (1 drop) was added, and the reaction mixture was stirred at room temperature for 15 minutes. Water was added, and the product was extracted with ethyl acetate and washed with water. The organic layer was dried, filtered, and concentrated. The residue was dried under high vacuum and then lyophilized to yield the desired product (22.5 mg, 99%) as a creamy white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.56–7.42(m, 2H), 7.24(d, J＝4.9Hz, 1H), 6.73(d, J＝4.0Hz, 1H), 6.57(d , J=2.8Hz, 1H), 6.12 (s, 1H), 5.55 (s, 1H), 5.51 (s, 1H), 4.38–4.31 (m, 1H), 4 .20–4.10(m, 2H), 3.82(s, 3H), 3.81–3.72(m, 2H), 3.62–3.55(m, 3H), 3.53( s, 1H), 3.46–3.38 (m, 1H), 3.17 (s, 1H), 1.85 (s, 3H), 1.53 (d, J=3.8Hz, 3H). m/z: 509[M+H] ⁺ .

The following compounds were prepared according to the method disclosed in Example 8:

Example 9

(8-Isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-[4-(tetrahydro-furan-2-carbonyl)-[1,4]diazepan-1-yl]-methanone (21)

Step 1: To a solution of 8-isobutyl-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid (70.00 mg; 0.18 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 85.68 eq.) was added N,N-diisopropylethylamine (0.04 ml; 0.22 mmol; 1.20 eq.), 1-boc-hexahydro-1,4-diazepane (0.04 ml; 0.22 mmol; 1.20 eq.), and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.08 ml; 0.27 mmol; 1.50 eq.). After stirring at room temperature for 2 hours, the reaction mixture was washed with water and 1N HCl. The organic layer was dried over sodium sulfate, filtered, and concentrated to afford tert-butyl 4-(8-isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carbonyl)-[1,4]diazepane-1-carboxylate (103.00 mg; 0.18 mmol) as a white foam. To a solution of tert-butyl 4-(8-isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carbonyl)-[1,4]diazepane-1-carboxylate (103.00 mg; 0.18 mmol) in methanol (1.00 ml; 24.69 mmol; 135.58 eq.) was added hydrochloric acid (0.46 ml; 1.82 mmol; 10.00 eq.) (4M in dioxane). After stirring for 4 hours, the reaction mixture was concentrated to dryness and diluted with water. The mixture was lyophilized to give [1,4]diazepan-1-yl-(8-isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone as a white solid.

Step 2: To a solution of [1,4]-diazepan-1-yl-(8-isobutyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl)-methanone (30.00 mg; 0.06 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 242.64 eq.) were added N,N-diisopropylethylamine (0.01 ml; 0.08 mmol; 1.20 eq.), tetrahydro-2-furoic acid (0.01 ml; 0.13 mmol; 2.00 eq.) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.03 ml; 0.10 mmol; 1.50 eq.). After stirring at room temperature for 30 minutes, the reaction was concentrated to dryness and purified by flash chromatography to give the desired compound (10.7 mg, 30%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.51–7.44(m, 2H), 7.50–7.44(m, 1H), 7.25–7.21(m, 1H), 6.59(d, J＝7.0Hz, 1H), 6.56(d, J＝2 .7Hz, 1H), 5.50 (dd, J=4.9, 3.5Hz, 2H), 4.70–4.58 (m, 1H), 4.53–4.41 (m, 1H), 4.41–4.22 (m, 1H) , 4.19–4.09(m, 1H), 4.08–3.82(m, 4H), 3.79(s, 3H), 3.71–3.59(m, 2H), 3.58–3.42(m, 1H), 2.3 6–2.26 (m, 1H), 2.26–2.20 (m, 2H), 2.19–1.83 (m, 3H), 1.78–1.60 (m, 2H), 0.81 (d, J=6.6Hz, 6H). m/z: 565[M+H] ⁺ .

Example 10

7-Methoxy-8-(1H-pyrazol-4-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid methylamide (22)

Step 1: To 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (100.00 mg; 0.25 mmol; 1.00 eq.) in a microwave vial was added tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-pyrazole-1-carboxylate (108.35 mg; 0.37 mmol; 1.50 eq.), 1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(ii) complex with dichloromethane (1:1) (20.05 mg; 0.02 mmol; 0.10 eq.) and cesium carbonate (0.25 ml; 0.49 mmol; 2.00 eq.) (3 M aqueous solution). The vessel was sealed, evacuated, and refluxed with nitrogen (3 times). The reaction mixture was heated in a microwave at 100°C for 30 minutes. The mixture was diluted with ethyl acetate and washed with water and 1N HCl. The organic layer was dried over sodium sulfate, filtered, and concentrated to afford 8-(1-tert-butoxycarbonyl-1H-pyrazol-4-yl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid as a crude product.

Step 2: To a solution of 8-(1-tert-butoxycarbonyl-1H-pyrazol-4-yl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (121.00 mg; 0.24 mmol; 1.00 eq.) in DCM (1.50 ml; 23.40 mmol; 95.64 eq.) were added DIPEA (0.09 ml; 0.49 mmol; 2.00 eq.), O-benzotriazole-n,n,n',n'-tetramethyluronium tetrafluoroborate (157.13 mg; 0.49 mmol; 2.00 eq.) and n-tert-butylmethylamine (0.06 ml; 0.49 mmol; 2.00 eq.). After stirring at room temperature for 30 minutes, the mixture was concentrated to dryness and purified by flash chromatography to give 4-[3-(tert-butyl-methyl-carbamoyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-8-yl]-pyrazole-1-carboxylic acid tert-butyl ester as a white solid (99 mg, 72%).

Step 3: To a solution of 4-[3-(tert-butyl-methyl-carbamoyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-8-yl]-pyrazole-1-carboxylic acid tert-butyl ester (99.00 mg; 0.18 mmol) in methanol (3.00 ml; 74.06 mmol; 302.68 eq.) was added hydrochloric acid (0.20 ml; 0.80 mmol; 3.27 eq.) (4M% solution in dioxane). After stirring at room temperature for 1 hour, the mixture was concentrated and purified by flash chromatography to give the desired product (33.8 mg, 34%) as a white solid.

^1H NMR (400MHz, DMSO) δ12.83 (s, 1H), 8.33 (dd, J=9.4, 4.6Hz, 1H), 8.07 (dd, J=3.2, 1.4Hz, 1H), 7.92 (dd, J=5.1, 3.2Hz, 1H), 7.77 (s, 1H), 7.41 (dd, J=5.1, 1.4Hz, 1H), 7.32 (s, 1H), 6.85 (s, 1H), 6.76 (s, 1H), 5.52 (s, 2H), 3.86 (s, 3H), 2.75 (d, J=4.7Hz, 3H). m/z: 408[M+H] ⁺ .

Example 11

[4-(Azetidine-1-carbonyl)-azepan-1-yl]-(8-isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone (26)

To a solution of 1-(8-isopropoxy-7-methoxy-1-(thiophen-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carbonyl)azepane-4-carboxylic acid (45.00 mg; 0.09 mmol; 1.00 eq.) in DCM (2.00 ml; 31.20 mmol; 354.72 eq.) were added DIPEA (0.02 ml; 0.13 mmol; 1.50 eq.), azetidine (0.01 ml; 0.13 mmol; 1.50 eq.) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.08 ml; 0.13 mmol; 1.50 eq.). After stirring at room temperature for 30 minutes, the reaction mixture was concentrated to dryness and purified by flash chromatography to give the desired product (24.5 mg, 51%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.54–7.45 (m, 2H), 7.24 (t, J=5.0Hz, 1H), 6.61 (s, 1H), 6.43 (d, J=5.8Hz, 1H), 5.55–5.41 (m, 2H), 4.59 (d, J=14.3Hz, 1H), 4.23–3.86 (m, 4 H), 3.83 (s, 3H), 3.77–3.49 (m, 1H), 3.35–3.27 (m, 1H), 2.42–2.07 (m, 4H), 2.03–1.68 (m, 6H), 1.27 (t, J=7.1Hz, 1H), 1.21 (d, J=6.1Hz, 6H). m/z: 551[M+H] ⁺ .

Example 12

8-Isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-amino-propyl)-amide (27)

Step 1: To a solution of 8-isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (120.00 mg; 0.31 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 50.24 eq.) were added N,N-diisopropylethylamine (0.07 ml; 0.37 mmol; 1.20 eq.), n-boc-1,3-diaminopropane (81.16 mg; 0.47 mmol; 1.50 eq.) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.14 ml; 0.47 mmol; 1.50 eq.). After stirring at room temperature for 30 minutes, the reaction mixture was concentrated to dryness and purified by flash chromatography to give {3-[(8-isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl)-amino]-propyl}-carbamic acid tert-butyl ester (170 mg, 100%) as a white solid.

Step 2: To a solution of {3-[(8-isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carbonyl)-amino]-propyl}-carbamic acid tert-butyl ester (172.70 mg; 0.32 mmol) in methanol (4.00 ml; 98.75 mmol; 317.98 eq.) was added hydrochloric acid in dioxane (0.31 ml; 1.24 mmol; 4.00 eq.). The mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated to dryness and purified by flash chromatography to give the desired product (71.3 mg, 52%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ8.77–8.64(m, 2H), 7.55(d, J=7.2Hz, 2H), 6.62(s, 1H), 6.36(s, 1H), 5.51(s, 2H), 4.13–3.94(m, 2H), 3.85 (s, 3H), 3.65–3.59 (m, 2H), 3.23–3.07 (m, 2H), 2.21–2.09 (m, 1H), 1.21 (d, J=6.0Hz, 6H). m/z: 443[M+H] ⁺ .

Example 13

8-Isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid [3-(cyclobutanecarbonyl-amino)-propyl]-amide (28)

To a solution of 8-isopropoxy-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (3-amino-propyl)-amide (27) (65.00 mg; 0.15 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 106.21 eq.) were added N,N-diisopropylethylamine (0.03 ml; 0.18 mmol; 1.20 eq.), cyclobutanecarboxylic acid (0.14 ml; 1.50 mmol; 10.21 eq.) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.13 ml; 0.22 mmol; 1.50 eq.). After stirring at room temperature for 18 h, the reaction mixture was concentrated to dryness and purified by flash chromatography to give the desired product (55.8 mg, 72%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.57–7.46(m, 2H), 7.23(d, J＝5.0Hz, 1H), 7.16(t, J＝6.3Hz, 1H), 6.59(s, 1H) , 6.35(m, 2H), 5.54(s, 2H), 4.02(dt, J=12.1, 6.0Hz, 1H), 3.82(s, 3H), 3.46(q, J＝6.2Hz, 1H), 3.31(q, J＝6.0Hz, 2H), 3.03(p, J＝8.6Hz, 1H), 2.36–2.22(m, 2H), 2.20–2.10 (m, 2H), 2.01–1.80 (m, 2H), 1.79–1.67 (m, 2H), 1.19 (d, J=6.1Hz, 6H). m/z: 525[M+H] ⁺ .

Example 14

Cyclobutanecarboxylic acid {1-[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-azepan-4-yl}-amide (37)

To a solution of (4-amino-azepan-1-yl)-[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl]-methanone (55.00 mg; 0.11 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 135.76 eq.) were added N,N-diisopropylethylamine (0.02 ml; 0.14 mmol; 1.20 eq.), cyclobutanecarboxylic acid (0.14 ml; 1.50 mmol; 13.05 eq.) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.10 ml; 0.17 mmol; 1.50 eq.). After stirring at room temperature for 30 minutes, the reaction mixture was washed with NaHCO ₃ and extracted with DCM. The organic layer was concentrated and purified by flash chromatography to give the desired product (60.3 mg, 94%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.53–7.44(m, 2H), 7.23(dd, J=5.0, 1.3Hz, 1H), 6.76(s, 1H), 6.57(s, 1H), 6.12(s, 1 H), 5.60–5.45(m, 3H), 4.61–4.49(m, 1H), 4.43–4.30(m, 1H), 4.18–3.99(m, 2H), 3.82( s, 3H), 3.60–3.48 (m, 1H), 3.45–3.34 (m, 1H), 3.26 (t, J=10.3Hz, 1H), 3.00–2.81 (m, 1H ), 2.31–2.06(m, 4H), 2.01–1.87(m, 2H), 1.85(s, 3H), 1.72–1.59(m, 4H), 1.54(s, 3H). m/z: 547[M+H] ⁺ .

Example 15

8-(1,2-Dihydroxy-2-methyl-propyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (38)

To a solution of 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (30.00 mg; 0.07 mmol; 1.00 eq.) in water (0.10 ml) and acetone (0.40 ml) was added 4-methylmorpholine 4-oxide (23.35 mg; 0.20 mmol; 3.00 eq.) and osmium tetroxide (0.51 mg; 0.00 mmol; 0.03 eq.). After stirring at room temperature for 1 hour, the mixture was quenched with saturated sodium sulfide solution and extracted with ethyl acetate. The organic layer was dried over sodium sulfate, filtered, concentrated, and purified by flash chromatography to yield the desired compound (22.5 mg, 70%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.51 (dd, J＝3.2, 1.3Hz, 1H), 7.47 (dd, J＝5.1, 3.2Hz, 1H), 7.21 (dd, J＝5.1, 1.3Hz, 1H), 6.90 (s, 1H), 6.56 (s, 1H), 5.53–5.38( m, 2H), 4.69 (s, 1H), 3.81 (s, 3H), 3.27 (s, 3H), 2.67 (s, 1H), 1.52 (s, 9H), 1.28 (dd, J=6.0, 3.0Hz, 1H), 1.15 (s, 3H), 0.97 (s, 3H). m/z：486[M+H] ⁺

Example 16

1-[7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxylic acid dimethylamide (41)

To a solution of 1-[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxylic acid (100.00 mg; 0.20 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 79.19 eq.) were added N,N-diisopropylethylamine (0.04 ml; 0.24 mmol; 1.20 eq.), dimethylamine hydrochloride (24.10 mg; 0.30 mmol; 1.50 eq.) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.17 ml; 0.30 mmol; 1.50 eq.). After stirring at room temperature for 30 minutes, the reaction mixture was concentrated and purified by flash chromatography to give the desired product (92.3 mg, 88%) as a white solid.

¹ H NMR (500MHz, cdcl ₃ )δ7.46–7.34 (m, 2H), 7.14 (ddd, J=13.8, 4.9, 1.4Hz, 1H), 6.67 (d, J=12.0Hz, 1H), 6.49 (d, J=1.8Hz, 1H), 6. 03 (s, 1H), 5.52–5.41 (m, 2H), 4.61–4.55 (m, 1H), 4.22–4.10 (m, 1H), 3.98–3.85 (m, 1H), 3.73 (s, 3H), 3.55– 3.48 (m, 1H), 3.45–3.37 (m, 1H), 3.22–3.14 (m, 1H), 2.96 (s, 1H), 2.88 (d, J=8.1Hz, 3H), 2.83 (s, 1H), 2.71– 2.60 (m, 1H), 2.14–2.04 (m, 1H), 1.95–1.83 (m, 2H), 1.76 (s, 3H), 1.70–1.53 (m, 2H), 1.45 (d, J=4.8Hz, 3H). m/z: 535[M+H] ⁺ .

Example 17

1-[7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxylic acid methylamide (42)

1-[7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxylic acid methylamide was prepared from a solution of 1-[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxylic acid (100.00 mg; 0.20 mmol; 1.00 eq.) in DCM (1.00 ml; 15.60 mmol; 79.19 eq.) and methylamine hydrochloride (13.30 mg; 0.20 mmol; 1.00 eq.) according to the same procedure as in Example 16. The desired compound was obtained in a 52% yield (53.5 mg) as a white solid.

¹ H NMR (500MHz, cdcl ₃ )δ7.49–7.43(m, 2H), 7.21(d, J＝5.1Hz, 1H), 6.74(s, 1H), 6.56(s, 1H), 6.10(s, 1H), 5.55(t, J＝4.9Hz, 1H), 5.52 (t, J＝4.1Hz, 2H), 4.49–4.43 (m, 1H), 4.18 (dt, J＝14.5, 5.4Hz, 1H), 4.07–4.00 (m, 1H), 3.95 (dt, J＝9.5 , 4.4Hz, 1H), 3.85–3.82 (m, 1H), 3.80 (s, 3H), 3.72–3.63 (m, 1H), 3.49–3.42 (m, 1H), 2.78 (dd, J=14.0, 4.8H z, 3H), 2.34–2.12 (m, 1H), 2.05–1.91 (m, 1H), 1.84 (d, J=1.2Hz, 3H), 1.80–1.68 (m, 2H), 1.54–1.51 (s, 3H). m/z: 521[M+H] ⁺ .

Example 18

1-[7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxamide (43)

To a solution of 1-[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-azepane-4-carboxylic acid (75.00 mg; 0.15 mmol; 1.00 eq.) in DMSO (1.00 ml) was added 1,1'-carbonylbis(1H-imidazole) (119.72 mg; 0.74 mmol; 5.00 eq.). The reaction mixture was stirred at room temperature for 18 hours, then ammonium acetate (34.17 mg; 0.44 mmol; 3.00 eq.) was added and the reaction was continued with stirring at room temperature for 2 hours. After the reaction was complete, water was added to the mixture, and the precipitated solid was filtered and washed with water to yield the desired compound (62.5 mg, 84%) as a white solid.

¹ H NMR (500MHz, cdcl ₃ )δ7.44–7.34(m, 2H), 7.15–7.11(m, 1H), 6.67(s, 1H), 6.49(s, 1H), 6.03(s, 1H), 5.47–5.43(m, 2H), 5.13 (s, 1H), 4.75 (s, 1H), 4.44–4.36 (m, 1H), 4.12 (dt, J=14.7, 5.3Hz, 1H), 4.03–3.95 (m, 1 H), 3.88 (d, J=14.3Hz, 1H), 3.83–3.74 (m, 1H), 3.73 (s, 3H), 3.66–3.56 (m, 1H), 3.43–3.37 (m, 1H ), 2.36–2.25 (m, 1H), 2.23–2.11 (m, 1H), 2.03–1.87 (m, 2H), 1.76 (d, J=1.0Hz, 3H), 1.45 (s, 3H). m/z: 507[M+H] ⁺ .

Example 19

7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid [3-(cyclobutanecarbonyl-amino)-propyl]-amide (57)

Step 1: Following the same procedure of Step 1 of Example 12, (3-{[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-amino}-propyl)-carbamic acid tert-butyl ester was prepared from 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (150 mg, 0.39 mmol) and N-boc-1,3-diaminopropane (102.5 mg, 0.59 mmol, 1.5 eq.).

Step 2: Following the same procedure of Step 2 of Example 12, 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carbonyl]-amino}-propyl)-carbamic acid tert-butyl ester and HCl (4M solution in dioxane) was prepared.

Step 3: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid [3-(cyclobutanecarbonyl-amino)-propyl]-amide was prepared from 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-amino-propyl)-amide (100 mg, 0.23 mmol) and cyclobutanecarboxylic acid (0.14 ml; 1.50 mmol; 6.58 eq.) according to the same procedure as in Step 1 of Example 12. The desired compound was obtained in 97% yield (115 mg) as a white solid.

¹ H NMR (400MHz, CDCl ₃ )δ7.55(s, 1H), 7.53–7.48(m, 1H), 7.25(d, J＝4.7Hz, 1H), 7.11(t, J＝7.1Hz, 1H), 6.71(s, 1H), 6 .58(s, 1H), 6.32–6.25(m, 1H), 6.12(s, 1H), 5.62(s, 2H), 3.83(s, 3H), 3.49(dd, J＝12.3,5.9Hz , 2H), 3.34(dd, J＝12.1, 6.0Hz, 2H), 3.12–3.00(m, 1H), 2.32(dt, J＝19.0, 9.7Hz, 2H), 2.19(dd, J=18.8, 9.9Hz, 2H), 1.98 (dd, J=18.7, 8.5Hz, 2H), 1.85 (s, 3H), 1.79–1.71 (m, 2H), 1.52 (s, 3H). m/z: 521[M+H] ⁺ .

Process 3:

Process 4:

Example 20

7-Methoxy-8-(2-methyl-propenyl)-1-thiazol-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (89)

Step 1: 2-Hydrazino-1,3-thiazole hydrochloride

To a mixture of 2-aminothiazole (10.0 g, 100 mmol) and concentrated hydrochloric acid (80 mL) was added dropwise a solution of sodium nitrite (6.90 g, 100 mmol) in water (50 mL) at -10 ° C. The reaction mixture was stirred at the same temperature for 10 minutes, and then a solution of tin chloride (II) (37.9 g, 200 mmol) in concentrated hydrochloric acid (20 mL) was carefully added dropwise so that the temperature of the solution did not exceed -10 ° C. After the addition was complete, the reaction mixture was stirred at the same temperature for 30 minutes. The crystals obtained were collected by filtration. The crystals were recrystallized in diethyl ether to obtain the desired compound (11.0 g, 97%) as a brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.74 (bs, 1H), 7.27-7.26 (d, J=4.0Hz, 1H), 7.00-6.99 (d, J=4.0Hz, 1H), 3.45 (bs, 3H). m/z: 116[M+H] ⁺ .

Step 2: Ethyl 8-bromo-7-methoxy-1-(1,3-thiazol-2-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl (6-bromo-7-methoxy-4-oxo-2H-chromen-3(4H)-ylidene)(hydroxy)acetate (3.0 g, 0.0084 mol) in a mixture of ethanol (100 mL) and acetic acid (100 mL) was added 2-hydrazino-1,3-thiazole hydrochloride (1.9 g, 0.0126 mol) at room temperature under nitrogen. The reaction mixture was stirred at 100°C for 4 hours. The reaction mixture was concentrated under high vacuum. The residue was dissolved in ethyl acetate (40 mL), washed with water (20 mL), brine (20 mL), dried over sodium sulfate, and concentrated under vacuum. The crude product was purified by column chromatography using petroleum ether/ethyl acetate as eluent to give the desired compound (1.5 g, 41%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.61 (s, 1H), 7.86-7.85 (d, J=1.4Hz, 2H), 6.86 (s, 1H), 5.45 (s, 2H), 4.3 7-4.32 (dd, J=7.0, 14.2, 2H), 3.86 (s, 3H), 1.34-1.31 (t, J=7.1, 14.2, 3H). m/z: 438[M+H] ⁺ .

Step 3: Ethyl 7-methoxy-8-(2-methylprop-1-en-1-yl)-1-(1,3-thiazol-2-yl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylate

To a solution of 8-bromo-7-methoxy-1-(1,3-thiazole-2yl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylic acid ethyl ester (2.6 g, 0.0059 mol) in THF (100 mL) was added 2,4,6-tris-(2-methyl-propenyl)-cyclotrisiloxane pyridine complex (2.9 g, 0.0089 mol), bis(triphenylphosphine)palladium(II) dichloride (417 mg, 0.0006 mol) and potassium triphosphate (2.0 g, 0.0149 mol) at room temperature and under a nitrogen atmosphere. The reaction mixture was denitrogenated for 10 minutes and water (10 mL) was added at room temperature. The reaction mixture was stirred at 90 ° C for 12 hours. The reaction mixture was filtered through celite and washed with DCM (50 mL). The filtrate was concentrated in vacuo; the crude product was dissolved in DCM (200 mL), washed with water (20 mL), brine (20 mL), and dried over sodium sulfate. The organic solvent was removed in vacuo, and the crude product was purified by column chromatography on silica gel (60-120 mesh) using (60-120) petroleum ether:ethyl acetate as eluent to afford the desired compound (2.2 g, 90%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.94 (s, 1H), 7.87-7.86 (d, J=3.5Hz, 1H), 7.82-7.81 (d, J=3.5Hz, 1H), 6.69 (s, 1H), 6.01 (s, 1H), 5.42 (s, 2H), 4.35-4.33 (d, J=7.1Hz, 2H), 3.77 (s, 3H), 1.82 (s, 3H), 1.70 (s, 3H), 1.34-1.31 (t, J=7.1, 14.2Hz, 3H). m/z: 412[M+H] ⁺ .

Step 4: 7-Methoxy-8-(2-methylprop-1-en-1-yl)-1-(1,3-thiazol-2-yl)-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of ethyl 7-methoxy-8-(2-methylprop-1-en-1-yl)-1-(1,3-thiazol-2-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (1 g, 0.0024 mol) in a mixture of THF (35 mL), water (10 mL), and methanol (5 mL) was added _LiOH.H₂O (302 mg, 0.0073 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was evaporated and acidified with 1.5 N HCl solution. The precipitated solid was filtered and dried under high vacuum to give the desired compound (900 mg, 97%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.57 (bs, 1H), 8.00 (s, 1H), 7.85-7.84 (d, J = 3.5Hz, 1H), 7.80 (d, J = 3.5Hz, 1H ), 6.69(s, 1H), 6.11(s, 1H), 5.41(s, 2H), 3.77(s, 3H), 1.83(s, 3H), 1.71(s, 3H). m/z: 384[M+H] ⁺ .

Step 5: 7-Methoxy-8-(2-methyl-propenyl)-1-thiazol-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide

To a solution of 7-methoxy-8-(2-methylprop-1-en-1-yl)-1-(1,3-thiazol-2-yl)-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (900 mg, 0.0023 mol) in DCM (50 mL) was added N-tert-butylmethylamine (225 mg, 0.0028 mol), HATU (1.1 g, 0.0028 mol) and diisopropylethylamine (0.6 mL, 0.0035 mol) at room temperature and under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with sodium bicarbonate (10 mL, 10%) and extracted with DCM (2 x 50 mL). The organic layers were combined, washed with NaHCO ₃ solution (1 x 100 mL, 10% aqueous solution), brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether and ethyl acetate (9:1) as eluent to afford the desired compound (850 mg, 80%) as a cream solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.14 (s, 1H), 7.79-7.76 (dd, J=3.5, 6.3Hz, 2H), 6.70 (s, 1H), 6.12 (s, 1H), 5 .25(s, 2H), 3.77(s, 3H), 3.12(s, 3H), 1.84(s, 3H), 1.74(s, 3H), 1.44(s, 9H). m/z: 453[M+H] ⁺ .

Example 21

7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (90)

Step 1: tert-Butyl 1-(2-thienyl)hydrazinecarboxylate

To a solution of 2-bromothiophene (10 g, 0.0613 mol) in DMSO (200 mL) at room temperature under nitrogen was added tert-butyl carbazate (16.3 g, 0.1227 mol), cesium carbonate (40 g, 0.1227 mol), followed by CuI (1.2 g, 0.0061 mol) and 4-hydroxy-L-proline (1.6 g, 0.0123 mol). The reaction mixture was stirred at 80°C for 14 hours. The reaction mixture was cooled to room temperature, quenched with water (100 mL), and extracted with ethyl acetate (3 x 200 mL). The organic layers were combined, washed with water (100 mL x 2), brine (100 mL), dried over sodium sulfate, and evaporated in vacuo. The crude product was purified by column chromatography using petroleum ether and ethyl acetate (7:3) as the eluent to afford the desired compound (3.0 g, 40%) as a brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 6.89-6.87 (dd, J=1.7, 5.4Hz, 1H), 6.81-6.79 (dd, J=3.4, 7.2Hz, 2H), 5.38 (s, 2H), 1.49 (s, 9H). m/z: 115[M+H] ⁺ .

Step 2: 2-Thienylhydrazine hydrochloride

To a solution of tert-butyl 1-(2-thienyl)hydrazinecarboxylate (4.3 g, 0.0201 mol) in dichloromethane (20 mL) was added a solution of HCl in dioxane (30 mL) with stirring at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 8 hours. The organic solvent was removed under reduced pressure to yield the desired compound (2.9 g, 96%) as a light brown solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ10.17 (bs, 3H), 8.41 (bs, 1H), 7.07-7.05 (dd, J=1.4, 5.4Hz, 1H), 6.85-6.83 (dd, J=3.6, 5.4Hz, 1H), 6.72-6.71 (dd, J=1.4, 3.7Hz, 1H). m/z: 115[M+H] ⁺ .

Step 3: Ethyl 8-bromo-7-methoxy-1-(2-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl (6-bromo-7-methoxy-4-oxo-2H-chromen-3(4H)-ylidene)(hydroxy)acetate (6.0 g, 0.0168 mol) in a mixture of ethanol (100 mL) and acetic acid (100 mL) was added 2-thienylhydrazine hydrochloride (2.9 g, 0.0252 mol) at room temperature under nitrogen. The reaction mixture was stirred at 100° C. for 4 hours. The reaction mixture was concentrated under high vacuum. The residue was dissolved in ethyl acetate (40 mL), washed with water (20 mL), brine (20 mL), dried over sodium sulfate, and dried under vacuum. The crude product was purified by column chromatography using petroleum ether/ethyl acetate as eluent to afford the desired compound (4.0 g, 55%) as a light yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.84-7.82 (dd, J=1.2, 5.5Hz, 1H), 7.49-7.47 (dd, J=1.2, 3.6Hz, 1H), 7.24-7.22 (dd, J=3.8, 5.4Hz, 1H), 6.83 (s, 1H), 6.72 (s, 1H), 5.51 (s, 2H), 4.33-4.28 (dd, J=7.1, 14.2Hz.2H), 3.82 (s, 3H), 1.32-1.28 (t, J=7.1, 14.2Hz, 3H). m/z: 437[M+H] ⁺ .

Step 4: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ethyl ester

To a solution of ethyl 8-bromo-7-methoxy-1-(2-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (3.8 g, 0.0087 mol) in THF (100 mL) at room temperature under nitrogen was added 2,4,6-tris-(2-methyl-propenyl)-cyclotrisiloxane pyridine complex (4.3 g, 0.0131 mol), (306 mg, 0.0004 mol), and potassium triphosphate (2.4 g, 0.0174 mol). The reaction mixture was denitrogenated for 10 minutes, and water (10 mL) was added at room temperature. The reaction mixture was stirred at 90°C for 12 hours. The reaction mixture was filtered through celite and washed with DCM (50 mL). The filtrate was concentrated under vacuum; the crude product was dissolved in DCM (200 mL), washed with water (20 mL), brine (20 mL), and dried over sodium sulfate. The organic solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether:ethyl acetate as eluent to afford the desired product (2.5 g, 70%) as a light yellow solid.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.79-7.77 (dd, J=1.4, 5.5Hz, 1H), 7.46-7.45 (dd, J=1.5, 3.7Hz, 1H), 7.20-7.18 (dd, J=3.7, 5.6Hz, 1H), 6.66 (s, 1H), 6.56 (s, 1H), 6.00 (s, 1H ), 5.47 (s, 2H), 4.33-4.27 (dd, J=7.1, 14.2Hz, 2H), 3.74 (s, 3H), 1.73 (d, J=1.1Hz, 3H), 1.39 (d, J=1.2Hz, 3H), 1.32-1.28 (t, J=7.1, 14.2Hz, 3H). m/z: 411[M+H] ⁺ .

Step 5: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of ethyl 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylate (1 g, 0.0024 mol) in a mixture of THF (35 mL), water (10 mL), and methanol (5 mL) was added _LiOH.H₂O (303 mg, 0.0073 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was evaporated and acidified with 1.5 N HCl solution. The precipitated solid was filtered and dried to give the desired compound (800 mg, 75%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.30 (bs, 1H), 7.78-7.76 (dd, J＝1.4, 5.6Hz, 1H), 7.45-7.44 (dd, J＝1.4, 3.7Hz, 1H), 7.19-7.17 (dd, J＝3 .7, 5.5Hz, 1H), 6.66(s, 1H), 6.56(s, 1H), 6.00(s, 1H), 5.46(s, 2H), 3.73(s, 3H), 1.73(s, 3H), 1.39(s, 3H). m/z: 383[M+H] ⁺ .

Step 6: 7-Methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide

To a solution of 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (800 mg, 0.0021 mol) in DCM (50 mL) was added N-tert-butylmethylamine (220 mg, 0.0025 mol), HATU (950 mg, 0.0025 mol) and diisopropylethylamine (0.6 mL, 0.0032 mol) at room temperature and under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with sodium bicarbonate (10 mL, 10%) and extracted with DCM (2 x 50 mL). The organic layers were combined, washed with NaHCO ₃ solution (1 x 100 mL, 10% aqueous solution), brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether and ethyl acetate (9:1) as eluent to afford the desired compound (850 mg, 90%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.75-7.73 (dd, J＝1.4, 5.5Hz, 1H), 7.42-7.41 (dd, J＝1.4, 3.6Hz, 1H), 7.18-7.15 (dd, J＝3.8, 5.6Hz, 1H) , 6.66 (s, 1H), 6.59 (s, 1H), 6.01 (s, 1H), 5.33 (s, 2H), 3.73 (s, 3H), 3.12 (s, 3H), 1.73 (s, 3H), 1.41 (s, 12H). m/z: 452[M+H] ⁺ .

Example 22

8-Isobutyl-7-methoxy-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (91)

Step 1: 8-Isobutyl-7-methoxy-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ethyl ester

To a solution of ethyl 7-methoxy-8-(2-methyl-propenyl)-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylate (1.2 g, 0.0027 mol) in a mixture of methanol and ethyl acetate (100 mL) was added palladium on carbon (20%, 0.24 g). The reaction mixture was hydrogenated at room temperature under 3 bar of hydrogen pressure for 8 hours. The reaction mixture was filtered through celite to remove the catalyst, and the filtrate was concentrated in vacuo. The residue was purified by column chromatography using petroleum ether:ethyl acetate as the eluent to afford the title compound (1.1 g, 90%) as a creamy white solid.

¹ H NMR (400 MHz, DMSO-d ₆ )δ7.81-7.79 (dd, J＝1.4, 5.5Hz, 1H), 7.43-7.41 (dd, J＝1.4, 3.7Hz, 1H), 7.20 -7.18(dd, J=3.7, 5.5Hz, 1H), 6.65(s, 1H), 6.36(s, 1H), 5.44(s, 2H), 4.33-4 .27 (dd, J=7.0, 14.2Hz, 2H), 3.74 (s, 3H), 2.12-2.10 (d, J=6.9Hz, 2H), 1.59- 1.56 (m, 1H), 1.32-1.28 (t, J=7.1, 14.2Hz, 3H), 0.86-0.85 (d, J=6.6Hz, 6H).

Step 2: 8-Isobutyl-7-methoxy-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of ethyl 8-isobutyl-7-methoxy-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylate (1.1 g, 0.0027 mol) in a mixture of THF (35 mL), water (10 mL), and methanol (5 mL) was added _LiOH.H₂O (332 mg, 0.0080 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was evaporated and acidified with 1.5 N HCl solution. The precipitated solid was filtered and dried to afford the desired compound (900 mg, 88%) as a creamy white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.33 (bs, 1H), 7.79-7.78 (dd, J＝1.4, 5.5Hz, 1H), 7.41-7.40 (dd, J＝1.4, 3.7Hz, 1H), 7.19-7.17 (dd, J＝3.8, 5.5Hz, 1H), 6.65 (s, 1H), 6.36 (s, 1H), 5.43 (s, 2H), 3.73 (s, 3H), 2.12-2.10 (d, J=7.0Hz, 2H), 1.59-1.56 (m, 1H), 0.71-0.70 (d, J=6.6Hz 6H). m/z: 385[M+H] ⁺ .

Step 3: 8-Isobutyl-7-methoxy-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide

To a solution of 8-isobutyl-7-methoxy-1-thiophen-2-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (900 mg, 0.0023 mol) in DCM (50 mL) was added N-tert-butylmethylamine (245 mg, 0.0028 mol), HATU (1.0 g, 0.0028 mol) and diisopropylethylamine (0.6 mL, 0.0038 mol) at room temperature and under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with sodium bicarbonate (10 mL, 10%) and extracted with DCM (2 x 50 mL). The organic layers were combined, washed with NaHCO ₃ solution (1 x 100 mL, 10% aqueous solution), brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo; the crude product was purified by column chromatography using petroleum ether and ethyl acetate (9:1) as eluent to afford the desired compound (850 mg, 90%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.76-7.75 (dd, J=1.4, 5.5Hz, 1H), 7.38-7.37 (dd, J=1.4, 3.7Hz, 1H), 7.18-7.15 (dd, J=3.7, 5.6Hz, 1H), 6.64 (s, 1H), 6.39 (s, 1H), 5 .30 (s, 2H), 3.73 (s, 3H), 3.12 (s, 3H), 3.12 (s, 3H), 2.12-2.10 (d, J = 7.0Hz, 2H), 1.60-1.56 (m, 1H), 1.41 (s, 9H) 0.71-0.70 (d, J = 6.6Hz 6H). m/z: 454 [M+H] ⁺ .

Process 5:

Example 23

(3,3-Dimethyl-morpholin-4-yl)-(8-isobutylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone (97)

Step 1: 8-Isobutylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (100.00 mg; 0.25 mmol; 1.00 eq.) in [1,4]dioxane (3.00 ml) was added 2-methyl-propane-1-thiol (33.22 mg; 0.37 mmol; 1.50 eq.), palladium acetate (2.76 mg; 0.01 mmol; 0.05 eq.), 4,5-bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene (14.21 mg; 0.02 mmol; 0.10 eq.), and potassium carbonate (101.81 mg; 0.74 mmol; 3.00 eq.). The reaction mixture was heated at 120°C for 8 days. The mixture was diluted with ethyl acetate and washed with brine. The organic layer was dried _{over Na2SO4} , filtered, and concentrated to give the product 8-isobutylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid as a yellow crude product.

Step 2: (3,3-Dimethyl-morpholin-4-yl)-(8-isobutylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone

To a suspension of 8-isobutylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (80.00 mg; 0.19 mmol; 1.00 eq.) in DCM (3.00 ml; 46.80 mmol; 243.67 eq.) was added 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.17 ml; 0.29 mmol; 1.50 eq.), 3,3-dimethylmorpholine (0.14 ml; 0.29 mmol; 1.50 eq.) and ethyl-diisopropylamine (0.10 ml; 0.58 mmol; 3.00 eq.). The reaction was stirred at room temperature for 1 hour. The mixture was concentrated and purified by flash chromatography to give the desired product (40 mg, 40%) as a white solid.

¹ H-NMR (DMSO-d6): δ 8.02-7.98 (s, 1H), 7.88-7.83 (m, 1H), 7.35 (dd, 1H), 6.72 (s, 1H), 6.60 (s, 1H), 5.37 (s, 2H), 3.94 (m, 2H), 3.80 (s, 3H), 3.72 (t, 2H), 3.42 (s, 2H), 2.34 (d, 2H), 1.60 (septet, 1H), 1.42 (s, 6H), 0.91 (d, 6H). m/z = 514 [M+H] ⁺ .

Example 24

8-Isopropylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (52)

Following the same procedure as in Example 23, 8-isopropylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide was prepared from 8-isopropylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (170 mg, 0.42 mmol) and n-tert-butylmethylamine (73.63 mg, 0.84 mmol, 2 eq.) in a 15% yield (29 mg) as a white solid. m/z = 472 [M+H] ⁺ , HPLC retention time = 6.45 min.

Example 25

7-Methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (79)

To a solution of 8-isopropylsulfanyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (24 mg, 0.05 mmol) in DCM (2 mL) was added 3-chloroperbenzoic acid (11.40 mg; 0.05 mmol; 1.00 eq.). The reaction mixture was stirred at room temperature for 2 hours. The mixture was purified by flash chromatography to give the desired product (8 mg, 32%) as a white solid. m/z = 504 [M+H] ⁺ , HPLC retention time = 4.40 min.

Example 26

(8-Cyclopropanesulfonyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone (106)

Step 1: 8-(Cyclopropylsulfonyl)-7-methoxy-1-(thien-3-yl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a suspension of 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (100.00 mg; 0.25 mmol; 1.00 eq.) in N,N-dimethylformamide (3.00 ml) were added sodium cyclopropanesulfinate (47.19 mg; 0.37 mmol; 1.50 eq.), copper iodide (23.38 mg; 0.12 mmol; 0.50 eq.) and N,N'-dimethylethane-1,2-diamine (0.04 ml; 0.37 mmol; 1.50 eq.). The reaction mixture was heated to 90°C for 18 hours. The mixture was filtered, concentrated, and lyophilized to give 8-(cyclopropylsulfonyl)-7-methoxy-1-(thiophen-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid as a blue solid.

Step 2: (8-Cyclopropanesulfonyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone

Following the same procedure as in Step 2 of Example 23 above, (8-cyclopropanesulfonyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone was prepared from 8-(cyclopropylsulfonyl)-7-methoxy-1-(thiophen-3-yl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylic acid (30.00 mg; 0.07 mmol; 1.00 eq.) and 3,3-dimethyl-morpholine (0.05 ml; 0.10 mmol; 1.50 eq.). The desired compound was obtained in 33% yield (12 mg) as a blue solid.

LCMS: m/z = 530 [M+H] ⁺ HPLC retention time = 3.24 min.

Example 27

(3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone (112)

Step 1: 7-Methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

Following the same procedure of Example 26 (step 1) above, 7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid was prepared from 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (100.00 mg; 0.25 mmol; 1.00 eq.) and sodium propane-2-sulfinate (47.94 mg; 0.37 mmol; 1.50 eq.) as a crude blue solid.

Step 2: (3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone

Following the same procedure as in Example 26 (Step 2) above, (3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl]-methanone was prepared from 7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-carboxylic acid (100.00 mg; 0.23 mmol; 1.00 eq.) and 3,3-dimethyl-morpholine (0.17 ml; 0.35 mmol; 1.50 eq.) in 8.2% yield (5 mg) as a blue solid. LCMS: m/z = 532 [M+H] ⁺ , HPLC retention time = 3.25 min.

Example 28

(3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-8-(2-methyl-propane-1-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone (127)

Step 1: 7-Methoxy-8-(2-methyl-propane-1-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

Following the same procedure of Example 26 (step 1) above, 7-methoxy-8-(2-methyl-propane-1-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid was prepared from 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (100.00 mg; 0.25 mmol; 1.00 eq.) and sodium 2-methyl-propane-1-sulfinate (53.10 mg; 0.37 mmol; 1.50 eq.) as a crude blue solid.

Step 2: (3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-8-(2-methyl-propane-1-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone

Following the same procedure of Example 26 (Step 2) above, (3,3-dimethyl-morpholin-4-yl)-[7-methoxy-8-(2-methyl-propane-1-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone was prepared from 7-methoxy-8-(2-methyl-propane-1-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (30.00 mg; 0.07 mmol; 1.00 eq.) and 3,3-dimethyl-morpholine (46.22 mg; 0.40 mmol; 6.00 eq.) in a yield of 13.7% (5 mg) as a white solid.

LCMS: m/z = 546 [M+H] ⁺ , HPLC retention time = 3.56 min.

Example 29

7-Methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-hydroxymethyl-oxetan-3-yl)-amide (187)

Following the same procedure of Example 27 above, 7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-hydroxymethyl-oxetane-3-yl)-amide ketone was prepared from 7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (70.00 mg; 0.16 mmol; 1.00 eq.) and (3-amino-oxetane-3-yl)-methanol (24.92 mg; 0.24 mmol; 1.50 eq.) in a yield of 25% (21 mg) as a white solid.

¹ H-NMR (DMSO-d6): δ8.77 (s, 1H), 8.02 (dd, 1H), 7.86 (dd, 1H), 7.34 (dd, 1H), 7.19 (s, 1H), 6.93 (s, 1H), 5. 64 (s, 2H), 5.14 (s, 1H), 4.67 (d, 2H), 4.51 (d, 2H), 3.90 (s, 3H), 3.68 (d, 2H), 3.45 (septet, 1H), 1.08 (d, 6H). m/z=520[M+H] ⁺ .

Example 30

7-Methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-methyl-oxetan-3-yl)-amide (188)

Following the same procedure of Example 27 above, 7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-methyl-oxetane-3-yl)-amide was prepared from 7-methoxy-8-(propane-2-sulfonyl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (70.00 mg; 0.16 mmol; 1.00 eq.) and 3-methyl-oxetane-3-ylamine hydrochloride (29.87 mg; 0.24 mmol; 1.50 eq.) in a yield of 34% (28 mg) as a white solid.

¹ H-NMR (DMSO-d6): δ8.95 (s, 1H), 8.03 (dd, 1H), 7.85 (dd, 1H), 7.34 (dd, 1H), 7.18 (s, 1H), 6.93 (s, 1 H), 5.64 (s, 2H), 4.71 (d, 2H), 4.31 (d, 2H), 3.90 (s, 3H), 3.45 (septet, 1H), 1.59 (s, 3H), 1.07 (d, 6H). m/z=504[M+H] ⁺ .

Example 31

(3,3-Dimethyl-morpholin-4-yl)-[8-(1-hydroxy-2-methyl-propyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone (131)

To a suspension of (3,3-dimethyl-morpholin-4-yl)-[7-methoxy-8-(2-methyl-propenyl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl]-methanone (70.00 mg; 0.15 mmol; 1.00 eq.) in THF (3.00 ml) was added methylsulfanylmethane and borane (16.63 mg; 0.22 mmol; 1.50 eq.). The reaction mixture was stirred at room temperature for 2 hours. NaOH (2N solution) was then slowly added. The reaction mixture was purified by reverse phase preparative HPLC (45-60% _CH₃CN in 0.1% aqueous _NH₄OH ) to afford the desired product (19 mg, 26%) as a white solid.

¹ H-NMR (DMSO-d6): δ7.86 (dd, 1H), 7.78 (dd, 1H), 7.28 (dd, 1H), 6.92 (s, 1H), 6.65 (s, 1H), 5.39 (d, 1H), 5.26 (d, 1H), 4.56 (d, 1H), 4.50 (t , 1H), 4.03-3.96(m, 1H), 3.93-3.85(m, 1H), 3.77–3.70(m, 5H), 3.42(s, 2H), 1.61(sextet, 1H), 1.42(d, 6H), 0.74(d, 3H), 0.68(d, 3H). m/z=498[M+H] ⁺ .

Example 32

(3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-1-thiophen-3-yl-8-(5-trimethylsilyl-5H-[1,2,4]triazol-3-yl)-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone (139)

To a solution of trimethylsilyldiazomethane (0.17 ml; 0.33 mmol; 3.00 eq.) in ethoxyethane (3.00 ml) at 0°C was added n-butyllithium (0.09 ml; 0.22 mmol; 2.00 eq.). The reaction mixture was stirred at 0°C for 30 minutes, followed by the addition of 3-(3,3-dimethylmorpholine-4-carbonyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-8-carbonitrile (50.00 mg; 0.11 mmol; 1.00 eq.), and the reaction mixture was stirred at 0°C for another 30 minutes. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The mixture was concentrated and purified by column chromatography to yield the desired product (18 mg, 29%) as a white solid.

¹ H-NMR (DMSO-d6): δ7.92 (dd, 1H), 7.72 (dd, 1H), 7.28 (dd, 1H), 6.79 (s, 1H), 6.73 (s, 1H), 5.43 (s , 2H), 3.96–3.92(m, 2H), 3.74–3.68(m, 5H), 3.42(s, 2H), 3.18(s, 1H), 1.43(s, 6H), 0.06(s, 9H). m/z=565[M+H] ⁺ .

Example 33

(3,3-Dimethyl-morpholin-4-yl)-[7-methoxy-1-thiophen-3-yl-8-(5H-[1,2,4]triazol-3-yl)-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl]-methanone (161)

To a solution of (3,3-dimethyl-morpholin-4-yl)-[7-methoxy-1-thiophen-3-yl-8-(5-trimethylsilyl-5H-[1,2,4]triazol-3-yl)-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl]-methanone (34.00 mg; 0.06 mmol; 1.00 eq.) in THF (3.00 ml) was added tetrabutylammonium fluoride (1 M in THF) (0.30 ml; 0.30 mmol; 5.00 eq.) and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated and purified by reverse phase preparative HPLC (35-40% CH ₃ CN in 0.1% aqueous NH ₄ OH) to give the desired product (36 mg, 82%) as a white solid.

¹ H-NMR (DMSO-d6) (tetrabutylammonium salt): δ 7.89 (dd, 1H), 7.79 (dd, 1H), 7.73 (s, 1H), 7.53 (s, 1H), 7.31 (dd, 1H), 6.70 (s, 1H), 5.31 (s, 2H), 3.96 (t, 2H), 3.83 (s, 3H), 3.73 (t, 2H), 3.43 (s, 2H), 3.20–3.13 (m, 9H), 1.64–1.52 (m, 9H), 1.43 (s, 6H), 1.33 (sextet, 9H), 0.94 (t, 12H). m/z = 493 [M+H] ⁺ .

Example 34

(8-Aminomethyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone (166)

Step 1: Ethyl 8-cyano-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate

To a solution of ethyl 8-bromo-7-methoxy-1-(3-thienyl)-1,4-dihydrochromeno[4,3-c]pyrazole-3-carboxylate (2 g, 0.0459 mol) in NMP (50 mL) in a sealed tube was added CuI (90 mg, 0.5 mmol) followed by CuCN (825 mg, 9.1 mmol). The reaction mixture was heated to 160°C for 16 hours. The reaction mixture was filtered through celite, and the filtrate was concentrated. The crude product was purified by column chromatography using dichloromethane/methanol (9:1) as the eluent. The product was triturated with acetonitrile and filtered to yield the desired compound (0.7 g, 83%) as a white solid.

^1H NMR (400MHz, DMSO) δ8.05-8.04 (dd, J=1.4, 3.1Hz, 1H), 7.90-7.88 (dd, J=3.2, 5.1Hz, 1H), 7.37-7.3 5 (dd, J=1.4, 5.1Hz, 1H), 5.63 (s, 1H), 4.33-4.28 (m, 2H), 3.89 (s, 1H), 1.32-1.28 (t, J=12.9Hz, 3H). m/z=382[M+H] ⁺ .

Step 2: 8-cyano-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of 8-cyano-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ethyl ester (300.00 mg; 0.79 mmol; 1.00 eq.) in methanol (6.00 ml; 177.52 mmol; 225.69 eq.) was added potassium hydroxide (66.20 mg; 1.18 mmol; 1.50 eq.) and water (0.60 ml). The reaction mixture was heated to 50°C for 2 hours. The reaction was concentrated and lyophilized to give the crude product as a gray solid.

Step 3: 3-(3,3-Dimethyl-morpholine-4-carbonyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-8-carbonitrile (86)

To a suspension of 8-cyano-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (270.00 mg; 0.76 mmol; 1.00 eq.) in DCM (6.00 ml; 93.60 mmol; 122.50 eq.) was added 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (1.35 ml; 2.29 mmol; 3.00 eq.), 3,3-dimethylmorpholine (176.01 mg; 1.53 mmol; 2.00 eq.) and ethyl-diisopropylamine (0.38 ml; 2.29 mmol; 3.00 eq.). The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated and purified by flash chromatography to give the desired product (309 mg, 90%) as a white solid. LCMS: m/z = 451 [M+H] ⁺ .

Step 4: (8-Aminomethyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone

3-(3,3-Dimethyl-morpholine-4-carbonyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-8-carbonitrile (50.00 mg; 0.11 mmol; 1.00 eq.) was dissolved in ammonia in methanol (10.00 ml; 20.00 mmol; 180.20 eq.). The reaction mixture was passed through an H-cube under full hydrogen pressure at 70°C using a Raney nickel tube. The mixture was concentrated and purified by reverse-phase preparative HPLC (35-45% _CH₃CN in 0.1% aqueous _NH₄OH ) to afford the desired product (7 mg, 14%) as a white solid. LCMS: m/z = 455 [M+H] ^⁺ , HPLC retention time = 2.71 min.

Example 35

N-[3-(3,3-Dimethyl-morpholine-4-carbonyl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-8-ylmethyl]-acetamide (175)

To a suspension of (8-aminomethyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone (20.00 mg; 0.04 mmol; 1.00 eq.) in DCM (2.00 ml; 31.20 mmol; 709.11 eq.) was added 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.04 ml; 0.07 mmol; 1.50 eq.), acetic acid (3.96 mg; 0.07 mmol; 1.50 eq.) and ethyl-diisopropylamine (0.02 ml; 0.13 mmol; 3.00 eq.). The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated and purified by reverse phase preparative HPLC (32-38% _CH3CN in 0.1% aqueous _NH4OH ) to give the desired product (4 mg, 18%) as a white solid.

¹ H-NMR (DMSO-d6): δ7.91-7.76 (m, 2H), 7.28-7.22 (m, 1H), 6.74-6.61 (m, 2H), 5.37-5.26 (m, 2H), 4.14 (s, 0.5H), 3.96(d, 3.5H), 3.84-3.69(m, 5H), 3.42(s, 2H), 2.05(s, 0.5H), 1.91(s, 0.5H), 1.86–1.76(m, 3H), 1.42(s, 6H). m/z=497[M+H] ⁺ .

Example 36

8-Cyano-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (12)

To a solution of 8-bromo-7-methoxy-1-thiophene-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (210 mg, 0.44 mmol) in NMP (5 mL) was added CuCN (43.5 mg, 0.48 mmol, 1.1 eq.) and CuI (8.4 mg, 0.044 mmol 0.1 eq.). The reaction mixture was heated in microwaves at 170° C. for 70 minutes. The mixture was purified by reverse phase preparative HPLC to give the desired product (10 mg, 5%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ7.54 (ddd, J=4.7, 4.1, 2.4Hz, 2H), 7.19 (dd, J=5.0, 1.5Hz, 1H), 6.97 (s, 1H), 6.62 (s, 1H), 5.58 (s, 2H), 3.92 (s, 3H), 3.28 (s, 3H), 1.53 (s, 9H). m/z=423[M+H] ⁺ .

Example 37

7-Methoxy-8-(1-methyl-1H-pyrrol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (2-hydroxy-1-hydroxymethyl-1-methyl-ethyl)-amide (179)

To 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-methyl-oxetan-3-yl)-amide (1.00 g; 2.10 mmol; 1.00 eq.) was added 1-methyl-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrrole (652.08 mg; 3.15 mmol). [0148] To the reaction mixture were added 1,2-dicyclohexyl-2-nitropropane (2',6'-dimethoxy-2-phenyl-1-yl)-phosphane (86.18 mg; 0.21 mmol; 0.10 eq.), potassium carbonate (870.42 mg; 6.30 mmol; 3.00 eq.), dioxane (10.00 ml), and water (1.00 ml). The reaction mixture was heated at 120°C for 24 hours. The mixture was concentrated, filtered, and purified by reverse-phase preparative HPLC (35-45% _CH₃CN in 0.1% aqueous _NH₄OH ) to afford the hydrolyzed product (14 mg, 1.3%) as a white solid.

¹ H-NMR (DMSO-d6): δ8.06 (s, 1H), 7.90 (s, 1H), 7.39 (s, 1H), 7.31 (s, 1H), 7.00 (s, 1H), 6.79 (s, 1H), 6.71 (s, 1H), 6. 63 (s, 1H), 5.77 (s, 1H), 5.49 (s, 2H), 4.95 (s, 2H), 3.83 (s, 3H), 3.65-3.56 (m, 5H), 3.53-3.45 (m, 2H), 1.30 (s, 3H). m/z=495[M+H] ⁺ .

Example 38

7-Methoxy-8-(1-methyl-1H-pyrazol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (2-hydroxy-1-hydroxymethyl-1-methyl-ethyl)-amide (185)

According to the same procedure of Example 37, 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (3-methyl-oxetan-3-yl)-amide (1.00 g; 2.10 mmol; 1.00 eq.) and 1-methyl-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolane-2 -yl)-1H-pyrazole (655.19 mg; 3.15 mmol; 1.50 eq.) was used to prepare 7-methoxy-8-(1-methyl-1H-pyrazol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (2-hydroxy-1-hydroxymethyl-1-methyl-ethyl)-amide in a yield of 2% (20 mg) as a white solid.

¹ H-NMR (DMSO-d6): δ8.01 (s, 1H), 7.85 (d, 1H), 7.60 (s, 1H), 7.41 (s, 1H), 7.32 (s, 2H), 6.76 (s, 1H), 6.49 (s, 1 H), 5.53 (s, 2H), 4.95 (s, 2H), 3.84 (s, 3H), 3.78 (s, 3H), 3.66-3.57 (m, 2H), 3.53-3.45 (m, 2H), 1.30 (s, 3H). m/z=496[M+H] ⁺ .

Example 39

7-Methoxy-8-(tetrahydro-furan-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (194)

To a suspension of 8-(2,5-dihydro-furan-3-yl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (30.00 mg; 0.06 mmol; 1.00 eq.) in acetic acid (3.00 ml) was added palladium on carbon (0.02 ml; 0.32 mmol; 5.00 eq.). The flask was capped with a rubber stopper topped with a hydrogen balloon. The reaction mixture was stirred at room temperature for 18 hours. Triethylamine was added, and the mixture was filtered through celite. The mixture was purified by reverse-phase preparative HPLC (55-63% _CH₃CN in 0.1% aqueous _NH₄OH ) to afford the desired product (11 mg, 37%) as a white solid.

¹ H-NMR (DMSO-d6): δ7.95 (dd, 1H), 7.84 (dd, 1H), 7.32 (dd, 1H), 6.69 (s, 1H), 6.60 (s, 1H), 5.34 (d, 2H), 3.85 (t, 1H ), 3.79(s, 3H), 3.71–3.57(m, 2H), 3.43(quintet, 1H), 3.21–3.15(m, 4H), 2.10–2.00(m, 1H), 1.56-1.40(m, 10H). m/z=468[M+H] ⁺ .

Example 40

7-Methoxy-8-(tetrahydro-furan-2-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (201)

Following the same procedure of Example 39 above, 7-methoxy-8-(tetrahydro-furan-2-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide was prepared from 8-(4,5-dihydro-furan-2-yl)-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (65.00 mg; 0.14 mmol; 1.00 eq.) in a yield of 9% (6 mg) as a white solid.

¹ H-NMR (DMSO-d6): δ7.93 (dd, 1H), 7.82 (dd, 1H), 7.30 (dd, 1H), 6.75 (s, 1H), 6.66 (s, 1H), 5.33 (q, 2H), 4.90 (dd, 1H), 3.7 7(s, 3H), 3.65-3.58(m, 2H), 3.17(s, 3H), 2.18-2.07(m, 1H), 1.85-1.74(m, 1H), 1.70-1.59(m, 1H), 1.50-1.40(m, 10H). m/z=468[M+H] ⁺ .

Example 41

7-Methoxy-8-(1-methyl-1H-pyrazol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester (180)

Step 1: To a solution of 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (500.00 mg; 1.23 mmol; 1.00 eq.) in DCM (10.00 ml; 156.01 mmol; 127.06 eq.) was added (1S,3S)-3-amino-cyclopentanol hydrochloride (253.43 mg; 1.84 mmol; 1.50 eq.), [(benzotriazol-1-yloxy)-dimethylamino-methylene]-dimethyl-ammonium tetrafluoroborate (788.43 mg; 2.46 mmol; 2.00 eq.) and ethyl-diisopropylamine (0.61 ml; 3.68 mmol; 3.00 eq.). The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated and purified by flash chromatography to give 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ((1S,3S)-3-hydroxy-cyclopentyl)-amide and 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester (247 mg, 41% in total) as white solids.

Step 2: To a mixture of 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ((1S,3S)-3-hydroxy-cyclopentyl)-amide and 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester (120.00 mg; 0.24 mmol; 1.00 eq.) was added 1-methyl-3-(4,4,5,5-tetramethyl-[ The reaction mixture was heated at 140° C. for 18 hours. The mixture was concentrated and a portion of the mixture was purified by flash chromatography (KPNH, 80-100% ethyl acetate/hexanes, 0-20% methanol/ethyl acetate) to give 7-methoxy-8-(1-methyl-1H-pyrazol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid ((1S,3S)-3-hydroxy-cyclopentyl)-amide (78 mg, 65%) as a white solid. The remaining crude material was purified by reverse phase preparative HPLC (35-45% CH ₃ CN in 0.1% aqueous NH ₄ OH) to afford 7-methoxy-8-(1-methyl-1H-pyrazol-3-yl)-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester (5 mg, 4.2%) as a white solid.

¹ H-NMR (DMSO-d6): δ7.94(s, 1H), 7.82(s, 1H), 7.60(s, 1H), 7.50(s, 1H), 7.33(s, 1H), 6.77(s, 1H ), 6.50 (s, 1H), 5.38 (s, 2H), 4.96 (s, 1H), 3.85 (s, 3H), 3.78 (s, 3H), 3.63 (s, 1H), 1.32 (d, 12H). m/z=492[M+H] ⁺ .

Example 42

7-Methoxy-8-(1-methyl-1H-pyrrol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester (181)

According to the same procedure of Example 41 above, 7-methoxy-8-(1-methyl-1H-pyrrol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester was prepared from 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (1S,3S)-3-amino-cyclopentyl ester in a yield of 3% (3 mg) as a white solid.

¹ H-NMR (DMSO-d6): δ8.01(s, 1H), 7.88(s, 1H), 7.38(s, 1H), 7.01(s, 1H), 6.88(s, 1H), 6.72(s, 1H), 6.64(s , 1H), 5.81(s, 1H), 5.34(s, 2H), 4.98(s, 1H), 4.09(s, 5H), 3.84(s, 3H), 3.59(s, 4H), 1.48–1.14(m, 12H). m/z=491[M+H] ⁺ .

Example 43

7-Methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (133)

To 8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (70.00 mg; 0.15 mmol; 1.00 eq.) was added boric acid, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ii) complex with dichloromethane (1:1) (24.00 mg; 0.03 mmol; 0.20 eq.), dioxane (2.00 ml; 23.47 mmol; 159.74 eq.), cesium carbonate (143.63 mg; 0.44 mmol; 3.00 eq.), and water (0.20 ml). The reaction mixture was heated at 120° C. for 18 hours. The reaction mixture was purified by reverse phase preparative HPLC (45-55% _CH3CN in 0.1% aqueous _NH4OH ) to afford 7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid tert-butyl-methyl-amide (30 mg, 51%) as a white solid.

¹ H-NMR (DMSO-d6): δ7.94 (dd, 1H), 7.81 (dd, 1H), 7.32 (dd, 1H), 6.67 (d, 1H), 6. 62 (d, 1H), 6.47 (dd, 1H), 5.33 (s, 2H), 3.73 (s, 3H), 3.17 (s, 3H), 1.44 (s, 9H). m/z=398[M+H] ⁺ .

Example 44

(2-Methoxymethyl-2-methyl-pyrrolidin-1-yl)-(7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone (183)

Following the same procedure of Example 43 above, (2-methoxymethyl-2-methyl-pyrrolidin-1-yl)-(7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone was prepared from (2-hydroxymethyl-2-methyl-pyrrolidin-1-yl)-(7-methoxy-1-thiophene-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone with a yield of 50% (13 mg) as a white solid.

¹ H-NMR (DMSO-d6): δ7.95(s, 1H), 7.82(s, 1H), 7.32(s, 1H), 6.69–6.60(m, 2H), 6.49-6.44(m, 1H), 5.43(s, 2H), 4.05(s, 1H) , 3.90-3.79(m, 2H), 3.73(s, 3H), 3.59-3.52(m, 1H), 2.16–2.07(m, 1H), 1.88-1.72(m, 2H), 1.68-1.58(m, 1H), 1.42(s, 3H). m/z=443[M+H] ⁺ .

Example 45

(3,3-Dimethyl-morpholin-4-yl)-(7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone (128)

Following the same procedure of Example 43 above, (3,3-Dimethyl-morpholin-4-yl)-(7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-methanone was prepared from (8-bromo-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazol-3-yl)-(3,3-dimethyl-morpholin-4-yl)-methanone with a yield of 46% (13 mg) as a white solid.

LCMS: m/z = 427 [M+H] ⁺ , HPLC retention time = 3.65 min.

Example 46

7-Methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3,8-dicarboxylic acid 8-amide 3-(tert-butyl-methyl-amide)(50)

Step 1: 8-Carbamoyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid

To a solution of 8-cyano-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (45.00 mg; 0.13 mmol; 1.00 eq.) (Example 34) in DMSO (4 mL) was added _{H₂O₂ (} 0.12 ml; 1.27 mmol; 10.00 eq.) and 2.0 M aqueous NaOH (0.64 ml; 1.27 mmol; 10.00 eq.). The reaction mixture was stirred at room temperature for 2 hours. The mixture was purified by reverse phase HPLC to give the desired product (17 mg, 35%) as a white solid.

Step 2: 7-Methoxy-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3,8-dicarboxylic acid 8-amide 3-(tert-butyl-methyl-amide)

To a solution of 8-carbamoyl-7-methoxy-1-thiophen-3-yl-1,4-dihydro-chromeno[4,3-c]pyrazole-3-carboxylic acid (17 mg, 0.05 mmol) in DCM (1.00 ml; 46.80 mmol; 1022.39 eq.) was added DIPEA (0.02 ml; 0.09 mmol; 2.00 eq.), O-benzotriazole-n,n,n',n'-tetramethyluronium tetrafluoroborate (29.40 mg; 0.09 mmol; 2.00 eq.), and N-tert-butylmethylamine (0.01 ml; 0.09 mmol; 2.00 eq.). The reaction mixture was stirred at room temperature for 30 minutes. The mixture was concentrated and purified by reverse phase preparative HPLC to give the desired product (2.5 mg, 11%) as a white solid.

¹ H NMR (400MHz, MeOD) δ7.88 (s, 1H), 7.71 (s, 1H), 7.69–7.56 (m, 1H), 7.26 (d, J＝ 4.9Hz, 1H), 6.79 (s, 1H), 5.46 (s, 2H), 3.98 (s, 3H), 3.23 (s, 3H), 1.54 (s, 9H). m/z=441[M+H] ⁺ .

Example 47

The following compounds were prepared according to the method disclosed in Example 1:

Example 48

N-(1,3-Dihydroxy-2-methylprop-2-yl)-7-methoxy-N-methyl-8-(1-methyl-1H-pyrazol-3-yl)-1-(thien-3-yl)-1,4-dihydrobenzopyrano[4,3-c]pyrazole-3-carboxamide (217)

Compound 116 (MSC2501240) (30.00 mg; 0.06 mmol; 1.00 eq.) was dissolved in a solution of hydrochloric acid in water (2.00 ml). The reaction mixture was stirred at room temperature overnight. The mixture was subjected to preparative HPLC (32-38% _CH₃CN in 0.1% aqueous _NH₄OH ) to afford the desired product, 7-methoxy-8-(1-methyl-1H-pyrazol-3-yl)-1-thiophen-3-yl-1,4-dihydro-benzopyrano[4,3-c]pyrazole-3-carboxylic acid (2-hydroxy-1-hydroxymethyl-1-methyl-ethyl)-methyl-amide (19.00 mg; 0.04 mmol) (61%), as a white solid. LCMS: m/z = 513 [M+H] ^⁺ , HPLC retention time: 2.67 min.

Example 49

_EC50 of cyclic AMP production in CHO FSHR cells in the presence of _EC20 FSH (Assay A)

2500 Cho-FSHR-LUC-1-1-43 cells were plated per well in 5 μl of phenol red-free DMEM/F12 + 1% FBS. The cells were plated into 384-well white low-volume culture plates (Greiner 784075) using a dispenser. 100 μl (2X) of EC ₂₀ FSH/IBMX in DMEM/F12 and 0.1% BSA was added to 2 μl of test compound (compound diluted 1:50) plated in the 384-well culture plates using a dispenser. The final concentration of FSH was 0.265 pM, and the final concentration of IBMX was 200 μM. The compound plate layout was as follows: Column 1: 2 μl DMSO; Column 2: 2 μl DMSO; Columns 3-12 and 13-24: 2 μl of test compound diluted 1:4 with 100% DMSO or 2 μl of FSH diluted 1:4 with DMEM/F12 + 0.1% BSA. The starting concentration of FSH was 50 nM (final concentration 0.5 nM). In addition, column 23 contained 2 μl of _EC100 FSH reference (100X) (diluted in DMEM/F12 + 0.1% BSA) to a final concentration of 0.5 nM, and column 24 contained 2 μl of 1 mM AS707664/2 reference compound. A 2.5 μl mixture of compound and _EC20 FSH was transferred to the cell culture plate (5 μl of cell culture medium was added to the 1:2 dilution). The plate was incubated at 37°C for 1 hour. 10 μl of mixed HTRF reagent (CisBio #62AM4PEC) was added to each well and incubated at room temperature for 1 hour. The plate was read using the Envision cAMP HTRF-Low Volume 384-well protocol. The reading was calculated as the fluorescence ratio (665 nm/620 nm). Values were expressed as a percentage (%), indicating the percent effect (response) of a given agonist concentration relative to the maximum response of an FSH standard. The results are listed in the table below.

Example 50

Rat granulosa cells EC ₅₀ FSH (Test B)

This experiment was conducted according to the following literature: Yanofsky et al. (2006), Allosteric activation of the follicle-stimulating hormone (FSH) receptor by selective, nonpeptide agonists. JBC 281(19): 13226-13233, which is incorporated herein by reference. The results are listed in the table below.

Understand the data using the following definitions:

Example 51

pharmaceutical preparations

(A) Injection vial: A solution of 100 g of the compound of the present invention as the active ingredient and 5 g of sodium dihydrogen phosphate in 3 L of redistilled water is adjusted to pH 6.5 with 2N hydrochloric acid, sterile filtered, transferred to an injection vial, lyophilized under aseptic conditions, and sealed under aseptic conditions. Each injection vial contains 5 mg of the active ingredient.

(B) Suppositories: 20 g of the compound of the present invention as the active ingredient is mixed with 100 g of soybean lecithin and 1400 g of cocoa butter, poured into a mold, and cooled. Each suppository contains 20 mg of the active ingredient.

(C) Solution Formulation: Prepare a solution consisting of 1 g of the active ingredient compound of the present invention, 9.38 g _{of NaH₂PO₄} · _2H₂O , 28.48 g _{of Na₂HPO₄} · _12H₂O , and 0.1 g of benzalkonium chloride in 940 mL of redistilled water. Adjust the pH of the solution to 6.8, make up to 1 L, and sterilize by radiation. Use the solution as eye drops.

(D) Ointment: 500 mg of the compound of the present invention as an active ingredient is mixed with 99.5 g of vaseline under sterile conditions.

(E) Tablets: 1 kg of the compound of the present invention as an active ingredient, 4 kg of lactose, 1.2 kg of potato flour, 0.2 kg of talc and 0.1 kg of magnesium stearate are compressed into tablets according to a conventional method so that each tablet contains 10 mg of the active ingredient.

(F) Coated tablets: Tablets are compressed similarly to Example E and then coated with sucrose, potato flour, talc, tragacanth and dye according to conventional methods.

(G) Capsules: 2 kg of the compound of the present invention as an active ingredient is introduced into hard capsules according to a conventional method so that each capsule contains 20 mg of the active ingredient.

(H) Ampoules: A solution of 1 kg of the compound of the present invention as the active ingredient in 60 L of redistilled water is sterile filtered, transferred into ampoules, lyophilized under aseptic conditions, and sealed under aseptic conditions. Each ampoule contains 10 mg of the active ingredient.

(I) Inhalation spray: Dissolve 14 g of the active ingredient of the compound of the present invention in 10 L of isotonic sodium chloride solution. Transfer the solution to a commercially available spray container with a pump mechanism. The solution can be sprayed into the mouth or nose. One spray (approximately 0.1 ml) is equivalent to a dose of approximately 0.14 mg.

Although many embodiments of the present invention are described herein, it is apparent that the basic embodiments can be modified using the compounds and methods of the present invention to provide other embodiments. It should be understood, therefore, that the scope of the invention is to be limited by the appended claims rather than by the specific embodiments provided by way of example.

Claims (5)

1. A compound selected from the following compounds, or pharmaceutically acceptable salts thereof: 2. A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable adjuvant, carrier, or mediator. 3. Use of the compound of claim 1 or a physiologically acceptable salt thereof in the preparation of a medicament for regulating the activity of FSHR or its mutants in a patient or biological sample. 4. Use of the compound of claim 1 or a physiologically acceptable salt thereof in the preparation of a medicament for treating fertility disorders in a subject. 5. Use of the compound of claim 1 or a physiologically acceptable salt thereof in the preparation of a medicament for the preventive or therapeutic treatment of diseases mediated by FSHR.